PMID- 18506356 TI - Topical therapy for the management of childhood psoriasis: part I. AB - Psoriasis represents a potentially life-altering disease that can profoundly impact physical, emotional and social functioning, and overall quality of life. The majority of cases are mild and managed adequately with topical medications. A minor subset of children present with severe, rapidly evolving disease that requires systemic therapy. The choice of treatment in children, as in adults, is determined by disease acuity, morphology, distribution, severity and the presence of comorbidities such as psoriatic arthropathy. Practical considerations such as ease of use, patient acceptability, accessibility, risk to benefit ratio, cost and individual perceptions of disease and quality of life are factored into treatment decisions. Part I of this 2-part series will focus on topical agents, their varying degrees of effectiveness, potential side-effects and applications in clinical practice. PMID- 18506357 TI - Advances in pemphigus therapy. AB - The pemphigus variants represent a group of potentially life-threatening autoimmune mucocutaneous blistering diseases. Though systemic corticosteroids have dramatically reduced the rate of disease mortality, current therapeutic options are limited by their toxicity profiles. Advancements in our understanding of the molecular mechanisms involved in the pathogenesis of pemphigus have translated into the development of novel therapies. However, few treatments have been subject to randomized controlled trials to firmly establish therapeutic efficacy. Herein, we focus on the new and emerging therapies in the management of pemphigus. PMID- 18506359 TI - A dipeptide YY derived from royal jelly proteins inhibits renin activity. AB - Renin is the rate limiting enzyme in the renin-angiotensin (RA) system that regulates blood pressure and electrolyte balance. In this study, we investigated the renin inhibitory effect of a royal jelly (RJ)-derived peptide. A dipeptide YY was isolated from the digested fraction of RJ proteins by proteases and was found to inhibit human renin activity. The inhibition constant (Ki) of YY was estimated to be 10 microM when the Km was 0.16 microM using sheep angiotensinogen as the substrate. The peptide was observed to lower blood pressure in spontaneously hypertensive rats. PMID- 18506358 TI - Aldose reductase inhibitor fidarestat counteracts diabetes-associated cataract formation, retinal oxidative-nitrosative stress, glial activation, and apoptosis. AB - This study was aimed at evaluating the potent and specific aldose reductase inhibitor fidarestat, on diabetes-associated cataract formation, and retinal oxidative-nitrosative stress, glial activation, and apoptosis. Control and streptozotocin-diabetic rats were treated with or without fidarestat (16 mg kg( 1)d(-1)) for 10 weeks after an initial 2-week period without treatment. Lens changes were evaluated by indirect ophthalmoscopy and portable slit lamp. Nitrotyrosine, poly(ADP-ribose), and glial fibrillary acidic protein expression were assessed by immunohistochemistry. The rate of apoptosis was quantified in flat-mounted retinas by TUNEL assay with immunoperoxidase staining. To dissect the effects of high glucose exposure in retinal microvascular cells, primary bovine retinal pericytes and endothelial cells were cultured in 5 or 30 mM glucose, with or without fidarestat (10 microM) for 3-14 days. Apoptosis was assessed by TUNEL assay, nitrotyrosine and poly(ADP-ribose) by immunocytochemistry, and Bax and Bcl-2 expression by Western blot analyses. Fidarestat treatment prevented diabetic cataract formation and counteracted retinal nitrosative stress, and poly(ADP-ribose) polymerase activation, as well as glial activation. The number of TUNEL-positive nuclei (mean +/- SEM) was increased approximately 4-fold in diabetic rats vs. controls (207+/-33 vs. 49+/ 4, p<0.01), and this increase was partially prevented by fidarestat (106+/-34, p<0.05 vs. untreated diabetic group). The apoptotic cell number increased with the prolongation of exposure of both pericytes and endothelial cells to high glucose levels. Fidarestat counteracted nitrotyrosine and poly(ADP-ribose) accumulation and apoptosis in both cell types. Antiapoptotic effect of fidarestat in high glucose-exposed retinal pericytes was not associated with the inhibition of Bax or increase in Bcl-2 expression. In conclusion, the findings, i) support an important role for aldose reductase in diabetes-associated cataract formation, and retinal oxidative-nitrosative stress, glial activation, and apoptosis, and ii) provide a rationale for the development of aldose reductase inhibitors, and, in particular, fidarestat, for the prevention and treatment of diabetic ocular complications. PMID- 18506360 TI - Neuromedin-U stimulates enucleation-induced adrenocortical regeneration in the rat. AB - Neuromedin-U (NMU) is a brain-gut peptide, which has been previously found to stimulate hypothalamic-pituitary-adrenal axis in the rat. Enucleation-induced adrenal regeneration in rats with contralateral adrenalectomy is a well established model of adrenal growth, that not only depends on the compensatory ACTH hypersecretion, but is also modulated by several regulatory peptides. Hence, we investigated whether NMU may be included in this group of bioactive molecules. Reverse transcription-polymerase chain reaction and immunocytochemistry showed that regenerating rat adrenocortical cells at days 5 and 8 after surgery express the NMU receptor NMUR1 as mRNA and protein. NMU8 administration to rats bearing regenerating adrenals markedly raised the plasma concentration of corticosterone and notably enhanced proliferative activity of adrenocortical cells. ACTH blood level was unchanged at day 5 and significantly decreased at day 8. The conclusion is drawn that NMU stimulates regeneration of rat adrenal cortex, via a mechanism independent of pituitary ACTH and involving the activation of NMUR1 located on adrenocortical cells. PMID- 18506362 TI - Expression profiling of macrophages from subjects with atherosclerosis to identify novel susceptibility genes. AB - Although a number of environmental risk factors for atherosclerosis have been identified, heredity seems to be a significant independent risk factor. The aim of our study was to identify novel susceptibility genes for atherosclerosis. The screening process consisted of three steps. First, expression profiles of macrophages from subjects with atherosclerosis were compared to macrophages from control subjects. Secondly, the subjects were genotyped for promoter region polymorphisms in genes with altered gene expression. Thirdly, a population of subjects with coronary heart disease and control subjects were genotyped to test for an association with identified polymorphisms that affected gene expression. Twenty-seven genes were differentially expressed in both macrophages and foam cells from subjects with atherosclerosis. Three of these genes, IRS2, CD86 and SLC11A1 were selected for further analysis. Foam cells from subjects homozygous for the C allele at the -765C-->T SNP located in the promoter region of IRS2 had increased gene expression compared to foam cells from subjects with the nonCC genotype. Also, macrophages and foam cells from subjects homozygous for allele 2 at a repeat element in the promoter region of SLC11A1 had increased gene expression compared to macrophages and foam cells from subjects with the non22 genotype. Genotyping of 512 pairs of subjects with coronary heart disease (CHD) and matched controls revealed that subjects homozygous for C of the IRS2 SNP had an increased risk for CHD; odds ratio 1.43, p=0.010. Immunohistochemical staining of human carotid plaques showed that IRS2 expression was localised to macrophages and endothelial cells in vivo. Our method provides a reliable approach for identifying susceptibility genes for atherosclerosis, and we conclude that elevated IRS2 gene expression in macrophages may be associated with an increased risk of CHD. PMID- 18506361 TI - Induction of tumor cell apoptosis by a novel class of N-thiolated beta-lactam antibiotics with structural modifications at N1 and C3 of the lactam ring. AB - The investigation of novel anti-tumor agents that preferentially select for malignant cells with a tolerable toxicity level has been the focus of anti-cancer drug discovery. Our laboratories have previously reported that certain N alkylthiolated beta-lactams had DNA-damaging and apoptosis-inducing activity in various tumor lines but not in nontransformed cells. In the current study, we further delineated the effects of substitutions at C3 or N1 of the lactam ring for cell death-inducing capability with close attention paid to a discernible structure-activity relationship (SAR). We found that two beta-lactam analogs (JG 5 and JG-19), both containing a branched-chain moiety at C3 of the lactam ring, exhibit potent apoptosis-inducing activity. Additionally, JG-5 exhibited superior in vitro biological activity over JG-19 owing to structural modifications made to substituents at the N1 and C3 positions of the lactam ring. Furthermore, the branched beta-lactams were able to inhibit growth of mice bearing breast cancer xenografts, associated with induction of DNA damage and apoptosis in tumor tissues. Our results strongly warrant further investigation into these novel beta lactams as potential anti-cancer therapeutics. PMID- 18506363 TI - Thirty-two new cases with small supernumerary marker chromosomes detected in connection with fertility problems: detailed molecular cytogenetic characterization and review of the literature. AB - Thirty-two patients with fertility problems were identified as carriers of small supernumerary marker chromosomes (sSMC). Molecular cytogenetic techniques were used to characterize their chromosomal origin. Together with the other cases available in the literature 111 sSMC cases have now been detected in connection with fertility problems in otherwise clinically healthy persons and characterized for their genetic content. According to this study, in 60% of the cases the sSMC originated from chromosomes 14 or 15. Euchromatic imbalances were caused by the sSMC presence in 30% of the cases. Notably, in 53% of infertile sSMC carriers, the sSMC was parentally transmitted. As we found indications of an as yet unknown mechanism for the elimination of sSMC from the human gene pool, sSMC could also play a role in elucidating the process of chromosome gain and loss during evolution. Nonetheless, further detailed molecular analysis will be necessary in the future to characterize the mechanisms and genetic basis for this phenomenon. PMID- 18506364 TI - Evaluation of the common variants of the ABCA4 gene in families with Stargardt disease and autosomal recessive retinitis pigmentosa. AB - Stargardt disease (STGD) is one of the most common autosomal recessive retinal dystrophies with an estimated incidence of one in 10,000. It affects the central retina (macula). Retinitis pigmentosa (RP) comprises a large and exceptionally heterogeneous group of hereditary disorders of the retina. It is caused by the loss of photoreceptors. The condition is a degenerative disorder characterized by retinal pigment deposits and has an estimated incidence of one in 4,000. Although, to date, 45 known loci have been identified, none of them independently account for a substantial portion of RP. Recently, the photoreceptor cell specific ATP-binding cassette transporter (ABCA4) gene was found to be mutated in patients with STGD as well as autosomal recessive RP. In order to further understand the contribution of this gene to the susceptibility to STGD and RP, we analyzed three unrelated STGD families and one autosomal recessive RP family specifically for the more common variants (A1038V, G1961E, 2588G-->C, R943Q or 2828G-->A) in the ABCA4 gene. Our analyses employing standard techniques such as polymerase chain reaction, restriction fragment length polymorphism, and direct DNA sequencing of amplified products were able to identify one common variant (R943Q) in all three STGD families but not in the RP family. All three affected STGD individuals, however, were heterozygous for this variation, and this alteration did not segregate with the disease and was also present in the normal controls. Similar analysis of other common variants revealed no pathogenic mutations in the STGD and RP families. It is likely that the variant identified in this study represents a rare polymorphism (non-pathogenic). Although, at present we cannot eliminate the possibility of this gene as a candidate gene, future extensive studies on this as well as other candidate genes may uncover the susceptibility gene for these recessive forms of the disorders in these families. PMID- 18506365 TI - Pyrogallol as a glutathione depletor induces apoptosis in HeLa cells. AB - Pyrogallol, a polyphenol, is known to be a superoxide anion (O2(.-)) generator. We investigated the involvement of glutathione (GSH) and reactive oxygen species (ROS) in pyrogallol-induced HeLa cell death. We measured the changes of ROS levels, GSH levels, sub-G1 cells, annexin V/PI staining cells and mitochondria membrane potential (DeltaPsi m) in HeLa cells treated with pyrogallol and/or ROS scavenger. The intracellular ROS levels were decreased or increased depending on the concentration of pyrogallol. The level of O2(.-) was significantly increased and superoxide dismutase (SOD) activity was down-regulated by pyrogallol. Pyrogallol reduced intracellular GSH content in HeLa cells. The ROS scavengers, Tempol, Tiron, Trimetazidine and N-acetylcysteine (NAC), did not down-regulate the production of O2(.-). However, treatment with NAC showed the recovery of GSH depletion and significantly rescued cells from pyrogallol-induced apoptosis. In addition, the recovery of GSH depletion by SOD and catalase was accompanied by the decrease of apoptosis levels. Furthermore, NAC and SOD significantly inhibited CMF-negative (GSH-depleted) and PI-positive cells induced by pyrogallol. Taken together, pyrogallol potently increased intracellular O2(.-) levels and decreased GSH content in HeLa cells, and NAC, SOD and catalase significantly rescued HeLa cells from pyrogallol-induced apoptosis accompanied by the recovery of GSH depletion. PMID- 18506366 TI - Regulation of Alstrom syndrome gene expression during adipogenesis and its relationship with fat cell insulin sensitivity. AB - Alstrom syndrome (ALMS) is an autosomal recessive genetic disease with characteristic phenotypical features including multi-organ fibrosis, insulin resistance, obesity and type 2 diabetes. ALMS1, a ubiquitously expressed gene mutated in ALMS patients, gives rise to a protein of unknown function localized to basal bodies of ciliated cells and centrosomes. Together with Bardet-Biedl syndrome, ALMS is a member of genetic ciliopathies, but the link between cilia/centrosome deficits and metabolic abnormalities remains to be determined. In this study for the first time we quantified Alms1 expression in a cellular model of adipogenesis during the differentiation of 3T3-L1 cells. An early decrease in Alms1 mRNA was observed during preadipocyte to adipocyte conversion. However, acute treatment of preadipocytes with the adipogenic factors did not result in significant change of Alms1 expression. In addition, to study the possible relationship between Alms1 and the degree of fat cell insulin sensitivity, as assessed with an insulin-dependent 2-[1-3H]-deoxyglucose uptake assay, we induced either a reduction or an increase in 3T3-L1 adipocytes insulin sensitivity by a chronic treatment with insulin or rosiglitazone respectively. In all these conditions Alms1 expression remained unchanged. In conclusion, our results show that Alms1 is expressed at higher level in preadipocytes suggesting a role of the gene in the early phase of adipogenesis. Moreover, changes in fat cell insulin sensitivity do not imply any effect on Alms1 expression. PMID- 18506367 TI - Different sources of reactive oxygen species contribute to low potassium-induced apoptosis in cerebellar granule cells. AB - An early increase in ROS production is characteristic of cerebellar granule cells undergoing apoptosis in the presence of 5 mM KCl. However, the sources of this increase have not been investigated in detail. In particular whether there is a single enzymatic source or the increase in ROS production is the consequence of the involvement of different enzymes has not been studied in depth. Different enzymatic pathways may indeed contribute to the up-regulation of intracellular ROS production either directly or via side-chain reactions and a number of candidate enzymes are known to be involved in the apoptotic process in various cell types. The aim of this study was to identify the cellular sources of the ROS generated by CGCs undergoing apoptosis by low K+. A panel of specific inhibitors against phospholipase, cytochromes P450, cyclooxygenase, lipoxygenase, xanthine oxidase, ribonucleotide reductase and NADPH oxidase were used. We provide evidence that no single source of ROS can be identified in apoptotic CGCs, but the ROS generated through the arachidonic acid (AA) pathways, mainly via lipoxygenase activities, seems to be the most prominent. PMID- 18506368 TI - Identification of IRF6 gene variants in three families with Van der Woude syndrome. AB - Van der Woude syndrome is the most common cause of syndromic orofacial clefting. It is characterised by the presence of lip pits, cleft lip and/or cleft palate. It is transmitted in an autosomal dominant manner, with high penetrance and variable expressivity. Several mutations in the interferon regulatory factor 6 (IRF6) gene have been found in VWS families, suggesting that this gene is the primary locus. We screened for mutations in this gene in three families in our population. There was a recurrent nonsense mutation within exon 9 of the gene for a Malay family consisting of five affected members with different presentations. We also found a co-segregating rare polymorphism which would result in a non synonymous change 23 bases downstream of the nonsense mutation. This polymorphism was present in <1% of the Malay subjects screened, but was not found among the Chinese and Indians in our population. For another family, a 396C-->T mutation (R45W in the DNA-binding domain) was found in the proband, although the possibility of a genetic defect elsewhere could not be excluded because his mother and twin sister (both unaffected) also had this variant. In the third case with complete absence of family history, a de novo deletion spanning the whole IRF6 gene was detected in the child with VWS. This case of haploinsufficiency caused disruption of orofacial development but not other organ systems as the child has no other medical or developmental abnormalities despite the deletion of at least five other genes. PMID- 18506369 TI - Associations of the apolipoprotein A-I gene polymorphism and serum lipid levels in the Guangxi Hei Yi Zhuang and Han populations. AB - Hei Yi Zhuang is an isolated subgroup of the Zhuang minority in China. Little is known about the effects of the genetic variants on serum lipid profiles in this population. The present study was undertaken to estimate the effects of the apolipoprotein (apo) A-I gene polymorphism adjacent to the initiate transcription site (-75 bp G/A) on the serum lipid levels in the Hei Yi Zhuang and Han populations. A total of 474 subjects of Hei Yi Zhuang and 564 subjects of Han Chinese were surveyed by a stratified randomized cluster sampling. Serum lipid levels were measured, and apoA-I gene polymorphism determined by polymerase chain reaction and restriction fragment length polymorphism. The frequencies of G and A alleles were 70.25 and 29.75% in Hei Yi Zhuang, and 65.96 and 34.04% in Han (P<0.05), respectively. The genotypic frequencies in Han were significantly different between males and females, subjects with normal TG (< or =1.70 mmol/l) and those with high TG (>1.70 mmol/l), or subjects with normal apoA-I (> or =1.20 g/l) and those with abnormal apoA-I (<1.20 g/l; P<0.05-0.01), respectively. The levels of LDL-C and apoA-I in Hei Yi Zhuang were higher in GG genotype than in AA or GA genotype (P<0.05 for each), but the levels of TG was lower in AA genotype than in GA genotype (P<0.05). There were also significant differences in serum TG levels among the three genotypes in Hei Yi Zhuang (P<0.05). The levels of HDL-C in Han were higher in GG genotype than in AA genotype (P<0.05), but the levels of TG in Han were lower in GG genotype than in GA genotype (P<0.05). The levels of apoA-I in Hei Yi Zhuang and the levels of HDL-C and apoB in Han were significantly correlated with genotype (P<0.05 for all). Hypertriglyceridemia was negatively associated with genotype in Hei Yi Zhuang (P<0.01). There were significant differences in the apoA-I -75 bp G/A between the Hei Yi Zhuang and Han populations. An association of the apoAI -75 bp G/A and serum TG, LDL-C and apoA-I levels in Hei Yi Zhuang and serum TG, HDL-C and apoB levels in Han was also observed in this study. PMID- 18506370 TI - Induction of apoptosis by S-allylmercapto-L-cysteine, a biotransformed garlic derivative, on a human gastric cancer cell line. AB - Epidemiological and experimental carcinogenesis studies provide evidence that certain components of garlic have anti-cancer activity. Although the biotransformed garlic derivative S-allylmercapto-L-cysteine (SAMC) has been reported to show an inhibitory effect on tumorigenesis, the mechanisms are poorly understood. The present study investigated the effect of SAMC on the growth of human gastric cancer SNU-1 cells. Upon treatment with SAMC, a concentration dependent inhibition of cell proliferation was observed and cells developed many of the hallmark features of apoptosis, including DNA fragmentation and an increase in the sub-diploid population. The anti-proliferative and apoptotic effect of SAMC was associated with the induction of Bax, p53, and caspase-9, rather than the induction of Bcl-2 and p21. Mitochondrial cytochrome c activation and an in vitro caspase-3 activity assay demonstrated that the activation of caspases accompanies the apoptotic effect of SAMC, which mediates cell death. These results suggest that the apoptotic effect of SAMC on gastric cancer SNU-1 cells may be connected with caspase-3 activation through the induction of Bax and p53, rather then Bcl-2 and p21. PMID- 18506371 TI - Hormonal regulation of regucalcin mRNA expression in osteoblastic MC3T3-E1 cells. AB - The effect of various hormones on regucalcin mRNA expression in osteoblastic MC3T3-E1 cells in vitro was investigated. Cells with subconfluency were cultured for 24 or 48 h in a medium containing either vehicle or various hormones without fetal bovine serum. Regucalcin mRNA expression was significantly increased after culture with parathyroid hormone (synthetic human PTH; 10(-7) M), insulin-like growth factor-I (IGF-I; 10(-8) M), or 17beta-estradiol (10(-10) or 10(-9) M) for 48 h. Culture with 1,25-dihydroxyvitamin D3 (10(-7) M) for 48 h caused a significant decrease in regucalcin mRNA expression. Regucalcin mRNA expression was significantly decreased after culture with tumor necrosis factor-alpha (1 or 10 ng/ml of medium) for 24 or 48 h. The effect of PTH or IGF-I in increasing regucalcin mRNA expression was not seen in the presence of staurosporine (10(-8) M), an inhibitor of protein kinase C, or PD98059 (10(-7) M), an inhibitor of mitosis-activated protein kinase (MAP kinase), respectively, suggesting that regucalcin mRNA expression is enhanced through intracellular signaling factors. This study demonstrated that regucalcin mRNA expression in osteoblastic MC3T3-E1 cells is regulated by various hormones. PMID- 18506372 TI - Extracellular matrix secreted by senescent fibroblasts induced by UVB promotes cell proliferation in HaCaT cells through PI3K/AKT and ERK signaling pathways. AB - Chronic exposure to solar ultraviolet radiation (UV) induces photoaging, and ultimately photocarcinogenesis. Senescent human skin fibroblasts (HSFs) in UVB stress-induced premature senescence (UVB-SIPS) share a similar extracellular matrix (ECM) phenotype with other types of senescent fibroblast. ECM from senescent fibroblasts induced by a variety of stresses has been shown to promote preneoplastic and neoplastic epithelial cell growth, a potential mechanism in carcinogenesis. We undertook this study to explore whether the extracellular matrices from UVB-induced senescent fibroblasts have any effect on the proliferation of HaCaT cells. The results showed that ECM secreted from HSFs in UVB-SIPS has 13.15 and 29.27% more stimulatory effect on proliferation than ECM secreted from presenescent HSFs and non-ECM, respectively. ECM from fibroblasts in UVB-SIPS activates FAK, ERK, and AKT in HaCaT cells. ERK and PI3K/AKT inhibitors inhibit ECM-induced ERK, AKT activation and cell proliferation. Cytochalasin D, a destructive agent of the cytoskeleton, inhibits ECM-induced FAK activation and cell proliferation in HaCaT cells. Collectively, we conclude that ECM secreted from HSFs in UVB-SIPS promotes cell proliferation via ERK and PI3K/AKT pathways and modulation of FAK and cytoskeletal proteins in HaCaT cells. Pharmacological manipulation of those signaling components may lead to the prevention and treatment of skin cancer induced by chronic solar exposure. PMID- 18506373 TI - Effect of estrogen on bone resorption and inflammation in the temporomandibular joint cellular elements. AB - Several epidemiological studies have reported that temporomandibular disorder is more prevalent in women, which suggests the involvement of sex hormones, such as estrogen, in the pathogenesis of this disease. PCR amplification and Western blotting were employed to target the expression of estrogen receptors (ERs) in human fibroblast-like synovial and ATDC5 cells. The effect of estrogen was investigated through the expression of RANKL, osteoprotegerin (OPG), M-CSF/CSF-1 and c-fms. We showed expression of M-CSF/ CSF-1 and c-fms, with time-dependent increase in both after the addition of estrogen. Based on previous studies reporting that M-CSF/CSF-1 regulates the proliferation and differentiation of hemopoietic progenitor cells into mature macrophages, we put forward a new hypothesis based on the increased inflammation and tendency of females to suffer more from temporomandibular disorder (TMD) in the presence of external exacerbating factors. Detection of RANKL and OPG in ATDC5 and expression of both in HFLS was confirmed with complete disappearance of the RANKL band, and marked increase in the expression of OPG after 1 h from the addition of estrogen. PMID- 18506374 TI - HBV and HCV infection in Japanese dental care workers. AB - Protective measures against occupational exposure to the hepatitis B virus (HBV) and hepatitis C virus (HCV) must be taken in order to prevent infection in dental care workers. To determine the best way to protect these workers, our study examined viral hepatitis infection in dental care workers in regions with a high prevalence of HCV infections in Japan. In total, 141 dental care workers (including dentists, dental hygienists and dental assistants) were enrolled. After a questionnaire to elicit demographic information was administered by an oral surgeon, hepatitis B surface antigen (HBsAg), antibody to HBs (anti-HBs), antibody to hepatitis B core antigen (anti-HBc) and antibody to HCV (anti-HCV) were measured. When necessary, HBeAg, anti-HBe, levels of HBV DNA, anti-HBc IgM and HCV RNA in serum were measured. Of the dental care workers included, 68 (48.2%) had been immunized with a HBV vaccine. Only 9 wore a new pair of gloves for each new patient being treated, 36 changed to a new pair only after the old gloves were torn and 24 did not wear any gloves at all. No one was positive for HBsAg or anti-HCV, though 73 (51.8%) and 17 (12.1%) workers were respectively positive for anti-HBs and anti-HBc. The positive rate of anti-HBc varied directly with worker age and experience. Of the 68 workers immunized with HBV vaccine, 51 (75%) were positive for anti-HBs. Of the 63 workers who were not so immunized, 17 (27%) were positive for anti-HBs and 15 of these were also positive for anti-HBc. Immunized workers were more protected against HBV infection than non-immunized workers, indicating that HBV vaccine was a useful measure for protection against the infection. The anti-HBc positive rate was significantly higher among dental care workers than general blood donors, suggesting that frequency of exposure to HBV was greater in dental care workers. HBV vaccination should be made compulsory for all dental care workers who handle sharp instruments. PMID- 18506375 TI - Association of genetic variants with atherothrombotic cerebral infarction in Japanese individuals with metabolic syndrome. AB - Metabolic syndrome is a risk factor for cardiovascular disease. The aim of the present study was to identify genetic variants that confer susceptibility to atherothrombotic cerebral infarction among individuals with metabolic syndrome in order to allow prediction of genetic risk for this condition. The study population comprised 1284 unrelated Japanese individuals with metabolic syndrome, including 313 subjects with atherothrombotic cerebral infarction and 971 controls. The genotypes for 296 polymorphisms of 202 candidate genes were determined with a method that combines the polymerase chain reaction and sequence specific oligonucleotide probes with suspension array technology. The Chi-square test, multivariable logistic regression analysis with adjustment for age, sex, body mass index, and the prevalence of hypertension, hypercholesterolemia, and diabetes mellitus, as well as a stepwise forward selection procedure revealed that the 2445G-->A (Ala54Thr) polymorphism (rs1799883) of FABP2, the -108/3G-->4G polymorphism of IPF1 (S82168), the A-->G (Thr94Ala) polymorphism (rs2241883) of FABP1, the G-->A (Asp2213Asn) polymorphism (rs529038) of ROS1, the -11377C-->G polymorphism (rs266729) of ADIPOQ, the 162A-->C polymorphism (rs4769055) of ALOX5AP, the -786T-->C polymorphism (rs2070744) of NOS3, and the 3279C-->T polymorphism (rs7291467) of LGALS2 were associated (P<0.05) with the prevalence of atherothrombotic cerebral infarction. Among these polymorphisms, the 2445G-->A (Ala54Thr) polymorphism of FABP2 was most significantly associated with this condition. Our results suggest that FABP2, IPF1, FABP1, ROS1, ADIPOQ, ALOX5AP, NOS3, and LGALS2 are susceptibility loci for atherothrombotic cerebral infarction among Japanese individuals with metabolic syndrome. Genotypes for these polymorphisms, especially for the 2445G-->A (Ala54Thr) polymorphism of FABP2, may prove informative for the prediction of genetic risk for atherothrombotic cerebral infarction among such individuals. PMID- 18506377 TI - PPARdelta increases expression of the human apolipoprotein A-II gene in human liver cells. AB - The peroxisome proliferator-activated receptor delta (PPARdelta) is a transcription factor that regulates genes of importance in lipid and glucose metabolism. ApoA-II is one of the major proteins of the HDL-particle. The aim of this study was to investigate the regulation of apoA-II gene expression by PPARdelta. Treatment of HepG2 cells with the PPARdelta specific agonist GW501516 increased apoA-II mRNA expression. Likewise, reporter gene assays using a construct containing 2.7 kb of the proximal apoA-II promoter showed increased activity after treatment with GW501516, both in HepG2 and in HuH-7 cells. Mutation of two putative PPAR response elements (PPREs) in this region showed that the PPRE at position -737/-717 is the functional site. Binding of PPARdelta to this site was confirmed by chromatin immunoprecipitation and gel retardation analyses. In conclusion, PPARdelta increases the expression of the human apoA-II gene in liver cells via a PPRE in the proximal promoter. PMID- 18506376 TI - Validation of gefitinib effectiveness in a broad panel of head and neck squamous carcinoma cells. AB - Recently improved understanding of the pathogenesis of human head and neck squamous cell carcinoma (HNSCC) has led to the development of new, molecular based therapeutic strategies, one of the more promising is the utilisation of tyrosine kinase (TK) inhibitors, targeting epidermal growth factor receptor (EGFR). In this study, we tested for gefitinib effectiveness in a broad panel of 12 newly established HNSCC cell lines, investigating its ability to reduce cell growth, to induce apoptosis and to modulate cell cycle and various EGFR pathway related targets. Gefitinib IC50 values ranged between 0.064 and 33 microM, its capability to induce apoptosis and cell accumulation in G0/G1 phase was cell line specific, and the main EGFR-related pathway involved in gefitinib activity was PI3K/Akt/mTor. We characterised our in vitro panel extensively, with the aim to identify predictive factors for gefitinib effectiveness; all cell lines were free of human papillomavirus infection, two were positive for Fhit expression, four expressed wild-type p53, and all of them variously expressed the other two p53 family members, p63 and p73. The comparison between the targets analysed and gefitinib effectiveness evidenced the absence of a clear relationship, excluding them as predictive factors for gefitinib efficacy. Our results confirmed the in vitro efficacy of an anti-EGFR approach, but other targets than those analysed here should be characterised in order to identify valid predictive factors for gefitinib utilisation. PMID- 18506378 TI - Murine intramyocellular lipids quantified by NMR act as metabolic biomarkers in burn trauma. AB - It has been suggested that intramyocellular lipids (IMCLs) may serve as biomarkers of insulin resistance and mitochondrial dysfunction. Using a hind-limb mouse model of burn trauma, we tested the hypothesis that severe localized burn trauma involving 5% of the total body surface area causes a local increase in IMCLs in the leg skeletal muscle. We quantified IMCLs from ex vivo intact tissue specimens using High-Resolution Magic Angle Spinning (HRMAS) 1H NMR and characterized the accompanying gene expression patterns in burned versus control skeletal muscle specimens. We also quantified plasma-free fatty acids (FFAs) in burn versus control mice. Our results from HRMAS 1H NMR measurements indicated that IMCL levels were significantly increased in mice exposed to burn trauma. Furthermore, plasma FFA levels were also significantly increased, and gene expression of Glut4, insulin receptor substrate 1 (IRS1), glycolytic genes, and PGC-1beta was downregulated in these mice. Backward stepwise multiple linear regression analysis demonstrated that IMCL levels correlated significantly with FFA levels, which were a significant predictor of IRS1 and PGC-1beta gene expression. We conclude from these findings that IMCLs can serve as metabolic biomarkers in burn trauma and that FFAs and IMCLs may signal altered metabolic gene expression. This signaling may result in the observed burn-induced insulin resistance and skeletal muscle mitochondrial dysfunction. We believe that IMCLs may therefore be useful biomarkers in predicting the therapeutic effectiveness of hypolipidemic agents for patients with severe burns. PMID- 18506379 TI - Multivariate and Geoaccumulation Index evaluation in mangrove surface sediment of Mengkabong lagoon, Sabah. AB - Spatial variations in estuarine intertidal sediment have been often related to such environmental variables as salinity, sediment types, heavy metals and base cations. However, there have been few attempts to investigate the difference condition between high and low tides relationships and to predict their likely responses in an estuarine environment. This paper investigates the linkages between environmental variables and tides of estuarine intertidal sediment in order to provide a basis for describing the effect of tides in the Mengkabong lagoon, Sabah. Multivariate statistical technique, principal components analysis (PCA) was employed to better interpret information about the sediment and its controlling factors in the intertidal zone. The calculation of Geoaccumulation Index (I(geo)) suggests the Mengkabong mangrove sediments are having background concentrations for Al, Cu, Fe, and Zn and unpolluted for Pb. Extra efforts should therefore pay attention to understand the mechanisms and quantification of different pathways of exchange within and between intertidal zones. PMID- 18506380 TI - An assessment of mercury loading in core sediments of Sunderban mangrove wetland, India (a preliminary report). AB - This is a preliminary report on total mercury (T(Hg)) in core sediments (<63 microm particle size) of Sunderban mangrove wetland, northeastern part of the Bay of Bengal, India. Cold vapor atomic absorption spectrometry (CVAAS) was used for T(Hg) determination. The concentration varies from 9.8 to 535.1 ppb (ngg(-1)). Results revealed variations over premonsoon and postmonsoon month at different core depth, as well as in studied three sampling stations, located at the site of three rivers: Hugli River (S(1)), Matla River (S(2)) and Bidyadhari River (S(3)). Elevated concentration of T(Hg) in subsurface layer (4-8 cm) of the core at S(2) is attributed to remobilization of mercury from deeper sediment (32-36 cm). Positive correlation is present between total Hg and clay content. Based on index of geoaccumulation (I(geo)) and Effects-Range Low (ER-L) value (150 ppb) it is considered that the sediments are till now unpolluted. As a consequence, there is less chance of ecotoxicological risk to organisms living in studied sediments. Two statistical methods were applied to determine T(Hg) anomalies. Box plot method showed one extreme and three outliers in S(1) at postmonsoon season. Two extremes were found at S(2) at 4-8 and at 32-36 cm in premonsoon period. In S(3) there was no anomaly by box plot method. MAD method was more sensitive than box plot method and T(Hg) anomaly was detected at 12-16 cm in S(3) during postmonsoon season. The data reported are useful baselines for T(Hg) in Sunderban mangrove wetland, India and would be of help in future sediment quality studies. PMID- 18506381 TI - Pesticide residues in honey samples from Himachal Pradesh (India). AB - Honey, being a natural product manufactured by honey bees is considered to be free from any extraneous material. The over-reliance on pesticides caused several environmental problems including pesticide residues in food. This constitutes a potential risk for human health, because of their sub acute and chronic toxicity. Therefore this study was carried out to know the extent of pesticide residue in honey produced in the various parts of Himachal Pradesh. Among different pesticides analysed in honey; HCH and its isomers were the most frequently detected followed by DDT and its isomers. Of the studied synthetic pyrethroids, only cypermethrin was found in honey samples. Residues of organophosphates viz. acephate, chlorpyriphos, ethion and monocrotophos were not detected, however malathion's residue was found exceeding the MRL (5 ppb) proposed by the Ministry of Commerce, Government of India. More over honey from natural vegetation contained lesser residues. It can be concluded that honey from Himachal Pradesh had low pesticide residues. PMID- 18506382 TI - Incontinence pads: recommending the best product-based wetback performance and price. AB - Incontinence pads are available in the USA without a prescription and are commonly the first treatment option a patient with incontinence uses. The goal of this study was to examine the difference in the performance and cost of commercially available incontinence pads with the intention of providing recommendations to women. Ten different urinary incontinence products were selected. A modified wetback test was used to test product performance. For the small volume leaks, the Walgreen's Extra pad generally performed worse on the wetback test than the three other pads tested (p = 0.001-0.012), but four tests were not statistically significant. At larger leak volumes, the Walgreen's underwear generally performed worse than other products (p < or = 0.001-0.046), with some exceptions. Brand name products generally performed better than generic products, but cost more. Undergarments and underwear do the worst job of keeping moisture inside the pad. PMID- 18506383 TI - The Portuguese validation of the International Consultation on Incontinence Questionnaire-Vaginal Symptoms (ICIQ-VS) for Brazilian women with pelvic organ prolapse. AB - The aim of this study is to validate the International Consultation on Incontinence Questionnaire-Vaginal Symptoms (ICIQ-VS) in Portuguese. Two hundred four women (108 symptomatic, 94 asymptomatic, and two with no data) with mean age of 55.4 years received a Portuguese version of the ICIQ-VS. Clinical data and pelvic organ prolapse quantification index (POP-Q) were obtained. Retest was performed 3 weeks later. Responsiveness was assessed after 20 weeks of postsurgical follow-up. Overall, most patients presented POP-Q > 2. ICIQ-VS demonstrated good psychometric properties (validity, reliability and responsiveness). The test-retest reliability was moderate to excellent for all questions. The construct validation distinguished differences in ICIQ-VS scores between symptomatic (ICIQ-VS5a > 0) and asymptomatic (ICIQ-VS5a = 0) women. ICIQ VS was highly responsive to surgical treatment and discriminated between levels of change in the vaginal symptoms score, sexual matters score, quality-of-life score, and POP-Q. The Portuguese version of ICIQ-VS was successfully validated. PMID- 18506384 TI - Water solution of onion crude powder inhibits RANKL-induced osteoclastogenesis through ERK, p38 and NF-kappaB pathways. AB - Onion powder has been reported to decrease the ovariectomy-induced bone resorption of rats. However, the molecular mechanism of onion powder on the bone cells has not been reported. Here, we report that water solution of onion crude powder decreases the osteoclastogenesis from co-cultures of bone marrow stromal cells and macrophage cells. Additionally, water solution of onion crude powder inhibits the RANKL-induced ERK, p38 and NF-kappaB activation in macrophages. In summary, our data showed that onion powder may benefit bone through an anti resorption effect on the osteoclasts. INTRODUCTION: A nutritional approach is important for both prevention and treatment of osteoporosis. Onion has been reported to decrease the ovariectomy-induced bone resorption. However, the functional effects of onion on the cultured osteoclasts and osteoblasts remain largely unknown. Here, we found that water solution of onion crude powder markedly inhibited the receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis through ERK, p38 and NF-kappaB pathways. Other studies were also designed to investigate the potential signaling pathways involved in onion-induced decrease in osteoclastogenesis. METHODS: The osteoclastogenesis was examined using the TRAP staining method. The MAPKs and NF kappaB pathways were measured using Western blot analysis. A transfection protocol was used to examine NF-kappaB activity. RESULTS: Water solution of onion crude powder inhibited the RANKL plus M-CSF-induced osteoclastic differentiation from either bone marrow stromal cells or from RAW264.7 macrophage cells. Treatment of RAW264.7 macrophages with RANKL could induce the activation of ERK, p38 and NF-kappaB that was inhibited by water solution of onion crude powder. On the other hand, it did not affect the cell proliferation and differentiation of human cultured osteoblasts. CONCLUSIONS: Our data suggest that water solution of onion crude powder inhibits osteoclastogenesis from co-cultures of bone marrow stromal cells and macrophage cells via attenuation of RANKL-induced ERK, p38 and NF-kappaB activation. PMID- 18506386 TI - Decreased plasma levels of soluble receptor for advanced glycation end products in mild cognitive impairment. AB - Growing evidence advanced the idea that the soluble form of the receptor for advanced glycation end-products (sRAGE) might serve as a risk marker for several disorders including Alzheimer disease. We found a reduced level of circulating sRAGE in patients with mild cognitive impairment (MCI). The reduction of sRAGE in MCI, as well as the anticipation of the disease in patients with the lowest sRAGE levels ( 0.05). Furthermore, digital tooth-shade measurements revealed no statistically-significant group-associated differences between those areas treated with bonding material and those not so treated, nor between bonded and debonded areas (p > 0.05). Nor did we observe any statistically-significant differences between bleached areas and those areas bleached after bonding and debonding procedures (p > 0.05). CONCLUSION: The processes of bonding and debonding alone do not seem to have any statistically-significant influence on the tooth color of bovine enamel using these testing materials, nor does the subsequent bleaching procedure. PMID- 18506403 TI - Changes in arch form following surgically-assisted rapid maxillary expansion (SRME). AB - BACKGROUND AND AIM: Surgically-assisted rapid maxillary expansion is a frequently employed procedure for expanding the maxilla in adults to treat pronounced transversal discrepancies of the upper jaw. Several studies have shown how the position of the anchorage teeth changes due to SRME. However, there is little detailed information available on the transversal movement of each tooth, its change in inclination and subsequent alterations of the arch length, width and sagittal arch form. MATERIAL AND METHODS: In our study we investigated those changes in 31 patient casts after SRME, following the active phase, and after a 3 month retention period whereby the Hyrax appliance remained blocked in situ. The casts were examined using a 3-dimensional reflex microscope. RESULTS: After active treatment the second molars moved 28% less than the amount of total expansion. At 9.6 degrees (first bicuspid) and 11.6 degrees (first molar), the anchorage teeth were also more buccally tipped than the second molars (7.4 degrees ). Unlike with conventional maxillary expansion, the measured increase in arch length here corresponded to the amount of anchorage teeth expansion. The sagittal arch decreased significantly during the retention phase in those teeth in front of and behind the anchorage teeth. Three months later we found that only 68% of the original expansion had occurred in the canines and second molars. The inclination of the teeth decreased slightly but significantly. We observed an only temporary increase in the sagittal dimension after appliance activation. There was an average loss of 0.83 mm at the end of the retention phase. Moreover, the anterior dental arch length only experienced a 50% increase in space over the expansion achieved initially. CONCLUSION: To minimize relapse rates in cases of severe crowding, we recommend that the canines and second molars be included in the appliance. PMID- 18506404 TI - Cephalometric analysis of molar and anterior tooth movement during cervical headgear treatment in relation to growth patterns. AB - OBJECTIVE: In this study we aimed to identify the reasons why cervical headgear treatment fails to induce bite opening or clockwise rotation of the mandible in the presence of a vertical growth pattern. In so doing, we investigated the dental effects of headgear treatment relative to growth patterns. MATERIALS AND METHODS: Records of 119 patients were analyzed, including casts and lateral cephalograms that had been obtained immediately before and after isolated cervical headgear treatment. Dental changes were determined relative to the SN line and a perpendicular line through point S. Inclusion criteria were: improvements in occlusal relationship of at least 4 mm in molar segments and treatment lasting at least 6 months. An orthodontic force of 3.5-4 N was applied in all cases through headgear featuring non-angulated outer arms. The headgear was worn for 14 hours a day during the active treatment phase. At the beginning of headgear treatment, patients were allocated into six groups according to y axis values to quantify growth-related differences. RESULTS: The upper first molar was moved an average of 1.3 mm distally, while the lower first molar was moved an average of 2 mm mesially. These movements revealed no differences in terms of growth patterns. We were not able to make representative findings pertaining to upper molar extrusion due to distal tilting and the measuring point's position. We used the lower molars' movements for analysis instead, which demonstrated mean downward changes of 5.1 mm in all groups. The vertical growth pattern groups presented smaller caudal movements in the upper and lower anterior segments than did those groups with horizontal growth pattern. The occlusal plane's inclination decreased more extensively in the vertical growth pattern groups than in those with a horizontal pattern (e.g. H2: -0.1 degrees ; V3: -2.3 degrees ). CONCLUSION: Our results suggest that while the upper molars extrude, the mandible rotates around the molars in a hypomochlion fashion in the presence of a vertical growth pattern, thus leading to bite deepening. PMID- 18506405 TI - A 57-year follow-up study of occlusion : part 1: oral health and attitudes to teeth among individuals with normal occlusion at the age of 8 years. AB - OBJECTIVE: To analyze occlusal changes between the ages of 8 and 65 years in 18 persons with normal occlusion at the age of 8 (N-group), to describe their attitudes toward dental esthetics and their experiences regarding their dentition. MATERIAL AND METHODS: The N-group is considered as the reference group in comparison with malocclusion groups that will be presented in subsequent articles. Documentation is based on intraoral photographs and personal interviews. RESULTS: In general, the occlusal changes were moderate. The probands were all satisfied with their teeth and tooth position. They were all examined at regular intervals by their dentists and practiced generally good oral hygiene. The average number of missing teeth was 1.4 (0-6) and the number of prosthodontically -restored or replaced teeth was 6.4 (0-24). Fifteen persons reported well-positioned teeth to be important. However, 17 found that visibly poor oral hygiene, discolored or missing teeth or the "total impression", to be more important than malpositioned front teeth. CONCLUSIONS: Generally speaking, occlusal changes were moderate and satisfaction with the dentition was good. Discolored or missing teeth and poor oral hygiene were found to be the most disturbing negative traits with respect to dental esthetics. PMID- 18506406 TI - The alignment of impacted and ectopic teeth using the Easy-Way-Coil (EWC) System. AB - BACKGROUND: Teeth that do not spontaneously erupt in the dental arch or are ectopic must normally be surgically exposed. For orthodontic correction a traction device must be attached to the affected tooth over which the already raised mucoperiosteal flap is lowered to improve the periodontal condition. METHOD: The author has developed a system at his private orthodontic practice for this purpose, and it has proven itself over a 10-year period. A traction spring is fastened to a lingual button that is attached to the etched tooth surface using an adhesive. The appliance is activated by systematically shortening the spring. RESULTS: A total of 103 patients were treated successfully in this way, 45 of whom had presented a unilaterally ectopic canine requiring an average treatment time of 17.8 months from surgical exposure to removal of the fixed appliance. None of the patients suffered inflammatory reactions in the oral mucosa adjacent to the traction spring during the treatment. Two impacted teeth were ankylosed and could not be aligned. CONCLUSION: The EWC-System is clinically a reliable, efficient, easy to use and economical system for the alignment of impacted and ectopic teeth. PMID- 18506407 TI - Letter to the editor : on Philipp Meyer-Marcotty, Jutta Hartmann, Angelika Stellzig-Eisenhauer: dentoalveolar open bite treatment with spur appliances. PMID- 18506408 TI - [Manufacture of autologous serum eye drops for out-patient therapy : cooperation between ophthalmic clinic and transfusion medicine department]. AB - BACKGROUND: Autologous serum eye drops are an important therapy option in severe ocular surface disorders and the therapeutic effectiveness has been demonstrated in many clinical studies. The production and use of autologous serum eye drops is strictly controlled by legal regulations in Germany: Both the German Medicines Act (AMG) and the Blood Transfusion Act regulate production, distribution and application, unless it is carried out by one person under controlled conditions in a hospital setting. MATERIAL AND METHODS: In cooperation with the ophthalmic clinic and the department of transfusion medicine, a standard operating procedure (SOP) was developed and a license for production and delivery of autologous serum eye drops was obtained from the appropriate local authorities. The experiences of the first two years of practice were analyzed. RESULTS: By an interfaculty cooperation, the possibility of legal and feasible out-patient treatment with autologous eye drops has been established at the University Hospital Erlangen. From 07/2005 to 07/2007, there ware 240 prescriptions for autologous serum eye drops. Unexpectedly, a relatively high rate (3.3%) of patients with primarily unknown viral or bacterial infectious diseases were found, which were diagnosed during the screening. These patients had to be excluded from autologous serum eye drop therapy. CONCLUSION: The treatment with autologous serum eye drops in an out patient setting is possible, when the infrastructure for manufacture and delivery is provided in accordance with existing regulations. PMID- 18506410 TI - [Medicine between humanitarianism and competition: the patients' point of view]. AB - BACKGROUND: An open medical market is supposed to be a promising tool for preserving the meagre resources of the German public health care system. The competition between humane and economic practice induces conflicts which burden physicians as well as patients. To analyse this problem by scientific means, inpatients were interviewed with the help of a standardised protocol. PATIENTS AND METHODS: During 10 weeks 524 inpatients were interviewed. The structured questionnaire consisted of nine closed-ended questions with multiple-choice answers. Question 3 included a free amendment. The gender ratio was balanced, and the age pattern represented the typical patient collective of our clinic. RESULTS AND CONCLUSION: The majority of patients expect negative changes in their medical service and the social attitude towards patients and physicians due to promoted medical competition. Besides an increasing financial load, losses in trust, self determination, and therapeutic freedom are expected. Instead a responsible transparency will not be achieved. Patients still judge the economically dominated reforms on the German health care market with scepticism or even hostility. In their point of view future discussions must aim at a more modern attitude towards health economics in order to bridge the schism they perceive between medical competition and humanitarianism. The understanding of strong financial management as a prime condition for stable social security is the basis of synergies to deal with upcoming reforms. PMID- 18506409 TI - Functional genomics and SNP analysis of human genes encoding proline metabolic enzymes. AB - Proline metabolism in mammals involves two other amino acids, glutamate and ornithine, and five enzymatic activities, Delta(1)-pyrroline-5-carboxylate (P5C) reductase (P5CR), proline oxidase, P5C dehydrogenase, P5C synthase and ornithine delta-aminotransferase (OAT). With the exception of OAT, which catalyzes a reversible reaction, the other four enzymes are unidirectional, suggesting that proline metabolism is purpose-driven, tightly regulated, and compartmentalized. In addition, this tri-amino-acid system also links with three other pivotal metabolic systems, namely the TCA cycle, urea cycle, and pentose phosphate pathway. Abnormalities in proline metabolism are relevant in several diseases: six monogenic inborn errors involving metabolism and/or transport of proline and its immediate metabolites have been described. Recent advances in the Human Genome Project, in silico database mining techniques, and research in dissecting the molecular basis of proline metabolism prompted us to utilize functional genomic approaches to analyze human genes which encode proline metabolic enzymes in the context of gene structure, regulation of gene expression, mRNA variants, protein isoforms, and single nucleotide polymorphisms. PMID- 18506411 TI - [Erionite-induced pleural plaques. Exposition to urban pollution in a female Turkish migrant in Germany]. AB - Erionite is a zeolite mineral of volcanic origin which contains no asbestos. It is found in different regions of the world, including southeast Turkey in ash and dust, from which it can cause inflammatory pleural plaques or malignant pleural mesothelioma (MPM). We report a female Turkish migrant exposed to urban pollution in her home country who decades later suffered from pleural plaques with a nonspecific chronic inflammatory disease. The differential diagnosis of inflammatory pleural plaques was assumed radiologically and confirmed by video assisted thoracoscopic biopsy. Short-term clinical and radiologic control of the patient will be necessary because of the risk of MPM. For epidemiologic considerations discussed referring to current literature, a growing incidence of this type of disease in migrants from high-risk areas must be reckoned with in Germany, even without exposition to asbestos. PMID- 18506412 TI - Interaction between dendritic cells and natural killer cells during pregnancy in mice. AB - A complex regulation of innate and adaptive immune responses at the maternal fetal interface promotes tolerance of trophoblast cells carrying paternally derived antigens. Such regulatory functions involve uterine dendritic cells (uDC) and natural killer (uNK) cells. The existence of a NK and DC "cross talk" has been revealed in various experimental settings; its biological significance ranging from cooperative stimulation to cell lysis. Little is known about the presence or role of NK and DC cross talk at the maternal fetal interface. The present study shows that mouse NK and DC interactions are subject to modulation by trophoblast cells in vitro. This interaction promotes a tolerogenic microenvironment characterized by downregulation of the expression of activation markers on uNK cells and uDC and dominance of Th2 cytokines. NK and DC interactions would also influence uterine cell proliferation and this process would be strongly modulated by trophoblast-derived signals. Indeed; while low proliferation rates were observed upon regular coculture allowing direct contact between uterine cells and trophoblasts, incubation in a transwell culture system markedly increased uterine cell proliferation suggesting that soluble factors are key mediators in the molecular "dialog" between the mother and the conceptus during the establishment of mouse pregnancy. Our data further reveal that the regulatory functions of trophoblast cells associated with tolerance induction are impaired in high abortion murine matings. Interestingly, we observed that secretion of interleukin-12p70 by uDC is dramatically abrogated in the presence of uNK cells. Taken together, our results provide the first evidence that a delicate balance of interactions involving NK cells, DC, and trophoblasts at the mouse maternal fetal interface supports a successful pregnancy outcome. PMID- 18506414 TI - [Cerclage using hollow screws for internal arthrodesis in cases of pronounced osteoporosis and osteopenia]. AB - Osteosynthesis of osteoporotic or osteopenic bone poses a considerable challenge for the surgeon, the type of osteosynthesis performed, and the osteosynthesis material employed. In osteoporotic and osteopenic bone, especially when circulation is decreased or sensitivity reduced, the principle of tension banding places great demands on the bone and surgeon since insufficient tightening of the wire results in inadequate immobilization while overtightening of the tension band can cause the wire to cut through the soft bone. This contribution describes a surgical technique, which prevents the wire from cutting through the bone. Cannulated screws are inserted vertically into the bone in the traction direction and the wire of the tension banding is passed through the cannulated screws. This ensures that the wire traction is distributed across the entire cross section of the bone. Thus, it is possible to completely tighten the tension band in osteopenic and osteoporotic bone. This technique is illustrated using the example of internal screw and tension band arthrodesis of the ankle joint, which achieved stable arthrodesis with weight bearing and recovered ambulation in a patient with pronounced diabetic foot syndrome and peripheral arterial occlusive disease. PMID- 18506413 TI - [First traumatic anterior shoulder dislocation: postoperative results 6 months and 2 years after arthroscopic stabilization in young patients]. AB - BACKGROUND: The purpose of this prospective clinical trial was to report about results of primary or early secondary arthroscopic stabilization after first traumatic anterior dislocation of the shoulder. PATIENTS AND METHODS: Within 2 weeks or in the 7th to 12th week post trauma, 51 subjects between 16 and 30 years received arthroscopic stabilization and rehabilitation. The patients were followed-up 6 months and 2 years post operation and assessed with the Rowe and the Constant scores. RESULTS: At a mean follow-up of 27.9+/-4.3 months all patients returned to work in their profession. Of the patients, 79.2% were satisfied with the result of the operation; 91.7% returned to their sports. During the follow-up period five patients (10.4%) suffered redislocation. There was no difference in the result comparing the time of operation. The Rowe and Constant scores showed excellent results. CONCLUSIONS: Arthroscopic stabilization after first traumatic anterior shoulder dislocation of the young patient is an appropriate approach and regardless of whether it is performed as a primary or early secondary operation it significantly lowers the redislocation rate. The method leads to quick reintegration into professional life and sports activities. PMID- 18506415 TI - Melanin- and carotenoid-dependent signals of great tits (Parus major) relate differently to metal pollution. AB - Due to their high phenotypic plasticity, the expression of secondary sexual characteristics is particularly sensitive to stress. Here, we investigated the expression of two conspicuous visual signals in great tits (Parus major) in a metal pollution gradient. In three study sites with marked differences in metal contamination (mainly lead, cadmium, copper and zinc), we compared melanin and carotenoid colouration of great tits. While carotenoid colouration (yellow breast) was negatively related to metal pollution, the size of a melanin trait (breast stripe) was larger in the most polluted sites. Environmental pollutants not only affect the expression of conspicuous signals but may even enhance, directly or indirectly, a signal of male quality such as breast stripe. Our results also support the multiple messages hypothesis predicting that different signals highlight different aspects of geno- and phenotypic condition of the bearer. PMID- 18506417 TI - Molecular mapping of soybean rust (Phakopsora pachyrhizi) resistance genes: discovery of a novel locus and alleles. AB - Soybean production in South and North America has recently been threatened by the widespread dissemination of soybean rust (SBR) caused by the fungus Phakopsora pachyrhizi. Currently, chemical spray containing fungicides is the only effective method to control the disease. This strategy increases production costs and exposes the environment to higher levels of fungicides. As a first step towards the development of SBR resistant cultivars, we studied the genetic basis of SBR resistance in five F2 populations derived from crossing the Brazilian-adapted susceptible cultivar CD 208 to each of five different plant introductions (PI 200487, PI 200526, PI 230970, PI 459025, PI 471904) carrying SBR-resistant genes (Rpp). Molecular mapping of SBR-resistance genes was performed in three of these PIs (PI 459025, PI 200526, PI 471904), and also in two other PIs (PI 200456 and 224270). The strategy mapped two genes present in PI 230970 and PI 459025, the original sources of Rpp2 and Rpp4, to linkage groups (LG) J and G, respectively. A new SBR resistance locus, rpp5 was mapped in the LG-N. Together, the genetic and molecular analysis suggested multiple alleles or closely linked genes that govern SBR resistance in soybean. PMID- 18506416 TI - [Treatment of arteriosclerotic diseases of the carotid artery. The carotid stent]. AB - In order to find the most suitable therapy concept for patients with severe stenosis of the carotid bifurcation (>70%), knowledge on the currently available treatment methods is necessary. In addition to carotid endarterectomy, medication therapy and stent angioplasty are also available. The outcome after stent angioplasty must be considered under the aspects of rapid technical development and increasing experience. Results of studies are still only of limited value with respect to the development of protection systems, flexible guiding catheters, low profile and more visible stents and balloons as well as using thrombocyte aggregation. PMID- 18506418 TI - QTL analysis of cleistogamy in soybean. AB - Early-maturing cultivars of soybean [Glycine max (L.) Merr.] native to the shores of the Sea of Okhotsk (Sakhalin and Kuril Islands) and eastern Hokkaido (northern Japan) have a strong tendency to produce cleistogamous flowers throughout their blooming period. A previous study revealed that cleistogamy is controlled by a minimum of two genes with epistatic interaction, one of which is associated with a maturity gene responsible for insensitivity to incandescent long daylength (ILD). This study was conducted to determine the genetic basis of cleistogamy in more detail by QTL mapping. F2 to F4 progenies derived from a cross between a cleistogamous cv. Karafuto-1 and a chasmogamous cv. Toyosuzu were used. A molecular linkage map spanning 2,180 cM comprising 500 markers was constructed using 89 F2 plants. The markers were distributed in 25 linkage groups. An interval mapping method to analyze categorical traits identified four QTLs for cleistogamy, cl1, cl2, cl3 and cl4, in molecular linkage groups (MLGs) C2, D1a, I and L, respectively. Alleles derived from Karafuto-1 had additive effects to increase probability of cleistogamy at cl3 and cl4, whereas the alleles had additive effects to decrease the probablity at cl1 and cl2. Progeny test confirmed the effects of cl3, which had the highest LOD score (5.20). Composite interval mapping revealed four QTLs for flowering date, fd5-fd8. Judging from relative location with markers and association with ILD responses, fd7 and fd8 may correspond to maturity genes E4 and E3, respectively. cl3 and cl4 were located at similar positions as fd7 and fd8, suggesting that the two maturity genes may control cleistogamy by either pleiotropy or close linkage. PMID- 18506419 TI - Bryan Jennett and the field of traumatic brain injury. His intellectual and ethical heritage in neuro-intensive care. AB - William Bryan Jennett, one of the leading figures in neurosurgery of the twentieth century, has died on 26 January 2008, at the age of 81. He made fundamental contributions to the field of traumatic brain injury (TBI) that still shape diagnosis, management and prognosis worldwide, in the second part of the last century. This paper is meant to gratefully acknowledge his contributions and to reflect on the implications that his work has for neurointensive care today. Starting from his early steps, we tried to highlight his fundamental work on diagnosis of severity in TBI, on rescue, treatment and prognosis of severe TBI. Moreover, his contribution in the definition of vegetative state, minimally conscious state and brain death has been emphasized. The contribution of Professor Bryan Jennett was in fact seminal in many aspects: the application of a common language in brain damage evaluation, where GCS and GOS are now universally employed; a critical approach to TBI diagnosis and treatment, in the search of proven better therapies; a quantitative approach to TBI prognosis, based on large clinical series and appropriate statistics; a strong commitment to the ethical implication of survival after severe injury, including the vegetative status; social responsibility in the diagnosis of brain death and in organ donors procurement. For these reasons, he can be considered one of the leading figures in neurosurgery and neurology of the twentieth century. This paper is meant to gratefully acknowledge his contributions and to reflect on the implications that his work has for neuro-intensive care today. PMID- 18506421 TI - Selective decontamination is not sticky. PMID- 18506422 TI - Sulfoacetate released during the assimilation of taurine-nitrogen by Neptuniibacter caesariensis: purification of sulfoacetaldehyde dehydrogenase. AB - Taurine (2-aminoethanesulfonate) is a widespread natural product whose nitrogen moiety was recently shown to be assimilated by bacteria, usually with excretion of an organosulfonate via undefined novel pathways; other data involve transcriptional regulator TauR in taurine metabolism. A screen of genome sequences for TauR with the BLAST algorithm allowed the hypothesis that the marine gammaproteobacterium Neptuniibacter caesariensis MED92 would inducibly assimilate taurine-nitrogen and excrete sulfoacetate. The pathway involved an ABC transporter (TauABC), taurine:pyruvate aminotransferase (Tpa), a novel sulfoacetaldehyde dehydrogenase (SafD) and exporter(s) of sulfoacetate (SafE) (DUF81). Ten candidate genes in two clusters involved three sets of paralogues (for TauR, Tpa and SafE). Inducible Tpa and SafD were detected in cell extracts. SafD was purified 600-fold to homogeneity in two steps. The monomer had a molecular mass of 50 kDa (SDS-PAGE); data from gel filtration chromatography indicated a tetrameric native protein. SafD was specific for sulfoacetaldehyde with a K (m)-value of 0.12 mM. The N-terminal amino acid sequence of SafD confirmed the identity of the safD gene. The eight pathway genes were transcribed inducibly, which indicated expression of the whole hypothetical pathway. We presume that this pathway is one source of sulfoacetate in nature, where this compound is dissimilated by many bacteria. PMID- 18506423 TI - Variations in the multiple tbp genes in different Halobacterium salinarum strains and their expression during growth. AB - The presence and expression of the multiple tbp genes encoding TATA-box binding proteins (TBPs) was investigated in various strains and mutants of the archaeon Halobacterium salinarum. Six genes, tbpA through tbpF, are present in the genome of Hbt. salinarum NRC-1 and also in the gas vesicle negative mutant strain R1. The only tbp gene located in the chromosome is tbpE, whereas all others are found in the plasmid DNA. Due to the dynamic nature of the plasmids in the Halobacterium strains, the copy numbers of the alternative tbp genes vary significantly. Five tbp genes (tbpA through tbpE) were present in the wild-type strain Hbt. salinarum PHH1. The tbpC gene of Hbt. salinarum PHH1 carried an ISH27 2 insertion element at the start of the reading frame that prevented the expression. All other tbp genes of PHH1 were expressed under aerobic and anaerobic growth conditions and quantitative RT-PCR yielded tbpE as dominant tbp transcript during the exponential growth phase. The plasmid deletion variant Hbt. salinarum PHH4 lacked all of the tbp genes except for tbpE and showed an altered growth behaviour compared to PHH1 wild-type in the stationary growth phase under anaerobic growth conditions. PMID- 18506424 TI - Astrocytic alkalinization by therapeutically relevant lithium concentrations: implications for myo-inositol depletion. AB - RATIONALE: One theory for therapeutic effects of the lithium ion (Li+) in bipolar disorder is that myo-inositol, needed for phospholipase C-mediated signaling, is depleted by Li(+)-induced inhibition of inositolphosphate hydrolysis or of myo inositol uptake, an effect demonstrated in cultured mouse astrocytes at high myo inositol concentrations. In contrast, myo-inositol uptake is inhibited at low concentrations, reflecting that it occurs both by the high-affinity Na(+) dependent myo-inositol transporter (SMIT) and the lower-affinity H(+)-dependent inositol transporter (HMIT). Increased intracellular pH (pHi) stimulates SMIT but inhibits HMIT, suggesting that the effect of Li+ could be caused by intracellular alkalinization. In this study, we therefore investigated Li+ effects on intracellular pH in astrocytes, measured by 2',7'-bis(2-carboxyethyl)-5(6) carboxyfluorescein (BCECF) fluorescence. RESULTS: Chronic treatment with the therapeutically relevant Li+ concentration of 1 mM for 2 or 3 weeks increased pHi by approximately 0.10, whereas 0.5 mM was ineffective, and 2 mM caused a larger increase. The alkalinization resulted from acute stimulation of the Na+/H+ exchanger (NHE) by extracellular Li+, demonstrated after acid load with NH4Cl. In response to continuous stimulation, NHE1 mRNA was down-regulated, but protein was not. CONCLUSIONS: Chronic treatment with pharmacologically relevant Li+ concentrations increases pHi in astrocytes, creating conditions for decreased uptake of high myo-inositol concentrations and increased uptake of low concentrations. The pharmacological relevance of this effect is supported by literature data suggesting brain acidosis in bipolar patients and by preliminary observations that carbamazepine and valproate also increase pHi in astrocytes. Stimulation of NHE1-stimulated sodium ion uptake might also trigger uptake of chloride ions and osmotically obliged water. PMID- 18506426 TI - Synthesis, characterization, and high-performance liquid chromatographic evaluation of C14 stationary phases containing branched and unbranched alkyl groups. AB - Reversed-phase materials with branched and unbranched alkyl groups were prepared by modifying porous, spherical silica gel in a two-step reaction-immobilization of a trifunctional alkoxysilane (3-glycidoxypropyltrimethoxysilane) on the silica surface followed by reaction with a branched and an unbranched octanoic acid. The chromatographic sorbents were characterized by solid-state (29)Si and (13)C NMR spectroscopy. The chromatographic behaviour of the stationary phases was evaluated by use of a test mixture according to the Standard Reference Material 870 set from the US National Institute of Standards and Technology, in order to study the effect of branched and unbranched alkyl chains. PMID- 18506425 TI - Acute tryptophan depletion in healthy males attenuates phasic cardiac slowing but does not affect electro-cortical response to negative feedback. AB - RATIONALE: Recent studies have shown that serotonin might be involved in performance monitoring, although the results have been inconclusive. Inconsistent results might be related to the type of pharmacological manipulation and the used behavioral and physiological measures. OBJECTIVES: The present study aimed at further specifying the role of serotonin in performance monitoring. MATERIALS AND METHODS: The effect of serotonin on performance monitoring was studied by using acute tryptophan depletion (ATD), a well-known method to transiently lower central serotonin levels. Twenty healthy male volunteers performed a time estimation task and their event-related brain potential (ERP), behavioral, and cardiac responses to feedback stimuli were measured. Furthermore, subjective mood and amino-acid levels were determined. RESULTS: As expected, ATD did not affect mood and lowered tryptophan levels. ATD attenuated cardiac slowing to negative feedback but did not affect responses to positive feedback, ERPs, and performance measures. CONCLUSIONS: The data point in the direction of a dissociation between cardiac and electro-cortical responses. Cardiac responses appear to be more sensitive to changes in serotonin metabolism and appear to reflect different aspects of the feedback stimulus. The phasic cardiac response appears to be an important measure that provides additional information about the impact of feedback stimuli and serotonergic functioning. PMID- 18506427 TI - A liquid chromatography method using a monolithic column for the determination of corticoids in animal feed and animal feeding water. AB - An HPLC-DAD method for determining corticoids in calf feed and in animal feeding water samples using a monolithic column has been developed and validated. The method optimization included the study of binary mobile phases of water and acetonitrile. The optimum separation was achieved at 40 degrees C, with acetonitrile:H(2)O 29:71 v/v used as mobile phase and a 3 ml/min flow-rate, which resulted in their separation in about 5 min. Two reported sample procedures were applied to feed and for animal feeding water samples prior to HPLC. Method validation was carried out according to the EU criteria established for quantitative screening methods. The results indicate that this method is highly specific, reproducible and accurate. The proposed method was found to be robust and unaffected by small variations in the extraction procedure and in HPLC conditions. The developed method for the determination of corticoids in feed and water samples was also found to be suitable for different kinds of feeds and waters. PMID- 18506428 TI - Analysis of amino acids without derivatization in barley extracts by LC-MS-MS. AB - A method has been developed for quantification of 20 amino acids as well as 13 (15)N-labeled amino acids in barley plants. The amino acids were extracted from plant tissues using aqueous HCl-ethanol and directly analyzed without further purification. Analysis of the underivatized amino acids was performed by liquid chromatography (LC)-electrospray ionization (ESI) tandem mass spectrometry (MS MS) in the positive ESI mode. Separation was achieved on a strong cation exchange column (Luna 5micro SCX 100A) with 30 mM ammonium acetate in water (solvent A) and 5% acetic acid in water (solvent B). Quantification was accomplished using d (2)-Phe as an internal standard. Calibration curves were linear over the range 0.5-50 microM, and limits of detection were estimated to be 0.1-3.0 microM. The mass-spectrometric technique was employed to study the regulation of amino acid levels in barley plants grown at 15 degrees C uniform root temperature (RT) and 20-10 degrees C vertical RT gradient (RTG). The LC-MS-MS results demonstrated enhanced concentration of free amino acids in shoots at 20-10 degrees C RTG, while total free amino acid concentration in roots was similarly low for both RT treatments. (15)NO(3) (-) labeling experiments showed lower (15)N/(14)N ratios for Glu, Ser, Ala and Val in plants grown at 20-10 degrees C RTG compared with those grown at 15 degrees C RT. PMID- 18506429 TI - Reverse transcription-polymerase chain reaction on a microarray: the integrating concept of "active arrays". AB - In this report we describe the proof of principle of a reverse transcription polymerase chain reaction (RT-PCR) but on-chip, with immobilized specific primers using a transcriptome of mouse-muscle fibroblasts for detection of muscle specific expression products of these cells. The isolated total mRNA was directly incubated on an array of immobilized and solubilized specific primers, which allow the amplification of certain muscle-specific RNAs via its immobilized cDNAs. In contrast to others, the immobilized cDNA-products were directly synthesized on the chip by applying covalently bound specific primers. The products were detected by the incorporated and fluorophore-modified specific primers of the subsequently synthezised second strand. In addition, this second strand served as a further template (like the basically used mRNA) in the subsequent solid-phase-PCR to amplify first-strand cDNA copies at the remaining immobilized specific primer-probes. This is the intrinsic factor of the amplification of certain signals of this application. The specific cDNA templates of genes coding for subunits of the mouse muscle acetylcholine receptor (Chrna1, Chrnb1, Chrnd) and the genes coding for myogenin (Myog), muscle creatine kinase (Ckmm), and ATPase (Atp2a2) were amplified on a biochip by RT-PCR directly from freshly isolated mRNA. The resulting procedure allows the detection of mRNA sequences from less than 5 pg of total RNA preparations. PMID- 18506430 TI - Development of a highly precise ID-ICP-SFMS method for analysis of low concentrations of lead in rice flour reference materials. AB - Microwave digestion and isotope dilution inductively coupled plasma mass spectrometry (ID-ICP-SFMS) has been applied to the determination of Pb in rice flour. In order to achieve highly precise determination of low concentrations of Pb, the digestion blank for Pb was reduced to 0.21 ng g(-1) after optimization of the digestion conditions, in which 20 mL analysis solution was obtained after digestion of 0.5 g rice flour. The observed value of Pb in a non-fat milk powder certified reference material (CRM), NIST SRM 1549, was 16.8 +/- 0.8 ng g(-1) (mean +/- expanded uncertainty, k = 2; n = 5), which agreed with the certified value of 19 +/- 3 ng g(-1) and indicated the effectiveness of the method. Analytical results for Pb in three brown rice flour CRMs, NIST SRM 1568a, NIES CRM 10-a, and NIES CRM 10-b, were 7.32 +/- 0.24 ng g(-1) (n = 5), 1010 +/- 10 ng g(-1) (n = 5), and 1250 +/- 20 ng g(-1) (n = 5), respectively. The concentration of Pb in a candidate white rice flour reference material (RM) sample prepared by the National Metrology Institute of Japan (NMIJ) was observed to be 4.36 +/- 0.28 ng g(-1) (n = 10 bottles). PMID- 18506432 TI - Spatial and temporal acuity of visual perception can be enhanced selectively by attentional set. AB - The aim of this research was to study the relationship between perceptual judgments about space and time. If spatial and temporal judgments were dissociable, they should be modulated selectively by attention. We compared the effect of the attentional set upon fine-grained spatial versus temporal discrimination of visual perception in two experiments. Using identical sensory stimulation, we measured perceptual judgments on either the size of a small spatial gap or the duration of a brief temporal gap. The attentional set was manipulated by cuing the task that was most likely to be performed. In one experiment, a neutral cue was also used, to measure relative benefits and costs of spatial and temporal task sets. If the attentional set could be directed selectively to spatial and temporal task-relevant dimensions, performance on both spatial and temporal acuity tasks should be specifically modulated by task cuing. The results showed that the attentional set enhanced the speed and accuracy of perceptual judgments similarly on both spatial and temporal tasks. Moreover, accuracy in one task was selectively enhanced by attending to that task while remaining unaffected by attending to the alternative task. This finding suggests multiple mechanisms, by which visual processing of spatial and temporal features can be selectively prepared without interfering with one another. PMID- 18506431 TI - Common synaptic input across motor nuclei supplying intrinsic muscles involved in the precision grip. AB - To gain insight into the mechanism by which the nervous system orchestrates the activities of multiple muscles during voluntary motor behaviors, we measured the degree of synchrony in the discharge times across pairs of motor units recorded from two intrinsic hand muscles, the adductor pollicis (AdP) and first dorsal interosseous (FDI), in human subjects performing a precision grip. The magnitude of synchrony measured across muscles is thought to reflect the extent of divergent synaptic input delivered in common to the motor nuclei supplying the two muscles. Unlike the pronounced motor-unit synchrony observed across comparable extrinsic hand muscles during the precision grip, little synchrony was detected across these two intrinsic muscles. These results suggest that extrinsic and intrinsic muscles of the hand are controlled by descending pathways with distinctly different patterns of spinal connectivity--more widespread terminations across motor nuclei supplying extrinsic muscles whereas more focal input to individual motor nuclei innervating intrinsic muscles. PMID- 18506433 TI - Reduced postural sway during quiet standing by light touch is due to finger tactile feedback but not mechanical support. AB - It is well known that a light and voluntary touch with a fingertip on a fixed surface improves postural stability during quiet standing. To determine whether the effect of the light touch is due to the tactile sensory input, as opposed to mechanical support, we investigated the light touch effect on postural stability during quiet standing with and without somatosensory input from the fingertip. Seven young subjects maintained quiet standing on a force platform with (LT) and without (NT) lightly touching a fixed surface, and with (TIS) and without (CON) the application of tourniquet ischemia, which removed the tactile sensation from the fingertip. The mean velocity of centre of pressure (CoP) was calculated to assess the postural sway in each condition. The mean velocity of CoP was significantly smaller in the LT condition compared to the NT condition only under the CON condition, whereas the light touch effect was not significant under the TIS condition. We found that the reduction of the horizontal ground reaction force due to the light touch was about 20%, which was approximately equivalent to the reduction of mean velocity of CoP in the LT condition compared to the NT condition. Since the fingertip contact force was relatively large compared to the horizontal ground reaction force, one could say that the light touch effect might be due to the mechanical support provided by the contact itself. However, we demonstrated experimentally that light touch effects were diminished due to loss of finger tactile feedback induced by the tourniquet ischemia, but not due to the mechanical support provided by the light touch. One possible reason is the lack of feedback information in controlling posture, and the other is the altered control of the arm induced by the loss of tactile feedback. PMID- 18506434 TI - Stimulus-driven selection of routes to imitation. AB - Several models have proposed that an action can be imitated via one of two routes: a direct visuospatial route, which can in principle mediate imitation of both meaningful (MF) and meaningless (ML) actions, and an indirect semantic route, which can be used only for MF actions. The present study investigated whether selection between the direct and indirect routes is strategic or stimulus driven. Tessari and Rumiati (J Exp Psychol Hum Percept Perform 30:1107-1116, 2004) have previously shown, using accuracy measures, that imitation of MF actions is superior to imitation of ML actions when the two action types are presented in separate blocks, and that the advantage of MF over ML items is smaller or absent when they are presented in mixed blocks. We first replicated this finding using an automated reaction time (RT), as well as accuracy, measure. We then examined imitation of MF and ML actions in the mixed condition as a function of the action type presented in the previous trial and in relation to the number of previous test trials. These analyses showed that (1) for both action types, performance was worse immediately after ML than MF trials, and (2) even at the beginning of the mixed condition, responding to MF actions was no better than responding to ML items. These results suggest that the properties of the action stimulus play a substantial role in determining whether imitation is mediated by the direct or the indirect route, and that effects of block composition on imitation need not be generated through strategic switching between routes. PMID- 18506435 TI - Natural textures classification in area V4 of the macaque monkey. AB - Natural texture of an object is an important cue for recognition. In real conditions, the incidence angle of light on natural textures leads to a complex pattern of micro-shading that modifies 3D rendering of surfaces. Little is known about visual processing of material properties. The present work aims to study the coding of natural textures by the neurons of area V4 of the awake macaque monkey. We used patches of natural textures issued from the CURET database and illuminated with two or three different angles with their corresponding controls (scrambled Fourier phase). We recorded the responses of V4 neurons to stimuli flashed in their receptive fields (RFs) while the macaques performed a simple fixation task. We show that a large majority of V4 neurons responded to texture patches with a strong modulation across stimuli. The analysis of those responses indicate that V4 neurons integrate first and second order parameters in the image (mean luminance, SNR, and energy), which may be used to achieve texture clustering in a multidimensional space. This clustering was comparable to that of a pyramid of Gabor filters and was not affected by illumination angles. Altogether, these results suggest that the V4 neuronal population acts as a set of filters able to classify textures independently of illumination angle. We conclude that area V4 contains mechanisms that are sensitive to the aspect of textured surfaces, even in an environment where illumination changes continuously. PMID- 18506436 TI - Plasma esterases and inflammation in ageing and frailty. AB - OBJECTIVES: Esterases are enzymes of drug metabolism known to be reduced in frail older people and during acute illness. The mechanism for this is unknown. The aim of this study was to examine esterase activity and inflammation in ageing and frailty. METHODS: Thirty frail patients (mean age 84.9 years) dependent on continuing inpatient care, 40 patients of intermediate frailty attending Day Hospital (84.2 years), 40 fit older controls (82.7 years) and 30 young controls (23.3 years) were studied. Frailty indicators, plasma esterase activities and markers of inflammation were measured. RESULTS: With increasing patient frailty, C-reactive protein (CRP), interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) increased significantly and esterase activity, with the exception of aspirin esterase, fell significantly (p < 0.005). There were significant negative correlations between log-transformed IL-6 and acetylcholinesterase (r = -0.354, p < 0.01), butyrylcholinesterase (r = -0.392, p < 0.01) and benzoylcholinesterase activity (r = -0.241, p < 0.05) and significant negative correlations between TNF alpha and acetylcholinesterase (r = -0.223, p < 0.01), butyrylcholinesterase (r = -0.279, p < 0.01) and benzoylcholinesterase activity (r = -0.253, p < 0.01). Aspirin esterase activity did not correlate with IL-6 or TNF- alpha. CONCLUSION: Frailty was associated with higher inflammatory markers and lower esterase activity. There was a weak but significant negative correlation between both IL-6 and TNF-alpha and the activity of three of four esterases. The negative correlation between esterase activity and inflammatory markers may have a causal basis, comparable to the inflammatory suppression of cytochrome P-450 enzymes. PMID- 18506438 TI - Closing the gap between coil and balloon in the neurointerventional armamentarium? Initial clinical experience with a nitinol vascular occlusion plug. AB - INTRODUCTION: The use of vascular plug devices for the occlusion of high-flow lesions is a relatively new and successful procedure in peripheral and cardiopulmonary interventions. We report on the use and efficiency of the Amplatzer vascular plug in a small clinical series and discuss its potential for occlusion of large vessels and high-flow lesions in neurointerventions. METHODS: Between 2005 and 2007 four patients (mean age 38.5 years, range 16-62 years) were treated with the device, in three patients to achieve parent artery occlusion of the internal carotid artery, in one patient to occlude a high-flow arteriovenous fistula of the neck. The application, time to occlusion, and angiographic and clinical results and the follow-up were evaluated. RESULTS: Navigation, positioning and detachment of the device were satisfactory in all cases. No flow related migration of the plug was seen. The cessation of flow was delayed by a mean of 10.5 min after deployment of the first device. In the procedures involving vessel sacrifice, two devices had to be deployed to achieve total occlusion. No patient experienced new neurological deficits; the 3-month follow up revealed stable results. CONCLUSION: The Amplatzer vascular plug can be adapted for the treatment of high-flow lesions and parent artery occlusions in the head and neck. In this small series the use of the devices was uncomplicated and safe. The rigid and large delivery device and the delayed cessation of flow currently limit the device's use in neurointerventions. PMID- 18506439 TI - Minimizing losses in bio-electrochemical systems: the road to applications. AB - Bio-electrochemical systems (BESs) enable microbial catalysis of electrochemical reactions. Plain electrical power production combined with wastewater treatment by microbial fuel cells (MFCs) has been the primary application purpose for BESs. However, large-scale power production and a high chemical oxygen demand conversion rates must be achieved at a benchmark cost to make MFCs economical competitive in this context. Recently, a number of valuable oxidation or reduction reactions demonstrating the versatility of BESs have been described. Indeed, BESs can produce hydrogen, bring about denitrification, or reductive dehalogenation. Moreover, BESs also appear to be promising in the field of online biosensors. To effectively apply BESs in practice, both biological and electrochemical losses need to be further minimized. At present, the costs of reactor materials have to be decreased, and the volumetric biocatalyst activity in the systems has to be increased substantially. Furthermore, both the ohmic cell resistance and the pH gradients need to be minimized. In this review, these losses and constraints are discussed from an electrochemical viewpoint. Finally, an overview of potential applications and innovative research lines is given for BESs. PMID- 18506437 TI - Clinical pharmacology and pharmacogenetics of thiopurines. AB - The thiopurine drugs-azathioprine (AZA), 6-mercaptopurine (6-MP), and thioguanine are widely used to treat malignancies, rheumatic diseases, dermatologic conditions, inflammatory bowel disease, and solid organ transplant rejection. However, thiopurine drugs have a relatively narrow therapeutic index and are capable of causing life-threatening toxicity, most often myelosuppression. Thiopurine S-methyltransferase (TPMT; EC 2.1.1.67), an enzyme that catalyzes S methylation of these drugs, exhibits a genetic polymorphism in 10% of Caucasians, with 1/300 individuals having complete deficiency. Patients with intermediate or deficient TPMT activity are at risk for excessive toxicity after receiving standard doses of thiopurine medications. This report reviews the recent advances in the knowledge of the mechanism of action as well as the molecular basis and interethnic variations of TPMT and inosine triphosphate pyrophosphatase (ITPase; EC 3.6.1.19), another enzyme implicated in thiopurine toxicity. In addition, an update on pharmacokinetics, metabolism, drug-drug interactions, safety, and tolerability of thiopurine drugs is provided. PMID- 18506440 TI - Effect of the osmotic conditions during sporulation on the subsequent resistance of bacterial spores. AB - The causes of Bacillus spore resistance remain unclear. Many structures including a highly compact envelope, low hydration of the protoplast, high concentrations of Ca-chelated dipicolinic acid, and the presence of small acid-soluble spore proteins seem to contribute to resistance. To evaluate the role of internal protoplast composition and hydration, spores of Bacillus subtilis were produced at different osmotic pressures corresponding to water activities of 0.993 (standard), 0.970, and 0.950, using the two depressors (glycerol or NaCl). Sporulation of Bacillus subtilis was slower and reduced in quantity when the water activity was low, taking 4, 10, and 17 days for 0.993, 0.970, and 0.950 water activity, respectively. The spores produced at lower water activity were smaller and could germinate on agar medium at lower water activity than on standard spores. They were also more sensitive to heat (97 degrees C for 5-60 min) than the standard spores but their resistance to high hydrostatic pressure (350 MPa at 40 degrees C for 20 min to 4 h) was not altered. Our results showed that the water activity of the sporulation medium significantly affects spore properties including size, germination capacity, and resistance to heat but has no role in bacterial spore resistance to high hydrostatic pressure. PMID- 18506441 TI - Occurrence, production, and applications of gellan: current state and perspectives. AB - Bacterial exopolysaccharides (EPS) are products of biotechnology that are of high interest due to their rheological properties. This is the case of sphingans, a group of structurally related EPS secreted by members of the genus Sphingomonas. Among these, gellan is a multifunctional gelling agent produced in high yields by the non-pathogenic strain Sphingomonas elodea ATCC 31461. In its native form, gellan is a linear anionic EPS based on a tetrasaccharide repeat unit composed of two molecules of D: -glucose, one of L: -rhamnose and one of D: -glucuronic acid. The native gellan is partially esterified with acyl substituents (1 mol of glycerate and 0.5 mol of acetate) per repeat unit. Gellan has unique characteristics and has many applications, particularly in the food, pharmaceutical, and biomedical fields. This review summarizes current knowledge on the structure and properties of gellan and provides details about the biosynthesis of this exopolysaccharide. In addition, a highlight of the importance of gellan in industrial and medicinal applications is given. PMID- 18506442 TI - Functional outcome after lumbar closing wedge osteotomy in ankylosing spondylitis. AB - This study entails a prospective evaluation of lumbar closing wedge osteotomy for correction of thoracolumbar kyphotic deformity in ankylosing spondylitis. Twenty patients with a median age of 52 years (range, 26-70) underwent follow-up at one year. The lumbar closing wedge osteomtomy was stabilised by metallic rods fixed by transpedicular screws. Outcome measures were quality of life (EuroQol), occiput-to-wall distance, pain, fatigue, complications, technical and radiological evaluation. The technical result was good in 16 and fair in four patients; two had neuropraxia. The deformity was reduced an average of 17 degrees (95% confidence interval 15-25 degrees) at one-year follow-up. Pain during activity, pain at night, and fatigue were significantly reduced. EuroQol improved from 0.42 to 0.69 (p = 0.002) and occiput-to-wall distance from 26 to 18 cm (p = 0.005). Functional outcome was improved after lumbar closing wedge osteotomy in ankylosing spondylitis. PMID- 18506443 TI - Evaluation of accuracy of an electromagnetic computer-assisted navigation system in total knee arthroplasty. AB - The objective of this study was to develop a method to assess the accuracy of an electromagnetic technology image-free navigation system for total knee arthroplasty in a leg with normal or abnormal mechanical alignment. An acrylic phantom leg was constructed to simulate tibia and femur deformation. Determination of actual leg alignment was achieved using a digital caliper unit. In the setting of normal alignment, the mean error of the system characterised as the difference between the measured computer navigation and digital caliper angles ranged between 0.8 degrees (frontal plane) and 1.5 degrees (lateral plane). In the setting of abnormal alignment, the mean error ranged between 0.4 degrees (frontal plane) and 1.6 degrees (lateral plane). Deformity had no demonstrable effect on accuracy. The study demonstrates satisfactory in vitro system accuracy in both normal and abnormal leg mechanical alignment settings. PMID- 18506444 TI - Progressive subsidence of a tapered, proximally coated femoral stem in total hip arthroplasty. AB - The purpose of this study was to determine if subsidence of a tapered, proximally porous- and hydroxyapatite-coated femoral implant with immediate postoperative weight-bearing differed between men and women. Modified Harris Hip Pain and Function Scores were collected preoperatively, and six weeks and one year postoperatively. Femoral subsidence was determined at six weeks and one year after surgery. Men demonstrated significantly increased Harris Hip Function Scores at six weeks, with slightly inferior Harris Hip Pain Scores at one year. Regardless of sex, subsidence was greater at one year than at six weeks; however, men subsided significantly more at one year than women (1.7 +/- 2.0 mm vs 1.0 +/- 1.4 mm, p = 0.03). While it appears that this stem provided excellent early fixation in women, it is unclear if the increased subsidence in men was more related to patient-related factors of age, weight and activity level or with specific implant characteristics. PMID- 18506445 TI - Accuracy of routine magnetic resonance imaging in meniscal and ligamentous injuries of the knee: comparison with arthroscopy. AB - The aim of this study was to detect the accuracy of routine magnetic resonance imaging (MRI) done in different centres and its agreement with arthroscopy in meniscal and ligamentous injuries of the knee. We prospectively examined 70 patients ranging in age between 22 and 59 years. History taking, plain X-ray, clinical examination, routine MRI and arthroscopy were done for all patients. Sensitivity, specificity, accuracy, positive and negative predictive values, P value and kappa agreement measures were calculated. We found a sensitivity of 47 and 100%, specificity of 95 and 75% and accuracy of 73 and 78.5%, respectively, for the medial and lateral meniscus. A sensitivity of 77.8%, specificity of 100% and accuracy of 94% was noted for the anterior cruciate ligament (ACL). We found good kappa agreements (0.43 and 0.45) for both menisci and excellent agreement (0.84) for the ACL. MRI shows high accuracy and should be used as the primary diagnostic tool for selection of candidates for arthroscopy. LEVEL OF EVIDENCE: 4. PMID- 18506446 TI - Local application of low-dose Depo-Medrol is effective in reducing immediate postoperative back pain. AB - This prospective, randomised case-control study was made to determine effectiveness of low-dose Depo-Medrol applied to the affected nerve root after discectomy. Fifty seven patients with L4-5 or L5-S1 single level disc herniation with unilateral leg pain were selected for the study and were divided in two groups. Twenty eight patients were in the control group and 29 in the steroid group. Discectomy was done after flavotomy in all patients. In the steroid group low-dose 40 mg Depo-Medrol soaked Gelfoam was applied over the affected nerve root after discectomy while in the control group neither saline nor plain Gelfoam was applied to affected root. Postoperatively, patients were asked to evaluate backache using VAS which was compared statistically using the unpaired t test. Statistical difference was significant (p < 0.0001) regarding postoperative VAS during the first month and then it became insignificant. Results show that local application of low-dose Depo-Medrol is helpful in reducing immediate postoperative backache after discectomy, but it is not effective in the long term. PMID- 18506447 TI - [Autopsy records in Cologne from 1914 to 1960: changes in diseases, society and institution]. AB - The first half of the twentieth century was marked by noticeable changes in society, medicine and institutions. The interrelationship between these factors can be demonstrated by clinical mass sources, in particular by patient and autopsy records, which historians have so far neglected. A longitudinal study was made where data, such as gender and age as well as clinical and anatomical pathological diagnoses were collected, based on 250 autopsy records from 8 selected years between 1914 and 1960. The random samples taken showed clear differences in gender relationship and in age composition and in addition they revealed a significant reduction in mortality caused by inflammation and an increase of chronic diseases, such as tumors or cardiac-circulation ailments. The origin of this development is seen in medical progress as well as in institutional and ideological circumstances, therefore multidimensional reciprocal effects must be thoroughly analyzed. PMID- 18506448 TI - The single nucleotide polymorphisms of matrix metalloproteinase-1 in patients with systemic sclerosis. PMID- 18506449 TI - Cell tracking with optical imaging. AB - Adaptability, sensitivity, resolution and non-invasiveness are the attributes that have contributed to the longstanding use of light as an investigational tool and form the basis of optical imaging (OI). OI, which encompasses numerous techniques and methods, is rapid (<5 min), inexpensive, noninvasive, nontoxic (no radiation) and has molecular (single-cell) sensitivity, which is equal to that of conventional nuclear imaging and several orders of magnitude greater than MRI. This article provides a comprehensive overview of emerging applications of OI based techniques for in vivo monitoring of new stem cell-based therapies. Different fluorochromes for cell labeling, labeling methods and OI-based cell tracking techniques will be reviewed with respect to their technical principles, current applications and aims for clinical translation. Advantages and limitations of these new OI-based cell-tracking techniques will be discussed. Non invasive mapping of cells labeled with fluorochromes or OI marker genes has the potential to evolve further within the clinical realm. PMID- 18506450 TI - Bone scintigraphy is really unnecessary for evaluation of bone metastasis? PMID- 18506451 TI - The cold but not hard fats in ectotherms: consequences of lipid restructuring on susceptibility of biological membranes to peroxidation, a review. AB - The production of reactive oxygen species is a regular feature of life in the presence of oxygen. Some reactive oxygen species possess sufficient energy to initiate lipid peroxidation in biological membranes, self-propagating reactions with the potential to damage membranes by altering their physical properties and ultimately their function. Two of the most prominent patterns of lipid restructuring in membranes of ectotherms involve contents of polyunsaturated fatty acids and ratios of the abundant phospholipids, phosphatidylcholine and phosphatidylethanolamine. Since polyunsaturated fatty acids and phosphatidylethanolamine are particularly vulnerable to oxidation, it is likely that higher contents of these lipids at low body temperature elevate the inherent susceptibility of membranes to lipid peroxidation. Although membranes from animals living at low body temperatures may be more prone to oxidation, the generation of reactive oxygen species and lipid peroxidation are sensitive to temperature. These scenarios raise the possibility that membrane susceptibility to lipid peroxidation is conserved at physiological temperatures. Reduced levels of polyunsaturated fatty acids and phosphatidylethanolamine may protect membranes at warm temperatures from deleterious oxidations when rates of reactive oxygen species production and lipid peroxidation are relatively high. At low temperatures, enhanced susceptibility may ensure sufficient lipid peroxidation for cellular processes that require lipid oxidation products. PMID- 18506452 TI - Sign on the dotted line: parental consent. AB - The issue of informed consent in paediatric surgery has always been contentious. Despite the fact that taking consent is one of the most frequently performed tasks of a surgeon, it is rarely audited. Indeed, there are few studies looking at the consent process in adult practice and fewer in the paediatric setting. The aim of this study was, therefore, to determine parental understanding, recall, attitudes and opinion of the consent process in a busy paediatric day surgery unit. A prospective study was conducted using a questionnaire consisting of 20 questions. These were handed out to the parents of all children undergoing one of four procedures: circumcision, prepuceplasty, orchidopexy and herniotomy over a 3 month-period. Completed questionnaires were collected and analysed. One hundred and sixty-one parents were asked to complete the questionnaire. Ninety-three were collected, a response rate of 58%, 87% believed consent was taken by the performing surgeon, 15% felt the consent process was rushed, but 91% thought the right amount of detail was given. Ninety-seven percent of parents believed that the consent form was a legal necessity and 72% believed that a child in Scotland had to be aged 16 or over to sign the consent form. Interestingly, 23% of parents were unsure if signing the consent form meant that they could not claim compensation if anything went wrong. Ninety-one percent recalled the procedure being explained in the outpatient clinic, but 22% did not remember potential complications being discussed. The amount parents recalled of potential complications varied. Seventy percent felt that a leaflet about the procedure prior to attending the unit would be helpful. In conclusion consent is an extremely important part of our practice. This study highlights some of the common parental misconceptions and limitations of the process. Parents appear satisfied and consenting for procedures is overall performed well. Some areas could be improved and more research is required in this area. PMID- 18506453 TI - Clinical characteristics and quality of life in a cohort of 621 patients with faecal incontinence. AB - AIM: The aim of this study was to study a cohort of patients with faecal incontinence (FI) to gain a better insight into the clinical and epidemiological characteristics of this pathology and its repercussions on quality of life (QL). MATERIALS AND METHODS: Consecutive patients with FI seen at tertiary centres filled in a self-questionnaire. The severity of FI, constipation and urinary incontinence (UI) was evaluated, respectively, by the Jorge and Wexner score, the Knowles-Eccersley-Scott Symptom score and the Urological Distress Inventory score. ROME II criteria were used to assess the existence of an associated irritable bowel syndrome. The repercussion on QL was evaluated by the Gastrointestinal Quality of Life index score and the Ditrovie score. The psychological status was assessed by the Hospital Anxiety and Depression scale. RESULTS: Six hundred twenty-one patients (114 men), mean age 58 +/- 15 years (range: 20-92), with FI, filled in the questionnaire. The mean Jorge and Wexner score was 11 +/- 4. Twenty-seven presented with an irritable bowel syndrome. Thirty-eight percent had an associated constipation. A UI was associated in 48% women and 25% men. QL was significantly altered, and anxiety and depression were frequent. CONCLUSIONS: FI symptoms are frequently severe, QL very altered and anxiety and depression common. FI is frequently associated with other digestive and perineal symptoms, which argue in favour of a multi-disciplinary management of FI. PMID- 18506454 TI - Meta-analysis of different test indicators: Helicobacter pylori infection and the risk of colorectal cancer. AB - BACKGROUND AND AIMS: Recent studies have demonstrated the relationship between Helicobacter pylori infection and the risk of colorectal carcinoma. However, the results of these studies remain controversial as the studies were relatively small in size and partially differed in designs, and so we reviewed the published studies and carried out a meta-analysis to further explore this relationship. MATERIALS AND METHODS: We performed an extensive systematic review to find all the published case-control studies up to Jan. 2007 using electronic searching, hand searching, and reference lists of retrieved articles. Odds ratio (OR) was employed to evaluate the relationship of H. pylori infection and risk of colorectal cancer. Summary estimates were obtained using random effect models according to the result of a statistical test for heterogeneity across the studies. The presence of possible publication bias was assessed using different statistical approaches. RESULTS: Thirteen studies were included, and summary OR 1.49 (95% confidence interval [CI] 1.17-1.91) was estimated for the association between H. pylori infection and colorectal cancer. Summary OR 1.56 (95% CI 1.14 2.14) was estimated for the association between immunoglobulin G antibody and colorectal cancer risk. By trimming and filling, the number of inputted studies was zero, and summary OR was still 1.49 (95% CI 1.17-1.91). The graphical funnel plot appeared asymmetrical, but there was no statistical evidence of publication bias. The method of fail-safe suggested that the effect of publication bias was small. CONCLUSION: Current evidence, though limited, suggests that there is a possible increase in risk of colorectal cancer because of H. pylori infection. PMID- 18506456 TI - Technetium-99 m-MDP scintigraphy and long-term follow-up of musculo-skeletal sarcoma reconstructed with pasteurized autologous bone graft. AB - INTRODUCTION: Local bone tumor excision followed by pasteurization and reimplantation is a unique technique of reconstruction for sarcoma patients. The long-term scintigraphic findings of pasteurized bone grafts in relation to clinical patients' data were reviewed retrospectively. PATIENTS: Twenty-two sarcoma patients (mean age, 24 years) done between April 2000 and January 2004 constituted the material of this study. One hundred and fifty-two 99 m TC methylene diphosphonate (MDP) whole-body scans were reviewed. Initially, all autografts appeared as photon deficient areas. Diffusely increased bone uptake was present at the osteotomy sites within 4-6 months after surgery, the uptake of the grafted pasteurized bone was detected in 17 patients from about 6 months after surgery. RESULTS: Of 22 patients, 11 (50%) showed higher uptake than the normal bone, 6 (27%) had uptake similar to the normal bone, while 5 (23%) had less uptake than the normal bone. Radiologically, 15 patients (68%) showed complete incorporation of graft and 5 patients (23%) had partial incorporation. Oncologically, 16 patients are disease free, while 6 died of disease. No local recurrence was detected at a mean of 59 months, while fracture (13.6%) eventually healed with bone grafting and revision of internal fixation, graft collapse (9%) (needed revision arthroplasty) and infection (9%), one cured and one converted to rotationplasty, were the major complications encountered. Significantly more mechanical complications were seen when tracer uptake suggestive of revascularization occurred; thus, revascularization and partial bone ingrowth are not sufficient conditions for lower mechanical complication rate. CONCLUSION: The method of pasteurization of bone is a useful option for reconstruction after resection of malignant bone tumors. PMID- 18506455 TI - TDP-43 protein in plasma may index TDP-43 brain pathology in Alzheimer's disease and frontotemporal lobar degeneration. AB - Autopsy studies have shown that about 55% of patients with frontotemporal lobar degeneration (FTLD) and 25% of patients with Alzheimer's disease (AD) harbour TDP 43 immunoreactive pathological changes in their brains. Using ELISA, we investigated whether we could detect the presence, or increased amounts, of TDP 43 in plasma of patients with FTLD and AD compared to normal control subjects. We detected elevated levels of TDP-43 protein in plasma of 46% patients with FTLD with clinical frontotemporal dementia (FTD) and 22% patients with AD, compared to 8% of control subjects. The proportions of patients with FTD and AD showing raised plasma TDP-43 levels correspond closely to those proportions known from autopsy studies to contain TDP-43 pathological changes in their brains. Raised TDP-43 plasma levels may thereby index TDP-43 pathology within the brain. Plasma TDP-43 levels may be a biomarker that can provide a laboratory test capable of identifying the presence of TDP-43 brain pathology in neurodegenerative disease during life. It may help to distinguish those cases of FTLD with ubiquitin/TDP-43 pathology in their brains from those with tauopathy. As a predictive test, plasma TDP-43 level may have great practical value in directing therapeutic strategies aimed at preventing or removing tau or TDP-43 pathological changes from the brain in FTLD and AD. PMID- 18506457 TI - Skin graft failure mimicking abscess masking local recurrence of vulvar cancer: a case report. AB - BACKGROUND: Vulvar cancer is a rare gynecological malignancy and its local recurrence is even more uncommon. To date, no cases of recurrent vulvar cancer in split thickness graft area have been reported. We describe the first case of such recurrence. CASE: A 65-year-old G4P3013 presented with vulvar mass and itching. She had a 2 x 2-cm sized mass on clitoris and an ulcerated bulgy mass of 2 x 2.5 cm size was localized on left labia majora, of which a biopsy revealed invasive squamous cell carcinoma. She underwent a radical vulvectomy and bilateral inguinofemoral lymph node dissection via separate incisions. Her postoperative course was complicated by wound separation which was cored with a split thickness skin graft. Patient was advised to integrate her treatment with radiotherapy, but she refused it. Two months later, she consulted for a painful swelling in the operation field. The swelling resembled an abscess formation so it was drained and an outpatient antibiotic treatment was prescribed. At her second visit, it was noted that the above-mentioned lesion was persisting and so fine needle aspiration biopsy (FNAB) was performed and a local recurrence was retrieved beneath the graft failure. Then she underwent a wide local excision and adjuvant radiotherapy was initiated after patient's approval. CONCLUSION: Careful surveillance is warranted as squamous cell carcinoma recurrence can even occur beneath the split thickness skin graft, especially in conjunction with graft failure. And for high risk patients adjuvant radiotherapy is critically important for avoiding recurrences. PMID- 18506458 TI - Tumor markers in mature cystic teratomas of the ovary. AB - OBJECTIVE: The objective of this study was to evaluate the serum levels of tumor markers in patients with ovarian mature cystic teratomas. METHOD: Retrospective study of 215 patients operated at Zekai Tahir Burak Women Health Education and Research Hospital between January 2001 and October 2006 was performed. RESULTS: The median age was 36 years (range 13-80). The mean tumor diameter was 7.7 +/- 4.6 cm (range 2-25). The mean serum CA 125 level was 26.2 +/- 29.9 U/mL (range 1.4-225, normal value <35), and the mean serum CA 19-9 level was 83.5 +/- 179.2 IU/mL (range 0.6-1,000; normal value <37). The elevated rate of CA 19-9, CA 125 was 39.6% (74/187) and 23.3% (50/215), respectively. The mean age of patients and the rate of bilaterality of tumors were similar in both patients with elevated CA19-9 and with normal CA 19-9 level (P > 0.05). The mean tumor size of patients with elevated CA 19-9 was greater than those with normal CA19-9 level (P = 0.01). CONCLUSION: Serum CA 19-9 has the highest positivity rate among other tumor markers in ovarian mature cystic teratomas. Elevated serum CA 19-9 levels are correlated with larger tumor size. But the diagnostic value of elevated CA 19-9 in patients with MCT would be poor if the test was used alone. PMID- 18506459 TI - Pemphigoid gestationis: a rare case and review. AB - Pemphigoid gestationis is a rare, autoimmune blistering dermatosis (Holmes and Black In: Clin Exp Dermatol 7:65-73, 1982). It is unique in that it is diagnosed primarily in association with pregnancy. It has been estimated to occur in 1:50,000 pregnancies [Shornick et al. In: J Am Acad Dermatol 8:214-224, 1983]. Involvement of the infant is exceedingly rare, while the pathogenic trigger initiating disease has remained elusive; the interaction of IgG1 auto-antibodies and the 180 kD BPA2 remains paramount in understanding the immunologic mechanism of disease. We describe a 29 year-old multiparous woman who presented at 29 weeks gestation with skin findings characteristic of pemphigoid gestationis. She later gave birth to a full-term male who was born with extensive bullae and several denuded areas of his head, trunk and limbs. The diagnosis of pemphigoid gestationis in both mother and son was confirmed using direct immunofluorescence. In conclusion, we present a well documented, rare case of pemphigoid gestationis with maternal transfer to the infant. A historical and epidemiological review of this rare blistering disorder is discussed along with current theories on pathogenic and immunologic mechanisms. PMID- 18506460 TI - Retained foetal bones: an intrauterine cause of chronic pelvic pain. AB - Intrauterine retention of foetal bones is an uncommon but recognised complication of late termination of pregnancy. Secondary subfertility, abnormal uterine bleeding and vaginal discharge are the usual presenting complaints. We report a case of prolonged retention of foetal bones for 14 years in a woman who presented with chronic pelvic pain. Hysteroscopic examination was diagnostic and therapeutic. Retained foetal bones are an uncommon intrauterine cause of chronic pelvic pain that should be considered particularly when a woman with a history of late termination presents with pelvic pain. Hysteroscopic evacuation is curative. PMID- 18506461 TI - Outcome of induced deliveries in growth-restricted fetuses: second thoughts about the vaginal option. AB - OBJECTIVE: To assess the outcome of induced deliveries with IUGR. STUDY DESIGN: We reviewed the computerized files of parturients who underwent inducted labor because of IUGR (<5th percentile). Outcome assessment included mode of delivery, indication for CS, NICU admissions and 5 min Apgar score. We compared these parameters to a control group of women whose deliveries were induced for other indications. RESULTS: A total of 836 women with IUGR were included. Mean gestational age and birth weight were 38.2 weeks and 2,114 g, respectively. Overall, 43% of women delivered by non-elective C/S. The rate of non-elective CS for other indications was 12.3% (P < 0.0001) for all deliveries and 23.6% (P < 0.001) for induced deliveries. CS was performed due to non-reassuring FHR in 63% of IUGR fetuses, compared with 27% of all induced deliveries. There were 160 women with IUGR who preferred elective CS. Their newborns' NICU admission and 5 min Apgar score <7 rates were lower than those for induced deliveries (NICU 43.1 and 29.4%, P < 0.05, 5 min Apgar <7 5 and 1%, P < 0.05). CONCLUSION: Growth restricted neonates born after labor induction had higher rates of low Apgar scores and NICU admissions compared to growth restricted neonates delivered by elective C/S. Inductions of labor for IUGR were associated with higher rates of non-elective C/S due to non-reassuring fetal heart rate compared with inductions performed for other indications. PMID- 18506462 TI - Uterocutaneous fistula after surgical treatment of an incomplete abortion: methylene blue test to verify the diagnosis. AB - INTRODUCTION: Uterocutaneous fistula is an extremely rare clinical condition that can be seen after pelvic or uterine surgery. It can also complicate some obstetric procedures. CASE: We report of an unusual case of an uterocutaneous fistula that developed in a multiparous woman after surgical evacuation of an incomplete first trimester septic abortion. The fistula tract was depicted on computed tomography, and to verify the diagnosis methylene blue was given through a transcervically introduced uterine catheter, and blue dye flow out through the external opening of fistula was observed. At laparatomy fistula tract was completely excised along with the enclosing omentum. Postoperative recovery and follow-up were uneventful. DISCUSSION: Possible mechanisms of development of such a rare condition, and diagnostic and treatment options are discussed. PMID- 18506463 TI - Acute kidney injury requiring dialysis in obstetric patients: a series of 55 cases in Brazil. AB - OBJECTIVE: To investigate the risk factors associated with oliguria and death in obstetric patients with acute kidney injury (AKI). METHODS: The study group included all obstetric patients with AKI, under dialytic treatment, in Hospital Geral Cesar Cals, Fortaleza, Brazil, from January 2000 to December 2006. AKI were classified according to the RIFLE criteria. Univariate and multivariate analysis were performed to investigate the factors associated with oliguria and death. RESULTS: A total of 55 patients were included. Their average age was 26.2 +/- 6.7 years. The main etiologies of AKI were pregnancy-related hypertension (41.8%), HELLP syndrome (40%), puerperal sepsis (14.5%), abruption placentae (9.1%), hemolytic uremic syndrome (9.1%) and thrombotic thrombocytopenic purpura (5.5%). Oliguria was observed in 36 cases (65%). Death occurred in 17 cases (30.9%). Factors associated with oliguria were, diagnosis of HELLP syndrome, hyperbilirubinemia and death. Factors associated with death were, presence of puerperal sepsis, hyperbilirubinemia, hypotension, oliguria and low levels of HCO(3). CONCLUSION: AKI is a rare but potential fatal complication in obstetric patients. RIFLE criteria seem to have association with mortality. There are important factors associated with oliguria and death, which must be prompt recognized to the institution of adequate therapeutic measures. PMID- 18506464 TI - Relationship between clinical diagnosis of recurrent vulvovaginal candidiasis and detection of Candida species by culture and polymerase chain reaction. AB - BACKGROUND: Recurring vulvovaginal candidiasis (RVVC) is a common vaginal discharge affecting 75% of all women at least once in their life. In 5% of these women, infection is recurring. Aim of the study was to determine the sensitivity of detecting Candida species by culture and polymerase chain reaction (PCR) in women with a clinical diagnosis of RVVC. METHODS: A total number of 104 patients referred with a clinical diagnosis of RVVC and therefore at least four episodes in the previous year were evaluated. In order to detect Candida, vaginal swabs were cultured on Sabouraud and chromagar. Furthermore, the supernatant from the vaginal lavage was examined for the presence of Candida by PCR. RESULTS: When the culture was analyzed, only 31 (29.8%) of the 104 patients diagnosed with RVVC were positive for Candida species in their vagina. Candida albicans was identified in 25 women and six were positive for Candida glabrata. When analyzed by PCR, 44 (42.3%) patients were positive for Candida species. In 13 women (12.5%) only the PCR was positive, while in 31 patients both culture and PCR were positive. CONCLUSION: The diagnostic method of PCR is more sensitive than culture in detecting Candida species in the vagina. The results also suggest further investigation to verify the complaints of the negative tested patients. PMID- 18506465 TI - A simple method to alleviate aspiration in the near-total laryngectomy patient. AB - Near-total laryngectomy is a surgical technique which grants the potential for postoperative speech without the need for prostheses or secondary surgical procedures. Aspiration can be a problem, however, that can require completion laryngectomy to resolve. A 60-year-old male underwent a near-total laryngectomy for recurrent laryngeal cancer. The patient developed chronic aspiration secondary to a leaking shunt. Work up was negative for recurrent cancer. Calcium hydroxylapatite was injected transorally at the opening into the shunt and transtomally into the exit of the shunt to seal it. Postoperative barium swallow showed resolution of aspiration. At approximately 17 months, the patient developed recurrent intermittent aspiration of thin liquids and required reinjection of the shunt, with resolution of the aspiration. Calcium hydroxylapatite allows simple and effective alleviation of aspiration following near-total laryngectomy but requires repeated injection to maintain efficacy. Injection of calcium hydroxylapatite can be an effective alternative to completion laryngectomy in patients who aspirate following near-total laryngectomy. PMID- 18506467 TI - Lethal outcome of post-tonsillectomy hemorrhage. AB - Despite the large number of tonsillectomies performed little knowledge exists about post-tonsillectomy hemorrhage (PTH) with lethal outcome. This study was performed to evaluate clinical features in a larger patient population with emphasis on the onset of this complication. A nationwide collection of cases was performed based on personal communication, expert reports to lawsuits and professional boards, and case reports received after a plea published in a professional national journal. Clinical data of 29 patients were collected of whom the 18 were children (64%). With one exception all patients experienced secondary PTH (>24 h) occurring 1-28 days after tonsillectomy. Aspiration contributed to lethal outcome in 13 cases. Fatalities were unavoidable although 21 patients were in the hospital. Massive vomiting of blood was observed in 11 patients. There were 11 patients without (group A) and 18 with (group B) episodes of repeated bleeding. This study suggests that particularly children are endangered by lethal PTH. Inpatient treatment was unable to prevent lethal outcome in this selected patient population. However, it appears wise to re-admit patients with delayed PTH, since excessive PTH may occur. These unexpected and unpredictable situations require an immediate and adequate medical treatment by a skilled staff. The paucity of data currently does not allow calculation of a cut off point at which the risk of life-threatening PTH significantly decreases. Secondary PTH remains a substantial complication. PMID- 18506466 TI - The value of a liquid alginate suspension (Gaviscon Advance) in the management of laryngopharyngeal reflux. AB - Laryngopharyngeal reflux (LPR) refers to the backflow of stomach contents into the laryngopharynx. Increasing evidence has demonstrated that LPR is a contributing factor in some cases of hoarseness, vocal fatigue, voice breaks, cough and globus and chronic throat clearing. However, several randomised placebo controlled trials of proton pump inhibitors in the treatment of LPR have been reported with the majority showing no significant benefit in patient symptom scores over placebo. The aim of this pilot clinical study was to investigate whether any improvement in LPR-related symptoms, using the Reflux Symptom Index (RSI), and clinical findings, using the Reflux Finding Score (RFS), could be achieved with treatment with a liquid alginate suspension compared to control (no treatment). Patients presenting with the symptoms of LPR to the Otorhinolaryngology Outpatient Department at the Queen's Medical Centre, Nottingham, UK were considered eligible if they had an RSI of greater than 10 and an RFS greater than 5 based on a fibreoptic examination of the larynx. A total of 49 patients were randomised into the open, parallel group study; 24 patients were randomised to receive 10 ml liquid alginate suspension (Gaviscon Advance) four times daily after meals and at bedtime, and 25 patients into the control group (no treatment). Patients were assessed pre-treatment and at 2, 4 and 6 months post treatment. Mean (SD) RSI and RFS pre-treatment scores were 23.9 (7.0) and 10.4 (3.6) for the treatment group and 24.6 (7.4) and 10.3 (3.3) for the control group, respectively. Significant differences between treatment and control were observed for RSI at the 2-month (11.2 (7.0) vs. 16.8 (6.4), P=0.005) and 6-month (11.2 (8.1) vs. 18.3 (9.4), P=0.008) assessments and for RFS at the 6-month (7.1 (2.8) vs. 9.5 (3.4), P=0.005) assessment. Significant improvement in symptom scores and clinical findings were achieved with liquid alginate suspension (Gaviscon Advance) compared to control and further evaluation for the management of patients presenting with LPR is warranted. PMID- 18506468 TI - Necrotizing otitis externa: a painless reminder. AB - Necrotizing otitis externa is a necrotizing osteomyelitis of the skull base that predominantly affects diabetic and immunocompromised patients. Pseudomonas aeruginosa is the most frequently implicated organism. Patients present with severe otalgia, otorrhoea with or without facial palsy, or rarely, facial pain as a symptomatic feature. We report a case of necrotizing otitis externa presenting with totally painless otorrhoea and highlight the need for a high index of suspicion when treating patients with otitis externa. PMID- 18506469 TI - Imaging aspects of cardiovascular disease at the cell and molecular level. AB - Cell and molecular imaging has a long and distinguished history. Erythrocytes were visualized microscopically by van Leeuwenhoek in 1674, and microscope technology has evolved mightily since the first single-lens instruments, and now incorporates many types that do not use photons of light for image formation. The combination of these instruments with preparations stained with histochemical and immunohistochemical markers has revolutionized imaging by allowing the biochemical identification of components at subcellular resolution. The field of cardiovascular disease has benefited greatly from these advances for the characterization of disease etiologies. In this review, we will highlight and summarize the use of microscopy imaging systems, including light microscopy, electron microscopy, confocal scanning laser microscopy, laser scanning cytometry, laser microdissection, and atomic force microscopy in conjunction with a variety of histochemical techniques in studies aimed at understanding mechanisms underlying cardiovascular diseases at the cell and molecular level. PMID- 18506470 TI - Expression and regulation of c-Jun N-terminal kinase (JNK) in endometrial cells in vivo and in vitro. AB - JNK(c-Jun N-terminal kinase) is one of the main types of mitogen-activated protein kinases. JNK modulates inflammation and apoptosis in response to stress. Our hypothesis is that temporal and spatial changes in JNK activity regulate inflammation in human endometrium and that fluctuation in estrogen and progesterone levels may play a role in JNK activation. Therefore, we aimed to determine total-(t-) and active-(phosphorylated, p-) JNK expression in endometrial tissues in vivo by immunohistochemistry, and in vitro by immunocytochemistry and Western blot analysis. Immunohistochemistry revealed moderate cytoplasmic and nuclear t-JNK immunoreactivity, and mostly nuclear p-JNK immunoreactivity throughout the menstrual cycle and early pregnancy. The highest p- and t-JNK immunoreactivity was detected in late secretory phase (P < 0.05). We observed that endometrial stromal cell (ESC)s showed a significant increase in p JNK expression following 48 h of estrogen combined with progesterone (E(2) + P(4)) withdrawal from the culture conditions, compared to control and non withdrawal groups (P < 0.05). Upon treatment with JNK inhibitor SP600125, we observed a significantly decreased interleukin (IL)-8 level (P < 0.05) in the presence and absence of E(2). These results demonstrate that JNK expression increases during the late secretory phase when the inflammatory response is highest. Inhibition of IL-8 expression by SP600125 suggests that JNK is involved in regulation of proinflammatory mediators of endometrium. PMID- 18506471 TI - Work organization and drinking: an epidemiological comparison of two psychosocial work exposure models. AB - OBJECTIVES: To examine the relationship between two alternative psychosocial work exposure measurement frameworks (i.e., job alienation and job stress) and three drinking behaviours (frequent, heavy and drinking and work). METHODS: A cross sectional survey was conducted among 3,099 US drinking workers. Job stress conditions were assessed according to the Karasek's job strain model. Alienating job conditions were assessed with measures based on Kohn and Schooler's occupational self-direction concept. Multivariate logistic regression controlling for a wide range of known covariates was used. RESULTS: High strain work showed no associations, while workers in passive jobs had an increased likelihood of heavy (OR = 1.29; 95%CI: 1.02-1.64) and lower likelihood of frequent drinking (OR = 0.71; 95%CI: 0.52-0.97). Unexpectedly, low complexity combined with low constraint related to more frequent drinking (OR = 1.60; 95%CI: 1.22-2.10). No associations with drinking at work were observed. CONCLUSION: Our findings suggest an association between different work environment features and drinking behaviours. Our findings highlight the value of exploring the hypothesized passive pathway of the job strain model together with other theoretical perspectives, such as alienating job conditions. PMID- 18506472 TI - Postural destabilization induced by trunk extensor muscles fatigue is suppressed by use of a plantar pressure-based electro-tactile biofeedback. AB - Separate studies have reported that postural control during quiet standing could be (1) impaired with muscle fatigue localized at the lower back, and (2) improved through the use of plantar pressure-based electro-tactile biofeedback, under normal neuromuscular state. The aim of this experiment was to investigate whether this biofeedback could reduce postural destabilization induced by trunk extensor muscles. Ten healthy adults were asked to stand as immobile as possible in four experimental conditions: (1) no fatigue/no biofeedback, (2) no fatigue/biofeedback, (3) fatigue/no biofeedback and (4) fatigue/biofeedback. Muscular fatigue was achieved by performing trunk repetitive extensions until maximal exhaustion. The underlying principle of the biofeedback consisted of providing supplementary information related to foot sole pressure distribution through electro-tactile stimulation of the tongue. Centre of foot pressure (CoP) displacements were recorded using a force platform. Results showed (1) increased CoP displacements along the antero-posterior axis in the fatigue than no fatigue condition in the absence of biofeedback and (2) no significant difference between the no fatigue and fatigue conditions in the presence of biofeedback. This suggests that subjects were able to efficiently integrate an artificial plantar pressure information delivered through electro-tactile stimulation of the tongue that allowed them to suppress the destabilizing effect induced by trunk extensor muscles fatigue. PMID- 18506473 TI - Partial thyroidectomy under local anaesthesia-the analysis of 49 subsequent cases. AB - BACKGROUND AND AIMS: Partial thyroidectomy under local anaesthesia was performed in 49 subsequent individuals in the Central African Republic. Because of the shortage of medical resources, all patients with goitre were scheduled for an operation under local anaesthesia. MATERIALS AND METHODS: There were no inclusion or exclusion criteria applied for patient selection for the procedure. Before the operation, patients had received an oral sedation and antibiotic. For infiltration anaesthesia, 1% lignocaine was used. RESULTS: Subtotal bilateral thyroidectomy was performed in 37 patients; 12 patients underwent lobectomy or partial lobectomy of the affected portion of the gland. There were no intraoperative complications in any of the patients. The mean time of the procedure averaged 127 min. There were no postoperative complications noticed in the reported group, and this includes also complications related to laryngeal nerve injury. General condition of the operated on patients allowed for full self dependency within 4 to 6 h postoperatively. Patients remained under medical surveillance for mean 3 days, and cutaneous stitches were removed on the first postoperative day. General condition of all patients on the day of discharge from hospital was good. CONCLUSION: Surgery for goitre under local anaesthesia may be a safe alternative where general anaesthesia is not available or contraindicated for medical reasons. PMID- 18506474 TI - "Minimally invasive video-assisted thyroidectomy. Initial experience in a general surgery department". AB - BACKGROUND AND AIMS: The aim of this study is to analyze our preliminary results from minimally invasive video-assisted thyroidectomy (MIVAT) and demonstrate the feasibility of MIVAT also in non-referral centers. MATERIALS AND METHODS: We report our initial experience based on a series of 47 patients selected for MIVAT at General Surgery Department of University of Trieste during a period from May 2005 to February 2007. The eligibility criteria were rigorously observed. Age, goiter volume, major diameter of the dominant nodule, operative times, pathologic findings, postoperative pain, length of hospital stay, cosmetic results, and complications were retrospectively analyzed. RESULTS: Thyroid lobectomy was successfully accomplished in 33 cases, total thyroidectomy in 14. Conversion to standard cervicotomy was required in three patients (6%). Mean operative time of lobectomy was 82.6 min and 118.7 for total thyroidectomy. Postoperative complications included 11 (23.4%) transient hypocalcemias, 2 (4.2%) hematomas, and 2 (4.2%) temporary laryngeal nerve palsies. None-recurrent nerve palsies was observed. The cosmetic result was excellent in most cases. CONCLUSIONS: Our experience demonstrates that MIVAT, after adequate training, is feasible and safe, with results comparable to conventional thyroidectomy, also in a General Surgery Department, from a dedicated team, with a sufficient and specific activity volume. PMID- 18506475 TI - Ascorbic acid participates in a general mechanism for concerted glucose transport inhibition and lactate transport stimulation. AB - In this paper, we present a novel function for ascorbic acid. Ascorbic acid is an important water-soluble antioxidant and cofactor in various enzyme systems. We have previously demonstrated that an increase in neuronal intracellular ascorbic acid is able to inhibit glucose transport in cortical and hippocampal neurons. Because of the presence of sodium-dependent vitamin C transporters, ascorbic acid is highly concentrated in brain, testis, lung, and adrenal glands. In this work, we explored how ascorbic acid affects glucose and lactate uptake in neuronal and non-neuronal cells. Using immunofluorescence and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, the expression of glucose and ascorbic acid transporters in non-neuronal cells was studied. Like neurons, HEK293 cells expressed GLUT1, GLUT3, and SVCT2. With radioisotope-based methods, only intracellular ascorbic acid, but not extracellular, inhibits 2-deoxyglucose transport in HEK293 cells. As monocarboxylates such as pyruvate and lactate, are important metabolic sources, we analyzed the ascorbic acid effect on lactate transport in cultured neurons and HEK293 cells. Intracellular ascorbic acid was able to stimulate lactate transport in both cell types. Extracellular ascorbic acid did not affect this transport. Our data show that ascorbic acid inhibits glucose transport and stimulates lactate transport in neuronal and non-neuronal cells. Mammalian cells frequently present functional glucose and monocarboxylate transporters, and we describe here a general effect in which ascorbic acid functions like a glucose/monocarboxylate uptake switch in tissues expressing ascorbic acid transporters. PMID- 18506476 TI - TRESK-like potassium channels in leukemic T cells. AB - In this study, we present patch-clamp characterization of the background potassium current in human lymphoma (Jurkat cells), generated by voltage independent 16 pS channels with a high ( approximately 100-fold) K+/Na+ selectivity. Depending on the background K+ channels density, from few per cell up to approximately 1 open channel per microm2, resting membrane potential was in the range of -40 to -83 mV, approaching E (K) = -88 mV. The background K+ channels were insensitive to margotoxin (3 nM), apamine (3 nM), and clotrimazole (1 microM), high-affinity blockers of the lymphocyte Kv1.3, SKCa2, and IKCa1 channels. The current depended weakly on external pH. Arachidonic acid (20 microM) and Hg2+ (0.3-10 microM) suppressed background K+ current in Jurkat cells by 75-90%. Background K+ current was weakly sensitive to TEA+ (IC50 = 14 mM), and was efficiently suppressed by externally applied bupivacaine (IC50 = 5 microM), quinine (IC50 = 16 microM), and Ba2+ (2 mM). Our data, in particular strong inhibition by mercuric ions, suggest that background K+ currents expressed in Jurkat cells are mediated by TWIK-related spinal cord K+ (TRESK) channels belonging to the double-pore domain K+ channel family. The presence of human TRESK in the membrane protein fraction was confirmed by Western blot analysis. PMID- 18506477 TI - Stamen development in Arabidopsis is arrested by organ-specific overexpression of a cucumber ethylene synthesis gene CsACO2. AB - Cucumber (Cucumis sativus L.) has served as a model to understand hormone regulation in unisexual flower development since the 1950s and the role of ethylene in promoting female flower development has been well documented. Recent studies cloned the F-locus in gynoecious lines as an additional copy of the ACC synthase (ACS) gene, which further confirmed the role of ethylene in the promotion of female cucumber flowers. However, no direct evidence was generated to demonstrate that increases in endogenous ethylene production could induce female flowers by arresting stamen development. To clarify the relationship between ethylene production and stamen development, we overexpressed the ethylene synthesis cucumber gene CsACO2 to generate transgenic Arabidopsis, driven by the organ-specific promoter P ( AP3 ). We found that organ-specific overexpression of CsACO2 significantly affected stamen but not carpel development, similar to that in the female flowers of cucumber. Our results suggested that increases in ethylene production directly disturb stamen development. Additionally, our study revealed that among all floral organs, stamens respond most sensitively to exogenous ethylene. PMID- 18506478 TI - Molecular cloning, functional characterization and expression analysis of a novel monosaccharide transporter gene OsMST6 from rice (Oryza sativa L.). AB - Monosaccharides transporters play important roles in assimilate supply for sink tissue development. In this study, a new monosaccharide transporter gene OsMST6 was identified from rice (Oryza sativa L.). The predicted OsMST6 protein shows typical features of sugar transporters and shares 79.6% identity with the rice monosaccharide transporter OsMST3. Heterologous expression in yeast (Saccharomyces cerevisiae) demonstrated that OsMST6 is a broad-spectrum monosaccharide transporter, with a K (m) of 266.1 muMu for glucose. OsMST6-green fluorescent protein fusion protein is localized to the plasma membrane in plant. Semi-quantitative RT-PCR analysis exhibited that OsMST6 is expressed in all tested organs/tissues. In developing seeds, OsMST6 expression level is high at the early and middle grain filling stages and gradually declines later. Further analysis detected its expression in both maternal and filial tissues. RNA in situ hybridization analysis indicated that OsMST6 is predominantly expressed in the vascular parenchyma of the chalazal vein, cross-cells, nucellar tissue and endosperm of young seeds, in mesophyll cells of source leaf blades, and in pollens and the connective vein of anthers. In addition, OsMST6 expression is up regulated by salt stress and sugars. The physiological role of OsMST6 for seed development and its roles in other sink and source tissues are discussed. PMID- 18506479 TI - PRD, an Arabidopsis AINTEGUMENTA-like gene, is involved in root architectural changes in response to phosphate starvation. AB - Changes in root architecture are one of the adaptive strategies used by plants to compensate for local phosphate (Pi) deficiency in soils. Root architecture variables triggered by Pi availability are well documented in Arabidopsis (Arabidopsis thaliana), but the molecular mechanisms behind these adaptive responses remain to be elucidated. By the use of transcriptomic and quantitative RT-PCR analysis, we observed that an AINTEGUMENTA-like gene, named PRD for Phosphate Root Development, was rapidly repressed in roots under low Pi conditions. The physiological function of the PRD gene was analyzed through the null allele mutant prd, which displayed less development of primary and lateral roots under Pi-starvation conditions than wild-type plants. Complementation of the prd mutant with the wild-type gene led to a similar response to Pi starvation as wild-type plants, indicating the complete rescue of the mutant phenotype. These results suggest that PRD gene is involved in the regulation of root architectural responses to Pi starvation by controlling primary and lateral root elongation. This model is in agreement with the tissue-specific pattern of PRD gene expression, which was observed to occur specifically in the apex in both the primary and lateral roots. However, Pi influx, anionic profiles and root expression of genes typically induced by Pi starvation, such as high affinity Pi transporters (PHT1;1 and PHT1;4) and an acid phosphatase (AtACP5), were similar in wild type and prd plants in response to Pi starvation. These results support the hypothesis that the PRD gene is not a checkpoint for Pi-starvation responses, but acts specifically as a regulator of root architectural responses to Pi starvation. PMID- 18506480 TI - Possible involvement of MAP kinase pathways in acquired metal-tolerance induced by heat in plants. AB - Cross tolerance is a phenomenon that occurs when a plant, in resisting one form of stress, develops a tolerance to another form. Pretreatment with nonlethal heat shock has been known to protect cells from metal stress. In this study, we found that the treatment of rice roots with more than 25 muM of Cu(2+) caused cell death. However, heat shock pretreatment attenuated Cu(2+)-induced cell death. The mechanisms of the cross tolerance phenomenon between heat shock and Cu(2+) stress were investigated by pretreated rice roots with the protein synthesis inhibitor cycloheximide (CHX). CHX effectively block heat shock protection, suggesting that protection of Cu(2+)-induced cell death by heat shock was dependent on de novo protein synthesis. In addition, heat pretreatment downregulated ROS production and mitogen-activated protein kinase (MAPK) activities, both of which can be greatly elicited by Cu(2+) stress in rice roots. Moreover, the addition of purified recombinant GST-OsHSP70 fusion proteins inhibited Cu(2+)-enhanced MAPK activities in an in vitro kinase assay. Furthermore, loss of heat shock protection was observed in Arabidopsis mkk2 and mpk6 but not in mpk3 mutants under Cu(2+) stress. Taken together, these results suggest that the interaction of OsHSP70 with MAPKs may contribute to the cellular protection in rice roots from excessive Cu(2+) toxicity. PMID- 18506482 TI - Anomalous ascending aorta causing severe compression of the left bronchus in an infant with ventricular septal defect and pulmonary atresia. AB - Tracheobronchial compression by cardiovascular structures complicates the course after surgery of congenital heart disease. A 2-month-old boy with ventricular septal defect, pulmonary atresia, and velocardiofacial syndrome had severe left main bronchus obstruction due to external compression by an anomalously oriented ascending aorta. The patient's hypoxemic episodes necessitated extracorporeal membrane oxygenation. We inserted a stent in the left bronchus to open the airway, but the stent was crushed by the anomalous aorta. We later surgically rerouted the aorta and finally restored the patency of the left main bronchus. However, the patient died of fungemia, without being weaned from extracorporeal membrane oxygenation. We conclude that surgery is necessary instead of stent implantation to relieve the external compression of the left bronchus from a vessel with systemic arterial pressure. PMID- 18506481 TI - Distinct roles of the pepper pathogen-induced membrane protein gene CaPIMP1 in bacterial disease resistance and oomycete disease susceptibility. AB - Plant integral membrane proteins have essential roles in diverse internal and external physiological processes as signal receptors or ion transporters. The pepper CaPIMP1 gene encoding a putative integral membrane protein with four transmembrane domains was isolated and functionally characterized from pepper leaves infected with the avirulent strain Xanthomonas campestris pv. vesicatoria (Xcv). CaPIMP1-green fluorescence protein (GFP) fusions localized to the plasma membrane in onion cells, as observed by confocal microscopy. CaPIMP1 was expressed in an organ-specific manner in healthy pepper plants. Infection with Xcv induced differential accumulation of CaPIMP1 transcripts in pepper leaf tissues during compatible and incompatible interactions. The function of CaPIMP1 was examined by using the virus-induced gene silencing technique in pepper plants and by overexpression in Arabidopsis. CaPIMP1-silenced pepper plants were highly susceptible to Xcv infection and expressed lower levels of the defense-related gene CaSAR82A. CaPIMP1 overexpression (CaPIMP1-OX) in transgenic Arabidopsis conferred enhanced resistance to P. syringae pv. tomato infection, accompanied by enhanced AtPDF1.2 gene expression. In contrast, CaPIMP1-OX plants were highly susceptible to the biotrophic oomycete Hyaloperonospora parasitica. Taken together, we propose that CaPIMP1 plays distinct roles in both bacterial disease resistance and oomycete disease susceptibility. PMID- 18506483 TI - Mycoplasma pneumoniae: nervous system complications in childhood and review of the literature. AB - Mycoplasma pneumoniae is an important pathogen which causes nervous system disorders during or after the course of a respiratory tract infection. The exact pathogenic mechanism which causes neurological disorders still remains unknown. Although meningoencephalitis and acute disseminated encephalomyelitis are common complications, there are few cases of acute transverse myelitis and isolated abducens nerve palsy associated with M. pneumoniae infection in childhood. The association between ocular myasthenia gravis and M. pneumoniae infection has not been described before. Here, we describe five patients with different nervous system complications associated with M. pneumoniae infection and discuss the pathological features of central nervous system involvement. PMID- 18506485 TI - Placental microstructure and efficiency in cloned bovines: a design-based stereological approach. AB - Somatic nuclear transfer allows scientists to produce identical copies of individuals (clones) and offers the potential for research in various fields. However, the production of cloned bovines is inefficient, partly because of high rates of embryonic mortality associated with abnormal placental development, although the exact mechanisms are still unclear. In the present study, we have applied design-based stereological methods to quantify placental morphology in cloned and non-cloned Nelore bovines (280-297 days of gestation). We have found no differences for any of the following parameters: number and volume of placentomes, villous volume and surface area, volume and number of star volume units and placental efficiency. Therefore, placental deficiency is unlikely to be attributable to its villous component. Nevertheless, new studies should focus on other placental constituents, e.g. capillaries, trophoblast cells and feto maternal exchange surface area. PMID- 18506484 TI - Prognostic factors and treatment outcomes of patients with uterine sarcoma: analysis of 127 patients at a single institution, 1989-2007. AB - PURPOSE: Uterine sarcoma is a rare malignancy with the worst prognosis of all uterine cancers. This study evaluated the prognostic factors and treatment outcomes of patients with this disease. METHODS: A retrospective analysis was performed on 127 patients with histologically verified uterine sarcoma who were treated and followed at the Asan Medical Center (Seoul, Korea) from 1989 to 2007. RESULTS: Histological analyses revealed that 37 patients had endometrial stromal sarcoma, 44 had malignant mixed mullerian tumors and 46 had leiomyosarcoma. Surgical stages, as defined by the International Federation of Gynecology and Obstetrics (FIGO) system, were I in 82 patients, II in 6 patients, III in 18 patients and IV in 19 patients. All patients underwent surgical treatment and 72 patients received adjuvant therapy. The 10-year disease-free survival (DFS) rate was 30% and the 10-year overall survival (OS) rate was 48%, with a mean follow-up time of 38 months (ranging from 1 to 212 months). Adjuvant radiation and chemotherapy had limited impact on the outcome of early-stage disease. However, patients with advanced-stage disease who received adjuvant chemotherapy had significantly longer OS times. A multivariate analysis revealed that FIGO stage (P = 0.025), depth of myometrial invasion (P = 0.004), and complete cytoreduction (P = 0.030) were significantly associated with DFS, while menopausal status (P = 0.044), FIGO stage (P = 0.016), depth of myometrial invasion (P = 0.029), and lymph-vascular space invasion (LVSI) (P = 0.020) were significantly associated with OS. CONCLUSIONS: This study suggests that complete cytoreduction is important and adjuvant chemotherapy can help achieve favorable prognoses in patients with advanced stage disease. However, postmenopausal status, advanced FIGO stage, deep myometrial invasion, and positive LVSI were associated with poor prognosis. PMID- 18506486 TI - Effect of salmon melanin-concentrating hormone and mammalian gonadotrophin releasing hormone on somatolactin release in pituitary culture of Cichlasoma dimerus. AB - We detected a close morphological association between melanin-concentrating hormone (MCH)-immunoreactive (ir) fibers and somatolactin (SL)-ir cells in the pars intermedia of the cichlid fish Cichlasoma dimerus by double-label immunofluorescence. Male pituitaries obtained from adult C. dimerus were incubated with 0.1-10 microM salmon MCH, and the amount of SL released into the culture medium was semi-quantified by Western blot. This assay showed an increase of SL release in a dose-dependent manner (linear regression: P<0.05). A close association of GnRH-ir fibers with SL-ir cells was also detected at the pars intermedia level. Male pituitaries were also incubated with 0.1-10 microM of mammalian GnRH, and SL release was semi-quantified by Western blot, showing an increase of released SL levels in a dose-dependent manner (linear regression: P<0.05). In contrast, SL release was unaffected from female pituitaries incubated with salmon MCH; however, an increasing tendency was observed when mammalian GnRH was used. Hypothalamic close association of MCH-ir perikarya and GnRH-ir fibers was found by double-label immunofluorescence indicating a possible relationship between them. These results suggest that SL, like other pituitary hormones, is under hypothalamic control and is involved in diverse physiological processes including background adaptation and reproduction. This study has also shown that the in vitro culture of a single C. dimerus pituitary is a feasible method for studying the control of SL release and other pituitary hormones. PMID- 18506487 TI - The relationships between floral traits and specificity of pollination systems in three Scandinavian plant communities. AB - The pollination syndrome hypothesis has provided a major conceptual framework for how plants and pollinators interact. However, the assumption of specialization in pollination systems and the reliability of floral traits in predicting the main pollinators have been questioned recently. In addition, the relationship between ecological and evolutionary specialization in pollination interactions is still poorly understood. We used data of 62 plant species from three communities across southern Norway to test: (1) the relationships between floral traits and the identity of pollinators, (2) the association between floral traits (evolutionary specialization) and ecological generalization, and (3) the consistency of both relationships across communities. Floral traits significantly affected the identity of pollinators in the three communities in a way consistent with the predictions derived from the pollination syndrome concept. However, hover flies and butterflies visited flowers with different shapes in different communities, which we mainly attribute to among-community variation in pollinator assemblages. Interestingly, ecological generalization depended more on the community-context (i.e. the plant and pollinator assemblages in the communities) than on specific floral traits. While open yellow and white flowers were the most generalist in two communities, they were the most specialist in the alpine community. Our results warn against the use of single measures of ecological generalization to question the pollination syndrome concept, and highlight the importance of community comparisons to assess the pollination syndromes, and to understand the relationships between ecological and evolutionary specialization in plant pollinator interactions. PMID- 18506488 TI - Urinary aprotinin as a predictor of acute kidney injury after cardiac surgery in children receiving aprotinin therapy. AB - Proteomic analysis has revealed potential early biomarkers of acute kidney injury (AKI) in children undergoing cardiopulmonary bypass (CPB), the most prominent one with a mass-to-charge ratio of 6.4 kDa. The objective of this study was to identify this protein and test its utility as a biomarker of AKI. Trypsin digested protein bands were analyzed by tandem mass spectrometry (MS/MS) to identify the protein in urine samples. Surface-enhanced laser desorption/ionization time-of-flight analysis and a functional activity assay were performed to quantify urinary levels in a pilot study of 106 pediatric patients undergoing CPB. The protein was identified as aprotinin. Urinary aprotinin levels 2 h after initiation of CPB were predictive of AKI (for functional assay: 92% sensitivity, 96% specificity, area under the curve of 0.98). By multivariate analysis, the urinary aprotinin level 2 h after CPB was an independent predictor of AKI (beta = 0.001, P < 0.0001). The 2 h urinary aprotinin level correlated with serum creatinine, duration of AKI, and length of hospital stay. We concluded that urinary aprotinin levels 2 h after initiation of CPB predict the development of AKI and adverse clinical outcomes. PMID- 18506489 TI - Enuresis nocturna and sleep quality. AB - Enuresis nocturna is a common problem. Numerous etiologic factors have been investigated, and various theories have been proposed. The objectives of our study were to establish the differences in the sleep quality of nocturnal enuretic patients from that of healthy voluntary subjects, and the changes after treatment with desmopressin acetate (DDAVP), among primary school children. The study comprised 19 children with primary nocturnal enuresis and 32 healthy children in the control group. Subjective assessment of sleep was determined with the Pittsburgh Sleep Quality Index (PSQI) questionnaire. PSQI scores for each patient and control subject were determined before the study was started and after a month time interval. The sleep quality of the nocturnal enuretic children was poor. We found lower scores after a month's treatment with DDAVP, and significant differences in two dimensions in the patient group: 'subjective sleep quality' and 'sleep disturbances'. When we asked the patients' group what caused the sleep disturbance, they replied 'the fear or the anxiety of bedwetting during sleep'. This anxiety or fear seemed to be a factor that probably affected their sleep quality. So, active treatment (medical or behavioral) should be started as soon as the child is ready to receive it or when the enuretic child wants to be dry when asleep. PMID- 18506490 TI - Temperature, air pollution and total mortality during summers in Sydney, 1994 2004. AB - This study investigated the effect of temperature and air pollutants on total mortality in summers in Sydney, Australia. Daily data on weather variables, mortality and air pollution for the Sydney metropolitan area from 1 January 1994 to 31 December 2004 were supplied by Australian Bureau of Meteorology, Australian Bureau of Statistics, and Environment Protection Agency of New South Wales, respectively. We examined the association of total mortality with weather indicators and air pollution using generalised additive models (GAMs). A time series classification and regression tree (CART) model was developed to explore the interaction effects of temperature and air pollution that impacted on mortality. Our results show that the average increase in total daily mortality was 0.9% [95% confidence interval (CI): 0.6-1.3%] and 22% (95% CI: 6.4-40.5%) for a 1 degrees C increase in daily maximum temperature and 1 part per hundred million (pphm) increase in daily average concentration of sulphur dioxide (SO(2)), respectively. Time-series CART results show that maximum temperature and SO(2) on the current day had significant interaction effects on total mortality. There were 7.3% and 12.1% increases in daily average mortality when maximum temperature was over 32 degrees C and mean SO(2) exceeded 0.315 pphm, respectively. Daily maximum temperature was statistically significantly associated with daily deaths in Sydney during summers between 1994 and 2004. Elevated daily maximum temperature combined with high SO(2) concentrations appeared to have contributed to the increased mortality observed in Sydney during this period. PMID- 18506491 TI - Synoptic climatology of the long-distance dispersal of white pine blister rust II. Combination of surface and upper-level conditions. AB - An invasive forest pathogen, Cronartium ribicola, white pine blister rust (WPBR), is believed to have arrived in the Sacramento Mountains of south-central New Mexico about 1970. Epidemiological and genetic evidence supports the hypothesis that introduction was the result of long-distance dispersal (LDD) by atmospheric transport from California. This study applies a method to identify the atmospheric conditions favorable for rust transport and infection. An upper level synoptic classification (ULSC) identifies patterns of upper-level flow favorable for the transport of rust spores from a source to a target. Transport data are coupled with data for surface conditions favorable for infection at a designated target. A resulting calendar lists likelihood classes for establishment by four times-daily observations during a dispersal season from April through July in the years 1965 to 1974. The single most-favorable period for transport and infection at the New Mexico site was identified as 1-15 June 1969. Five additional sites in the western United States with susceptible white pine populations and known infestation status were then evaluated to verify the model. Only the infested sites exhibit an establishment likelihood of "high" or "very high." This suggests that the methodology correctly identifies locations with elevated establishment likelihood. Finally, likelihoods at nine additional points in the southwestern United States are determined and used to map regional patterns of transport, infection and establishment. The ULSC combined with appropriate surface meteorological data could be used to further investigate transport and infection, identify other areas at risk, assess the potential for gene flow of WPBR and evaluate long-distance dispersal of other pathogens. PMID- 18506492 TI - Decline in the workload associated with the serodiagnosis of syphilis in a general hospital: 1994-2004. AB - Syphilis re-emergence is a cause of concern. Our objective was to quantify the laboratory workload, incident cases and patient follow-up that syphilis has generated for 11 years in a large teaching hospital. An ecologic study including all samples submitted for syphilis serodiagnosis at our hospital from January 1994 to December 2004 was undertaken. Our laboratory processed 58,832 samples for syphilis serodiagnosis. From 1994 to 2004, the number of samples submitted for syphilis testing dropped by 11% (95% confidence interval [CI] 10-12, p<0.001). Syphilis was diagnosed in 443 patients. The incidences were 11, 3 and 8 per 100,000 inhabitants/year in 1994, 2000 and 2004, respectively. Only 42% (185) of patients had repeat tests and in 79% (146) of cases, the 1-year follow-up data were missing. The median follow-up was 9 months (interquartile range [IQR] 3-26). We detected a reduction in the effort to detect syphilis, despite an increase in its incidence and the low cost of syphilis screening. Efforts should be intensified to improve physician compliance with syphilis screening and follow-up guidelines. PMID- 18506493 TI - Commonly missed subtle skeletal injuries in children: a pictorial review. AB - Children are distinctive as compared to adults when it comes to musculoskeletal injuries. This is due to the relative elasticity of bones and the presence of epiphyseal plates. There are many subtle injuries which will be missed if the radiologist is not aware of them and is not actively searching for them. The common elusive injuries include: (1) plastic bending fractures, (2) sternoclavicular dislocation, (3) epiphyseal-metaphyseal injuries in older child, (4) buckle fractures, and (5) Toddler fracture types I and II. Detection of these injuries needs an accurate history, a good physical examination, and, in particular, a thorough search by the radiologist. In many cases, it is the radiologist who suggests likelihood of the injury and guides management. In this respect, the use of comparative views and, in some cases, additional imaging is warranted. Here, we review the elusive musculoskeletal injuries in children in pictorial form. PMID- 18506494 TI - Surgery in adult onset tethered cord syndrome (ATCS): review of literature on occasion of an exceptional case. AB - Tethered cord syndrome, usually discovered in childhood, is a developmental abnormality impairing the longitudinal movement of the spinal cord that can be combined with various forms of spinal dysraphism. Adult onset tethered cord syndrome (ATCS) seems not as rare as once thought, however, low susceptibility in adulthood commonly leads to a delay in diagnosis and therapy. We conducted a meticulous literature research to evaluate the clinical presentation, associated malformations, prognostic factors, as well as the benefits and risks of surgical treatment in ATCS patients. The age of patients at onset of symptoms ranged from 18 to 76 years with a mean of 36.5 years, including 184 males and 202 females. In contrast to the pediatric clientele, pain is the predominant symptom in adults, and elicitating mechanisms like trauma, excessive physical training, or degenerative spinal canal stenosis are reported more often. Surgical untethering aims the restoration of craniocaudal mobility of the spinal cord in order to prevent the further progression of symptoms, to restore neurological function, and to improve pain. In our evaluation of literature, pain was the most responsive symptom after surgical untethering (307 of 368 patients). Sensory and motor symptoms also seem to benefit from the surgery, especially if less than 6 months standing and mild. Improvement could be achieved in 43% (144 of 335 patients) for sensory deficits and 58.6% (191 of 326 patients) for motor deficits. Sphincter troubles are less responsive; they show an improvement in 45.6% (141 of 309 patients). Factors reported to be associated with the postoperative deterioration and/or bad outcome are the split cord malformation, lipomyelomeningocele, previous surgery, rapid motor function worsening experienced shortly before the operation, and long delay in diagnosis. The rate of secondary decline and retethering could not be established in our literature research. We report on an additional case of ATCS with the late onset of symptoms at the age of 49; she underwent surgical untethering with neuronavigational guidance. Performing neuronavigational guidance on the basis of multimodal images (computed tomography and magnetic resonance imaging fused with the intraoperative biplanar X-ray) in our patient showed substantial benefit in the surgical orientation within a complex skeletal and neuronal anomaly. PMID- 18506495 TI - Staphylococcus aureus enterotoxin C2 mutants: biological activity assay in vitro. AB - Staphylococcal enterotoxin C2 (SEC2) is one member of bacterial superantigens produced by Staphylococcus aureus. It can be attributed to its superantigenic activity to cross-link major histocompatibility complex class II molecules with T cell receptors and activate a large number of resting T cells resulting in release of massive cytokines, which will produce significant tumor inhibition in vivo and in vitro. However, it could be not broadly applied to cure malignant tumors in clinic because of emetic activity of SEC2. The aim of this study was to inactivate emetic activity of SEC2 through site-directed mutagenesis. Cys93, Cys110 and His118 were selected as substitutional sites based on the functional sites responsible for emesis. The mutated proteins were used to determine Peripheral blood mononuclear cell proliferation activity and anti-tumor activity in vitro. Results showed that these mutated proteins efficiently stimulated T cell and exhibited the same tumor-inhibition effect as SEC2. It is possible to inactivate emetic activity of SEC2 through site-directed mutagenesis and provide satisfying agents for tumor treatment in clinic. PMID- 18506496 TI - Polyhexamethyl biguanide can eliminate contaminant yeasts from fuel-ethanol fermentation process. AB - Industrial ethanol fermentation is a non-sterile process and contaminant microorganisms can lead to a decrease in industrial productivity and significant economic loss. Nowadays, some distilleries in Northeastern Brazil deal with bacterial contamination by decreasing must pH and adding bactericides. Alternatively, contamination can be challenged by adding a pure batch of Saccharomyces cerevisiae-a time-consuming and costly process. A better strategy might involve the development of a fungicide that kills contaminant yeasts while preserving S. cerevisiae cells. Here, we show that polyhexamethyl biguanide (PHMB) inhibits and kills the most important contaminant yeasts detected in the distilleries of Northeastern Brazil without affecting the cell viability and fermentation capacity of S. cerevisiae. Moreover, some physiological data suggest that PHMB acts through interaction with the yeast membrane. These results support the development of a new strategy for controlling contaminant yeast population whilst keeping industrial yields high. PMID- 18506497 TI - Occurrence and temporal variations of the xenoestrogens bisphenol A, 4-tert octylphenol, and tech. 4-nonylphenol in two German wastewater treatment plants. AB - GOAL, SCOPE, AND BACKGROUND: The xenoestrogens bisphenol A, 4-tert-octylphenol, and the technical isomer mixture of 4-nonylphenol (tech. 4-nonylphenol) belong to the group of chemicals which are called endocrine disrupters due to their property of causing hormonal dysfunctions in the endocrine system of organisms at very low concentrations. Bisphenol A, 4-tert-octylphenol, and the tech. 4 nonylphenol (mixture of isomers) were determined in water samples collected from the influent and effluent of two German wastewater treatment plants (WWTP) during a long-time sampling period from February 2003 till August 2005 to assess their occurrence and temporal variations in WWTPs. METHODS: The compounds were extracted and concentrated from water by solid-phase extraction (SPE) using Bond Elut PPL cartridges and quantified by use of gas chromatography-mass spectrometry (GC-MS). RESULTS: The influent concentrations were as follows: Bisphenol A < limit of detection of the method (< ldm)--12,205 ng L(-1), tech. 4-nonylphenol < ldm--10,186 ng L(-1), and 4-tert-octylphenol 39-1,495 ng L(-1). The measured effluent concentrations were lower with values in the range of < ldm--7,625 ng L( 1) for bisphenol A, < ldm--14,444 ng L(-1) for tech. 4-nonylphenol, and < ldm- 392 ng L(-1) for 4-tert-octylphenol. All target compounds were largely eliminated during the wastewater treatment process. The elimination efficiency varied between 73% and 93%. DISCUSSION: All analytes show highly fluctuating influent concentrations with very high peak concentrations at particular sampling times. The variation of effluent concentrations is by far lower than the variation of influent concentrations. For tech. 4-nonylphenol, a significant temporal concentration variation has been detected with very high concentrations up to the microgram-per-liter level in the time from February 2003 till July 2003 and clearly decreasing concentrations in the time from June 2004 till August 2005. This corresponds well with the implementation of Directive 2003/53/EC (nonylphenol and nonylphenol ethoxylates in the European Union "may not be placed on the marked or used as a substance or constituent of preparations in concentrations equal or higher than 0.1% by mass") from January 2005 on. Bisphenol A is present in the effluent samples in a wide range of concentrations from below the detection limit to high concentrations up to the microgram-per liter level. For 4-tert-octylphenol, no particular trend of concentration development has been observed. CONCLUSIONS: Combined SPE and GC-MS proved to be an efficient method to identify and quantify polar organic compounds in environmental samples. With respect to the concentrations measured in the present study, bisphenol A sometimes is the prominent compound in influent samples. Neither bisphenol A nor 4-tert-octylphenol or tech. 4-nonylphenol show seasonal variations. However, there was a significant general trend of decreasing concentrations of tech. 4-nonylphenol in influent and effluent samples from both WWTPs which probably reflects the implementing Directive 2003/53/EC. RECOMMENDATIONS AND PERSPECTIVES: Further research is needed to investigate whether the observed decrease of tech. 4-nonylphenol concentrations in German WWTPs since June 2004 will continue further on. The reason for the high effluent concentrations of bisphenol A in only a few samples has to be clarified in further research. The results from this study provide insight into the concentration development of the xenoestrogens bisphenol A, tech. 4-nonylphenol, and 4-tert-octylphenol in WWTPs in the time span between 2003 and 2005. PMID- 18506498 TI - The mitochondrial genome of a deep-sea bamboo coral (Cnidaria, Anthozoa, Octocorallia, Isididae): genome structure and putative origins of replication are not conserved among octocorals. AB - Octocoral mitochondrial (mt) DNA is subject to an exceptionally low rate of substitution, and it has been suggested that mt genome content and structure are conserved across the subclass, an observation that has been supported for most octocorallian families by phylogenetic analyses using PCR products spanning gene boundaries. However, failure to recover amplification products spanning the nad4L msh1 gene junction in species from the family Isididae (bamboo corals) prompted us to sequence the complete mt genome of a deep-sea bamboo coral (undescribed species). Compared to the "typical" octocoral mt genome, which has 12 genes transcribed on one strand and 5 genes on the opposite (cox2, atp8, atp6, cox3, trnM), in the bamboo coral genome a contiguous string of 5 genes (msh1, rnl, nad2, nad5, nad4) has undergone an inversion, likely in a single event. Analyses of strand-specific compositional asymmetry suggest that (i) the light-strand origin of replication was also inverted and is adjacent to nad4, and (ii) the orientation of the heavy-strand origin of replication (OriH) has reversed relative to that of previously known octocoral mt genomes. Comparative analyses suggest that intramitochondrial recombination and errors in replication at OriH may be responsible for changes in gene order in octocorals and hexacorals, respectively. Using primers flanking the regions at either end of the inverted set of five genes, we examined closely related taxa and determined that the novel gene order is restricted to the deep-sea subfamily Keratoisidinae; however, we found no evidence for strand-specific mutational biases that may influence phylogenetic analyses that include this subfamily of bamboo corals. PMID- 18506499 TI - Effects of ethinylestradiol on medaka (Oryzias latipes) as measured by sperm motility and fertilization success. AB - We investigated the effects of 30-480 ng/L 17alpha-ethinylestradiol (EE(2)) on the sperm motility and fertility of Japanese medaka (Oryzias latipes). Sperm motility was examined by computer-assisted image analysis. In male medaka, the velocity of sperm was found to have increased after 3 weeks of exposure at 60-480 ng/L. This result suggests that higher sperm velocities depleted sperm energy reserves more rapidly and shortened the time for which sperm were viable to fertilize eggs. In a separate experiment that studied whether EE(2) exposure of males affects the fertilization rate or hatchability, sexually mature male medaka were exposed for 3 weeks and subsequently evaluated for their reproductive ability after pairing with unexposed females for 7 days. Exposure of males to EE(2) exerted a potent inhibitory effect on a reproduction parameter (fertilization rate x hatchability), and the highest inhibition was observed at 60 ng/L. The results offer toxicological data for the assessment of EE(2 )exposure in medaka and suggest that short-term exposure to EE(2) might reduce sperm function and fertility in adult male medaka. PMID- 18506500 TI - DNA-protein cross-links in erythrocytes of freshwater fish exposed to hexavalent chromium or divalent nickel. AB - DNA-protein cross-links (DPXs) in fish erythrocytes represent a potential biomarker for exposure to metal cations, such as hexavalent chromium (Cr[VI]) and divalent nickel (Ni[II]). Species-specific sensitivities to DPX formation were studied by coexposure of juvenile specimens of rainbow trout, hybrid bluegill, and channel catfish to waterborne metals, such as Cr(VI) and Ni(II). In a species comparison, 4 days of exposure to 2 ppm Cr(VI) induced highest DPXs in bluegill erythrocytes, followed by trout and catfish, at 186%, 97%, and 48% above controls, respectively. A similar pattern of species sensitivity was observed following co-exposure of the fish to 15 ppm Ni(II) for 4 days, with 237%, 124%, and 82% increased DPXs above control bluegill, trout, and catfish, respectively. Biological stability of Cr(VI)-induced DPXs was demonstrated in Cr(VI)-exposed bluegill, as DPX levels remained elevated for up to 20 days after discontinuation of exposure. Similar results were found following exposure of catfish to Ni(II), with detectable DPXs found 10 days after acute exposure. In both bluegill and catfish, a continued increase in DPX formation in erythrocytes was seen for 5-10 days after Cr(VI) was removed from tank water, suggesting that residual Cr(VI) may be involved in DPX formation following acute exposure of fish. PMID- 18506501 TI - Assessment of myocardial function in pediatric patients with operated tetralogy of Fallot: preliminary results with 2D strain echocardiography. AB - The global myocardial function in patients after repair of tetralogy of Fallot (TOF) can be assessed by cardiovascular magnetic resonance (CMR) and measurement of B-type natriuretic peptides. Two-dimensional echocardiography-derived strain and strain rate (2D strain) facilitate the assessment of regional myocardial function. We evaluated myocardial function in 16 children with residual severe pulmonary valve regurgitation and right ventricular (RV) volume overload after TOF repair before, 1 month after, and 6 months after pulmonary valve replacement (PVR). In 2D strain echocardiography preoperatively, the longitudinal systolic RV strain was reduced (p < 0.05). One month after PVR, longitudinal systolic RV strain decreased further (p < 0.05), while systolic and early diastolic radial left ventricular strain and strain rate increased (each p < 0.05), followed by a return toward preoperative values after 6 months. Six months after PVR, preoperatively elevated RV end-diastolic volume (p < 0.01) assessed by CMR and N terminal pro-B-type natriuretic peptide (p < 0.05) decreased. In conclusion, the impairment of the regional myocardial after TOF repair and transient changes after PVR can be subtly analyzed by 2D strain echocardiography in addition to the established assessment of myocardial function with CMR and measurement of B-type natriuretic peptides. PMID- 18506502 TI - Role of plants in the vegetative and reproductive growth of saprobic basidiomycetous ground fungi. AB - Non-symbiotic microorganisms engineered or expensively selected to degrade xenobiotic hydrocarbons or modify heavy-metal uptake of plants in soil remediations die back after their introduction into the target soils. Mycelia of saprobic basidiomycetes were therefore inoculated into soil samples of 1 l in glass vessels to record mycelial growth and reproduction in the immediate rhizosphere of up to 11 herbaceous plant species, or to study their responses to the separate volatiles from whole plant swards or their root balls whose emanations had been collected in 1.5-l plastic bags fixed to the glass vessels. Excess CO2 was controlled with NaOH solution. Volatiles from root balls of parsley and pea but not wheat, from unplanted soils, from the fungus-permeated, unplanted substrate soil itself, and from the rooting soil of whole wheat sward increased mycelial densities in Clitocybe sp. more than in Agaricus macrocarpus and indicated thus a higher nutrient state of the mycelia. Organic volatiles proved therefore to be a significant carbon source for certain basidiomycetes in poor natural soils. The contemporary decline in the number of basidiocarp initials to 0 to 36% in both fungi relative to the unplanted and aerated controls was caused by volatiles from rooted and unplanted soil and pointed thus to their ecological role as antibiotics, fumigants, toxins, and hormonal compounds. Aqueous extracts from root balls of wheat stimulated mycelial density and fruiting in A. macrocarpus contemporarily because of their contents in soil derived macronutrients. They suppressed once more fruiting in the more sensitive Clitocybe sp. by active agents in the aqueous phase. Within plant rhizospheres, densities of Clitocybe sp. mycelia were stimulated in the presence of alfalfa, carrot, red clover, ryegrass, and spinach, whereas those of A. macrocarpus were halved by 7 of 10 plant species including alfalfa, red clover, ryegrass, and spinach. Mycelia of A. macrocarpus may thereby have responded to differences in concentration and composition of volatile compounds. The contemporary repression of fruiting in both fungi and in nearly all treatments was not due to plant competition for macronutrients. Mycelia of basidiomycetes over-compensated for losses in macronutrients to the plant by decomposing soil matrix constituents. It is concluded that organic volatiles emitted by several plant organs and natural soils improved the nutritional state of A. macrocarpus and Clitocybe sp. but not of Agaricus bisporus mycelia and could therefore help establish certain ground fungi in the field. The contemporary and general suppression of fruiting by constituents of the gaseous (and liquid) phase in all fungi examined suggests interference with basic physiological processes and recommends an urgent re examination of the degradative ability of basidiomycetes in the presence of volatiles. PMID- 18506503 TI - Peritumoral steatosis associated with insulinomas: appearance at imaging. AB - We report three patients with insulinoma tumors and distinct peritumoral steatosis, well demonstrated with several imaging modalities. This unique appearance can aid in pre-operative diagnosis and guide biopsy. PMID- 18506504 TI - Percutaneous paracoccygeal catheter drainage of deep pelvic abscesses using a combination of sonographic and fluoroscopic guidance. AB - BACKGROUND: The purpose of this study is to demonstrate the feasibility, safety, and success of percutaneous paracoccygeal catheter drainage of deep pelvic abscesses using a combination of sonography and fluoroscopic guidance. METHODS: From April 2005 to December 2006, under sonography and fluoroscopic guidance, sixteen patients with deep pelvic abscess underwent percutaneous paracoccygeal drainage. The causes of pelvic abscesses were post-operative complications. The patient was in prone position; the puncture site was first evaluated with sonography. After local anesthesia, a thin needle (22 G) was positioned along the lateral margin of the coccyx to avoid an eventual injury of sciatic nerve or vessels into the deep pelvic abscess under sonography guidance. Small dose of contrast was injected under fluoroscopy to verify the needle. With Seldinger technique catheters were placed for drainage. Catheter drainage was maintained for 9-40 days. Patients underwent clinical follow-up and subsequent imaging as necessary. RESULTS: Neither significant complications nor technical failures were observed. A complete resolution of the abscess was achieved in 14 patients, and significant resolution of abscess was also observed in the other 2 patients. CONCLUSIONS: Sonography and fluoroscopic guided percutaneous paracoccygeal drainage is safe, feasible and efficient approach in the treatment of deep pelvic abscess. PMID- 18506509 TI - A new application for increasing breast projection in free-nipple-graft reduction mammaplasty. AB - BACKGROUND: Free-nipple-graft reduction mammaplasty is the procedure of choice in patients with massive breast hypertrophy. Breasts that have undergone free-nipple graft reduction mammaplasty usually cannot maintain projection. Many modifications of free-nipple-graft reduction mammaplasty have been described to treatment this problem. We describe our modification of the free-nipple-graft reduction mammaplasty. METHODS: The technique includes two key points: The first is the fold of the midportions of the breast. The second is suturing the midportion of the breast to the fascia of the pectoral muscle. The fold increases breast projection and the suturing helps to obtain long-lasting breast projection. RESULTS: A total of 16 patients underwent free-nipple-graft reduction with the modified technique between 2003 and 2008. Mean follow-up was 21 months (range = 3-60 months). An average of 2016 g of tissue per breast was excised (range = 1250-2700 g per breast). An average of 4031 g of tissue per patient was excised. All patients had long-lasting, pronounced breast mound projection. Satisfactory breast projection was maintained. No "bottoming out" was seen. The level of satisfaction felt by all patients was very high. CONCLUSION: Long-term projection can be maintained by suturing the half-trianges to each other and to the fascia of the pectoral muscle. One more useful alternative technique for increasing projection in free-nipple-graft reduction has been added with the technique presented. PMID- 18506510 TI - The "jaguar's eye" as a new beauty trend? Age-related effects in judging the attractiveness of the oblique eye axis. AB - BACKGROUND: The eye area plays an important role in the assessment of a person. Although plastic surgery in this area is quite common, only a few studies have evaluated the features that create an attractive eye. This study aimed to determine whether a preference exists for a certain eye axis. METHODS: The stimulus material comprised portrait images of seven women. Two versions of each face were generated that differed only in the position of the eye axis (normal vs rotated). The eye axis was rotated by raising the lateral canthus of the eye about 5 degrees . A total of 250 experimental subjects ranging in age from 15 to 84 years stated their preferential position of the eye axis (horizontal vs oblique). RESULTS: Clear evidence showed that age has an impact on the type of eye axis preferred. Young subjects (age, B 19 years) regarded the oblique type of eye axis as more attractive, whereas older subjects (age, C50 years) preferred the eyes more horizontal (p ? 0.001), independently of the interviewees' gender. CONCLUSION: Because young people judge eye attractiveness differently than older people and because beauty trends are normally determined by young people, a future ideal of beauty could be characterized by eyes with a gently oblique eye axis. PMID- 18506511 TI - Facial rejuvenation with SMASectomy and FAME using vertical vectors. AB - The quest for better results in the midface after a face lift has led to the repositioning of a structure called the malar fat pad. Finger-assisted malar elevation (FAME) consists of detaching the malar fat pad from the underlying SMAS, which allows for the elevation of this structure. Two hundred five patients (189 females and 16 males) from January 2002 to August 2007 underwent a facial rejuvenation procedure comprising short-scar rhytidoplasty, SMASectomy, and FAME, with or without a simultaneous endobrow, blepharoplasty, and lipofilling. The midface fixation technique consisted of a stitch from the malar fat pad and SMAS flap to the periosteum at the zygomatic arch which was performed in every case. Elevation of the midface and improvement of the nasolabial fold and the mandible contour were obtained in all cases. Facial aging should be evaluated as a global process instead of a segmented one. Aging occurs in every structure of the face in different ways, depending on the vector of descent, thereby treatment must be individualized. We have observed improvement of the midface when using the FAME procedure in a rhytidoplasty with SMASectomy with deep fixation. PMID- 18506512 TI - Effects of pulsed electromagnetic fields on postoperative pain: a double-blind randomized pilot study in breast augmentation patients. AB - BACKGROUND: Postoperative pain may be experienced after breast augmentation surgery despite advances in surgical techniques which minimize trauma. The use of pharmacologic analgesics and narcotics may have undesirable side effects that can add to patient morbidity. This study reports the use of a portable and disposable noninvasive pulsed electromagnetic field (PEMF) device in a double-blind, randomized, placebo-controlled pilot study. This study was undertaken to determine if PEMF could provide pain control after breast augmentation. METHODS: Forty-two healthy females undergoing breast augmentation for aesthetic reasons entered the study. They were separated into three cohorts, one group (n = 14) received bilateral PEMF treatment, the second group (n = 14) received bilateral sham devices, and in the third group (n = 14) one of the breasts had an active device and the other a sham device. A total of 80 breasts were available for final analysis. Postoperative pain data were obtained using a visual analog scale (VAS) and pain recordings were obtained twice daily through postoperative day (POD) 7. Postoperative analgesic medication use was also followed. RESULTS: VAS data showed that pain had decreased in the active cohort by nearly a factor of three times that for the sham cohort by POD 3 (p < 0.001), and persisted at this level to POD 7. Patient use of postoperative pain medication correspondingly also decreased nearly three times faster in the active versus the sham cohorts by POD 3 (p < 0.001). CONCLUSION: Pulsed electromagnetic field therapy, adjunctive to standard of care, can provide pain control with a noninvasive modality and reduce morbidity due to pain medication after breast augmentation surgery. PMID- 18506513 TI - Estimating the economic value of national parks with count data models using on site, secondary data: the case of the great sand dunes national park and preserve. AB - We estimate an individual travel cost model for Great Sand Dunes National Park and Preserve (GSD) in Colorado using on-site, secondary data. The purpose of the on-site survey was to help the National Park Service better understand the visitors of GSD; it was not intended for a travel cost model. Variables such as travel cost and income were estimated based on respondents' Zip Codes. Following approaches found in the literature, a negative binomial model corrected for truncation and endogenous stratification fit the data the best. We estimate a recreational benefit of U.S. $89/visitor/year or U.S. $54/visitor/24-h recreational day (in 2002 U.S. $). Based on the approach presented here, there are other data sets for national parks, preserves, and battlefields where travel cost models could be estimated and used to support National Park Service management decisions. PMID- 18506514 TI - Life cycle considerations for improving sustainability assessments in seafood awareness campaigns. AB - It is widely accepted that improving the sustainability of seafood production requires efforts to reverse declines in global fisheries due to overfishing and to reduce the impacts to host ecosystems from fishing and aquaculture production technologies. Reflective of on-going dialogue amongst participants in an international research project applying Life Cycle Assessment to better understand and manage global salmon production systems, we argue here that such efforts must also address the wider range of biophysical, ecological, and socioeconomic impacts stemming from the material and energetic throughput associated with these industries. This is of particular relevance given the interconnectivity of global environmental change, ocean health, and the viability of seafood production in both fisheries and aquaculture. Although the growing popularity of numerous ecolabeling, certification, and consumer education programs may be making headway in influencing Western consumer perceptions of the relative sustainability of alternative seafood products, we also posit that the efficacy of these initiatives in furthering sustainability objectives is compromised by the use of incomplete criteria. An emerging body of Life Cycle Assessment research of fisheries and aquaculture provides valuable insights into the biophysical dimensions of environmental performance in alternative seafood production and consumption systems, and should be used to inform a more holistic approach to labeling, certifying, and educating for sustainability in seafood production. More research, however, must be undertaken to develop novel techniques for incorporating other critical dimensions, in particular, socioeconomic considerations, into our sustainability decision-making. PMID- 18506515 TI - Stories and maps, images and archives: multimethod approach to the political ecology of native property rights and natural resource management in Sabah, Malaysia. AB - The study of human-environmental relations is complex and by nature draws on theories and practices from multiple disciplines. There is no single research strategy or universal set of methods to which researchers must adhere. Particularly for scholars interested in a political ecology approach to understanding human-environmental relationships, very little has been written examining the details of "how to" design a project, develop appropriate methods, produce data, and, finally, integrate multiple forms of data into an analysis. A great deal of attention has been paid, appropriately, to the theoretical foundations of political ecology, and numerous scholarly articles and books have been published recently. But beyond Andrew Vayda's "progressive contextualization" and Piers Blaikie and Harold Brookfield's "chains of explanation," remarkably little is written that provides a research model to follow, modify, and expand. Perhaps one of the reasons for this gap in scholarship is that, as expected in interdisciplinary research, researchers use a variety of methods that are suitable (and perhaps unique) to the questions they are asking. To start a conversation on the methods available for researchers interested in adopting a political ecology perspective to human-environmental interactions, I use my own research project as a case study. This research is by no means flawless or inclusive of all possible methods, but by using the details of this particular research process as a case study I hope to provide insights into field research that will be valuable for future scholarship. PMID- 18506516 TI - Livelihood change and livelihood sustainability in the uplands of Lembang subwatershed, West Sumatra, Indonesia, in a changing natural resource management context. AB - This paper analyzes livelihood change and livelihood sustainability of households in the upland part of the Lembang subwatershed, West Sumatra, in response to changes in the natural resource management context during the last decade. Using the sustainable livelihood framework (SLF), we measured livelihood changes at two separate points in time, 1996 and 2006, and assessed their environmental, economic, social, and institutional sustainability. We found that people with a low income had less access to capital assets than people from middle- and high income groups. Our analysis revealed, however, that access to capital assets increased over time, and that poor households experienced economic improvement, indicating an overall increase in economic sustainability. Environmental sustainability, however, is threatened by intensive agricultural practices such as high agrochemical input and intensive soil tillage on steep slopes, leading to pollution and soil erosion. Social sustainability is also a matter of concern: while social exclusion has been reduced, income inequity has increased. Institutional sustainability is likely to remain uncertain, as local institutions for natural resource management are still weak, despite the fact that decentralization has been implemented during the last 8 years. External facilitation is needed to improve the livelihood of upland people while, at the same time, enhancing the sustainability of watershed management. Strengthening local institutions, conserving natural resources, and promoting environmentally sound agricultural practices are the three most important policies to be promoted within the watershed. PMID- 18506517 TI - A framework for fully integrating environmental assessment. AB - A new framework for environmental assessment is needed because no existing framework explicitly includes all types of environmental assessments. We propose a framework that focuses on resolving environmental problems by integrating different types of assessments. Four general types of assessments are included: (1) condition assessments to detect chemical, physical, and biological impairments; (2) causal pathway assessments to determine causes and identify their sources; (3) predictive assessments to estimate environmental, economic, and societal risks, and benefits associated with different possible management actions; and (4) outcome assessments to evaluate the results of the decisions of an integrative assessment. The four types of assessments can be neatly arrayed in a two-by-two matrix based on the direction of analysis of causal relationships (rows) and whether the assessment identifies problems or solves them (columns). We suggest that all assessments have a common structure of planning, analysis, and synthesis, thus simplifying terminology and facilitating communication between types of assessments and environmental programs. The linkage between assessments is based on intermediate decisions that initiate another assessment or a final decision signaling the resolution of the problem. The framework is applied to three cases: management of a biologically impaired river, remediation of a contaminated site, and reregistration of a pesticide. We believe that this framework clarifies the relationships among the various types of assessment processes and their links to specific decisions. PMID- 18506518 TI - Skewed riskscapes and environmental injustice: a case study of metropolitan St. Louis. AB - This article presents a case study of Toxics Release Inventory (TRI) air pollution exposure risks across metropolitan St. Louis. The first section critically reviews environmental justice research and related barriers to environmental risk management. Second, the paper offers a conventional analysis of the spatial patterns of TRI facilities and their surrounding census block group demographics for metropolitan St. Louis. Third, the article describes the use of an exposure risk characterization for 319 manufacturers and their air releases of more than 126 toxic pollutants. This information could lead to more practical resolutions of urban environmental injustices. The analysis of TRIs across metropolitan St. Louis shows that minority and low-income residents were disproportionately closer to industrial pollution sources at nonrandom significance levels. Spatial concentrations of minority residents averaged nearly 40% within one kilometer of St. Louis TRI sites compared to 25% elsewhere. However, one-fifth of the region's air pollution exposure risk over a decade was spatially concentrated among only six facilities on the southwestern border of East St. Louis. This disproportionate concentration of some of the greatest pollution risk would never be considered in most conventional environmental justice analyses. Not all pollution exposure risk is average, and the worst risks deserve more attention from environmental managers assessing and mitigating environmental injustices. PMID- 18506519 TI - Radiofrequency ablation of colorectal liver metastases: small size favorably predicts technique effectiveness and survival. AB - The objective of this study was to analyze long-term results of radiofrequency thermal ablation (RFA) for colorectal metastases (MTS), in order to evaluate predictors for adverse events, technique effectiveness, and survival. One hundred ninety-nine nonresectable MTS (0.5-8 cm; mean, 2.9 cm) in 122 patients underwent a total of 166 RFA sessions, percutaneously or during surgery. The technique was "simple" or "combined" with vascular occlusion. The mean follow-up time was 24.2 months. Complications, technique effectiveness, and survival rates were statistically analyzed. Adverse events occurred in 8.1% of lesions (major complication rate: 1.1%), 7.1% with simple and 16.7% with combined technique (p = 0.15). Early complete response was obtained in 151 lesions (81.2%), but 49 lesions (26.3%) recurred locally after a mean of 10.4 months. Sustained complete ablation was achieved in 66.7% of lesions < or = 3 cm versus 33.3% of lesions > 3 cm (p < 0.0001). Survival rates at 1, 3, and 5 years were 91%, 54%, and 33%, respectively, from the diagnosis of MTS and 79%, 38%, and 22%, respectively, from RFA. Mean survival time from RFA was 31.5 months, 36.2 in patients with main MTS < or = 3 cm and 23.2 in those with at least one lesion > 3 cm (p = 0.006). We conclude that "simple" RFA is safe and successful for MTS < or = 3 cm, contributing to prolong survival when patients can be completely treated. PMID- 18506520 TI - Intra-arterial calcium gluconate treatment after hydrofluoric acid burn of the hand. AB - Hydrofluoric acid (HF) is a colorless corrosive acid used in different industrial branches. Exposure to HF typically results from spills, and most often the hand or fingers are involved. Tissue damage through cutaneous HF exposure occurs through corrosive burns due to the free hydrogen ions and through skin penetration of the fluoride ions, causing a depletion of calcium in the deep tissue layers, ultimately leading to cell death and tissue necrosis. Treatment of HF burns consists of thoroughly flushing the exposed area with water and applying calcium gluconate gel to the skin. If topical treatment does not suffice, subcutaneous injections, as well as intravascular--both intravenous and intra arterial--calcium gluconate therapy, have been advocated. We report for the first time a case of HF burn of the hand and digits associated with vasospasm. Pain and vasospasm were successfully treated by repeated intra-arterial calcium gluconate injection. We conclude that intra-arterial calcium gluconate injection is a successful and well-tolerated therapy for HF burn associated with Raynaud's syndrome. Intra-arterial injection allows for well-controlled delivery of therapy as well as assessment of the vascular status. PMID- 18506521 TI - Treatment of an unusual complication of endovenous laser therapy: multiple small arteriovenous fistulas causing complete recanalization. AB - A 67-year-old woman was admitted to our institution with pain, night cramping, and visible varicose veins on her left leg. Doppler ultrasonography revealed continuous reflux in the great saphenous vein when the patient did the Valsalva maneuver. Endovenous laser therapy was applied to the great saphenous vein. Doppler ultrasonography 7 days later showed recanalization of, and arterialized flow in, the great saphenous vein. There also were small arterial vessels adjunct to the recanalized side. A left femoral angiography via a right femoral approach showed multiple small arteriovenous fistulas between superficial femoral artery muscle branches and the great saphenous vein. A second endovenous laser treatment was done at 80 J/cm, but the recanalization persisted. We offered to treat this endovascularly, but the patient preferred a surgical option. To the best of our knowledge, this is the first report of the demonstration of such a complication with endovenous laser therapy. PMID- 18506522 TI - Laceration of the common femoral artery following deployment of the starclose vascular closure system. AB - StarClose is a novel arterial closure device which achieves hemostasis, following arteriotomy, via a nitinol clip deployed on the outer arterial wall. Since its introduction to the market, several studies have shown StarClose to be both safe and effective, with few major complications encountered. We report a case of common femoral artery laceration following deployment of the StarClose vascular closure system. We conclude that the injury occurred secondary to intravascular misplacement of the nitinol clip. PMID- 18506523 TI - Ferrous, but not ferric, iron maintains homeostasis in Histoplasma capsulatum triacylglycerides. AB - Iron is an indispensable micronutrient for virtually all microorganisms, where it acts as a cofactor of many enzymes involved in regulation of multiple cellular and physiological functions. This metal is also considered an important determinant contributing to the pathogenesis of fungal infectious diseases, and therefore the identification of iron-regulated metabolic processes occurring within the invading fungal cell can help the development of new antifungal therapeutic strategies. In this study, we examined relationships between iron availability and neutral storage lipids in Histoplasma capsulatum, a dimorphic fungus responsible for the most common respiratory and systemic mycosis in humans. Yeast cells were grown in a defined minimal medium supplemented with or without iron. Lipids were extracted from cells at the log and late stationary growth phases, then separated by thin-layer chromatography, and fatty acids were analyzed by gas chromatography. A culture age-related decrease in the unsaturated fatty acid content was observed in all four neutral lipid classes examined. Iron related alterations could be seen in relation to triacylglycerol and free fatty acid pools, whereas no iron-dependent effects were detected in diacylglycerol and steryl ester fractions. Regarding triacylglycerols, the presence of iron positively affected the content of unsaturated fatty acids, and this stabilizing action of iron was notably increased when ferrous ions were added. Subsequent iron uptake studies showed a definite preference of H. capsulatum to acquire iron in its reduced, more soluble, ferrous form, and therefore, the availability of iron may be the underlying reason for the observed iron-maintained homeostasis in H. capsulatum triacylglycerols. PMID- 18506525 TI - NOTES: of caution. PMID- 18506526 TI - Profile of the rural surgeon. PMID- 18506527 TI - Simplex PCR assay for positive identification of genetic sex in the Japanese medaka, Oryzias latipes. AB - The medaka, Oryzias latipes, is a very popular model in biomedical research, particularly for elucidating sex differentiation and determination mechanisms and effects of endocrine disruptors among others. These studies require a sensitive, accurate, rapid, and reliable technique for genetic sexing of eggs, larvae, and adults. In this study, we report a simplex polymerase chain reaction approach that uses a single pair of primers for simultaneous amplification of sex-specific amplicons. Males and females yield a single diagnostic band of 933 and 1,906 bp, respectively, in three different strains of medaka tested, permitting gender identification accurately of both immature and adult fish. This technique will be useful in both ecological and biomedical researches that employ medaka and rely on genetic sexing. PMID- 18506529 TI - Hemorrhoids can be a source of obscure gastrointestinal bleeding that requires transfusion: report of five patients. AB - PURPOSE: Hemorrhoids have been rarely reported to be a source of transfusion dependent, obscure gastrointestinal bleeding. We report the diagnosis and management of a series of patients in whom hemorrhoids were the source of obscure gastrointestinal bleeding that was severe enough to require transfusion. METHODS: Five patients, who presented with severe hematochezia or obscure gastrointestinal bleeding, during a 24-month period had had an extensive workup for gastrointestinal bleeding. All had required transfusion of 2 units or more of blood. In the absence of other causes of bleeding, the five patients had unprepared flexible sigmoidoscopy on the same day that they reported hematochezia to exclude a proximal source of bleeding. All were found to have internal hemorrhoids as a likely source of bleeding, which was confirmed at exploration under anesthesia, and were treated by surgical hemorrhoidectomy. RESULTS: The five patients underwent surgical, Ferguson-type, hemorrhoidectomy. One patient required 1 unit of blood immediately postoperatively, but none have required further transfusion or reported hematochezia in more than three months of follow up. CONCLUSIONS: We have shown that hemorrhoids can be a source of transfusion dependent, obscure gastrointestinal bleeding. This aspect of the common problem of hemorrhoidal bleeding has not been previously reported, which reflects underreporting or missed diagnosis. PMID- 18506530 TI - An unexpected mixture of substances in the defensive secretions of the tubuliferan thrips, Callococcithrips fuscipennis (Moulton). AB - Adults and larvae of the thrips Callococcithrips fuscipennis (Moulton) (Thysanoptera: Tubulifera: Phlaeothripidae) live in the sticky wax masses of adult females of the felt scale insect Callococcus acaciae (Maskell) (Sternorrhyncha: Coccoidea: Eriococcidae). The scale is sessile and feeds on Kunzea shrubs (Myrtales: Myrtaceae). If stressed, the thrips produce droplets of secretions. The mixture contains pentadecane, tridecane, two monoterpenoids, hexadecyl butanoate, and smaller amounts of 15 other esters of long-chain unbranched alcohols identified as acetates, butanoates, hexanoates, and octanoates. The monoterpenoids are dolichodial, an iridoid, and an unknown substance with a mass spectrum very similar to that of anisomorphal and peruphasmal, diastereomers of dolichodial, but with a different retention time. Iridoids, butanoates, hexanoates, and octanoates have not been previously identified in Thysanoptera. PMID- 18506528 TI - Functional prestin transduction of immature outer hair cells from normal and prestin-null mice. AB - Prestin is a membrane protein in the outer hair cell (OHC) that has been shown to be essential for electromotility. OHCs from prestin-null mice do not express prestin, do not have a nonlinear capacitance (the electrical signature of electromotility), and are smaller in size than wild-type OHCs. We sought to determine whether prestin-null OHCs can be transduced to incorporate functional prestin protein in a normal fashion. A recombinant helper-dependent adenovirus expressing prestin and green fluorescent protein (HDAd-prestin-GFP) was created and tested in human embryonic kidney cells (HEK cells). Transduced HEK cells demonstrated membrane expression of prestin and nonlinear capacitance. HDAd prestin-GFP was then applied to cochlear sensory epithelium explants harvested from wild-type and prestin-null mice at postnatal days 2-3, the age at which native prestin is just beginning to become functional in wild-type mice. At postnatal days 4-5, we investigated transduced OHCs for (1) their prestin expression pattern as revealed by immunofluorescence; (2) their cell surface area as measured by linear capacitance; and (3) their prestin function as indicated by nonlinear capacitance. HDAd-prestin-GFP efficiently transduced OHCs of both genotypes and prestin protein localized to the plasma membrane. Whole-cell voltage clamp studies revealed a nonlinear capacitance in transduced wild-type and prestin-null OHCs, but not in non-transduced cells of either genotype. Prestin transduction did not increase the linear capacitance (cell surface area) for either genotype. In peak nonlinear capacitance, voltage at peak nonlinear capacitance, charge density of the nonlinear capacitance, and shape of the voltage-capacitance curves, the transduced cells of the two genotypes resembled each other and previously reported data from adult wild-type mouse OHCs. Thus, prestin introduced into prestin-deficient OHCs segregates normally to the cell membrane and generates a normal nonlinear capacitance, indicative of normal prestin function. PMID- 18506531 TI - Distance communication of sexual status in the crayfish Orconectes quinebaugensis: female sexual history mediates male and female behavior. AB - Chemical communication plays an important role in mediating social interactions of many taxa, particularly arthropods. Many individuals communicate information about their reproductive status to potential mates through distance and/or contact pheromones, an ability that may be advantageous to both signalers and receivers. In this paper, we describe tests of two hypotheses on the role of distance communication in the reproductive behaviors of crayfish (Orconectes quinebaugensis). First, we hypothesized that male crayfish would show stronger attraction towards virgin females (females with no viable sperm) than towards non virgin females because of the fitness costs (to males) associated with sperm competition. Second, we hypothesized that female crayfish should show differential responses to mature male signals depending on their own sexual history: virgin females should be more strongly attracted to male signals than should non-virgin females because they must mate at least once to be able to fertilize eggs in the spring. Data from two Y-maze experiments yielded support for both hypotheses: males were attracted to signals from virgin females, but not to signals from non-virgins. Likewise, virgin females were attracted to signals from males, but non-virgin females were not. We discuss our data in the context of the potential costs and benefits of mate searching and suggest that distance chemical communication of sexual status may be particularly advantageous when the costs of mate searching are high. PMID- 18506533 TI - When does skin excision allow the achievement of an adequate local control rate in patients with squamous cell carcinoma involving the buccal mucosa? AB - BACKGROUND: The role of cheek skin excision in patients with squamous cell carcinoma of the buccal mucosa (BSCC) remains controversial. We sought to investigate when skin excision is needed to achieve an adequate local control. METHODS: A total of 331 patients with BSCC were reviewed. Skin preservation was pursued when the distance between the tumor and the skin as determined by imaging was >or=13 mm (1 cm surgical margin plus 0.3 cm skin preservation). Two hundred and thirty patients (69.5%) underwent skin excision. Postoperative adjuvant radiotherapy (n = 182) was performed in patients with pathological T4 disease, metastases in cervical lymph nodes or close pathological margins ( or = 9 visits, P < 0.001), lower costs ($24,400 vs. $23,400, P < 0.05), less in-hospital death (44% vs. 40%, P < 0.01), and fewer preventable hospitalizations for those with congestive heart failure (adjusted odds ratio, aOR = 0.82, P < 0.001) and chronic obstructive pulmonary disease (aOR = 0.81, P = 0.02). CONCLUSIONS: Primary care visits in the preceding year are associated with less, and less costly, end-of-life hospital utilization. Increased primary care access for Medicare beneficiaries may decrease costs and improve quality at the end of life. PMID- 18506546 TI - Preferences for human papillomavirus testing with routine cervical cancer screening in diverse older women. AB - BACKGROUND: Human papillomavirus (HPV) testing is increasingly being used to determine the optimal cervical cancer screening interval in older women. Little is known about women's attitudes toward HPV testing or how these attitudes may influence medical discussions about cervical cancer screening. METHODS: Preferences for HPV and concomitant Papanicolaou (Pap) testing were assessed through in-person interviews with diverse women aged 50 to 80 years recruited from community and university-based practices. RESULTS: Eight hundred and sixty five women (257 White, 87 African American, 149 Latina, and 372 Asian) were interviewed. Approximately 60% of participants wanted to be tested for HPV and another 15% would undergo testing if recommended by their physician. Among those wanting HPV testing, 94% would want more frequent than annual Pap tests if they had a positive HPV test and a normal Pap test. Two thirds of those under age 65 would be willing to switch to triennial Pap testing, and half of those aged 65 and older would be willing to discontinue Pap testing, if they had a negative HPV test and normal Pap test. Preferences for testing varied by ethnicity, age, place of birth, and cancer history. CONCLUSIONS: The majority of older women were willing to use HPV testing to make decisions about frequency and duration of cervical cancer screening, but up to one third would want at least annual, ongoing screening regardless of HPV test results. Efforts should be made to ensure that HPV testing is used to reinforce appropriate utilization of screening tests. PMID- 18506547 TI - Autoimmune pancreatitis and concurrent small lymphocytic lymphoma: not just a coincidence? AB - CASE: A 76-year-old gentleman presented with painless jaundice, weight loss, and anorexia. Computed tomography imaging revealed fullness of the pancreatic head and multiple enlarged retroperitoneal lymph nodes. Cholangiogram revealed a distal common bile duct stricture. Due to concerns of malignancy, the patient underwent operative exploration. Several enlarged lymph nodes in the aortocaval region and a firm hard mass in the pancreatic head were found. Frozen section from one of the lymph nodes was suspicious for low-grade lymphoma. A pancreaticoduodenectomy was performed. Histologic analysis of the pancreatic head revealed a lymphoplasmacytic infiltrate with stromal fibrosis consistent with autoimmune pancreatitis. The retroperitoneal lymph nodes were involved by small lymphocytic lymphoma. DISCUSSION: Autoimmune pancreatitis is the most common benign diagnosis after pancreatic resection for presumed malignancy. It has a well-documented association with autoimmune conditions, such as Sjogren's syndrome, inflammatory bowel disease, and sclerosing cholangitis. Additionally, chronic lymphocytic leukemia-small lymphocytic lymphoma is often associated with autoimmune phenomena, most notably autoimmune hemolytic anemia. However, an association between autoimmune pancreatitis and small lymphocytic lymphoma has not been previously described. To our knowledge, this is the first reported case of a patient with concurrent autoimmune pancreatitis and small lymphocytic lymphoma. PMID- 18506548 TI - Staged laparoscopic sleeve gastrectomy followed by Roux-en-Y gastric bypass for morbidly obese patients: a risk reduction strategy. AB - BACKGROUND: In our centre laparoscopic Roux-en-Y gastric bypass (LRYGBP) is the most effective weight loss surgical procedure performed. However, LRYGBP may be associated with higher risk of peri- and postoperative complications in contrast to a purely restrictive procedure to justify this procedure on all comers. Laparoscopic sleeve gastrectomy (LSG) as a staged procedure may be an alternate risk reduction strategy. The aim of this study is to report on the short-term outcomes of LSG, the effect on operative risk reduction and resolution of comorbidities. METHODS: A prospective review of 138 patients who underwent consecutive LSG from November 2004 to November 2006 was performed. Data were collected on all patients who attended the three to six monthly clinical follow up and/or the patient questionnaire. Data collection included demographics, degree of weight reduction, postoperative complications, and changes in comorbidities. RESULTS: Median BMI was 50.60 kg/m(2) (33-82). Of the patients, 46.38% had a BMI >or=50 kg/m(2). The overall median postoperative excess weight loss (EWL) was 43.26%, 31.08% at 6 months, 54.50% at 12 months, 51.47% at 18 months and 46.05% at 24 months. Of the patients, 39% had resolution of type 2 diabetes mellitus, 48% had resolution of dyslipidemia, 29% in hypertension, 52% in obstructive sleep apnea. Complication rate was 5.07% and four patients needed further surgical intervention. The mortality rate was zero. CONCLUSION: LSG does minimize postoperative complication rates significantly on high-risk patients and achieves effective short-term weight loss with resolutions in comorbidities. Additional studies are required to evaluate LSG as a stand-lone procedure. PMID- 18506549 TI - Endoscopic cervical bariatric surgery: follow-up study in a porcine model. AB - BACKGROUND: The realisation of bariatric surgery has to date modified the digestive process solely through procedures within the abdominal cavity. However, endocrine surgeons have recently demonstrated the feasibility of a minimally invasive approach to the neck. In this study, we explored the feasibility, safety and weight progression of a bariatric procedure performed at the neck. METHODS: Eleven 40-50 kg Yorkshire pigs underwent endoscopic placement of an adjustable band to the cervical esophagus (ECB). Weight was monitored at postoperative days 15, 30, and after 7 weeks; weight progression was compared with an identical group of pigs who underwent a sham procedure. At autopsy, the surgical site was evaluated in a microscopic and macroscopic manner. RESULTS: Mean operating time was 66 +/- 5.76 min. All pigs tolerated the procedure well, except one subject that experienced food intolerance. The ECB group experienced significantly slower weight gain than the sham group (P = 0.005). Proper location of the band and absence of microscopic lesions at the esophageal wall were confirmed at autopsy and pathological examination. CONCLUSION: Bariatric surgery at the neck is feasible and produces effects on weight reduction. Further refinements and longer observation periods are required to propose this procedure as safe and effective alternative in humans. PMID- 18506550 TI - Adaptive and neuroendocrine procedures: a new pathway in bariatric and metabolic surgery. PMID- 18506551 TI - Carcinoid tumors and morbid obesity. AB - Carcinoid is a rare gastrointestinal tumor, with an incidence varying from 1 to 2.5 per 100,000 in the general population. In this article, we report an elevated incidence of carcinoid tumor in an obese population, showing the importance of performing an endoscopic procedure before bariatric surgery. PMID- 18506552 TI - Laparoscopic sleeve gastrectomy in ethnic obese Chinese. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness and safety of laparoscopic sleeve gastrectomy (LSG) for the treatment of obesity in ethnic Chinese in Hong Kong. METHODS: Seventy consecutive Chinese patients (49 females; mean age 34.7+/-8.8 [range 18-56] years) received LSG for the treatment of obesity from May 2006 to Nov 2007 as a stand-alone procedure for weight reduction. Mean baseline body weight (BW) and body mass index (BMI) were 108.9+/ 22.1 kg (range 71.0-164.9 kg) and 40.7+/-7.8 kg/m(2) (range 27.4-68.4 kg/m(2)), respectively. Outcome measures were collected and assessed in a prospective manner. RESULTS: All procedures were performed laparoscopically with no conversion. There was neither mortality nor any postoperative complications that required reoperation. Major complication occurred in two patients (2.9%; esophagogastric junction [EGJ] leak and stomach tube stricture). Mean follow-up was 7.1+/-5.0 months. Mean procedure time was 90.6+/-39.4 min, and mean hospital stay was 3.8+/-2.3 days. Mean BMI loss was 6.3+/-2.5, 9.0+/-3.4 and 12.3+/-4.5 kg/m(2) at 3, 6, and 12 months. Mean percent of excess BW loss was 48.5+/-28.4, 69.7+/-31.7, and 63.5+/-29.4 at 3, 6, and 12 months. CONCLUSION: LSG is safe and effective in achieving significant weight loss in obese ethnic Chinese patients. PMID- 18506553 TI - Relationship between knee anthropometry and surgical time in total knee arthroplasty in severely and morbidly obese patients: a new prognostic index of surgical difficulty. AB - BACKGROUND: Total knee arthroplasty (TKA) in patients with severe and morbid obesity is one of the current challenges in prosthetic knee surgery. The body mass index (BMI) is used to identify patients who may present difficulties during surgery and postoperative complications. We carried out a prospective study with an initial hypothesis that BMI is not associated with tourniquet time in obese patients undergoing TKA and that some anthropometric parameters may be useful in predicting tourniquet time in severely and morbidly obese patients. METHODS: One hundred consecutive patients diagnosed with knee osteoarthritis with BMI > or =35 kg/m(2) scheduled for TKA were prospectively studied. Suprapatellar, infrapatellar, and supra/infrapatellar anthropometric indexes were calculated before surgery. The tourniquet time was determined. RESULTS: The mean BMI was 39.81 kg/m(2) (SD +/- 3.75). A total of 58% of patients were classified as class III obesity (BMI 35-39.99) and 42% as class IV (BMI > or = 40) Mean tourniquet time was 41.67 min (SD +/- 9.26). There was no association between the BMI and tourniquet time. The suprapatellar index was negatively associated with tourniquet time (p < 0.038). DISCUSSION: The BMI is not the only parameter that should be considered in order to identify severely and morbidly obese patients who may have more surgical difficulties during TKA. Preoperative determination of the suprapatellar index helped us to classify these patients according to the morphology of the knee and predicted a longer tourniquet time and, therefore, greater surgical difficulty, in patients with a suprapatellar ratio below 1.6 in this study. PMID- 18506554 TI - Hepatic evisceration after cholecystectomy in a superobese patient. AB - Gallbladder pathology, in general, and cholelithiasis, in particular, are more common in the morbidly obese. Obesity is a risk factor for conversion to open surgery in laparoscopic cholecystectomy. Obesity is also a risk factor for evisceration after laparotomy in adults. Hepatic evisceration after cholecystectomy is rare. We describe a case of right liver lobe evisceration diagnosed by abdominal computed tomography in a superobese patient (body mass index 57 kg/m(2)) after emergency laparoscopic surgery for acute calculous cholecystitis converted to open surgery. PMID- 18506555 TI - Small increase of actual physical activity 6 months after total hip or knee arthroplasty. AB - Limitation in daily physical activity is one of the reasons for total hip arthroplasty (THA) or total knee arthroplasty (TKA). However, studies of the effects of THA or TKA generally do not determine actual daily activity as part of physical functioning. We determined the effect of THA or TKA on patients' actual physical activity and body function (pain, stiffness), capacity to perform tasks, and self-reported physical functioning. We also assessed whether there are differences in the effect of the surgery between patients undergoing THA or TKA and whether the improvements vary between these different outcome measures. We recruited patients with long-standing end-stage osteoarthritis of the hip or knee awaiting THA or TKA. Measurements were performed before surgery and 3 and 6 months after surgery. Actual physical activity improved by 0.7%. Patients' body function, capacity, and self-reported physical functioning also improved. The effects of the surgery on these aspects of physical functioning were similar for THA and TKA. The effect on actual physical activity (8%) was smaller than on body function (80%-167%), capacity (19%-36%), and self-reported physical functioning (87%-112%). Therefore, in contrast to the large effect on pain and stiffness, patients' capacity, and their self-reported physical functioning, the improvement in actual physical activity of our patients was less than expected 6 months after surgery. LEVEL OF EVIDENCE: Level I, prospective study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18506556 TI - Fas death pathway in sarcomas correlates with epidermal growth factor transcription. AB - Modulation of apoptosis may influence sarcoma pathogenesis and/or aggressiveness. The Fas death pathway, mediated by FasL or TGFbeta, is one of two apoptotic pathways. Recent studies report that EGF can modulate TGFbeta and/or FasL expression/activity; thus, EGF has the potential to influence activation of the Fas pathway. EGF is not always detectable in mesenchymal tumors; therefore, we hypothesized EGF would define which Fas ligand predominates. We assayed 57 surgically removed human sarcomas for 10 genes involved in the Fas pathway. Skeletal muscle biopsies from eight patients served as controls. Sample transcripts were detected by real-time RT-PCR. We attempted to identify relevant predictor variables. The 57 sarcomas were segregated into two categories defined by EGF mRNA content: (1) 23 tumors with EGF concentrations that approximated muscle EGF transcript levels (high-EGF tumors); and (2) 34 tumors that either lacked EGF mRNA, or whose mRNA levels were very low and frequently undetected by PCR (low-EGF tumors). TGFbeta1 expression best predicted Fas transcript concentrations in the 34 low-EGF sarcomas, while FasL predicted Fas mRNA levels in the remaining 23 high-EGF sarcomas. The results suggest ligand activity in the Fas death pathway correlates with EGF transcription in sarcomas. PMID- 18506557 TI - Increased carrying angle is a risk factor for nontraumatic ulnar neuropathy at the elbow. AB - The literature suggests a possible relationship between carrying angle and nontrauma-related ulnar neuropathy. To confirm that relationship, we asked whether carrying angle is a risk factor in patients with nontrauma-related ulnar neuropathy. We measured the carrying angles of the elbow in 36 patients with a clinically and electrophysiologically confirmed diagnosis of nontraumatic ulnar neuropathy at the elbow and in 50 healthy control subjects. Correlation analysis was performed between carrying angles and parameters of nerve conduction studies, including nerve conduction velocities and amplitudes of muscle and nerve action potentials. The mean carrying angle was greater in the patients than in the control subjects. Females had a greater carrying angle than males. We observed an inverse relationship between carrying angles and motor nerve conduction velocities at cross-elbow segments of the ulnar nerves and with sensory nerve conduction velocities of the distal ulnar nerves. An increased carrying angle of the elbow appears to be an independent risk factor of nontrauma-related ulnar neuropathy. LEVEL OF EVIDENCE: Level III, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18506558 TI - Chondrocyte apoptosis: implications for osteochondral allograft transplantation. AB - Osteochondral allograft transplantation is a useful technique to manage larger articular cartilage injuries. One factor that may compromise the effectiveness of this procedure is chondrocyte cell death that occurs during the storage, preparation, and implantation of the osteochondral grafts. Loss of viable chondrocytes may negatively affect osteochondral edge integration and long-term function. A better understanding of the mechanisms responsible for chondrocyte loss could lead to interventions designed to decrease cell death and improve results. Recent studies indicate that apoptosis, or programmed cell death, is responsible for much of the chondrocyte death associated with osteochondral allograft transplantation. Theoretically, some of these cells can be rescued by blocking important apoptotic mediators. We review the role of apoptosis in cartilage degeneration, focusing on apoptosis associated with osteochondral transplantation. We also review the pathways thought to be responsible for regulating chondrocyte apoptosis, as well as experiments testing inhibitors of the apoptotic pathway. These data suggest that key contributors to the apoptotic process can be manipulated to enhance chondrocyte survival. This knowledge may lead to better surgical outcomes for osteochondral transplantation. PMID- 18506559 TI - Allograft reconstruction after sarcoma resection in children younger than 10 years old. AB - Preservation of limb function in pediatric oncology patients is challenging with the ongoing growth of limbs contralateral to reconstructed limbs. We analyzed 22 patients younger than 10 years old who received an allograft after resection of a bone sarcoma with a minimum followup of 2 years (mean, 4 years; range, 2-14 years). The mean age was 7 years (range, 2-10 years). There were 16 boys and six girls with 17 osteosarcomas and five Ewing's sarcomas. Thirteen reconstructions were performed with an intercalary allograft and nine with an osteoarticular allograft. Physes were uninvolved in five patients and one physis in 17. We documented outcomes using the Musculoskeletal Tumor Society functional and the International Society of Limb Salvage radiographic scoring systems. At last followup, three of the 22 patients died of their tumor, one was alive but with an amputation, and 18 retained their limbs. These 18 patients had an average functional score of 27 points and a mean radiographic score of 94%. Eight complications required a second surgery; in four, the allograft was removed (one infection, one local recurrence, two fractures) and in four, the allograft was preserved (two local recurrences, one fracture, one nonunion). We consider biologic reconstruction with allografts after sarcoma resection an appropriate reconstructive procedure in young children. LEVEL OF EVIDENCE: Level IV, therapeutic study. PMID- 18506560 TI - Enhancing osteochondral allograft viability: effects of storage media composition. AB - Osteochondral allograft transplantation is a well-accepted treatment for articular cartilage damage. However, chondrocyte viability declines during graft storage, which may compromise graft performance. We first tested the hypothesis that the composition of commonly used storage media affects the viability of articular chondrocytes over time; we then tested the hypothesis that the addition of insulin growth factor-1 or the apoptosis inhibitor ZVAD-fmk could enhance the storage properties of serum-free media. Bovine osteochondral grafts were stored at 4 degrees C in lactated Ringer's, Dulbecco's modified eagle's media (DMEM), DMEM supplemented with either insulin growth factor-1 or ZVAD-fmk, and a commercial storage media. Chondrocyte viability in lactated Ringer's declined rapidly to 20.4% at 2 weeks. Viability in DMEM declined more slowly to 54.8% at 2 weeks and 31.2% at 3 weeks. Viability in commercial storage media was 83.6% at 3 weeks and 44.8% at 4 weeks. Viability was increased in DMEM + insulin growth factor-1 (56.4%) and DMEM + ZVAD (52.4%) at 3 weeks compared with DMEM alone. These results confirm the hypotheses that media composition greatly influences chondrocyte viability during cold storage and that insulin growth factor-1 and ZVAD improve the storage properties of DMEM. PMID- 18506561 TI - The Achilles tendon insertion is crescent-shaped: an in vitro anatomic investigation. AB - Anatomic and operative textbooks and current literature do not clearly describe the Achilles tendon interface to the calcaneal tuberosity. We dissected 51 specimens to identify the detailed anatomy of the Achilles tendon insertion. Achilles tendon fascicles expanded from the anterior aspect of the distal Achilles tendon over the retrocalcaneal bursa to the anterior part of the Haglund's tuberosity in nearly half of the specimens. The insertion of the transverse section of the Achilles tendon regularly had a crescent-shape corresponding to the posterior calcaneal prominence. In transverse sections, all specimens had a curved appearance with a radius of curvature ranging from 13.8 mm to 43.6 mm (mean, 20.4 mm) and Achilles tendon extensions to the lateral and medial calcaneal surfaces reached 1.0 mm (mean) and 3.5 mm (mean) anterior in relation to the most posterior point of the calcaneal tuberosity. Knowledge of the arcuate configuration and of the medial and lateral extensions of the plantaris and the Achilles tendon insertion with respect to the transverse plane is important to avoid iatrogenic complications during resection of Haglund's tuberosity. PMID- 18506562 TI - Gene therapy to enhance allograft incorporation after host tissue irradiation. AB - Structural bone allografts are used to reconstruct large skeletal defects after tumor surgery. Although allograft-related complications are declining, the use of perioperative radiation therapy is associated with a poorer outcome. Recently, BMP-2 levels in the host bed were reportedly diminished after exposure to radiation doses consistent with those used perioperatively to treat musculoskeletal sarcoma. Reintroduction of this osteogenic protein may circumvent the deleterious effects of preoperative radiation on allograft incorporation. We introduced a novel polymeric BMP-2 gene delivery system into the host-allograft junctions at the time of transplantation in an ovine tibial defect model with or without preoperative exposure to 50 Gy radiation. After 4 months, we noted no radiographic or histologic improvements in allograft incorporation after preoperative radiation and BMP-2 reintroduction; however, 50 Gy radiation was associated with increased porosity in the interface regions and poorer radiographic healing. We identified no BMP2-expressing cells or protein in the interface at the study end point, suggesting the polymeric gene delivery system was unable to promote extended expression of the protein or induce a healing response. Although gene therapy may hold promise as a novel technique to improve allograft incorporation, our data do not support that contention with the current approach. PMID- 18506563 TI - Imaging analysis of the in vivo bioreactor: a preliminary study. AB - The in vivo bioreactor is a hermetically sealed, acellular hydroxyapatite scaffold coated with growth factors that has a pulsating vascular pedicle leash threaded through its center. Tissue-engineered bone is created in weeks while the bioreactor remains embedded under the skin of an animal. The bioreactor also provides a model to study osteogenesis and pathologic scenarios such as tumor progression and metastasis by creating a controlled microenvironment that makes skeletogenesis amenable to genetic and physical manipulation. Animal euthanasia is required to quantitate bioreactor osteogenesis through histomorphometry. Nondestructive measures of new bone growth within the bioreactor are critical to future applications and are the primary questions posed in this study. We compared microcomputed tomography and micro-MRI assessments of bioreactor osteogenesis with conventional histomorphometric measurements in 24 bioreactors and asked if new bone formation could be calculated while the animal was alive. Microcomputed tomography visually, but not numerically, differentiated engineered new bone on its coral scaffold. Dynamic contrast-enhanced micro-MRI demonstrated augmented vascular flow through the bioreactor. Three-dimensional imaging can nondestructively detect tissue-engineered osteogenesis within the implanted bioreactor in vivo, furthering the usefulness of this unique model system. PMID- 18506564 TI - HLA sensitization and allograft bone graft incorporation. AB - Achieving union between host bone and massive structural allografts can be difficult. Donor and recipient human leukocyte antigen (HLA) mismatches and recipient antibody response to donor HLA antigens might affect union. In a prospective multiinstitutional study, we enrolled a consecutive series of patients receiving cortex-replacing, massive structural bone allografts to determine the rate of donor-specific HLA antibody sensitization and to investigate the potential effect of such HLA alloantibody sensitization on allograft incorporation. HLA typing of patients and donors was determined by molecular typing methods. Donor-specific HLA sensitization occurred in 57% of the patients but had no demonstrable effect on graft incorporation or union. The type of host-allograft junction did have a major effect on graft incorporation. Cortical-to-cortical allograft-to-host junctions healed more slowly (mean, 542 days) than corticocancellous to corticocancellous allograft-to-host junctions (mean, 243 days). Although HLA sensitization does not appear to delay structural allograft bone incorporation, further followup is required to determine if there is an association between HLA sensitization and long-term graft survival. Based on these preliminary data, measures to further minimize or modulate HLA sensitization or response are not indicated at present for the purposes of improving structural bone allograft union. LEVEL OF EVIDENCE: Level II, prognostic study. PMID- 18506565 TI - Shaped, stratified, scaffold-free grafts for articular cartilage defects. AB - One goal of treatment for large articular cartilage defects is to restore the anatomic contour of the joint with tissue having a structure similar to native cartilage. Shaped and stratified cartilaginous tissue may be fabricated into a suitable graft to achieve such restoration. We asked if scaffold-free cartilaginous constructs, anatomically shaped and targeting spherically-shaped hips, can be created using a molding technique and if biomimetic stratification of the shaped constructs can be achieved with appropriate superficial and middle/deep zone chondrocyte subpopulations. The shaped, scaffold-free constructs were formed from the alginate-released bovine calf chondrocytes with shaping on one (saucer), two (cup), or neither (disk) surfaces. The saucer and cup constructs had shapes distinguishable quantitatively (radius of curvature of 5.5 +/- 0.1 mm for saucer and 2.8 +/- 0.1 mm for cup) and had no adverse effects on the glycosaminoglycan and collagen contents and their distribution in the constructs as assessed by biochemical assays and histology, respectively. Biomimetic stratification of chondrocyte subpopulations in saucer- and cup-shaped constructs was confirmed and quantified using fluorescence microscopy and image analysis. This shaping method, combined with biomimetic stratification, has the potential to create anatomically contoured large cartilaginous constructs. PMID- 18506566 TI - Multiple brain calcifications in a patient with systemic lupus erythematosus. AB - We describe a rare case of severe neuropsychiatric systemic lupus erythematosus with cerebral atrophy and multiple paraventricular, basal ganglia, cortex, cerebral white matter, and cerebellum calcifications detected on cerebral CT. These calcifications are probably secondary to necrosis focus of repeated episodes of vessels inflammation. PMID- 18506567 TI - Safety profile of tacrolimus in patients with rheumatoid arthritis. AB - We assessed the safety of tacrolimus therapy for rheumatoid arthritis. Forty-two patients who started tacrolimus therapy between April 2005 and July 2006 were investigated retrospectively using data from their medical records up to June 2007. The cumulative treatment continuation rate was assessed by the Kaplan-Meier method. Fisher's exact test was used to compare gastrointestinal symptoms between different tacrolimus doses and between the presence and absence of each concomitant medication. The mean (+/-SD) observation period was 288 +/- 238 days. The cumulative treatment continuation rate was, respectively, 59.5% and 38.1% at 6 months and 1 year after the patients started treatment. Tacrolimus was discontinued in 28 patients, and was discontinued because of adverse reactions in 21 patients. Gastrointestinal symptoms were the most common adverse reactions (45.2% = 19/42 patients), followed by infections and hyperglycemia. Tacrolimus was discontinued in 9/19 patients with gastrointestinal symptoms, and was discontinued within 60 days of starting treatment in seven of them. Nausea and vomiting led to discontinuation in seven patients (within 60 days of starting treatment in six of them). The incidence of gastrointestinal symptoms was higher in patients receiving a daily dose >or=2 mg than in those receiving <2 mg/day. During treatment of rheumatoid arthritis by oral tacrolimus therapy, gastrointestinal symptoms were common, early, and dose-dependent. However, these symptoms were not severe and did not cause any serious safety problems. PMID- 18506568 TI - Melioidosis--an unusual cause of septic arthritis. AB - Melioidosis is an infection caused by Burkholderia pseudomallei. It is an important human pathogen in the tropical area. The clinical manifestations are protean with multisystem involvement. Septic arthritis and prostatic abscess are rare but well-recognized forms of the disease. Herein we report a case of melioidosis presenting with a rare combination of septic arthritis, prostatic abscess, and septicemia. PMID- 18506569 TI - Hyperimmunoglobulinemia D and periodic fever syndrome; treatment with etanercept and follow-up. AB - The hyperimmunoglobulinemia D and periodic fever syndrome (HIDS) is an autoinflammatory syndrome. It is caused by the mutations of the mevalonate kinase gene. There is no consensus for specific therapy of HIDS, but there are some case reports and studies in regards to its treatment with drugs like colchicine, steroids, nonsteroid anti-inflammatory drugs, simvastatin, anakinra, thalidomide, and etanercept. We are reporting a case evaluated for the complaints of abdominal pain and febrile episodes with massive hepatomegaly, not common finding on physical examination, its treatment with etanercept, and long-term follow-up. PMID- 18506570 TI - Spinal cord atrophy in spinocerebellar ataxia type 3 and 6 : impact on clinical disability. AB - OBJECTIVE: To quantify spinal cord atrophy and its impact on clinical disability in spinocerebellar ataxia (SCA) type 3 and 6. METHODS: Atrophy of the upper spinal cord was assessed by high resolution T1-weighted MRI of patients with SCA3 (n = 14) and SCA6 (n = 10). Furthermore, two groups of age- and sex-matched healthy control subjects (n = 24,) corresponding to the two SCA groups, were studied. Images were post-processed by a semi-automated volumetry method combining a marker based segmentation and an automatic histogram method facilitating highly reliable quantification and morphometry of the upper cervical cord in vivo. RESULTS: We found a significant reduction of normalized mean crosssectional area of the spinal cord in SCA3 (p < 0.0005), whereas in SCA6 patients normalized mean crosssectional area was in the normal range (p = 0.379). No correlation was found between spinal cord atrophy and disease duration as well as CAG repeat length in both subtypes. In SCA6 a negative dependency between clinical disability, as expressed by the International Cooperative Ataxia Rating Scale as a well established ataxia score, and the mean cross-sectional area was found (p = 0.02). A similar correlation was observed in SCA3 but did not reach statistical significance. CONCLUSION: Our results quantify for the first time in vivo spinal cord atrophy as a non-cerebellar neurodegenerative process in SCA3. Our results suggest MR volumetry of the upper cervical cord as a marker of functional importance in SCA3 and SCA6. PMID- 18506571 TI - High-mobility group protein A1 binds herpes simplex virus gene regulatory sequences and affects their expression. AB - The high-mobility group protein A1 (HMGA1), which regulates mammalian gene expression by altering chromatin architecture, was found to bind at multiple sites within the promoter regions of all of the herpes simplex virus type 1 (HSV 1) immediate early genes, as well as a representative early (tk) gene and one late (gC) gene, both in vitro and in vivo. Infected cell polypeptide (ICP) 4, the major HSV-1 regulatory protein, binds these promoters both in vitro and in vivo, and HMGA1 enhances its in vitro binding. In transient expression experiments, HMGA1 modified the effects of both ICP4 and ICP0, another virus transactivator, on virus gene expression in a promoter-specific manner, but it had no effect on the transactivation of immediate-early promoters by VP16. These data indicate that host-cell architectural chromatin proteins could influence the interactions of host-cell and viral transcription factors with the virus DNA regulatory elements and affect HSV-1 gene expression. PMID- 18506572 TI - Genome comparison of virulent and avirulent strains of the Pichinde arenavirus. AB - A virulent (P18) strain of the Pichinde arenavirus produces a disease in guinea pigs that somewhat mimics human Lassa fever, whereas an avirulent (P2) strain of this virus is attenuated in infected animals. It has been speculated that the composition of viral genomes may confer the degree of virulence in an infected host; the complete sequence of the viral genomes, however, is not known. Here, we provide for the first time genomic sequences of the S and L segments for both the P2 and P18 strains. Sequence comparisons identify three mutations in the GP1 subunit of the viral glycoprotein, one in the nucleoprotein NP, and five in the viral RNA polymerase L protein. These mutations, alone or in combination, may contribute to the acquired virulence of Pichinde virus infection in animals. The three amino acid changes in the variable region of the GP1 glycoprotein subunit may affect viral entry by altering its receptor-binding activity. While NP has previously been shown to modulate host immune responses to viral infection, we found that the R374 K change in this protein does not affect the NP function of suppressing interferon-beta expression. Four out of the five amino acid changes in the L protein occur in a small region of the protein that may contribute to viral virulence by enhancing its function in viral genomic RNA synthesis. PMID- 18506573 TI - Recruiting Latino women in the U.S. and women in Mexico in postpartum depression prevention research. AB - The negative consequences of untreated depression on the health and well-being of women and their children are well-documented, underscoring the need to develop effective interventions to prevent the onset of major depression during the perinatal period. This article describes recruitment data from two randomized controlled trials of preventive interventions for postpartum depression: one conducted with immigrant Latinas in Washington, DC, United States, and the other with women in Mexico City, Mexico. In both countries, pregnant women met a priori eligibility criteria and were randomized into an 8-week theory-based group intervention. Two hundred and seventeen Latinas in the U.S. and 377 women born and raised in Mexico were enrolled in their respective countries. The recruitment rates (i.e., the number of participants who met eligibility criteria, consented, and randomized into the study) were 70% in the U.S. and in Mexico. Issues and recommendations related to recruiting Hispanic women into preventive intervention trials for postpartum depression are discussed. PMID- 18506574 TI - An audit of pregnant women with severe mental illness referred during the first 2 years of a new perinatal mental health service. AB - The importance of identifying pregnant women at high risk of postpartum psychotic illness has been highlighted by recent enquiries and guidelines. It has been recommended that these women are referred to perinatal mental health services, and that individualised care plans are made prior to delivery. This audit describes a cohort of 45 women, referred to a new perinatal mental health service, with a history of psychotic illness or at risk of developing a postpartum psychotic episode. It describes the characteristics of this group, and the outcomes in terms of relapse and whether their children remained in their care. Thirty-nine women (87%) were seen for assessment. Sixty-seven per cent were under the care of a community mental health team. Twenty-one (53.8%) were well at the time of referral. Planned pregnancies were uncommon (15%). All women seen antenatally had care plans written to address the high risk of postpartum relapse. Ten (26%) women had psychiatric admissions during pregnancy. Fifteen (38%) relapsed or had admissions during the first postpartum year. At 1 year postpartum, 74% women had care of their children; 18% children had been adopted or were in foster care; and the outcome was unknown for 8%. PMID- 18506575 TI - Family caregiver role and premenstrual syndrome as associated factors for postnatal depression. AB - The goal of this study was to identify sociodemographic, psychopathological, and obstetric risk factors associated with postnatal depression (PND) and their relative weight. A cross-sectional two-stage design was used. All consecutive women receiving a routine check-up 6 weeks postpartum at Obstetric Services during a 1-year period were included. In the first stage, women completed the Edinburgh Post-natal Depression Scale (EPDS). In the second stage, mothers with EPDS scores > or =9 and a randomized sample of 16% with EPDS <9 were explored through a structured clinical interview to diagnose DSM-IV PND (major and minor depression). Variables were entered into stepwise regression models. A total of 1,201 women were recruited and did the EPDS; 261 women with EPDS scores > or =9 and 151 with EPDS scores <9 were selected. Three hundred and thirty-four women agreed to be interviewed and 100 were diagnosed with PND. Family caregiver role (defined as women who have to take care of handicapped or ill relatives) was associated with a 4.4-fold increase in risk for major PND (OR: 4.39, 95%CI: 1.10 17.38). Premenstrual syndrome was identified as an independent risk factor for major and minor PND (OR: 1.81, 95%CI: 1.03-3.18). Moreover, previous depression, poor partner relationship, and lower social support were also confirmed as risk factors for PND. Both family caregiver role and premenstrual syndrome should be considered for inclusion in the rating scales of pregnant women at risk for PND. PMID- 18506576 TI - Conserved network properties of helical membrane protein structures and its implication for improving membrane protein homology modeling at the twilight zone. AB - Homology modeling techniques remain an important tool for membrane protein studies and membrane protein-targeted drug development. Due to the paucity of available structure data, an imminent challenge in this field is to develop novel computational methods to help improve the quality of the homology models constructed using template proteins with low sequence identity. In this work, we attempted to address this challenge using the network approach developed in our group. First, a structure pair dataset of 27 high-resolution and low sequence identity (7-36%) comparative TM proteins was compiled by analyzing available X ray structures of helical membrane proteins. Structure deviation between these pairs was subsequently confirmed by calculating their backbone RMSD and comparing their potential energy per residue. Next, this dataset was further studied using the network approach. Results of these analyses indicated that the network measure applied represents a conserved feature of TM domains of similar folds with various sequence identities. Further comparison of this salient feature between high-resolution template structures and their homology models at the twilight zone suggested a useful method to utilize this property for homology model refinement. These findings should be of help for improving the quality of homology models based on templates with low sequence identity, thus broadening the application of homology modeling techniques in TM protein studies. PMID- 18506577 TI - 2D and 3D QSAR studies of diarylpyrimidine HIV-1 reverse transcriptase inhibitors. AB - 2D and 3D QSAR studies were applied on a set of 28 diarylpyrimidine derivatives to model and understand their HIV-1 reverse transcriptase (RT) inhibitory activities. Special cares were taken to build our set of molecules according to their bioactive conformations which is crucial to elaborate good QSAR models. 2D QSAR was performed using the heuristic method in CODESSA which had led to a linear model (R (2) = 0.928 and s (2) = 0.015) between the inhibitory activity and five descriptors. CoMFA and CoMSIA models were established using SYBYL package of programs. The better predictive ability of the CoMSIA model (q (2) = 0.730) over the CoMFA model (q (2) = 0.597) was assigned to the large contribution of hydrogen-bonding interactions to the inhibitory activity. CoMSIA physicochemical properties are in agreement with the 2D QSAR descriptors. The CoMSIA PLS contour surfaces were mapped to the binding pocket of the RT and showed that the results obtained by the 2D and 3D models are in respect with the protein environment. This link permitted us to validate our model and give important insights for the structure activity interpretations. These results will guide further structural modification and prediction of new HIV-1 RT inhibitors. PMID- 18506578 TI - Improved residual water suppression: WET180. AB - Water suppression in biological NMR is frequently made inefficient by the presence of faraway water that is located near the edges of the RF coil and experiences significantly reduced RF field. WET180 (WET with 180 degrees pulse toggling) is proposed to cancel the faraway water contribution to the residual solvent signal. The pulse sequence incorporates a modification of the last WET selective pulse to accommodate insertion of a toggled 180 degrees inversion pulse so that the original WET selective pulse angles are effectively preserved. Compared with existing WET methods, WET180 has the advantages of easy implementation, improved residual water suppression, clean spectral phase properties, and good signal intensity retention. WET180 is expected to be most useful in observing resonances close to water in samples containing biological molecules. In addition, the principle of WET180 can be applied in multidimensional experiments to improve residual water suppression and reduce artifacts around water. PMID- 18506579 TI - Double quantum filtering homonuclear MAS NMR correlation spectra: a tool for membrane protein studies. AB - 13C homonuclear correlation spectra based on proton driven spin diffusion (PDSD) are becoming increasingly important for obtaining distance constraints from multiply labeled biomolecules by MAS NMR. One particular challenging situation arises when such constraints are to be obtained from spectra with a large natural abundance signal background which causes detrimental diagonal peak intensities. They obscure cross peaks, and furthermore impede the calculation of a buildup rates matrix which may be used to derive distance constraints, as carried out in "NMR crystallography". Here, we combine double quantum (DQ) filtering with 13C 13C dipolar assisted rotational resonance (DARR) experiments to yield correlation spectra free of natural abundance contributions. Two experimental schemes, using DQ filtering prior to evolution (DOPE), and after mixing (DOAM), have been evaluated. Diagonal peak intensities along the spectrum diagonal are removed completely, and crosspeaks close to the diagonal are easily identifiable. For DOAM spectra with negligible mixing times, it is possible to carry out 'assignment walks' which simplify peak identification substantially. The method is demonstrated on 13C-cys labeled proteorhodopsin, a 27 kDa membrane protein. The magnetization transfer characteristics were studied using buildup curves obtained on uniformly 13C labelled crystalline tripeptide MLF. Our data show that DQ filtered DARR experiments pave the way for obtaining through space constraints for structural studies on ligands, bound to membrane receptors, or on small fragments within large proteins. PMID- 18506580 TI - The Arabidopsis thaliana carboxyl-terminal domain phosphatase-like 2 regulates plant growth, stress and auxin responses. AB - More than 20 genes in the Arabidopsis genome encode proteins similar to phosphatases that act on the carboxyl-terminal domain (CTD) of RNA polymerase II. One of these CTD-phosphatase-like (CPL) proteins, CPL2, dephosphorylates CTD-Ser5 PO4 in an intact RNA polymerase II complex and contains a double-stranded (ds) RNA-binding motif (DRM). Although the dsRNA-binding activity of CPL2 DRM has not been shown to date, T-DNA insertion mutants that express CPL2 variants lacking either a part of DRM (cpl2-1) or the entire DRM (cpl2-2) exhibited leaf expansion defects, early flowering, low fertility, and increased salt sensitivity. cpl2 mutant plants produced shorter hypocotyls than wild-type plants in the light, but were indistinguishable from wild type in the dark. CPL2 was expressed in shoot and root meristems and vasculatures, expanding rosette leaves, and floral organs suggesting a focal role for growth. Microarray and RT-PCR analyses revealed that basal levels of several auxin-responsive transcripts were reduced in cpl2. On the other hand, the levels of endogenous auxin and its conjugates were similar in wild type and cpl2. Overexpression of ARF5 but not all activator ARF transcription factors restored the auxin-responsive DR5-GUS reporter gene expression and the leaf expansion of cpl2 mutant plants but not early flowering phenotype. These results establish CPL2 as a multifunctional regulator that modulates plant growth, stress, and auxin responses. PMID- 18506581 TI - Functional analysis of cotton orthologs of GA signal transduction factors GID1 and SLR1. AB - Gibberellic acid (GA) is both necessary and sufficient to promote fiber elongation in cultured fertilized ovules of the upland cotton variety Coker 312. This is likely due to the temporal and spatial regulation of GA biosynthesis, perception, and subsequent signal transduction that leads to alterations in gene expression and morphology. Our results indicate that the initiation of fiber elongation by the application of GA to cultured ovules corresponds with increased expression of genes that encode xyloglucan endotransglycosylase/hydrolase (XTH) and expansin (EXP) that are involved in promoting cell elongation. To gain a better understanding of the GA signaling components in cotton, that lead to such changes in gene expression, two GA receptor genes (GhGID1a and GhGID1b) and two DELLA protein genes (GhSLR1a and GhSLR1b) that are orthologous to the rice GA receptor (GID1) and the rice DELLA gene (SLR1), respectively, were characterized. Similar to the GA biosynthetic genes, expression of GhGID1a and GhGID1b is under the negative regulation by GA while GA positively regulates GhSLR1a. Recombinant GST-GhGID1s showed GA-binding activity in vitro that was augmented in the presence of GhSLR1a, GhSLR1b, or rice SLR1, indicating complex formation between the receptors and repressor proteins. This was further supported by the GA dependent interaction of these proteins in yeast cells. Ectopic expression of the GhGID1a in the rice gid1-3 mutant plants rescued the GA-insensitive dwarf phenotype, which demonstrates that it is a functional GA receptor. Furthermore, ectopic expression of GhSLR1b in wild type Arabidopsis led to reduced growth and upregulated expression of DELLA-responsive genes. PMID- 18506582 TI - An optimal DNA pooling strategy for progressive fine mapping. AB - We present a cost-effective DNA pooling strategy for fine mapping of a single Mendelian gene in controlled crosses. The theoretical argument suggests that it is potentially possible for a single-stage pooling approach to reduce the overall experimental expense considerably by balancing costs for genotyping and sample collection. Further, the genotyping burden can be reduced through multi-stage pooling. Numerical results are provided for practical guidelines. For example, the genotyping effort can be reduced to only a small fraction of that needed for individual genotyping at a small loss of estimation accuracy or at a cost of increasing sample sizes slightly when recombination rates are 0.5% or less. An optimal two-stage pooling scheme can reduce the amount of genotyping to 19.5%, 14.5% and 6.4% of individual genotyping efforts for identifying a gene within 1, 0.5, and 0.1 cM, respectively. Finally, we use a genetic data set for mapping the rice xl(t) gene to demonstrate the feasibility and efficiency of the DNA pooling strategy. Taken together, the results demonstrate that this DNA pooling strategy can greatly reduce the genotyping burden and the overall cost in fine mapping experiments. PMID- 18506583 TI - Impact of two treatments of a formulation of Beauveria bassiana (Deuteromycota: Hyphomycetes) conidia on Varroa mites (Acari: Varroidae) and on honeybee (Hymenoptera: Apidae) colony health. AB - Bee colonies in southern France were treated with conidia (asexual spores) from two strains of Beauveria bassiana, an entomopathogenic fungus. One strain was commercial (GHA) and the other had been isolated from Varroa mites in the region (Bb05002). Objectives were to evaluate treatment effect on colony weight, adult bee mass, capped brood, and on Varroa fall onto sticky boards. Treatments included conidia formulated with either carnauba or candelilla wax powder, candelilla wax powder alone, or control; in two treatment groups formulation was applied a second time after one week. Treatment did not affect colony health. Colonies treated twice with Bb05002 conidia and carnauba wax powder had significantly higher mite fall compared to colonies treated with blank candelilla wax powder. The proportion of fallen mites that were infected in both conidia treatments was higher than controls for 18 days after the second treatment. The number of fungal propagules on the bees themselves remained elevated for about 14 days after the second treatment. These results were compared to published results from previous experiments with regard to infection duration. PMID- 18506584 TI - Using RNA interference to determine the role of varisin in the innate immune system of the hard tick Dermacentor variabilis (Acari: Ixodidae). AB - Defensins are an important component of the innate immune system of ticks. These small peptides are produced by various genera of ticks, and expressed in various tissues. In this study we used RNA interference to silence the expression of the defensin varisin produced by the hemocytes of the American dog tick, Dermacentor variabilis. Ticks were injected with double stranded varisin RNA prior to being placed on a rabbit. After feeding, the ticks were removed, bled, and the hemolymph plasma and hemocytes separated. Hemocytes were screened for the presence (or absence) of both varisin transcript and peptide. Varisin peptide was below detectable levels and the transcript showed a greater than 99% knockdown. The antimicrobial activity of the hemolymph plasma was reduced 2-4 fold compared to that of control injected ticks indicating varisin accounts for a large portion of the antimicrobial activity of the hemolymph. PMID- 18506585 TI - Pulmonary lymphangitic sarcomatosis and a review of the literature. AB - Intrapulmonary spread of a sarcoma via lymphatics is a rare cause of death in a young adult. A 31-year old man was admitted to our hospital complaining of dyspnea and malaise of 2 months' duration. A chest radiography revealed bilateral hilar enlargement, and reticulonodular infiltrations. Thoracic CT-scans demonstrated mediastinal lymphadenopathy, thickening of interlobular septa, polygonal lines, and thickening of bronchovascular bundles. The diagnosis was made by open-lung biopsy. The patient died within 3 months after diagnosis. Pulmonary lymphangitic sarcomatosis is a rare but important manifestation of an angiosarcoma. Optimal treatment of these patients is not well defined, but a trial of chemotherapy may be warranted. PMID- 18506586 TI - Histone deacetylase inhibitors: the anticancer, antimetastatic and antiangiogenic activities of AN-7 are superior to those of the clinically tested AN-9 (Pivanex). AB - Histone deacetylase inhibitory prodrugs that are metabolized to butyric acid and formaldehyde possess antineoplastic properties and low toxicity. We sought to characterize the antiangiogenic and antimetastatic activities of two lead prodrugs, pivaloyloxymethyl butyrate (AN-9) and butyroyloxymethyl-diethyl phosphate (AN-7) in murine cancer models. In the sc implanted human colon carcinoma HT-29 xenograft model AN-7, exhibited superior anticancer activity compared to AN-9, as was evident by the significantly greater inhibition of tumor growth and reduction of serum CEA. AN-7 was also more effective in reducing mean vessel density (MVD) by 7-fold, bFGF, Ki-67 (7-fold) and HIF-1alpha in immunohistochemically stained tumor sections. Semi-quantitative evaluation of the levels of bFGF, HDAC1 and HIF-1alpha by Western blot analysis showed a decrease in expression only in the tumors of mice treated with AN-7. The level of bFGF was reduced 3-fold in the tumor and that of TIMP1 was elevated (by 3-fold) in the serum of AN-7 treated mice. In a 4T1 metastatic breast carcinoma model, AN-7 inhibited the formation of lung lesions by 76% and AN-9 by 47%, further demonstrating the greater efficacy of AN-7 compared to AN-9 (P<0.02). Both AN-7 and AN-9 exhibited antimetastatic and antiangiogenic activities by reducing vascularization, bFGF expression and HIF-1alpha. Yet, AN-7 was more potent than AN-9. PMID- 18506587 TI - Intraosseous injection of RM1 murine prostate cancer cells promotes rapid osteolysis and periosteal bone deposition. AB - The molecular mechanisms associated with prostate cancer (PCa) progression within bone remain a topic of intense investigation. With the availability of transgenic mouse strains, a model of PCa for use in immune competent/transgenic mice would be highly beneficial. This study was designed to explore the utility of RM1 mouse PCa cells in investigations of tumor:bone interactions. The efficacies of several implantation techniques were examined for reliably producing intra-bone RM1 tumor growth and bone lesion formation in immune competent mice. Longitudinal monitoring of bone remodeling and lesion phenotypes was conducted by microcomputed tomography (muCT) and histological analyses. Our results indicate that direct intrabone injections of RM1 cells are necessary for tumor growth within bone and direct implantation promotes the rapid development of osteolytic bone lesions with periosteal bone deposition post-cortical breach. In vitro, RM1 cells promote the proliferation of osteoblast (MC3T3-E1) and osteoclast (Raw264.7) progenitors in a dose dependent manner. Conditioned culture media from RM1 cells appears to promote earlier expression of genes/proteins associated with osteoblastic differentiation. While clearly stimulating osteoclast function in vivo, RM1 cells had little effect on differentiation and tartate resistant acid phosphatase (TRAP) expression by Raw264.7 cells. These data, coupled with in vivo muCT images, indicate the ability of RM1 cells to induce mixed, yet predominentally osteolytic, responses in bone and illustrate the potential of RM1 cells as a model of investigating prostate tumor:stroma interactions in immune competent/transgenic mice on a C57BL/6 background. PMID- 18506589 TI - A flow-through capnometer for obstructive sleep apnea. AB - INTRODUCTION: Capnogram is often distorted due to aspiration of expired gas when a sidestream capnometer is used for non-intubated, spontaneously breathing condition. The purpose of this study was to make a flow-through capnometer without aspiration and to check if this capnometer precisely detected apnea during obstructive sleep apnea (OSA). METHODS: (1) Flow-through capnometer The capnometer consisted of a flow-through etCO(2) sensor, cap-ONE, and an accompanying capnometer. The size of cap-ONE was small enough to be fitted under the nose where gas expired from the nose and the mouth passed through. Thus, the expired gas to be measured is directly blown into the cap-ONE. (2) The cap-ONE using a spontaneously breathing model Capnograms obtained by the cap-ONE and sidestream capnometers during nasal and oral breathing under normal and reduced ventilation were compared with a reference capnogram. (3) Clinical study with OSA patients With nineteen OSA patients capnograms during apnea events diagnosed as OSA by polysomnography were examined using the cap-ONE. (4) Simulation study with an OSA model Apnea in which inspiratory flow was zero and small expiratory flows repeated was produced. Capnograms and apnea detection were compared between the cap-ONE and sidestream capnometers. RESULTS: In the spontaneouly breathing model capnograms and etCO(2) of the cap-ONE during nasal and oral breathing were almost identical with the reference capnogram but those of sidetream capnometers during oral breathing were significantly reduced. In the clinical study 41% of total OSA events showed capnograms with prolonged and elevated phase with small ripples. In a simulation study reduction of CO(2) tension during no-inspiration was small and apnea was successfully detected with the cap-ONE. However, with sidestream capnometers the reduction of CO(2) tension was large and apnea was not detected. CONCLUSIONS: We concluded that the cap-ONE can record capnograms with minimum distortion and detect apnea reliably during OSA. PMID- 18506590 TI - Cell cycle-dependent regulation of telomere tethering in the nucleus. AB - It is well established that telomeres are tethered in the eukaryotic nucleus, but a detailed analysis of the regulation of telomere attachment throughout the cell cycle is still lacking. We show here that the telomeres in the macronucleus of the ciliate Stylonychia lemnae are bound to a sub-nuclear structure by an interaction of the telomere end-binding protein TEBPalpha with three SNS proteins that are integral parts of this structure. In the course of replication, the interaction of TEBPalpha with the SNS proteins is resolved and this process is regulated by cell cycle-specific phosphorylation of the SNS proteins. Our data can be incorporated into a mechanistic model for the regulation of telomere conformation and localization throughout the cell cycle. PMID- 18506591 TI - Distribution of randomly generated activity class characteristic substructures in diverse active and database compounds. AB - Substructures are among the most preferred molecular descriptors in chemoinformatics and medicinal chemistry. Conventional substructure-type descriptors are typically the result of well-defined design strategies. Previously, we have introduced Activity Class Characteristic Substructures (ACCS) derived from randomly generated molecular fragment populations and described their utility in similarity searching. Short ACCS fingerprints were found to perform surprisingly well on many compound classes when compared to more complex state-of-the-art 2D fingerprints. In order to elucidate potential reasons for the high predictive utility of ACCS, we have carried out a thorough analysis of their distribution in nine activity classes and nearly four million database compounds. We show that the discriminatory power of ACCS results from the rare occurrence of ACCS combinations in screening databases. PMID- 18506592 TI - Pharmacophore mapping of flavone derivatives for aromatase inhibition. AB - Aromatase, which catalyses the final step in the steroidogenesis pathway of estrogen, has been target for the design of inhibitor in the treatment of hormone dependent breast cancer for postmenopausal women. The extensive SAR studies performed in the last 30 years to search for potent, selective and less toxic compounds, have led to the development of second and third generation of non steroidal aromatase inhibitors (AI). Besides the development of synthetic compounds, several naturally occurring and synthetic flavonoids, which are ubiquitous natural phenolic compounds and mediate the host of biological activities, are found to demonstrate inhibitory effects on aromatase. The present study explores the pharmacophores, i.e., the structural requirements of flavones (Fig. 1) for inhibition of aromatase activity, using quantitative structure activity relationship (QSAR) and space modeling approaches. The classical QSAR studies generate the model (R (2) = 0.924, Q (2) = 0.895, s = 0.233) that shows the importance of aromatic rings A and C, along with substitutional requirements in meta and para positions of ring C for the activity. 3D QSAR of Comparative Molecular Field Analysis (CoMFA, R (2) = 0.996, R(2)(cv) = 0.791) and Comparative Molecular Similarity Analysis (CoMSIA, R (2) = 0.992, R(2)(cv) = 0.806) studies show contour maps of steric and hydrophobic properties and contribution of acceptor and donor of the molecule, suggesting the presence of steric hindrance due to ring C and R''-substituent, bulky hydrophobic substitution in ring A, along with acceptors at positions 11, and alpha and gamma of imidazole ring, and donor in ring C favor the inhibitory activity. Further space modeling (CATALYST) study (R = 0.941, Delta( cost ) = 96.96, rmsd = 0.876) adjudge the presence of hydrogen bond acceptor (keto functional group), hydrophobic (ring A) and aromatic rings (steric hindrance) along with critical distance among features are important for the inhibitory activity. PMID- 18506593 TI - Using AdaBoost for the prediction of subcellular location of prokaryotic and eukaryotic proteins. AB - In this paper, AdaBoost algorithm, a popular and effective prediction method, is applied to predict the subcellular locations of Prokaryotic and Eukaryotic Proteins-a dataset derived from SWISSPROT 33.0. Its prediction ability was evaluated by re-substitution test, Leave-One-Out Cross validation (LOOCV) and jackknife test. By comparing its results with some most popular predictors such as Discriminant Function, neural networks, and SVM, we demonstrated that the AdaBoost predictor outperformed these predictors. As a result, we arrive at the conclusion that AdaBoost algorithm could be employed as a robust method to predict subcellular location. An online web server for predicting subcellular location of prokaryotic and eukaryotic proteins is available at http://chemdata.shu.edu.cn/subcell/ . PMID- 18506594 TI - Increased protein carbonylation in leaves of Arabidopsis and soybean in response to elevated [CO2]. AB - While exposure of C3 plants to elevated [CO2] would be expected to reduce production of reactive oxygen species (ROS) in leaves because of reduced photorespiratory metabolism, results obtained in the present study suggest that exposure of plants to elevated [CO2] can result in increased oxidative stress. First, in Arabidopsis and soybean, leaf protein carbonylation, a marker of oxidative stress, was often increased when plants were exposed to elevated [CO2]. In soybean, increased carbonyl content was often associated with loss of leaf chlorophyll and reduced enhancement of leaf photosynthetic rate (Pn) by elevated [CO2]. Second, two-dimensional (2-DE) difference gel electrophoresis (DIGE) analysis of proteins extracted from leaves of soybean plants grown at elevated [CO2] or [O3] revealed that both treatments altered the abundance of a similar subset of proteins, consistent with the idea that both conditions may involve an oxidative stress. The 2-DE analysis of leaf proteins was facilitated by a novel and simple procedure to remove ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from soluble soybean leaf extracts. Collectively, these findings add a new dimension to our understanding of global change biology and raise the possibility that oxidative signals can be an unexpected component of plant response to elevated [CO2]. PMID- 18506595 TI - Magnetic resonance imaging: the preferred imaging method for evaluation of the right ventricle. PMID- 18506596 TI - Cloning and characterization of cDNA probes for the analysis of metallothionein gene expression in the Mediterranean bivalves: Ruditapes decussatus and Cerastoderma glaucum. AB - cDNA probes have been developed for subsequent use in monitoring the cadmium exposure of the clam Ruditapes decussatus and the cockle Cerastoderma glaucum using metallothionein (MT) gene expression in different tissues of these species. Two partial MT cDNAs were isolated from Ruditapes decussatus and Cerastoderma glaucum. The identification of the nucleotide sequences showed that the cDNAs consist of 480 bp coding 72 amino acid proteins containing 21 cysteine residues organized in Cys-X-Cys motifs as classically described for MTs. The induction of MT gene expression in CdCl(2) treated bivalves was confirmed by dot blot analysis and suggests a potential specific tissue expression rate. PMID- 18506597 TI - Lack of association between interleukin-13 gene polymorphisms (-1055 C/T and +2044 G/A) in Iranian patients with lung cancer. AB - Lung cancer is one of the leading causes of death from cancer. Both immune cells and tumor cells play a key role in lung cancer immunity by secretion of cytokines and developing type-2 cell-mediated immune response. IL-13 is an immunoregulatory cytokine affecting tumor immunosurveillance by deviation of immune response from Th1 to Th2. In the present study we sought to determine the association of single nucleotide polymorphisms (SNPs) of IL-13 gene at positions +2044 (G/A) and -1055 (C/T) and lung cancer. One hundred forty one patients and 113 controls were recruited; control group was subdivided into smoker and nonsmoker individuals for serum detection. Genotyping was carried out by PCR-RFLP assay and IL-13 detection by ELISA method. No statistically significant difference was found in the frequency of genotypes, alleles, and haplotypes at positions +2044 (G/A) and 1055 (C/T) of IL-13 gene between lung cancer patients and controls. Serum level of IL-13 was not detectable in both groups. The results of this study reveal that although +2044 (G/A) and -1055 (C/T) SNPs in IL-13 are implicated in some pulmonary processes, they do not confer susceptibility to lung cancer in Iranian population. PMID- 18506599 TI - Garlic allyl derivatives interact with membrane lipids to modify the membrane fluidity. AB - As a novel approach to the mode of medicinal action of garlic, its constituents were comparatively studied with respect to their interactions with membrane lipids to modify the membrane fluidity. Allyl derivatives rigidified tumor cell and platelet model membranes consisting of unsaturated phospholipids and cholesterol at 20-500 muM with the potency being diallyl trisulfide (DATS) > diallyl disulfide (DADS) by preferentially acting on the hydrocarbon cores of lipid bilayers. They were also effective in rigidifying candida cell model membranes prepared with ergosterol and phospholipids at 100-500 microM with the potency being DADS > DATS > diallyl sulfide (DAS), but not bacteria cell model membranes without ergosterol. Alliin, a precursor of these DASs, was not active on any membranes at 500 microM. Both relative intensity and selectivity in membrane effects correlated with those in antiproliferative, antiplatelet and antimicrobial effects. In cell culture experiments, membrane-active DASs inhibited the growth of tumor cells cultured for 24 and 48 h at 20-500 muM to show the potency being DATS > DADS, together with rigidifying cell membranes by acting on their deeper regions more intensively. However, membrane-inactive allyl derivatives were not growth-inhibitory on tumor cells. The membrane lipid interactions of DASs appear to be one of possible mechanisms underlying different effects of garlic. PMID- 18506598 TI - Severity of cardiovascular disease and health-related quality of life in men with prostate cancer: a longitudinal analysis from CaPSURE. AB - OBJECTIVE: To evaluate the influence of comorbid cardiovascular disease severity on health-related quality of life (HRQL) in men treated with radical prostatectomy (RP) or radiotherapy (RT) for early stage prostate cancer. METHODS: Subjects (n=830) with non-metastatic disease who had been diagnosed in 2000-2002 were drawn from Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE). We evaluated the influence of cardiovascular disease (CVD) severity on generic and disease-specific HRQL before and 6, 12, 18, and 24 months after treatment with RP or RT. HRQL was measured with the SF-36 and the UCLA Prostate Cancer Index. RESULTS: Men with moderate (n=193) or severe (n=51) cardiovascular disease had worse pre-treatment HRQL than did men without CVD (n=293) (P<0.01); HRQL scores were worse in men referred for RT. During 24 months of follow-up, men with moderate or severe CVD had worse SF-36 physical and mental component summaries and worse bowel function at all time points (P<0.05). Men with severe CVD also experienced a slower recovery in physical function (P=0.03) and sexual functioning (P=0.02) than did men without CVD. CONCLUSIONS: Prostate cancer patients with moderate to severe CVD have worse HRQL during follow-up. Those with severe CVD recover their physical and sexual functioning more slowly after treatment. PMID- 18506600 TI - CCN3: A novel function in vivo. PMID- 18506601 TI - Hsp60D is essential for caspase-mediated induced apoptosis in Drosophila melanogaster. AB - Apart from their roles as chaperones, heat shock proteins are involved in other vital activities including apoptosis with mammalian Hsp60 being ascribed proapoptotic as well as antiapoptotic roles. Using conditional RNAi or overexpression of Hsp60D, a member of the Hsp60 family in Drosophila melanogaster, we show that the downregulation of this protein blocks caspase dependent induced apoptosis. GMR-Gal4-driven RNAi for Hsp60D in developing eyes dominantly suppressed cell death caused by expression of Reaper, Hid, or Grim (RHG), the key activators of canonical cell death pathway. Likewise, Hsp60D-RNAi rescued cell death induced by GMR-Gal4-directed expression of full-length and activated DRONC. Overexpression of Hsp60D enhanced cell death induced either by directed expression of RHG or DRONC. However, the downregulation of Hsp60D failed to suppress apoptosis caused by unguarded caspases in DIAP1-RNAi flies. Furthermore, in DIAP1-RNAi background, Hsp60D-RNAi also failed to inhibit apoptosis induced by RHG expression. The Hsp60 and DIAP1 show diffuse and distinct granular overlapping distributions in the photoreceptor cells with the bulk of both proteins being outside the mitochondria. Depletion of either of these proteins disrupts the granular distribution of the other. We suggest that in the absence of Hsp60D, DIAP1 is unable to dissociate from effecter and executioner caspases, which thus remain inactive. PMID- 18506602 TI - Parameters for safer gambling behavior: examining the empirical research. AB - There have been claims that new gambling technology is hazardous to player health, and that technological interventions can alleviate gambling-related harm. In this paper, we systematically review the empirical research about the nexus between gambling and technology to evaluate the veracity of these claims. We use a public health perspective (i.e., the Epidemiologic Triangle) to organize and present study results (i.e., agent, host, and environment). This review intends to offer insight about emerging technology and identify areas that indicate a need for additional research. Forty-seven studies met our inclusion and exclusion criteria; a review of this body of work shows that attempts to develop and implement safety features for new gambling technology are promising, but methodologically are rudimentary and limited in scope. Increased attention to the dynamic interaction among host, agent, and environment factors hold potential to advance the field. In addition, improved study methods (e.g., longitudinal analyses of actual betting behavior), and collaboration among policymakers, manufacturers, and researchers can increase understanding of how new gambling technology affects the public health and stimulate new strategies for implementing effective public health interventions. PMID- 18506603 TI - Thioflavin T displays enhanced fluorescence selectively inside anionic micelles and mammalian cells. AB - Thioflavin T (ThT) has been widely employed to detect amyloid fibrils in tissues and recently in presence of SDS micelles. However, the contribution of membranes or micelles to ThT fluorescence has never been investigated. In this paper, we show for the first time that the anionic micellar microenvironment of SDS has a profound impact on the absorption and fluorescence spectra of ThT in sharp contrast to cationic (CTAB) and neutral micelles (Triton X-100 & Tween 20). Unlike CTAB or Triton X-100 or Tween 20 micelles, formation of SDS micelles shifts the lambdamax for ThT absorption from 412 nm in buffer to 428 nm inside the micelle, with a 28% increase in the peak molar absorptivity and an approximately 13 fold increase in ThT fluorescence (lambdamax=489 nm). Extending these observations to cell plasma membranes, we show that ThT can quickly enter and appear selectively fluorescent inside mammalian cells like BHK21 and HT29, against a dark background owing to negligible fluorescence from free ThT in aqueous medium. The above results suggest that ThT can be a useful probe for live cell imaging and for selectively labeling micelles on the basis of the charge in the polar headgroup. PMID- 18506604 TI - Examining the incidence of and time to recidivism within the risk contingency framework: a 20-year follow up study. AB - This study examined the extent to which the contingencies risk framework (consisting of dispositional, historical, contextual, and clinical domains) predicted detected recidivism (i.e., arrest or conviction). Secondary data were extracted on 413 prisoners who underwent a psychiatric evaluation, were assessed on the risk domains, and followed up over 20 years. There were 273 (66.1%) cases of recidivism for serious offenses (e.g., assaults). Criminal career outcomes examined included: years to and the incidence of recidivism. Statistics showed that chance classification of the incidence of recidivism was 21% more accurate for the recursive partitioning than the bilinear model. These results are consistent with the contingencies risk framework, support its use over linear models, and highlight its predictive utility. PMID- 18506606 TI - Effect of presenting rhythm on image integration to direct catheter ablation of atrial fibrillation. AB - INTRODUCTION: Magnetic resonance (MR) imaging of the left atrium (LA) can be integrated with electroanatomic mapping systems to guide catheter ablation of atrial fibrillation (AF). The usefulness of this technique is dependent on the accuracy of image integration. OBJECTIVE: The aim of this study is to determine the effect of heart rhythm at the time of pre-procedure MR imaging and heart rhythm at the time of ablation on integration error. METHODS: Fifty-two consecutive patients who underwent catheter ablation for AF were included. All patients underwent MR imaging of LA and pulmonary veins and image integration with real-time electroanatomic mapping. The rhythm at the time of MR imaging and on the day of ablation was recorded. CARTO-Merge software (Biosense-Webster) was used to calculate the average accuracy of integration of electroanatomic points with MR-derived reconstructions. RESULTS: There was no significant difference in integration error between patients who were in AF at the time of their MR vs. those who were in sinus rhythm at the time of their MR (1.76 +/- 0.26 vs. 1.88 +/ 0.31 mm, p = 0.15). There was also no significant difference in integration error between patients who were in concordant vs. discordant rhythms at the time of MR vs. day of ablation (1.81 +/- 0.23 vs. 1.89 +/- 0.32 mm, p = 0.40). There was a trend toward less integration error between patients who were in AF on the day of ablation vs. those in sinus rhythm (1.74 +/- 0.26 vs. 1.89 +/- 0.31 mm, p = 0.07). CONCLUSIONS: Image integration can be performed to direct catheter ablation of AF regardless of the rhythm at the time of imaging and ablation. PMID- 18506607 TI - Interatrial septal puncture without the use of fluoroscopy-reducing ionizing radiation in left atrial ablation procedures. AB - Ablation for atrial fibrillation is now widespread and may involve the delivery of relatively high doses of ionizing radiation to both patients and operators. Until now, atrial septal puncture has mandated the use of either fluoroscopy or additional expensive imaging modalities for its safe deployment. We describe a novel technique using the EnSite NavX three-dimensional imaging system for visualization of the transseptal needle throughout the puncture-without the need for either fluoroscopy or additional imaging. This may be important in reducing the need for fluoroscopy for left atrial ablation procedures. PMID- 18506608 TI - Pulmonary vein calcification by EBCT in patients with drug refractory nonvalular atrial fibrillation. AB - INTRODUCTION: Pulmonary veins in patients with atrial fibrillation (AF) have been shown to be highly arrhythmogenic. Calcification in these veins may play an adjunctive role in the pathogenesis of AF. METHODS AND RESULTS: A case control study was performed in patients with drug refractory nonvalvular AF whose preablation computed tomography chest scans demonstrated pulmonary vein (PV) calcification. Eight out of 48 patients with PV calcification were compared to 50 patients without AF who underwent electron beam computed tomography coronary artery calcium scores. These patients were matched for age, gender, coronary artery calcium scores, and the presence of PV calcification. The mean age of the combined group was 57 +/- 9 years and 60% were men. The mean total PV calcium score was significantly higher at 199 +/- 112 in patients with AF compared to 106 +/- 52 in controls (p = 0.018). Men had significantly higher total PV calcium score than women in both groups. CONCLUSION: Total PV calcium score was significantly higher in patients with atrial fibrillation. Increased PV calcification may play an adjunctive role in the pathogenesis in initiating and maintaining AF. PMID- 18506609 TI - Parametric and non-parametric modeling of short-term synaptic plasticity. Part I: Computational study. AB - Parametric and non-parametric modeling methods are combined to study the short term plasticity (STP) of synapses in the central nervous system (CNS). The nonlinear dynamics of STP are modeled by means: (1) previously proposed parametric models based on mechanistic hypotheses and/or specific dynamical processes, and (2) non-parametric models (in the form of Volterra kernels) that transforms the presynaptic signals into postsynaptic signals. In order to synergistically use the two approaches, we estimate the Volterra kernels of the parametric models of STP for four types of synapses using synthetic broadband input-output data. Results show that the non-parametric models accurately and efficiently replicate the input-output transformations of the parametric models. Volterra kernels provide a general and quantitative representation of the STP. PMID- 18506610 TI - Simulation system of spinal cord motor nuclei and associated nerves and muscles, in a Web-based architecture. AB - A Web-based simulation system of the spinal cord circuitry responsible for muscle control is described. The simulator employs two-compartment motoneuron models for S, FR and FF types, with synaptic inputs acting through conductance variations. Four motoneuron pools with their associated interneurons are represented in the simulator, with the possibility of inclusion of more than 2,000 neurons and 2,000,000 synapses. Each motoneuron action potential is followed, after a conduction delay, by a motor unit potential and a motor unit twitch. The sums of all motor unit potentials and twitches result in the electromyogram (EMG), and the muscle force, respectively. Inputs to the motoneuron pool come from populations of interneurons (Ia reciprocal inhibitory interneurons, Ib interneurons, and Renshaw cells) and from stochastic point processes associated with descending tracts. To simulate human electrophysiological experiments, the simulator incorporates external nerve stimulation with orthodromic and antidromic propagation. This provides the mechanisms for reflex generation and activation of spinal neuronal circuits that modulate the activity of another motoneuron pool (e.g., by reciprocal inhibition). The generation of the H-reflex by the Ia motoneuron pool system and its modulation by spinal cord interneurons is included in the simulation system. Studies with the simulator may include the statistics of individual motoneuron or interneuron spike trains or the collective effect of a motor nucleus on the dynamics of muscle force control. Properties associated with motor-unit recruitment, motor-unit synchronization, recurrent inhibition and reciprocal inhibition may be investigated. PMID- 18506611 TI - Risk reduction for HIV-positive African American and Latino men with histories of childhood sexual abuse. AB - While the HIV epidemic has disproportionately affected African American and Latino men who have sex with men (MSM), few HIV prevention interventions have focused on African American and Latino men who have sex with both men and women (MSMW). Even fewer interventions target HIV-positive African American and Latino MSM and MSMW with histories of childhood sexual abuse (CSA), a population that may be vulnerable to high-risk sexual behaviors, having multiple sexual partners, and depression. The Men's Health Project, a small randomized clinical trial, compared the effects of two 6-session interventions, the Sexual Health Intervention for Men (S-HIM), guided by social learning theory and aimed at decreasing high-risk sexual behaviors, number of sexual partners, and depressive symptoms, and a standard health promotion control (SHP). A community sample of 137 HIV-positive gay and non-gay identifying African American and Latino MSM and MSMW with histories of CSA was recruited. Results were based on an "intent to treat" analyses of baseline to post, 3 and 6 month follow-ups. The sample as a whole reported reductions in sexual risk behaviors and number of sexual partners from baseline to post-test, and from the 3 to 6 month follow-ups, although the decrease in sexual risk behavior from baseline to post-test was significant only for S-HIM participants. No significant differences between conditions were reported for depressive symptoms, but the total sample reported a significant decrease at 6 months. These findings highlight the importance of addressing sexual decision-making and psychological adjustment for ethnic men, while being sensitive to CSA histories and sexual minority status, and suggest the need to develop additional strategies to heighten HIV risk reduction over time. PMID- 18506612 TI - Bisexually active Black men in the United States and HIV: acknowledging more than the "Down Low". AB - HIV is disproportionately impacting Black men who have sex with men and heterosexual women in the United States. Current speculation posits a "bisexual bridge" of HIV transmission connecting these two subpopulations of the Black community. Specifically, bisexually active Black men who identify as heterosexual but do not disclose their same-sex behavior, or "down low" (DL) men, have received the most attention and blame as the primary group fueling this epidemic. This essay explores the current knowledge and limitations of public health research on bisexually active Black men. Implications for future research initiatives are discussed. PMID- 18506613 TI - Psychosocial correlates of unprotected sex without disclosure of HIV-positivity among African-American, Latino, and White men who have sex with men and women. AB - African-American, Latino, and White men who have sex with men and women (MSMW) may be a bridge of HIV transmission from men to women. Very little research has directly compared culturally specific correlates of the likelihood of unprotected sex among MSMW. The present study examined psychosocial correlates of unprotected sex without disclosure of HIV status with male and female partners among 50 African American, 50 Latino, and 50 White HIV-positive MSMW recruited from AIDS service organizations in Los Angeles County. Multivariate logistic regressions were conducted to examine relationships of race/ethnicity and psychosocial variables (e.g., condom attitudes, self-efficacy for HIV disclosure, sexual identification) to unprotected sex without disclosure of HIV status, for male and female partners separately. For female partners, different effects emerged by race/ethnicity. Among African-Americans, less exclusively homosexual identification and low self-efficacy for disclosure of HIV status to female partners were associated with unprotected sex without disclosure; among Latinos, less exclusively homosexual identification and negative attitudes about condoms were significant. Participants who were more exclusively homosexually identified, who held less positive condom attitudes, and who had low self-efficacy for disclosure to female partners were more likely to have unprotected sex without disclosure of HIV status to male partners. Culturally tailored community-level interventions may help to raise awareness about HIV and bisexuality, and decrease HIV and sexual orientation stigma, thereby increasing African-American and Latino MSMW's comfort in communicating with their female partners about sexuality, HIV and condoms. Addressing norms for condom use and disclosure between male partners is recommended, especially for homosexually identified MSMW. PMID- 18506614 TI - "And then there was the Down Low": introduction to Black and Latino male bisexualities. AB - Although a recent proliferation of mass media has drawn attention to "the new Down Low phenomenon" (presumably "secretive" homosexuality among married Black men), relatively little research has explored bisexual behavior and identity among ethnic minority men in the United States or elsewhere. Although the study of bisexuality in Black and Latino men is significant in its own right, disproportionate rates of HIV/AIDS among these men make the current dearth of scientific information even more urgent and concerning. In this special section, we have compiled a diverse array of empirical and theoretical perspectives on Black and Latino male bisexualities. A wide range of information on the individual, social, and sexual lives of these men, and potential relations to risk behavior, are presented. This article introduces this new body of work and offers suggestions for future research directions for culturally appropriate interventions for Black and Latino bisexual men. PMID- 18506615 TI - The embodiment of tourism among bisexually-behaving Dominican male sex workers. AB - While theories of "structure" and social inequality have increasingly informed global health efforts for HIV prevention--with growing recognition of the linkages between large-scale political and economic factors in the distribution and impact of the HIV/AIDS epidemic--there is still little theorization of precisely how structural factors shape the very bodies and sexualities of specific populations and groups. In order to extend the theoretical understanding of these macro-micro linkages, this article examines how the growth of the tourism industry in the Dominican Republic has produced sexual practices and identities that reflect both the influence of large-scale structural processes and the resistant responses of local individuals. Drawing on social science theories of political economy, embodiment, and authenticity, I argue that an understanding of patterns of sexuality and HIV risk in the region requires analysis of how political-economic transformations related to tourism intersect with the individual experiences and practices of sexuality on the ground. The analysis draws on long-term ethnographic research with bisexually behaving male sex workers in two cities in the Dominican Republic, including participant observation, in-depth interviews, focus groups, and surveys. By examining the global and local values placed on these men's bodies and the ways sex workers use their bodies to broker tourists' pleasure, we may better understand how the large scale structures of the tourism industry are linked to the specific meanings and practices of sexuality. PMID- 18506616 TI - Sexual behaviors of non-gay identified non-disclosing men who have sex with men and women. AB - The sexual behaviors of non-gay identified men who have sex with men and women (MSMW) who do not disclose their same-sex behavior to their female partners (referred to by some as men "on the down low") were examined, including the potential for these men to serve as a "bisexual bridge" for HIV and STD acquisition and transmission. Self-reported sexual behavior data were collected as part of an exploratory study of an ethnically diverse (41% African American, 35% Latino, 22% White, and 2% Asian) sample of 46 non-gay identified, non disclosing MSMW in New York City. Men reported significantly more male than female sexual partners, but more frequent sex with females. The prevalence and frequency of unprotected sex did not differ significantly between male and female partners in the past 3 months. Men reported risk behavior more often with steady female and steady male partners (e.g., wives, girlfriends, boyfriends) than with more casual male and female partners (regular uncommitted partners, one-night stands). Men, especially African American/Black men, were significantly less likely to report receptive sexual behaviors with men than insertive behaviors. Unprotected sex was common with male and female partners in the past 3 months (e.g., 35% of anal sex with men; 50% of vaginal sex). Indeed, 22% of the sample (38% of those who had both recent males and female partners) reported both unprotected vaginal sex and unprotected anal sex with a male partner during the past 3 months. Although the study was limited by a small convenience sample, the findings suggest that non-gay identified non-disclosing MSMW are at risk for the acquisition and transmission of HIV and STDs, and may serve as a potential bridge, suggesting the need for further research and intervention targeting this unique population. PMID- 18506617 TI - Improving integrated general medical and mental health services in community based practices. AB - The historical fragmentation of physical and mental health services has impeded efforts to improve quality and outcomes of care for persons with mental disorders. However, there is little information on effective strategies that might reduce fragmentation and improve integrated services within non-academic, community-based healthcare settings. Twenty-three practices from across the U.S. participated in a learning community meeting designed to identify barriers to integrated care and strategies for reducing such barriers. Barriers were initially identified based on a quantitative survey of organizational factors. Focus groups were used to elaborate on barriers to integrated care and to identify strategies for reducing barriers that are feasible in community-based settings. Participants identified key barriers, including administrative (e.g., lack of common medical records for mental health and general medical conditions), financial (e.g., lack of reimbursement codes to bill for mental health and general medical care in the same setting), and clinical (e.g., lack of an integrated care protocol). Top strategies recommended by participants included templates (i.e., for memoranda of understanding) to allow providers to work across practice settings, increased medical record security to enable a common medical record between mental health and general medical care, working with state Medicaid agencies to establish integrated care reimbursement codes, and guidance in establishing workflows between different providers (i.e., avoid duplication of tasks). Strategies to overcome barriers to integrated care may require cooperation across different organizational levels, including administrators, providers, and health care payers in order for integrated care to be established and sustained over time. PMID- 18506618 TI - Evaluation of treatment programs for dual disorder individuals: modeling longitudinal and mediation effects. AB - This study evaluated the effectiveness of the three approaches for treating dual disorder clients who were homeless at intake: integrated assertive community treatment (IACT), assertive community treatment only (ACTO), and standard care (SC). Multilevel Random Coefficient Modeling (MRCM) was used to analyze longitudinal effects and to identify mediators of significant treatment effects. The outcome variables were consumer satisfaction, stable housing, psychiatric symptoms, and substance abuse. The eight mediators were service utilization variables: program contacts, phone contacts, substance abuse contacts, assistance with activities of daily living, transportation assistance, help finding permanent housing, help with emotional problems, and medication assistance. The 191 eligible participants were randomly assigned to one of the three conditions and followed for a period of 30 months. Both ACTO and IACT produced better outcomes than SC on consumer satisfaction and stable housing. There were no differences on any of the outcome variables between ACTO versus IACT when comparing main effects. However, there were several treatment by time interactions. In addition, there were many mediation effects. PMID- 18506619 TI - Cancer stem/progenitor cell active compound 8-quinolinol in combination with paclitaxel achieves an improved cure of breast cancer in the mouse model. AB - Increasing evidence suggests that breast cancer is caused by cancer stem cells and the cure of breast cancer requires eradication of breast cancer stem cells. In this study, we established and characterized a sphere culture model derived from side population cells from the human breast cancer cell line MCF7. The sphere culture could be maintained long term and was enriched in cells expressing known breast cancer stem cell marker CD44+CD24-. These sphere cells showed higher colony formation ability in vitro and higher tumorigenicity in vivo than MCF7 cells, suggesting the enrichment of breast cancer stem/progenitor cells. To identify compounds that preferentially inhibit the sphere cells, we performed a compound library screening. Two lead compounds, NSC24076 and NSC125034 and an analog of NSC125034, 8-quinolinol (8Q), were identified as having preferential activity against the sphere cells. 8Q showed some antitumor activity alone but had much better therapeutic effect and relapse prevention when combined with paclitaxel than either 8Q or paclitaxel alone in both MCF7 and MDA-MB-435 xenograft models. We propose that compounds selectively targeting cancer stem/progenitor cells when combined with standard chemotherapy drugs may produce an improved treatment of cancer without significant relapse. PMID- 18506620 TI - Analysis of chemotherapy-induced amenorrhea rates by three different anthracycline and taxane containing regimens for early breast cancer. AB - PURPOSE: Chemotherapy-induced amenorrhea (CIA) by newer taxane-containing regimens was evaluated in early breast cancer (EBC) patients. METHODS: A prospective cohort of 122 premenopausal EBC patients participated in a phase III trial of preoperative docetaxel/capecitabine (TX) versus doxorubicin/cyclophosphamide (AC); 34 patients received adjuvant AC followed by paclitaxel (T) and 129 patients received 5 fluorouracil/doxorubicin/cyclophosphamide (FAC). RESULTS: The CIA rate was 90.2% with TX/AC, 73.5% with AC followed by T, and 72.1% with FAC at 1 year (P = 0.002), and 66.7%, 73.3%, and 58.9%, respectively, at 3 years (P = 0.268). At one year, age (P < 0.001) and taxane use (P = 0.002), and after two years, age and tamoxifen use were significant factors for CIA in multivariate analysis. Serum estradiol and follicle-stimulating hormone levels were significantly correlated with menstrual status, age, and tamoxifen use. CONCLUSION: Taxanes resulted in higher CIA rates in the first year, but age and tamoxifen use were significant factors for persistent CIA. PMID- 18506621 TI - Dissonance-based Interventions for the prevention of eating disorders: using persuasion principles to promote health. AB - The limited efficacy of prior eating disorder (ED) prevention programs led to the development of dissonance-based interventions (DBIs) that utilize dissonance based persuasion principles from social psychology. Although DBIs have been used to change other attitudes and behaviors, only recently have they been applied to ED prevention. This article reviews the theoretical rationale and empirical support for this type of prevention program. Relative to assessment-only controls, DBIs have produced greater reductions in ED risk factors, ED symptoms, future risk for onset of threshold or subthreshold EDs, future risk for obesity onset, and mental health utilization, with some effects persisting through 3-year follow-up. DBIs have also produced significantly stronger effects than alternative interventions for many of these outcomes, though these effects typically fade more quickly. A meta-analysis indicated that the average effects for DBIs were significantly stronger than those for non-DBI ED prevention programs that have been evaluated. DBIs have produced effects when delivered to high-risk samples and unselected samples, as well as in efficacy and effectiveness trials conducted by six independent labs, suggesting that the effects are robust and that DBIs should be considered for the prevention of other problems, such as smoking, substance abuse, HIV, and diabetes care. PMID- 18506622 TI - Knowledge and misconceptions regarding upper respiratory infections and influenza among urban Hispanic households: need for targeted messaging. AB - BACKGROUND: To characterize knowledge and misconceptions regarding viral upper respiratory infections (URI) among urban Hispanics and identify correlates of greater knowledge. METHODS: In-home interviews conducted by trained research coordinators in 453 primarily Hispanic households (2,386 members) in northern Manhattan. RESULTS: The majority of respondents attributed URI to weather-related conditions. While most agreed that colds and flu were caused by viruses, 88.1% also agreed that they were caused by bacteria, and a small proportion reported that URI may be caused by evil eye ('mal de ojo', 7.1%) or sudden fright ('susto', 3.3%). Only 29.8% agreed that colds and flu would improve without medication; 89.9% stated that antibiotics are needed to treat viral throat infections. Most were well versed about influenza vaccination and reported that a recommendation from a healthcare provider would definitely influence them to get vaccinated. DISCUSSION: Misconceptions about the role of antibiotics in the treatment of viral infections pose considerable risk in this population. Explicit, targeted recommendations from healthcare providers regarding vaccination and judicious antibiotic use as well as media campaigns targeted to Hispanics should be effective strategies for reducing misconceptions and improving health behaviors. PMID- 18506623 TI - Looking under the Hispanic umbrella: cancer mortality among Cubans, Mexicans, Puerto Ricans and other Hispanics in Florida. AB - Cancer is the second leading cause of death among Hispanics. Most of the cancer statistics available both at the state and national levels report cancer statistics for all Hispanics as an aggregate group. The goal of this paper is to provide a population-based overview of cancer mortality among Hispanics (Cubans, Mexicans, Puerto Ricans and other Hispanics) in Florida from 1990 to 2000 and to explore the demographic diversity of this growing ethnic group. The study population consisted of Hispanics and White non-Hispanics who died from cancer. Cancer mortality rates and proportion of cancer deaths by type and age at death for the selected racial/ethnic groups were calculated. Our findings indicate that the cancer death rates of the Hispanic subgroups compared favorably with those of White non-Hispanics and that cancer rates often presented for all Hispanics mask important differences between the different ethnic subgroups that fall under the Hispanic umbrella. PMID- 18506624 TI - Influence of race, acculturation, and socioeconomic status on tendency toward overweight in Asian-American and Mexican-American early adolescent females. AB - BACKGROUND: Health disparities in chronic disease prevalence exist in the United States among racial/ethnic groups. This study explores relationships between physical, socioeconomic, and cultural characteristics of a multi-ethnic sample of early adolescent females which may assist health educators in designing programs targeting these groups. METHODS: Mexican-American and Asian-American sixth grade females (n = 144) were enrolled in Adequate Calcium Today. Physical measurements included weight, height, and BMI. Dual energy X-ray absorptiometry determined percent body fat (%BF). Socioeconomic status was determined by enrollment in free or reduced meal program (FRMP). An adapted Acculturation Rating Scale for Mexican Americans-II (ARSMA-II) measured acculturation. RESULTS: Mexican-Americans had greater height, BMI, %BF, and a greater tendency toward overweight (P < 0.01) than Asian-American. Asian-Americans were more acculturated than MA (P < 0.005), attributed to a lower ethnic orientation scale score. Within Asian-Americans, %BF was higher among FRMP participants than non-participants (P < 0.05). DISCUSSION: Income and acculturation may affect tendency toward chronic disease. PMID- 18506625 TI - Numerical experiment for ultrasonic-measurement-integrated simulation of three dimensional unsteady blood flow. AB - Integration of ultrasonic measurement and numerical simulation is a possible way to break through limitations of existing methods for obtaining complete information on hemodynamics. We herein propose Ultrasonic-Measurement-Integrated (UMI) simulation, in which feedback signals based on the optimal estimation of errors in the velocity vector determined by measured and computed Doppler velocities at feedback points are added to the governing equations. With an eye towards practical implementation of UMI simulation with real measurement data, its efficiency for three-dimensional unsteady blood flow analysis and a method for treating low time resolution of ultrasonic measurement were investigated by a numerical experiment dealing with complicated blood flow in an aneurysm. Even when simplified boundary conditions were applied, the UMI simulation reduced the errors of velocity and pressure to 31% and 53% in the feedback domain which covered the aneurysm, respectively. Local maximum wall shear stress was estimated, showing both the proper position and the value with 1% deviance. A properly designed intermittent feedback applied only at the time when measurement data were obtained had the same computational accuracy as feedback applied at every computational time step. Hence, this feedback method is a possible solution to overcome the insufficient time resolution of ultrasonic measurement. PMID- 18506626 TI - Toward a unified theory of muscle contraction. II: predictions with the mean field approximation. AB - The contractile behavior of a single half-sarcomere has been calculated from the lattice model with dimeric myosin and extensible filaments, using the model cycle with two working strokes, explicit Pi-release transitions and faster binding for the second head of the dimer. The mean-field approximation is used to generate independent state probabilities for myosin heads, assuming that the positional symmetry of actin filaments in the half-sarcomere is preserved. This model predicts absolute values of the active tension, stiffness and ATPase of fast fibers and their variation with shortening velocity, the phase-2 tension response to a length-release step and the transient tension rise during ramp stretching, in reasonable agreement with experimental data for frog muscle. It accounts for three observations beyond the reach of traditional models: (i) with elastically stiff myosin, a two-stroke model explains the rate of rapid tension recovery as a function of step size, (ii) slow Pi release from A.M.ADP.Pi after the first stroke generates the flat tension response observed after rapid recovery from a small release step, (iii) a discrete lattice model generates undamped oscillations in the isotonic length response to a force step, as observed when the sarcomeres are highly ordered. The discrete lattice also generates length dependent oscillations in the tension-length curve and the tension response to ramp shortening, which may be smoothed out if lattice symmetry is broken. PMID- 18506627 TI - Reactive oxygen species and imbalance of calcium homeostasis contributes to curcumin induced programmed cell death in Leishmania donovani. AB - Curcumin, a polyphenol compound, has been recognized as a promising anti-cancer drug. The purpose of the present study was to investigate the cytotoxicity of curcumin to Leishmania donovani, the causative agent for visceral leishmaniasis. Flow cytometric analysis revealed that curcumin induced cell cycle arrest at G2/M phase. Incubation of Leishmania promastigotes with curcumin caused exposure of phosphatidylserine to the outer leaflet of plasma membrane. This event is preceded by curcumin-induced formation of reactive oxygen species (ROS) and elevation of cytosolic calcium through the release of calcium ions from intracellular stores as well as by influx of extracellular calcium. Elevation of cytosolic calcium is responsible for depolarization of mitochondrial membrane potential (DeltaPsim), release of Cytochrome c into the cytosol and concomitant nuclear alterations that included deoxynucleotidyltransferase-mediated dUTP end labeling (TUNEL) and DNA fragmentation. Taken together, these data indicate that curcumin has promising antileishmanial activity that is mediated by programmed cell death and, accordingly, merits further investigation as a therapeutic option for the treatment of leishmaniasis. PMID- 18506628 TI - Racial differences in behavioral inpatient diagnosis: examining the mechanisms using the 2004 Florida Inpatient Discharge Data. AB - This study used the 2004 Florida Inpatient Discharge Data to examine the existence and the underlying mechanisms of the African American (AA)/white differences in behavioral inpatient diagnosis across a diverse patient population. Findings suggest that discharges of AA patients in Florida were at least twice as likely to have received a primary psychotic vs. affective or substance-related diagnosis, and this held true after between-hospital differences were controlled for. Furthermore, AA (vs. white) race was associated with a ratio of 3.3 in the population rate of hospitalization with a primary psychotic diagnosis, as compared to a ratio of 0.8 and 0.7 for affective and substance-related disorders, respectively. These findings demonstrated the wide existence of racial differences in behavioral inpatient diagnosis and the important role of clinician diagnostic behaviors as opposed to systems-level factors (such as racial differences in access to care and care-seeking behaviors) in leading to observed differences. PMID- 18506630 TI - Purification and partial characterization of lignin peroxidase from Acinetobacter calcoaceticus NCIM 2890 and its application in decolorization of textile dyes. AB - Lignin peroxidase was purified (72-fold) from Acinetobacter calcoaceticus NCIM 2890. The purified lignin peroxidase (55-65 kDa) showed dimeric nature. The maximum enzyme activity was observed at pH 1.0, between a broad temperature range of 50 and 70 degrees C, at H2O2 concentration (40 mM) and the substrate concentration (n-propanol, 100 mM). Purified lignin peroxidase was able to oxidize a variety of substrates including Mn2+, tryptophan, mimosine, L-Dopa, hydroquinone, xylidine, n-propanol, veratryl alcohol, and ten textile dyes of various groups indicating as a versatile peroxidase. Most of the dyes decolorized up to 90%. Tryptophan stabilizes the lignin peroxidase activity during decolorization of dyes. PMID- 18506629 TI - Childhood acute myeloid leukemia. AB - OPINION STATEMENT: Despite the use of intensive chemotherapy and hematopoietic stem cell transplantation, approximately one-third of children with acute myeloid leukemia (AML) still suffer relapse of their disease. It is unlikely that improvements in outcome can be achieved by further intensification of conventional chemotherapy. Instead, advances in the treatment of children with AML will require a greater understanding of the biology of the disease, with particular attention to the genetic abnormalities underlying leukemogenesis and drug resistance. Future clinical trials should include refined risk-directed therapy based on the genetics of the leukemic blasts and the patient's response to therapy. More important, we must develop alternative treatment approaches, such as agents that target specific leukemia-associated abnormalities and agents that selectively eradicate leukemic stem cells. PMID- 18506631 TI - Production of calcium-stearate by lipase using hydrogenated beef tallow. AB - Calcium-stearate has been traditionally produced by chemical methods, producing wastes and requiring high energy because of high temperature operation. To achieve enzymatic production of calcium-stearate at unfavorable conditions, i.e., pH 10 and 60 degrees C, suitable lipase was selected and reaction conditions were optimized using calcium hydroxide and hydrogenated beef tallow as substrates. Under optimum conditions, 95% of beef tallow, in 2.5 h, was converted into calcium-stearate by using commercial lipase SDL 451. Investigation of the time course reaction revealed that fatty acid was initially produced by lipase, followed by conversion into calcium-stearate. The fatty acid production rate was faster than that of the conversion into calcium-stearate at the beginning of the reaction. Alkaline pH, originating from the addition of calcium hydroxide, increased the converting reaction. This is the first report demonstrating that chemical production of calcium-stearate can be replaced by enzymatic reaction, thereby creating a cleaner process. PMID- 18506632 TI - Mediator-assisted decolorization and detoxification of textile dyes/dye mixture by Cyathus bulleri laccase. AB - Laccase from basidiomycete fungus Cyathus bulleri was evaluated for its ability to decolorize a number of reactive and acidic dyes in the presence of natural and synthetic mediators. The extent of decolorization was monitored at different mediator/dye concentrations and incubation time. Among the synthetic mediators, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) was effective at low mediator/dye ratios and resulted in 80-95% decolorization at rates that varied from 226 +/- 4 nmol min(-1) mg(-1) for Reactive Orange 1 to 1,333 +/- 15 nmol min(-1) mg(-1) for Reactive Red 198. Other synthetic mediators like 1 hydroxybenzotriazole and violuric acid showed both concentration- and time dependent increases in percent decolorization. Natural mediators like vanillin, on the other hand, were found to be less effective on all the dyes except Reactive Orange 1. Computed rates of decolorization were about twofold lower than that with ABTS. The laccase-ABTS system also led to nearly 80% decolorization for the simulated dye mixture. No clear correlation between laccase activity on the mediator and its ability to decolorize dyes was found, but pH had a significant effect: Optimum pH for decolorization coincided with the optimum pH for mediator oxidation. The treated samples were also evaluated for toxicity in model microbial systems. The laccase-mediator system appears promising for treatment of textile wastewaters. PMID- 18506633 TI - Microbial production of 1,3-propanediol by Klebsiella pneumoniae XJPD-Li under different aeration strategies. AB - The microbial production of 1,3-propanediol (1,3-PD) by Klebsiella pneumoniae XJPD-Li under different aeration strategies were investigated. In batch fermentation, the results showed that the final concentration of 1,3-PD and yield on glycerol were 13.44 g/l and 0.73 mol/mol under the anaerobic condition (N2, 0.4 vvm), 11.55 g/l and 0.62 mol/mol without aeration, and 8.73 g/l and 0.47 mol/mol under the aerobic condition (air, 0.4 vvm), respectively. Under the aerobic condition, the yield of 1,3-PD on glycerol was the lowest, while the biomass (optical density at 650 nm) was the highest among these three conditions. In the fed-batch culture, the final concentration and the yield of 1,3-PD was 60.82 g/l and 0.61 mol/mol under the anaerobic condition (N2, 0.4 vvm), 56.43 g/l and 0.53 mol/mol without aeration, and 65.26 g/l and 0.56 mol/mol under the aerobic condition. All these three conditions had good productivities of 1,3-PD, which were 3.35 g/lxh under the anaerobic condition (N2, 0.4 vvm), 3.13 g/lxh without aeration, and 3.16 g/lxh under the aerobic condition within the initial 12 h. PMID- 18506634 TI - Inhibitory effect of PACAP on caspase activity in neuronal apoptosis: a better understanding towards therapeutic applications in neurodegenerative diseases. AB - Programmed cell death, which is part of the normal development of the central nervous system, is also implicated in various neurodegenerative disorders. Cysteine-dependent aspartate-specific proteases (caspases) play a pivotal role in the cascade of events leading to apoptosis. Many factors that inhibit cell death have now been identified, but the underlying mechanisms are not fully understood. Pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to exert neurotrophic activities during development and to prevent neuronal apoptosis induced by various insults such as ischemia. Most of the neuroprotective effects of PACAP are mediated through the PAC1 receptor. This receptor activates a transduction cascade of second messengers to stimulate Bcl-2 expression, which inhibits cytochrome c release and blocks the activation of caspases. The inhibitory effect of PACAP on the apoptotic cascade suggests that selective, stable, and potent PACAP derivatives could potentially be of therapeutic value for the treatment of post-traumatic and/or chronic neurodegenerative processes. PMID- 18506636 TI - The value of computed tomography angiography in determining treatment allocation for aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Computed tomography (CT) and CT angiography (CTA) are frequently the initial imaging modalities used in the evaluation of patients with suspected aneurysmal subarachnoid hemorrhage (SAH). It remains unclear whether CTA can provide adequate information to determine best treatment modality (endovascular versus surgical) for ruptured intracranial aneurysms. METHODS: Pertinent clinical and radiological information of consecutive patients with aneurysmal SAH who underwent CTA on a 64-slice multidetector CT (MDCT) scanner were independently reviewed by five endovascular specialists. Subsequently, the interobserver reliability was calculated. RESULTS: A total of 21 consecutive patients with aneurysmal SAH detected on CTA were reviewed. Of the total of 105 reviews, in 65% a treatment allocation decision was made. Responses were, 26% either treatment; 18% endovascular only; 18% surgical only; and 3% neither treatment. In the remaining 35% it was considered that CTA images were inadequate to make a decision for treatment allocation and more information was requested. Interobserver reliability was poor between endovascular specialists (k = 0.2). The reliability was higher among endovascular/vascular neurosurgeons (k = 0.34) and physicians with >5 years of faculty experience (k = 0.55). CONCLUSION: When 64-slice MDCT angiography is used in the evaluation of aneurysmal SAH, the information obtained is adequate to determine treatment modality allocation in two-thirds of the cases. The agreement on best treatment modality varied across primary specialty, practice experience, and site of fellowship completion. PMID- 18506635 TI - Neurotrophic actions of PACAP-38 and LIF on human neuroblastoma SH-SY5Y cells. AB - The neurotrophic actions of pituitary adenylate cyclase-activating polypeptide (PACAP)-38 and leukemia inhibitory factor (LIF) were investigated in human neuroblastoma SH-SY5Y cells. Effects on differentiation were assessed through monitoring morphological changes and Western blot analysis of the expression of neuronal marker proteins. In contrast to PACAP-38, which induced a 5.5-fold increase in the number of neurite-bearing cells, LIF had no significant effect on cell morphology compared to control cells over the 4-day time course. Cells co treated with PACAP-38+LIF showed a similar increase in neurite-bearing cells compared to those treated with PACAP-38 alone. Cell morphology was similar for PACAP-38-treated and PACAP-38+LIF-co-treated cells, with the formation of bipolar neuron-like cells with long thin neurites, topped by growth cone-like structures and varicosities. SH-SY5Y cells express tyrosine hydroxylase (TH) but only low levels of the neuronal marker proteins: Bcl-2, GAP-43 and choline acetyltransferase (ChAT). Treatment of cells with PACAP-38 induced the expression of Bcl-2, GAP-43, and ChAT but did not appear to alter the expression of TH. LIF failed to induce the expression of GAP-43 and had little effect on the expression of TH, but did induce the expression of Bcl-2 and upregulated the expression of ChAT. Co-treatment with LIF had no effect on PACAP-38-induced expression of Bcl 2, GAP-43, and ChAT. Cells differentiated for 4 days with PACAP-38 or treated with LIF also displayed increased resistance to hypoxic conditions and to treatment with H2O2 and TNFalpha. The increased resistance to hypoxic conditions for PACAP-differentiated cells was blocked by the p38 MAP kinase inhibitor, SB203580, but not by the MEK1 inhibitor, PD98059. Additionally, cell proliferation assays show that LIF, but not PACAP-38, stimulates proliferation of SH-SY5Y cells, and this observed increase by LIF is not attenuated by co treatment with PACAP. Further investigation of the intracellular signaling pathways mediating the neurotrophic effects of PACAP on SH-SY5Y cells indicate that neither phospholipase C activation nor Ca2+/calmodulin-dependent kinase II (CAMKII) are involved. PMID- 18506637 TI - Recurrent takotsubo cardiomyopathy triggered by convulsive status epilepticus. AB - INTRODUCTION: Takotsubo cardiomyopathy can complicate several conditions including neurological emergencies. A few recurrent cases associated with seizures have been reported, but none of the patients had status epilepticus. The pathophysiology of takotsubo syndrome, although debated, may involve stunning of the myocardium by a catecholamine storm triggered by stress. Patients with epilepsy may be at increased risk for takotsubo syndrome, which may occur repeatedly. METHODS: We report on a postmenopausal woman with symptomatic epilepsy who experienced recurrent takotsubo cardiomyopathy triggered by convulsive status epilepticus. Brief seizures were not associated with takotsubo syndrome. The relevant literature was reviewed. RESULTS: Over a 1-year period, she experienced two episodes of convulsive status epilepticus with complete neurological recovery after treatment. Echocardiography showed latero-septo apical hypokinesia and apical ballooning. The cardiac abnormalities resolved fully and she recovered her baseline level of self-sufficiency. During the same period, she experienced several brief seizures, with no cardiac manifestations. CONCLUSION: The occurrence of takotsubo cardiomyopathy in association with convulsive status epilepticus, but not with brief seizures, supports neurogenically mediated myocardial stunning related to direct toxicity of endogenous catecholamines. Neuro-intensivists must be aware of this potentially fatal but fully reversible cardiac complication, which may be among the causes of death in patients with status epilepticus. PMID- 18506638 TI - Effect of treatment of elevated blood pressure on neurological deterioration in patients with acute intracerebral hemorrhage. AB - INTRODUCTION: Treatment of elevated blood pressure after acute intracerebral hemorrhage (ICH) is controversial. There is a risk of hematoma expansion with elevated blood pressure, and risk of ischemia with blood pressure control. This study was done to determine the effect of blood pressure control on outcome. METHODS: We retrospectively studied 122 patients with ICH. We collected 24-h blood pressure readings on all patients. The Glasgow Coma Score (GCS) at baseline and at 24 h was used to determine neurological deterioration (GCS decline > or = 2). Baseline computerized tomography (CT) scans were reviewed for hematoma volume, intraventricular hemorrhage, and location of hemorrhage. Drops in systolic blood pressure and mean arterial pressures over 24 h were divided in quartiles to determine the risk of neurological deterioration among quartiles. A logistic regression model was used to determine the association between variables of interest and neurological deterioration. RESULTS: Neurological deterioration was observed in 12 patients (10%). Baseline blood pressure and GCS were only two variables significantly different among quartiles of blood pressure drop. Multivariable adjusted analysis for these variables demonstrated significant trend toward reduced neurological deterioration with maximum blood pressure drop (systolic or mean). The risk of neurological deterioration was significantly lower in the quartile of maximum drop of systolic (odds ratio [OR] 0.02, 95% confidence interval [CI] 0.0-0.68) or mean (OR 0.03, 95% CI 0.0-0.98) blood pressure when compared to the quartile with least drop. CONCLUSION: This study supports that reduction of blood pressure in patients with acute ICH is safe and suggests that aggressive reduction might reduce the risk of neurological deterioration in first 24 h of admission. PMID- 18506639 TI - Neurogenic stunned myocardium associated with reversible posterior leukoencephalopathy syndrome. AB - INTRODUCTION: Neurogenic stunned myocardium is characterized by transient left ventricular systolic dysfunction in the absence of significant obstructive coronary artery disease. Reversible posterior leukoencephalopathy syndrome (RPLS) is characterized by transient vasogenic subcortical edema without infarction. Both syndromes are hypothesized to result from sympathetic dysregulation. We report a case of neurogenic-stunned myocardium and RPLS occurring simultaneously in a patient. METHODS: Single case report. RESULTS: We present a 55-year-old woman with a history of hypertension and chronic back pain status post spinal cord stimulator who presented with severe headache, seizure, and confusion associated with acute hypertension. Magnetic resonance imaging (MRI) of the brain revealed bilateral patchy T(2) signal hyperintensity, consistent with RPLS. Transthoracic echocardiogram (TTE) showed regional-wall motion abnormalities in the apical regions, consistent with neurogenic-stunned myocardium. The patient's TTE and MRI abnormalities resolved on follow-up studies. CONCLUSION: Neurogenic stunned myocardium and RPLS are two reversible clinical syndromes that are hypothesized to be a result of sympathetic dysregulation. Our case suggests that these two syndromes may occur together in the same patient. PMID- 18506640 TI - Diagnosis of cystic fibrosis. AB - Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that results in abnormal viscous mucoid secretions in multiple organs and whose main clinical features are pancreatic insufficiency and chronic endobronchial infection. Although it was initially defined and diagnosed based on clinical features and sweat chloride measurement, an in vivo method of assessing CFTR function, the discovery of the CFTR gene in 1989 revealed a broad spectrum of CF phenotypes associated with specific CFTR gene mutations. In this article, we will review the indications for sweat testing, alternative techniques to diagnose CF, and the approach to patients with an ambiguous or indeterminate diagnosis of CF. PMID- 18506641 TI - Workflow and methods of high-content time-lapse analysis for quantifying intracellular calcium signals. AB - Calcium ions (Ca2+) play a fundamental role in a variety of physiological functions in many cell types by acting as a secondary messenger. Variation of intracellular Ca2+ concentration ([Ca2+]i) is often observed when the cell is stimulated. However, it is a challenging task to automatically quantify intracellular [Ca2+]i in a population of cells. In this study, we present a workflow including specific algorithms for the automated intracellular calcium signal analysis using high-content, time-lapse cellular images. The experimental validations indicate the effectiveness of the proposed workflow and algorithms. We applied the workflow to analyze the intracellular calcium signals induced by different concentrations of H2O2 in the cell lines transfected by presenilin-1 (PS-1) that is known to be closely related to the familial Alzheimer's disease (FAD). The analysis results imply an important role of mutant PS-1, but not normal human PS-1 and mutant human amyloid precursor protein (APP), in enhancing intracellular calcium signaling induced by H2O2. PMID- 18506642 TI - A Java-based fMRI processing pipeline evaluation system for assessment of univariate general linear model and multivariate canonical variate analysis-based pipelines. AB - As functional magnetic resonance imaging (fMRI) becomes widely used, the demands for evaluation of fMRI processing pipelines and validation of fMRI analysis results is increasing rapidly. The current NPAIRS package, an IDL-based fMRI processing pipeline evaluation framework, lacks system interoperability and the ability to evaluate general linear model (GLM)-based pipelines using prediction metrics. Thus, it can not fully evaluate fMRI analytical software modules such as FSL.FEAT and NPAIRS.GLM. In order to overcome these limitations, a Java-based fMRI processing pipeline evaluation system was developed. It integrated YALE (a machine learning environment) into Fiswidgets (a fMRI software environment) to obtain system interoperability and applied an algorithm to measure GLM prediction accuracy. The results demonstrated that the system can evaluate fMRI processing pipelines with univariate GLM and multivariate canonical variates analysis (CVA) based models on real fMRI data based on prediction accuracy (classification accuracy) and statistical parametric image (SPI) reproducibility. In addition, a preliminary study was performed where four fMRI processing pipelines with GLM and CVA modules such as FSL.FEAT and NPAIRS.CVA were evaluated with the system. The results indicated that (1) the system can compare different fMRI processing pipelines with heterogeneous models (NPAIRS.GLM, NPAIRS.CVA and FSL.FEAT) and rank their performance by automatic performance scoring, and (2) the rank of pipeline performance is highly dependent on the preprocessing operations. These results suggest that the system will be of value for the comparison, validation, standardization and optimization of functional neuroimaging software packages and fMRI processing pipelines. PMID- 18506643 TI - Placental site trophoblastic tumor with multiple metastases and complete response to salvage BEP regimen: a case report and review of the literature. AB - Placental site trophoblastic tumor is a rare form of gestational trophoblastic disease, derived from invasive implantation site (intermediate) trophoblastic cells. It is frequently resistant to chemotherapy. Patients with metastases, however, frequently have progressive disease and die despite surgery and multiagent chemotherapy. In this case, a 24-year-old woman was referred because of intermittent vaginal bleeding episodes for 5 months following delivery. Multiple metastases in lungs, liver, kidneys, breast, pancreas, and adrenal and thyroid glands were detected. Combination therapy including surgery and multiagent chemotherapy was planned. Hysterectomy and pelvic lymph node dissection were performed. All metastatic lesions disappeared with EMA-CO treatment. However four courses of BEP regimen, salvage therapy, was performed for plateauing hCG level. Surgery and multiagent chemotherapy seem mainstay of treatment of cases having multiple metastases of PSTTs. PMID- 18506646 TI - [The German study register: always up to date on the latest developments in research]. PMID- 18506644 TI - A potent immunosuppressive retroviral peptide: cytokine patterns and signaling pathways. AB - A synthetic bioactive peptide composed of 17 amino acids (CKS-17) homologous to a highly conserved region of human and animal retroviral transmembrane envelope proteins induces not only significant immunoregulatory functions but also exhibits Th1-inhibiting properties, as described by its ability to suppress cell mediated immunity and inhibit the production of interleukin (IL) 12, IL-2, gamma interferon, and tumor necrosis factor alpha, while enhancing IL-10. An important molecular mechanism responsible for the observed cytokine profiles by CKS-17 is provided by our findings demonstrating that this small peptide activates several intracellular signaling molecules, i.e., elevates intracellular cyclic adenosine monophosphate (cAMP) levels, and induces phosphorylation of extracellular signal regulated kinase (ERK) 1 and 2, mitogen-activated protein kinase/ERK kinase (MEK), protein kinase D, Raf1, and phospholipase C gamma1 (PLCgamma1). The activation of ERK1/2 is via the PLCgamma1-protein kinase C-Raf1-MEK signaling cascade. The activation of both ERK1/2 and cAMP appears to be via a mechanism sensitive to AG879, a receptor tyrosine kinase inhibitor, but not to AG825, AG1296, or AG1478. Furthermore, phosphoinositide-3 kinase appears to mediate the CKS-17-induced activation of ERK1/2, but not of cAMP. A specific amino acid sequence as well as the dimerization of this peptide is required to confer these biological activities. The results obtained are compelling and reproducible. This highly conserved molecule may enable us to understand a basic mechanism(s) of intracellular signaling pathways, regulation of Th1/Th2 cytokines, immunosuppression, and immunologic tolerance. PMID- 18506647 TI - [Are there Good and Bad Psychotherapists?]. PMID- 18506645 TI - Autoimmune disease: is it a disorder of the microenvironment? AB - Systemic lupus erythematosus (SLE) is a common systemic autoimmune disease that involves several vital organs including the cardiovascular system, joints, and kidneys. The pathology is characterized by accumulation of autoreactive lymphocytes that attack the patients' own tissues, secretion of autoantibodies and deposition of immune complexes in vital organs. Chronic widespread inflammation is the hallmark of SLE and the target of current therapy. According to recent theories, intonating immune circuits of inflammatory cytokines and immune cells constitute highly specialized targets for SLE therapy, which nonetheless consists for the most part of anti-inflammatory medications and cytotoxic drugs. For advanced autoimmune disorders, cell therapy aiming at introducing "healthy" stem cells has been promising, keeping in mind that in its current state, stem cell therapy is reserved for the most advanced diseases refractory to traditional therapy. Ongoing studies in our laboratories examined the role of the bone marrow microenvironment, in particular, mesenchymal stem cells (MSCs) in the etiopathogenesis of SLE. Specifically, we are testing the hypothesis that, in human SLE mouse model, marrow MSCs are defective structurally and functionally. Preliminary data indicate that structural and functional defects in MSC population from an autoimmune mouse model for human SLE may contribute to this pathology and consequently present a target for cell therapy. PMID- 18506650 TI - [Therapy of snoring by electric skin stimulation]. PMID- 18506652 TI - [Malignant rhino- and otobasal mucosal melanomas--therapy and outcome]. AB - BACKGROUND: Malignant melanoma of the rhinobasal mucosa is very rare and makes up less than one percent of all malignant melanomas. Symptoms are unspecific in most cases, and patients often present with large tumours. During the past two decades, a variety of therapeutic modalities has been proposed. MATERIALS AND METHODS: In a retrospective quality assessment, we analysed the charts of fifteen consecutive patients suffering from malignant melanoma of the skull base who where treated in our department since 1993. The influence of specific surgical and adjuvant therapy on recurrence and outcome was evaluated. RESULTS: Initial symptoms were unspecific in all patients. Thus, melanoma was an accidental finding of a biopsy or sinus surgery in most patients, including all cases with amelanotic melanoma. All patients underwent surgery as the initial treatment, in 8 cases followed by adjuvant therapy. In these patients the disease specific survival was slightly better than in patients treated with surgery only. When recurrence was treated by radical mutilating surgery, this did not influence the overall prognosis. CONCLUSION: We conclude from our data and analysis of literature that the prognosis of MM has not developed favourably during the past two decades. Radiation therapy and adjuvant immuno- or chemotherapy seems to have a positive impact. Mutilating surgery is usually not indicated nor is it associated with an improvement of outcome. PMID- 18506654 TI - [Necessity of inpatient treatment in case of chronic tinnitus and mental damage due to the course of disease]. PMID- 18506655 TI - [Laryngeal intraepithelial neoplasia]. AB - Precancer (carcinoma in situ) or laryngeal intraepithelial neoplasia (LIN) is a non-invasive lesion that has genetic abnormalities, loss of cellular control functions, and some phenotypic characteristics of invasive cancer and that predicts for a substantial likelihood of developing invasive cancer. Several classifications have been proposed but none has received a total agreement. With regard to diagnosis, treatment and prognosis, these lesions differ substantially from infiltrating carcinoma. Known risk factors include cigarette smoking, viral infection with subtypes of the human papilloma virus, exposure to asbestos, and probably the gastro-oesophageal reflux disease. The diagnostic work-up usually includes indirect laryngoscopy with rigid telescopes, microlaryngoscopy and biopsies for histological evaluation. Therapeutic options include wait-and-see strategies, radiotherapy, transoral laser surgery, vocal cord stripping with cold instruments, and open partial laryngectomy. Data from the literature suggest highest local control rate with radiotherapy as initial treatment compared to other standard methods of management of dysplasia. However, transoral laser surgery can be applied repeatedly and yields excellent final results. Therefore it is now considered the treatment of choice for these lesions in the majority of patients. Local recurrences are observed more frequently than with small infiltrative carcinoma, and second primaries may arise within the upper aero digestive tract following initial treatment. Therefore systematic follow-up is recommended for these patients. PMID- 18506656 TI - [Information and consent in psychiatry and psychotherapy]. AB - OBJECTIVE: The number of medical liability court cases in Germany has more than trebled over the past 30 years. In almost every case of a therapeutic failure, the accusation is one of insufficient medical information. It was investigated which requirements information for patients has to meet in the field of psychiatry and psychotherapy. METHODS: The supreme court jurisdiction relating to patient information and consent in medicine was analysed. Legal and medical documents on the topic were also evaluated. RESULTS: Doctors' legal duty to provide information includes the areas of therapeutic information, enabling information for the patient, and information relating to costs. The burden of proof is always on the doctor, and the documentation needs to be legally safe. CONCLUSIONS: The legal basis relating to patient information and consent is consistent with the objectives of patient oriented psychiatry and psychotherapy. However, many legal requirements are exaggerated and far removed from medical practice. The specialist societies are called on to develop guidelines on how to provide information during the medical consultation. PMID- 18506657 TI - [Social representation of a psychoanalytical concept: what is the popular meaning of an "unconscious conflict"?]. AB - OBJECTIVE: A majority of the population regards unconscious conflict as a possible cause for depression or schizophrenia. We examine to what extent people associate psychoanalytical concepts with this term. METHOD: Population-based telephone survey (n = 1010), open questions about the meaning and origin of the term unconscious conflict. RESULTS: 5 % gave a definition with clearly psychoanalytical elements, another 13 % perceived an internal conflict. 24 % thought of a conflict between persons, 23 % had no answer. Regarding the origin of the term, 4 % associated Freud or psychoanalysis, 27 % psychology. For both questions, answers closer to Freudian ideas were more common in West compared to East Germany. CONCLUSION: A concretised understanding far from Freud's original conception of unconscious conflict dominates, which is even stronger in the former communist parts of Germany. Psychoanalytical terms do not necessarily carry a psychoanalytical significance with the public. PMID- 18506658 TI - [Role of the general practitioner in outpatient care for schizophrenic patients from the patients' perspective]. AB - OBJECTIVE: To explore the view of schizophrenic patients regarding the role of general practitioners (GPs) in outpatient psychiatric care. METHODS: Semi structured interviews with 20 schizophrenic patients were tape-recorded. Using the software Atlas.ti, the data were analysed according to the model of inductive category development (Mayring, 1995). RESULTS: Nearly all patients regarded the GPs as doctors "for the body" and--in contrast--the psychiatrists as doctors "for the soul". At the same time, an appointment with a GP was perceived as less embarrassing and stigmatising. Patients consulted their GP mainly because of somatic complaints or to receive follow-up prescriptions for their antipsychotic medication. They liked to get a "second opinion" regarding medical decisions by GPs. Some patients wished to have more consultation time with their GP to discuss psychological problems. CONCLUSIONS: Schizophrenic patients appreciate the distinction, and the choice, between different medical specialties. Primary care offers services with an exceptionally low threshold for schizophrenic patients because consultations with a GP are to a lesser degree perceived as embarrassing or stigmatising. PMID- 18506659 TI - Lung cancer: evolving concepts. PMID- 18506660 TI - Who is at high risk for lung cancer? Population-level and individual-level perspectives. AB - Lung cancer is the leading cause of cancer death in the world. However, there is large geographic variation internationally and within nations. Despite the fact that many causes of lung cancer have been established, cigarette smoking is the principal cause. Accounting for historical prevalence of cigarette smoking is a useful predictor of the lung cancer burden in most populations. The populations at high present risk of lung cancer can usually be predicted based on historical patterns of the prevalence of cigarette smoking, and the high-risk populations of the future can be predicted based on the current prevalence of cigarette smoking. Lung cancer rates are consistently higher among men than women and are particularly high among African American men and among those of lower socioeconomic status. At the individual level, some segments of the population (e.g., African Americans, females) have been hypothesized to have greater susceptibility to lung cancer for a given degree of cigarette smoking. Common variants in genes that encode for enzymes involved in carcinogen metabolism and detoxification and in repairing DNA damage are likely to be important determinants of interindividual susceptibility to smoking-caused lung carcinogenesis. Many lung cancer risk factors have been identified, but active cigarette smoking is the predominant cause of lung cancer and the principal marker of both high-risk populations and high-risk individuals. In the absence of cigarette smoking, lung cancer would be a rare disease. Strategies that effectively prevent youths from starting to smoke and that promote cessation among dependent smokers can transform populations from high risk to low risk. PMID- 18506661 TI - Update on screening for lung cancer. AB - Prognosis of lung cancer is markedly improved when cancers are resected in early stages (particularly in stage I). Previous investigations failed to show benefit with use of chest radiographs or sputum cytologies to screen for lung cancer among high-risk populations. More recently, computed tomography (CT) has been used as a screening technique and appears to detect lung cancer at earlier stages (e.g., stage I) compared with usual clinical practice. However, whether screening CT reduces death from lung cancer has not been clarified. This review examines the problem presented by lung cancer, the issues presented by screening, and the results of past and recent studies of lung cancer screening. PMID- 18506662 TI - Evaluation of patients with small, subcentimeter nodules. AB - The prevalence of subcentimeter pulmonary nodules is on the rise due to widespread use of multidetector row computed tomography. These lesions have created a new set of challenges for patients and their physicians. Such nodules are poorly characterized by imaging tests and are difficult to biopsy. Fortunately, the available data suggest that the vast majority of such nodules are benign in etiology. However, this increases the challenge to provide a cost effective evaluation that minimizes unnecessary radiation exposure and complications from invasive biopsies while promptly identifying malignancy when it is present. In this review, we summarize the current available literature regarding subcentimeter nodules, as well as the current recommendations regarding their investigation. PMID- 18506663 TI - Staging of non-small cell lung cancer. AB - Accurate clinical staging is critical in the evaluation of any patient with non small cell lung cancer (NSCLC). Staging for NSCLC is based on the tumor, node, metastasis (TNM) system used for most solid tumors. The clinical stage, as determined by all available clinical, radiographic, and biopsy data, provides patients and physicians with an overall assessment of disease extent and prediction of survival and enables the identification of appropriate therapies. It is thus imperative that staging be performed as accurately and comprehensively as possible while also maintaining efficiency. To do so requires an understanding of the staging system, the clinical clues that drive the staging evaluation, and a working knowledge of available noninvasive and invasive diagnostic modalities. PMID- 18506664 TI - Bronchoscopy for the diagnosis and staging of lung cancer. AB - Bronchoscopy is an invaluable tool utilized for the diagnosis, staging, and management of lung cancer. Advancements in computer technology and engineering have allowed for the emergence of newer modalities to evaluate endobronchial, parenchymal, and mediastinal pathology. Established techniques such as white light video bronchoscopy and its ancillary procedures (forceps biopsy, brush biopsy, bronchoalveolar lavage, bronchial washings, and transbronchial needle aspiration) are discussed here, with their accuracy described in relation to tumor location, size, and type. Newer technologies such as autofluorescence bronchoscopy, narrow band imaging, endoscopic ultrasound, endobronchial ultrasound, electromagnetic navigation, optical coherence tomography, and confocal fluorescent laser microscopy are introduced and put into perspective. Special emphasis has been placed on their role in the early detection and staging of lung cancer. Some technology requires further study to delineate its role in the disease, whereas other modalities are emerging as the new gold standard in evaluation of lung cancer. The future holds great promise with further miniaturization of equipment and improvements in computer processing power that may allow for in vivo pathological evaluation of abnormal tissue. PMID- 18506665 TI - Lung cancer: preoperative pulmonary evaluation of the lung resection candidate. AB - Lung resection provides the best chance of cure for individuals with early-stage non-small-cell lung cancer. Lung resection will lead to a decrease in lung function. The population that develops lung cancer often has concomitant lung disease and a reduced ability to tolerate further losses in lung function. The goal of the preoperative pulmonary assessment of individuals with resectable lung cancer is to identify those individuals whose short- and long-term morbidity and mortality would be unacceptably high if surgical resection were to occur. Pulmonary function measures such as the forced expiratory volume in 1 second (FEV1) and the diffusing capacity for carbon monoxide (DLCO) are useful predictors of postoperative outcome. When lung function in not normal, the prediction of postoperative lung function from preoperative results, and the assessment of exercise capacity, can help to clarify the risks. Testing algorithms and published guidelines help to direct the order of testing. Interventions that allow us to consider resection outside of the guidelines, or to maximize the potential to tolerate lung resection, help us to offer potentially curative treatment to as many as possible. Knowledge of alternative treatment options will help to guide those deemed unfit for resection. PMID- 18506666 TI - Advances in radiotherapy for lung cancer. AB - The past decade has seen steady advances in the field of radiation therapy, impacting definitive treatment options for early and advanced tumors, as well as identifying further roles for adjuvant and palliative therapy. The widespread availability of (18)F-fluoro-2-deoxy-D-glucose positron emission tomographic (FDG PET) imaging has improved staging, and better defines a population that will benefit from aggressive local therapy. Technological advances in radiation treatment planning allow for less normal lung irradiation, allowing increasing dose and decreasing normal tissue toxicity. Ongoing clinical investigations in the role of radiosurgery for early lung tumors, postoperative radiation of locally advanced non-small-cell lung cancer (NSCLC), and prophylactic cranial irradiation in small-cell lung cancer (SCLC) help to identify patients who may benefit from treatment. PMID- 18506667 TI - Targeted therapy in advanced non-small-cell lung cancer. AB - Molecularly targeted therapies have recently expanded the options available for patients with advanced non-small-cell lung cancer (NSCLC). Two cancer cell pathways in particular have been exploited, the epidermal growth factor receptor (EGFR) and the vascular endothelial growth factor (VEGF) pathway. The former has emerged as a key regulator of cancer cell proliferation and invasion, and several EGFR inhibitors have been developed. Erlotinib, a small-molecule inhibitor of the EGFR intracellular tyrosine kinase, has been found to improve survival compared with placebo in previously treated patients with advanced NSCLC and is Food and Drug Administration (FDA)-approved in this setting. Clinical and molecular predictors of response to erlotinib, such as a history of never smoking and EGFR gene mutation or amplification, are presently being evaluated to select patients for earlier therapy with erlotinib. Additional EGFR inhibitors are also being examined in randomized trials. The VEGF pathway, a key mediator of angiogenesis, has become an attractive target in multiple malignancies, including lung cancer. Bevacizumab, a monoclonal antibody to VEGF, received FDA approval for use in advanced non-squamous-cell NSCLC in 2006 after a phase III trial reported a significant survival advantage when bevacizumab was added to standard first-line chemotherapy. Small-molecule inhibitors of the VEGF receptor tyrosine kinase, such as sunitinib and sorafenib, have also shown promise in phase II trials and are being further investigated in phase III studies. Because preclinical data suggest a synergistic effect when VEGF and EGFR inhibitors are combined, the concurrent use of erlotinib and bevacizumab has additionally been evaluated in a phase II trial, with encouraging early results suggesting at least equivalent activity to standard salvage chemotherapy, with less toxicity. Several other novel agents are being examined, including inhibitors of histone deacteylases and the 26S proteosome. Research efforts are currently focusing on tailoring such therapies according to predictive clinical and molecular markers. PMID- 18506668 TI - Complications of lung cancer treatment. AB - In most cases lung cancer is incurable, but treatment is prolonging life for many and sustaining quality of life. Inevitably, disease-related symptoms develop with disease progression, and it can be difficult to differentiate these from treatment-induced complications. This is particularly true for pulmonary complications because tumor progression occurs most frequently in the lungs, and separating the effects of the disease from those induced by treatment (chemotherapy, radiotherapy) is often very difficult. This chapter identifies the main complications around surgical resection of lung cancer, highlighting the importance of expert postoperative care. For palliative treatments such as radiotherapy and chemotherapy, complications are related to the planned intensity of proposed therapy, the performance status of the patients, and patient age. Treatment of the elderly sufferer, now representing almost 50% of new cases, is poorly researched, but therapy-related complications are commoner in those aged over 70 years by ~20% when compared with their younger counterparts. Even during palliative care great attention has to be taken to minimize side effects of commonly used medications. PMID- 18506670 TI - Predictors of breast cancer-related distress following mammography screening in younger women on a family history breast screening programme. AB - OBJECTIVE: This longitudinal study investigated pre-screening factors that predicted breast cancer-specific distress among 1286 women who were undergoing annual mammography screening as part of a UK programme for younger women (i.e., under 50) with a family history of breast cancer. METHODS: Women completed questionnaires one month prior to screening, and one and six months after receiving screening results. Factors measured were breast cancer worry, perceived risk, cognitive appraisals, coping, dispositional optimism, and background variables relating to screening history and family history. RESULTS: Pre screening cancer worry was the most important predictor of subsequent worry, explaining 56/61% and 54/57% of the variance at one and six months follow-up, respectively. Other salient pre-screening predictors included high perceived risk of breast cancer, appraisals of high relevance and threat associated with the family history, and low perceived ability to cope emotionally. Women who had previously been part of the screening programme and those with a relative who had recently died from breast cancer were also vulnerable to longer-term distress. A false positive screening result, pessimistic personality, and coping efforts relating to religion and substance use predicted outcomes of screening at one month follow-up, but were not predictive in the longer-term. CONCLUSION: Early intervention to ameliorate high levels of cancer-related distress and negative appraisals would benefit some women as they progress through the familial breast screening programme. PMID- 18506671 TI - Preliminary results of a longitudinal study of changes in cognitive function in breast cancer patients undergoing chemotherapy with doxorubicin and cyclophosphamide. AB - OBJECTIVES: Recent studies suggest that standard dose chemotherapy for breast cancer may cross the blood-brain barrier. However, the evidence for chemotherapy induced cognitive impairments in breast cancer patients is inconsistent. The purposes of this study in a sample of newly diagnosed patients with breast cancer were to (1) evaluate cognitive function prior to the administration of chemotherapy; (2) assess changes in cognitive function over time; and (3) evaluate potential relationships between cognitive function and anxiety, depression, fatigue, hemoglobin level, menopausal status, and perception of cognitive function. METHODS: Thirty women with breast cancer completed neuropsychological testing before the initiation of chemotherapy and after four cycles of doxorubicin and cyclophosphamide. Descriptive statistics were used to summarize sample characteristics, and paired t-tests were carried out to evaluate for changes in neuropsychological test scores prior to and following completion of chemotherapy. Linear mixed model analyses were used to determine whether significant changes in neuropsychological test scores remained after controlling for anxiety, depression, fatigue, hemoglobin level, menopausal status, and perceived cognitive function. RESULTS: Significant decreases in visuospatial skill (p<0.001) and total cognitive scores (p=0.001) were found following chemotherapy. In addition, a significant improvement was found in executive function (p=0.014). Of note, these changes remained significant even after controlling for anxiety, depression, fatigue, hemoglobin level, menopausal status, and perceived cognitive function. CONCLUSIONS: Data from this study supported the hypothesis that chemotherapy may have a negative impact on select domains of cognitive function. PMID- 18506672 TI - Residues of 14C-metsulfuron-methyl in Chinese paddy soils. AB - BACKGROUND: Metsulfuron-methyl is widely used for controlling many annual grasses and broadleaf weeds in cereal crops. Nonetheless, increasing evidence has demonstrated that even extremely low levels of metsulfuron-methyl residues in soil can be toxic to subsequent crops or non-target organisms. The behavior of herbicides in soils is mostly related to their residual forms. The intent of the present study was to investigate the dynamics of extractable residues (ERs) and non-extractable residues (NERs) of (14)C-metsulfuron-methyl in twelve Chinese paddy soils and their relationships to soil properties. RESULTS: ERs decreased gradually after application, whereas NERs increased rapidly during the initial 28 days, and gradually decreased thereafter. ERs and NERs were respectively 10.1 67.9% and 5.6-28.7% of applied radioactivity in soils at 224 days after application. ERs correlated positively with soil pH and silt fractions, and negatively with microbial biomass carbon (MBC) and clay fractions, but the opposite was observed for NERs. CONCLUSION: Both ERs and NERs may be present in the soil at the time of planting following rice crops, and the risk of phytotoxic effects needs to be considered. Soil pH, MBC and clay/silt fractions were the main factors in affecting the amounts of both ERs and NERs of metsulfuron-methyl in the tested soils. PMID- 18506673 TI - Buprofezin susceptibility survey, resistance selection and preliminary determination of the resistance mechanism in Nilaparvata lugens (Homoptera: Delphacidae). AB - BACKGROUND: Buprofezin has been used for many years to control Nilaparvata lugens (Stal). Assessment of susceptibility change in the insect is essential for maintaining control efficiency and resistance management. RESULTS: Eleven-year surveys showed that most field populations were susceptible before 2004. However, substantially higher levels of resistance (up to 28-fold) were found in most of the rice fields in China after 2004. A field population was collected and periodically selected for buprofezin resistance in the laboratory. After 65 generations (56 were selected), the colony successfully obtained 3599-fold resistance to buprofezin. Synergism tests showed that O,O-diethyl-O-phenyl phosphorothioate (SV1), piperonyl butoxide (PBO) and diethyl maleate (DEM) increased buprofezin toxicity in the resistant strain by only 1.5-1.6 fold, suggesting that esterases, P450-monooxygenases and glutathione S-transferases had no substantial effect on buprofezin resistance development. CONCLUSION: The results from this study indicate that N. lugens has the potential to develop high resistance to buprofezin. A resistance management program with rotation of buprofezin and other pesticides may efficiently delay or slow down resistance development in the insect. Further investigation is also necessary to understand the resistance mechanisms in N. lugens. PMID- 18506674 TI - Response of citrus peelminer Marmara gulosa Guillen and Davis (Lepidoptera: Gracillariidae) stages to various insecticides. AB - BACKGROUND: Egg and larval stages of citrus peelminer Marmara gulosa Guillen and Davis infesting zucchini squash Cucurbita pepo L. var 'Revenue' were exposed in the laboratory to a wide range of insecticide classes used in California citrus (organophosphate, carbamate, pyrethroid, neonicotinoid, insect growth regulator, fermentation products and sulfur) to determine peelminer response with and without a non-ionic adjuvant or oil. RESULTS: All of the insecticides tested except sulfur reduced egg hatch and mine initiation. When the larval stage was treated, only buprofezin failed to reduce larval and pupal development. Cyfluthrin and spinosad, with the addition of adjuvant or oil, and abamectin combined with oil allowed no survivors at 7 days after treatment (DAT). The slower-acting insect growth regulators pyriproxyfen and diflubenzuron applied in combination with adjuvant or oil allowed no survival of peelminer 21 DAT. A field trial showed 62.6-68.5% reduction in mined citrus fruit after two applications of diflubenzuron and no significant improvement of control with the addition of cyfluthrin. CONCLUSION: Bioassays indicate that M. gulosa is relatively susceptible to a wide range of insecticide classes in the laboratory. Lack of efficacy experienced in field trials would then be due to characteristics of the fruit or canopy that limit coverage, rather than to the effectiveness of the insecticides. PMID- 18506675 TI - Differential correlates of reactive and proactive aggression in Asian adolescents: relations to narcissism, anxiety, schizotypal traits, and peer relations. AB - This study investigated relationship between reactive and proactive aggression, and narcissism, anxiety, schizotypal traits, and interpersonal relations in a sample of 698 Asian adolescents from Grades 7 to 9. Proactive aggression was found to be significantly associated with narcissism, whereas reactive aggression was significantly associated with anxiety, schizotypal traits, and poor interpersonal relations. Study findings provide support from a cross-cultural standpoint for differential correlates of reactive and proactive aggression and represent an initial attempt to illustrate the generalizability of existing findings on the distinction between the two subtypes in an Asian context. Implications for theory building of the reactive-proactive aggression typology are discussed. PMID- 18506676 TI - Contingent responses of mothers and peers to indirect and direct aggression in preschool and school-aged children. AB - The primary goal of the study was to determine whether mother and peer's responses to direct and indirect aggression would contribute to children's use of direct and indirect aggression. Using adaptations of the Direct and Indirect Aggression Scale, a multi-informant strategy, and a sample of disadvantaged families, data were collected from 296 mothers of children ages 4-11, 237 children ages 6-11, and 151 teachers of those children. Mothers and peers were reported to react more harshly in response to direct aggression compared with indirect aggression, and higher rates of direct aggression were associated with reduced popularity. These findings were seen as being consistent with the hypothesis that different forms of aggression result in differential responding by mothers and peers, as well as the notion that direct aggression is a higher cost option than indirect aggression. Results also replicated previous findings that boys tend to use physical aggression more than girls, but girls use indirect aggression more than boys. Finally, low IQ was correlated with higher direct aggression in girls but had no relation with aggression in boys. PMID- 18506677 TI - Are the effects of Unreal violent video games pronounced when playing with a virtual reality system? AB - This study was conducted to analyze the short-term effects of violent electronic games, played with or without a virtual reality (VR) device, on the instigation of aggressive behavior. Physiological arousal (heart rate (HR)), priming of aggressive thoughts, and state hostility were also measured to test their possible mediation on the relationship between playing the violent game (VG) and aggression. The participants--148 undergraduate students--were randomly assigned to four treatment conditions: two groups played a violent computer game (Unreal Tournament), and the other two a non-violent game (Motocross Madness), half with a VR device and the remaining participants on the computer screen. In order to assess the game effects the following instruments were used: a BIOPAC System MP100 to measure HR, an Emotional Stroop task to analyze the priming of aggressive and fear thoughts, a self-report State Hostility Scale to measure hostility, and a competitive reaction-time task to assess aggressive behavior. The main results indicated that the violent computer game had effects on state hostility and aggression. Although no significant mediation effect could be detected, regression analyses showed an indirect effect of state hostility between playing a VG and aggression. PMID- 18506678 TI - Evaluation of headspace-gas chromatography/mass spectrometry for the analysis of benzene in vitamin C drinks; pitfalls of headspace in benzene detection. AB - Recently, there have been reports regarding the presence of benzene in vitamin C drinks. This is caused by sodium benzoate and ascorbic acid (vitamin C), which can react together to induce benzene formation. While the headspace gas chromatography method is well known for the detection of benzene, there could be pitfalls in the process of benzene extraction. This study was performed to check if benzene could be generated under high-temperature incubation conditions. As a result, the amount of benzene detected by headspace-gas chromatography/mass spectrometry (HSGC/MS) was affected by temperature changes. As the temperature of the sample vial was increased, newly generated benzene from the headspace also increased, causing false-positive determination of benzene. Although 80 degrees C is generally accepted for the temperature of headspace sample vials, lower temperatures, such as 40 degrees C, minimize the false-positive identification of benzene. Considering that this minimization allows benzene to be quantified at around 5 ppb, this lower temperature should definitely be considered since benzene, which is formed in sodium benzoate, can appear in vitamin C drinks under certain circumstances. The proposed analysis method of benzene in vitamin C drinks by HSGC/MS at 40 degrees C is an accurate and universal method for the monitoring of benzene without false-positive identification. PMID- 18506679 TI - The role of alternative specimens in toxicological analysis. AB - The use of alternative specimens in the field of toxicology was first described in 1979, when hair analysis was used to document chronic drug exposure. Since then, the use of these 'alternative' samples has gained tremendous importance in forensic toxicology, as well as in clinic toxicology, doping control and workplace drug testing. It is not surprising, therefore, that a large number of papers dealing with the determination of several classes of drugs in saliva, sweat, meconium and hair have been published ever since, owing to the fact that chromatographic equipment is becoming more and more sensitive, mass spectrometry (and tandem mass spectrometry) being the most widely used analytical tool, combined with gas or liquid chromatography. 'Alternative' specimens present a number of advantages over the 'traditional' samples normally used in toxicology (e.g. blood, urine and tissues), namely the fact that their collection is not invasive, their adulteration is difficult, and they may allow increased windows of detection for certain drugs. The main disadvantage of this kind of samples is that drugs are present in very low concentrations, and therefore high-sensitivity techniques are required to accomplish the analysis. This paper reviews a series of publications on the use of alternative specimens, with special focus on the main analytical and chromatographic problems that these samples present, as well on their advantages and disadvantages over traditional samples in documenting drug exposure. PMID- 18506680 TI - Optimization, validation and application of a method for the determination of trichloroethylene in rat plasma by headspace solid-phase microextraction gas chromatography mass spectrometry. AB - Trichloroethylene (TCE) is a small halogenated compound that has been used extensively as a metal degreaser and a solvent for the past 100 years. As a result of its widespread use, TCE can be found in the groundwater of about one third of the hazardous waste sites on the United States Environmental Protection Agency's National Priorities List. Human exposure to TCE in the environmental media is of concern because TCE has been found to be carcinogenic in laboratory animals. This paper describes the development and validation of a HS-SPME-GC/MS method for determination of TCE in rat plasma. The effects of different parameters such as sample volume, extraction and desorption conditions, fiber positions and salt addition were investigated and optimized. The method is rapid, simple, sensitive and requires a very small sample volume. The lower limit of quantitation was 0.25 ng/mL and correlation coefficient (r(2)) values for the linear range of 0.25-100 ng/mL were 0.996 or greater. The precision and accuracy for intra-day and inter-day were better than 8.0%. This validated method was successfully applied to study the toxicokinetic behavior of TCE following low levels of oral administration. PMID- 18506682 TI - Liquid chromatography tandem mass spectrometry method for the quantification of clonidine with LLOQ of 10 pg/mL in human plasma. AB - A sensitive high-performance liquid chromatography-positive ion electrospray tandem mass spectrometry method was developed and validated for the quantification of clonidine in human plasma. Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse-phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M + H](+) ions, m/z 230 to 44 for clonidine and m/z 254 to 44 for the internal standard. The assay exhibited a linear dynamic range of 10-2000 pg/mL for clonidine in human plasma. The lower limit of quantification was 10 pg/mL with a relative standard deviation of less than 6.8%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 2.5 min for each sample made it possible to analyze more than 250 human plasma samples per day. The validated method was successfully used to analyze human plasma samples for application in pharmacokinetic studies. PMID- 18506683 TI - Liquid chromatographic methods for biotransformation studies of ochratoxin A. AB - Liquid chromatographic methods were used for the detection of ochratoxin A (OTA) and its metabolites ochratoxin alpha (OTalpha), 10-hydroxy OTA (10-OHOTA), 4R hydroxy OTA (4R-OHOTA) and the ethyl ester of OTA (OTC) in in vitro samples, obtained with Caco-2 cell culture experiments and in in vivo urine samples from sheep. A high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) and a liquid chromatography-tandem mass spectrometry (LC MS/MS) method were developed and validated for the detection of OTA and its metabolites OTalpha, 10-OHOTA, 4R-OHOTA and OTC, which was used as internal standard. The LOD/LOQ values for OTalpha, 4R-OHOTA and OTA were 0.63/2.11, 0.99/3.31 and 0.84/2.81 microg/L, respectively, for the HPLC-FLD method and 0.98/3.28, 1.11/3.72 and 0.88/2.96 microg/L, respectively for the LC-MS/MS method. Within-day and between-day precision were both <12% for the HPLC-FLD method, and <10% for the LC-MS/MS method. The recovery of OTA and its metabolites ranged between 71 and 111% for the HPLC-FLD method and between 79 and 110% for the LC-MS/MS method. In the first experiment only OTA was added to the Caco-2 cells while in the second experiment 3-methylcholanthrene (3MC) was also present in the cell culture systems. Besides OTA, which was recovered in all the samples, an unknown compound was also observed in the second experiment. When 3MC was added, the results showed that the OTA concentration in the basolateral samples was decreased by 50%. The methods were also implemented for the analysis of urine samples of sheep, fed increasing amounts of OTA. With the HPLC-FLD method it could be concluded that the concentration of OTA and OTalpha increased according to ingested amounts of OTA, with OTalpha being the most abundant compound. The results obtained with the LC-MS/MS method confirmed these results. PMID- 18506684 TI - Relational aggression and victimization in gay male relationships: the role of internalized homophobia. AB - This article presents two studies that are the first to examine relational aggression and relational victimization in gay male peer relationships. A qualitative pilot study provides a strong rationale for a subsequent empirical investigation of 100 young adult, self-identified gay males. Results of both studies demonstrate that relational aggression and relational victimization are common experiences in gay male relationships. They also reveal forms of relational aggression and victimization that appear to be unique to gay males (e.g., outing). Results of the empirical study found significant relations between engaging in relational aggression against gay males and experiencing relational victimization and between experiencing relational victimization and internalized homophobia. However, there was no significant correlation between internalized homophobia and engaging in relational aggression. A multiple regression analysis found that experiencing relational victimization was correlated more strongly with the combination of engaging in relational aggression and internalized homophobia together than with relational aggression alone. Results are discussed within the framework of Allport's "traits due to victimization" theory and Meyer's theory of "minority stress." Implications for the prevention of relational aggression/victimization in gay male relationships are offered. PMID- 18506685 TI - Lysophosphatidic acid induces ovarian cancer cell dispersal by activating Fyn kinase associated with p120-catenin. AB - Lysophosphatidic acid (LPA), known as the "ovarian cancer activating factor," is a natural phospholipid involved in important biological functions, such as cell proliferation, wound healing and neurite retraction. LPA causes colony dispersal in various carcinoma cell lines by inducing morphological changes, including membrane ruffling, lamellipodia formation, cell-cell dissociation and single cell migration. However, its effects on cell-cell dissociation and cell-cell adhesion of ovarian cancer cells have not been studied. In our study, we showed that LPA induced sequential events of intercellular junction dispersal and "half-junction" formation in ovarian cancer SKOV3 cells and that Src-family kinases were involved in both processes, since the effects were abolished by the selective tyrosine kinase inhibitor PP2. LPA induced rapid and transient activation of Src family kinases, which were recruited to cell-cell junctions by increasing the association with the adherens junction protein p120-catenin. We identified the Src family kinase, Fyn, as the key component associated with p120-catenin after LPA stimulation in SKOV3 cells. Our study provides evidence that LPA induces junction dispersal in ovarian cancer SKOV3 cells by activating the Src family kinase Fyn and increasing its association with p120-catenin at the cell-cell junction. PMID- 18506686 TI - To err is human; to follow the SOP divine. PMID- 18506687 TI - Blood transfusion, anesthesia, surgery and risk of non-Hodgkin lymphoma in a population-based case-control study. AB - The incidence of NHL has increased dramatically since at least the 1950s, and during this timeframe there has been a major increase in the use of blood transfusions, invasive surgical procedures and anesthesia, all of which can impact immune function. We evaluated these factors with NHL risk in a population based study of 759 cases and 589 frequency-matched controls. Risk factor data were collected during in-person interviews. Unconditional logistic regression was used to estimate ORs and 95% CIs, adjusted for the matching factors. History of transfusion was associated with a 26% higher risk of NHL (95% CI 0.91-1.73), and the elevated risk was specific to transfusions first given 5-29 years before the reference date (OR = 1.69; 95% CI 1.08-2.62) and transfusions given for a medical condition (OR = 2.09; 95% CI 1.03-4.26). The total number of surgeries and dental procedures (OR = 1.53 for 26+ surgeries compared to 0-6; 95% CI 1.02-2.29) and to a lesser extent the total number of exposures to general or local/regional anesthesia (OR = 1.35 for 24+ times compared to 0-6; 95% CI 0.91-2.02) were positively associated with risk of NHL. Inclusion of transfusion and surgery or transfusion and anesthesia in the same model did not attenuate these associations. All results were broadly consistent for both DLBCL and follicular subtypes. Blood transfusions were associated with NHL risk, but appear to be a marker for underlying medical conditions. Multiple surgical procedures and/or repeated administration of anesthesia have not been previously reported to be associated with risk of NHL and these exposures warrant further evaluation. PMID- 18506688 TI - Breast cancer expression of CD163, a macrophage scavenger receptor, is related to early distant recurrence and reduced patient survival. AB - Cells of the monocyte/macrophage lineage are important for tumour cell migration, invasion and metastasis. Fusion between macrophages and cancer cells in animal models in vitro and in vivo causes hybrids with increased metastatic potential. Primary breast cancer cells were characterized for macrophage antigens to test if phenotypic resemblance to macrophages is related to early distant recurrence. Immunostaining for CD163, MAC387 and CD68 was performed in a breast cancer tissue micro array from 127 patients consequently followed up for a median of 13 years. Tumour-associated macrophages expressed all 3 antigens. The breast cancers expressed CD163 to 48%, MAC387 to 14% while CD68 was not expressed. TGF-beta staining intensity was positively related to both CD163 and MAC387 expression. Expression of CD163 in the cancer cells was compared to their DNA ploidy, Nottingham Histological Grade, TNM-stage, node state, presence of estrogen receptors and occurrence of distant metastases and survival. Cancers of a more advanced histological grade expressed CD163 to a higher extent. Cells expressing MAC387 were more common in cancers with a high proportion of CD163 positive cells. Multivariate analysis showed that expression of the macrophage antigen CD163 in breast cancer cells has a prognostic impact on the occurrence of distant metastases and reduced patient survival time. PMID- 18506689 TI - KIT overexpression induces proliferation in astrocytes in an imatinib-responsive manner and associates with proliferation index in gliomas. AB - Activating gene mutations, gene amplifications and overexpressed proteins may be useful as targets for novel therapies. Alterations at chromosome locus 4q12 are associated with gliomas and the region harbors the receptor tyrosine kinase gene KIT, which is frequently amplified in gliomas, and also overexpressed in a subset of gliomas. KIT and its ligand stem cell factor are widely expressed in embryonic and adult mouse brain, and they play a role in many signal transduction pathways involved in cellular proliferation, differentiation and cancer cell metastasis. However, the function of KIT in gliomagenesis or disease progression remains unresolved as well as its role in neural and brain tumor development. In this study, we utilized lentivirus-mediated gene transfer to deliver the KIT gene into mouse astrocytes. The growth properties of KIT overexpressing cells were analyzed using several in vitro functional assays. The effect of receptor tyrosine kinase inhibitor imatinib on astrocyte growth was also investigated. Our results indicate that overexpression of KIT in mouse astrocytes promotes cell proliferation, and the increased proliferation is partly inhibited by imatinib treatment. Furthermore, KIT overexpression induces phenotypic changes in the cells suggesting that KIT may play a role in astrocyte growth regulation. PMID- 18506690 TI - Integrated gene copy number and expression microarray analysis of gastric cancer highlights potential target genes. AB - We performed an integrated array comparative genomic hybridization (aCGH) and expression microarray analysis of 8 normal gastric tissues and 38 primary tumors, including 25 intestinal and 13 diffuse gastric adenocarcinomas to identify genes whose expression is deregulated in association with copy number alteration. Our aim was also to identify molecular genetic alterations that are specific to particular clinicopathological characteristics of gastric cancer. Distinct molecular genetic profiles were identified for intestinal and diffuse gastric cancers and for tumors obtained from 2 different locations of the stomach. Interestingly, the ERBB2 amplification and gains at 20q13.12-q13.33 almost exclusively discriminated intestinal cancers from the diffuse type. In addition, the 17q12-q25 gain was characteristic to cancers located in corpus and the 20q13.12-q13.13 gain was more common in the antrum. Statistical analysis was performed using integrated copy number and expression data to identify genes showing differential expression associated with a copy number alteration. Genes with the highest statistical significance included ERBB2, MUC1, GRB7, PPP1R1B and PPARBP with concomitant changes in copy number and expression. Immunohistochemical analysis of ERBB2 and MUC1 on a tissue microarray containing 78 independent gastric tissues showed statistically significant differences (p < 0.05 and <0.001) in immunopositivity in the intestinal (31 and 70%) and diffuse subtypes (14 and 41%), respectively. In conclusion, our results demonstrate that intestinal and diffuse type gastric cancers as well as cancers located in different sites of the stomach have distinct molecular profiles which may have clinical value. PMID- 18506691 TI - High preoperative plasma TIMP-1 is prognostic for early relapse in primary breast carcinoma. AB - TIMP-1 is a natural inhibitor of extracellular matrix degrading enzymes called matrix metalloproteinases. In addition to its capacity to inhibit matrix degradation, TIMP-1 has been shown to promote cell growth and inhibit apoptosis. The expression of TIMP-1 in tumor tissue, as well as in circulating blood, has therefore been shown to associate with worsened survival in several malignancies. In our study, a prospective series of 213 patients with primary breast carcinoma was assessed. Circulating pre- and postoperative TIMP-1 levels were assayed using enzyme-linked immunosorbent assay analysis. It was shown that high preoperative plasma TIMP-1 was a powerful predictor of systemic early relapse in breast carcinoma, with HR 8.1 (95% CI 1.8-37.6) (p = 0.007) as a log-transformed continuous variable in Cox regression univariate analysis. It was shown to be independent of, and superior to, nodal status as a prognostic variable in multivariate analysis, and not associated with any known prognostic clinicopathological parameters. Kaplan-Meier analysis showed that the patients belonging to the highest quartile of circulating TIMP-1 levels had a worsened recurrence-free survival of 79% compared to 94% RFS among patients in the lower quartiles (p = 0.016). The postoperative levels of circulating plasma TIMP-1 were not found to be prognostic for relapse. In conclusion, preoperative plasma TIMP-1 was found to be a powerful prognostic factor for early systemic relapse in primary breast carcinoma. PMID- 18506692 TI - Risk of different histological types of postmenopausal breast cancer by type and regimen of menopausal hormone therapy. AB - In a large population-based case-control study in Germany, including 3,464 breast cancer cases aged 50-74 at diagnosis and 6,657 population based and frequency matched controls, we investigated the effects of menopausal hormone therapy (HT) by type, regimen, timing and progestagenic constituent on postmenopausal breast cancer risk overall and according to histological type. Data were collected by face-to-face interviews. Logistic and polytomous logistic regression analysis were used to estimate odds ratios (OR) and 95%-confidence intervals (95% CI). Risk of invasive breast cancer was significantly elevated in current users (OR, 1.73, 95% CI, 1.55-1.94) and heterogeneous by histological type (p < 0.01), being more than 2-fold higher for lobular and tubular than for ductal cancer. Risks for current users varied significantly by type and regimen of HT, with ORs per year of use of 1.05 (95% CI, 1.04-1.06) for continuous combined estrogen-progestagen, 1.03 (95% CI, 1.02-1.04) for cyclical EP and 1.01 (95% CI, 1.00-1.03) for estrogen-only therapy. No statistically significant increase in risk was observed after 5 years of cessation of HT use for any histological type. Analyses of progestagenic content by regimen revealed a significantly higher risk for continuously administered norethisterone- or levonorgestrel-derived progestagens than for continuously administered progesterone-derived progestagens (OR, 2.27, 95% CI, 1.98-2.62 vs. 1.47, 95% CI, 1.12-1.93, respectively, p = 0.003), which may be explained by dose rather than type of progestagen. These data suggest that the risks associated with menopausal HT differ by type and regimen of HT and histological type of breast cancer and may vary by progestagenic component, depending on the effective dose. PMID- 18506693 TI - Prediction of the immunogenic potential of frameshift-mutated antigens in microsatellite instable cancer. AB - Microsatellite instable (MSI) cancers express frameshift-mutated antigens, the C terminal polypeptides of which are foreign to the immune system. Consequently, these antigens constitute a unique pool of tumor-specific antigens that can be exploited for patient diagnosis and selective, immune-mediated targeting of cancers. However, other than their sequence, very little is known about the characteristics of the majority of these proteins. We therefore developed a methodology for predicting their immunogenic behavior that is based on a gene expression system, in which each of the proteins was fused to a short C-terminal polypeptide comprising two epitopes that can be readily detected by T-cells and antibodies, respectively. In this manner, accumulation of the antigens and processing of peptides derived thereof into MHC can be monitored systematically. The antigens, which accumulate in the cells in which they are synthesized, are of primary interest for cancer immunotherapy, because peptide epitopes derived thereof can be presented by dendritic cells in addition to the tumor cells themselves. As a result, these antigens constitute the best targets for a coordinated immune response by both CD8+ and CD4+ T-cells, which increases the likelihood that tumor-induced immunity would be detectable against these antigens in cancer patients, as well as the potential value of these antigens as components of anticancer vaccines. Our data indicate that, of 15 frameshift mutated antigens examined in our study, 4 (TGFbetaR2-1, MARCKS-1, MARCKS-2 and CDX2-2) are of primary interest, and 4 additional antigens (TAF1B-1, PCNXL2-2, TCF7L2-2 and Baxalpha+1) are of moderate interest for further tumor immunological research. PMID- 18506694 TI - Mountain gorilla research: the risk of disease transmission relative to the benefit from the perspective of ecosystem health. PMID- 18506695 TI - Kick: constraining a stochastic search procedure with molecular fragments. AB - An extension of the Kick program developed by Bera et al. (J Phys Chem A 2006, 110, 4287) is described in which chemically sensible molecular fragments are used in an automated stochastic search algorithm. This results in a vastly reduced region of the potential energy surface which can be explored very quickly. We present use of this modified algorithm to the search for low-lying isomers, and we present candidates for the global energy minimum, for a range of chemical systems. We highlight the usefulness of this procedure for exploring reactions of molecules with transition metal clusters and to the microsolvation of a small dipeptide. PMID- 18506696 TI - Determination of sodium cromoglycate in human plasma by liquid chromatography with tandem mass. AB - A sensitive and selective liquid chromatography-tandem mass spectrometric (LC MS/MS) method was developed and validated for the determination of sodium cromoglycate (SCG) in human plasma after a nasal dose of 10.4 mg sodium cromoglycate nasal spray, using pravastatin sodium as the internal standard. The method was validated over a linear range of 0.300-20.0 ng/mL. SCG and I.S. were extracted from 1.0 mL of heparinized plasma by C(18) solid-phase extraction cartridges using methanol as eluting solvent. The dried residue was reconstituted with 100 microL of mobile phase, and 10 microL was injected onto the LC-MS/MS system. Chromatographic separation was achieved on a C(18) column (250 x 4.6 mm i.d., 5 microm particle size) with a mobile phase of methanol-acetonitrile-water (containing 2 mmol/L ammonium acetate; 42.5:42.5:15, v/v/v) at a flow rate of 0.4 mL/min. The analytes were detected with a triple quad LC-MS/MS using ESI with positive ionization. Ions monitored in the multiple reaction monitoring mode were m/z 469.0 (precursor ion) to m/z 245.0 (product ion) for SCG and m/z 447.2 (precursor ion) to m/z327.1 (product ion) for pravastatin sodium (internal standard) The average recovery of SCG from human plasma was 94.88% and the lower limit of quantitation was 0.3 ng/mL. Results from a 3-day validation study demonstrated excellent precision and accuracy across the calibration range of 0.3 20 ng/mL. The method was successfully applied to the pharmacokinetic study of SCG in healthy Chinese volunteers. PMID- 18506697 TI - Identification of N,N-dimethyltryptamine and beta-carbolines in psychotropic ayahuasca beverage. AB - Recently many people have shown great interest in traditional indigenous practices and popular medicine, involving the ingestion of natural psychotropic drugs. We received a request to analyze and determine the nature of a dark green liquid with a dark brown plant sediment, which the police had seized at an airport and inside the home of a person belonging to the 'Santo Daime' religious movement. Gas chromatography/mass spectrometry analysis of the extract identified N,N-dimethyltryptamine, a potent hallucinogen, and the beta-carboline alkaloids harmine and harmaline, revealing monoamine oxidase A-inhibiting properties. These substances are typical components of Ayahuasca, a South American psychotropic beverage obtained by boiling the bark of the liana Banisteriopsis caapi together with the leaves of various admixture plants, principally Psychotria viridis. PMID- 18506698 TI - An environmental scanning electron microscopy study of aqueous gibbsite suspensions. AB - Synthetic gibbsite has been used as a model system for study in the environmental scanning electron microscopy (ESEM), to probe its utility as a tool to study clay dispersions under different conditions of aggregation. We have been able to show that we can study the nature of the platelet interactions as the pH is altered, by imaging the dispersions after water evaporation from the surface to permit the surface of the platelets to be clearly seen. It has been possible to show that at alkaline pH there are very few face-edge contacts between the platelets, consistent with what is known about the charges at high pH on the faces and edges of the gibbsite. In contrast, at lower pHs, when faces and edges have opposite sign charges, there are significantly more platelets touching with edge and faces in contact. Finally, when the salt lithium chloride is added to a dispersion at approximately neutral pH, the plates appear to stack suggesting face-face interactions in the dispersion. Thus, ESEM has been able to demonstrate the variability of packing in gibbsite dispersions and to correlate the structures observed with the known charge distribution on the gibbsite platelets. PMID- 18506700 TI - Abstracts of the Thirty-First Annual Meeting of the American Society of Primatologists. June 18-21, 2008. West Palm Beach, Florida, USA. PMID- 18506701 TI - Intravenous iron: a useful therapeutic tool but not a panacea. PMID- 18506702 TI - Congenital factor X deficiencies with a defect only or predominantly in the extrinsic or in the intrinsic system: a critical evaluation. AB - Congenital Factor X deficiency is commonly classified as type I, in which there is a concomitant decrease of activity and antigen (CRM negative), and in type II, in which activity is low but antigen is normal or near normal (CRM positive). During the past decades it was shown that type II was by itself very heterogeneous. It was shown in fact that some forms showed a defect in all three assay systems (extrinsic, intrinsic, and RVV dependent), whereas others showed a defect only in two of the three systems. Molecular biology analysis, whenever available, has failed so far to supply clear explanations for these discrepancies. The purpose of the present article was an attempt to correlate the clotting activities seen in these two defects with other clotting, chromogenic, immunological assays, and molecular biology results. There are in the literature 10 families that show a predominant defect in the extrinsic system, and four families that show a predominant defect in the intrinsic system. All patients showed a normal, near normal, or reduced level of antigen that is always definitively higher than the clotting counterpart. Molecular biology studies revealed mutations in different exons, namely 2, 4, 5, 6, and 8. These mutations in different exons do not allow any clear genotype-phenotype conclusions, but indicate that mutations in different exons may give rise to the same phenotype. The study underlines the importance of a multipronged evaluation of all cases with Factor X deficiency. In fact only by this approach can an acceptable classification of the defect be reached. PMID- 18506703 TI - Elevated urinary leukotriene E4 levels are associated with hospitalization for pain in children with sickle cell disease. AB - Cysteinyl leukotrienes (CsyLTs) are inflammatory mediators produced by white blood cells. Leukotriene LTE(4) is the stable metabolite of CsyLTs, which can be measured in urine. We tested two hypotheses among children with sickle cell disease (SCD): (1) baseline urinary LTE(4) levels are elevated in children with SCD when compared with controls; and (2) baseline LTE(4) levels are associated with an increased incidence rate of hospitalization for SCD-related pain. Baseline LTE(4) levels were measured in children with SCD (cases) and children without SCD matched for age and ethnicity (controls). Medical records of cases were reviewed to assess the frequency of hospitalization for pain within 3 years of study entry. LTE(4) levels were obtained in 71 cases and 22 controls. LTE(4) levels were higher in cases compared with controls (median LTE(4): 100 vs. 57 pg/mg creatinine, P < 0.001). After adjustment for age and asthma diagnosis, a greater incidence rate of hospitalization for pain was observed among children with SCD in the highest LTE(4) tertile when compared with the lowest (114 vs. 52 episodes per 100 patient-years, P = 0.038). LTE(4) levels are elevated in children with SCD when compared with controls. LTE(4) levels are associated with an increased rate of hospitalizations for pain. PMID- 18506704 TI - A myeloablative conditioning regimen for patients with impaired cardiac function undergoing allogeneic stem cell transplantation: reduced cyclophosphamide combined with etoposide and total body irradiation. AB - To circumvent the cardiac toxicity of high-dose cyclophosphamide (CY) in the myeloablative conditioning for those with cardiac comorbidity, we developed a new cardiac sparing conditioning regimen (VP/rCY/TBI) composed of 12 Gy of total body irradiation (TBI), etoposide (VP-16) (40 mg/kg), and reduced CY (40 mg/kg). We assessed the feasibility of this regimen by retrospectively comparing the outcome of VP/rCY/TBI recipients (n = 18) with that of CY/TBI recipients (n = 140). VP/rCY/TBI recipients had significantly higher cumulative dose of anthracyclines, lower ejection fraction (EF), and poorer Karnofsky performance scales (KPS) than CY/TBI recipients. The cumulative incidences of disease progression were 34.9% in VP/rCY/TBI recipients and 19.0% in CY/TBI recipients (P = 0.33). Despite poorer KPS and more cardiac comorbidity in the VP/rCY/TBI recipients, no difference in the nonprogression mortality rates was observed among recipients of the two regimens (17.5 and 14.3%, respectively, P = 0.96). Severe cardiac toxicity within 28 days after transplantation occurred in 5.9 and 3.6% of VP/rCY/TBI and CY/TBI recipients, respectively (P = 0.64). Graft rejection was not observed in VP/rCY/TBI recipients. There is a possibility that VP/rCY/TBI regimen can be safely administered for patients with pretransplantation cardiac comorbidity while preserving antineoplastic effects. These observations merit further prospective study. PMID- 18506705 TI - High frequency of copy-neutral LOH in MUTYH-associated polyposis carcinomas. AB - Genetic instability is known to drive colorectal carcinogenesis. Generally, a distinction is made between two types of genetic instability: chromosomal instability (CIN) and microsatellite instability (MIN or MSI). Most CIN tumours are aneuploid, whereas MSI tumours are considered near-diploid. However, for MUTYH-associated polyposis (MAP) the genetic instability involved in the carcinogenesis remains unclear, as near-diploid adenomas, aneuploid adenomas and near-diploid carcinomas have been reported. Remarkably, our analysis of 26 MAP carcinomas, using SNP arrays and flow sorting, showed that these tumours are often near-diploid (52%) and mainly contain chromosomal regions of copy-neutral loss of heterozygosity (LOH) (71%). This is in contrast to sporadic colon cancer, where physical loss is the main characteristic. The percentage of chromosomal gains (24%) is comparable to sporadic colorectal cancers with CIN. Furthermore, we verified our scoring of copy-neutral LOH versus physical loss in MAP carcinomas by two methods: fluorescence in situ hybridization, and LOH analysis using polymorphic markers on carcinoma fractions purified by flow sorting. The results presented in this study suggest that copy-neutral LOH is an important mechanism in the tumorigenesis of MAP. PMID- 18506707 TI - Methionine sulfoxide reductase: a novel schizophrenia candidate gene. AB - Methionine sulfoxide reductase (MSRA) is an antioxidant enzyme implicated in protection against oxidative stress and protein maintenance. We have previously reported the association of marker D8S542, located within the MSRA gene, with schizophrenia in the Central Valley of Costa Rica (CVCR). By performing fine mapping analysis, we have now identified a potential three-marker at risk haplotype within MSRA in the same CVCR sample, with a global P-value slightly above nominal significance (P = 0.0526). By sequencing the MSRA gene in individuals carrying this haplotype, we identified a novel 4-base pair deletion 1,792 bases upstream of the MSRA transcription start site. This deletion was significantly under-transmitted to schizophrenia patients in the CVCR sample (P = 0.0292) using FBAT, and this was replicated in a large independent sample of 321 schizophrenia families from the Hispanic population (P = 0.0367). These findings suggest a protective effect of the deletion against schizophrenia. Further, MSRA mRNA levels were significantly lower in lymphoblastoid cell lines of individuals homozygous for the deletion compared to carriers of the normal allele (P = 0.0135), although significance was only evident when genotypes were collapsed. This suggests that the deleted sequence may play a role in regulating MSRA expression. In conclusion, this work points towards MSRA as a novel schizophrenia candidate gene. Further studies into the mechanisms by which MSRA is involved in schizophrenia pathophysiology may shed light into the biological underpinnings of this disorder. PMID- 18506708 TI - Inflammation-induced leukocyte accumulation in injured skeletal muscle: role of mast cells. AB - Inflammation consequent to muscle damage is characterized by an accumulation of leukocytes. Our aim in this study was to determine whether mast cells can modulate inflammation-induced leukocyte trafficking. One approach consisted of giving rats a mast cell-degranulating agent, CMP 48/80, prior to a protocol of lengthening contractions inducing inflammation without neutrophil accumulation; in parallel, other rats were given the mast cell-stabilizing agent, cromolyn, prior to injecting muscle with bupivacaine, which induces neutrophil accumulation. Damage was evaluated through measurement of contractile force and inflammation using histochemical and immunohistochemical methods. Stimulation with CMP 48/80 increased the proportion of degranulated mast cells significantly and neutrophil accumulation occurred with lengthening contractions. With bupivacaine, accumulation of neutrophils decreased by 70% when degranulation was inhibited. These results indicate that mast cells are important in the process governing leukocyte trafficking in skeletal muscle trauma and that targeting their inhibition could be an attractive alternative for control of inflammation. PMID- 18506706 TI - The tryptophan hydroxylase-1 A218C polymorphism is associated with diagnosis, but not suicidal behavior, in borderline personality disorder. AB - While there is some preliminary evidence that the tryptophan hydroxylase I (TPH1) polymorphisms are related to Borderline Personality Disorder (BPD), it is not clear if this association is due to the high rates of suicidal behavior in this patient group. Because of the reported association between TPH1 polymorphisms and suicidal behavior, determining whether TPH1 is related to BPD independent of suicidal behavior is of particular importance. One hundred patients diagnosed with BPD and 101 healthy controls were genotyped for TPH1 intron 7 A218C polymorphism and assessed for impulsiveness and hostility. The BPD patient group had a higher frequency of A allele carriers (AA/AC genotypes) than the control group (chi(2) = 6.12, df = 1, P = 0.01), and differed by genotype frequencies (P = 0.03). Suicide attempter status in the patient group was not related to genotype. Logistic regression analysis controlling for age and gender predicted BPD diagnosis from TPH1 allele group (AA/AC vs. CC, P = 0.03), and TPH1 heterozygotes (AC) appeared to have the highest risk for BPD (P = 0.03). In the full sample, participants with the AC genotype had higher impulsiveness and hostility scores. However, TPH1 did not predict these traits in either of the groups independently, suggesting the association may be an artifact of the association between TPH1 and BPD. Results suggest that the A allele of the tryptophan hydroxylase-1 A218 polymorphism may be associated with BPD, and that it does not appear to be related to suicidal behavior in this population. An aspect of BPD pathology may be due to serotonergic dysfunction. PMID- 18506709 TI - Bilateral drop foot secondary to a primary tumor in the conus medullaris. AB - Intramedullary tumors of the spinal cord are rare neoplasms that can be associated with severe neurological and functional handicaps. To our knowledge, we describe for the first time the case of a male patient who developed bilateral drop foot as an initial manifestation of a primary tumor in the conus medullaris of the spinal cord, probably an astrocytoma. PMID- 18506710 TI - Changes in Na(+) channel expression and nodal persistent Na(+) currents associated with peripheral nerve regeneration in mice. AB - Patients with peripheral neuropathy frequently suffer from positive sensory (pain and paresthesias) and motor (muscle cramping) symptoms even in the recovery phase of the disease. To investigate the pathophysiology of increased axonal excitability in peripheral nerve regeneration, we assessed the temporal and spatial expression of voltage-gated Na(+) channels as well as nodal persistent Na(+) currents in a mouse model of Wallerian degeneration. Crushed sciatic nerves of 8-week-old C57/BL6J male mice underwent complete Wallerian degeneration at 1 week. Two weeks after crush, there was a prominent increase in the number of Na(+) channel clusters per unit area, and binary or broad Na(+) channel clusters were frequently found. Excess Na(+) channel clusters were retained up to 20 weeks post-injury. Excitability testing using latent addition suggested that nodal persistent Na(+) currents markedly increased beginning at week 3, and remained through week 10. These results suggest that axonal regeneration is associated with persistently increased axonal excitability resulting from increases in the number and conductance of Na(+) channels. PMID- 18506712 TI - Muscle ultrasound in neuromuscular disorders. AB - Muscle ultrasound is a useful tool in the diagnosis of neuromuscular disorders, as these disorders result in muscle atrophy and intramuscular fibrosis and fatty infiltration, which can be visualized with ultrasound. Several prospective studies have reported high sensitivities and specificities in the detection of neuromuscular disorders. Although not investigated in large series of patients, different neuromuscular disorders tend to show specific changes on muscle ultrasound, which can be helpful in differential diagnosis. For example, Duchenne muscular dystrophy results in a severe, homogeneous increase of muscle echo intensity with normal muscle thickness, whereas spinal muscular atrophy shows an inhomogeneous increase of echo intensity with severe atrophy. A major advantage of muscle ultrasound, compared to other imaging techniques, is its ability to visualize muscle movements, such as muscle contractions and fasciculations. This study reviews the possibilities and limitations of ultrasound in muscle imaging and its value as a diagnostic tool in neuromuscular disorders. PMID- 18506711 TI - A novel SNaPshot assay to detect the mdx mutation. AB - The mdx mouse is an animal model for Duchenne muscular dystrophy (DMD). In order to evaluate possible treatments and to carry out genetic studies, it is essential to distinguish between mice that carry the dystrophic (mutant) or wild-type (wt) allele(s). The current amplification-resistant mutation system (ARMS) assay is labor intensive and yields false negatives, which reduces its efficiency as a screening tool. An alternate assay based on single-nucleotide polymorphism (SNP) primer extension technology (i.e., SNaPshot) is described. The SNaPshot assay has been optimized to identify both wild-type and mutant alleles, providing a robust, potentially automatable assay for high-throughput analysis. PMID- 18506713 TI - Amyloidogenic transthyretin Val30Met homozygote showing unusually early-onset familial amyloid polyneuropathy. AB - We report an amyloidogenic transthyretin (ATTR) Val30Met homozygote showing extremely early-onset, severe familial amyloid polyneuropathy (FAP). Although homozygotes have been reported to show late-onset and mild clinical manifestations, detailed analyses of the present and previously reported families suggest that homozygotes have a slightly more severe clinical course than heterozygotes. This is the youngest reported patient with ATTR Val30Met FAP, a condition believed to be attributable to homozygosity of this mutation. The clinical severity is consistent with TTR protein instability. PMID- 18506714 TI - Effects of changes in intracellular action potential on potentials recorded by single-fiber, macro, and belly-tendon electrodes. AB - Some myopathies are accompanied by abnormal calcium homeostasis. Electromyography (EMG) in such patients shows signs of normal or myopathic EMG when detected by a single-fiber electrode and abnormally increased values in macro EMG. As calcium accumulation might be accompanied by changes in intracellular action potential (IAP) and muscle-fiber propagation velocity, we simulated the effects of such changes on motor unit potentials (MUPs) recorded by different kinds of electrodes. We found that: (1) the requirements for what potential can be accepted as a single-fiber action potential (SFAP) are too rigorous; (2) macro MUP amplitude can increase while SFAP amplitude can decrease when there is an increase in the spatial length of IAP spike; and (3) changes in the second phase of a belly-tendon-detected MUP or M wave could be used for noninvasive detection of increased IAP depolarizing (negative) after-potential. PMID- 18506716 TI - Changes in intracellular action potential profile affect parameters used in turns/amplitude analysis. AB - The influence of changes in the intracellular action potential (IAP) spatial profile on motor unit potentials (MUPs), number of turns per second (NTs), and mean turn amplitude were simulated and analyzed. We show why measurement of NTs was "the best indicator of neurogenic affection" and why the lower diagnostic yield of turns/amplitude analysis in myopathy could be due to changes in IAP shape caused by elevated free calcium concentration. The results explain the complications observed when interference electromyographic signals obtained during high levels of isometric contractions were analyzed. We show that, in contrast to earlier assumptions, the effect of increased IAP spike duration on NTs was stronger than that of a decrease in muscle fiber propagation velocity (MFPV). The decrease in the NTs could occur without a drop-out of MUs and/or a decrease in their firing rates, and without a change in MFPV and synchronous firing. PMID- 18506718 TI - Stop using arbitrary grading schemes in carpal tunnel syndrome. PMID- 18506719 TI - Childhood multifocal acquired demyelinating sensory and motor neuropathy. AB - We report the first pediatric cases of multifocal acquired demyelinating sensory and motor neuropathy with electrophysiologic evidence of proximal conduction abnormalities but no definite conduction block. Intravenous immunoglobulin caused clinical improvement followed by long-term remission without maintenance therapy; one patient has exhibited a monophasic course and the other has had a single relapse during the last 5 years. These cases suggest that there may be a long term sustained beneficial effect of intravenous immunoglobulin therapy for children with this neuropathy. PMID- 18506720 TI - Reference jitter values for concentric needle electrodes in voluntarily activated extensor digitorum communis and orbicularis oculi muscles. AB - The aim of this study was to estimate normal jitter in voluntarily activated extensor digitorum communis (EDC) and orbicularis oculi (OOc) muscles using a disposable concentric needle electrode (CNE). The EDC of 67 normal subjects (22 males and 45 females, mean age 35.5 +/- 10.2 years) and the OOc of 50 normal subjects (13 males and 37 females, mean age 37.9 +/- 9.6 years) were studied. Jitter values were expressed as the mean consecutive difference (MCD) of 20 potential pairs. The mean MCD for EDC was 23.6 +/- 3.1 micros (upper 95% confidence limit [CL]: 29.7 micros). The mean MCD of all potential pairs (n = 1340) was 23.5 +/- 7.3 micros (95% CL: 38.2 micros). The mean MCD for the 18th highest value was 31.4 +/- 4.9 micros (95% CL: 41.2 micros). The mean MCD for OOc was 24.7 +/- 3.1 micros (95% CL: 31.0 micros). The mean MCD of all potential pairs (n = 1000) was 24.7 +/- 7.1 micros (95% CL: 39.0 micros). The mean MCD for the 18th highest value was 32.7 +/- 4.1 micros (95% CL: 40.9 micros). Our reported CNE jitter values obtained during voluntary activation represent the largest series currently available. The suggested practical limit in the EDC for mean MCD was 30 mus and for outliers was 42 micros, and in the OOc for mean MCD was 31 micros and 41 micros for outliers. The present study confirms that CNE can be used to assess jitter values, although certain precautions must be taken. PMID- 18506721 TI - A new method for measuring compound muscle action potentials in facial palsy: a preliminary study. AB - To establish a simple, reproducible procedure for studying facial motor nerve conduction (MNC), we determined the optimal electrode position to record evoked compound muscle action potentials (CMAPs) from perioral muscles in normal subjects. We examined three new electrode positions in which the electrode connected to the one input of the amplifier was placed on the mental protuberance, and the one connected to the other input was placed on the skin over the orbicularis oris muscle (the philtrum, mouth angle, or lower lip). We then compared the morphology and amplitudes of the CMAPs, right-left differences, and the reproducibility of CMAP amplitudes with recordings taken from the standard electrode position in which one electrode was placed on the nasolabial fold closely lateral to the ala nasi, and the other was placed on the skin over the orbicularis oris. Percutaneous supramaximal electrical stimulation was applied to the main trunk of the facial nerve. All three of the new recording positions showed greater amplitudes and more obvious biphasic CMAPs than the standard method. Positioning the electrode connected to the negative input on the philtrum was optimal in terms of right-left differences and the reproducibility of CMAP amplitudes. Therefore, this midline recording is a simple, reproducible method for calculating the CMAP amplitude ratio. However, prior to clinical use of this procedure, analyses of patients with facial palsy are required. PMID- 18506722 TI - Lambert-Eaton syndrome with large-cell neuroendocrine carcinoma of the lung. AB - Lambert-Eaton syndrome (LES) is an immune-mediated disorder of the presynaptic neuromuscular junction due to the blocking effect of the voltage-gated calcium channel (VGCC) antibodies. Small-cell lung cancer (SCLC) is the most common cause of LES. We report an unusual case of LES associated with large-cell neuroendocrine carcinoma (LCNEC) of the lung. In this case, clinical symptoms of LES predated the diagnosis of LCNEC by 6 years. After tumor resection, the patient experienced clinical and electrophysiological improvement. In addition, he had a decrease in VGCC antibody titer from 130 to 80 pmol/L. The onset of LES can be prolonged, and tumor surveillance should continue in these cases. PMID- 18506723 TI - Re: Comparison of perception threshold testing and thermal-vibratory testing, MUS. 2008 Apr; 37(4): 514-7. PMID- 18506725 TI - Identification of major xanthones and steroidal saponins in rat urine by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technology following oral administration of Rhizoma Anemarrhenae decoction. AB - Rhizoma Anemarrhenae (Zhimu in Chinese), the dried rhizome of Anemarrhena asphodeloides Bge. (Fam. Liliaceae), is a well-known traditional Chinese medicinal herb and has been used clinically in China for centuries to cure various diseases. However, like other traditional Chinese medicines, the effective constituents of this medicine, especially the assimilation and metabolites in vivo, which are very important to show their effects, have not been systematically studied. In this paper, solid-phase extraction and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technologies were used to study the constituents absorbed into rat urine and their metabolites after oral administration of Rhizoma Anemarrhenae decoction. A total of 11 compounds, including two xanthones, three of their metabolites and six steroidal saponins, were identified in rat urine sample. They were neomangiferin (1), glucuronide and monomethyl conjugate of mangiferin (2), mangiferin (3), monomethyl conjugate of mangiferin (4), dimethyl conjugate of mangiferin (5), timosaponin N or timosaponin E1 (6), timosaponin BII (7), timosaponin BIII (8), anemarrhenasaponin I or anemarrhenasaponin II (9), timosaponin AII (10) and timosaponin AIII (11). The results would efficaciously narrow the potentially active compounds range in Rhizoma Anemarrhenae decoction, and pave a helpful way for follow-up mechanism of action research. PMID- 18506726 TI - A science-based approach to managing disease risks for ape conservation. PMID- 18506727 TI - Separation assay of histamine and its metabolites in biological specimens. AB - This review summarizes the determination methods for histamine and the metabolites in biological specimens by separation techniques, such as gas chromatography, liquid chromatography and capillary electrophoresis. The typical applications using these methods are also described in this review together with the characteristics of the methods. PMID- 18506728 TI - Simultaneous determination of seven bioactive lignans in Herpetospermum caudigerum by RP-HPLC method. AB - A reversed-phase high-performance liquid chromatographic method with diode array detection was established to simultaneously determine the seven bioactive lignans in Herpetospermum caudigerum, namely ent-isolariciresinol (1), dehydrodiconiferyl alcohol (2), herpetrione (3), herpetin (4), herpetetrone (5), herpetotriol (6) amd herpetal (7). The HPLC assay was performed on a Restek Pinnacle DB C18 column (250 x 4.6 mm, 5 microm) with gradient elution of acetonitrile and 0.1% phosphoric acid within 65 min. The detection wavelength was 240 nm. The flow-rate was 1.0 mL/min. All calibration curves showed good linearity (r2 > 0.9998) within test ranges. The method was reproducible with intra- and inter-day variation of less than 1.98%. The method provided good accuracy with recoveries in the range 95.19-102.64% with RSDs less than 1.52%. The method was successfully applied to the quantification of seven constituents in 15 H. caudigerum samples collected from different cities. The results indicated that the developed assay could be considered as a suitable quality control method for H. caudigerum. PMID- 18506729 TI - Identification of differentially expressed proteins in the heart of translationally controlled tumor protein over-expressing transgenic mice. AB - We previously reported that transgenic (TG) mice over-expressing translationally controlled tumor protein (TCTP) developed systemic arterial hypertension at about 6 weeks after birth. In the present study, we identified, using proteomics technologies, 24 other proteins that were differentially expressed in the heart of TCTP over-expressing TG mice. These 24 proteins are involved in a variety of biological processes such as reactive oxygen species metabolism, cytoskeleton organization, fatty acid metabolism, amino acid metabolism and energy metabolism. We determined protein expression levels of the peroxiredoxin (Prx)2, Prx3, myosin light chain 1, stress protein (heat shock protein) 25K, and T-complex protein 1 alpha subunit by western blot analysis. Over-expression of TCTP probably regulates the expression of other proteins which play a pivotal role in a variety of cellular functions in TCTP over-expressing TG mice. PMID- 18506730 TI - Analysis of iridoids from Harpagophytum and eleutherosides from Eleutherococcus senticosus in horse urine. AB - LC/ESI-MS n methods have been previously set up to detect the administration of (i) Harpagophytum and (ii) preparations containing a plant capable of anti-stress properties: Eleutherococcus senticosus. Harpagoside has been found to be the main indicator of Harpagophytum administration in the horse. These methods have been applied to a large number of horse urine samples of various origins. Regarding the detection of Harpagophytum administration, harpagoside, harpagide and 8-para coumaroyl harpagide were detected together in only one sample out of 317. Eleutheroside E was found to be the main indicator of Eleutherococcus senticosus administration. It was detected in post-administration samples collected from two horses having received a feed supplement containing Eleutherococcus senticosus for several days. Out of the 382 samples tested, eleutheroside E was found in an unexpected large number of urine samples (39%) of various origins and its presence cannot be only due to the sole use of herbal dietary supplements. PMID- 18506731 TI - Analysis of genetic variations in the human Par-4 (PAWR) gene and tardive dyskinesia in schizophrenia. PMID- 18506732 TI - Causes of death in the Kasekela chimpanzees of Gombe National Park, Tanzania. AB - Understanding the rates and causes of mortality in wild chimpanzee populations has important implications for a variety of fields, including wildlife conservation and human evolution. Because chimpanzees are long-lived, accurate mortality data requires very long-term studies. Here, we analyze 47 years of data on the Kasekela community in Gombe National Park. Community size fluctuated between 38 and 60, containing 60 individuals in 2006. From records on 220 chimpanzees and 130 deaths, we found that the most important cause of mortality in the Kasekela community was illness (58% of deaths with known cause), followed by intraspecific aggression (20% of deaths with known cause). Previous studies at other sites also found that illness was the primary cause of mortality and that some epidemic disease could be traced to humans. As at other study sites, most deaths due to illness occurred during epidemics, and the most common category of disease was respiratory. Intraspecific lethal aggression occurred within the community, including the killing of infants by both males and females, and among adult males during the course of dominance-related aggression. Aggression between communities resulted in the deaths of at least five adult males and two adult females in the Kasekela and Kahama communities. The frequency of intercommunity violence appears to vary considerably among sites and over time. Intercommunity lethal aggression involving the Kasekela community was observed most frequently during two periods. Other less common causes of death included injury, loss of mother, maternal disability, and poaching. PMID- 18506733 TI - Mortality after cure of soft-tissue sarcoma treated with conservation surgery and radiotherapy. AB - BACKGROUND: The objective of the current study was to analyze the potential treatment-related mortality in long-term survivors of soft-tissue sarcoma (STS) treated with radiotherapy (RT) and conservation surgery. METHODS: Between 1960 and 2000, 629 of 1089 patients treated with conservation surgery and RT for nonmetastatic STS at the University of Texas M. D. Anderson Cancer Center never developed disease recurrence. Long-term survival and causes of death were evaluated using the person-years method to determine the standardized mortality ratio (SMR). SMRs were calculated for death from all causes, cancer, and cardiac disease using standard U.S. data. RESULTS: The median follow-up was 13.2 years. The 10-year, 20-year, and 30-year actuarial survival rates were 88%, 69%, and 52%, respectively. The overall all-case mortality was 1.14 (95% confidence interval [95% CI], 0.98-1.33). The all-cause mortality exceeded that expected for female patients with an SMR of 1.48 (95% CI, 1.15-1.88), patients aged 263.8, and for IS was m/z 349.0 --> 205.9. The linearity was established over the concentration range of 1.56-400.00 ng/mL. The inter-day and the intra-day precisions were all within 10%. A simple protein precipitation with methanol was adopted for sample preparation. The extraction recoveries of ambroxol and IS were higher than 90.80%. The validated method was successfully applied in pharmacokinetic study after oral administration of 90 mg ambroxol to 24 healthy volunteers. PMID- 18506739 TI - Microwave-assisted micellar extraction coupled with solid-phase extraction for preconcentration of pharmaceuticals in molluscs prior to determination by HPLC. AB - A simple and specific analytical method was developed and tested for the determination of pharmaceuticals in mollusc samples. A combination of microwave assisted micellar extraction (MAME) and solid-phase extraction (SPE) using a non ionic surfactant, polyoxyethylene 10 lauryl ether, was examined to extract and determine simultaneously a group of pharmaceuticals such as carbamazepine, clorfibric acid, ketoprofen, naproxen, bezafibrate and ibuprofen by liquid chromatography using UV-diode array detector. The MAME extraction performance was evaluated by studying various parameters such as the volume and concentration of surfactant and microwave conditions. Finally, an OASIS HLB cartridge was used as an optimum SPE sorbent to clean up the extracts and preconcentrate the selected analytes. The proposed method showed satisfactory linearity and reproducibility (between 3 and 15%), as well as detection limits ranging from 30 to 220 ng/g. Finally, the method was successfully applied to the determination of the target pharmaceuticals in various kinds of mollusc samples. This study has demonstrated that microwave-assisted micellar extraction with solid-phase extraction may be used as a viable alternative to conventional methods for the extraction of pharmaceuticals in this type of matrices. PMID- 18506740 TI - A high-performance liquid chromatographic method for determination of scopolin in rat plasma: application to pharmacokinetic studies. AB - An analytical method based on high-performance liquid chromatographic (HPLC) with ultraviolet (UV) detection was developed for determination of scopolin in rat plasma using aesculin as internal standard (IS). After protein precipitation of plasma sample with methanol, the supernatant was directly injected and analyzed. Chromatographic separation was achieved on a C18 column using methanol and distilled water (22:78, v/v) containing 0.2% (v/v) glacial acetic acid as mobile phase with a column temperature of 30 degrees C. The UV detector was set at 338 nm. The calibration curve was linear over the range of 0.105-13.125 microg/mL with a correlation coefficient of 0.9998. The retention times of aesculin and scopolin were 10.4 and 12.8 min, respectively. The recoveries for plasma samples of 0.105, 4.725 and 13.125 microg/mL were 91.08, 95.30 and 96.10%, respectively. The RSD of intra- and inter-day assay variations was less than 7.35%. The lower limit of detection was 0.03 microg/mL .This HPLC assay is a simple, sensitive and accurate and was successfully applied to the pharmacokinetic study of scopolin in rats. PMID- 18506741 TI - Ultra-fast chromatographic micro-assay for quantification of diphenidol in plasma: application in an oral multi-dose switchability trial. AB - Pharmacokinetics of diphenidol (DPN) is limited due to the lack of analytical methodology. Here, a micro-assay for DPN quantification was developed, by coupling ultra-performance liquid chromatography with tandem mass spectrometry. The procedure involved plasma precipitation and injection of supernatant into UPLC with an Acquitytrade mark C18 column. Detection was in positive electrospray, following transitions of m/z 310.3 --> 292.3 and m/z 275.3 --> 230.2 for DPN and chlorphenamine (internal standard), respectively. The method was linear with a range of 4-400 ng/mL, and a 2 min run time. This method was applied in a switchability trial, where both formulations of DPN were bioequivalent. PMID- 18506742 TI - Determination of organic acids in urine by solid-phase microextraction and gas chromatography-ion trap tandem mass spectrometry previous 'in sample' derivatization with trimethyloxonium tetrafluoroborate. AB - A method for the determination of the organic acids directly in the urine employing derivatization with trimethyloxonium tetrafluoroborate as a methylating agent and sequential extraction by head space and direct immersion/solid phase microextraction is reported. Furoic acid, hippuric acid, methylhippuric acid, mandelic acid, phenylglyoxylic acid and trans, trans muconic acid contained in urine and proposed by the American Conference of Governmental Industrial Hygienists as biological exposure indices were determined after a fast and economically convenient preparation step and sensitive gas chromatography-ion trap-mass spectrometry/tandem mass spectrometry analysis. Urine is rather a complex sample and hence the acquisition method required specific GC-MS instrumentation capable of supporting the changeover, fully automated during a single chromatographic separation, from mass to tandem mass spectrometry and both chemical and electron ionization modes. The automation of the analytical method provides a number of advantages, including reduced analysis time for both routine analysis and method development, and greater reproducibility. The equilibrium and kinetics of this substances vs head space/direct immersion-solid phase microextraction were investigated and evaluated theoretically. PMID- 18506743 TI - Screening and mechanism study of components targeting DNA from the Chinese herb Lonicera japonica by liquid chromatography/mass spectrometry and fluorescence spectroscopy. AB - A method which involves combination of centrifugal ultrafiltration sampling with high-performance liquid chromatography coupled with diode array detection and mass spectrometry (HPLC-DAD-MS) analysis was established for screening bioactive compounds binding to calf thymus deoxyribonucleic acid (ct-DNA) from the extracts of Lonicera japonica. Four compounds were screened out and identified as rutin, quercetin-3-O-glucoside, luteolin-7-O-glucoside and lonicerin, based on the comparison of retention time, UV spectra and MS data with those of standards. The DNA-binding capabilities of the latter three flavonoids were found for the first time. The binding mechanisms of rutin, quercetin-3-O-glucoside and luteolin-7-O glucoside with ct-DNA at the molecular level were explored using acridine orange (AO) as a fluorescence probe. Groove binding is the most appropriate binding mode of these three flavonoids to DNA, according to ultraviolet absorption and fluorescence spectra, as well as melting temperature (T(m)) curves and viscosity measurements. The binding constants of rutin, quercetin-3-O-glucoside and luteolin-7-O-glucoside with DNA-AO complex were 3.81 x 10(3), 3.37 x 10(3) and 5.50 x 10(3) L/mol, respectively. PMID- 18506744 TI - Glycosaminoglycan in cerebrum, cerebellum and brainstem of young sheep brain with particular reference to compositional and structural variations of chondroitin dermatan sulfate and hyaluronan. AB - Recent advances in the structural biology of chondroitin sulfate chains have suggested important biological functions in the development of the brain. Several studies have demonstrated that the composition of chondroitin sulfate chains changes with aging and normal brain maturation. In this study, we determined the concentration of all glycosaminoglycan types, i.e. chondroitin sulfate, dermatan sulfate, keratan sulfate, heparan sulfate, hyaluronan and chondroitin in cerebrum, cerebellum and brainstem of young sheep brain. In all cases, chondroitin sulfate was the predominant glycosaminoglycan type, comprising about 54-58% of total glycosaminoglycans, with hyaluronan being present also in significant amounts of about 19-28%. Of particular interest was the increased presence of the disulfated disaccharides and dermatan sulfate in cerebellum and brainstem, respectively, as well as the detectable and measurable occurrence of chondroitin in young sheep brain. Among the three brain areas, cerebrum was found to be significantly richer in chondroitin sulfate and hyaluronan, two major extracellular matrix components. These findings imply that the extracellular matrix of the cerebrum is different from those of cerebellum and brainstem, and probably this fact is related to the particular histological and functional characteristics of each anatomic area of the brain. PMID- 18506745 TI - Lipoma of the right atrium. AB - A 66-year-old asymptomatic woman was admitted to our hospital with the diagnosis of a right atrial mass detected on an outside transthoracic echocardiogram and confirmed on transesophageal echocardiography. Physical examination and basal electrocardiogram were normal. Transthoracic echocardiography revealed a 3.8 x 2.5 cm echogenic mass in the right atrium. A multislice CT examination demonstrated a right atrial mass with a fat density ranging from -80 to -110 HU. The patient had a successful surgical excision of the mass, and the diagnosis of lipoma was confirmed on histopathological examination. PMID- 18506746 TI - Preoperative sonography of nonreducible inguinal masses in girls. AB - PURPOSE: Inguinal hernia is one of the most common surgical pathologies in childhood. Any of the abdominal organs can slide into the hernial sac and become incarcerated there. In girls, the fallopian tubes, ovaries, uterus, and-rarely ovarian cysts can form the sliding component of an inguinal hernia. The aim of this study was to investigate the diagnostic value of preoperative sonographic examination in girls with nonreducible inguinal masses. METHODS: Nine girls ranging in age from 2 months to 8 years who were admitted to our clinic with nonreducible inguinal masses were included in the study. All patients underwent sonographic examination followed by surgery on the day of admission. RESULTS: A definitive diagnosis was obtained in 6 patients on preoperative sonographic evaluation, whereas 3 patients were misdiagnosed. One patient was diagnosed sonographically as having lymphadenopathy, but surgery revealed an ovarian cyst sliding into the hernial sac. A second patient was found to have an infected lymph node at surgery instead of a strangulated bowel loop as diagnosed on sonographic examination. In the third patient, the preoperative sonographic diagnosis was an ovarian cyst in the hernia sac, but surgery revealed a cyst of the canal of Nuck. CONCLUSION: Inguinal masses in young girls must be carefully evaluated, because the sonographic preoperative diagnosis may be misleading. PMID- 18506747 TI - Prenatal diagnosis of trisomy 16 mosaicism manifested as pulmonary artery stenosis. AB - Trisomy 16 mosaicism detected at midtrimester amniocentesis is rare and indicative of true fetal mosaicism. We report a case of mosaic trisomy 16 diagnosed by amniocentesis in which the sonographic findings included fetal pulmonary artery stenosis, a single umbilical artery, and early onset fetal growth restriction. The pregnancy was legally terminated. A review of previous reports suggests that abnormalities of outlet tracts are rarely encountered in fetuses with trisomy 16 mosaicism revealed via amniocentesis. PMID- 18506748 TI - Identification of copy number gain and overexpressed genes on chromosome arm 20q by an integrative genomic approach in cervical cancer: potential role in progression. AB - Recurrent karyotypic abnormalities are a characteristic feature of cervical cancer (CC) cells, which may result in deregulated expression of important genes that contribute to tumor initiation and progression. To examine the role of gain of the long arm of chromosome 20 (20q), one of the common chromosomal gains in CC, we evaluated CC at various stages of progression using single nucleotide polymorphism (SNP) array, gene expression profiling, and fluorescence in situ hybridization (FISH) analyses. This analysis revealed copy number increase (CNI) of 20q in >50% of invasive CC and identified two focal amplicons at 20q11.2 and 20q13.13 in a subset of tumors. We further demonstrate that the acquisition of 20q gain occurs at an early stage in CC development and the high-grade squamous intraepithelial lesions (HSIL) that exhibit 20q CNI are associated (P = 0.05) with persistence or progression to invasive cancer. We identified a total of 26 overexpressed genes as consequence of 20q gain (N = 14), as targets of amplicon 1 (N = 9; two genes also commonly expressed with 20q gain) and amplicon 2 (N = 6; one gene also commonly expressed with 20q gain). These include a number of functionally important genes in cell cycle regulation (E2F1, TPX2, KIF3B, PIGT, and B4GALT5), nuclear function (CSEL1), viral replication (PSMA7 and LAMA5), methylation and chromatin remodeling (ASXL1, AHCY, and C20orf20), and transcription regulation (TCEA2). Our findings implicate a role for these genes in CC tumorigenesis, represent an important step toward the development of clinically significant biomarkers, and form a framework for testing as molecular therapeutic targets. PMID- 18506749 TI - Novel regions of acquired uniparental disomy discovered in acute myeloid leukemia. AB - The acquisition of uniparental disomy (aUPD) in acute myeloid leukemia (AML) results in homozygosity for known gene mutations. Uncovering novel regions of aUPD has the potential to identify previously unknown mutational targets. We therefore aimed to develop a map of the regions of aUPD in AML. Here, we have analyzed a large set of diagnostic AML samples (n = 454) from young adults (age: 15-55 years) using genotype arrays. Acquired UPD was found in 17% of the samples with a nonrandom distribution particularly affecting chromosome arms 13q, 11p, and 11q. Novel recurrent regions of aUPD were uncovered at 2p, 17p, 2q, 17q, 1p, and Xq. Overall, aUPDs were observed across all cytogenetic risk groups, although samples with aUPD13q (5.4% of samples) belonged exclusively to the intermediate risk group as defined by cytogenetics. All cases with a high FLT3-ITD level, measured previously, had aUPD13q covering the FLT3 gene. Significantly, none of the samples with FLT3-ITD(-)/FLT3-TKD(+) mutation exhibited aUPD13q. Of the 119 aUPDs observed, the majority (87%) were due to mitotic recombination while only 13% were due to nondisjunction. This study demonstrates aUPD is a frequent and significant finding in AML and pinpoints regions that may contain novel mutational targets. PMID- 18506751 TI - Human and software error in ductus venosus Doppler waveform analysis. AB - Ductus venosus waveform analysis has become increasingly affected by technical errors; however, these errors could be avoided if more attention was paid during sampling and analysis. The most common misevaluations include incorrect tracing, under- or overestimation of the peak systolic velocity, overestimation of the end diastolic velocity, and, as a consequence, incorrect calculation of the pulsatility index facilitated either by human or software error. This article proposes practical suggestions to avoid technical errors in ductus venosus waveform analysis. PMID- 18506750 TI - Inactivation of the cystatin E/M tumor suppressor gene in cervical cancer. AB - We have previously localized a cervical cancer tumor suppressor gene to a 300 kb interval of 11q13. Analysis of candidate genes revealed loss of expression of cystatin E/M, a lysosomal cysteine protease inhibitor, in 6 cervical cancer cell lines and 9 of 11 primary cervical tumors. Examination of the three exons in four cervical cancer cell lines, 19 primary tumors, and 21 normal controls revealed homozygous deletion of exon 1 sequences in one tumor. Point mutations were observed in six other tumors. Two tumors contained mutations at the consensus binding sites for cathepsin L, a lysosomal protease overexpressed in cervical cancer. Introduction of these two point mutations using site directed mutagenesis resulted in reduced binding of mutated cystatin E/M to cathepsin L. Although mutations were not observed in any cell lines, four cell lines and 12 of 18 tumors contained promoter hypermethylation. Reexpression of cystatin E/M was observed after 5'aza 2-deoxycytidiene and/or Trichostatin A treatment of cervical cancer cell lines, HeLa and SiHa, confirming promoter hypermethylation. Ectopic expression of cystatin E/M in these two cell lines resulted in growth suppression. There was also suppression of soft agar colony formation by HeLa cells expressing the cystatin E/M gene. Reexpression of cystatin E/M resulted in decreased intracellular and extracellular expression of cathepsin L. Overexpression of cathepsin L resulted in increased cell growth which was inhibited by the reintroduction of cystatin E/M. We conclude, therefore, that cystatin E/M is a cervical cancer suppressor gene and that the gene is inactivated by somatic mutations and promoter hypermethylation. PMID- 18506752 TI - Prenatal diagnosis of Pentalogy of Cantrell in the first trimester: is 3 dimensional sonography needed? AB - We report the prenatal diagnosis of 2 cases of Pentalogy of Cantrell in the first trimester. In case 1, sonographic evaluation revealed ectopia cordis, omphalocele, and cystic hygroma at 10 weeks' gestation. In case 2, sonographic assessment during the first trimester detected ectopia cordis and omphalocele at 11 weeks' gestation. In both cases, the patient opted for elective pregnancy termination, and Pentalogy of Cantrell was confirmed in 2 male fetuses. We discuss the role of Doppler imaging and 3-dimensional sonography as complementary methods to conventional sonographic assessment of abdominal wall defects at early pregnancy. PMID- 18506753 TI - Internet use among adolescent and young adults (AYA) with cancer. AB - BACKGROUND: The Internet serves as an important resource for adult cancer patients, but little is known about Internet use among adolescent and young adults (AYA) with cancer. The aims of this study were to describe (1) cancer specific websites which AYA with cancer visit and (2) Internet features desired by AYA on cancer-specific websites and how many current AYA cancer websites contain these features. PROCEDURE: Individual phone interviews were conducted with a convenience sample of 16 AYA with cancer from across North America in June 2005. Content analysis of these interviews were coded and validated for desired website features. Current AYA cancer websites were identified on the Internet and the features on these sites were compared to the features desired by our sample. RESULTS: Favorite websites visited by AYA with cancer (cancer-related and unrelated) were identified along with current Internet use. Twenty-one distinct cancer website features desired by AYA with cancer were described. Twenty-seven unique AYA cancer websites were found on the Internet during May-June 2006. Each site contained 7.7 (SD = 2.7) of the 21 features identified by participants as desirable, but the highest ranked features did not occur in the majority of these websites. CONCLUSIONS: AYA with cancer indicate that they prefer to visit cancer websites that contain cancer-related information, provide the ability to chat with AYA with cancer, and offer some type of game. Although many websites exist for AYA with cancer, few individual sites contain the web features identified as most desired by AYA with cancer. PMID- 18506754 TI - Severe chronic refractory immune thrombocytopenic purpura during childhood: a survey of physician management. AB - BACKGROUND: Physician attitudes regarding management of children with severe chronic immune thrombocytopenic purpura (ITP) have not been recently characterized. PROCEDURE: We designed a survey of members of the American Society of Pediatric Hematology-Oncology (ASPHO) that described a 5-year-old female with ITP for 1 year who was unresponsive to steroids, IVIG, and anti-D immune globulin and having frequent epistaxis causing interference with her daily activities. A 13-item questionnaire evaluated physician decision-making in this setting. RESULTS: Two hundred and ninety-seven surveys (35% response rate) were returned, and 295 were evaluable. Thirty-three percent of respondents stated that they would recommend splenectomy for such a child. Of those who would not recommend splenectomy, 67% reported that they would instead treat with rituximab. If initial drug therapy failed, 47% would proceed with splenectomy. Those who reported treating with rituximab initially were more likely to recommend splenectomy following failure than those who preferred other drug therapy (P < 0.0001). CONCLUSIONS: Physician management of patients with chronic ITP is diverse. With the advent of new treatments such as rituximab and thrombopoetic agents it is critically important to compare their cost, adverse effects and efficacy with splenectomy in order to optimally guide treatment practices. PMID- 18506755 TI - Hyperferritinemia in hemophagocytic lymphohistiocytosis and related diseases. PMID- 18506756 TI - "When can I go home?"-seeking ways to lower the burden on patients and families. PMID- 18506757 TI - Numb chin syndrome as initial manifestation in a child with acute lymphoblastic leukemia. AB - Numb chin syndrome (NCS) describes the affection of the inferior alveolar nerve and is a purely sensory neuropathy. Its symptoms include numbness of the skin of the chin, the lip and the gingival mucosa. Mostly seen in adults, it has rarely been described in children. We report on an 11-year-old male who presented with NCS as initial manifestation of acute lymphoblastic leukemia of B-cell type. PMID- 18506758 TI - Conservative management of priapism secondary to leukemia. AB - We report four cases of leukemia (three chronic myeloid and one T-cell acute lymphoblastic) presenting with priapism in children 9- to 13-year old. All of them presented with hyperleukocytosis, and three had anemia plus thrombocytosis. All patients underwent chemotherapy and two had leukopheresis. In all cases, priapism was managed conservatively. Erection required up to 13 days to start improving but none of the patients developed clinical evidence of long-term erectile dysfunction. Based on these cases, conservative management of priapism in children with leukemia might be adequate and not lead to long-term erectile dysfunction. PMID- 18506759 TI - Host factors and consequence of chemotherapy in pediatric cancer patients. AB - The 5-year survival rates for childhood ALL are approaching 80%, but therapy related toxicities are common. One of the challenges in this population is determining the most efficacious therapeutic regimens for these individuals. Factors such as drug metabolism, genetics, and concomitant disease affect drug efficacy and may be important in determining therapeutic outcomes in these patients. This review will focus on these factors in the treatment of childhood ALL. PMID- 18506761 TI - The transcription factor Egr1 regulates the HIF-1alpha gene during hypoxia. AB - Using oligonucleotide expression microarrays we have examined the modulation of gene expression in the DU145 prostate cancer cell line. Our findings confirm that the Egr1 transcription factor is rapidly and transiently upregulated by hypoxia. Furthermore, we have demonstrated that HIF-1alpha mRNA is also transiently upregulated, as is its target gene VEGF. To elucidate the mechanism of the transcriptional upregulation of the HIF-1alpha gene, we have shown that Egr1 is able to directly bind to the HIF-1alpha promoter using chromatin immunoprecipitation. We also provide evidence that the binding of Egr1 is necessary for the trans-activation of the HIF-1alpha promoter. These studies highlight the importance for the Egr1 transcription factor in the hypoxic response in cultured prostate cancer cell lines, and indicate that the response of Egr1 is upstream of HIF-1 in these cells. These studies are the first demonstration that the HIF-1alpha transcription factor is targeted directly by Egr1 in hypoxia. PMID- 18506760 TI - Potent cell growth inhibitory effects in hepatitis B virus X protein positive hepatocellular carcinoma cells by the selective cyclooxygenase-2 inhibitor celecoxib. AB - Hepatitis B virus (HBV) X protein (HBx) and cyclooxygenase-2 (COX-2) are all playing roles in hepatocellular carcinoma (HCC), but the reversing effects of COX 2 inhibitors on the neoplastic features caused by HBx protein is still unclear. To further evaluate the therapeutic potential of celecoxib on HBx mediated transformation, HCC cells transfected with HBx gene were treated with COX-2 selective inhibitor, celecoxib. The amount the main metabolite of COX-2, prostaglandin E2 (PGE2), was determined by using high sensitivity ELISA. Electron microscope and flow cytometry was used to analyze cell apoptosis and cell cycle distribution. RT-PCR and Western blot were used to identify the molecules involved in celecoxib induced cell apoptosis. The results showed that celecoxib inhibited cell growth more significantly and also induced more cell apoptosis in HBx over-expression cells than in control cells. Celecoxib could selectively inhibited COX-2 expression and PGE2 production. Celecoxib also inhibited p(473Ser)Akt, raf and p53 expression, and induced apoptosis by release of cytochrome c and activation of caspase 9, 3, and 6, which were more remarkably in HBx positive cells than in control cells. These results suggest that celecoxib had potent cell growth inhibitory effects on HBx positive HCC cells mainly through inducing more cell apoptosis, and these findings provide a new insight into the anticancer effects of celecoxib against HBx related HCC. PMID- 18506762 TI - Anatomy and histochemistry of hindlimb flight posture in birds. I. The extended hindlimb posture of shorebirds. AB - Birds utilize one of two hindlimb postures during flight: an extended posture (with the hip and knee joints flexed, while the ankle joint is extended caudally) or a flexed posture (with the hip, knee, and ankle joints flexed beneath the body). American Avocets (Recurvirostra americana) and Black-necked Stilts (Himantopus mexicanus) extend their legs caudally during flight and support them for extended periods. Slow tonic and slow twitch muscle fibers are typically found in muscles functioning in postural support due to the fatigue resistance of these fibers. We hypothesized that a set of small muscles composed of high percentages of slow fibers and thus dedicated to postural support would function in securing the legs in the extended posture during flight. This study examined the anatomy and histochemical profile of eleven hindlimb muscles to gain insight into their functional roles during flight. Contrary to our hypothesis, all muscles possessed both fast twitch and slow twitch or slow tonic fibers. We believe this finding is due to the versatility of dynamic and postural functions the leg muscles must facilitate, including standing, walking, running, swimming, and hindlimb support during flight. Whether birds use an extended or flexed hindlimb flight posture may be related to the aerodynamic effect of leg position or may reflect evolutionary history. PMID- 18506763 TI - Assigning a cause for a particular outcome in oncology: a serious note of caution. PMID- 18506764 TI - Prognostic factors for patients with Ewing sarcoma (EWS) at first recurrence following multi-modality therapy: A report from the Children's Oncology Group. AB - BACKGROUND: The prognosis for patients with recurrent Ewing sarcoma (EWS) is very poor with 5-year survival of 13%. METHODS: To evaluate prognostic factors for these patients we studied patients initially treated on the multi-institutional study INT0091. RESULTS: Two hundred sixty-two patients experienced disease recurrence. The median time to first recurrence was 1.3 years (0-7.4 years), 1.4 years (0-7.4 years) for patients with initially localized disease and 1 year (0-6 years) for patients with initially metastatic disease. Time to first recurrence from date of initial diagnosis was a predictor of post-recurrence survival (P < 0.0001). Twenty-one percent of patients, with recurrent or progressive disease >or=2 years from initial diagnosis, had an estimated 5-year survival of 30% (vs. 7% estimated 5-year survival with an earlier recurrence). No statistical difference was detected between patients whose disease recurred <1 year and between 1 and 2 years from initial diagnosis. A stepwise relative risk model and backwards stepwise regression was used to explore factors significantly associated with risk for post-recurrence death. Significant risk factors for death after recurrence included recurrence at combined local and distant sites, elevated LDH at initial diagnosis and initial recurrence less than 2 years after diagnosis. Isolated pulmonary recurrence was not predictive of survival after recurrence. CONCLUSION: Patients with a longer disease control interval represent the subset of patients most likely to survive following recurrence of EWS. All patients with recurrence would benefit from collaborative trials to investigate new therapeutic options. PMID- 18506765 TI - Long-term outcome of childhood acute lymphoblastic leukemia treated in China. AB - BACKGROUND: To retrospectively determine the treatment outcome and causes of treatment failure of ALL children treated in a single institution at East China. PROCEDURE: Between January 1998 and October 2004, 346 newly diagnosed ALL patients 0.05 by a Wilcoxon matched pairs test). A methacholine challenge test was also administered to an animal and reduction and recovery of r was detected by the FAVOR method. The reduced 3He consumption and the improvement in speed provided by FAVOR suggest that it may allow measurement of ventilation in human subjects not previously possible. PMID- 18506797 TI - Water spin dynamics during apoptotic cell death in glioma gene therapy probed by T1rho and T2rho. AB - Longitudinal and transverse relaxations in the rotating frame, with characteristic time constants T1rho and T2rho, respectively, have potential to provide unique MRI contrast in vivo. On-resonance spin-lock T1rho with different spin-lock field strengths and adiabatic T2rho with different radiofrequency modulation functions were measured in BT4C gliomas treated with Herpes Simplex Virus thymidine kinase (HVS-tk) gene therapy causing apoptotic cell death. These NMR tools were able to discriminate different treatment responses in tumor tissue from day 4 onward. An equilibrium two-site exchange model was used to calculate intrinsic parameters describing changes in water dynamics. Observed changes included increased correlation time of water associated with macromolecules and a decreased fractional population of this pool. These results are consistent with destructive intracellular processes associated with cell death and the increase of extracellular space during the treatment. Furthermore, association between longer exchange correlation time and decreased pH during apoptosis is discussed. In this study, we demonstrated that T1rho and T2rho MR imaging are useful tools to quantify early changes in water dynamics reflecting treatment response during gene therapy. PMID- 18506798 TI - Mn enhancement and respiratory gating for in utero MRI of the embryonic mouse central nervous system. AB - The mouse is the preferred model organism for genetic studies of mammalian brain development. MRI has potential for in utero studies of mouse brain development, but has been limited previously by challenges of maximizing image resolution and contrast while minimizing artifacts due to physiological motion. Manganese (Mn) enhanced MRI (MEMRI) studies have demonstrated central nervous system (CNS) contrast enhancement in mice from the earliest postnatal stages. The purpose of this study was to expand MEMRI to in utero studies of the embryonic CNS in combination with respiratory gating to decrease motion artifacts. We investigated MEMRI-facilitated CNS segmentation and three-dimensional (3D) analysis in wild type mouse embryos from midgestation, and explored effects of Mn on embryonic survival and image contrast. Motivated by observations that MEMRI provided an effective method for visualization and volumetric analysis of embryonic CNS structures, especially in ventral regions, we used MEMRI to examine Nkx2.1 mutant mice that were previously reported to have ventral forebrain defects. Quantitative MEMRI analysis of Nkx2.1 knockout mice demonstrated volumetric changes in septum (SE) and basal ganglia (BG), as well as alterations in hypothalamic structures. This method may provide an effective means for in utero analysis of CNS phenotypes in a variety of mouse mutants. PMID- 18506799 TI - AxCaliber: a method for measuring axon diameter distribution from diffusion MRI. AB - The diameter of a myelinated nerve axon is directly proportional to its conduction velocity, so the axon diameter distribution helps determine the channel capacity of nervous transmission along fascicles in the central (CNS) and peripheral nervous systems (PNS). Previously, this histological information could only be obtained using invasive tissue biopsies. Here we propose a new NMR-based approach that employs a model of water diffusion within "restricted" cylindrical axons to estimate their diameter distribution within a nerve bundle. This approach can be combined with MRI to furnish an estimate of the axon diameter distribution within each voxel. This method is validated by comparing the diameter distributions measured using the NMR and histological techniques on sciatic and optic nerve tissue specimens. The axon diameter distribution measured in each voxel of porcine spinal cord using MRI and using histological methods were similar. Applications are expected in longitudinal studies designed to follow nerve growth in normal and abnormal development, as well as in diagnosing disorders and diseases affecting specific populations of axons in the CNS and PNS. PMID- 18506800 TI - Fast slice-selective radio-frequency excitation pulses for mitigating B+1 inhomogeneity in the human brain at 7 Tesla. AB - A novel radio-frequency (RF) pulse design algorithm is presented that generates fast slice-selective excitation pulses that mitigate B+1 inhomogeneity present in the human brain at high field. The method is provided an estimate of the B+1 field in an axial slice of the brain and then optimizes the placement of sinc like "spokes" in kz via an L1-norm penalty on candidate (kx, ky) locations; an RF pulse and gradients are then designed based on these weighted points. Mitigation pulses are designed and demonstrated at 7T in a head-shaped water phantom and the brain; in each case, the pulses mitigate a significantly nonuniform transmit profile and produce nearly uniform flip angles across the field of excitation (FOX). The main contribution of this work, the sparsity-enforced spoke placement and pulse design algorithm, is derived for conventional single-channel excitation systems and applied in the brain at 7T, but readily extends to lower field systems, nonbrain applications, and multichannel parallel excitation arrays. PMID- 18506801 TI - Monitoring prostate thermal therapy with diffusion-weighted MRI. AB - For MR-guided minimally invasive therapies, it is important to have a repeatable and reliable tissue viability evaluation method. The use of diffusion-weighted MRI (DWI) to evaluate tissue damage was assessed in 19 canine prostates with cryoablation or high-intensity ultrasound (HIU) ablation. The apparent diffusion coefficient (ADC) trace value was measured in the treated tissue immediately upon the procedure and on the posttreatment follow-up. For the acute lesions, the ADC value decreased to (1.05+/-0.25)x10(-3) mm2/s, as compared to (1.64+/-0.24)x10( 3) mm2/s before the treatment. There was no statistical difference between previously frozen or previously ultrasound-heated lesions in terms of the 36% ADC reduction (P=0.66). The ADC decrease occurred early during the course of the treatment, which appears to complicate DWI-based thermometry. Over time, the ADC value increased as the tissue recovered and regenerated. This study shows that DWI could be a promising method to monitor prostate thermal therapies and to provide insight on tissue damage and tissue remodeling after injury. PMID- 18506802 TI - Single- or dual-bolus approach for the assessment of myocardial perfusion reserve in quantitative MR perfusion imaging. AB - A dual-bolus protocol can overcome limitations due to T1-induced MR signal attenuation and hence enables more accurate quantification of myocardial blood flow (MBF) by contrast enhanced MR perfusion imaging. The study explores potential benefits of the dual-bolus technique for the assessment of myocardial perfusion reserve (MPR) over a standard single-bolus protocol. Nineteen patients without obstructive coronary artery disease as assessed by cardiac catheterization underwent a stress-rest MR perfusion study using a dual-bolus protocol. Gd-DTPA dosages of 0.005 and 0.05 mmol/kg of bodyweight were delivered as pre- and main-bolus. For comparison arterial input curves where extracted from left ventricular cavity passage including both, pre-bolus and main-bolus data. Global and segmental MPR were determined from semiquantitative and from full quantitative measures of MBF. As a result good agreement between dual- and single bolus technique was found with relative differences of maximally 10% in global MPR estimates. For the dual bolus approach a significant relative decrease of 30% (P<0.001) was found for the coefficient of segmental MPR variation, which may allow a more reliable detection of hypoperfused segments in clinical studies. PMID- 18506803 TI - Enhancement of drug solubility in supramolecular and colloidal systems. AB - Statins, as efficient HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme-A) reductase inhibitors are widely used in the management of cardiovascular diseases. Interactions in aqueous solutions between highly lipophilic statins and cyclodextrins (CDs) in the absence and the presence of a dissolved polymer or its monomeric compound, respectively, were studied. The solubility of lovastatin and simvastatin at various temperatures and pHs were investigated by phase-solubility measurements. Surface activity of solutes in binary (CD-statin) and ternary (CD statin-polymer) systems was studied by determining the surface tension of the solutions. For the characterization of the CD-statin inclusion complexes, stability constants for associates of different molar ratios have been calculated. It was shown that complexation may lead to improvement of the aqueous solubilities of both statins by 1-2 orders of magnitude. Especially, randomly methylated beta-cyclodextrin (RAMEB) showed outstanding solubilizing effects. In binary systems dominantly CD-statin associates of 1:1 molar ratios form, which exhibit considerable surface activity. RAMEB forms more stable complexes with these drugs than the native beta-CD, and also the surface activity of the former solutes is higher. In polymer-containing ternary systems the solubility of both statins could be further improved. The enhanced drug solubilities can be ascribed to the formation of CD-statin-polymer associates with supramolecular structure. A portion of the surface active CD-statin complexes are very likely anchored at the macromolecular chains. In these solutions, the total amounts of solutes are composed of the sum of the "free" binary and the supramolecular ternary associates. PMID- 18506805 TI - Improving the pharmacokinetic parameter measurement in dynamic contrast-enhanced MRI by use of the arterial input function: theory and clinical application. AB - One of the most powerful features of the dynamic contrast-enhanced (DCE) MRI technique is its capability to quantitatively measure the physiological or pathophysiological environments assessed by the passage of contrast agent by means of model-based pharmacokinetic analysis. The widely used two-compartment pharmacokinetic model developed by Brix and colleges fits tumor data well in most cases, but fails to explain the biexponential arterial input function. In this work, this problem has been attacked from a theoretical point of view, showing that this problem can be solved by adopting a more realistic model assumption when simplifying the general solutions of the two-compartment pharmacokinetic equations. Pharmacokinetic parameters derived from our model were demonstrated to have comparative tissue specificity to Ktrans from Larsson's model, better than those from Brix's model and the empirical area-under-the-curve (AUC). Tissue-type classifier constructed with the arterial input function-decomposed kep-kpe pair from our model was also demonstrated to have superior performance than any other classifier based on DCE-MRI pharmacokinetic parameters or empirical AUC. The feature that this classifier has a near-zero false-negative rate makes it a highly desirable tool for clinical diagnostic and response assessment applications. PMID- 18506804 TI - Ultrasonic-activated micellar drug delivery for cancer treatment. AB - The use of nanoparticles and ultrasound in medicine continues to evolve. Great strides have been made in the areas of producing micelles, nanoemulsions, and solid nanoparticles that can be used in drug delivery. An effective nanocarrier allows for the delivery of a high concentration of potent medications to targeted tissue while minimizing the side effect of the agent to the rest of the body. Polymeric micelles have been shown to encapsulate therapeutic agents and maintain their structural integrity at lower concentrations. Ultrasound is currently being used in drug delivery as well as diagnostics, and has many advantages that elevate its importance in drug delivery. The technique is noninvasive, thus no surgery is needed; the ultrasonic waves can be easily controlled by advanced electronic technology so that they can be focused on the desired target volume. Additionally, the physics of ultrasound are widely used and well understood; thus ultrasonic application can be tailored towards a particular drug delivery system. In this article, we review the recent progress made in research that utilizes both polymeric micelles and ultrasonic power in drug delivery. PMID- 18506806 TI - Minimizing acquisition time of arterial spin labeling at 3T. AB - An improved arterial spin labeling (ASL) perfusion technique that combines pseudo continuous labeling and a T2*-insensitive sequence (GRASE) with background suppression was used to acquire perfusion maps in normal volunteers and stroke patients. It is shown that perfusion measurements obtained in less than 1 min of scan time are reproducible, with a coefficient of variation of 7%. The perfusion maps generated from these data can be used to characterize the stroke lesion. PMID- 18506807 TI - New techniques for cartilage magnetic resonance imaging relaxation time analysis: texture analysis of flattened cartilage and localized intra- and inter-subject comparisons. AB - MR relaxation time measurements of knee cartilage have shown potential to characterize knee osteoarthritis (OA). In this work, techniques that allow localized intra- and inter-subject comparisons of cartilage relaxation times, as well as cartilage flattening for texture analysis parallel and perpendicular to the natural cartilage layers, are presented. The localized comparisons are based on the registration of bone structures and the assignment of relaxation time feature vectors to each point in the bone-cartilage interface. Cartilage flattening was accomplished with Bezier splines and warping, and texture analysis was performed with second-order texture measures using gray-level co-occurrence matrices (GLCM). In a cohort of five normal subjects the performance and reproducibility of the techniques were evaluated using T1rho maps of femoral knee cartilage. The feasibility of creating a mean cartilage relaxation time map is also presented. Successful localized intra- and inter-subject T1rho comparisons were obtained with reproducibility similar to that reported in the literature for regional T2. Improvement of the reproducibility of GLCM features was obtained by flattening the T1rho maps. The results indicate that the presented techniques have potential in longitudinal and population studies of knee OA at different stages of the disease. PMID- 18506809 TI - Gel formation and low-temperature intramolecular conformation transition of a triple-helical polysaccharide lentinan in water. AB - The gelation behavior of the triple-helical polysaccharide lentinan fractions having different molecular weights in water at 25 degrees C were studied by using a rheometer. The analysis of concentration and molecular weight dependence of shear stress and shear viscosity showed that aqueous lentinan is a typical shear thinning fluid, possessing potential as a viscosity control agent, and that a weak gel with entangled network structure formed. The dynamic oscillatory behavior of lentinan in the temperature range of 1-15 degrees C was also investigated by rheologic method. The storage modulus G' and complex viscosity eta* increased first with decreasing temperature, and underwent a maximum centered at 7-9 degrees C, and then decreased with further decreasing temperature. This abnormal phenomenon was ascribed to formation of rigid structure in the gel state, which was confirmed by the experimental results from micro-DSC. The micro-DSC curves showed that an endothermic peak appeared at 7-9 degrees C for lentinan in water upon heating, which was attributable to the intramolecular order-disorder structure transition similar to triple-helical polysaccharide schizophyllan. Namely, at lower temperature, the side glucose residues of lentinan (triplix II) formed a well-organized triple-helical structure (triplix I) through hydrogen-bonding with the surrounding water molecules. Moreover, this conformation transition was proved to be thermally reversible. PMID- 18506810 TI - Solvent interactions with the Trp-cage peptide in 35% ethanol-water. AB - Intermolecular NOE experiments have been used to explore interactions of water and ethanol molecules in 35% ethanol/65% water (v/v) with the peptide Trp-cage at temperatures from 5 to 25 degrees C. Magnetic dipole-dipole cross-relaxation terms sigma(HH) (NOE) and sigma(HH) (ROE) for interaction of solvent components with spins of the peptide suggest that ethanol molecules associate with backbone atoms for times of the order of nanoseconds at 5 degrees C. Formation of peptide ethanol complexes can also account for the larger-than-expected values of cross relaxation terms at higher temperatures. Hydrocarbon side chains of the peptide do not appear to experience such interactions with ethanol. Cross relaxation resulting from water-peptide interactions are consistent with long-lived water interactions with the backbone atoms. Water cross relaxation with nonpolar side chains of the peptide (Leu2, Ile4, Leu7, and proline residues) are only those expected for bulk solvent. However, long-lived association of both water and ethanol with the polar side chains of Tyr3 and Trp6 is indicated by the data. PMID- 18506808 TI - ABSINTH: a new continuum solvation model for simulations of polypeptides in aqueous solutions. AB - A new implicit solvation model for use in Monte Carlo simulations of polypeptides is introduced. The model is termed ABSINTH for self-Assembly of Biomolecules Studied by an Implicit, Novel, and Tunable Hamiltonian. It is designed primarily for simulating conformational equilibria and oligomerization reactions of intrinsically disordered proteins in aqueous solutions. The paradigm for ABSINTH is conceptually similar to the EEF1 model of Lazaridis and Karplus (Proteins 1999, 35, 133). In ABSINTH, the transfer of a polypeptide solute from the gas phase into a continuum solvent is the sum of a direct mean field interaction (DMFI), and a term to model the screening of polar interactions. Polypeptide solutes are decomposed into a set of distinct solvation groups. The DMFI is a sum of contributions from each of the solvation groups, which are analogs of model compounds. Continuum-mediated screening of electrostatic interactions is achieved using a framework similar to the one used for the DMFI. Promising results are shown for a set of test cases. These include the calculation of NMR coupling constants for short peptides, the assessment of the thermal stability of two small proteins, reversible folding of both an alpha-helix and a beta-hairpin forming peptide, and the polymeric properties of intrinsically disordered polyglutamine peptides of varying lengths. The tests reveal that the computational expense for simulations with the ABSINTH implicit solvation model increase by a factor that is in the range of 2.5-5.0 with respect to gas-phase calculations. PMID- 18506811 TI - Multiaxial analysis of dental composite materials. AB - Dental composites are subjected to extreme chemical and mechanical conditions in the oral environment, contributing to the degradation and ultimate failure of the material in vivo. The objective of this study is to validate an alternative method of mechanically loading dental composite materials. Confined compression testing more closely represents the complex loading that dental restorations experience in the oral cavity. Dental composites, a nanofilled and a hybrid microfilled, were prepared as cylindrical specimens, light-cured in ring molds of 6061 aluminum, with the ends polished to ensure parallel surfaces. The samples were subjected to confined compression loading to 3, 6, 9, 12, and 15% axial strain. Upon loading, the ring constrains radial expansion of the specimen, generating confinement stresses. A strain gage placed on the outer wall of the aluminum confining ring records hoop strain. Assuming plane stress conditions, the confining stress (sigma(c)) can be calculated at the sample/ring interface. Following mechanical loading, tomographic data was generated using a high resolution microtomography system developed at beamline 2-BM of the Advanced Photon Source at Argonne National Laboratory. Extraction of the crack and void surfaces present in the material bulk is numerically represented as crack edge/volume (CE/V), and calculated as a fraction of total specimen volume. Initial results indicate that as the strain level increases the CE/V increases. Analysis of the composite specimens under different mechanical loads suggests that microtomography is a useful tool for three-dimensional evaluation of dental composite fracture surfaces. PMID- 18506812 TI - Intracerebral microinjection of stannous 2-ethylhexanoate affects dopamine turnover in cerebral cortex and locomotor activity in rats. AB - Stannous 2-ethylhexanoate [Sn(Oct)(2)] is used as a catalyst for production of poly-L-lactic acid and copolymers that are implanted in cranial surgery, but reports on its effects on the central nervous system are few. We examined the effects of Sn(Oct)(2) on cell viability in vitro and on neurotransmission and behavior in the rat. Treatment of normal human astrocytes with 10 mg/mL Sn(Oct)(2) reduced mitochondrial activity to 16% of the control. Injection of Sn(Oct)(2) at 6.28 mg/kg BW (2 mg/kg BW Sn) into right lateral ventricle of the rat brain tended to increase the ambulation distance after 30 days when compared with the control group. The turnover of dopamine neurotransmission was increased in the cerebral cortex. These results suggest that Sn(Oct)(2) is cytotoxic to astrocytes in vitro. Injection of Sn(Oct)(2) into the brain had no or very weak immediate neurotoxicity, but long-term exposure to Sn(Oct)(2) increased dopamine neurotransmission turnover. PMID- 18506813 TI - Fabrication, characterization, and biocompatibility of single-walled carbon nanotube-reinforced alginate composite scaffolds manufactured using freeform fabrication technique. AB - Composite polymeric scaffolds from alginate and single-walled carbon nanotube (SWCNT) were produced using a freeform fabrication technique. The scaffolds were characterized for their structural, mechanical, and biological properties by scanning electron microscopy, Raman spectroscopy, tensile testing, and cell scaffold interaction study. Three-dimensional hybrid alginate/SWCNT tissue scaffolds were fabricated in a multinozzle biopolymer deposition system, which makes possible to disperse and align SWCNTs in the alginate matrix. The structure of the resultant scaffolds was significantly altered due to SWCNT reinforcement, which was confirmed by Raman spectroscopy. Microtensile testing presented a reinforcement effect of SWCNT to the mechanical strength of the alginate struts. Ogden constitutive modeling was utilized to predict the stress-strain relationship of the alginate scaffold, which compared well with the experimental data. Cellular study by rat heart endothelial cell showed that the SWCNT incorporated in the alginate structure improved cell adhesion and proliferation. Our study suggests that hybrid alginate/SWCNT scaffolds are a promising biomaterial for tissue engineering applications. PMID- 18506814 TI - Photocatalytic and antibacterial properties of medical-grade PVC material coated with TiO2 film. AB - The TiO(2) film was coated on poly vinyl chloride (PVC) surface by dip-coating process from TiO(2)-PVC-THF suspension. The morphology and crystal structure of the as-synthesized samples were characterized by SEM and XRD. The photocatalytic properties were measured by the photodegradation reaction of RhB and the anti adhesion and anti-bacteria for Escherichia coli. The results show that the resultant TiO(2) film is well-conglutinated on PVC surface and has the same crystal structure as the original TiO(2) powder. The TiO(2)/PVC shows excellent photocatalytic activity for the degradation of aqueous RhB and the activity increases with increasing reaction time and tends toward stable after accumulative illumination for 11.5 h. The TiO(2) film shows good bacterial anti adhesion activity following photo-activation and sterilization property under UV irradiation. The E. coli can be killed completely after UV irradiation for 1.5 h. PMID- 18506815 TI - Hydroxylation of dental zirconia surfaces: characterization and bonding potential. AB - Bioinert zirconia surfaces exhibit a low chemical bonding potential to resin based luting agents. The aim was to hydroxylate dental zirconia surfaces and to examine tensile bond strength using commercial luting agents. The measured bond strength was compared with established mechanical conditioning techniques. Five acidic and one alkaline hydroxylation pretreatments were applied and compared with air abrasion and tribochemical silica coating. For the chemical characterization of hydroxyl groups and hydroxyl value, zirconia powders were used, chemically modified, and analyzed using Fourier-transformed infrared spectroscopy and a titrimetric method according to the ISO 4629 standard. All acidic pretreatment procedures exhibited increased hydroxyl values. The highest values were recorded after treatment with phosphoric acid or Piranha solution. Tensile bond strength was examined in a universal testing machine using the commercial dual-cure luting agents Multilink (Ivoclar, Liechtenstein) and Panavia F2.0 (Kuraray, Japan). Surface hydroxylation with Piranha solution in combination with the luting agent Multilink led to a bond strength of 12.47 +/- 3.25 MPa. Tribochemical silica-coated/silanized zirconia surfaces with Multilink produced the highest bond strength of 19.33 +/- 3.65 MPa. Using the luting agent Panavia F2.0, statistically homogenous values for the untreated (11.60 +/- 1.68 MPa) and for the hydroxylated surface (12.46 +/- 3,81 MPa) were measured. Bioinert zirconia surfaces were successfully hydroxylated in terms of tensile bond strength. Resin bonding with Multilink can be strongly increased by acidic treatment with Piranha solution. Bonding with Panavia F2.0 is not affected by hydroxylation, which is likely due to the incorporation of specific functional monomers. PMID- 18506816 TI - Remaining unreacted methacrylate groups in resin-based composite with respect to sample preparation and storing conditions using micro-Raman spectroscopy. AB - The aim of this study was to measure degree of conversion (DC) of resin-based composites (RBCs) using micro-Raman spectroscopy followed by different sample preparation procedures and storing conditions. Ninety samples of Tetric EvoCeram (Ivoclar Vivadent, Schaan, Liechtenstein) were prepared in standardized molds and cured with a high powered LED light-curing unit, bluephase (Ivoclar Vivadent, Schaan, Liechtenstein) for 20 s. Samples were allocated to eight groups. DC of groups 1 and 2 was recorded without or after polishing. DC in groups 3 and 4 was recorded from vertically sectioned samples versus "split" samples. DC in groups 5 8 was recorded after storing samples at room temperature and humidity, in 90 +/- 2% humidity at 37 +/- 1 degrees C, distilled water at 37 +/- 1 degrees C or buffered incubation medium (BIM) at 37 +/- 1 degrees C for 24 h. Mean values of DC in polished and unpolished samples were 63.6% (+/-3.2%) and 54.7% (+/-5.2%), respectively (p < 0.0001). There was no significant difference in DC after sample sectioning (p > 0.05). Significantly higher DC values were obtained after storing samples in BIM (76.8% +/- 2.1%) than in distilled water (59.7% +/- 5.7%), extreme humidity (60.3% +/- 3.9%) or in room conditions (63.6% +/- 3.2%) (p < 0.001). DC of an RBC measured by micro-Raman spectroscopy may be affected by differences in sample preparation and storing conditions, making it difficult to extrapolate data from in vitro studies into clinically relevant information. PMID- 18506817 TI - Relationship between P-glycoprotein activity measured in peripheral blood mononuclear cells and indinavir bioavailability in healthy volunteers. AB - Indinavir, a HIV-1 protease inhibitor, showed large inter-individual pharmacokinetic variability. It has been proposed as a substrate of P glycoprotein (P-gp), an efflux transporter, that may contribute to limit indinavir bioavailability. A liquid formulation of indinavir was developed from indinavir capsules in order to study indinavir pharmacokinetics in healthy volunteers. Compartmental and noncompartmental analysis of indinavir plasma concentrations showed high inter-individual variability in terms of area under the curve (AUC) and maximal plasma concentration (C(max)). A significant negative association between AUC normalized to body weight (AUC x weight) and lymphocyte P gp activity, using Rh123 efflux assay, was observed (p = 0.008; r = -0.75). AUC normalized to elimination rate constant (AUC x beta) also showed a significant negative relationship with lymphocyte P-gp activity (p = 0.03, r = -0.64). Apparent clearance (CL/[F x weight]) and volume of distribution (VD/[F x weight]) showed a positive correlation with P-gp activity. Conversely, elimination rate constant did not correlate with P-gp activity. Although there is not enough evidence of a correlation between lymphocitary and intestinal function of P-gp, our results suggest a relationship between a P-gp phenotype marker, Rh123 efflux assay in lymphocytes, and indinavir bioavailability. PMID- 18506818 TI - Rat nasal lavage biomarkers to assess preclinical irritation potential of nasal drug formulations and excipients. AB - The goal of this study was to evaluate biomarkers of nasal mucosal damage for rapid assessment of irritancy potential of formulations in the rat nasal lavage model, a tool to facilitate nasal formulation development prior to histopathology studies. The nasal cavity of anesthetized rats was lavaged with normal saline 20 min pos-tdose. The collected fluid was analyzed for secreted total protein and biomarkers. Solutions tested include: normal saline, buffers, benzalkonium chloride (BAC), lysophosphatidylcholine (LPC), and four marketed nasal products. Total protein, lactate dehydrogenase and interleukin-1alpha biomarkers were secreted to varying degrees. BAC (0.2%) and LPC (0.5%) exhibiting the strongest response with a signal window ranging from 3.4- to 87-fold greater levels than normal saline. Buffer treatments, excipients, and most marketed nasal products yielded levels similar to normal saline. There was a weak correlation between formulation osmolarity and surface tension with any of the biomarkers. Each nasal formulation elicited a unique protein and biomarker profile with total protein secretion correlated with IL-1alpha secretion suggesting the potential for an inflammatory response. Taken together, rapid and potentially mechanistic information on the preclinical acute irritancy potential of formulations was assessed in the rat nasal lavage model by benchmarking treatments relative to controls and marketed nasal products. PMID- 18506819 TI - Dry powder formulations for inhalation of fluticasone propionate and salmeterol xinafoate microcrystals. AB - Direct crystallization of active pharmaceutical ingredient (API) particles in the inhalable size range of 1-6 microm may overcome surface energization resulting from micronization. The aerosolization of fluticasone propionate (FP) and salmeterol xinafoate (SX) microcrystals produced by aqueous crystallization from poly(ethylene glycol) solutions was investigated using a twin stage impinger following blending with lactose. Fine particle fractions from SX formulations ranged from 15.98 +/- 2.20% from SX crystallized from PEG 6000 to 26.26 +/- 1.51% for SX crystallized from PEG 400. The FPF of microcrystal formulations increased as the particle size of microcrystals was increased. The aerosolization of SX from DPI blends was equivalent for the microcrystals and the micronized material. FP microcrystals, which had a needlelike morphology, produced similar FPFs (PEG 400: 17.15 +/- 0.68% and PEG 6000: 15.46 +/- 0.97%) to micronized FP (mFP; 14.21 +/- 0.54). The highest FPF (25.66 +/- 1.51%) resulted from the formulation of FP microcrystals with the largest median diameter (FP PEG 400B: 6.14 +/- 0.17 microm). Microcrystallization of SX and FP from PEG solvents offers the potential for improving control of particulate solid state properties and was shown to represent a viable alternative to micronization for the production of particles for inclusion in dry powder inhalation formulations. PMID- 18506820 TI - A procedure to optimize scale-up for the primary drying phase of lyophilization. AB - This article describes a procedure to facilitate scale-up for the primary drying phase of lyophilization using a combination of empirical testing and numerical modeling. Freeze dry microscopy is used to determine the temperature at which lyophile collapse occurs. A laboratory scale freeze-dryer equipped with manometric temperature measurement is utilized to characterize the formulation dependent mass transfer resistance of the lyophile and develop an optimized laboratory scale primary drying phase of the freeze-drying cycle. Characterization of heat transfer at both lab and pilot scales has been ascertained from data collected during a lyophilization cycle involving surrogate material. Using the empirically derived mass transfer resistance and heat transfer data, a semi-empirical computational heat and mass transfer model originally developed by Mascarenhas et al. (Mascarenhas et al., 1997, Comput Methods Appl Mech Eng 148: 105-124) is demonstrated to provide predictive primary drying data at both the laboratory and pilot scale. Excellent agreement in both the sublimation interface temperature profiles and the time for completion of primary drying is obtained between the experimental cycles and the numerical model at both the laboratory and pilot scales. Further, the computational model predicts the optimum operational settings of the pilot scale lyophilizer, thus the procedure discussed here offers the potential to both reduce the time necessary to develop commercial freeze-drying cycles by eliminating experimentation and to minimize consumption of valuable pharmacologically active materials during process development. PMID- 18506821 TI - Neural agrin increases postsynaptic ACh receptor packing by elevating rapsyn protein at the mouse neuromuscular synapse. AB - Fluorescence resonance energy transfer (FRET) experiments at neuromuscular junctions in the mouse tibialis anterior muscle show that postsynaptic acetylcholine receptors (AChRs) become more tightly packed during the first month of postnatal development. Here, we report that the packing of AChRs into postsynaptic aggregates was reduced in 4-week postnatal mice that had reduced amounts of the AChR-associated protein, rapsyn, in the postsynaptic membrane (rapsyn(+/-) mice). We hypothesize that nerve-derived agrin increases postsynaptic expression and targeting of rapsyn, which then drives the developmental increase in AChR packing. Neural agrin treatment elevated the expression of rapsyn in C2 myotubes by a mechanism that involved slowing of rapsyn protein degradation. Similarly, exposure of synapses in postnatal muscle to exogenous agrin increased rapsyn protein levels and elevated the intensity of anti-rapsyn immunofluorescence, relative to AChR, in the postsynaptic membrane. This increase in the rapsyn-to-AChR immunofluorescence ratio was associated with tighter postsynaptic AChR packing and slowed AChR turnover. Acute blockade of synaptic AChRs with alpha-bungarotoxin lowered the rapsyn-to-AChR immunofluorescence ratio, suggesting that AChR signaling also helps regulate the assembly of extra rapsyn in the postsynaptic membrane. The results suggest that at the postnatal neuromuscular synapse agrin signaling elevates the expression and targeting of rapsyn to the postsynaptic membrane, thereby packing more AChRs into stable, functionally-important AChR aggregates. PMID- 18506822 TI - Differential expression of connexins during histogenesis of the chick retina. AB - Gap junction (GJ) channels couple adjacent cells, allowing transfer of second messengers, ions, and molecules up to 1 kDa. These channels are composed by a multigene family of integral membrane proteins called connexins (Cx). In the retina, besides being essential circuit element in the visual processing, GJ channels also play important roles during its development. Herein, we analyzed Cx43, Cx45, Cx50, and Cx56 expression during chick retinal histogenesis. Cx exhibited distinct expression profiles during retinal development, except for Cx56, whose expression was not detected. Cx43 immunolabeling was observed at early development, in the transition of ventricular zone and pigmented epithelium. Later, Cx43 was seen in the outer plexiform and ganglion cell layers, and afterwards also in the inner plexiform layer. We observed remarkable changes in the phosphorylation status of this protein, which indicated modifications in functional properties of this Cx during retinal histogenesis. By contrast, Cx45 showed stable gene expression levels throughout development and ubiquitous immunoreactivity in progenitor cells. From later embryonic development, Cx45 was mainly observed in the inner retina, and it was expressed by glial cells and neurons. In turn, Cx50 was virtually absent in the chick retina at initial embryonic phases. Combination of PCR, immunohistochemistry and Western blot indicated that this Cx was present in differentiated cells, arising in parallel with the formation of the visual circuitry. Characterization of Cx expression in the developing chick retina indicated particular roles for these proteins and revealed similarities and differences when compared to other species. PMID- 18506823 TI - Repeated neonatal separation stress alters the composition of neurochemically characterized interneuron subpopulations in the rodent dentate gyrus and basolateral amygdala. AB - Emotional experience during early life has been shown to interfere with the development of excitatory synaptic networks in the prefrontal cortex, hippocampus, and the amygdala of rodents and primates. The aim of the present study was to investigate a developmental "homoeostatic synaptic plasticity" hypothesis and to test whether stress-induced changes of excitatory synaptic composition are counterbalanced by parallel changes of inhibitory synaptic networks. The impact of repeated early separation stress on the development of two GABAergic neuronal subpopulations was quantitatively analyzed in the brain of the semiprecocial rodent Octodon degus. Assuming that PARV- and CaBP-D28k expression are negatively correlated to the level of inhibitory activity, the previously described reduced density of excitatory spine synapses in the dentate gyrus of stressed animals appears to be "amplified" by elevated GABAergic inhibition, reflected by reduced PARV- (down to 85%) and CaBP-D28k immunoreactivity (down to 74%). In opposite direction, the previously observed elevated excitatory spine density in the CA1 region of stressed animals appears to be amplified by reduced inhibition, reflected by elevated CaPB-D28k immunoreactivity (up to 149%). In the (baso)lateral amygdala, the previously described reduction of excitatory spine synapses appears to be "compensated" by reduced inhibitory activity, reflected by dramatically elevated PARV- (up to 395%) and CaPB-D28k-immunoreactivity (up to 327%). No significant differences were found in the central nucleus of the amygdala, the piriform, and somatosensory cortices and in the hypothalamic paraventricular nucleus. Thus during stress-evoked neuronal and synaptic reorganization, a homeostatic balance between excitation and inhibition is not maintained in all regions of the juvenile brain. PMID- 18506824 TI - Impaction allografting--the effect of impaction force and alternative compaction methods on the mechanical characteristics of the graft. AB - Revision total hip replacement with impaction allografting has an attractive potential for restoring bone stock, however, fractures and implant migration remain problematic. Postoperative graft deformation is believed to contribute to migration. Under compressive loading, the fluid in the graft takes up the load initially through fluid pressure that dissipates over time. Given the short duration of an impaction, we proposed two novel graft compaction techniques that allow more time for fluid flow: holding a compression force constant for 90 s (creep technique), and cycles of compressing the graft to a given force and then holding the displacement constant (cyclic relaxation technique). This study examined the effect of the impaction force on the density and mechanical characteristics of the graft, and explored the potential benefit of the proposed alternative compaction techniques. Increasing the impaction force from "low" to "high" increased graft density by 41%, and this translated to stiffness and shear strength increases of 93 and 164%, respectively. The creep technique improved the stiffness and shear strength by 14 and 16%, respectively, when compared with impaction, while the cyclic relaxation technique did not improve the mechanical properties. Although the creep technique could potentially provide a lower risk of intraoperative fracture over the use of larger impaction forces, any clinical benefit of the creep technique warrants further investigation. Our results point to the importance of maximizing impaction forces in impaction allografting surgery. PMID- 18506825 TI - Bisphosphonate-laden acrylic bone cement: mechanical properties, elution performance, and in vivo activity. AB - Cemented total hip replacements generally fail after 10-20 years, often due to implant loosening from bone resorption. Bisphosphonates such as zoledronic acid (ZA) and pamidronate (PAM) are potent inhibitors of bone resorption. The local delivery of bisphosphonates via acrylic bone cement could decrease osteolysis and prolong implant lifespan. Conflicting studies suggest that bisphosphonate loading may or may not reduce the mechanical properties of acrylic bone cement. We assayed acrylic bone cement laden with ZA or PAM at different concentrations and diluent volumes. Four-point bend testing and compressive testing indicated that high volumes of diluent (with or without bisphosphonate) significantly reduced bending modulus and compressive strength. Radiography and electron microscopy indicated that high diluent volumes generated abnormal acrylic bone cement structure. After 6 weeks of incubation in saline, only 0.9% w/w of the total bisphosphonate incorporated in acrylic bone cement eluted in vitro, indicating a slow elution rate. In vivo testing was performed using a rat model. Cement cylinders were inserted into incisions in rat distal femora and ZA delivered locally (via elution from acrylic bone cement) or systemically (via injection). At 4 weeks postoperatively, dual energy X-ray absorptiometry demonstrated no significant increase in local bone mineral density (BMD) adjacent to ZA-laden implants. In contrast, systemic ZA delivery (0.1 mg/kg) led to a large (48.6%) and significant increase in BMD. Thus, systemic delivery appears more effective than local delivery. PMID- 18506826 TI - The distribution of crystalline material in obstructed stents--in need for intra luminal surface modification? AB - PURPOSE: Usually, hampered urine flow and failing of Seldinger technique leads to the explanation "obstructed ureteral stent" with no further clarification where exactly the obstructions are located. If stent obstruction is caused by intra luminal biofilm and/or crystal deposits, the need of biofilm reducing coatings on the stent's inside has to be discussed. MATERIALS AND METHODS: We investigated 59 stents from patients in whom acute hydronephrosis and/or acute pyelonephritis required stent replacement and/or usage of Seldinger technique failed. The stents were investigated by X-ray and, after longitudinal cutting, by light-microscopy for occurrence of obstructing material. RESULTS: The inside of 25% of the samples was lined with a thick film composed of blood clots and tiny non-aggregated crystals. Only in these samples X-ray investigation showed a positive result for massive inner encrustations, which in fact may be responsible for stent occlusion. 48% of the stents contained few small domains composed of blood clots and crystals. 27% of the stent samples showed no alteration. CONCLUSIONS: 75% of the "obstructed" stents showed no significant inner deposits. Obstruction of urine transport and failure of Seldinger technique occurred due to other reasons. Thus, coating of the stent's inner surface may be overrated. PMID- 18506827 TI - Effects of geometry on fracture initiation and propagation in all-ceramic crowns. AB - The complex and patient-unique geometry of posterior all-ceramic dental crowns represents a particularly interesting set of challenges to understanding stress concentration and fracture evolution in response to loading. A series of numerical and physical experiments, with both single cycle and fatigue loading, show that geometry profoundly influences the stress concentration and fracture initiation and propagation. In stylized crowns with uniform axial wall height, stresses concentrate beneath the indenter. As the height of the axial wall increases, loads to cause failure increase linearly. In crowns with variation in axial wall height around the periphery, stresses concentrate both beneath the indenter and at the margin of the core ceramic. The magnitude of the stress concentration at the margin is directly related to the amount of variation in axial wall height around the periphery of the crown. Anatomically correct veneered zirconia core crowns subjected to single-cycle loads, fracture in areas of greatest stress concentration identified by finite element models. Fractures and stress concentrations that occur in response to single-cycle loading are important indicators of initiation sites for fatigue failure. (c) 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009. PMID- 18506828 TI - Interfacial morphology and bond strength of self-etching adhesives to primary dentin with or without acid etching. AB - The aim of the study was to determine the interfacial morphology and bond strength of three current self-etching adhesives (SEAs) to primary dentin and to evaluate the effect of introducing an additional step of phosphoric acid etching. Three human primary molars were assigned to each adhesive group for testing microtensile bond strength (microTBS) and three for studying interface morphology. Groups were: group 1, Excite, a total-etch adhesive (control); group 2, Adhese (ASE); group 3, Adper-Prompt-L-Pop (APLP), and group 4: Xeno III (XE) SEAs; groups 5-7 received application of 37% phosphoric acid for 15 s before applying ASE, APLP, and XE, respectively. A class I cavity was performed in each molar to study the interface morphology. Two halves of each tooth were used for examination either by optical microscopy, using Masson's trichromic dye technique, or by scanning electron microscopy. For microTBS determination, composite/dentin bars (1 mm(2) section) were obtained from each tooth, and tested in tension until debonding. The microTBS was significantly lower in the APLP than in the rest of the groups. The performance of SEAs on primary dentin depends on the product. Inclusion of dentin pre-etching step did not significantly modify microTBS results. All SAEs achieved greater decalcification depth on etched versus nonetched dentin. PMID- 18506829 TI - Analysis of the fracture morphology of polyamide, polyester, polypropylene, and silk sutures before and after implantation in vivo. AB - This study has analyzed fracture morphology of four nonabsorbable commercially available sutures before and after implantation in rats. Also, tensile and knot strength retention have been evaluated after 3 and 8 weeks in vivo. Scanning electron and optical light microscopes were used for analysis of fracture morphologies and surface changes of the sutures. A clear effect of in vivo on the tensile and knot strength changes, and the fracture mechanism was seen for braided sutures. The suture size was also important for braided sutures, as fibrous tissue formation plays an important role in terms of the size. The fiber's surface properties were also important for the fracture morphology. A smooth and even surface was not suitable for the fibrous tissue formation as seen in monofilament sutures. Therefore, the polymer type was very important for the monofilament sutures, as it was the most important parameter to determine the fracture morphology and was not affected by the implantation and the implantation time. The size of the suture was also important for the polypropylene in terms of axial splitting before and after implantation. This, however, was not the case for knot strength tests. The knot was undone regardless of the size. Surface characteristics were very important for braided sutures, as they have a rough surface that supports tissue formation on the fracture mechanism, tensile and knot strength. Silk suture has single filaments with no regular diameter and smooth surface. Therefore, silk suture has more tissue formation postimplantation compared to polyester suture. PMID- 18506830 TI - Effects of environmental calcium and phosphate on wear and strength of glass ionomers exposed to acidic conditions. AB - This study evaluated the effects of environmental calcium and phosphate on wear resistance, strength, and surface morphology of highly viscous glass-ionomers (HVGICs) (Fuji IX Fast [FN] and KetacMolar [KM]) when exposed to acidic conditions. Fabricated specimens were randomly divided into five groups and kept in acidic solutions (pH 3) with varied levels of calcium and phosphate ranging from 0 to 2.4 mM. After 4 weeks of conditioning, the specimens were subjected to wear testing, shear punch, and surface roughness testing as well as SEM evaluation. Multiple comparisons of wear depth (microm), shear strength (MPa), and surface roughness (Ra) between acidic conditions were performed using ANOVA/post-hoc Scheffe's test (p < 0.05). Results showed that FN and KM exposed to acidic conditions had varied wear resistance, shear strength, surface roughness, and structure depending on environmental phosphate level. Increased level of environmental phosphate led to rougher surface, greater wear resistance, and strength of FN and KM than the controls (acid of pH 3). Under SEM, the surface of both FN and KM specimens were covered by numerous small particles when environmental phosphate was high. Results suggest that environmental phosphate may improve wear resistance and shear strength of HVGICs when challenged by acids. PMID- 18506831 TI - Influence of protein conformation and adjuvant aggregation on the effectiveness of aluminum hydroxide adjuvant in a model alkaline phosphatase vaccine. AB - The mechanism(s) of the enhancement of the immune response by addition of aluminum salt adjuvants to parenterally administered protein-based vaccines is still the subject of debate. It has been hypothesized, however, that destabilization of the antigen structure on the surface of the adjuvant may be important for eliciting immune response. Also, it has been suggested that immune response to adjuvanted vaccines is reduced if the adjuvant particles become aggregated before administration because of processing steps such as freeze drying. In this study, we tested these hypotheses and examined the immune response in a murine model to various liquid, freeze-dried, and spray freeze dried formulations of a model vaccine, bovine intestinal alkaline phosphatase adsorbed on aluminum hydroxide. Enzymatic activity of the alkaline phosphatase was used as a sensitive indicator of intact native antigen structure. By manipulating the secondary drying temperature during lyophilization, vaccines were produced with varying levels of alkaline phosphatase enzymatic activity and varying degrees of adjuvant aggregation, as assessed by particle size distribution. Anti-alkaline phosphatase titers observed in immunized mice were independent of both the antigen's retained enzymatic activity and the vaccine formulation's mean particle diameter. PMID- 18506832 TI - Vibrational circular dichroism and IR spectral analysis as a test of theoretical conformational modeling for a cyclic hexapeptide. AB - A model cyclohexapeptide, cyclo-(Phe-(D)Pro-Gly-Arg-Gly-Asp) was synthesized and its IR and VCD spectra were used as a test of density functional theory (DFT) level predictions of spectral intensities for a peptide with a nonrepeating but partially constricted conformation. Peptide structure and flexibility was estimated by molecular dynamics (MD) simulations and the spectra were simulated using full quantum mechanical (QM) approaches for the complete peptide and for simplified models with truncated side chains. After simulated annealing, the backbone conformation of the ring structure is relatively stable, consisting of a normal beta-turn and a tight loop (no H-bond) which does not vary over short trajectories. Only in quite long MD runs at high temperatures do other conformations appear. MD simulations were carried out for the cyclic peptide in water and in TFE, which match experimental solvents, as well as with and without protonation of the Asp carboxyl group. DFT spectral simulations were made using the annealed structure and were extended to include basis set variation, to determine an optimal computational approach, and solvent simulation with a polarized continuum model (PCM). Stepwise full DFT simulation of spectra was done for various sequences with the same backbone geometry but based on (1) solely Gly residues, (2) Ala substitution except Gly and Pro, and (3) complete sequences with side chains. Additionally, a selection of structures was used to compute IR and VCD spectra with the optimal method to determine structural variation effects. The side chains, especially the Asp-COOH and Arg-NH(2) transitions, had an impact on the computed amide frequencies, IR intensities and VCD pattern. Since experimentally these groups would have little chirality, due to conformational variation, they do not impact the observed VCD spectra. Correcting for frequency shifts, the Ala model for the cyclopeptide gives the clearest representation of the amide VCD. The experimental sign pattern for the amide I' band in D(2)O and also the sharper, more intense amide I VCD band in TFE was seen to some degree in one conformer with Type II' turns, but the data favor a mix of structures. PMID- 18506833 TI - Use of large-scale chromatography in the preparation of armodafinil. AB - Armodafinil, the (R)-enantiomer of modafinil, is a medication used to treat the excessive sleepiness associated with narcolepsy, obstructive sleep apnea/hypopnea syndrome, and shift work sleep disorder. We report here the chemical development of armodafinil and the investigations that led to a commercial route to prepare this pure enantiomer. Three synthetic approaches were used to provide the chiral sulfoxide. Resolution via preferential crystallization was used for phase I clinical trials and was subsequently replaced by chiral chromatography, enabling us to pursue a rapid filing and registration of the API. Finally, the commercial route was developed and employed asymmetric oxidation catalyzed by a titanium(IV) isopropoxide and diethyl tartrate system. The advantages of choosing a chromatographic development pathway to expedite registration while concurrently developing an economical chiral synthesis route is discussed in the context of armodafinil development. PMID- 18506834 TI - Chiral phosphites derived from carboranes: electronic effect in catalytic asymmetric hydrogenation. AB - A series of new fine-tunable monodentate phosphite and phosphoramidite ligands based on carboranes have been synthesized and used for asymmetric Rh-catalyzed hydrogenation of prochiral olefins with the result of up to 99.8% ee. Dependence of the enantioselectivity on the electron-withdrawing or electron-donating properties of the carboranyl substituent has been studied. PMID- 18506835 TI - A new reference material for UV-visible circular dichroism spectroscopy. AB - To obtain accurate and consistent measurements from circular dichroism (CD) instruments over time and from different laboratories, it is important that they are properly calibrated. The characteristics of the available reference materials are not ideal to ensure proper calibration as they typically only give peaks in one or two spectral regions, and often have issues concerning purity and stability. Currently either camphor sulfonic acid or ammonium camphor sulfonate are used. The latter can be an unstable, slightly hygroscopic secondary standard compound with only one characterized CD band. The former is the very hygroscopic primary standard for which only one enantiomer is readily available. We have synthesized a new reference material for CD, Na[Co(EDDS)].H(2)O (EDDS = N,N ethylenediaminedisuccinic acid) which addresses these problems. It is extremely stable and available in both enantiomeric forms. The CD spectrum of Na[Co(EDDS)].H(2)O has nine distinct peaks between 180 and 599 nm. It thus fulfils the principal requirements for CD calibration chemical standards and has the potential to be used to ensure good practice in the measurement of CD data, providing two spectra of equal magnitude and opposite sign for a given concentration and path length. We have carried out an interlaboratory comparison using this material and show how it can be used to improve CD comparability between laboratories. A fitting algorithm has been developed to assess CD spectropolarimeter performance between 750 and 178 nm. This could be the basis of a formal quality control process once criteria for performance have been decided. PMID- 18506836 TI - Highly enantioselective organocatalytic conjugate addition of nitromethane to benzylidene acetones. AB - Six active 4-aryl-5-nitro-pentan-2-ones were synthesized enantioselectively from the corresponding 5-aryl-butenones by asymmetric Michael addition of nitromethane using an imidazolidine-type enantioselective organocatalyst. The ee ratio of the products were between 67 and 100%, determined by HPLC with Chiracel OD. Molecular and crystal structure of 3,4-methylenedioxy-phenyl-5-nitro-pentan-2-one has been studied by single crystal X-ray diffraction. PMID- 18506837 TI - Diamagnetic lanthanide tris beta-diketonates as organic-soluble chiral NMR shift reagents. AB - Diamagnetic lanthanium(III) and lutetium(III) tris beta-diketonate complexes of 3 (trifluoroacetyl)-d-camphor, 3-(heptafluorobutyryl)-d-camphor, and d,d dicampholylmethane are shown to be effective chiral NMR shift reagents for determining the enantiomeric purity of compounds with hard Lewis base functional groups. These include substrates with amine, alcohol, epoxide, sulfoxide, and oxaxolidine moieties. Enantiomeric discrimination is observed in the (1)H NMR spectrum. Diamagnetic lanthanide complexes represent an alternative to paramagnetic varieties that often cause too much line broadening in the NMR spectra. The choice of which metal to use varies with substrate. Similarly, there is no consistent trend with ligand as not one of the complexes is consistently better than the others for all substrates. The enantiomeric discrimination also varies with solvent. Comparisons show that the chiral recognition was usually larger in benzene-d(6) than in chloroform-d or cyclohexane-d(12). PMID- 18506838 TI - Molecular dynamics study on the conformational flexibility and energetics in aqueous solution of methylated beta-cyclodextrins. AB - All possible methylated beta-cyclodextrins (CDs) with C7-symmetry have been studied by molecular dynamics simulations, in gas phase and in water solution. Energetic and structural information were obtained from the trajectory analysis. CD flexibility increases with degree of methylation, very likely due to the concomitant reduction of the intramolecular hydrogen bonds. Solvation-free energy was computed for each of the studied CDs using the MM/GBSA method. An analysis of radial distribution functions was used to determine distribution of solvent molecules around the O2, O3, and O6. The number of solvent molecules around these oxygens decreases with an increase in the degree of methylation. The DeltaS contribution from solvent thus becomes more positive when the degree of methylation increases and, consequently, the overall DeltaG in water diminishes. PMID- 18506839 TI - Liquid chromatographic direct resolution of tocainide and its analogs on a (3,3' diphenyl-1,1'-binaphthyl)-20-crown-6-based chiral stationary phase containing residual silanol protecting n-octyl groups. AB - Optically active (3,3'-diphenyl-1,1'-binaphthyl)-20-crown-6-based chiral stationary phase (CSP) containing residual silanol protecting n-octyl groups on silica surface was applied to the liquid chromatographic direct resolution of tocainide and its analogs. The chiral recognition ability of the CSP was excellent, the separation (alpha) and the resolution factors (R(S)) for 15 analytes including tocainide being in the range of 3.02-22.92 and 3.94-20.41, respectively. In addition, the chiral recognition ability of the CSP was much greater than that of (3,3'-diphenyl-1,1'-binaphthyl)-20-crown-6-based CSP containing residual silanol groups on the silica surface. The chromatographic behaviors for the resolution of tocainide and its analogs were found to be dependent on the content and the type of organic and acidic modifiers and the ammonium acetate concentration in aqueous mobile phase. PMID- 18506840 TI - Metabolic regulation by lactate. AB - For more than a century, the metabolic role of lactate has intrigued physiologists and biochemists. Yet, for the first half of the last century lactate had been designated as a waste product, and assigned no additional significance besides its controversial role in muscle fatigue. The decline of the lactate hypothesis for the onset of muscle fatigue and the defining of some modulatory properties attributed to lactate have increased the interest on this molecule. The present critical review aimed at evaluating some recent publications concerned with unveiling the regulatory actions of lactate in cellular function. Lactate has been described to modulate enzymes catalytic properties to affect hormonal release and responsiveness, and to control body homeostasis. Moreover, these properties are directly related to the genesis and the sustainability of pathological conditions, such as diabetes and cancer. In the end, we concluded that lactate should not be regarded as simply an anaerobic metabolite, but should be considered as a regulatory molecule that modulates the integration of metabolism. PMID- 18506841 TI - Ammonia induces RNA oxidation in cultured astrocytes and brain in vivo. AB - Oxidative stress plays a major role in cerebral ammonia toxicity and the pathogenesis of hepatic encephalopathy (HE). As shown in this study, ammonia induces a rapid RNA oxidation in cultured rat astrocytes, vital mouse brain slices, and rat brain in vivo. Ammonia-induced RNA oxidation in cultured astrocytes is reversible and sensitive to MK-801, 1,2-Bis(o-aminophenoxy)ethane N,N,N',N'-tetraacetic acid, apocynin, epigallocatechin gallate, and polyphenon 60, suggesting the involvement of N-methyl-D-aspartic acid (NMDA) receptor activation, Ca(2+), nicotinamide adenine dinucleotide phosphate, and reduced form (NADPH) oxidase-dependent oxidative stress. Also, hypo-osmolarity, tumor necrosis factor alpha (TNF-alpha), and diazepam increase RNA oxidation in cultured astrocytes, suggesting that the action of different HE-precipitating factors converges at the level of RNA oxidation. Among the oxidized RNA species, 18S-rRNA and the messenger RNA (mRNA) coding for the glutamate/aspartate transporter (GLAST) were identified. Cerebral RNA oxidation in acutely ammonia-loaded rats in vivo is reversible and predominates in neuronal soma and perivascular astrocyte processes. In neuronal dendrites, oxidized RNA colocalizes with the RNA-binding splicing protein neurooncological ventral antigen (NOVA)-2 within putative RNA transport granules, which are also found in close vicinity to postsynaptic spines. This indicates that oxidized RNA species may participate in postsynaptic protein synthesis, which is a biochemical substrate for learning and memory consolidation. Neuronal and astroglial RNA oxidation increases also in vital mouse brain slices treated with ammonia and TNF-alpha, respectively. CONCLUSION: Cerebral RNA oxidation is identified as a not yet recognized consequence of acute ammonia intoxication. RNA oxidation may affect gene expression and local protein synthesis and thereby provide another link between reactive oxygen species (ROS)/reactive nitrogen oxide species (RNOS) production and ammonia toxicity. PMID- 18506842 TI - Retinol-binding protein 4: a new marker of virus-induced steatosis in patients infected with hepatitis c virus genotype 1. AB - Retinol-binding protein 4 (RBP4) is an adipocytokine associated with insulin resistance (IR). We tested serum levels of RBP4 to assess its link with steatosis in patients with genotype 1 chronic hepatitis C (CHC) or nonalcoholic fatty liver disease (NAFLD). Nondiabetic patients with CHC (n = 143) or NAFLD (n = 37) were evaluated by liver biopsy and anthropometric and metabolic measurements, including IR by the homeostasis model assessment. Biopsies were scored by Scheuer classification for CHC, and Kleiner for NAFLD. Steatosis was tested as a continuous variable and graded as absent-mild <30%, or moderate-severe > or =30%. Thirty nondiabetic, nonobese blood donors served as controls. RBP4 levels were measured by a human competitive enzyme-linked immunosorbent assay kit (AdipoGen). Mean values of RBP4 were similar in NAFLD and CHC (35.3 +/- 9.3 microg/L versus 36.8 +/- 17.6; P = 0.47, respectively), and both were significantly higher than in controls (28.9 +/- 12.1; P = 0.02 and P = 0.01, respectively). RBP4 was higher in CHC patients with steatosis than in NAFLD (42.1 +/- 19.7 versus 35.2 +/- 9.3; P = 0.04). By linear regression, RBP4 was independently linked to steatosis only (P = 0.008) in CHC, and to elevated body mass index (P = 0.01) and low grading (P = 0.04) in NAFLD. By linear regression, steatosis was independently linked to homeostasis model assessment score (P = 0.03) and high RBP4 (P = 0.003) in CHC. By logistic regression, RBP4 was the only variable independently associated with moderate-severe steatosis in CHC (odds ratio, 1.045; 95% confidence interval, 1.020 to 1.070; P = 0.0004), whereas waist circumference was associated with moderate-severe steatosis in NAFLD (odds ratio, 1.095; 95% confidence interval, 1.007 to 1.192; P = 0.03). CONCLUSION: In nondiabetic, nonobese patients with genotype 1 CHC, serum RBP4 levels might be the expression of a virus-linked pathway to steatosis, largely unrelated to IR. PMID- 18506843 TI - Toll-like receptors and adaptor molecules in liver disease: update. AB - Toll-like receptors (TLRs) are pattern recognition receptors that recognize pathogen-associated molecular patterns and signal through adaptor molecules, myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain containing adaptor protein (TIRAP), Toll/IL-1 receptor domain containing adaptor inducing interferon-beta (TRIF), and TRIF-related adaptor molecule (TRAM) to activate transcription factors, nuclear factor (NF)-kappaB, activator protein 1 (AP-1), and interferon regulatory factors (IRFs) leading to the initiation of innate immunity. This system promptly initiates host defenses against invading microorganisms. Endogenous TLR ligands such as the products from dying cells may also engage with TLRs as damage-associated molecular patterns. Although Kupffer cells are considered the primary cells to respond to pathogen associated molecular patterns in the liver, recent studies provide evidence of TLR signaling in hepatic nonimmune cell populations, including hepatocytes, biliary epithelial cells, endothelial cells, and hepatic stellate cells. This review highlights advances in TLR signaling in the liver, the role of TLRs in the individual hepatic cell populations, and the implication of TLR signaling in acute and chronic liver diseases. We further discuss recent advances regarding cytosolic pattern recognition receptors, RNA helicases that represents a new concept in chronic hepatitis C virus infection. PMID- 18506844 TI - Quantitation with QUEST of brain HRMAS-NMR signals: application to metabolic disorders in experimental epileptic seizures. AB - Quantitation of High Resolution Magic Angle Spinning (HRMAS) Nuclear Magnetic Resonance (NMR) signals enables establishing reference metabolite profiles of ex vivo tissues. Signals are often contaminated by a background signal originating mainly from macromolecules and lipids and by residual water which hampers proper quantitation. We show that automatic quantitation of HRMAS signals, even in the presence of a background, can be achieved by the semi-parametric algorithm QUEST based on prior knowledge of a metabolite basis-set. The latter was quantum mechanically simulated with NMR-SCOPE and requires accurate spin parameters. The region of interest of spectra is a small part of the full spectral bandwidth. Reducing the computation time inherent to the large number of data-points is possible by using ER-Filter in a preprocessing step. Through Monte-Carlo studies, we analyze the performances of quantitation without and with ER-Filtering. Applications of QUEST to quantitation of 1H ex vivo HRMAS-NMR data of mouse brains after intoxication with soman, are demonstrated. Metabolic profiles obtained during status epilepticus and later when neuronal lesions are installed, are established. Acetate, Alanine, Choline and gamma-amino-butyric acid concentrations increase in the piriform cortex during the initial status epilepticus, when seizures are maximum; Lactate and Glutamine concentrations increase while myo-Inositol and N-acetylaspartate concentrations decrease when neuronal lesions are clearly installed. PMID- 18506845 TI - Automatic quantification of local and global articular cartilage surface curvature: biomarkers for osteoarthritis? AB - The objective of this study was to quantitatively assess the surface curvature of the articular cartilage from low-field magnetic resonance imaging (MRI) data, and to investigate its role in populations with varying radiographic signs of osteoarthritis (OA), cross-sectionally and longitudinally. The curvature of the articular surface of the medial tibial compartment was estimated both on fine and coarse scales using two different automatic methods which are both developed from an automatic 3D segmentation algorithm. Cross-sectionally (n=288), the surface curvature for both the fine- and coarse-scale estimates were significantly higher in the OA population compared with the healthy population, with P<0.001 and P<<0.001, respectively. For the longitudinal study (n=245), there was a significant increase in fine-scale curvature for healthy and borderline OA populations (P<0.001), and in coarse-scale curvature for severe OA populations (P<0.05). Fine-scale curvature could predict progressors using the estimates of those healthy at baseline (P<0.001). The inter-scan precision was 2.2 and 6.5 (mean CV) for the fine- and coarse scale curvature measures, respectively. The results showed that quantitative curvature estimates from low-field MRI at different scales could potentially become biomarkers targeted at different stages of OA. PMID- 18506846 TI - Hypoxia-inducible factor 1 and cancer pathogenesis. AB - Hypoxia-inducible factor 1 (HIF-1) is a transcriptional activator that mediates adaptive responses to hypoxia. HIF-1 activity is increased in the majority of human cancers as a result of genetic alterations and intratumoral hypoxia. HIF-1 activates the transcription of genes that increase O(2) availability by stimulating angiogenesis or that reprogram cellular metabolism to adapt to reduced O(2) availability. Proof of principle studies in mouse models suggests that inhibition of HIF-1 activity may have therapeutic effects, especially in combination with other anticancer drugs. PMID- 18506847 TI - Stem cell antigen-1 localizes to lipid microdomains and associates with insulin degrading enzyme in skeletal myoblasts. AB - Stem cell antigen-1 (Sca-1, Ly6A/E) is a glycosylphosphotidylinositol-anchored protein that identifies many tissue progenitor cells. We originally identified Sca-1 as a marker of myogenic precursor cells and subsequently demonstrated that Sca-1 regulates proliferation of activated myoblasts, suggesting an important role for Sca-1 in skeletal muscle homeostasis. Beyond its functional role in regulating proliferation, however, little is known about the mechanism(s) that drive Sca-1-mediated events. We now report that lipid microdomain organization is essential for normal myogenic differentiation, and that Sca-1 constitutively localizes to these domains during myoblast proliferation and differentiation. We also demonstrate that Sca-1 associates with insulin degrading enzyme (IDE), a catalytic protein responsible for the cleavage of mitogenic peptides, in differentiating myoblasts. We show that chemical inhibition of IDE as well as RNAi knockdown of IDE mRNA recapitulates the phenotype of Sca-1 interference, that is, sustained myoblast proliferation and delayed myogenic differentiation. These findings identify the first signaling protein that physically and functionally associates with Sca-1 in myogenic precursor cells, and suggest a potential pathway for Sca-1-mediated signaling. Future efforts to manipulate this pathway may lead to new strategies for augmenting the myogenic proliferative response, and ultimately muscle repair. PMID- 18506848 TI - Sox9, a key transcription factor of bone morphogenetic protein-2-induced chondrogenesis, is activated through BMP pathway and a CCAAT box in the proximal promoter. AB - Mouse embryonic fibroblasts (MEFs) can be differentiated into fully functional chondrocytes in response to bone morphogenetic protein-2 (BMP-2). The expression of Sox9, a critical transcription factor for the multiple steps of chondrogenesis, has been reported to be upregulated during this process. But the molecular mechanisms by which BMP-2 promotes chondrogenesis still remain largely unknown. The aim of the present study was therefore to investigate the underlying mechanism. In the MEFs, BMP-2 efficiently induced Sox9 expression along with chondrogenic differentiation in a time- and dose-dependent manner. SB203580, a specific inhibitor for p38 pathway, blocked BMP-2-induced chondrogenic differentiation as well as Sox9 expression and its transactivation of downstream genes. Forced expression of Smad6, a natural antagonist for BMP/Smad pathway, only inhibited Sox9 protein function without rendering any effects on its mRNA expression. A CCAAT box was identified in Sox9 promoter as the cis-elements responsible for BMP-2 stimulation. This study provides insight into the mechanisms underlying BMP-2-regulated Sox9 expression and activity in MEFs, and suggests differential roles of BMP-2/p38 and BMP-2/Smad pathways in modulating the function of Sox9 during chondrogenesis. PMID- 18506849 TI - Possible determinants of rapid virological response suggesting shorter courses of combination therapy for hepatitis C virus genotype 2. PMID- 18506850 TI - Adenosine-dependent activation of hypoxia-inducible factor-1 induces late preconditioning in liver cells. AB - The cellular mechanisms by which ischemic preconditioning increases liver tolerance to ischemia/reperfusion injury are still poorly understood. This study investigated the role of the hypoxia-inducible factor-1 (HIF-1) in the protection associated with the late phase of liver preconditioning. Late preconditioning was induced in primary cultured rat hepatocytes by a transient (10 minute) hypoxic stress or by 15 minutes incubation with the adenosine A(2A) receptors agonist CGS21680 24 hours before exposure to 90 minutes of hypoxia in a serum-free medium. Late preconditioning induced the nuclear translocation of HIF-1 and the expression of carbonic anhydrase IX (CAIX), a HIF-1-regulated transmembrane enzyme that catalyzes bicarbonate production. Such effects were associated with prevention of hepatocyte killing by hypoxia and the amelioration of intracellular acidosis and Na+ accumulation. The inhibition of PKC-mediated and PI3-kinase mediated signals with, respectively, chelerythrine and wortmannin abolished HIF-1 activation and blocked both CAIX expression and the protective action of late preconditioning. CAIX expression was also prevented by interfering with the transcriptional activity of HIF-1 using a dominant negative HIF-1beta subunit. The inhibition of CAIX with acetazolamide or the block of bicarbonate influx with disodium-4-acetamido-4'-isothiocyanato-stilben-2,2'-disulfonate also reverted the protective effects of late preconditioning on intracellular acidosis and Na+ accumulation. CONCLUSION: The stimulation of adenosine A(2A) receptors induced late preconditioning in liver cells through the activation of HIF-1. HIF-1 induced expression of CAIX increases hepatocyte tolerance to ischemia by maintaining intracellular Na+ homeostasis. These observations along with the importance of HIF-1 in regulating cell survival indicates HIF-1 activation as a possible key event in liver protection by late preconditioning. PMID- 18506851 TI - Posttreatment with the Ca(2+)-Mg(2+)-dependent endonuclease inhibitor aurintricarboxylic acid abolishes genotoxic agent-induced nuclear condensation and DNA fragmentation and decreases death of astrocytes. AB - DNA fragmentation and nuclear condensation are important nuclear changes in apoptosis. In this study we determined whether DNA fragmentation and nuclear condensation occur in astrocytes treated with 100-200 microM of the genotoxic agent M-nitroso-N-nitroguanidine (MNNG). Our study also investigated the roles of Ca(2+)-Mg(2+)-dependent endonuclease (CME) in the MNNG-induced nuclear changes. We found that MNNG induced profound ATP depletion as well as marked nuclear condensation and DNA fragmentation in the cells. Both the nuclear condensation and the DNA fragmentation were abolished by posttreatment of the cells with the CME inhibitor aurintricarboxylic acid (ATA). The ATA posttreatment also significantly, but only partially, decreased MNNG-induced cell death. In contrast, pretreatment plus cotreatment with ATA did not affect either MNNG induced nuclear condensation or cell death. Our study further suggests that ATA does not decrease the cytotoxicity of MNNG by directly inhibiting poly(ADP ribose) polymerases. Collectively, our observations suggest that MNNG can induce both DNA fragmentation and nuclear condensation in astrocytes by a CME-dependent mechanism, which partially contributes to the genotoxic agent-induced cell death. Published 2008 Wiley-Liss, Inc. PMID- 18506852 TI - Gene expression patterns in mouse cortical penumbra after focal ischemic brain injury and reperfusion. AB - Ischemic stress in the brain causes acute and massive cell death in the targeted core area followed by a second phase of damage in the neighboring penumbra. The purpose of this study was to examine the global gene expression patterns in the penumbra, because the ischemic lesion in this region could be rescued by restoration of blood flow and other protective therapies. Adult C57Bl/6 mice were subjected to a 90-min middle cerebral artery occlusion (MCAO). Laser capture microdissection (LCM) was used for tissue dissection at 4 and 24 hr after reperfusion. Sham-operated animals were used as controls. Gene expression in the penumbra was examined by using microarray analysis and quantitative RT-PCR. In agreement with previous reports, most genes were down-regulated at 4 hr after the onset of reperfusion in the ischemic penumbra compared with controls. In contrast, at 24 hr after reperfusion, most genes were up-regulated in the ischemic penumbra. Several genes not previously reported to be associated with ischemia were found. The gene lists generated in this study will help us to understand better the spatial and temporal distribution of molecules involved in the ischemic cascade. Published 2008 Wiley-Liss, Inc. PMID- 18506853 TI - Kaempferol: a new immunosuppressant of calcineurin. AB - Calcineurin (CN), the Ca(2+)/calmodulin (CaM)-dependant protein phosphatase, is the target for immunosuppressive drugs cyclosporine A (CsA) and FK506. These immunosuppressants can inhibit CN activity after binding with respective immunophilins. Based on the model of screening by using p-nitrophenyl phosphate as a substrate for preliminary screening and (32)P-labeled 19-residue phosphopeptide as a specific substrate for final determination, we found Kaempferol, a natural flavonol, could inhibit CN activity in purified enzyme and Jurkat T-cells. Unlike CsA and FK506, CN inhibition by kaempferol is independent of matchmaker protein and the inhibitory manner is noncompetitive. Through investigation of inhibitions for CNA and a series of its truncated mutants, we suggested that Kaempferol could directly act on the catalytic domain. Data also indicated that the CN inhibition by kaempferol could be enhanced when the enzyme was activated in the presence of CaM and CNB. CNB is necessary for mediating inhibition of enzyme by kaempferol. The result of RT-PCR also indicated that kaempferol had an inhibitory activity against IL-2 gene expression in activated Jurkat cells. All data suggested that kaempferol could be a new immunosuppressant of CN. PMID- 18506854 TI - Proteasome activation as a novel antiaging strategy. AB - Homeostasis is a key feature of cellular lifespan. Maintenance of cellular homeostasis influences the rate of aging and is determined by several factors, including efficient proteolysis of damaged proteins. Protein degradation is predominantly catalyzed by the proteasome. Specifically, the proteasome is responsible for cell clearance of abnormal, denatured or in general damaged proteins as well as for the regulated degradation of short-lived proteins. As proteasome has an impaired function during aging, emphasis has been given recently in identifying ways of its activation. A number of studies have shown that the proteasome can be activated by genetic manipulations as well as by factors that affect its conformation and stability. Importantly the developed proteasome activated cell lines exhibit an extended lifespan. This review article discusses in details the various factors that are involved in proteasome biosynthesis and assembly and how they contribute to its activation. Finally as few natural compounds have been identified having proteasome activation properties, we discuss the advantages of this novel antiaging strategy. PMID- 18506856 TI - Management of gastric fundal varices. PMID- 18506855 TI - Physical chemistry of intestinal absorption of biliary cholesterol in mice. AB - Although many putative sterol transporters influencing cholesterol absorption and physical-chemical factors affecting dietary cholesterol absorption have been extensively investigated, it is still unclear how biliary cholesterol contributes to the regulation of intestinal cholesterol absorption. We studied whether the gallbladder can modulate the microaggregates of cholesterol carriers, which may in turn influence the intestinal absorption of biliary cholesterol. Supersaturated, crystallized, or micellar model biles were delivered via a duodenal catheter to conscious, freely moving C57L mice daily for 2 days. Intestinal uptake and absorption of biliary cholesterol and its fecal excretion, as well as expression levels of intestinal sterol transporters, were analyzed. Cholesterol uptake and absorption by the enterocyte were dramatically reduced in mice treated with crystallized biles compared with supersaturated biles. This correlated with the higher cumulative radioactivity of cholesterol recovered in the feces at 24 hours. Such findings were absent with the added reference compound sitostanol. After removing cholesterol crystals from crystallized biles, micellar biles showed essentially identical effects on intestinal absorption but with lower fecal cholesterol excretion compared with the original samples containing crystals. Expression levels of the jejunal Abcg5 (ATP-binding cassette transporter G5) and Abcg8, but not Npc1l1 (Niemann-Pick C1 like 1), were significantly increased by supersaturated biles compared with crystallized biles. CONCLUSION: Different physical forms of biliary cholesterol dramatically determine intestinal uptake and absorption of cholesterol. Solid plate-like cholesterol monohydrate crystals in bile are probably not absorbed and are totally excreted in feces from the body. The gallbladder may have a role in regulating cholesterol homeostasis by modulating the physical forms of biliary cholesterol. PMID- 18506857 TI - Photoinduced luminescence blinking and bleaching in individual single-walled carbon nanotubes. AB - The temporal evolution of photoluminescence in individual single-walled carbon nanotubes (SWNT) under strong laser irradiation is studied and pronounced blinking and bleaching is observed, caused by photoinduced oxidation that subsequently quenches mobile excitons. The nanotubes are isolated with sodium cholate and spun onto either a glass or mica surface. Their bleaching behavior is investigated for variable laser intensities in air and argon atmosphere. The decay rate for luminescence bleaching generally increases with higher laser intensity, however saturating on mica substrates, which is attributed to limited availability of oxygen in the vicinity of the nanotubes. Step-like events in the luminescence time traces corresponding to single oxidation events are analyzed regarding relative step height and suggest an exciton diffusion range of about 105 nm. PMID- 18506858 TI - Azacalix[6]arene hexamethyl ether: synthesis, structure, and selective uptake of carbon dioxide in the solid state. AB - To investigate dynamic solid-state complexation hitherto unexplored in nitrogen bridged calixarene analogues, azacalix[6]arene hexamethyl ether has been prepared in three steps by applying a 5+1 fragment-coupling approach by using a Buchwald- Hartwig aryl amination reaction with the aid of our previously devised temporal N silylation protocol. X-ray crystallographic analysis and NMR spectroscopic measurements have revealed that the azacalix[6]arene is well endowed with hydrogen-bonding ability, by which both the molecular and crystal structures are controlled. The azacalix[6]arene is conformationally flexible in solution on the NMR time scale, whereas it adopts a definite 1,2,3-alternate conformation with S2 symmetry in the solid state as a result of intramolecular bifurcated hydrogen bonding interactions. In the crystal, molecules of the azacalix[6]arene are mutually interacted by intermolecular hydrogen bonds to establish one-dimensional hexane-filled nanochannel crystal architecture. Although the single crystal was broken after desolvation, the resultant polycrystalline powder material was capable of selectively adsorbing CO2 among the four main gaseous components of the atmosphere. In contrast, carbocyclic p-tert-butylcalix[6]arene hexamethyl ether, the crystal structure of which was also elucidated for the first time in the present study, gave rise to almost no uptake of CO2. Additional solid-gas adsorption experiments for another three gases, such as N2, O2, and Ar, suggested that quadrupole/induced-dipole interactions and/or hydrogen-bonding interactions played an important role in permitting the observed selective uptake of CO2 by this new azacalix[6]arene in the solid state. PMID- 18506859 TI - Classical reagents: new surprises in palladium-catalyzed C--C coupling reactions. AB - From investigations on a relatively simple concept to identify conditions for promoting Mizoroki-Heck reactions with vinyl tosylates and phosphates, two serendipitous discoveries were made concerning new properties of palladium as a catalyst. In the first case, beta-hydride eliminations well known for alkyl metal complexes were found to be equally feasible with alkenyl metal compounds. And secondly, conditions were found for promoting intermolecular ene-yne couplings via a PdII-H intermediate. This coupling reactions represents an atom economical Mizoroki-Heck type reaction. PMID- 18506860 TI - Enhanced pi conjugation around a porphyrin[6] nanoring. PMID- 18506861 TI - Efficient helicene synthesis: Friedel-Crafts-type cyclization of 1,1-difluoro-1 alkenes. PMID- 18506862 TI - Iron-catalyzed Sonogashira reactions. PMID- 18506863 TI - Synthesis of a stabilized version of the imidazolone DNA lesion. AB - Imidazolone (dIz) is an abundant, highly mutagenic, and rather unstable DNA lesion that can cause dG-->dC transversion mutations. dIz is generated in DNA by a variety of oxidative processes such as type I photooxidation. Herein we report the synthesis of a carbocyclic nucleoside analogue of dIz and of DNA containing this stabilized lesion analogue. The carbocyclic modification protects this lesion analogue from anomerization. As the repair of the lesion analogue by DNA glycosylases is not possible, this analogue should allow cocrystallization studies together with wild-type repair enzymes. Characterization of the lesion analogue was performed by using spectroscopic methods and enzymatic digestion experiments of the oligonucleotides. PMID- 18506864 TI - Isomerization of the Asp7 residue results in zinc-induced oligomerization of Alzheimer's disease amyloid beta(1-16) peptide. PMID- 18506865 TI - Heparin dependent coiled-coil formation. PMID- 18506866 TI - Gas phase ion chemistry of gold-silicon clusters. PMID- 18506867 TI - Organic-inorganic hybrid materials for efficient enantioseparation using cellulose 3,5-dimethylphenylcarbamate and tetraethyl orthosilicate. AB - The hybrid bead-type chiral packing material (CPM) for preparative enantioseparation has been prepared from the cellulose 3,5 dimethylphenylcarbamate containing a small number of 3-(triethoxysilyl)propyl groups in the presence of tetraethyl orthosilicate, by a sol-gel reaction in an aqueous surfactant solution. The obtained hybrid bead-type CPM was packed into a column and evaluated by high-performance liquid chromatography. When compared with the commercially available Chiralpak IB, which is prepared by the immobilization of cellulose 3,5-dimethylphenylcarbamate on silica gel, the hybrid bead-type CPM was shown to exhibit a similar chiral recognition and possess a higher loading capacity. PMID- 18506868 TI - NMR studies on monofunctionalized fullerenyl cation and anion encapsulating a H2 molecule. AB - (1)H NMR chemical shifts of molecular hydrogen encapsulated in dichloromethyl C(60) cation and (1-octynyl)-C(60) anion were studied to clarify the difference in magnetic shielding effects inside the fullerene cages. The signals of the H(2) molecule inside both cationic and anionic C(60) cages appeared at lower fields than those of the neutral counterparts. These results were interpreted based on the results of NICS calculations. It was demonstrated that the H(2) molecule inside the C(60) cages can serve as an excellent NMR probe of aromaticity for both cationic and anionic C(60) derivatives. PMID- 18506869 TI - The molecular mechanism of tropospheric nitrous acid production on mineral dust surfaces. PMID- 18506870 TI - Self-assembly of two different hierarchical nanostructures on either side of an organic supramolecular film in one step. AB - We fabricated different hierarchical organic nanostructures on each side of a supramolecular film, by using hydrogen-bonding interactions between tetrapyridylporphyrin and benzene-1,3,5-tricarboxylic acid at the H2O/CHCl3 interface. The surface of the film that faces water is composed of nanoprism arrays, whereas the surface facing CHCl3 is composed of three-dimensional sunflower-like hierarchical micro- and nanostructures. FTIR spectral evidence showed that all pyridyl groups of the tetrapyridylporphyrin hydrogen bonded to the carboxylic acid groups of 1,3,5-benzene-tricarboxylic acid. The aggregation modes of porphyrin presented in this supramolecular film were studied by UV/Vis and fluorescence spectroscopy. Moreover, each side of the film exhibits distinct soakage properties. PMID- 18506871 TI - The Rehm-Weller experiment in view of distant electron transfer. AB - The driving-force dependence of bimolecular fluorescence quenching by electron transfer in solution, the Rehm-Weller experiment, is revisited. One of the three long-standing unsolved questions about the features of this experiment is carefully analysed here, that is, is there a diffusional plateau? New experimental quenching rates are compiled for a single electron donor, 2,5 bis(dimethylamino)-1,3-benzenedicarbonitrile, and eighteen electron acceptors in acetonitrile. The data are analysed in the framework of differential encounter theory by using an extended version of the Marcus theory to model the intrinsic electron-transfer step. Only by including the hydrodynamic effect and the solvent structure can the experimental findings be well modelled. The diffusional control region, the "plateau", reveals the inherent distance dependence of the reaction, which is shown to be a general feature of electron transfer in solution. PMID- 18506873 TI - Glow-discharge plasma-assisted design of cobalt catalysts for Fischer-Tropsch synthesis. PMID- 18506872 TI - Monolayer-barcoded nanoparticles for on-chip DNA hybridization assay. PMID- 18506874 TI - Two functionally redundant Sfp-type 4'-phosphopantetheinyl transferases differentially activate biosynthetic pathways in Myxococcus xanthus. PMID- 18506875 TI - The corrin moiety of coenzyme B12 is the determinant for switching the btuB riboswitch of E. coli. AB - Riboswitches are regulatory elements in the 5'-untranslated region (5'-UTR) of bacterial mRNAs that bind certain metabolites with high specificity and affinity. The 202 nucleotide (nt)-long btuB riboswitch RNA of E. coli interacts specifically with coenzyme B12 and its derivatives thereby leading to changes in the RNA structure and hence to an altered expression of the downstream btuB gene. We report the investigations of the rearrangement of the three-dimensional structure of the btuB riboswitch upon binding to four different B12 derivatives: coenzyme B12, vitamin B12, adenosyl factor A and adenosyl-cobinamide. In-line probing experiments have shown that the corrin ring plays the crucial role in switching the three-dimensional riboswitch structure. Instead, the apical ligands influence only the binding affinity of the B12 derivative to the btuB riboswitch. PMID- 18506876 TI - A chemical library approach to organic-modified peptide ligands for PDZ domain proteins: a synthetic, thermodynamic and structural investigation. PMID- 18506877 TI - MicroRNA expression profiling: a molecular tool for defining the phenotype of hepatocellular tumors. PMID- 18506878 TI - Location is everything: the liver stem cell niche. PMID- 18506879 TI - CD4+ T cell help improves CD8+ T cell memory by retained CD27 expression. AB - CD4+ T cell help during the priming of CD8+ T lymphocytes imprints the capacity for optimal secondary expansion upon re-encounter with antigen. Helped memory CD8+ T cells rapidly expand in response to a secondary antigen exposure, even in the absence of T cell help and, are most efficient in protection against a re infection. In contrast, helpless memory CTL can mediate effector function, but secondary expansion is reduced. How CD4+ T cells instruct CD8+ memory T cells during priming to undergo efficient secondary expansion has not been resolved in detail. Here, we show that memory CTL after infection with lymphocytic choriomeningitis virus are CD27(high) whereas memory CTL primed in the absence of CD4+ T cell have a reduced expression of CD27. Helpless memory CTL produced low amounts of IL-2 and did not efficiently expand after restimulation with peptide in vitro. Blocking experiments with monoclonal antibodies and the use of CD27(-/ ) memory CTL revealed that CD27 ligation during restimulation increased autocrine IL-2 production and secondary expansion. Therefore, regulating CD27 expression on memory CTL is a novel mechanism how CD4+ T cells control CTL memory. PMID- 18506880 TI - Generating functional CD8+ T cell memory response under transient CD4+ T cell deficiency: implications for vaccination of immunocompromised individuals. AB - Studies based on either MHC class II-knockout or CD4+ T cell-depleted murine models have demonstrated a critical role for CD4+ T cells in the generation of CD8+ T cell memory. However, it is difficult to extend these findings to immunocompromised humans where a complete loss of CD4+ T cells is rarely observed. Here, we have developed a model setting, which allows studies on the generation of CD8+ T cell memory responses in a transient CD4+ T cell-deficient setting similar to that seen in immunocompromised patients. Immunisation with an adenoviral vaccine under transient helpless or help-deficient conditions showed varying degrees of impact on the priming of CD8+ T cell responses. Antigen specific T cells generated under normal CD4+ T cell help and transient help deficient conditions showed similar effector phenotype and were capable of proliferation upon secondary antigen encounter. Most importantly, in spite of CD4+ T cell deficiency, the long-term CD8+ T cell memory response remained functionally stable and showed comparable cytotoxic effector function as seen in CD8+ T cells generated with normal CD4+ T cell numbers. These findings provide evidence that in spite of partially impaired activation of a primary CD8+ T cell response, a fully functional and stable memory CTL response can be induced under conditions of severe transient CD4+ T cell deficiency. PMID- 18506881 TI - HLA-DM negatively regulates HLA-DR4-restricted collagen pathogenic peptide presentation and T cell recognition. AB - Rheumatoid arthritis, an autoimmune disease, is significantly associated with the HLA class II allele HLA-DR4. While the etiology of rheumatoid arthritis remains unknown, type II collagen (CII) is a candidate autoantigen. An immunodominant pathogenic epitope from this autoantigen, CII(261-273), which binds to HLA-DR4 and activates CD4+ T cells, has been identified. The non-classical class II antigen, HLA-DM, is also a key component of class II antigen presentation pathways influencing peptide presentation by HLA-DR molecules expressed on professional antigen-presenting cells (APC). Here, we investigated whether the HLA-DR4-restricted presentation of the pathogenic CII(261-273) epitope was regulated by HLA-DM expression in APC. We show that APC lacking HLA-DM efficiently display the CII(261-273) peptide/epitope to activate CD4+ T cells, and that presentation of this peptide is modulated dependent on the level of HLA DM expression in APC. Mechanistic studies demonstrated that the CII(261-273) peptide is internalized by APC and edited by HLA-DM molecules in the recycling pathway, inhibiting peptide presentation and T cell recognition. These findings suggest that HLA-DM expression in APC controls class II-mediated CII(261-273) peptide/epitope presentation and regulates CD4+ T cell responses to this self epitope, thus potentially influencing CII-dependent autoimmunity. PMID- 18506883 TI - Blood CD8+ T cell responses against myelin determinants in multiple sclerosis and healthy individuals. AB - Patients with multiple sclerosis (MS) display significant peripheral blood CD8(+) T cell receptor biases, suggesting clonal selection. Our objective was to identify relevant myelin-derived peptides capable of eliciting responses of fresh blood CD8+ T cells in MS patients. We focused our analysis on the HLA supertypes (HLA-A3, -A2, -B7, -B27, -B44) predominant in a patient cohort. Three myelin protein (MBP, PLP and MOG) sequences were screened for HLA binding motifs and peptides were tested for their binding to HLA molecules. The cellular responses of 27 MS patients and 19 age- and sex-matched healthy controls (HC) were tested in IFN-gamma ELISPOT assays only detecting pre-committed CD8+ T cells. Sixty-nine new epitopes elicited positive responses, with MOG-derived peptides being the most immunogenic and peptides binding to HLA-A3 being the most frequent. However, MS patients and HC displayed the same frequency of autoreactive cells. The epitopes inducing the strongest responses were not those with the highest HLA binding, suggesting an effective thymic selection in MS patients. Our data extend the concept that the frequency of myelin-reactive T cells in MS patient blood is not increased compared to HC. The description of this set of myelin-derived peptides (MHC class I restricted, recognized by CD8+ T cells) offers new tools to explore the CD8+ cell role in MS. PMID- 18506882 TI - Systemic IFN-alpha drives kidney nephritis in B6.Sle123 mice. AB - The impact of IFN-alpha secretion on disease progression was assessed by comparing phenotypic changes in the lupus-prone B6.Sle1Sle2Sle3 (B6.Sle123) strain and the parental C57BL/6 (B6) congenic partner using an adenovirus (ADV) expression vector containing a recombinant IFN-alpha gene cassette (IFN-ADV). A comprehensive comparison of cell lineage composition and activation in young B6 and B6.Sle123 mice revealed a variety of cellular alterations in the presence and absence of systemic IFN-alpha. Most IFN-alpha-induced phenotypes were similar in B6 and B6.Sle123 mice; however, B6.Sle123 mice uniquely exhibited increased B1 and plasma cells after IFN-alpha exposure, although both strains had an overall loss of mature B cells in the bone marrow, spleen and periphery. Although most of the cellular effects of IFN-alpha were identical in both strains, severe glomerulonephritis occurred only in B6.Sle123 mice. Mice injected with IFN-ADV showed an increase in immune complex deposition in the kidney, together with an unexpected decrease in serum anti-nuclear antibody levels. In summary, the predominant impact of systemic IFN-alpha in this murine model is an exacerbation of mechanisms mediating end organ damage. PMID- 18506884 TI - Mutual antagonistic relationship between prostaglandin E(2) and IFN-gamma: Implications for rheumatoid arthritis. AB - Prostaglandin E(2) (PGE(2)) is a major mediator of inflammation and is present at high concentrations in the synovial fluid of rheumatoid arthritis (RA) patients. PGE(2), acting through the EP4 receptor, has both pro- and anti-inflammatory roles in vivo. To shed light on this dual role of PGE(2), we investigated its effects in whole blood and in primary human fibroblast-like synoviocytes (FLS). Gene expression analysis in human leukocytes, confirmed at the protein level, revealed an EP4-dependent inhibition of the expression of genes involved in the IFN-gamma-activation pathway, including IFN-gamma itself. This effect of the PGE(2)/EP4 axis on IFN-gamma is a reciprocal phenomenon since IFN-gamma blocks PGE(2) release and blocks EP receptor expression. The mutually antagonistic relationship between IFN-gamma and PGE(2) extends to downstream cytokine and chemokine release; PGE(2) counters the effects of IFN-gamma, on the release of IP 10, IL-8, TNF-alpha and IL-1beta. To gain further insight into IFN-gamma-mediated cellular events in RA, we assessed the effects of IFN-gamma on gene expression in FLS. We observed an IFN-gamma-dependent up-regulation of macrophage-attracting chemokines, and down-regulation of metalloprotease expression. These results suggest the existence of a mutually antagonistic relationship between PGE(2) and IFN-gamma, which may represent a fundamental mechanism of immune control in diseases such as RA. PMID- 18506885 TI - Interleukin 10 suppresses Th17 cytokines secreted by macrophages and T cells. AB - IL-17 and IL-22 are typical cytokines produced by the Th17 T cell subset, but it is unclear if Th17 cytokines can be produced by other cell types. We demonstrate that IL-10-deficient and IL-10R-deficient macrophages stimulated with lipopolysaccharide produce high levels of IL-17 and IL-22. Addition of exogenous IL-10 to IL-10-deficient macrophages abolished IL-17 production. When IL-10 deficient and IL-10R-deficient splenocytes were cultured under Th17 polarizing conditions, the population of IL-17-producing cells was increased and the cultures produced significantly higher levels of IL-17 and IL-22. The addition of recombinant IL-10 to IL-10-deficient splenocytes significantly decreased the percentage of IL-17-producing CD4(+) T cells. Finally, the mRNA for the Th17 transcription factor retinoic acid-related orphan receptor (ROR)gammat was significantly elevated in IL-10-deficient spleen cells and macrophages. These data demonstrate that Th17 cytokines and RORgammat are also expressed in macrophages and that IL-10 negatively regulates the expression of Th17 cytokines and RORgammat by both macrophages and T cells. PMID- 18506886 TI - Using RNA interference to identify the different roles of SMAD2 and SMAD3 in NIH/3T3 fibroblast cells. AB - Smad proteins are principal intracellular signaling mediators of transforming growth factor beta (TGF-beta) that regulate a wide range of biological processes. However, the identities of Smad partners mediating TGF-beta signaling are not fully understood. We firstly examined the expression of Smad2 and Smad3 induced by TGF-beta 1 in normal NIH/3T3 cells. The expression of Smad2 and Smad3 was assessed by RT-PCR and Western blotting. The results showed that the expression of Smad2 was increased after treatment with TGF-betaI, but Smad3 was more sensitive to TGF-betaI than Smad2. RNA interference (RNAi) provides a new approach for elucidation of gene function. Use of hairpin siRNA expression vectors for RNAi has provided a rapid and versatile method for assessing gene function in mammalian cells. Here, we have constructed Smad2 and Smad3 hairpin siRNA expression plasmids, and then transfected them into mouse NIH/3T3 cells. Endogenous Smad2 and Smad3 proteins decreased significantly at 48 h after transfection. We found the expression of Smad3 in Smad2-depleted cells was increased, however, the expression of Smad2 in Smad3-depleted cells was not changed. Consistently, the expression of Smad4 mRNA was also attenuated in Smad3 depleted cells. From these data, we suggest that Smad3, but not Smad2, may play a key role in TGF-beta signaling. PMID- 18506888 TI - Melittin prevents liver cancer cell metastasis through inhibition of the Rac1 dependent pathway. AB - Melittin, a water-soluble toxic peptide derived from bee venom of Apis mellifera was reported to have inhibitory effects on hepatocellular carcinoma (HCC). However, its role in antimetastasis and the underlying mechanism remains elusive. By utilizing both HCC cell lines and an animal model based assay system, we found that Rac1, which has been shown to be involved in cancer cell metastasis, is highly expressed in aggressive HCC cell lines and its activity correlated with cell motility and cytoskeleton polymerization. In addition, Rac1-dependent activity and metastatic potential of aggressive HCC cells are remarkably high in both cellular and nude mouse models. We provide evidence here that melittin inhibits the viability and motility of HCC cells in vitro, which correlates with its suppression of Rac1-dependent activity, cell motility, and microfilament depolymerization. Furthermore, melittin suppresses both HCC metastasis and Rac1 dependent activity in nude mouse models. The specificity of the effect of melittin on Rac1 was confirmed in HCC cells both in vitro and in vivo. CONCLUSION: Melittin inhibits tumor cell metastasis by reducing cell motility and migration via the suppression of Rac1-dependent pathway, suggesting that melittin is a potential therapeutic agent for HCC. PMID- 18506889 TI - Metron factor-1 prevents liver injury without promoting tumor growth and metastasis. AB - Hepatocyte growth factor (HGF) is the most powerful hepatotrophic factor identified so far. However, the ability of HGF to promote tumor cell "scattering" and invasion raises some concern about its therapeutic safety. We compared the therapeutic efficacy of HGF with that of Metron Factor-1 (MF-1), an engineered cytokine derived from HGF and the HGF-like factor macrophage stimulating protein (MSP), in mouse models of acute and chronic liver injury. At the same time, we tested the ability of HGF and MF-1 to promote tumor growth, angiogenesis, and invasion in several mouse models of cancer. We show that (1) MF-1 and HGF stimulate hepatocyte proliferation in vitro; (2) MF-1 and HGF protect primary hepatocytes against Fas-induced and drug-induced apoptosis; (3) HGF but not MF-1 induces scattering and matrigel invasion of carcinoma cell lines in vitro; (4) HGF but not MF-1 promotes migration and extracellular matrix invasion of endothelial cells in vitro; (5) MF-1 and HGF prevent CCl(4)-induced acute liver injury as measured by alanine aminotransferase (ALT) levels, histology, terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) analysis, and phospho-histone-3 immunostaining; (6) MF-1 and HGF attenuate liver fibrosis caused by chronic CCl(4) intoxication and promote regeneration as measured by Sirius red staining, alpha-smooth muscle actin immunostaining, and Ki-67 analysis; (7) HGF but not MF-1 promotes tumor growth, angiogenesis, and metastasis in a variety of xenograft models; (8) HGF but not MF-1 promotes intrahepatic dissemination of hepatocarcinoma cells injected orthotopically. CONCLUSION: These data suggest that MF-1 is as effective as HGF at preventing liver injury and at promoting hepatocyte regeneration, but therapeutically safer than HGF because it lacks proangiogenic and prometastatic activity. PMID- 18506890 TI - Lack of inducible nitric oxide synthase leads to increased hepatic apoptosis and decreased fibrosis in mice after chronic carbon tetrachloride administration. AB - The role of nitric oxide (NO) in liver injury and fibrosis is unclear. The purpose of this study was to determine whether inducible NO synthase deficiency (iNOS(-/-)) affects liver injury and fibrosis produced in mice by chronic carbon tetrachloride (CCl(4)) administration. Wild-type (WT) or iNOS(-/-) mice were subjected to biweekly CCl(4) injections over 8 weeks, whereas controls were given isovolumetric injections of olive oil. Serum aminotransferases were lower after CCl(4) in the iNOS(-/-) than in the WT mice, which correlated with decreased necrosis on liver histology. There was increased apoptosis, a lower number of stellate cells, and a lesser degree of fibrosis after CCl(4) in the iNOS(-/-) as compared with the WT mice. alpha(1)(I) collagen messenger RNA (mRNA) was markedly increased after CCl(4) in the WT and to a significantly lesser extent in the iNOS(-/-) mice. Liver matrix metalloproteinase-9 (MMP-9) mRNA and MMP-2 mRNA were increased more in the WT than in the iNOS(-/-) mice after CCl(4). Also tissue inhibitor metalloproteinase 1 (TIMP-1) mRNA was increased to a much greater extent in the WT than in the iNOS(-/-) mice after CCl(4) (P < 0.05). However, MMP 9 and TIMP-1 protein, determined by western blot, were similarly increased after CCl(4) in both groups of mice. CONCLUSION: NO protects against CCl(4)-induced apoptosis. In the absence of iNOS, there is decreased necrosis, increased apoptosis, and reduced liver fibrosis. PMID- 18506891 TI - Transforming growth factor-beta gene expression signature in mouse hepatocytes predicts clinical outcome in human cancer. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers in the world. The clinical heterogeneity of HCC, and the lack of good diagnostic markers and treatment strategies, has rendered the disease a major challenge. Patients with HCC have a highly variable clinical course, indicating that HCC comprises several biologically distinctive subgroups reflecting a molecular heterogeneity of the tumors. Transforming growth factor beta (TGF-beta) is known to exhibit tumor stage dependent suppressive (that is, growth inhibition) and oncogenic (that is, invasiveness) properties. Here, we asked if a TGF-beta specific gene expression signature could refine the classification and prognostic predictions for HCC patients. Applying a comparative functional genomics approach we demonstrated that a temporal TGF-beta gene expression signature established in mouse primary hepatocytes successfully discriminated distinct subgroups of HCC. The TGF-beta positive cluster included two novel homogeneous groups of HCC associated with early and late TGF-beta signatures. Kaplan-Meier plots and log-rank statistics indicated that the patients with a late TGF-beta signature showed significantly (P < 0.005) shortened mean survival time (16.2 +/- 5.3 months) compared to the patients with an early (60.7 +/- 16.1 months) TGF-beta signature. Also, tumors expressing late TGF-beta-responsive genes displayed invasive phenotype and increased tumor recurrence. We also showed that the late TGF-beta signature accurately predicted liver metastasis and discriminated HCC cell lines by degree of invasiveness. Finally, we established that the TGF-beta gene expression signature possessed a predictive value for tumors other than HCC. CONCLUSION: These data demonstrate the clinical significance of the genes embedded in TGF beta expression signature for the molecular classification of HCC. PMID- 18506892 TI - Capacitative calcium entry and transient receptor potential canonical 6 expression control human hepatoma cell proliferation. AB - Store-operated calcium entry (SOCE) is the main Ca(2+) influx pathway involved in controlling proliferation of the human hepatoma cell lines Huh-7 and HepG2. However, the molecular nature of the calcium channels involved in this process remains unknown. Huh-7 and HepG2 cells express transient receptor potential canonical 1 (TRPC1) and TRPC6, as well as STIM1 and Orai1, and these 4 channels are the most likely candidates to account for the SOCE in these cells. We generated stable TRPC6-overexpressing or TRPC6-knockdown Huh-7 clones, in which we investigated correlations between the presence of the protein, the rate of cell proliferation, and SOCE amplitude. TRPC6-overexpressing Huh-7 cells proliferated 80% faster than did untransfected cells and their SOCE amplitude was 160% higher. By contrast, proliferation rate was 50% lower and SOCE amplitude 85% lower in TRPC6-knockdown clones than in untransfected cells. OAG (olyl acetyl glycerol)-induced calcium entry was similar in all cells, and small interfering RNA (siRNA) against TRPC1 had no effect on SOCE amplitude, highlighting the relationship among SOCE, TRPC6 and cell proliferation in Huh-7 cells. SOCE amplitude was reduced by STIM1 and Orai1 knockdowns, suggesting possible cooperation between these proteins and TRPC6 in these cells. Endothelial growth factor and hepatocyte growth factor increased TRPC6 expression and SOCE amplitude in Huh-7 cells, and cyclin D1 expression was decreased by STIM1, Orai1, and TRPC6 knockdowns. CONCLUSION: TRPC6 was very weakly expressed in isolated hepatocytes from healthy patients and expressed more strongly in tumoral samples from the liver of a cancer patient, strongly supporting a role for these calcium channels in liver oncogenesis. PMID- 18506893 TI - Defective DNA strand break repair causes chromosomal instability and accelerates liver carcinogenesis in mice. AB - Chromosomal instability is a characteristic feature of hepatocellular carcinoma (HCC) but its origin and role in liver carcinogenesis are undefined. We tested whether a defect in the nonhomologous end-joining (NHEJ) DNA repair gene Ku70 was associated with chromosomal abnormalities and enhanced liver carcinogenesis. Male Ku70 NHEJ-deficient (Ku70-/-), heterozygote (Ku70 +/-), and wild-type (WT) mice were injected with diethylnitrosamine (DEN), a liver carcinogen, at age 15 days. Animals were killed at 3, 6, and 9 months for assessment of tumorigenesis and hepatocellular proliferation. For karyotype analysis, primary liver tumor cell cultures were prepared from HCCs arising in Ku70 mice of all genotypes. Compared to WT littermates, Ku70-/- mice injected with DEN displayed accelerated HCC development. Ku70-/- HCCs harbored clonal increases in numerical and structural aberrations of chromosomes 4, 5, 7, 8, 10, 14, and 19, many of which recapitulated the spectrum of equivalent chromosomal abnormalities observed in human HCC. Ku70-/- HCCs showed high proliferative activity with increased cyclin D1 and proliferating cell nuclear antigen expression, Aurora A kinase activity, enhanced ataxia telangiectasia mutated kinase and ubiquitination, and loss of p53 via proteasomal degradation, features which closely resemble those of human HCC. CONCLUSION: These findings demonstrate that defects in the NHEJ DNA repair pathway may participate in the disruption of cell cycle checkpoints leading to chromosomal instability and accelerated development of HCC. PMID- 18506894 TI - Diagnosis and treatment of Wilson disease: an update. PMID- 18506895 TI - Fphog1, a HOG-type MAP kinase gene, is involved in multistress response in Fusarium proliferatum. AB - Delta Fphog1 mutants of Fusarium proliferatum obtained by targeted gene disruption of Fphog1, an orthologue of the Saccharomyces cerevisiae hog1 MAPK gene showed increased sensitivity towards different abiotic stressors including UV-irradiation, heat, salt, osmotic and hydrogen peroxide treatments. Incubation of the Delta Fphog1 mutants under hyperosmotic conditions was accompanied with prolonged growth arrest, inhibition of conidial germination, morphological abnormalities and time-dependent increase of the cell death rate. The wild type Fphog1 gene, under the control of its own promoter, was able to rescue the multistress sensitivity of the mutant strain. Real time qPCR data demonstrated that under salt and sorbitol stress conditions the Fphog1 gene is not subject of transcriptional regulation. Levels of reactive oxygen species (ROS), mitochondrial membrane permeability transition, nuclear disintegration and DNA fragmentation, indicators of programmed cell death (PCD) all showed significant increases under osmotic stress conditions in the Delta Fphog1 mutant in comparison to the wild type strain. These results suggest that an important function of Fphog1 is attenuating apoptotic phenotypes under salt and sorbitol stressors. PMID- 18506896 TI - Production of an extracellular thermohalophilic lipase from a moderately halophilic bacterium, Salinivibrio sp. strain SA-2. AB - Fifty strains of moderately halophilic bacteria were isolated from various salty environments in Iran. A strain designated as SA-2 was shown to be the best producer of extracellular lipase and was selected for further studies. Biochemical and physiological characterization along with 16S rDNA sequence analysis placed SA-2 in the genus Salinivibrio. The optimum salt, pH, temperature and aeration for enzyme production were 0.1 M KCl, pH 8, 35 degrees C and 150 rpm, respectively. The enzyme production was synchronized bacterial growth and reached a maximum level during the early-stationary phase in the basal medium containing 1 M NaCl. Triacylglycerols enhanced lipase production, while carbohydrates had inhibitory effects on it. The maximum lipase activity was obtained at pH 7.5, 50 degrees C and CaCl(2) concentration of 0.01 M. The enzyme was stable at pH range of 7.5-8 and retained 90% of its activity at 80 degrees C for 30 min. Different concentrations of NaNO(3), Na(2)SO(4), KCl and NaCl had no affect on lipase stability for 3 h. These results suggest that the lipase secreted by Salinivibrio sp. strain SA-2 is industrially important from the perspective of its tolerance to a broad temperature range, its moderate thermoactivity and its high tolerance to a wide range of salt concentrations (0-3 M NaCl). PMID- 18506897 TI - Coordinate regulation of gallbladder motor function in the gut-liver axis. AB - Gallstones are one of the most common digestive diseases with an estimated prevalence of 10%-15% in adults living in the western world, where cholesterol enriched gallstones represent 75%-80% of all gallstones. In cholesterol gallstone disease, the gallbladder becomes the target organ of a complex metabolic disease. Indeed, a fine coordinated hepatobiliary and gastrointestinal function, including gallbladder motility in the fasting and postprandial state, is of crucial importance to prevent crystallization and precipitation of excess cholesterol in gallbladder bile. Also, gallbladder itself plays a physiopathological role in biliary lipid absorption. Here, we present a comprehensive view on the regulation of gallbladder motor function by focusing on recent discoveries in animal and human studies, and we discuss the role of the gallbladder in the pathogenesis of gallstone formation. PMID- 18506898 TI - Increased risk of hepatocellular carcinoma among patients with hepatitis C cirrhosis and diabetes mellitus. AB - Recent studies suggest that diabetes mellitus increases the risk of developing hepatocellular carcinoma (HCC). The aim of this study is to quantify the risk of HCC among patients with both diabetes mellitus and hepatitis C in a large cohort of patients with chronic hepatitis C and advanced fibrosis. We included 541 patients of whom 85 (16%) had diabetes mellitus. The median age at inclusion was 50 years. The prevalence of diabetes mellitus was 10.5% for patients with Ishak fibrosis score 4, 12.5% for Ishak score 5, and 19.1% for Ishak score 6. Multiple logistic regression analysis showed an increased risk of diabetes mellitus for patients with an elevated body mass index (BMI) (odds ratio [OR], 1.05; 95% confidence interval [CI], 1.00-1.11; P = 0.060) and a decreased risk of diabetes mellitus for patients with higher serum albumin levels (OR, 0.81; 95% CI, 0.63 1.04; P = 0.095). During a median follow-up of 4.0 years (interquartile range, 2.0-6.7), 11 patients (13%) with diabetes mellitus versus 27 patients (5.9%) without diabetes mellitus developed HCC, the 5-year occurrence of HCC being 11.4% (95% CI, 3.0-19.8) and 5.0% (95% CI, 2.2-7.8), respectively (P = 0.013). Multivariate Cox regression analysis of patients with Ishak 6 cirrhosis showed that diabetes mellitus was independently associated with the development of HCC (hazard ratio, 3.28; 95% CI, 1.35-7.97; P = 0.009). CONCLUSION: For patients with chronic hepatitis C and advanced cirrhosis, diabetes mellitus increases the risk of developing HCC. PMID- 18506899 TI - Isolation, characterization of heavy metal resistant strain of Pseudomonas aeruginosa isolated from polluted sites in Assiut city, Egypt. AB - Sixty six isolates of Pseudomonas spp. were isolated from wastewater of El-Malah canal located in Assiut, Egypt and were checked for their heavy metal tolerance. One isolate has tested for its multiple metal resistances and found to be plasmid mediated with molecular weight 27 Kb for nickel and lead. It was identified as Pseudomonas aeruginosa ASU 6a. Its minimal inhibitory concentration (MIC) for Cu(2+), Co(2+), Ni(2+), Zn(2+), Cr(3+), Cd(2+)and Pb(2+) were 6.3, 5.9, 6.8, 9.2, 5.8, 4.4, and 3.1 mM, respectively. Growth kinetics and the maximum adsorption capacities were determined under Ni(2+) and Pb(2+) stress. The latter heavy metals induced potassium efflux and were used as indicator for plasma membrane permeabilization. PMID- 18506900 TI - Nucleotide sequence of plasmid pA387 of Amycolatopsis benzoatilytica and construction of a conjugative shuttle vector. AB - The complete nucleotide sequence of plasmid pA387 of Amycolatopsis benzoatilytica DSM 43387 was determined. Sequence analysis revealed that pA387 is 30,157 bp long and has a G+C content of 71.74%. To obtain a minimal transferable replicon capable of self-replication, a 2,176 bp fragment of pA387 was cloned, and we demonstrated that this fragment is sufficient for autonomous replication. The replication region of pA387 exhibited no significant homology to any known replication proteins available in databases. Putative maintenance and transfer functions were identified on pA387. The predicted products of open reading frames, ORF 2 and ORF 12, resembled the plasmid stabilizing proteins, a DNA resolvase and a ParA protein, respectively. The putative translational products of ORF 15 and ORF 16 showed similarity to known bacterial conjugation proteins, TraG and TraA, respectively. A conjugative Escherichia coli -Amycolatopsis shuttle-cloning vector was constructed by using the pA387 replicon and designated pSETRL1. Shuttle vector pSETRL1 successfully transformed Amycolatopsis mediterranei DSM 40773 and Amycolatopsis orientalis NBRC 12806 by conjugation and electroporation, and is likely to be a useful vector in Amycolatopsis research. PMID- 18506901 TI - Direct TLC resolution of atenolol and propranolol into their enantiomers using three different chiral selectors as impregnating reagents. AB - Direct resolution of racemic atenolol and propranolol into their enantiomers was achieved by normal phase TLC on silica gel plates impregnated with optically pure L-tartaric acid, (R)-mandelic acid and (-)-erythromycin as chiral selectors. Different solvent systems were worked out to resolve the enantiomers. Spots were detected using iodine vapour. The TLC method was validated for linearity, limit of detection and limit of quantification. The influence of pH, temperature and concentration of chiral selector was studied. PMID- 18506902 TI - Analysis of Cibacron blue F3G-A interaction with therapeutic proteins by MALDI TOF mass spectrometry. AB - The formation of the complexes between Cibacron blue F3G-A and two therapeutic proteins, recombinant human interferon-alpha2b and recombinant human growth hormone, was investigated. The method of time-resolved limited proteolysis coupled with MALDI-TOF mass spectrometry was used. The analysis of peptide maps revealed that A(17)HR(19) and L(20)HQLAFDTYQEFEEAYIPK(38) of hGH, and R(14)TLMLLAQMR(23) and D(33)RHDFGFPQEEFGNQFQK(50) of hIFN-alpha2b, exhibit affinity to Cibacron blue F3G-A. PMID- 18506903 TI - Evaluation of fungicidal activity of extracellular filtrates of cyanobacteria- possible role of hydrolytic enzymes. AB - A set of seventy axenised and unicyanobacterial isolates belonging to the genus Anabaena were evaluated for biocidal activity against a set of phytopathogenic fungi. Among them, 35 Anabaena strains showed zone of inhibition against one or more fungi. The extracellular filtrates from 4 and 8 weeks old cultures of these Anabaena strains were further evaluated in terms of hydrolytic enzymes, proteins and IAA employing standard methods. Significant differences were also observed among the strains in terms of their FPase, chitosanase and xylanase activity, while low and relatively similar values of CMCase, cellobiase and protease activity were recorded in the strains analyzed. IAA production was also observed in all the strains. Comparative evaluation of activity of hydrolytic enzymes and antifungal activity revealed that such enzymes may contribute to the fungicidal activity of the cyanobacterial strains, besides other bioactive compounds, including IAA, which are established promising traits for biocontrol agents. This study is a first time report on the production of hydrolytic enzymes by these oxygenic photosynthetic prokaryotes, which can be potential candidates for the development of biocontrol agent(s) against selected phytopathogenic fungi. PMID- 18506904 TI - Biosorption of copper by yeast, Loddermyces elongisporus, isolated from industrial effluents: its potential use in wastewater treatment. AB - The present study is aimed at assessing the ability of metal resistant yeast, Loddermyces elongisporus, to uptake metal from liquid medium. The minimum inhibitory concentration of Cu(2+) against Loddermyces elongisporus ranged between 2.2-2.3 mg/l. The yeast could also tolerate Zn(2+) (2.9 mg/l), Hg(2+) (2.4 mg/l), Ni(2+) (2.2 mg/l), Cr(6+) (2.0 mg/l), Pb(2+) (1.1 mg/l), and Cd(2+) (0.8 mg/l). The yeast isolate showed typical growth curves but lag and log phases extended in the presence of copper. Yeast isolate showed optimum growth at 30 degrees C and pH 8. Metal processing ability of the isolate was determined in a medium containing 0.1 mg/l of Cu(2+). Loddermyces elongisporus could reduce Cu(2+) 15%, 26%, 39%, 50%, 60%, 67%, 75% and 81% from the medium after 6, 12, 18, 24, 30, 48, 72 and 96 hours, respectively. L. elongisporus could also efficiently remove 80% copper from the medium after 96 h and was able to remove Cu(2+) 60% and 77% from the wastewater after 4 and 8 d, respectively. The metal binding ability suggests possibility of using this yeast strain for removal of copper from metal contaminated wastewater. PMID- 18506905 TI - Peptide inhibitors of MurD and MurE, essential enzymes of bacterial cell wall biosynthesis. AB - Continuous development of antibacterial compounds with novel modes of action (accompanied by rationalization of chemotherapeutic prescription) is the best way to address the growing problem of antibiotic-resistant infections. Numerous clinically important antibiotics interfere with peptidoglycan cell wall biosynthesis making this unique metabolic pathway a well validated target for antimicrobials. While nearly all of these antibiotics inhibit late stages of murein synthesis occurring on the extracellular side of plasma membrane, initial cytoplasmic steps have not been extensively exploited as drug targets. We performed affinity selection of peptides from phage-displayed libraries against two essential bacterial enzymes MurD and MurE involved in the cytoplasmic synthesis of peptidoglycan monomer. Selected peptides were found to inhibit respective target enzymes in an in vitro assay with IC(50) values of 140 microM to 1.5 mM. These peptides represent starting point for design of peptidomimetic lead compounds with the ultimate objective of small molecule chemotherapeutic development. PMID- 18506906 TI - Isolation and identification of a psychrotrophic Acinetobacter sp. CR9 and characterization of its alkaline lipase. AB - Forty three psychrotrophic bacteria were isolated from soil samples collected from Chandra river in sub-alpine region of western Himalaya, India. Among these, 11 isolates were found positive for lipase production at low temperature. Of 11 isolates, CR9 produced largest zone of clearance on plate assay and was able to produce lipase under wide range of pH. The isolate CR9 was identified as Acinetobacter sp. based on morphological and physiochemical characterization and 16S rRNA gene sequencing analysis. According to 16S rRNA gene sequencing data the closest phylogenetic neighbor for strain CR9 was Acinetobacter lwoffii (98.9%). The partially purified lipase from strain CR9 exhibited maximum activity at temperature 40 degrees C and pH optima at 8.0. Cu(2+), Mo(2+), Mg(2+), Zn(2+), phenylmethanesulfonyl fluoride (PMSF), dithiothreitol (DTT) and beta mercaptoethanol (2-ME) enhanced the enzyme activity, whereas Ca(2+) and ethylenediaminetetraacetic acid (EDTA) had inhibitory effect. Lipase hydrolyzed wide range of short chain fatty acid esters of p-nitrophenyl. The organism CR9 also hydrolyzed tributyrin, Tween 80, soybean oil, mustard oil and olive oil. The results highlight the relevance of unexplored microbes from cold environments of western Himalaya for the isolation of novel lipase producing bacteria. PMID- 18506907 TI - Susceptibility of local Agrobacterium tumefaciens strains to streptomycetes isolates from Jordan soils. AB - Five strains of Agrobacterium tumefaciens were isolated and characterized from 12 different plant tumors. The susceptibility of these phytopathogens to antibiotics and to soil Streptomyces isolates was tested. Among the 90 Streptomyces isolates, only 12 were able to inhibit the growth of at least one A. tumefaciens strain. Four strains of A. tumefaciens were susceptible to streptomycin and cefotaxime. In addition, Streptomyces 404 strain was able to inhibit the growth of four strains of the Agrobacterium pathogens with an inhibition zone diameter ranged between 10 and 16 mm. The strong inhibitory effects of Streptomyces 404 strain on A. tumefaciens suggest the use of this strain as a promising agent to control crown gall disease. PMID- 18506908 TI - Identification and antioxidant activity of melanin isolated from Hypoxylon archeri, a companion fungus of Tremella fuciformis. AB - Melanin produced by Hypoxylon archeri, a companion fungus of Tremella fuciformis, was purified from the submerged culture medium and designated as HM. Ultraviolet visible and FTIR spectra of the purified HM showed significant similarities with those of the synthesized melanin derived from tyrosine. Thus, the HM melanin was identified to be derived from the precursor molecule of tyrosine or dihydroxyphenylalanine (DOPA). The antioxidant activity of HM melanin, synthetic melanin and vitamin C were compared by inhibition of the oxidation of 5-thio-2 nitrobenzoic acid (TNB) caused by hypochlorous acid (HOCl) and hydrogen peroxide (H(2)O(2)). The HM melanin is likely to be more efficient eliminating oxygen free radicals generated by H(2)O(2) than by HOCl. At a concentration of 100 mg x l( 1), the HM melanin protected 80.95% of TNB oxidation by H(2)O(2), slightly higher than the synthetic melanin. Inhibition curves as a function of time also revealed the HM melanin was a more efficient inhibitor of H(2)O(2) oxidation with an average TNB (TNB(acr)) consumption rate of 0.0553 mmol l(-1 ) x min(-1 )during the inhibition phase. Therefore, HM melanin is an efficient scavenger of peroxide free radicals. PMID- 18506909 TI - Enhanced electrochemiluminescence of Ru(bpy)3(2+) by flavone compounds. AB - Flavones such as morin, rutin, quercitrin, quercetin and wogonin were found to be able to strongly enhance the electrochemiluminescence (ECL) of the Ru(bpy)(3)(2+) system. Based on this, a novel ECL method with good stability and reproducibility could be developed for determination of flavones. Under the optimum conditions, the enhanced ECL intensity was linear with the flavones concentration in a wide range. The detection limits (defined as S:N = 3) for morin, rutin, quercitrin, quercetin and wogonin were 3.2 x 10(-7) mol/L, 4.3 x 10(-7), 1.8 x 10(-7), 8.0 x 10(-8) and 1.0 x 10(-7 ) mol/L, respectively. In addition, the possible mechanism for the Ru(bpy)(3)(2+) ECL system in the presence of flavones is also discussed. PMID- 18506910 TI - An HPLC method for the determination and pharmacokinetic study of lehmannine in rat plasma. AB - A high-performance liquid chromatographic (HPLC) method for determining lehmannine (LMN) in rat plasma was developed for application in the pharmacokinetics study. The plasma was deproteinized with acetonitrile that contained an internal standard and was separated from the aqueous layer by adding sodium chloride. The HPLC assay was carried out using a VP-ODS column at 40 degrees C. The mobile phase was acetonitrile-0.02 mol/L ammonium acetate buffer triethylamine (35:65:0.04, v/v/v). The flow rate was 1.0 mL/min. The detection wavelength was set at 220 nm. The method was used to determine the concentration time profiles of LMN in the plasma following oral administration or bolus injection of LMN aqueous solution. The pharmacokinetic parameters of LMN were calculated for the first time by Drug and Statistics 1.0 program. PMID- 18506911 TI - A sensitive method for determination of salvianolic acid A in rat plasma using liquid chromatography/tandem mass spectrometry. AB - Salvianolic acid A (SAA), a major effective constituent of Salvia miltiorrhizas, is widely used in traditional Chinese medicine. A sensitive rapid analytical method was established and validated for SAA in rat plasma, which was further applied to assess the pharmacokinetics of SAA in rats receiving a single oral dose of SAA. The method used liquid chromatography tandem mass spectrometry in multiple reaction monitoring mode with chloramphenicol as the internal standard. A simple liquid-liquid extraction based on ethyl acetate was employed. The combination of a simple sample cleanup and short chromatographic run time (3 min) increased the throughput of the method substantially. The method was validated over the range 1.4-1000 ng/mL with a correlation coefficient >0.99. The lower limit of quantification was 1.4 ng/mL for SAA in plasma. Intra- and inter-day accuracies for SAA were 95-113 and 98-107%, and the inter-day precision was less than 12%. This method is more sensitive and faster than previous methods. After a single oral dose of 100 mg/kg of SAA, the mean peak plasma concentration (Cmax) of SAA was 318 ng/mL at 0.5 h, the area under the plasma concentration-time curve (AUC0-12 h) was 698 +/- 129 ng.h/mL, and the elimination half-life (T1/2) was 3.29 h. PMID- 18506912 TI - Changes in IL-8 release and intracellular content in DMSO-differentiated HL-60 cells after treatment with 4-hydroxynonenal. AB - 4-Hydroxynonenal (HNE), a chemotactic aldehyde produced by lipid peroxidation, has been shown to trigger exocytosis in HL-60 cells induced to differentiate toward the granulocytic cell line by DMSO. In this work we studied HNE effects on the intracellular content of IL-8 and its release in DMSO-differentiated HL-60 cells. Cell incubation at 37 degrees C in the presence of 0.1 microM HNE induced a significant increase of IL-8 release after 30 min; the degree of HNE-induced IL 8 secretion became quite strong after 1 h, whereas the intracellular content showed no statistically significant changes. By contrast, 1 microM HNE induced a low decrease of the chemokine release; however, the used HNE concentrations failed to increase the release of lactate dehydrogenase (LDH), a test used to assay cell viability. The addition of 0.1 microM IL-8 to DMSO-differentiated HL 60 cells induced a strong increase of exocytosis, measured by beta-glucuronidase secretion. Exocytosis stimulation by IL-8 was much higher than that given by the aldehyde; the addition of various HNE concentrations to cells incubated in the presence of IL-8 decreased the secretion given by the cytokine alone. However, HNE-induced exocytosis was likely to be a direct action of the aldehyde and was not mediated through the stimulation of IL-8 release since HNE was unable to modify IL-8 secretion during the short time of 10 min used in the exocytosis assay. PMID- 18506913 TI - Galantamine-induced pisa syndrome: memantine as an alternative. PMID- 18506914 TI - Liver transplantation: yesterday, today and tomorrow. AB - With the advances in technical skills, management of postoperative complications and improvements in immunosuppressive drugs, liver transplantation is the standard treatment for many patients with chronic liver disease. Today, shortage of donor organs seems to be the major limiting factor for the application of liver transplantation. This review focuses on five issues that are challenging to clinical practice of liver transplantation and relevant to gastroenterologists. These include living donor liver transplantation, recurrent viral hepatitis, non heart-beating donors, hepatocellular carcinoma, and ABO incompatible liver transplantation. Living donor and non-heart beating donor transplantations were initiated as a solution to increase the donor organ pool and it is expected that there will be an increase in the number of these donors. Recurrent hepatitis C and hepatocellular carcinoma following liver transplantation are among major problems and ongoing research in these diseases may lead to better outcomes in these recipients. PMID- 18506915 TI - Strategy for treatment of nonerosive reflux disease in Asia. AB - The paper is to review the clinical and pathophysiologic differences between of nonerosive reflux disease (NERD) and reflux esophagitis (RE), and to propose a treatment strategy for NERD, especially for patients in Asia. A Medline search was performed regarding the clinical and pathophysiologic differences between NERD and RE, and treatment of NERD and RE. The characteristics of NERD patients in Asia are as follows: (1) high proportion of female patients, (2) low frequency of hiatal hernia, (3) high frequency of H pylori infection, (4) severe glandular atrophy of the gastric mucosa, and (5) frequent resistance to proton pump inhibitor (PPI) therapy. In Asian NERD patients, exposure of the esophagus to acid is not increased, and esophageal motility is normal. These characteristics are similar to those of patients in Western countries. Our recommended first choice treatment is administration of PPI in combination with a prokinetic agent. However, at present, because there is limited evidence regarding effective treatments for NERD, it is best to try several different treatment strategies to find the best treatment for each patient. PMID- 18506916 TI - Intraductal biliary and pancreatic endoscopy: an expanding scope of possibility. AB - Intraductal endoscopy describes the use of an endoscope to directly visualize the biliary and pancreatic ducts. For many years, technological challenges have made performing these procedures difficult. The "mother-baby" system and other various miniscopes have been developed, but routine use has been hampered due to complex setup, scope fragility and the time consuming, technically demanding nature of the procedure. Recently, the SpyGlass peroral cholangiopancreatoscopy system has shown early success at providing diagnostic information and therapeutic options. The clinical utility of intraductal endoscopy is broad. It allows better differentiation between benign and malignant processes by allowing direct visualization and targeted sampling of tissue. Therapeutic interventions, such as electrohydraulic lithotripsy (EHL), laser lithotripsy, photodynamic therapy, and argon plasma coagulation (APC), may also be performed as part of intraductal endoscopy. Intraductal endoscopy significantly increases the diagnostic and therapeutic yield of standard endoscopic retrograde cholangiography (ERCP), and as technology progresses, it is likely that its utilization will only increase. In this review of intraductal endoscopy, we describe in detail the various endoscopic platforms and their diagnostic and clinical applications. PMID- 18506917 TI - Pain management in chronic pancreatitis. AB - Abdominal pain is a major clinical problem in patients with chronic pancreatitis. The cause of pain is usually multifactorial with a complex interplay of factors contributing to a varying degree to the pain in an individual patient and, therefore, a rigid standardized approach for pain control tends to lead to suboptimal results. Pain management usually proceeds in a stepwise approach beginning with general lifestyle recommendations. Low fat diet, alcohol and smoking cessation are encouraged. Analgesics alone are needed in almost all patients. Maneuvers aimed at suppression of pancreatic secretion are routinely tried. Patients with ongoing symptoms may be candidates for more invasive options such as endoscopic therapy, and resective or drainage surgery. The role of pain modifying agents (antidepressants, gabapentin, peregabalin), celiac plexus block, antioxidants, octreotide and total pancreatectomy with islet cell auto transplantation remains to be determined. PMID- 18506918 TI - Pancreatic function testing: here to stay for the 21st century. AB - The diagnosis of Chronic Pancreatitis (CP) is based on the detection of abnormal structure or function of the diseased pancreas. The pancreatic function tests more accurately determine the presence of CP than tests of structure, especially for early stage disease. The function tests can be divided into two categories: non-invasive and invasive. The invasive "tube" tests can reliably detect mild, early CP, but are only available at a few referral centers and tend to be poorly tolerated by patients. The non-invasive tests are easy to obtain, but tend to perform poorly in patients with early, mild disease. Therefore, no one test is useful in all clinical situations, and a detailed understanding of the rational, pathophysiologic basis, strengths, and limitations of various tests is needed. This review highlights the role of various pancreatic function tests in the diagnosis of CP including fecal fat analysis, fecal elastase, fecal chymotrypsin, serum trypsin, the secretin stimulation test, the cholecystokinin (CCK) stimulation test, the combined secretin-CCK stimulation test, the intraductal and endoscopic secretin stimulation tests, and the functional magnetic resonance imaging of the pancreas after secretin stimulation. PMID- 18506919 TI - Analysis of surgical and perioperative complications in seventy-five right hepatectomies for living donor liver transplantation. AB - AIM: To present an analysis of the surgical and perioperative complications in a series of seventy-five right hepatectomies for living-donation (RHLD) performed in our center. METHODS: From January 2002 to September 2007, we performed 75 RHLD, defined as removal of a portion of the liver corresponding to Couinaud segments 5-8, in order to obtain a graft for adult to adult living-related liver transplantation (ALRLT). Surgical complications were stratified according to the most recent version of the Clavien classification of postoperative surgical complications. The perioperative period was defined as within 90 d of surgery. RESULTS: No living donor mortality was present in this series, no donor operation was aborted and no donors received any blood transfusion. Twenty-three (30.6%) living donors presented one or more episodes of complication in the perioperative period. Seven patients (9.33%) out of 75 developed biliary complications, which were the most common complications in our series. CONCLUSION: The need to define, categorize and record complications when healthy individuals, such as living donors, undergo a major surgical procedure, such as a right hepatectomy, reflects the need for prompt and detailed reports of complications arising in this particular category of patient. Perioperative complications and post resection liver regeneration are not influenced by anatomic variations or patient demographic. PMID- 18506920 TI - Treatment and survival in a population-based sample of patients diagnosed with gastroesophageal adenocarcinoma. AB - AIM: To examine the extent of use of specific therapies in clinical practice, and their relationship to therapies validated in clinical trials. METHODS: The US National Cancer Institutes' Patterns of Care study was used to examine therapies and survival of patients diagnosed in 2001 with histologically-confirmed gastroesophageal adenocarcinoma (n = 1356). The study re-abstracted data and verified therapy with treating physicians for a population-based stratified random sample. RESULTS: Approximately 62% of patients had stomach adenocarcinoma (SAC), while 22% had gastric-cardia adenocarcinoma (GCA), and 16% lower esophageal adenocarcinoma (EAC). Stage IV/unstaged esophageal cancer patients were most likely and stage I-III stomach cancer patients least likely to receive chemotherapy as all or part of their therapy; gastric-cardia patients received chemotherapy at a rate between these two. In multivariable analysis by anatomic site, patients 70 years and older were significantly less likely than younger patients to receive chemotherapy alone or chemoradiation for all three anatomic sites. Among esophageal and stomach cancer patients, receipt of chemotherapy was associated with lower mortality; but no association was found among gastric cardia patients. CONCLUSION: This study highlights the relatively low use of clinical trials-validated anti-cancer therapies in community practice. Use of chemotherapy-based treatment was associated with lower mortality, dependent on anatomic site. Findings suggest that physicians treat lower esophageal and SAC as two distinct entities, while gastric-cardia patients receive a mix of the treatment strategies employed for the two other sites. PMID- 18506921 TI - In vitro activity of moxifloxacin and piperacillin/sulbactam against pathogens of acute cholangitis. AB - AIM: To analyze the in vitro activity of moxifloxacin and piperacillin/sulbactam against pathogens isolated from patients with acute cholangitis. METHODS: In this prospective study a total of 65 patients with acute cholangitis due to biliary stone obstruction (n = 7), benign biliary stricture (n = 16), and malignant biliary stricture (n = 42) were investigated with regard to spectrum of bacterial infection and antibiotic resistance. Pathogens were isolated from bile cultures in all study patients. In 22 febrile patients, blood cultures were also obtained. In vitro activity of moxifloxacin and piperacillin/sulbactam was determined by agar diffusion. RESULTS: Thirty-one out of 65 patients had positive bile and/or blood cultures. In 31 patients, 63 isolates with 17 different species were identified. The predominant strains were Enterococcus species (26/63), E.coli (13/63) and Klebsiella species (8/63). A comparable in vitro activity of moxifloxacin and piperacillin/sulbactam was observed for E.coli and Klebsiella species. In contrast, Enterococcus species had higher resistances towards moxifloxacin. Overall bacteria showed antibiotic resistances in vitro of 34.9% for piperacillin/sulbactam and 36.5% for moxifloxacin. CONCLUSION: Enterococcus species, E.coli and Klebsiella species were the most common bacteria isolated from bile and/or blood from patients with acute cholangitis. Overall, a mixed infection with several species was observed, and bacteria showed a comparable in vitro activity for piperacillin/sulbactam and moxifloxacin. PMID- 18506922 TI - Are acute exacerbations of chronic inflammatory appendicitis triggered by coprostasis and/or coproliths? AB - AIM: To examine the role of coprostasis and coproliths in recurrent appendicitis. METHODS: We evaluated four hundred and twenty seven consecutive pathology reports of all appendectomy specimens from January 2003 to December 2004. Findings were categorised as showing acute appendicitis, acute recurrent appendicitis, subacute recurrent appendicitis, chronic appendicitis, or appendices without inflammation. All patients had presented with acute right lower quadrant pain. In 94 instances, there was a history of recurrent similar episodes in the past. RESULTS: Of the 427 histology reports, 294 were inter-preted as showing acute appendicitis, 56 acute recurrent appendicitis, 34 subacute recurrent appen-dicitis, 28 chronic appendicitis, and 15 non-inflamed appendices. Coprostasis was observed in 58 patients (13.58%) and the presence of coprolith in 6 (1.4%). Coprostasis, and age, were among the predictors in the final model. CONCLUSION: Coprostasis but not coproliths seems to be a contributing factor to acute exacerbations of chronic inflammatory appendicitis. PMID- 18506923 TI - Retrospective analysis of old-age colitis in the Dutch inflammatory bowel disease population. AB - AIM: To describe the characteristics of Dutch patients with chronic inflammatory bowel disease (IBD) first diagnosed above 60 years of age-a disease also known as old-age colitis (OAC) and to highlight a condition that has a similar appearance to IBD, namely segmental colitis associated with diverticular disease (SCAD). METHODS: A retrospective longitudinal survey of patient demographic and clinical characteristics, disease characteristics, diagnostic methods, management and course of disease was performed. The median follow-up period was 10 years. RESULTS: Of a total of 1100 IBD patients attending the Department of Gastroenterology, 59 (5%) [median age 82 years (range 64-101); 25 male (42%)] were identified. These patients were diagnosed with ulcerative colitis (n = 37, 61%), Crohn's disease (n = 14, 24%), and indeterminate colitis (n = 8, 15%). Remission was induced in 40 (68%) patients within a median interval of 6 mo (range 1-21) and immunosuppressive therapy was well tolerated. Histological evaluation based on many biopsy samples and the course of the disease led to other diagnosis, namely SCAD instead of IBD in five (8%) patients. CONCLUSION: OAC is not an infrequent problem for the gastroenterologist, and should be considered in the evaluation of older patients with clinical features suggestive of IBD. Extra awareness and extensive biopsy sampling are required in order to avoid an erroneous diagnosis purely based on histological mimicry of changes seen in SCAD, when diagnosing IBD in the presence of diverticulosis coli. PMID- 18506924 TI - Effect of probiotic Lactobacillus rhamnosus GG intervention on global serum lipidomic profiles in healthy adults. AB - AIM: To investigate the effect of three weeks' intervention with a probiotic Lactobacillus rhamnosus GG (LGG) bacteria on global serum lipidomic profiles and evaluate whether the changes in inflammatory variables (CRP, TNF-alpha and IL-6) are reflected in the global lipidomic profiles of healthy adults. METHODS: We performed UPLC/MS-based global lipidomic platform analysis of serum samples (n = 26) in a substudy of a randomised, double-blind, placebo-controlled 3-wk clinical intervention trial investigating the immunomodulatory effects of probiotics in healthy adults. RESULTS: A total of 407 lipids were identified, corresponding to 13 different lipid classes. Serum samples showed decreases in the levels of lysophosphatidylcholines (LysoGPCho), sphingomyelins (SM) and several glycerophosphatidylcholines (GPCho), while triacylglycerols (TAG) were mainly increased in the probiotic LGG group during the intervention. Among the inflammatory variables, IL-6 was moderately associated by changes in global lipidomic profiles, with the top-ranked lipid associated with IL-6 being the proinflammatory LysoGPCho (20:4). There was a weak association between the lipidomic profiles and the two other inflammatory markers, TNF-alpha and CRP. CONCLUSION: This was the first study to investigate the effects of probiotic intervention on global lipidomic profiles in humans. There are indications that probiotic LGG intervention may lead to changes in serum global lipid profiles, as reflected in decreased GPCho, LysoGPCho and SM as well as mainly increased TAG. PMID- 18506925 TI - Factors that influence outcome in non-invasive and invasive treatment in polycystic liver disease patients. AB - AIM: To evaluate the factors that influence outcome of both non-invasive and invasive treatment of polycystic liver disease. METHODS: Analysis of clinical files of patients with complete follow-up from July 1986 to June 2006. RESULTS: Forty-one patients (male, 7; female, 34), 47.8 +/- 11.9 years age, and 5.7 +/- 6.7 years follow-up, were studied. Alkaline phosphatase (AP) elevation (15% of patients) was associated with the requirement of invasive treatment (IT, P = 0.005). IT rate was higher in symptomatic than non-symptomatic patients (65.4% vs 14.3%, P = 0.002), and in women taking hormonal replacement therapy (HRT) (P = 0.001). Cysts complications (CC) were more frequent (22%) in the symptomatic patients group (P = 0.023). Patients with body mass index (BMI) > 25 (59%) had a trend to complications after IT (P = 0.075). Abdominal pain was the most common symptom (56%) and indication for IT (78%). Nineteen patients (46%) required a first IT: 12 open fenestration (OF), 4 laparoscopic fenestration (LF) and 3 fenestration with hepatic resection (FHR). Three required a second IT, and one required a third procedure. Complications due to first IT were found in 32% (OF 16.7%, LF 25%, FHR 66.7%), and in the second IT in 66.7% (OF 100%). Follow-up mortality rate was 0. CONCLUSION: Presence of symptoms, elevated AP, and CC are associated with IT requirement. HRT is associated with presence of symptoms and IT requirement. Patients with BMI > 25 have a trend be susceptible to IT complications. The proportions of complications are higher in FHR and second IT groups. RS is more frequent after OF. PMID- 18506926 TI - Hepatitis B virus prevalence and transmission risk factors in inflammatory bowel disease patients at Clementino Fraga Filho university hospital. AB - AIM: To evaluate the prevalence of hepatitis B virus (HBV) infection in inflammatory bowel disease (IBD) patients that followed up in our hospital and try to identify the possible risk factors involved in this infection transmission. METHODS: This was a cross-sectional study for which 176 patients were selected according to their arrival for the medical interview. All these patients had already IBD diagnosis. The patient was interviewed and a questionnaire was filled out. RESULTS: In the group of 176 patients whom we examined, we found that 17% (30) were anti-HBc positive. Out of 30 patients with positive anti-HBc, 2.3% (4) had positive HBsAg and negative HBV-DNA. In an attempt to identify the possible HBV infection transmission risk factors in IBD patients, it was observed that 117 patients had been submitted to some kind of surgical procedure, but only 24 patients had positive anti-HBc (P = 0.085). It was also observed that surgery to treat IBD complications was not a risk factor for HBV infection transmission, since we did not get a statically significant P value. However, IBD patients that have been submitted to surgery to treat IBD complications received more blood transfusions then patients submitted to other surgical interventions (P = 0.015). CONCLUSION: There was a high incidence of positive anti-HBc (17%) and positive HBsAg (2.3%) in IBD patient when compared with the overall population (7.9%). PMID- 18506927 TI - Contrast-enhanced intraoperative ultrasonography equipped with late Kupffer-phase image obtained by sonazoid in patients with colorectal liver metastases. AB - AIM: To find occult metastases during hepatectomy in patients with colorectal cancer liver metastases (CRCLM), contrast-enhanced intraoperative ultrasonography (CE-IOUS) was performed using a new microbubble agent, sonazoid, which provides a parenchyma-specific contrast image based on its accumulation in the Kupffer cells. METHODS: Eight patients with CRCLM underwent CE-IOUS using sonazoid before hepatectomy. The liver was investigated during a late Kupffer-phase imaging, which is a valuable characteristic of sonazoid. RESULTS: CE-IOUS using sonazoid provided the early vascular- and sinusoidal-phase images for 10 min followed by the late Kupffer-phase image up to 30 min after the injection of sonazoid. IOUS did not provide new findings of metastatic lesion in the 8 patients. However, during the late Kupffer-phase image of sonazoid, a metastatic lesion was newly found in two of the 8 patients. These newly detected lesions were removed by an additional hepatectomy and histopathologically diagnosed as a metastasis. CONCLUSION: CE-IOUS using sonazoid can allow surgeons to investigate the whole liver with enough time and to find new metastases intraoperatively. PMID- 18506928 TI - Incidence of reflux esophagitis and H pylori infection in diabetic patients. AB - AIM: To investigate the incidence of reflux esophagitis (RE) and H pylori infection in the diabetic patient. METHODS: The incidence of RE and H pylori infection were investigated in 85 patients with diabetes mellitus and the results were compared with controls. RESULTS: The incidence of RE in diabetic patients was 17.6%. Although this tended to be higher in diabetic patients, there were no statistically significant differences between diabetic patients and controls. The incidence of H pylori infection in diabetic patients was 53.7% but no statistically significant difference was seen between diabetic patients and controls in the incidence of H pylori infection. CONCLUSION: No significant differences could be seen between diabetic patients and controls in the incidence of RE and H pylori infection. PMID- 18506929 TI - Reactive oxygen species and chemokines: are they elevated in the esophageal mucosa of children with gastroesophageal reflux disease? AB - AIM: To determine the role of inflammatory cytokines and reactive oxygen species (ROS) in childhood reflux esophagitis. METHODS: A total of 59 subjects who had complaints suggesting GERD underwent esophagogastroduodenoscopy. Endoscopic and histopathologic diagnosis of reflux esophagitis was established by Savary-Miller and Vandenplas grading systems, respectively. Esophageal biopsy specimens were taken from the esophagus 20% proximal above the esophagogastric junction for conventional histopathological examination and the measurements of ROS and cytokine levels. ROS were measured by chemiluminescence, whereas IL-8 and MCP-1 levels were determined with quantitative immunometric ELISA on esophageal tissue. Esophageal tissue ROS, IL-8 and MCP-1 levels were compared among groups with and without endoscopic/histo-pathologic esophagitis. RESULTS: Of 59 patients 28 (47.5%) had normal esophagus whereas 31 (52.5%) had endoscopic esophagitis. In histopathological evaluation, almost 73% of the cases had mild and 6.8% had moderate degree of esophagitis. When ROS and chemokine levels were compared among groups with and without endoscopic esophagitis, statistical difference could not be found between patients with and without esophagitis. Although the levels of ROS, IL-8 and MCP-1 were found to be higher in the group with histopathological reflux esophagitis, this difference was not statistically significant. CONCLUSION: These results suggest that the grade of esophagitis is usually mild or moderate during childhood and factors apart from ROS, IL-8 and MCP-1 may be involved in the pathogenesis of reflux esophagitis in children. PMID- 18506930 TI - Prognostic factors in patients with advanced cholangiocarcinoma: role of surgery, chemotherapy and body mass index. AB - AIM: To study the factors that may affect survival of cholangiocarcinoma in Lebanon. METHODS: A retrospective review of the medical records of 55 patients diagnosed with cholangio-carcinoma at the American University of Beirut between 1990 and 2005 was conducted. Univariate and multivariate analyses were performed to determine the impact of surgery, chemotherapy, body mass index, bilirubin level and other factors on survival. RESULTS: The median survival of all patients was 8.57 mo (0.03-105.2). Univariate analysis showed that low bilirubin level (< 10 mg/dL), radical surgery and chemotherapy administration were significantly associated with better survival (P = 0.012, 0.038 and 0.038, respectively). In subgroup analysis on patients who had no surgery, chemotherapy administration prolonged median survival significantly (17.0 mo vs 3.5 mo, P = 0.001). Multivariate analysis identified only low bilirubin level < 10 mg/dL and chemotherapy administration as independent predictors associated with better survival (P < 0.05). CONCLUSION: Our data show that palliative and postoperative chemotherapy as well as a bilirubin level < 10 mg/dL are independent predictors of a significant increase in survival in patients with cholangiocarcinoma. PMID- 18506931 TI - Prevention and treatment of gastrointestinal dysfunction following severe burns: a summary of recent 30-year clinical experience. AB - AIM: To sum up the recent 30-year experience in the prevention and treatment of gastrointestinal dysfunction in severe burn patients, and propose practicable guidelines for the prevention and treatment of gastrointestinal (GI) dysfunction. METHODS: From 1980 to 2007, a total of 219 patients with large area and extraordinarily large area burns (LAB) were admitted, who were classified into three stages according the therapeutic protocols used at the time: Stage 1 from 1980 to 1989, stage 2 from 1990 to 1995, and stage 3 from 1996 to 2007. The occurrence and mortality of GI dysfunction in patients of the three stages were calculated and the main causes were analyzed. RESULTS: The occurrence of stress ulcer in patients with LAB was 8.6% in stage 1, which was significantly lower than that in stage 1 (P < 0.05). No massive hemorrhage from severe stress ulcer and enterogenic infections occurred in stages 2 and 3. The occurrence of abdominal distension and stress ulcer and the mortality in stage 3 patients with extraordinarily LAB was 7.1%, 21.4% and 28.5%, respectively, which were significantly lower than those in stage 1 patients (P < 0.05 or P < 0.01), and the occurrence of stress ulcer was also significantly lower than that in stage 2 patients (P < 0.05). CONCLUSION: Comprehensive fluid resuscitation, early excision of necrotic tissue, staged food ingestion, and administration of specific nutrients are essential strategies for preventing gastrointestinal complications and lowering mortality in severely burned patients. PMID- 18506932 TI - Venous diethylene glycol poisoning in patients with preexisting severe liver disease in China. AB - AIM: To analyze the clinical presentation of venous diethylene glycol (DEG) poisoning in patients with preexisting severe liver disease and factors that correlate with DEG poisoning. METHODS: Retrospective chart review was performed to analyze the epidemiology, clinical presentation, hepatorenal functions, hemodynamics and pathological characteristics of 64 patients with severe liver disease who received intravenous armillarisin-A, the solvent of which was DEG. Comparative analyses of correlating factors and causes for poisoning were based on the presence or absence of poisoning. RESULTS: Of the 64 patients who received armillarisin-A, 15 were found to have DEG poisoning. Twelve poisoned patients died. After a mean of 5 d, the poisoned patients displayed acute renal failure. Metabolic acidosis occurred in 13 cases. BUN, Cr, and CO2 values were significantly elevated and exacerbation of digestive tract symptoms and/or symptom was noted in 11 cases. Neurological system impairment was observed in 10 cases after 2 wk. Compared to the 49 non-poisoned patients, the poisoned patients exhibited significantly lower RBC and Hb values and higher WBC count. Renal biopsy from the poisoned patients revealed acute tubular necrosis and interstitial nephritis. Significant differences in preexisting severe hepatitis, ascites, renal disease, and diuretic therapy were found between groups. Prior to diethylene glycol injections, the mean values for neutral granular cells, BUN, Cr, calcium and phosphorous ions differed significantly between groups. CONCLUSION: Venous diethylene glycol poisoning is characterized by oliguric acute renal failure, metabolic acidosis, digestive symptoms, nervous system impairment, and a high probability of anemia and WBC proliferation. Mortality is high. Correlative factors include preexisting severe liver disease, renal disease, and infection. PMID- 18506933 TI - Effects of quercetin on hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol through the reactive oxygen species-nitric oxide pathway. AB - AIM: To investigate the effect of quercetin (3,3',4',5,7-pentahydroxy flavone), a major flavonoid in human diet, on hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol. METHODS: Forty male Sprague-Dawley rats, weighing 200-250 g, were randomly divided into control group (tap water ad libitum), ethanol treatment group (6 mL/L ethanol), quercetin treatment group (intragastric gavage with 100 mg/kg of quercetin per day), and ethanol plus quercetin treatment group (quercetin and 6 mL/L ethanol). Expression levels of proliferating cell nuclear antigen (PCNA) and Cyclin D1 were detected by Western blot to assay gastric mucosal cell proliferation in rats. To demonstrate the influence of quercetin on the production of extra-cellular reactive oxygen species/nitrogen species (ROS/RNS) in rats, changes in levels of thiobarbituric acid reactive substance (TBARS), protein carbonyl, nitrite and nitrate (NOx) and nitrotyrosine (NT) were determined. The activity of inducible nitric oxide synthase (NOS) including iNOS and nNOS was also detected by Western blot. RESULTS: Compared to control animals, cell proliferation in the gastric mucosa of animals subjected to ethanol treatment for 7 days was significant increased (increased to 290% for PCNA density P < 0.05, increased to 150 for Cyclin D1 density P < 0.05 and 21.6 +/- 0.8 vs 42.3 +/- 0.7 for PCNA positive cells per view field), accompanied by an increase in ROS generation (1.298 +/- 0.135 micromol vs 1.772 +/- 0.078 micromol for TBARS P < 0.05; 4.36 +/- 0.39 mmol vs 7.48 +/- 0.40 mmol for carbonyl contents P < 0.05) and decrease in NO generation (11.334 +/- 0.467 micromol vs 7.978 +/- 0.334 micromol P < 0.01 for NOx; 8.986 +/ 1.351 micromol vs 6.854 +/- 0.460 micromol for nitrotyrosine P < 0.01) and nNOS activity (decreased to 43% P<0.05). This function was abolished by the co administration of quercetin. CONCLUSION: The antioxidant action of quercetin relies, in part, on its ability to stimulate nNOS and enhance production of NO that would interact with endogenously produced reactive oxygen to inhibit hyper proliferation of gastric mucosal cells in rats treated with chronic oral ethanol. PMID- 18506934 TI - Proteasome inhibitor ameliorates severe acute pancreatitis and associated lung injury of rats. AB - AIM: To observe the effect of proteasome inhibitor MG-132 on severe acute pancreatitis (SAP) and associated lung injury of rats. METHODS: Male adult SD rats were randomly divided into SAP group, sham-operation group, and MG-132 treatment group. A model of SAP was established by injection of 5% sodium taurocholate into the biliary-pancreatic duct of rats. The MG-132 group was pretreated with 10 mg/kg MG-132 intraperitoneally (ip) 30 min before the induction of pancreatitis. The changes in serum amylase, myeloperoxidase (MPO) activity of pancreatic and pulmonary tissue were measured. The TNF-alpha level in pancreatic cytosolic fractions was assayed with an enzyme-linked immunosorbent assay (ELISA) kit. Meanwhile, the pathological changes in both pancreatic and pulmonary tissues were also observed. RESULTS: MG-132 significantly decreased serum amylase, pancreatic weight/body ratio, pancreatic TNF-alpha level, pancreatic and pulmonary MPO activity (P < 0.05). Histopathological examinations revealed that pancreatic and pulmonary samples from rats pretreated with MG-132 demonstrated milder edema, cellular damage, and inflammatory activity (P < 0.05). CONCLUSION: The proteasome inhibitor MG-132 shows a protective effect on severe acute pancreatitis and associated lung injury of rats. PMID- 18506936 TI - Multiple giant diverticula of the foregut causing upper gastrointestinal obstruction. AB - Small bowel diverticulosis represents an uncommon disorder (except for Meckel diverticulum) often misdiagnosed since it causes non-specific gastrointestinal symptoms. Most of times the diagnosis is carried out in case of related complications, such as diverticulitis, hemorrhage, perforation or obstruction. Intestinal obstruction can be caused by inflammatory stenosis due to repeated episodes of diverticulitis, volvulus, intussusception or jejunal stones. Herein we report a case of multiple jejunal diverticula causing chronic gastrointestinal obstruction. PMID- 18506935 TI - Change of intestinal mucosa barrier function in the progress of non-alcoholic steatohepatitis in rats. AB - AIM: To explore the change of intestinal mucosa barrier function in the progress of non-alcoholic steatohepatitis (NASH) in rats. METHODS: Thirty-two Sprague Dawley (SD) rats were randomly divided into control group and model group. Rats in the control group were given normal diet, and rats in the model group were given fat-rich diet. Eight rats in each group were killed at end of the 8th and 12th wk, respectively. The levels of endotoxin, D-xylose, TG, TC, ALT and AST, intestinal tissue SOD and MDA as well as intestinal mucus secretory IgA (sIgA) were measured. The pathology of liver was observed by HE staining. RESULTS: At end of the 8th wk, there was no marked difference in the levels of endotoxin, D xylose and sIgA between the two groups. At end of the 12th wk, rats in the model group developed steatohepatitis and had a higher serum level of endotoxin (P = 0.01) and D-xylose (P = 0.00) and a lower serum level of sIgA (P = 0.007). CONCLUSION: Intestinal mucosa barrier malfunction may exist in NASH rats and may be an important promoter of NASH in rats. PMID- 18506937 TI - Acute ischemic colitis during scuba diving: report of a unique case. AB - The presentation of clinical symptoms due to decompression during diving, varies significantly, as mainly minor disturbances for the gastrointestinal tract in particular have been reported. The following case debates whether diving can cause severe symptoms from the gastrointestinal system. We describe a clinical case of ischemic colitis presented in a 27-year-old male, who manifested abdominal pain while in the process of scuba diving 20 meters undersea, followed by bloody diarrhoea as soon as he ascended to sea level. Taking into account his past medical history, the thorough, impeccable clinical and laboratory examinations and presence of no other factors predisposing to ischemia of the colon, we assume that a possible relationship between diving conditions and the pathogenesis of ischemic colitis may exist. This unusual case might represent a hematologic manifestation of decompression sickness, due to increased coagulability and/or transient air emboli, occurring during a routine scuba diving ascent to sea level. PMID- 18506938 TI - A symptomatic cyst of the ligamentum teres of the liver: a case report. AB - Cysts of the liver ligaments are extremely rare and cysts of the ligamentum teres of the liver have been sporadically reported in the literature during the last century. The present report describes a case of a symptomatic patient with a cyst of the ligamentum teres of the liver. The patient presented with right upper quadrant pain and indigestion during the last 2 years. Ultrasound and computed tomography scans revealed a water-density mass attached to the anterior abdominal wall, but definite diagnosis could not be reached. The cyst was completely excised during laparotomy. Cysts of the ligamentum teres of the liver, although infrequent, may produce clinical symptoms and require excision. Ultrasound and computed tomography scan preoperatively cannot rule out malignancy, thus exploratory laparotomy and total resection of these lesions are necessary. PMID- 18506939 TI - Combined choriocarcinoma, neuroendocrine cell carcinoma and tubular adenocarcinoma in the stomach. AB - We described a patient with adenocarcinoma of the stomach combined with choriocarcinoma and neuroendocrine cell carcinoma. An 85-year-old man visited our hospital because of appetite loss. Gastric fiberscopy revealed a large tumor occupying the cardial region and anterior wall of the gastric body. The patient underwent total gastrectomy with lymphnode dissection and partial resection of the liver. Choriocarcinoma, small cell carcinoma and tubular adenocarcinoma existed in the gastric tumor. The choriocarcinomatous foci contained cells positive for beta-subunit of human chorionic gonadotropin (B-hCG) and human placental lactogen mainly in syncytiotrophoblastic cells. The small cell carcinomatous foci contained cells positive for synaptophysin, neuron-specific enolase (NSE), and chromogranin A. The prognosis for gastric adenocarcinoma with choriocarcinoma and neuroendocrine cell carcinoma is exceedingly poor. This patient died about 2 mo after the first complaint from hepatic failure. This is the first reported case of gastric cancer with these three pathological features. PMID- 18506940 TI - Pneumatosis cystoides intestinalis after fluorouracil chemotherapy for rectal cancer. AB - Pneumatosis cystoides intestinalis (PCI) is a relatively rare condition characterized by intraluminal gas in the gastrointestinal tract. Several chemotherapeutic agents have been reported to be associated with PCI, although fluorouracil-related PCI is extremely rare. We report a case of a 76-year old man who received adjuvant chemotherapy for rectal cancer with fluorouracil (FU) and leucovorin (LV). After 1 cycle of the treatment, he presented with diarrhea and abdominal pain. Abdominal radiogram revealed the presence of free air under the diaphragm and intramural gas in the intestine. Laparotomy was performed, showing a suspected diagnosis of perforation in the gastrointestinal tract. Intraoperative findings revealed penumatosis of the intestine without evidence of perforation. He was treated supportively and his symptoms improved. In conclusion, we should consider the possibility of PCI occurring in patients with malignancies during chemotherapy treatment. PMID- 18506941 TI - Diversity of citrus tristeza virus strains indicated by hybridization with cloned cDNA sequences. AB - Cloned cDNA sequences 500-2000 base pairs long, derived from a severe citrus tristeza virus (CTV) isolate, were used to study sequence homology with RNA of nine other serologically indistinguishable CTV isolates which differed in their biological properties. Six of the nine CTV isolates hybridized positively with the tested cDNA clones, while three others hybridized differentially with these cDNA clones. The potential use of cloned viral sequences for typing of virus strains is discussed. PMID- 18506942 TI - Molecular cloning of complementary DNA sequences of citrus tristeza virus RNA. AB - Complementary DNA (cDNA) of citrus tristeza virus (CTV) RNA, synthesized using calf thymus DNA random primers, was converted to a double-stranded form and inserted into the PstI site of the Escherichia coli pBR322 plasmid by the G-C tailing method. Bacterial clones harbouring virus-specific sequences were detected by colony hybridization with a 32P-labelled viral RNA probe. Hybridization patterns of denatured virus RNA revealed the presence of three types of specific clones: those hybridizing with a distinct narrow band corresponding to the full-length virus RNA, those hybridizing with a broader band of virus RNA sequences, and those hybridizing with several distinct virus-related RNA bands. Similar patterns were obtained when these clones were hybridized to purified double-stranded RNA from CTV-infected plants. None of these cDNA clones hybridized with similarly treated preparations extracted from healthy plants. The origin of variation among the CTV clones is discussed. PMID- 18506943 TI - Peter M. Black, M.D, Ph.D., F.A.C.S. PMID- 18506944 TI - [UNESCO's statements on race: chronicle of a lengthy endeavor, 1949-53]. PMID- 18506945 TI - [Ethnography of death in the Canary Islands at the start of the 20th century]. PMID- 18506947 TI - Death and mutilation at the dueling site: Pushkin's death as a national spectacle. PMID- 18506948 TI - [The escape of serfs as a sociopsychological phenomenon]. PMID- 18506949 TI - Mind, meaning and metaphor: the philosophy and psychology of metaphor in 19th century Germany. PMID- 18506950 TI - Editor's choice. Bare essentials. PMID- 18506951 TI - The paranormal Arctic: Lady Franklin, Sophia Cracroft, and Captain and "little weesy" Coppin. PMID- 18506952 TI - [Screening for dementia]. PMID- 18506953 TI - [Too old for organ donation?]. PMID- 18506954 TI - Continuity of patient care after hospital discharge. PMID- 18506956 TI - [Positive response to the new format]. PMID- 18506955 TI - Levosimendana. PMID- 18506957 TI - [Repercussions at the level of required health insurance of the introduction of three distinct professional titles in dentistry]. AB - From July 2007 on, Belgian dental graduates must complete one year of vocational training in general dentistry or a specialization in the field of either orthodontics or parodontology in order to obtain a licence for the Belgian Social Security System. This introduction of special professional titles required massive legislation in concert with State Health Insurance (RIZIV/INAMI), the Departments of Education of both Communities, manpower planning, comparable arrangements for physicians, and European Directives. Overall the process of subsequent changes in legislation and implementation took up to 15 years. The author includes some critical remarks and indicates what still needed to be done at the end of 2005 to conclude this far-reaching change in the organisation of dentistry in Belgium. PMID- 18506958 TI - [8 years of peer review in the context of accreditation]. AB - In order to maintain the quality of care, dental professionals must be enabled to update their knowledge on a regular basis. This can be achieved by continued education (lectures), but management techniques as well as regulations and legislation also have their influence. None of the aforementioned is equally effective or applicable. In 1998, peer review became a compulsory element of the system of accreditation for Belgian dentists. Peer review is an adaptation to the medical (dental) situation of what is known as a quality circle in management sciences. In contrast to former general opinion, today there is only little evidence of the effect of quality management on the quality level of (dental) care. Even continuous quality improvement experts admit that their expectations have not been met so far. The reason for this might be that quality management mainly focuses on side effects of care rather than on care itself; or that the strict methodology of the quality circle is violated. This article focuses at the methodology of the quality circle, compares with the medical literature and pays attention to the reasons why, at least in Belgium, peer review may not be very successful in improving quality of dental care. This is mainly due to mandatory participation at peer review within the Belgian accreditation system, and unfamiliarity with the methodology of peer review. In order to obtain quality improvement one must take integrated measures that effectively aim at multiple facets of a problem. If as many dentists as possible are to be involved, one has to guarantee a free choice between multiple alternatives in order to keep professional knowledge up to date. PMID- 18506959 TI - [Dental and buccal complications of lip and tongue piercing]. AB - Oral and facial piercing with different kinds of body art are being observed more frequently in medical and dental practices. Principally, piercing is not a new form of body art and is traditional in different geographical areas. In this review, the possible risks and complications are described. Postprocedural complications are oedema, haemorrhage and infection. Other adverse outcomes include mucosal or gingival trauma, chipped or fractured teeth, increased salivary flow, calculus build-up, and interference with speech, mastication and swallowing. Dentists, and oral- and maxillofacial surgeons should be in the position to advise patients with orofocial piercings or those who plan to have this type of body art performed. PMID- 18506960 TI - [The first issue of the Revue Belge de Medecine Dentaire in the new format]. PMID- 18506961 TI - [Radiologic exposure of the dental patient: comparison of the doses delivered by different techniques]. AB - This paper evaluates the doses delivered to the patient during several radiological procedures in dentistry: intraoral, panoramic and cephalometric radiography. Different digital techniques now available are compared to the AgBr film and film-screen technique. Absorbed doses at different organs are derived from measurements on dental radiological phantoms. The largest dose reductions are observed for intraoral radiography (31-84%). Significant dose reductions are also found for panoramic and cephalometric radiography (25-70% and 30-60%, respectively). By optimizing the exposure parameters and according to the ALARA principle, the smallest doses should be delivered to the patient that are needed to achieve the required quality of the images. Independently on the technique, the beam size should match as closely as possible the size of the detector. Collimation is particularly important for intraoral radiography. The dose at the thyroid should be kept as low as possible especially for children. For some beam incidences, a thyroid shield is especially efficient. The development of digital radiography and the related advantages should not lead to increasing the number of radiographs. The prescribed and performed types of examinations, and their number, should always be selected based on the clinical situation and on sound clinical judgment and experience in order to solve the raised medical problem. PMID- 18506962 TI - [Digital subtraction technique in dentistry]. AB - By computerized subtracting the grey values of geometrically corresponding pixels in consecutively taken digital X-ray images, it is possible to visualize or measure changes in radiodensity over time. This can be very useful, both in clinical and research applications, for the follow-up of tooth mineralization or bone changes. A major problem, however, resides in obtaining a sufficient geometrical standardization of the images. Several techniques have been proposed to this end, most of them still being in development, or too cumbersome to find application in daily routine. PMID- 18506963 TI - [Interview with Professor Reinhilde Jacobs. Interview by Th van Nuijs]. AB - Th. van Nuijs, chief editor of the Revue Belge de Medecine Dentaire, interviews professor Reinhilde Jacobs. R. Jacobs is professor of dento-maxillo-facial radiology and radioprotection at the Catholic University of Leuven. She is also head of the Oral Imaging Centre at the same university. PMID- 18506964 TI - [Gingival lichen planus]. AB - Lichen planus (LP) is a chronic inflammatory disease of uncertain etiology. When the gingiva is affected, the LP can appear under several clinical forms. The diagnosis of the gingival lichen planus is based on the anamnesis, the clinical observation and the histological analysis. Various medicamentous and non medicamentous treatments are used to treat gingival LP with random results, due to the lack of knowledge on the etiology and the recidiving character of the lesions. The risk of malignant transformation of gingival LP is weak but it depends on clinical forms, justifying a periodic follow-up of all patients. PMID- 18506966 TI - [Pain is a daily problem for the dentist]. PMID- 18506965 TI - [Multidisciplinary approach to behavior control of children during dental care: clinical study on the effect of learning by imitation]. AB - Many techniques of behavioral control of children have been developed to help the good performance of their health treatment. The objective of this study was to test the effect of the "modeling" technique on the pulse of the children during dental treatment. 69 girls and 86 boys were selected and divided into 3 groups A, B and C. Groups A and B were approached using the "modeling" technique and by taking respectively the mother or the father as a model. For the group C, the 'tell show do' approach was only used. The pulses of these children were registered during the check-up session and the cleaning therapy that followed. The results showed statistical significance (p < 0.001) of the two major factors investigated: gender of the children and choice of the model when using the modeling technique. PMID- 18506967 TI - [Physiopathology and clinical data of referred pain at the level of the masticatory system]. AB - In the differential diagnosis of orofacial pain, the importance of referred pain from other anatomical regions should be taken into account. This contribution describes the neuroanatomical and -physiological background of referred pain and the role of peripheral and central sensitization in its genesis and maintenance. In addition to the effects of afferent neurons, also the resulting efferent effects like increased muscle tone are illustrated, as well as the role of the autonomic nervous system. Through practical examples out of the daily dental practice, it is elucidated how referred pain can interfere in the diagnosis, and how the source and the location of pain can be discriminated. PMID- 18506968 TI - [Sensory injury to the lingual nerve after nerve block at the lingula mandibulae]. AB - Prolonged and possibly permanent change in sensation due to lingual nerve damage can occur after mandibular block anesthesia. The condition is rare and little can be done to prevent its occurrence. A case report is presented and functional as well as legal implications are discussed. Also practical recommendations are given to help the dentist when faced with the situation. PMID- 18506969 TI - [Desquamative gingivitis]. AB - The desquamative gingivitis is the clinical term given to the gingival manifestation of mucocutaneous diseases. It is characterized by an erythematous, glazed, friable and hemorrhagic gingiva, which can be accompanied by pains. Except the gingiva, the lesions can be localised on other oral mucous membranes or on the skin. The three principal diseases at the origin of the desquamative gingivitis are by order of frequency: the cicatricial pemphigoid, erosive lichen planus and pemphigus. The knowledge of the clinical, histological and immunohistochemical characteristics of these three affections is essential for the diagnosis and the suitable treatment of the desquamative gingivitis. PMID- 18506970 TI - [Toward a practical ethic]. AB - The relationship between ethics and philosophy and jurisdiction is described; different kinds of ethics are presented. The increasing pressure of liberal points of view has boosted the ethics of utility. The ethics of care oppose a too rational utilitarianism, taking into consideration relationships such as the caregiver-patient relationship. In the multicultural society ethics of care and virtue ethics are being criticised for not giving universal answers to ethical dilemmas. Can one still define "doing good"? Is "doing good" so culturally biased that it no longer provides the basis for ethical conduct? An accurate procedural assessment of values, sometimes interpreted quite differently in different cultures, could be a tool to judge values in a less relativistic way. PMID- 18506971 TI - [Red eye in children]. AB - Red eye in children is a common consultation purpose. Mostly benign, this sign may also cause visual impairment. We differentiate three kinds of red eye: localised, diffused and perikeratic injection. The last one must be recognized because of its association with severe ocular diseases. Diagnosis must be sure and treatment has to be efficient to not pertubate childrens visual development. Unfortunately, physical examination on children is not always easy. Consultation with an ophthalmologist is justified if a doubt remains, in case of chronic pathology or resistance to first intention treatment. PMID- 18506972 TI - [Adult depression. Improving screening and preventing recurrences]. PMID- 18506973 TI - [Epidemiology of mood disorders]. AB - The 12 months and lifetime prevalence of is respectively 5 and 9% in the general population; moreover 10 to 20% of general practice patients are depressed. Depression is involved in about 40 to 80% of suicide and induces one of the greatest social burden. Mood disorders are more frequent in women, individual living alone and people with low socio-economic level. Risk factors are stressfull life events, biological vulnerability (genetic factors), somatic diseases, psychiatric comorbidity including personality disorders and addictions. Depressive disorders are underdiagnosed and undertreated despite efficacious pharmacological and psychotherapeutic strategies in their treatment. It's the reason why public health programs to prevent depression have been promoted by many countries like European Alliance against Depression. Their efficacy must be evaluated as their cost-efficiency. PMID- 18506974 TI - [Adult depressive disorder: clinical aspects, nosography and psychiatrics comorbidity]. AB - Depressed mood and anhedonia are the main symptoms of depressive disorder. With other symptoms, it is responsible for a real break with premorbid patient's way of life. Several clinicals forms, caracterised by melancholia intensity, psychotic symptoms and cognitive troubles, must be identified to adapt treatment and to prevent patients from particular risks. The short and middle term prognosis is linked to the suicidal risk and the socioprofessionnal and familial consequences. Long term risk is the disease chronicisation, recurrence and bipolarity. The existence of other psychiatric comorbidity, such as alcoholism, personality disorder, anxiety disorder, is frequent and must be envisaged, despite the difficulty linked with symptoms intrication. PMID- 18506975 TI - [Depression and personality disorders]. AB - The association between depressive disorders and personality disorders is frequent. They need to be acknowledged in general practice for a better treatment (therapeutic alliance, compliance). Twenty to 50% of psychiatric inpatients and 50 to 85% of outpatients with a major depressive episode also have an associated personality disorder, most frequently borderline, avoidant, dependent or obsessive. These associations, as well as the so-called depressive personality disorder, have many implications for other persistant mood disorders such as dysthymic disorders. This article discusses the therapeutic options for an optimal treatment of these disorders. PMID- 18506976 TI - [Depression and physical illness]. AB - Depression associated to a physical illness may be under-recognized or under treated because of its diagnosis related difficulties and the trivialization of its presence in such a context. Now, this association contributes to physical illness' worsening, increased impairment, and may be responsible of care withdrawal, and even of suicide. The association between depression and physical illness may be a simple coincidence, or a psychological reaction to physical illness or else is the direct consequence of the somatic disease through biological disturbances or the iatrogenic effects of the disease treatment. It is also known, that depressive mood constitutes a significant risk factor for the development of various somatic disorders. Finally, a depression should be evoked when facing unexplained physical complaints. The entanglement between depression and physical illness needs to be particularly careful with prescriptions and especially an increased vigilance about drugs interactions. PMID- 18506977 TI - [Suicide risk and depression]. AB - Suicide occurs 20.4times more frequently in patients with major depression than in the general population. About 60% of the approximately 11,000 deaths from suicide each year in France occur in people with major depression. Assessment of suicide risk is one of the most challenging and important task for the clinician to continually perfect. According to the French consensus conference on the topic of the suicidal crisis, the assessment process includes the risk, the emergency and the dangerousness. The 8 steps of the crisis intervention are described. The strategies to ask how severe is the suicidal ideation is one of the most important point in an assessment interview. PMID- 18506978 TI - [Depression in elderly]. AB - Depression is frequent among aged persons and its diagnosis is difficult, because certain clinical presentations are unusual, explaining undertreatment. Depression is observed not only in ancient depressive people but can also be shown on elderly without previous mood disorders. Somatic symptoms often mask depression. More over, somatic diseases can interfere with depression. A hypochondria is always associated with depression in old patients, patients expressing in this a particular way a psychological pain. Loss of motivation is sometime a depression in aged people, associated with a dysexecutive syndrome or, elsewhere, aggressiveness can reveals a hostile depression. At a worst stage of the disease, in particular, when a dementia is latent, confusion is present. A therapeutic test with an antidepressant can help to discriminate between a dementia with depressive symptoms or a depression with a confusion. But, even if the cognitive disorders disappear with the treatment, a dementia can appear years after. PMID- 18506979 TI - [Rationalizing the prescription of antidepressants]. AB - The prescription of antidepressant drugs should only concern depressed patients fulfilling diagnostic criteria for a major depressive episode. This prescription should remain as simple as possible and the association of other psychotropic drugs should not be systematic. Optimal doses should be reached rapidly but the therapeutic efficacy will only become obvious after 2-4 weeks and complete after 6-8 weeks. As most antidepressants display a similar therapeutic efficacy, the choice will depend on tolerability, patient characteristics and the result of previous therapeutic attempts. Prescribing an antidepressant requires a thorough supervision and repeated visits during the first weeks of treatment. The recommended duration of treatment for a depressive episode is six months and the medication should he tapered off progressively in order to avoid withdrawal symptoms. PMID- 18506980 TI - [Behavioural and cognitive therapies in depression]. AB - A large number of psychotherapies were proposed to depressed patients. The behavioural and cognitive therapies (TCC) showed their efficacy. This short term therapy improves quickly the symptoms and prevents around 30% of relapse. The TCC works on the depressed symptoms (engaging the patients in a large number of pleasant activities), on hopelessness cognitions and on the own psychological vulnerability (named cognitive schemata). The activation of depressed schemata is correlated with relapse). The indication of TCC in depressed patient is discussed after two or three sessions performed by a cognitive and behaviour therapist. This professional has preliminary learnt the theory and practice during a specific training. PMID- 18506981 TI - [Anxiety and depression. Self-questionnaire HAD (Hospital Anxiety and Depression Scale)]. PMID- 18506982 TI - [Governance, paradigm, metrology and benchmarking!]. PMID- 18506983 TI - [National ranking exam. N63. What could fall at the exam?]. PMID- 18506984 TI - [Breast tumours]. PMID- 18506985 TI - [Psychosis and chronic delirium]. PMID- 18506986 TI - [Lung tumours, primary and secondary]. PMID- 18506987 TI - [Influenza]. PMID- 18506988 TI - [Pyoderma gangrenosum is 100 years old. From Louis Brocq to biotherapies]. PMID- 18506989 TI - [Bisphosphonates]. PMID- 18506990 TI - [Interview with Professor Eric Fossion on the subject of bisphosphonates. Interview by Th van Nuijs]. AB - Th. van Nuijs, chief editor of the Revue Belge de Medecine Dentaire, interviews Professor Eric Fossion on the problematic of bisphosfonates and their role in the osteonecrosis of the jaws. Prof. Eric Fossion is maxillo-facial surgeon. Actually, he is professor part-time at the Catholic University of Leuven. He is Head Department of Dentistry, Oral and Maxillofacial Surgery, Middelheim Hospital, Antwerp. PMID- 18506991 TI - [Osteonecrosis of the jaws: a secondary effect of bisphosphonates]. AB - When treated with intravenous bisphosphonates to prevent bone resorption, 4-10% of the patients may develop osteonecrosis of the jaws a side effect. The osteonecrosis is usually preceded by an invasive dental procedure such as the removal of a tooth. Treatment of the osteonecrosis should be conservative using aseptic mouth rinses, oral antibiotics and small debridement. It seems advisable to have a dental check-up before starting using intravenous bisphosphonates, since a potential dental infections can be treated before the bone healing capacity is reduced by the bisphosphonates. PMID- 18506992 TI - [Subgingival application of chlorhexidine in the treatment of periodontitis]. AB - The clinician is frequently faced with poor treatment outcome of scaling and root planing predominantly in deep pockets and furcations. In an attempt to optimize clinical effects at these weakly responding sites chemo-mechanical treatment strategies have been developed and studied, among those using chlorhexidine as an antimicrobial agent. In this overview the clinical effects of subgingival chlorhexidine irrigation, gel, chip and varnish administration as an adjunct to scaling and root planing in the treatment of periodontitis are discussed. There is little to no evidence indicating treatment outcome will benefit from subgingival irrigation using chlorhexidine solutions. Furthermore, the use of a chlorhexidine solution as a coolant during (ultra)sonic scaling does not result in a clinically superior response when compared to water cooling. Despite the high viscosity of a gel, its use cannot be justified in the treatment of periodontitis as clinical effects are also negligible. Multi-center studies have indicated significantly higher pocket reductions and clinical attachment gains following a combination of scaling and root planing and the administration of a chlorhexidine chip. As some studies failed to confirm this finding, more research is needed to elucidate the additional value of the chip. The subgingival administration of a supersaturated chlorhexidine varnish seems promising as additive clinical effects have been reported. Large-scale studies are however lacking on this treatment concept. The clinician should continue treating periodontitis primarily by mechanical means. Supplemental chemical therapy based on subgingival chlorhexidine administration can be considered; yet, solutions and gels are not suitable as compensatory aids when scaling and root planing becomes less effective. PMID- 18506993 TI - [Amalgam, composites and compomers: comparative histologic study of effects on the periodontal tissues]. AB - Amalgam, composite resin and compomer are frequently used in operative dentistry today. Earlier studies have shown the periodontal consequences induced by these materials if they have intracrevicular restorative margins. This study was performed on dogs where Class V cavities with apical margin located at the alveolar bone crest were prepared and restored with these materials. Three months later, the experimental and control sites were prepared for histological analysis. Mean bone loss was significant only with compomer. The mean length of the supracrestal connective tissue was almost the same with the three fillings. The length of epithelial attachment was the highest with composites. There was a decrease in the length of the biologic width from Day 0 to Day 90. PMID- 18506994 TI - Boron nanoparticles inhibit tumour growth by boron neutron capture therapy in the murine B16-OVA model. AB - BACKGROUND: Boron neutron capture therapy usually relies on soluble, rather than particulate, boron compounds. This study evaluated the use of a novel boron nanoparticle for boron neutron capture therapy. MATERIALS AND METHODS: Two hundred and fifty thousand B16-OVA tumour cells, pre-incubated with boron nanoparticles for 12 hours, were injected subcutaneously into C57BL/6J mice. The tumour sites were exposed to different doses of neutron radiation one, four, or eight days after tumour cell inoculation. RESULTS: When the tumour site was irradiated with thermal neutrons one day after injection, tumour growth was delayed and the treated mice survived longer than untreated controls (median survival time 20 days (N = 8) compared with 10 days (N = 7) for untreated mice). CONCLUSION: Boron nanoparticles significantly delay the growth of an aggressive B16-OVA tumour in vivo by boron neutron capture therapy. PMID- 18506995 TI - A new micronized formulation of 2-methoxyestradiol-bis-sulfamate (STX140) is therapeutically potent against breast cancer. AB - There is a continued need for orally bioavailable anticancer compounds that exhibit good efficacy against breast cancer. STX140, a derivative of 2 methoxyestradiol (2-MeOE2), has been shown to have excellent oral bioavailability and significantly reduces tumor growth. A new micronized formulation of STX140 has now been developed and its pharmacokinetics (PK) in rats and effect on MDA-MB 231 breast cancer growth in nude mice was investigated. MATERIALS AND METHODS: For the PK studies, female Wistar rats were treated orally with STX140 in two separate vehicles (10% tetrahydrofuran (THF) in propylene glycol (PG) or 0.5% methyl cellulose (MC) in saline) and plasma samples taken for high performance liquid chromatography analysis over 48 h. For the tumor efficacy studies, female nude mice were inoculated with MDA-MB-231 breast cancer cells and then treated orally with a range of doses of STX140. RESULTS: The PK studies demonstrated that the THF/PG vehicle resulted in a greater oral bioavailability of STX140 compared to the 0.5% MC vehicle. However, this was not translated to the tumor efficacy studies where STX140 at 20 mg/kg in either vehicle caused a significant reduction in tumor volume. CONCLUSION: The new micronized formulation of STX140 is orally bioavailable and efficacious at inhibiting MDA-MB-231 breast tumor growth. PMID- 18506996 TI - Cellular response to irinotecan in colon cancer cell lines showing differential response to 5-fluorouracil. AB - BACKGROUND: Use of irinotecan (CPT-11) as second-line therapy for metastatic colorectal cancer has shown some promise in cases where 5-fluorouracil (5-FU) has failed. Cross-resistance to both drugs may however be a potential clinical problem. The cellular response to CPT-11 was investigated in two human colon cancer cell lines that demonstrate a differential response to 5-FU. MATERIALS AND METHODS: Cell cycle progression, clonogenic survival, DNA damage checkpoint activation, apoptosis induction and senescence development were assessed during 48 hours of treatment and 72 hours of recovery. RESULTS: Both cell lines had similar cellular response patterns to CPT-11. Growth inhibition, loss of clonogenicity, ataxia telangiectasia mutated (ATM) activation, H2AX phosphorylation, TP53 stabilization, CDKN1A induction, G2/M arrests, endoreduplication, negligible cell death and appearance of a senescence associated beta-galactosidase phenotype were observed. CONCLUSION: Cross resistance to 5-FU and CPT-11 was not demonstrated. The appearance of a senescence phenotype in response to CPT-11 treatment may have potential clinical relevance for treatment regimens. PMID- 18506997 TI - Acquired resistance to Fas/CD95 ligation in U937 cells is associated with multiple molecular mechanisms. AB - BACKGROUND: Acquired resistance to apoptosis is a critical event in tumour development and in insensitivity toward therapy. To investigate resistance mechanisms to Fas/CD95/Apo-1-induced apoptosis, a Fas ligand-resistant variant of the U937 cell line was generated. RESULTS: Selection for Fas resistance resulted in a partial cross-resistance to TRAIL and TNF-alpha. Activation of caspase-8 was found to be impaired and the expression of Fas was reduced. However, FADD expression and ligand-induced aggregation of Fas was intact. Inhibition of various signalling pathways with pharmacological inhibitors revealed that resistance to death receptor-mediated apoptosis was dependent on altered tyrosine phosphatase/kinase activities and de novo protein synthesis. Moreover, FLIP, an anti-apoptotic protein, was expressed to a higher extent in the resistant cells. CONCLUSION: We provide evidence that acquired resistance to Fas-induced apoptosis in U937 cells involves a discrete set of molecular mechanisms which also render the cells cross-resistant to other death ligands. PMID- 18506998 TI - Adenovirus-mediated inhibitor kappaB gene transfer improves the chemosensitivity to anticancer drugs in human lung cancer in vitro and in vivo. AB - BACKGROUND: Nuclear factor kappaB (NFkappaB) is a transcription factor which is importantly implicated in cancer cell growth. In a previous report, we confirmed that lung cancer cell growth was suppressed significantly by the blockade of NFkappaB function. In this study the combination effect of chemotherapy and inhibition of NFkappaB on the human lung cancer cell line, NCI-H460, in vitro and in vivo was investigated. MATERIALS AND METHODS: In the in vitro experiment, 50% of cell growth inhibitory concentrations (IC50) of chemotherapy agents were determined alone or when combined with adenovirus mediated IkappaBalpha gene transfer. Annexin-V/PI stain and caspase 3 activity measurement were used to detect the apoptosis caused by treatment. In the in vivo experiment, the tumor growth suppressive effect of combination treatment was evaluated for tumor bearing mice. NFkappaB, p53 and VEGF expression in the tumors were also analyzed immunohistologically. RESULTS: Several chemotherapy agents, including paclitaxel, showed lower IC50s when combined with AdIkappaBalpha infection in vitro. Apoptosis through activation of the caspase 3 pathway was enhanced by the combination treatment. For established NCI-H460 tumors, combined treatment significantly inhibited tumor growth. Immunohistochemical staining showed increased expression of p65 after paclitaxel treatment, while paclitaxel in combination with AdIkappaBalpha intratumoral injection eliminated this expression accompanied by the slightly reduced expression of VEGF, with stable p53 status. CONCLUSION: A combination of chemotherapy and IkappaBalpha could inhibit tumor growth effectively by blocking the expression of NFkappaB and inducing apoptosis. Moreover, it might allow reduction of the dose of chemotherapy agents and provide benefit for clinical application. PMID- 18506999 TI - Apoptosis induced by an alkylated purine, 6-dimethylaminopurine, and changes in gene expression in human lymphoma U937 cells. AB - Apoptosis induced by an alkylated purine, 6-dimethylaminopurine (6-DMAP), was investigated to explore the p53-independent pathway in a human lymphoma U937 cell line. Here, it was discovered that the apoptosis was induced by 6-DMAP in a dose- and time-dependent manner and treatment for 16 h at a concentration of 5 mM induced apparent DNA fragmentation, phosphatidylserine externalization and lowering of the mitochondrial membrane potential, which are typical markers of apoptosis. Western blotting revealed reduced expression in Bcl-XL, increased expression in Bax and release of cytochrome c. These were associated with activation of caspase-3. The 6-DMAP-induced apoptosis was preceded by an increase in the intracellular calcium ion concentration ([Ca2+]i), indicating the involvement of intracellular Ca2+ ions in apoptosis. Two independent sets of cDNA microarray systems were used to examine changes in gene expression. Of the 3,893 and 886 genes analyzed, 32 and 13 genes were identified as down-regulated in cells treated with 6-DMAP for 3 h. No up-regulated gene was found. Real-time PCR revealed a significant decrease in mRNA of proliferating cell nuclear antigen, insulin-induced gene 1, serine proteinase inhibitor 2 and v-myc. Pathway analysis also revealed the interaction of genes on down-regulation. These findings suggest that intracellular calcium ions and mitochondrial caspase-dependent pathways play major roles in 6-DMAP-induced apoptosis, and protein kinase inhibition by the agent causes massive down-regulation of all genes relating to cell proliferation and progression of the cell cycle to induce apoptosis. PMID- 18507000 TI - Antisense-Snail transfer inhibits tumor metastasis by inducing E-cadherin expression. AB - Carcinoma cell metastasis is mediated by increased mobility and invasiveness via epithelial-mesenchymal transition (EMT). Snail, a zinc-finger transcription factor, triggers EMT by directly repressing E-cadherin expression. In this study, the correlation between E-cadherin and Snail expression was investigated. In addition, the effect of antisense-Snail on the mobility and invasive properties of tumor cells in vitro and in vivo was assessed. Our data showed that Snail induced EMT, up-regulated mesenchymal gene expression and increased the ability of cancer cells to migrate and invade. In contrast, antisense-Snail prevented EMT by decreasing the expression of mesenchymal genes, and inhibited the migration and invasive potential in vitro. An in situ xenograft model showed that adenovirus-mediated antisense-Snail expression inhibited tumor cell metastasis to the lymph nodes and prolonged the survival of the hosts. These findings suggest that Snail plays a critical role in EMT and subsequent invasiveness. Snail inhibition may serve as a promising target for therapeutic strategies to prevent tumor metastasis. PMID- 18507001 TI - Combination therapy of S-1 with selective cyclooxygenase-2 inhibitor for liver metastasis of colorectal carcinoma. AB - BACKGROUND: Hepatic resection, the only curative treatment for liver metastasis of colorectal cancer has become standard treatment, but most cases of liver metastases are inoperable, and of the patients treated with hepatectomy about 50% have a recurrence in the liver. The aim of this study was to establish preventive therapy for the liver metastasis of colorectal cancer. MATERIALS AND METHODS: In this study, a combined treatment with S-1 and a selective COX-2 inhibitor for liver metastasis of colorectal cancer was developed. The effect of these agents on the proliferation and invasion of a highly metastatic human colon cancer cell line, LM-H3, was examined. RESULTS: 5-Fluorouracil (5-FU) had an inhibitory effect on the proliferation of LM-H3 cells, but no inhibitory effect on the invasion of LM-H3 cells in in vitro experiments. 5-Chloro-2,4-dihydroxypyridine (CDHP) had no antitumor activity itself, but the inhibitory effect of 5-FU on the proliferation was enhanced by adding CDHP. COX-2 inhibitors, etodolac and rofecoxib, did not have an inhibitory effect on the proliferation of LM-H3 cells at low concentrations, but had significant inhibitory effect on the invasion of LM-H3 cells in in vitro experiments. In a nude mouse liver metastasis model, combined treatment with S-1 and a COX-2 inhibitor more effectively restrained liver metastasis of LM-H3 cells than either alone. This outcome was most likely due to S-1 inhibiting proliferation of and the COX-2 inhibitor inhibiting invasion of LM-H3 cells. CONCLUSION: Combined therapy with S-1 and a COX-2 inhibitor might hold promise for prophylaxis of liver metastasis of colorectal cancer. PMID- 18507002 TI - DNA damage response in prostate cancer cells after high-intensity focused ultrasound (HIFU) treatment. AB - BACKGROUND: High-intensity focused ultrasound (HIFU) is a new modality that can be used for local tumor ablation therapy. In the present study, the expression of human checkpoint kinase2 (Chk2), which may have significant roles in the response to DNA damage after HIFU treatment for prostate cancer, was examined. MATERIALS AND METHODS: By immunohistochemistry, the Chk2 expression in human prostate cancer tissues was examined before and after HIFU treatment. The phosphorylation of ataxia-telangiectasia-mutated kinase (ATM), histone H2A variant (H2AX), Chk2 and p53, and the number of apoptotic cells in human prostate cancer cell lines after heat treatment was checked. RESULTS: The expression level of phosphorylated Chk2 was found to be increased after HIFU treatment. In addition, heat treatment induced the phosphorylation of Chk2 proteins examined and led to apoptosis in the prostate cancer cells. CONCLUSION: Our results suggest that DNA double-strand break formation is a possible pathway of HIFU treatment and leads to heat-induced cell killing. PMID- 18507003 TI - Identification of novel genes involved in the synergistic antitumor effect of caffeine in osteosarcoma cells using cDNA macroarray. AB - BACKGROUND: Caffeine enhances the cytocidal effects of DNA-damaging agents. This study investigated genes involved in the synergistic effect of caffeine on osteosarcoma cells using gene-profiling analysis. MATERIALS AND METHODS: Sensitivity to cisplatin and the synergistic effect of caffeine were evaluated in five osteosarcoma cell lines with different p53 gene status. Gene expression profiles were analyzed using cDNA macroarray and verified by real-time RT-PCR. RESULTS: The cell lines were grouped into three types with different cytotoxic patterns. Comparison of profiling data from these groups identified twelve novel genes associated with the synergistic effect of caffeine. Real-time RT-PCR analyses verified up-regulation of two apoptosis-enhancing genes and down regulation of two interferon-inducible genes related to the synergy of caffeine. CONCLUSION: These findings provide new insights into the molecular mechanisms of the synergistic effect of caffeine related to p53 gene status in osteosarcoma, providing candidates for an assay of responsiveness to caffeine-potentiated chemotherapy for osteosarcoma. PMID- 18507004 TI - Down-regulation of plakoglobin in soft tissue sarcoma is associated with a higher risk of pulmonary metastasis. AB - Soft tissue sarcomas (STS) behave with aggressiveness and metastatic potential, that can vary depending on their locations. There has been little information on the exact molecular mechanisms involved in their biological aggressiveness. To identify genes involved in the differences, the gene expression profiles were compared between STS-orthotopic and heterotopic implanted models, and their significance in human STS was verified. Human fibrosarcoma HT1080 cells were implanted either in the quadriceps femoris muscles or footpads of nude mice, and the gene expression profiles of the tumors were compared by cDNA arrays. The mRNA and protein levels of the identified genes were examined by both real time RT-PCR and immunohistochemistry not only in the tumors of the models, but also in clinical STS. The implanted HT1080 cells demonstrated different growth and metastatic potentials depending on their implant locations. cDNA array analyses showed decreased expression of the plakoglobin gene in the intramuscle-implanted group, which was statistically confirmed by real-time RT-PCR (p = 0.04). Plakoglobin was immunolocalized diffusely in the cytoplasm of tumor cells implanted in the footpads, but not those in the muscle. Real-time RT-PCR assays of clinical STS showed that the mean plakoglobin/glyceraldehyde 3-phosphate dehydrogenase (G3PDH) ratio in primary sarcoma tissues with pulmonary metastases (0.92) was significantly lower than in those without metastasis (6.58) (p < 0.0001), and that STS cases with high plakoglobin gene expression had an excellent prognosis. These results suggest that plakoglobin gene expression level might be useful as a new biomarker for metastasis and prognosis of human STS. PMID- 18507005 TI - Role of histone deacetylase inhibitor in adenovirus-mediated p53 gene therapy in esophageal cancer. AB - BACKGROUND: Recently, the use of histone deacetylase inhibitors (HDACI) with gene therapy has been shown to improve the effect of this therapy. The effectiveness of one of the novel HDACIs, FK228, was examined in adenovirus-mediated p53 gene therapy of esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: The expression levels of coxsackie and adenovirus receptor (CAR) in ESCC patients were examined immunohistochemically. CAR induction by FK228 in ESCC cells was analyzed by real-time PCR and Western blotting. The efficiencies of adenoviral transduction treated with FK228 were determined using AV1.0CMV-betagal. The acetylation of p53 protein was detected by Western blotting. RESULTS: CAR expression was reduced in some tumor specimens compared to that in normal specimens. CAR expression was increased by FK228 in both in vitro and in vivo experiments. FK228 improved the efficiency of adenovirus infection. Acetylated p53 protein was increased in a dose- and a time-dependent manner. CONCLUSION: Our findings suggest that FK228 has a potent ability to augment the effect of adenovirus-mediated p53 gene therapy in ESCC cells. PMID- 18507006 TI - Identification of potential chemoresistance genes in osteosarcoma. AB - BACKGROUND: Osteosarcoma (OS) is an aggressive bone malignancy that primarily affects children and adolescents. Patients with metastatic disease at diagnosis have only a 20% survival rate. The poor survival rate of these patients is largely due to their lack of responsiveness to chemotherapy. However, the mechanisms underlying osteosarcoma chemoresistance remain unknown. MATERIALS AND METHODS: The effect of cisplatin, doxorubicin and etoposide was examined on OS cell lines. Affymetric Genechip analysis was used to examine differential gene expression. RESULTS: A correlation between increasing metastatic potential and increasing chemoresistance was observed in the MG-63 cell line and sub-line model. Microarray analysis of these cell lines revealed the differential expression of several genes potentially involved in chemoresistance including ABCG2, ADD3, NMT2, WNTSa and PTN. CONCLUSION: The identification of genes contributing to chemoresistance and determining the role these genes play is critical in characterizing patient responsiveness and overcoming chemoresistance in osteosarcoma patients. PMID- 18507007 TI - Implications of the involvement of the endoplasmic reticulum stress pathway in drug-induced apoptosis. AB - Apoptosis occurs by distinct pathways that involve the cell surface, mitochondria or the endoplasmic reticulum. Previous studies had shown that deoxyadenosine resistant L1210 cells (Y8) proceeded to apoptosis under conditions in which the parental L1210 cell line (WT) did not undergo an apoptotic response. Combinations of drugs, acting at different molecular targets, markedly potentiated the apoptotic response in the Y8 cells without inducing apoptosis in the WT cells. In the present study, induction of apoptosis by parthenolide and BAY 11-7085, drugs that targeted nuclear factor kappa B activation, was blocked by the presence of N acetylcysteine (NAC). On the other hand, the levels of apoptosis induced by parthenolide or BAY 11-7085 were increased by pre-treatment of the cells with glutathione lowering L-buthionine-(S,R)-sulfoximine (BSO). Western blot analyses showed that the levels of the stress proteins, Grp 78 and Gadd 153 were reduced in the parthenolide-treated Y8 cells, but not in those co-treated with NAC. Protection of the cells from apoptosis induced by parthenolide or BAY 11-7085 by NAC was relatively specific as the induction of apoptosis in the Y8 cells by MG 132, flavopiridol, Gemcitabine or PRIMA-1 was not decreased by NAC. These data suggest that multiple pathways, one of which is ER-stress induced, may ultimately be involved and interactive in the induction of apoptosis in specific cell lines. PMID- 18507008 TI - Effect of bortezomib on EGFR expression in head and neck squamous cell carcinoma cell lines. AB - BACKGROUND: Overexpression of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinoma (HNSCC) has often been correlated with poor prognosis. Recent investigations have shown that the proteasome inhibitor bortezomib exhibits a high antiproliferative and apoptotic activity in HNSCC cell lines. The present study investigated whether bortezomib has an effect on EGFR expression in different squamous cell carcinoma cell lines. MATERIALS AND METHODS: Six EGF-stimulated or non-stimulated squamous carcinoma cell lines were treated with bortezomib. Western blots were performed to determine EGFR expression. For statistical analysis, a Wilcoxon test for matched pairs (dependent samples) was performed using SPSS 13.0 software for Windows. RESULTS: Changes in EGFR expression after bortezomib treatment in EGF non-stimulated and EGF-stimulated squamous carcinoma cell lines failed to reach statistical significance in either experimental group (p > 0.05). CONCLUSION: Given the high expression of EGFR in head and neck tumors, further investigations should address the question whether the apoptotic activity of bortezomib can be enhanced by adding an anti-EGFR antibody. PMID- 18507009 TI - Intraperitoneal infusion of recombinant plasminogen activator inhibitor type 2 induced apoptosis in implanted human colon cancer and inhibited its growth and liver metastasis. AB - Antitumor effects of plasminogen activator (PA) inhibitors (PAls) were analyzed in a mouse model of human colon cancer xenografts. Either recombinant PA inhibitor-1 (rPAI-1) or inhibitor-2 (rPAI-2) was injected intraperitoneally to nude mice bearing human colon cancer xenografts for 6 weeks. Primary tumors in rPAI-2-treated group were smaller (0.45 +/- 0.13 g, n = 16) than in the other two groups (control: 0.73 +/- 0.24 g, n = 15; rPAI-1: 0.62 +/- 0.29 g, n = 19). Primary tumors in the rPAI-2-treated group exhibited less mature ductal structures and were significantly smaller. The apoptotic index was higher in the rPAI-2-treated group (4.64 +/- 2.12%) than in the other groups (control: 1.94 +/- 0.82%; rPAI-1: 2.08 +/- 1.07%). Liver metastasis was less frequent in the rPAI-1 (5/19) and rPAI-2-treated groups (1/16) than in the control group (14/15). PAI-2 more effectively suppressed tumor metastasis and progression, probably by inducing apoptosis; some different unknown mechanism may cause the difference in both antitumor effect and the histological findings. This may indicate the therapeutic potential of these PAls in malignant patients. PMID- 18507010 TI - Comparative anti-inflammatory activities of curcumin and tetrahydrocurcumin based on the phenolic O-H bond dissociation enthalpy, ionization potential and quantum chemical descriptor. AB - Curcumin and its reduced derivative tetrahydrocurcumin have been shown to exhibit chemopreventive activity. Cyclooxygenase-2 (COX-2) inhibition in lipopolysaccharide (LPS)- or Porphyromonas gingivalis fimbria-stimulated RAW 264.7 cells was investigated using Northern blot analysis. The fimbria-stimulated expression of the COX-2 gene was inhibited by curcumin but not by tetrahydrocurcumin. LPS-stimulated COX-2 gene expression was completely inhibited by curcumin, but an increase in the concentration of tetrahydrocurcumin did not cause complete inhibition of COX-2 expression. The inhibitory effect of curcumin on nuclear factor kappa B (NF-kappaB) activation in the cells was clearly observed, but that of tetrahydrocurcumin was incomplete even at a concentration of 20 microM. To explain the difference in effect between the two compounds, analysis of the frontier orbital was performed using ab initio 6-31G* wave function. The calculated chemical hardness (eta) for curcumin was clearly smaller, whereas its electronegativity (chi) and electrophilicity (omega) were clearly greater than the corresponding values for the curcumin-related compounds tetrahydrocurcumin, isoeugenol and eugenol. This suggested that the anti inflammatory activities of curcumin may be related to eta-, chi- and/or omega controlled enzymes. In addition, the bond dissociation enthalpy (BDE) of the phenolic OH was calculated using the density function theory (DFT)/B3LY. The total BDE values of curcumin and tetrahydrocurcumin were almost identical, but the BDE of one-electron oxidation and ionization potential (IP) for curcumin were lower than those for tetrahydrocurcumin, suggesting the highly pro-oxidative activity of curcumin. Curcumin has both oxidant and antioxidant properties. A causal link between the anti-inflammatory activities and molecular properties of phenolic antioxidants is suggested. PMID- 18507011 TI - Involvement of the estrogen receptor beta in genistein-induced expression of p21(waf1/cip1) in PC-3 prostate cancer cells. AB - BACKGROUND: Dietary genistein, a phytoestrogen derived from soybean, has been suggested as a chemopreventive agent for prostate cancer. Genistein has been reported to exert its anticancer effects via a variety of functional pathways, but the upstream signaling of molecules regulated by genistein remains unclear. In this study, estrogen receptor (ER) beta involvement in genistein-induced expression of cell cycle inhibitors in PC-3 prostate cancer cells was investigated. MATERIALS AND METHODS: The proliferation of PC-3 cells exposed to genistein was measured by the water-soluble tetrazolium salt (WST-1) proliferation assay. The expression of p21, p27 and ERbeta in the PC-3 cells was assessed by quantitative real-time reverse transcription-PCR. ERbeta silencing was performed using a small interfering RNA (siRNA). The transcriptional activity of the p21 promoter was determined by the luciferase reporter assay. RESULTS: Genistein caused marked inhibition of proliferative activity and induced the expression of p21 and ERbeta in the PC-3 cells. The siRNA against ERbeta suppressed the genistein-induced expression of p21 and reduced the transactivation activity of the p21 promoter induced by genistein. CONCLUSION: ERbeta is involved in genistein-induced expression of p21 in PC-3 cells. PMID- 18507012 TI - Changes to N-linked oligosaccharide chains of human serum immunoglobulin G and matrix metalloproteinase-2 with cancer progression. AB - BACKGROUND: Alterations to the sugar chain structure of E-cadherin, a calcium dependent adhesion molecule, have been shown to influence cancer metastasis. Furthermore, expression of sialyl Le(x) sugar chains on cancer cells has been demonstrated to influence their adhesion to vascular endothelial cells. On the other hand, matrix metalloproteinase-2 (MMP-2) degrades extracellular matrix and is involved in the invasion and metastasis of cancer cells. PATIENTS AND METHODS: N-linked oligosaccharides of human serum immunoglobulin G (IgG) were analyzed in 36 patients with localized or metastatic cancer (12 lung, 12 gastric and 12 prostate cancer) and 10 healthy controls using fluorophore-associated carbohydrate electrophoresis (FACE). MMP-2 levels in the sera were determined by enzyme immunoassay. RESULTS: Fr1 (monogalactosyl IgG oligosaccharide) and Fr2 (digalactosyl IgG oligosaccharides) were significantly decreased (p < 0.001), while Fr4 (agalactosyl IgG oligosaccharides) were significantly increased (p < 0.001) with cancer metastasis. The Fr4/Fr1+Fr2 ratio in localized and metastatic cancer was significantly increased compared to healthy controls (p < 0.001), and was significantly higher in metastatic than localized cancer (p < 0.001). Serum MMP-2 levels in metastatic cancer were significantly higher than in localized cancer (p < 0.001). There was a good correlation between the Fr4/Fr1+Fr2 ratio and serum MMP-2 levels in patients with metastatic cancer (p < 0.0001). CONCLUSION: The analysis of serum IgG N-linked oligosaccharide chain structures by FACE may be an auxiliary indicator of serum tumor markers useful for monitoring cancer progression. PMID- 18507013 TI - Transforming growth factor-beta1 modulates tumor-stromal cell interactions of prostate cancer through insulin-like growth factor-I. AB - Insulin-like growth factor-I (IGF-I) secreted from the prostate stroma mediates tumor-stromal cell interactions in prostate cancer development. We have recently reported that human prostate stromal cells (PrSC) stimulate human prostate cancer DU-145 cell growth via IGF-I. Transforming growth factor-beta1 (TGF-beta1) also plays a critical role in tumor-stromal cell interactions, but TGF-beta1 has pleiotropic effects as it can modulate growth of prostate cancer either positively or negatively. To elucidate the complex behavior of TGF-beta1, the effect of TGF-beta1 on the IGF axis in PrSC was studied. TGF-beta1 increased the expression of IGF-I and IGF binding protein-3 (IGFBP-3). RT-PCR experiments revealed that TGF-beta1 upregulated the mRNA expression of IGF-I and IGFBP-3. However, while TGF-beta1 significantly increased IGFBP-3 secretion by PrSC, it conversely reduced the amounts of biologically active IGF-I unbound to IGFBP-3. Immunohistochemical analyses of 49 human prostate cancer tissues showed that IGF I expression in the stroma correlated positively with TGF-beta1 expression in the stroma (r = 0.551, p = 0.0001). Furthermore, TGF-beta1 actually suppressed the growth of DU-145 cells in coculture with PrSC. But, IGFBP-3 proteinases, such as prostate specific antigen (PSA) and matrix metalloproteinase-7 (MMP-7), restored the DU-145 cell growth, suggesting that degradation of IGFBP-3 potentiates cancer growth. Taken together, these results indicated that TGF-beta1 modulates the growth of prostate cancer, either positively or negatively, through the balance between the amounts of IGF-I and IGFBP-3. This complex behavior of TGF-beta1 on the IGF axis is one explanation for the pleiotropic activities of TGF-beta1 on the growth of prostate cancer. PMID- 18507014 TI - Novel link between estrogen receptor alpha and hedgehog pathway in breast cancer. AB - Ligand-dependent constitutive activation of the hedgehog (Hh) pathway is important in the development of various carcinomas including breast cancer. A link between estrogen receptor alpha (ERalpha) and the Hh pathway in human breast cancer is shown here for the first time. In ERalpha-positive cells, estrogen depletion decreased the expression of sonic hedgehog (Shh), a ligand of the Hh pathway, while estrogen supplementation triggered Shh up-regulation. This estrogen-induced Shh expression activated the Hh pathway in a ligand-dependent manner, and increased cell proliferation. These effects were suppressed by ERalpha inhibitors, including ICI 182,780 (ICI), the dominant negative form of ERalpha and small interfering RNA (siRNA) against ERalpha. Consistent with the in vitro data, a positive correlation between ERalpha and Shh expression was found in breast cancer tissues. These data suggest that ERalpha regulates the Hh pathway through Shh induction, and promotes breast cancer development. PMID- 18507015 TI - Prediction of tumour regrowth of pontine glioma using a two-term model. AB - BACKGROUND: Modelling tumour regrowth kinetics and determining tumour burden is critically important in making accurate patient prognoses. PATIENTS AND METHODS: Our two-term model, which describes tumour regrowth after chemotherapy was tested by analyzing tumour sizes in patients with pontine gliomas treated at a major cancer centre during the past 40 years. Tumour measurements at four early time points were used to fit the model, which was then used to predict later tumour sizes. The predictions were compared with observed measurements from clinical charts and this model's performance was compared to that of a quadratic model. RESULTS: The mean of relative errors using the two-term model was 0.186; standard deviation, 0.176. For the quadratic model, the mean of relative errors was 0.412; standard deviation, 0.521. CONCLUSION: The two-term model fits the tumour regrowth data reasonably well and is more accurate than a competing quadratic model used in the same way (p < 0.05). PMID- 18507016 TI - Role of DNA content analysis and immunohistochemistry in the evaluation of the risk of unfavourable outcome in Wilms' tumours. AB - BACKGROUND: Wilms' tumour (WT) is the most common solid tumour affecting young children. Its histological diversity leads to difficulties in predicting the outcome. MATERIALS AND METHODS: Image analysis cytometry and immunohistochemistry with a selected panel of antibodies were performed in 23 cases of WT considered of intermediate risk according to the revised International Society of Pediatric Oncology (SIOP) working classification of renal tumours of childhood. In this series, a tumour was considered aggressive according to its propensity for metastases or its recurrence. RESULTS: Out of the 14 non-aggressive WT, 4 were found to be diploid and 10 were aneuploid including 6 that were heterogeneous for DNA-ploidy. All the tumours presented a low proliferative index and were negative for p53 and p57(kip2) immunostaining. Out of the 9 aggressive tumours, all were aneuploid and 4 were found to be heterogeneous for DNA-ploidy. They all presented a high degree of cell proliferation and 7 were positive for p53 immunostaining. Only two were positive for the p57(kip2) marker. The only fatal case revealed an aneuploid-homogeneous DNA-ploidy analysis, was p53 and p57(kip2) positive and presented a high cell proliferation index. CONCLUSION: A significant correlation between the presence of focal DNA-aneuploidy in Wilms' tumours and adverse prognosis is not established, but some immunohistochemical markers may be useful for the clinical evaluation of these tumours and to help in predicting the risk of an unfavourable outcome. PMID- 18507017 TI - Gene polymorphisms of the NOD-2/CARD-15 gene and the risk of gastric cancer in Germany. AB - BACKGROUND: NOD-2 is involved in the intracellular recognition of bacterial muramyl dipeptides, and three independent polymorphisms of this gene have been identified as risk factors for the development of Crohn's disease. PATIENTS AND METHODS: To study the role of NOD-2 in gastric carcinogenesis, NOD-2 mutations (SNP5: P268S, SNP8: R702W, SNP12: G908R, and SNP13: 3020insC) were genotyped in 171 patients with gastric cancer and 153 controls. RESULTS: Applying a numerical model, SNP5 was found to carry a slightly increased risk (OR = 2.25, 95% CI: 1.05 2.17, p = 0.027) for the development of gastric cancer, whereas SNP8 was similarly distributed between controls and patients with gastric cancer. SNP5 and 8 were found to be genetically linked in both groups (p < 0.02). The allele frequency of SNP12 and 13 were rare and therefore analyzed in subgroups only and not statistically analyzed due to the lack of statistical power. CONCLUSION: NOD 2 is not a risk factor for gastric carcinogenesis in the Caucasian population. PMID- 18507018 TI - Up-regulation of EphB and ephrin-B expression in rhabdomyosarcoma. AB - BACKGROUND: Increased expression of Eph receptors and their ephrin ligands has been implicated in promoting angiogenesis and tumour progression in several malignancies. Here the expression of mRNA for ephrin-B and EphB receptors in rhabdomyosarcoma (RMS) cell lines and primary tumours was investigated. MATERIALS AND METHODS: Expression of mRNA for ephrin-B and EphB receptors in RMS cell lines and primary tumours was measured by real-time RT-PCR and compared with the expression in normal striated muscle. RESULTS: A dysregulation of both ligands and receptors was found in all cell lines. In embryonal tumours, overexpression of ephrin-B1 correlated with overexpression of EphB1 (r = 0.97, p < 0.01) and EphB3 (r = 0.94, p < 0.05); overexpression of ephrin-B2 correlated with overexpression of EphB1 (r = 0.94, p < 0.05), EphB2 (r = 0.88, p < 0.01) and EphB4 (r = 0.76, p < 0.01). In alveolar tumours, no similar correlations were found. A correlation between EphB2 and EphB4 receptors was demonstrated in both tumour types, being positive in embryonal cases (r = 0.81, p < 0.01) and negative in alveolar (r = -1.00, p < 0.01). CONCLUSION: A global up-regulation of ephrin-B and EphB receptors in RMS tumours was found. The correlation between EphB2 and EphB4 receptors suggests a possible role for ephrin-B and EphB receptors in RMS development. PMID- 18507019 TI - Bone marrow-derived endothelial cells contribute to angiogenesis in murine WEHI and JC tumors. AB - The goal of this study was to determine the contribution of bone marrow-derived circulating endothelial progenitor cells to the formation of endothelial cell linings of tumor vessel walls. The proportion of male endothelial cells in female JC and WEHI tumors was measured in male BALB/c mice and in female mice displaying complete marrow chimerism, after receiving male bone marrow cells. The gender origin of the perivascular endothelial cells was determined by fluorescent in situ hybridization (FISH) analysis of adjacent cuts of the tumors, using CD31 and Y-chromosomes as markers. High proportions of male cells were detected in the perivascular endothelial cell linings of the JC (60 +/- 4%) and WEHI (67 +/- 4%) tumors after implantation into normal male mice. Furthermore, in marrow chimeric female mice, very high levels of male cells were observed in the endothelilal cell linings of the tumor vessel walls of both tumor types, after bone marrow transplantation. We conclude that JC and WEHI tumors can serve as a murine experimental model and that bone marrow cells from these can be manipulated and cultured in vitro for use in studies of tumor vessel walls after transplantation into myeloablated recipients. PMID- 18507020 TI - Down-regulation of CD28, TCR-zeta (zeta) and up-regulation of FAS in peripheral cytotoxic T-cells of primary breast cancer patients. AB - BACKGROUND: Several studies have supported the hypothesis that the concept of immuno-surveillance would not be effective in cancer patients. One reason for suppression of antitumor immunity may be attributed to immune impairment of T lymphocytes, which extends beyond the tumor-microenvironment and might effect the peripheral blood. Therefore the aim of this study was to investigate the expression of immunoregulatory antigens in peripheral blood lymphocytes of primary breast cancer patients in comparison with healthy donors. MATERIALS AND METHODS: The peripheral blood immune status of 61 patients with primary breast cancer was analysed by FACS-analysis. The different lymphocytic subpopulations were identified by intracellular/extracellular monoclonal antibodies in three color flow cytometry. The distribution was compared to age-matched healthy female donors (n = 29). RESULTS: The expression of TCR zeta-chain, an important signal complex for T-cell activation and functional integrity of specific immune response, was significantly reduced in the cytotoxic specific T-cell population. Cytotoxic T-cells (CD3+/CD8+) also showed a down-regulation of CD28, the important ligand to the co-stimulatory molecule CD80 (B7.1) on antigen-presenting cells. Moreover, breast cancer patients had significantly more CD95 (FAS) expressing cytotoxic T-cells than their healthy counterparts (p < 0.05). CONCLUSION: The significant up-regulation of CD95 and down-regulation of TCR zeta and CD28 in peripheral cytotoxic T-cells of breast cancer patients leads to the hypothesis of systemic immunosuppression, which could open the door for tumor cell dissemination via the blood stream and which is the subject of ongoing studies. PMID- 18507021 TI - Anticancer mechanism of plumbagin, a natural compound, on non-small cell lung cancer cells. AB - BACKGROUND: Lung cancer is the leading cause of cancer-related deaths in the United States. Prevailing treatment options have limited therapeutic success in lung cancer, particularly non-small cell lung cancer (NSCLC), as it becomes resistant to therapy. Hence, better therapeutic options are immediately required for lung cancer. Plumbagin, a natural compound has been recently examined for its anticancer effect on different cancers. MATERIALS AND METHODS: To determine the anticancer effect of plumbagin on NSCLC cell lines H460 and A549, cell viability, apoptotic, Western blot and reporter assays were performed. RESULTS: Plumbagin significantly inhibited the growth of H460 cells compared to A549 cells, and down regulated the expression of EGFR/Neu and its downstream signaling (Akt, NF kappaB, Bcl-2 and survivin) in H460 cells. In addition, plumbagin up-regulated the expression of p53 and p21(CIP1/WAF1) causing cell cycle arrest in the G2/M phase by down-regulating G2/M regulatory proteins (cyclinB1 and Cdc25B) in H460 cells. Furthermore, it activated the JNK/p38 signaling, leading to caspase-3 activation resulting in the induction of apoptosis. CONCLUSION: Plumbagin exerted anticancer activity on NSCLC cells by modulating the pro-survival and pro apoptotic signaling that causes induction of apoptosis. PMID- 18507022 TI - Effect of depsipeptide on in vitro transfection efficiency of PEI/DNA complexes. AB - The aim of this study was to investigate the effect of depsipeptide on the in vitro transfection efficiency of PEI/DNA complexes. MATERIALS AND METHODS: PEI (polyethylenimine 25K) formed a complex with pcDNA3-CMV-Luc and was investigated for its transfection efficiency in KB human oral carcinoma and Raji human lymphoma cell lines. The transfected cells were then incubated with different concentrations of depsipeptide for 24 h and examined for the transfection efficiency and cell viability. RESULTS: The transfection efficiency in KB cells post-incubated with depsipeptide was not different from the cells transfected with PEI alone. The transfection efficiency in Raji cells post-incubated with 15 nM depsipeptide was significantly increased (p < 0.05) and found to be 76-fold compared to the cells transfected with PEI alone. Depsipeptide had no effect on the cell viability at a concentration of 0.001-1 nM in KB cells and 1-37.5 nM in Raji cells. A high concentration of depsipeptide decreased the cell viability in both cell lines. CONCLUSION: The results demonstrate that PEI/DNA complexes post incubated with depsipeptide can enhance the transfection efficiency in the Raji human lymphoma cell line. The effect of depsipeptide on cell viability was dose dependent. This study suggests that depsipeptide may have potential usefulness for gene delivery in patients with lymphoma. PMID- 18507023 TI - Tumor cell-specific gene expression by 5'-flanking DNA of murine Q5 gene in an adenoviral vector system. AB - BACKGROUND: Regulatory DNA that would induce tumor cell-specific gene expression of arbitrary genes in human cells has been sought. We previously reported that the transcription of mouse Q5 gene was tumor-selective and hypothesized that Q5 5'-flanking DNA had a key role in tumor-selective transcription. MATERIALS AND METHODS: Isolation of 3.9 kb Q5 5'-flanking DNA was carried out and the sequence characteristics determined. Reporter plasmids, recombinant reporter adenoviruses and recombinant reporter lentiviruses were prepared that contained the Q5 5' flanking DNA fragments of various lengths. Expression of these reporter genes was examined in various murine cells in vitro and in vivo and in various human cells in vitro. RESULTS: Adenovirus vectors that had Q5 5'-flanking DNA contained a regulatory region and induced tumor cell-specific gene expression not only in mouse cells but also in human cells. CONCLUSION: Our newly constructed vector system could be utilized in gene therapy of human cancer. PMID- 18507024 TI - A histochemical approach to the evaluation of the in vivo cytotoxicity of the nitrobutadienes (1E,3E)-1,4-bis(1-naphthyl)-2,3-dinitro-1,3-butadiene and methyl (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate in mice liver and kidney. AB - Two new molecules (1E,3E)-1,4-bis(1-naphthyl)-2,3-dinitro-1,3-butadiene (1-Naph DNB) and (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate (1 Naph-NMCB) in previous studies showed interesting antiproliferative activity in vitro. Furthermore, toxicological tests and histological analysis provided promising results, in particular for 1-Naph-NMCB that displayed lower toxic activity both in terms of lethal effect and tissue damage of the main organs. Finally, studies of the antitumour activity in vivo confirmed the efficacy of both molecules, though with some differences in tumour selectivity and levels of activity. In this investigation the activities of some specific enzymes, acid phosphatase (AcPase), alkaline phosphatase (AlkPase), catalase (Cat), succinic dehydrogenase (SDH), glucose-6-phosphatase (G6Pase) and K+ p-nitrophenyl phosphatase (K+ pNPPase) were studied in the liver and kidney as histopathological biomarkers, to assess the effects of the two compounds in organs generally involved in the metabolism and excretion of different drugs. As oxidative stress may also develop as a consequence of the toxic effect of chemicals, reactive oxygen species (ROS) production was evaluated by a histochemical method. The results indicated that some enzyme activities and ROS expression changed in a dose-related manner. Nevertheless, neither in the liver nor in the kidney were dramatic toxic effects evident. By contrast, the variations of some enzyme activities (AlkPase, AcPase, Cat, K+ pNPPase) were interpreted as possible defensive mechanisms for tolerating high dosage of the compounds. PMID- 18507025 TI - Inhibition of PKCalpha activation in human bone and soft tissue sarcoma cells by the selective PKC inhibitor PKC412. AB - BACKGROUND: PKC412, formerly CGP41251, N-benzoylstaurosporine, was initially developed as a selective protein kinase C (PKC) inhibitor, and it preferentially inhibits conventional PKC family members. In this study, the expression of PKCa was examined in human osteosarcoma and MFH cell lines, and the inhibitory effect of PKC412 on the proliferation of the cell lines was evaluated. MATERIALS AND METHODS: Three human osteosarcoma cell lines (KTHOS, MG63 and KHOS) and four human MFH cell lines (TNMY1, GBS-1, Nara-F and Nara-H) were used. The expression of PKCalpha and phosphorylated PKCalpha were analyzed using both Western blotting analysis and immunocytochemical analysis. The effect of PKC412 on cell proliferation was evaluated using the MTS assay technique. RESULTS: PKC412 inhibited cell proliferation of all seven cell lines in a dose- and time dependent manner. Both Western blotting analysis and immunocytochemical analysis revealed that not only PKCalpha but also phosphorylated PKCalpha were expressed in all cell lines incubated with the culture medium without any stimuli. PKC412 suppressed phosphorylation of PKCalpha in all cell lines at a concentration of 1 microM. CONCLUSION: The inhibition of cell proliferation of the human osteosarcoma and MFH cell lines by PKC412 might be due to reduced PKCalpha activity. This suggests PKC412 might be a potent chemotherapeutic agent for human sarcomas. PMID- 18507026 TI - Bee venom induced cell cycle arrest and apoptosis in human cervical epidermoid carcinoma Ca Ski cells. AB - Although it has been previously reported that bee venom (BV) can induce apoptosis in many cancer cell lines, there is no information on the effect of BV on human cervical cancer cells and its molecular mechanisms of action are not fully elucidated. In this study, the possible mechanisms of apoptosis by which BV acts on human cervical cancer Ca Ski cells were investigated. BV induced morphological changes and decreased the percentage of viable Ca Ski cells in a dose- and time dependent manner. Flow cytometric analysis demonstrated that BV induced the production of reactive oxygen species, increased the level of cytoplasmic Ca2+, reduced mitochondrial membrane potential which led to cytochrome c release, and promoted the activation of caspase-3 which then led to apoptosis. BV also induced an increase in the levels of Fas, p53, p21 and Bax, but a decrease in the level of Bcl-2. The activities of both caspase-8 and caspase-9 were enhanced by BV, promoting caspase-3 activation, leading to DNA fragmentation. Based on the DNA fragmentation and DAPI staining, BV-induced apoptosis was mitochondrial-dependent and caspase-dependent. BV also promoted the expression of AIF and Endo G in the Ca Ski cells. Both AIF and Endo G proteins were released from the mitochondria, and then induced apoptosis which was not through activation of caspase. In conclusion, our data demonstrated that BV-induced apoptosis occurs via a Fas receptor pathway involving mitochondrial-dependent pathways and is closely related to the level of cytoplasmic Ca2+ in Ca Ski cells. PMID- 18507027 TI - Culture of primary epithelial adenoma cells from familial adenomatous polyposis patients. AB - BACKGROUND: Colorectal tumors arise from unregulated cell proliferation of the intestinal epithelium through a multistep process the first step usually being premalignant adenomas. Familial adenomatous polyposis patients carry a germ line mutation in the APC gene leading to the development of thousands of polyps, which, if left untreated, lead to cancer. The goal of this study was the establishment of conditions for the culture of epithelial cells composing an adenomatic structure. MATERIALS AND METHODS: All colorectal specimens were obtained from FAP patients within 1-2 hours of surgery. Cells were cultured by standard procedures. Characterization was carried out by immunostaining with pancytokeratin, vimentin and desmoplakin antibodies. RESULTS: A culture protocol that gave rise to epithelial cell growth with high efficiency and efficacy was established. Successful subculturing of the cell sheets took place only when dispase prepared in Ca2+ and Mg2+ free medium, was used to digest polyp tissue taken from FAP patients. By using immunostaining these cells were characterized as epithelial. CONCLUSION: The protocol we developed here provides a means of preparing cell cultures from human colorectal adenomas, which aid in the research of the transition from adenoma to carcinoma. PMID- 18507028 TI - Prognostic value of protease-activated receptor-1 (PAR-1) and matrix metalloproteinase-1 (MMP-1) in gastric cancer. AB - BACKGROUND: Protease-activated receptors (PARs) are proposed to be involved in the invasive and metastatic processes of various types of cancer. Matrix metalloproteinase-1 (MMP-1) plays a role in cancer invasion and tissue remodelling. It has been reported that MMP-1 can alter the behavior of cancer cells through PAR-1 to promote cell migration and invasion. We considered whether the expression of PAR-1 and MMP-1 has relevance to progression in gastric cancer. MATERIALS AND METHODS: An immunohistochemical study was carried out on 129 samples of gastric cancer using anti-PAR-1 and anti-MMP-1 mouse monoclonal antibodies. Associations between immunostaining and clinicopathological factors were analyzed statistically. RESULTS: There were 58 carcinomas positive for PAR-1 expression. The expression of PAR-1 was associated with the depth of wall invasion and peritoneal dissemination. There were 42 carcinomas positive for both PAR-1 and MMP-1 expression which was associated with the histological stage, depth of wall invasion, lymph node metastasis and peritoneal dissemination. These patients had a significantly poorer prognosis than those with expression-negative tumors. Multivariate analysis indicated that PAR-1 expression and combined PAR-1 and MMP-1 expression were independent prognostic factors. CONCLUSION: The results led us to believe that the expression of PAR-1 and MMP-1 is associated with the progression of gastric cancer and an independent prognostic predictor. PMID- 18507029 TI - Valproic acid sensitizes K562 erythroleukemia cells to TRAIL/Apo2L-induced apoptosis. AB - BACKGROUND: Selectively targeting death receptors to trigger apoptosis in cancer cells appears ideal in cancer therapy. The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) is of great interest since it has been shown to predominantly kill cancer cells without toxic effects on normal counterparts, thus representing a promising anticancer agent. However, resistance towards TRAIL/Apo2L treatment has also been described. To overcome this obstacle, co-administration of TRAIL/Apo2L plus several compounds, including histone deacetylase inhibitors (HDACi), has been attempted as a strategy to restore cancer cell sensitivity to TRAIL-induced apoptosis. In recent years, the clinical application of HDACi has been largely explored for their ability to modulate gene transcription, block cell division cycle, inhibit cell proliferation, induce cellular differentiation and apoptosis. MATERIALS AND METHODS: The ability of valproic acid (VPA), a well-known HDACi, to sensitise the K562 cell line, derived from a human leukemia, to TRAIL/Apo2L-mediated apoptosis was evaluated. VPA was selected since it is currently used in clinical practice and its pharmacokinetic, pharmacodynamic and bioavailability are known. RESULTS: When applied with TRAIL/Apo2L, VPA increased cell death and caspase-3 activity by 4-fold compared to the treatment with TRAIL/Apo2L alone. VPA sensitized K562 cells to TRAIL/Apo2L mediated apoptosis by increasing the expression of DR4 and DR5 by 3- and 14-fold respectively. In addition, VPA per se, in the absence of TRAIL/Apo2L, reduced the expression of antiapoptotic factors, such as c-FLPs, associated with DISC, and Bcl-2/Bcl-X(L), associated with mitochondria, acting on both extrinsic and intrinsic apoptotic pathways. CONCLUSION: Our results demonstrated the ability of VPA to sensitize TRAIL/Apo2L-resistant cells to apoptosis, thus providing an attractive approach for the treatment of leukemias and other proliferative malignancies. PMID- 18507030 TI - Imup-1 and imup-2 overexpression in endometrial carcinoma in Korean and Japanese populations. AB - BACKGROUND: Endometrial carcinoma is a well known complex gynecological disorder. Our team suggests that this tumor is related to immortalization-up-regulated protein 1 and 2 (imup-1, imup-2), which are known to be involved in SV40-mediated immortalization. PATIENTS AND METHODS: RT-PCR and immunohistochemical staining were used to examine the mRNA expression levels and intracellular localization of imup-1 and imup-2 in endometrial carcinomas from Korean and Japanese patients. RESULTS: The imup-1 (4.1- and 23.6-fold) and imup-2 (4.8- and 2.7-fold) mRNA expression levels in endometrial carcinomas from both Korean and Japanese women were significantly higher than in normal endometrial tissues (p < 0.01). Strong expression of the IMUP-1 and IMUP-2 proteins were found in the tumor cytoplasm as well as in the nuclei. CONCLUSION: These findings demonstrate the up-regulation of imup-1 and imup-2 in human endometrial carcinomas and indicate that these molecules play a role in endometrial carcinogenesis in both Korean and Japanese patients. PMID- 18507031 TI - Prognostic value of eicosanoid pathways in extrahepatic cholangiocarcinoma. AB - BACKGROUND: Chronic inflammation of the bile duct is linked to an increased risk for the development of cholangiocarcinoma. Arachidonic acid and linoleic acid oxidation through cyclooxygenase and lipoxygenase--two major pro-inflammatory pathways--have rarely been investigated in extrahepatic cholangiocarcinoma. MATERIALS AND METHODS: Paraffin-embedded specimens from 51 resected adenocarcinomas of the extrahepatic bile duct were immunostained for cyclooxygenase 2 (COX-2) and 5-lipoxygenase (5-LOX) to evaluate their intracellular distribution and prognostic value. RESULTS: Cholangiocarcinoma had significantly higher levels of 5-LOX and COX-2 expression compared with normal tissue (p = 0.015). High expression of nucleus-located 5-LOX was significantly associated with intensive staining for COX-2, (p = 0.023). Median disease-free survival (DFS) in patients with low expression of 5-LOX was significantly better than in patients with high expression of 5-LOX (log rank p = 0.046). DFS in patients with low COX-2 expression was also significantly better than DFS in patients with high COX-2 expression (log rank p = 0.0187). CONCLUSION: The present study demonstrates that 5-LOX and COX-2 protein expression was increased in cholangiocarcinoma suggesting that these two enzymes might be of prognostic value and offer a potential additional adjuvant therapeutic approach to this disease. PMID- 18507032 TI - Radiofrequency ablation of experimental bone metastases in nude rats. AB - PURPOSE: Animal models are indispensable to investigate bone metastasis and to test different preclinical therapy options. Radiofrequency ablation is an upcoming technique for palliating pain from bone metastases. The aim of this study was to generate osteolytic lesions and to enable a technique to achieve access to the bone to successfully carry out radiofrequency ablation. MATERIALS AND METHODS: Human breast cancer cell line MDA-MB-231 (10(5) tumor cells) was implanted into the femur of 10 nude rats using a drill hole after arthrotomy of the knee joint and opening of the femur through the notch. Weekly CT- and MRI scans were performed to document number and size of bone metastases. Radiofrequency ablation (22G bipolar and impedance-controlled RF-applicator, 2-4 Watt, 3 min application time) was carried out. One week after RFA, the animals were sacrificed and macroscopic and histological examination followed. For statistical analysis, paired comparison procedures were used. RESULTS: Inoculation of the tumor cells was well tolerated. The mean time of the surgical procedure was 6 minutes. All animals developped local bone metastases. Mean time to metastasis was 8 weeks (range 7-10 weeks) after tumor cell implantation. No leakage of tumor cells and no soft part metastases occurred. Radiofrequency ablation was performed without complications. Imaging showed a complete ablation of the bone tumor in all rats. Histological findings confirmed a circular necrosis with an extensive destruction of tumor cells leaving a necrosis cavity. CONCLUSION: The experimental model presented here describes the first time the ability to carry out radiofrequency ablation in nude rats with intrafemoral induced osteolytic metastases of human breast cancer. RFA in human breast cancer cell line in nude rats is a feasible and useful possibility to evaluate and to test different RF-procedures. Additional treatment options like local chemotherapy or chemoembolization can be performed. PMID- 18507033 TI - Sensitivity of Hodgkin's lymphoma cell lines to the cell cycle inhibitor roscovitine. AB - BACKGROUND: The prognosis of patients with Hodgkin's lymphoma (HL) has improved in recent decades. However, not all patients can be cured and the development of alternative treatment strategies is necessary. MATERIALS AND METHODS: Gene expression in HL cell lines was analyzed using DNA microarrays and both conventional and quantitative reverse transcriptase-polymerase chain reaction. Sensitivity of HL cell lines to the cell cycle inhibitor roscovitine was assessed in vitro. RESULTS: All HL cell lines express high levels of cyclin D2. Treatment of HL cells with roscovitine induced cell death in some cell lines whereas other cell lines were resistant to roscovitine. Roscovitine-sensitive cell lines were characterized by expression of T-cell markers and expressed high levels of the unusual cytokine interleukin-26. CONCLUSION: Roscovitine is a cytotoxic drug for a subpopulation of HL cells and might be an interesting agent for the treatment of patients with HL. PMID- 18507034 TI - Rosiglitazone induces caveolin-1 by PPARgamma-dependent and PPRE-independent mechanisms: the role of EGF receptor signaling and its effect on cancer cell drug resistance. AB - BACKGROUND: Caveolin-1, a key component of plasma membrane caveolae, has been implicated in the regulation of cancer cell growth and survival. Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a ligand-activated nuclear receptor which plays a pivotal role in many cellular processes. Activation of PPARgamma by its ligand rosiglitazone upregulates caveolin-1 mRNA and protein in human carcinoma cells. MATERIALS AND METHODS: We have used specific signaling inhibitors to dissect the mechanisms of caveolin-1 mRNA and protein induction by rosiglitazone, determined by RT-PCR and Western blotting, respectively. ROS generation was measured by flow cytometry and cell survival was determined by the MTT assay. RESULTS: We show that in HT-29 human colon cancer cells the induction ofcaveolin-1 by rosiglitazone is inhibited by the EGF receptor (EGFR) blocker AG1478. Moreover, rosiglitazone stimulates EGFR phosphorylation, while direct activation of EGFR by EGF up-regulates caveolin-1 mRNA. Inhibitors of Src and the Mek1-Erk1/2 and p38 MAP kinase pathways also inhibit up-regulation of caveolin-1 by rosiglitazone. Furthermore, rosiglitazone stimulates formation of superoxide anions, whereas induction of caveolin-1 expression by rosiglitazone is attenuated by the antioxidant N-acetyl-cysteine. Finally, rosiglitazone increases the resistance of HT-29 cells to doxorubicin and to hydrogen peroxide. The caveolin-1 gene promoter lacks a canonical PPARgamma response element (PPRE) and a PPRE reporter construct is not sensitive to EGF or EGFR inhibition. CONCLUSION: Our findings indicate that up-regulation of caveolin-1 by rosiglitazone requires superoxide formation and the activation of Src, EGFR, and the Mek1-Erk1/2 and p38 MAP kinase pathways. We suggest a novel mode of action of PPARgamma ligands in the regulation of caveolin-1, and possibly other genes devoid of a PPRE in their promoters, which involves the coordinate activation of PPARgamma and intracellular signaling pathways. PMID- 18507035 TI - miR-21 microRNA expression in human gastric carcinomas and its clinical association. AB - BACKGROUND: MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs that have been shown to be aberrantly expressed in many human carcinomas. Of these miRNAs, miR-21 appears to be important in tumorigenesis given its up regulation in almost all types of human cancer examined. However, its association with the clinicopathological features of human gastric cancer has yet to be addressed. PATIENTS AND METHODS: Cancer tissues and corresponding normal tissues from 37 patients with gastric cancer were examined for the expression level of miR-21 using quantitative PCR and the clinical relevance of miR-21 was statistically analyzed. RESULTS: miR-21 was overexpressed in 92% (34/37) of the gastric cancer samples examined. However, the patients with higher miR-21 expression did not have a worse prognosis. CONCLUSION: miR-21 could serve as an efficient diagnostic marker for gastric cancer, but does not affect the clinical prognosis of gastric cancer patients. PMID- 18507036 TI - Analysis of EGFR overexpression, EGFR gene amplification and the EGFRvIII mutation in Portuguese high-grade gliomas. AB - BACKGROUND: Patients with malignant gliomas do not respond to any current therapy. Epidermal growth factor receptor (EGFR) controls several oncogenic processes, being frequently up-regulated in gliomas due to overexpression, gene amplification and gene mutation. EGFR inhibitors are being tried in gliomas, yet the molecular determinants of therapeutic response are unclear. MATERIALS AND METHODS: EGFR overexpression, EGFRvIII mutation and EGFR amplification were determined by immunohistochemistry and chromogenic in situ hybridization (CISH) in 27 primary glioblastomas (GBM), 24 anaplastic oligodendrogliomas (AO) and four anaplastic oligoastrocytomas (AOA). RESULTS: EGFR overexpression was associated with EGFR amplification, being found in 48% and 53% GBM, 33% and 40% AO and 75% and 67% AOA, respectively. EGFRvIII was found in 22% GBM, 8% AO and was absent in AOA. No association was observed between EGFR alterations and patient survival. CONCLUSION: We characterized, for the first time, EGFR molecular alterations in Portuguese patients with malignant glioma and identified a subpopulation of patients presenting putative biomarkers for EGFR-based therapies. PMID- 18507037 TI - Positive correlation of cell-free DNA in plasma/serum in patients with malignant and benign breast disease. AB - BACKGROUND: Circulating cell-free (ccf) DNA is measurable in healthy individuals and in higher concentration in patients with benign and malignant breast disease (BD). PATIENTS AND METHODS: In paired plasma and serum samples ccf DNA was extracted and quantified by real-time quantitative PCR for the glyceraldehyde-3 phosphate dehydrogenase (GAPDH) gene. RESULTS: The concentration of ccf DNA in serum was higher in patients with benign and malignant BD (p = 0.023/p = 0.001) compared to healthy controls, whereas ccf DNA in plasma was higher in patients with malignant BD compared to patients with benign BD or healthy controls (p = 0.012/0.007). The ccf DNA correlated significantly between plasma and serum samples in patients with benign (p = 0.01; R: 0.677) as well as malignant BD (p = 0.01; R:0.713). CONCLUSION: The positive correlation between ccf DNA in plasma and serum in patients with benign as well as malignant BD, might have a diagnostic value for discriminating between malignant and benign BD. PMID- 18507038 TI - Anticancer activity of 3-demethylubiquinone Q2. In vivo experiments and probable mechanism of action. AB - BACKGROUND: 3-Demethylubiquinone Q2 (3DMUbQ2), isolated from the ascidian Aplidium glabrum and later synthesized, is known as a natural product inhibiting EGF-induced malignant JB6 P+ Cl 41 cell transformation. However, its in vivo anticancer properties and probable mechanism of this action have not been studied. MATERIALS AND METHODS: Preventive and curable effects of3DMUbQ2 on mice with inoculated Ehrlich carcinoma tumors were examined by magnetic resonance tomography. Capability to inhibit human tumor cell colony growth and induce their apoptosis was investigated using the anchorage-independent phenotype expression assay in soft agar and flow cytometry. RESULTS: 3DMUbQ2 inhibits the growth of the solid Ehrlich carcinoma in mice, especially using the prophylactic scheme of administration (50% inhibition). It inhibits the phenotype expression of HT-460, HCT-116 and SK-MEL-28 human tumor cells and induces apoptosis of these cell lines, as well as that of HL-60 and THP-1 tumor cells. CONCLUSION: 3DMUbQ2 and other related marine polyprenylquinones have potential for development of a new antitumor agent in cancer prophylactics and treatment and should be further investigated. PMID- 18507039 TI - Antitumoral and antiangiogenic efficacy of bisphosphonates in vitro and in a murine RENCA model. AB - BACKGROUND: Bisphosphonates have shown direct antitumoral activity in vitro, in vivo and even in clinical studies, but the exact mechanism for this has not yet been elucidated. In this study the antiangiogenic potency of zoledronic acid and clodronate were evaluated. MATERIALS AND METHODS: The effects of zoledronic acid and clodronate on the proliferation of endothelial cells and different tumor cells, and on the activity of protein kinases were investigated. Furthermore in vitro experiments were performed to evaluate the underlying antiangiogenic mechanism of action. Both bisphosphonates were examined in vivo at different doses and in daily subcutaneous application in a murine renal cell carcinoma model (RENCA). The antiangiogenic activity was evaluated by immunohistochemical staining (CD31) and by determination of mouse vascular endothelial growth factor (VEGF) serum concentration. RESULTS: Zoledronic acid and clodronate inhibited proliferation of endothelial cells at lower concentrations than the different tumor cell lines did. This effect was more pronounced for zoledronic acid. The activity of almost all tested kinases was inhibited by zoledronic acid, whereas clodronate showed no effect. In the RENCA model, a significant effect of zoledronic acid on the primary tumor in a bell-shaped dose response curve with the highest efficacy between 100 Bg/kg 2xd and 200 Bg/kg 1xd, was observed. The mean vessel density (MVD) was significantly reduced by both bisphosphonates at different concentrations. This is the first report on increased mouse VEGF serum concentrations in the RENCA model. CONCLUSION: The results indicate that these bisphosphonates, particularly zoledronic acid, possess antitumoral and antiangiogenic activity. PMID- 18507040 TI - A facile microfluidic method for production of liposomes. AB - BACKGROUND: Ethanol injection is widely used in liposome preparation. However, the parameters determining particle size distribution of the liposomal preparation has not been fully defined. MATERIALS AND METHODS: A syringe pump driven microfluidic injection device was used to produce liposomes under different conditions. RESULTS: Particle size of the liposomes was decreased with decrease in needle diameter (or increase in hydrodynamic pressure), decrease in lipid concentration in the alcohol solution, decrease in phase transition temperature (T(m)) of the lipid bilayer and the absence of cholesterol (or decrease in, membrane rigidity). CONCLUSION: The device used is simple to adopt and can be used for affordable production of liposomes with tunable particle size. PMID- 18507041 TI - Predicting resistance to platinum-containing chemotherapy with the ATP tumor chemosensitivity assay in primary ovarian cancer. AB - BACKGROUND: Most patients with primary epithelial ovarian cancer (PEOC) treated with carboplatin/paclitaxel will relapse between one to two years. Our purpose was to define the optimal calculation method for the adenosine triphosphate-tumor chemosensitivity assay (ATP-TCA) applied to PEOC treated with platinum-containing chemotherapy and to analyze its predictive relevance. MATERIALS AND METHODS: ATP TCA results from 80 PEOC specimens were analyzed applying three different methods: 50% inhibition concentration, sensitivity index (IndexSUM), and area under curve by testing multiple cut-off levels. Correlation between in vitro results and clinical outcome was performed for 61 (76%) patients by univariative and multivariative analysis. Tumor recurrence 6 months after chemotherapy was classified as platinum-resistance. RESULTS: The IndexSUM set at > 250 had the highest test sensitivity, specificity, positive and negative predictive value of 90%, 43%, 62% and 81%, respectively. Patients whose tumors were shown to be resistant by ATP-TCA had a higher risk for recurrence (RR) compared to those who tested as sensitive (p < 0.003, RR = 3.3, 95% CI = 1.2-9.4). This result was confirmed after adjustment for FIGO stage by logistic regression (p < 0.004, Odds ratio = 8.3, 95% CI = 1.9-35.5). In multivariate analysis ATP-TCA and the FIGO stage were independent predictive factors of early recurrence. CONCLUSION: ATP TCA results in combination with the use of IndexSUM > 250 are best able to predict platinum resistance. PMID- 18507042 TI - Characterization of Akt overexpression in MiaPaCa-2 cells: prohibitin is an Akt substrate both in vitro and in cells. AB - Akt (PKB) is a serine/threonine protein kinase that plays an important role in the transduction of signals affecting apoptosis, cell proliferation and survival. The Akt gene is frequently hyperactivated in tumors and has been shown to be amplified in a number of types of human cancers. Furthermore, Akt activity is elevated in cell lines with the mutated PTEN tumor suppressor gene. These studies establish Akt as an attractive target for cancer therapy. To determine the roles of Akt1, Akt2 and Akt3 in signal transduction, constitutively active Akt1, Akt2 and Akt3 was ectopically overexpressed in human pancreatic MiaPaCa-2 cells. The three Akt stable clones were characterized to determine their effects on transformation and proliferation. Compared to a vector control, the three Akt clones were able to drive cellular proliferation even in reduced serum conditions. Furthermore, in soft-agar assays, the Akt clones showed an 25-38% increase in colony formation in 2% serum. Our results indicate that all three forms of Akt may have protective effects within the cell depending on the type of apoptotic stimuli. Using 2D-PAGE comparisons between parental and Akt overexpressing cells, we attempted to determine novel targets of Akt phosphorylation. In this study, we identified prohibitin as a substrate for Akt both in vitro and in vivo. These studies suggest that Akt may regulate the cellular function of prohibitin via its phosphorylation. PMID- 18507043 TI - Anticancer effects of Annona glabra plant extracts in human leukemia cell lines. AB - Annona glabra (pond apple), a tropical tree growing wild in the Americas and Asia, is used in traditional medicine against several human ailments, including cancer. To validate the ethnopharmacological claims against cancer, the anticancer effects of alcoholic extracts prepared from pond apple leaves, pulp and seed, were investigated in human leukemia cell lines. The alcoholic extracts were not cytotoxic to normal human lymphocytes. However, extracts were highly cytotoxic to drug sensitive (CEM) and multidrug-resistant leukemia (CEM/VLB) cell lines. The seed extract was more potent than leaf and pulp extracts, and the cytotoxicity values were significantly lower than that for adriamycin. The seed extract caused a concentration-dependent increase in the percentage of the sub G0/G1, as well as G0/G1 cell population, contributing to the cytotoxicity. The sub G0/G1 population increased from 2.2 to 7.0% in CEM and from 1.0 to 10.7% in CEM/VLB cell lines, when the cells were treated with 0-10 Bg/ml seed extract. Treatment of CEM and CEM/VLB cells with seed extract induced apoptosis and necrosis in both sensitive and resistant leukemia cells in a concentration dependent manner. The seed extract at 2 and 5 Bg/ml enhanced cellular daunorubicin accumulation, indicating the competitive P-glycoprotein binding ability and drug-resistance reversal effect. Treatment of CEM and CEM/VLB cells with seed extracts also up-regulated the expression of cyclin kinase inhibitor (WAF1/p21) contributing to the arrest of cells at the G0/G1 phase of the cell cycle. These results support the traditional use of A. glabra and the alcoholic seed extract is a potent source of anticancer compounds that could be utilized pharmaceutically. PMID- 18507044 TI - HER3 expression in cutaneous tumors. AB - BACKGROUND: In order to elucidate the role of the receptor tyrosine kinase HER3, the expression characteristics in different tissues of cutaneous malignancies and in normal skin were compared. MATERIALS AND METHODS: In this study HER3 expression was evaluated by RT-PCR analysis and immunohistochemistry from different tissue specimens of cutaneous tumors like nevi, primary malignant melanomas, basal cell carcinoma, squamous cell carcinoma and malignant melanoma metastases and normal skin samples and graded into weak, moderate and strong expression. Associations of tumor thickness in these specimens with HER3 expressions were also analyzed. RESULTS: HER3 expression was found in 63% (10/16) of the basal cell carcinomas, in 4/5 of squamous cell carcinomas and in one Merkel cell carcinoma. Within the group of different malignant melanomas, HER3 expression was detected in 35% of the nodular malignant melanomas (6/17) and in 9/19 of the superficial spreading melanomas, including 2 lentigo malignant melanomas. The majority of melanomas with a higher tumor thickness expressed HER3, and 85% of melanoma metastasis were HER3-positive. CONCLUSION: HER3 expression was associated with hyperproliferate tumor stages and suggested that HER3 expression could reflect an increased malignant potential in cutaneous lesions. PMID- 18507045 TI - 18F-FDG small animal PET for early detection of human anaplastic large cells lymphoma xenograft in immunocompromised mice. AB - The purpose of the present study was to assess if small animal PET is useful for serially monitoring the development of a human anaplastic large cell lymphoma (ALCL) murine xenograft and for the early selection of tumour bearing animals. The human ALCL Karpas 299 cell line was subcutaneously injected in 6-week-old NOD/SCID (non-obese diabetic/NCrCrl- Prkdc) mice (10(7) cells/mouce in 150 pil FBS) at the right flank level. Small animal 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET) was serially performed (intravenous injected dose: 20 MBq in < 0.15 ml, uptake time: 60 min, image acquisition: 1 bed position of 15 min): early PET at 2 days after cell inoculation in 4/8 mice and at 4 days in the remainig 4/8, later PET scans were performed in all the animals at 7, 14, 21 and 28 days after inoculation. The images were evaluated visually and the tumour to background ratio (TBR) was used for semiquantitative analysis. Pathology sections were obtained in all cases. PET detected the presence of the tumour as early as seven days after inoculation in 4/8 mice and at 14 days in 2/8. Of the two remaining mice, one died after the first PET scan (thus preventing any evaluation of detection time) while the other showed a microscopic neoplastic infiltration at tracheal level at autopsy. Mean TBR progressively increased in all positive cases, particularly in the first 3 weeks, reaching a plateau afterwards. PET was positive in 6/8 (75%) animals, detecting the presence of viable tumour cells earlier than macroscopic evaluation, thus may be used for the early identification of tumour bearing animals. PMID- 18507046 TI - Loss of heterozygosity, microsatellite instability and TP53 gene status in ovarian carcinomas. AB - BACKGROUND: Microsatellite instability (MSI) and loss of heterozygosity (LOH) are frequent events in ovarian carcinogenesis; however, little is known as to their clinical significance and association with other molecular lesions. MATERIALS AND METHODS: Twelve microsatellite markers for MSI and LOH analysis were used in 64 ovarian carcinomas with known TP53 mutational status. The clinical importance of molecular alterations was evaluated in a uniform subgroup of patients treated with platinum-based regimens. RESULTS: LOH was detected in order of frequency at 17p13.3 (D17S926, 79%), 17p13.1 (TP53 locus, 69%), 13q14 (RB, 60%), 3p21 (D3S1611, 32.5%), 8q21 (D8S1811, 22%), 11p14/13 (D11S904, 19%), 10qter (D10S197, 13%) and 2p16-21 (D2S123, 11%). LOH at the RB1 locus showed association with LOH at the TP53 locus (p = 0.01). Platinum sensitivity was associated with heterozygosity at the TP53 locus (p = 0.006). Only one tumor displayed microsatellite instability in one marker (RB) only. CONCLUSION: Our results suggest that LOH at the 17p D17S926 locus in ovarian cancer is an earlier molecular event than LOH at the TP53 locus. Inactivation of TP53 and RB1 genes may have a synergistic effect in ovarian tumorigenesis. PMID- 18507047 TI - Quantitative structure-cytotoxicity relationship analysis of 5 trifluoromethyloxazole derivatives by a semiempirical molecular-orbital method with the concept of absolute hardness. AB - The most stable conformation of twelve 5-trifluoromethyloxazole derivatives was calculated by CONFLEX 5. The optimized structure was determined by CAChe Worksystem 4.9 PM3 method, in the presence (COSMO) or absence (non-COSMO) of water. Higher correlation coefficients for all descriptors were found under COSMO, as compared with non-COSMO conditions. Good correlation was found between the cytotoxicity of these compounds and the electron affinity, ionization potential, highest occupied molecular orbital energy (E(HOMO)), lowest unoccupied molecular orbital energy (E(LUMO)), absolute hardness (eta) and reactivity index (omega). On the other hand, there was generally no clear-cut correlation between CC50 and the heat of formation, stability of hydration, dipole moment, absolute electron negativity (chi), molecular weight, maximum length of molecule, with some exceptions. The cytotoxic activity of 5-trifluoromethyloxazole derivatives became maximum at log p = 4.6. The concept of absolute hardness is applicable in estimating the cytotoxicity of 5-trifluoromethyloxazoles, using an eta-chi activity diagram. PMID- 18507048 TI - Biphenylalkylacetylhydroquinone ethers suppress the proliferation of murine B16 melanoma cells. AB - Hydroquinone, an activator of caspase-9 activity via reactive oxygen species, and farnesol, a post-translational down-regulator of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity suppress the growth of murine 816 melanoma cells. Our previous studies have shown that farnesyl-O-acetylhydroquinone has a markedly greater growth-suppressive activity than that predicted by the responses to the parent compounds. Perillyl alcohol, a modulator of small G-protein activity, and biphenyl compounds, activators of Fas-mediated death pathways, suppress B16 growth. A similar synergistic increase in the potency of each compound when ether linked to acetylhydroquinone is reported. Perillyl-O-acetylhydroquinone, biphenylethyl-O-acetylhydroquinone and biphenylpropyl-O-acetylhydroquinone had dose-dependent impacts on the proliferation of B16 cells with 50% inhibitory concentration (IC50) values of 8.0, 4.2 and 1.4 micromol/L, respectively. The growth-suppression effected by biphenylpropyl-O-acetylhydroquinone was accompanied by a dose-dependent arrest at the G1/S interface of the cell cycle, an impact greater than that previously reported for farnesyl-O-acetylhydroquinone (IC50 = 2.5 micromol/L). These new hydroquinone derivatives may have potential in cancer chemoprevention and/or therapy. PMID- 18507049 TI - Hypoxia increases adhesion and spreading of MG-63 three-dimensional tumor spheroids. AB - The effects of hypoxia on adhesion and spreading of MG-63 human osteosarcoma spheroids were investigated. Hypoxia was induced in 2-day-old, small spheroids and verified by HIF-1alpha expression. Changes in adhesion were examined on both tissue culture plates and plates coated with fibronectin or collagen while spreading was analyzed in cocultures of MG-63 spheroids seeded on primary fibroblasts grown as a monolayer. In order to better distinguish the two different cell types, MG-63 cells were previously stably transfected with the green fluorescent protein EGFP-vector. Changes in the expression of molecules involved in tumor adhesion and spreading, such as two key integrins (fibronectin receptor, alpha5, and collagen receptor, alpha2) and fibronectin were also examined. The results indicate that hypoxia increases adhesion of spheroids and enhances their ability to spread into the surrounding fibroblast cell culture. These changes in adhesion and spreading are accompanied by concomitant variations in the expression of alpha5 and alpha2 integrins and fibronectin. PMID- 18507050 TI - Lack of association among polymorphic xenobiotic-metabolizing enzyme genotypes and the occurrence and progression of oral carcinoma in a Brazilian population. AB - BACKGROUND: A case control association study was carried out to investigate polymorphisms in genes CYP1A1 (3801T > C), GSTM1, and GSTT1 (null genotypes) and oral squamous cell carcinoma (OSCC), including a correlation with some histopathological findings (tumor size, lymph node invasion and degree of tumor differentiation). PATIENTS AND METHODS: The patients (n = 91) and the controls (n = 81) were matched by age, sex, ethnicity and smoking habits. The molecular analysis was carried out using Polymerase Chain Reaction-Restrict Length Polymorphisms PCR-RFLP (CYP1A1) and Multiplex-PCR (GSTM1/GSTT1). RESULTS: No association was found for any of the studied genes: CYP1A1 (odds ratio (OR) = 1.24; 95% Confidence Interval (CI) = 0.67-2.31), GSTM1 (OR = 0.61; CI 95% = 0.33 1.11), and GSTT1 (OR = 1.24; CI 95% = 0.65-2.38). The analysis of combining genotypes also showed lack of association. Comparison with the histopathological findings did not, in general, detect any statistically significant differences. CONCLUSION: CYP1A1, GSTM1 and GSTT1 polymorphisms do not appear to influence the genetic susceptibility to OSCC or the progression to more advanced stages. PMID- 18507051 TI - A prenylation inhibitor (sodium phenylacetate) differently affects MCF-7 cell death when ras is overexpressed, partly involving P42/44, JNK and P38 kinase activations. AB - BACKGROUND: Sodium phenylacetate (NaPa) inhibits breast cancer cell proliferation decreasing prenylation of small G proteins including Ras. MATERIALS AND METHODS: Aponecrosis induced by NaPa in MCF-7 and MCF-7ras breast cancer cells was evaluated by measuring Annexin V/PI labelling by flow cytometry. Specific inhibitors of p42/44 (PD 98059), p38 (SB 600125) and JNK (SP 202190) in association with NaPa were also tested. Mitogen-activated kinase (MAPK) activation was measured by immunoprecipitation. RESULTS: NaPa induced cell death more efficiently (80%) in the MCF-7ras cells compared to the MCF-7 cells (60%). NaPa activated ERK 1/2 and its combination with PD 98059 decreased cell death in the MCF-7ras cells in contrast to the MCF-7 cells. Combination of NaPa with specific inhibitors of both JNK and p38 kinases also partly decreased MCF-7ras cell death. CONCLUSION: NaPa induced cell death differently when ras was overexpressed in breast cancer cells, partly involving p42/44, JNK and p38 pathways. PMID- 18507052 TI - P53Arg72 homozygosity and its increased incidence in left-sided sporadic colorectal adenocarcinomas, in a Greek-Caucasian population. AB - BACKGROUND: The current case-control study was conducted in order to elucidate any possible association of the single nucleotide polymorphism (SNP) of codon 72 of the p53 gene (Arg72Pro) and sporadic colorectal adenocarcinoma development in a Caucasian population in Greece. The distribution of its alleles, in relation to many clinical parameters of the cancer group, was also investigated. MATERIALS AND METHODS: Genomic DNA samples from 93 sporadic colorectal adenocarcinoma cases and 95 healthy controls (age and ethnicity matched) were used to genotype the p53 codon 72 polymorphism. RESULTS: A strong association of the homozygous 72Arg allele with the development of colorectal cancer was observed (Chi-Square = 11,212, p = 0.001, O.R = 2.902, 95% (CI) = 1.540-5.469, for Arg/Arg vs. Arg/Pro and Pro/Pro). When tumor location was accounted for, the Arg/Arg carrier genotypes were associated with an increased incidence of left colon cancer (Chi Square = 5.256, p = 0.026, OR = 2.975, 95% (CI) = 1.150-7.699). CONCLUSION: p53Arg homozygosity is associated with the development of sporadic colorectal adenocarcinoma, in the Greek-Caucasian population studied and this polymorphism may have a significant prognostic value, where tumor location is concerned. PMID- 18507053 TI - Crude extracts of Solanum lyratum induced cytotoxicity and apoptosis in a human colon adenocarcinoma cell line (colo 205). AB - The effects of the crude extract of Solanum lyratum (SLE) on human colon cancer colo 205 cells were investigated. The cell viability, morphological changes of the cells, cell cycle arrest, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (deltapsi(m)) and cell cycle- and apoptosis associated protein levels and gene expressions were examined in colo 205 cells after exposure to various concentrations of SLE for different time periods. The results indicated that SLE decreased the percentage of viable colo 205 cells accompanied by morphological changes. The most effective concentration of SLE was 300 pg/ml (SLE 300) and this concentration was used for further investigations. SLE induced S-phase arrest and apoptosis (sub-G1) in the colo 205 cells and those effects were dose- and time-dependent. DAPI staining and DNA gel electrophoresis confirmed that SLE induced apoptosis in colo 205 cells. Flow cytometric analysis also showed that SLE 300 promoted ROS production and decreased the deltapsi(m). Western blotting analysis indicated that SLE 300 increased Bax levels and decreased Bcl-2 levels, which caused the loss of deltapsi(m) followed by cytochrome c release and caspase-9 and -3 activation, finally leading to apoptosis. SLE 300 also promoted p53 and p27, but decreased the levels of cyclin B1 thus causing S-phase arrest. The gene expression associated with those proteins was also confirmed by PCR methods. The findings show that SLE might be used as a colon cancer therapeutic agent in the future. PMID- 18507054 TI - Cell proliferation and migration in glioblastoma multiforme cell lines are influenced by insulin-like growth factor I in vitro. AB - BACKGROUND: Malignant gliomas continue to be a therapeutic challenge. One of the major problems is the early and rapidly infiltrating tumor growth. The role of the insulinlike growth factor (IGF) system in the progression of malignant glioma growth is poorly understood. In this study we investigated the expression of different members of the IGF system in malignant glioma cells and the influence of IGF-I and -II on the proliferation and migration of human glioma cell lines in vitro. MATERIALS AND METHODS: Expression of IGF-I and -II, IGF-receptor I and II and IGF binding proteins (IGFBP) 1 to 6 was analysed by PCR in cell lines T98G, A172, 86HG39 (glioblastoma multiforme) and U87MG (anaplastic astrocytoma). To investigate effects on cell-proliferation, the cells were treated with IGF-I or II (0.001-100 ng/ml). The cell viability was assessed by MTT-assay. For testing migratory effects, the Boyden-chamber-assay with different combinations of IGF-I or -II or fetal calf serum (FCS) as chemotactic agents was used. RESULTS: All cell lines expressed IGF-I- and IGF-II-receptor, whereas none of the cells expressed IGF-I or -II. IGFBP 2-6 were found in all cell lines. IGF-I treated cell lines T98G and 86HG39 showed a weak dose-independent enhanced proliferation compared to controls, whereas A172 did not respond. None of the investigated cell lines changed proliferation when treated with IGF-II. All IGF-I (100 ng/ml) treated cells showed increased migration compared to controls. This effect was markedly enhanced by supplementation with 0.5% FCS. Again, IGF-II had no effect. CONCLUSION: These data demonstrate that IGF-I modulates proliferation and strongly stimulates migration of glioma cell lines in vitro. PMID- 18507055 TI - The clinicopathological and prognostic significance of MUC-1 expression in Japanese gastric carcinomas: an immunohistochemical study of tissue microarrays. AB - BACKGROUND: MUC-1 is synthesized as a single polypeptide that then undergoes proteolytic cleavage, and is associated with the epidermal growth factor receptor tyrosine kinases. In malignancies, MUC-1 may function as an anti-adhesion molecule, but can also promote adhesion and presumably metastasis. MATERIALS AND METHODS: Expression of MUC-1, -2, -4 and -5AC was evaluated on tissue microarrays of gastric carcinomas (n = 237) and adjacent non-cancerous mucosa specimens (n = 89) by immunohistochemistry and compared with clinicopathological parameters and survival time of the patients. RESULTS: MUC-1 was found to be highly expressed in gastric carcinomas in comparison with noncancerous mucosa (p < 0.05) and positively correlated with depth of invasion, lymphatic and venous invasion, lymph node metastasis, UICC staging and MUC-4 expression (p < 0.05), but not with age, tumor size, MUC-2 or MUC-5AC expression (p > 0.05). Intestinal-type carcinomas showed more MUC-1 expression than their diffuse-type counterparts (p < 0.05). Kaplan-Meier analysis indicated that the cumulative survival rate of patients with no MUC-1 expression was significantly higher than those with weak, moderate or strong expression in gastric carcinomas (p < 0.05), but no difference was observed when tumors were stratified according to the depth of invasion (p > 0.05). Cox's analysis showed three independent prognostic factors, depth of invasion, lymphatic invasion and venous invasion, to affect the relationship between MUC-1 expression and prognosis. CONCLUSION: Up-regulation of MUC-1 expression may be involved in pathogenesis, invasion, metastasis and differentiation of gastric carcinoma. Altered expression might therefore be employed as an indicator of pathobiological behavior of gastric carcinoma. MUC-1 expression was found to be a prognostic factor for gastric carcinoma patients, albeit not independent of parameters of invasion. PMID- 18507056 TI - Tumor-specific cytotoxicity and type of cell death induced by benzocycloheptoxazines in human tumor cell lines. AB - Twenty-six benzocycloheptoxazine derivatives were investigated for their tumor specific cytotoxicity and apoptosis-inducing activity against three human normal cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF) and four human tumor cell lines (squamous cell carcinoma HSC-2, HSC-3, HSC 4, promyelocytic leukemia HL-60). Benzo[b]cyclohepta[e][1,4]thiazine [1] exhibited very weak cytotoxicity, whereas its 6,8,10-tribromo derivative [3] exhibited higher cytotoxicity and tumor specificity (TS = 5.6). 6H Benzo[b]cyclohepta[e][1,4]diazine [4] and its cation [5] exhibited no tumor specificity. Among eighteen benzo[b]cyclohepta[e][1,4]oxazine derivatives [6-23], 6,8,10-triboromo- [9], 6-bromo-2-methyl- [20], and 6-bromo-2-chloro- [21] derivatives showed the highest tumor-specific cytotoxicity (TS = 12.5, 9.1 and 11.5, respectively). 14H-[1,4]Benzoxazino[3',2':3,4]cyclohepta[1, 2 b][1,4]benzoxazine [24] and its 7-bromo- [25] and 7-isopropyl- [26] derivatives had much lower cytotoxicity and tumor-specificity. Compounds [9, 20, 21] at 50% cytotoxic concentration (CC50) induced internucleosomal DNA fragmentation and caspase activation in HL-60 cells. On the other hand, these compounds induced apoptosis only at concentrations higher than CC50 in HSC-2 cells and failed to induce apoptosis in HSC-4 cells. Compounds [9, 20, 21] induced the formation of acidic organelles as measured by acridine orange staining. Transmission electron microscopy demonstrated the induction of moderate enlargement of mitochondria, the endoplasmic reticulum and nuclear membrane, and the vacuolation of the endoplasmic reticulum and the presence of a number of lamellar body-like organelles. These results indicate the diversity of the type of cell death induced by benzocycloheptoxazine derivatives in human tumor cell lines. PMID- 18507057 TI - Immunohistochemical expression of P15 (INK4B) and SMAD4 in advanced gastric cancer. AB - BACKGROUND: P15 (a cyclin- D- kinase inhibitor) and SMAD4 (a signal transducer of the TGF-beta pathway) are two closely related proteins which may have an important role in gastric carcinogenesis. MATERIALS AND METHODS: Sixty-three gastric carcinomas were studied. P15 and SMAD4 immunohistochemical expression was assessed with the DCS114.1 and BC/B8 mouse monoclonal antibodies, respectively. The results were tested for correlation with several clinicopathological parameters and patient survival. RESULTS: Fifteen out of the 18 (83%) P15 positive tumors also exhibited preserved SMAD4 positivity (p = 0.001). Loss of P15 expression was more frequent in the intestinal type of carcinoma (p = 0.047). The survival rates were significantly higher for patients with reduced SMAD4 expression, in cases of well- or moderately differentiated tumors (p = 0.042). CONCLUSION: P15 gene silencing might be an important event in the tumorigenesis of gastric adenocarcinomas of the intestinal type. Smad4 may behave as a tumor promoter in low grade tumors. The observed association between P15 and SMAD4 expression supports the inductive role of Smad4 in p15 transcription. PMID- 18507058 TI - Detection of cell-free ERCC1 and thymidylate synthase (TS) mRNA in malignant effusions and its association with anticancer drug sensitivity. AB - BACKGROUND: The aim of this study was to characterize cell-free mRNA in malignant effusions and evaluate its correlation with the sensitivity of primary tumor cells to cisplatin and 5-fluorouracil (5-FU). PATIENTS AND METHODS: Malignant effusions were collected from 43 patients and ERCC1 and TS mRNA transcripts in both the supernatants and tumor cells were characterized by real-time quantitative PCR. The sensitivity of primary tumor cells to cisplatin and 5-FU was analyzed using an ATP-TCA assay. RESULTS: Cell-free ERCC1 and TS mRNA transcripts were observed in 42 out of 43 specimens, with variable levels comparing gene expression patterns in paired-tumor cells (p < 0.0001, p = 0.02, respectively). TS mRNA levels in the supernatants were inversely correlated with the in vitro sensitivity of cancer cells to 5-FU in the gastrointestinal group (p = 0.002). CONCLUSION: The cell-free mRNA transcripts in malignant effusions were highly detectable and cell-free TS mRNA in gastrointestinal cancer patients were strongly associated with the sensitivity of primary cancer cells to 5-FU in vitro. PMID- 18507059 TI - Gypenosides inhibited invasion and migration of human tongue cancer SCC4 cells through down-regulation of NFkappaB and matrix metalloproteinase-9. AB - Gypenosides (Gyp), components of Gynostemma pentaphyllum Makino, were found to induce suppression of human tongue squamous cell carcinoma SCC4 cell growth and induce apoptosis in response to overexpression of reactive oxygen species, calcium (Ca(+2)) and to decrease mitochondrial membrane potential in vitro. In this study, the effect of Gyp on cell migration and invasion of human tongue SCC4 cells was examined. SCC4 cells treated in vitro with Gyp migrated and invaded less than cells treated with phosphate-buffered saline (PBS) as a control. Gyp inhibited migration and invasion by down-regulating the production of RAS, NFkappaB, COX2, ERK1/2 and MMP-9 relative to PBS only. These results show that Gyp inhibits invasion and migration of human tongue SCC4 cells by down-regulating proteins associated with these processes, resulting in reduced metastasis. PMID- 18507060 TI - Genetic polymorphisms of carcinogen metabolizing enzymes are associated with oral leukoplakia development and p53 overexpression. AB - BACKGROUND: Genetic polymorphisms of carcinogen-metabolizing enzyme genes have been associated with the risk of oral squamous cell carcinoma and oral leukoplakia. The overexpression of p53 protein is the most common genetic alteration in head and neck cancer. In the present study the combined or isolated presence of glutathione S-transferases GSTM1, GSTT1, GSTP1, and the cytochrome P450 oxidases CYP1A1 and CYP2E1 polymorphisms and oral leukoplakia development in a Brazilian sample of individuals was investigated, together with the effect of these polymorphisms on p53 overexpression in the lesions. PATIENTS AND METHODS: The GSTM1, GSTT1 and CYP1A1 genotypes of 80 smoking patients with oral leukoplakia and 80 age and gender matched control subjects were studied by polymerase chain reaction (PCR), and CYP2E1 and GSTP1 polymorphisms by PCR and digestion. Immunohistochemical reactions were performed for p53 staining in paraffin embedded histological sections of oral leukoplakia lesions. RESULTS: The GSTM1 null genotype was associated with an increased risk of oral leukoplakia development, independently of the other genes (OR 2.10). The simultaneous presence of GSTM1 and GSTT1 null genotypes was associated with an increased risk of oral leukoplakia development, independently of the other genes (OR 4.36). The oral leukoplakia lesions of patients with the GSTT1 null genotype showed a 6-fold increased risk of p53 overexpression (OR 6.61). CONCLUSION: A positive association exists between the isolated or combined null genotype of GSTM1 and GSTT1 and oral leukoplakia development and the null GSTT1 genotype shows increased risk of p53 overexpression, in oral leukoplakia. PMID- 18507061 TI - Circulating microparticles in breast cancer patients: a comparative analysis with established biomarkers. AB - BACKGROUND: The aim of the present prospective case-control study was to evaluate the putative relevance of circulating microparticles (MP) as a biomarker in breast cancer patients. MATERIALS AND METHODS: Endothelial cell-(EMP) and leukocyte-derived MP (LMP) were determined by flow cytometry in breast cancer patients (n = 41) and healthy controls (n = 25) and compared to carcinoembryonic antigen (CEA), cancer antigen (CA)15-3 and von Willebrand factor antigen (vWF) levels by specificity-sensitivity profiles. RESULTS: LMP, CEA and CA15-3 levels differed significantly between breast cancer patients and controls, whereas EMP and vWF did not. The specificity-sensitivity profiles of LMP and CA15-3 were similar. CONCLUSION: Increasing levels of circulating LMP (CD45+), CEA and CA15-3 correlated with increasing tumor size, thus reflecting disease stage. LMP showed an equal specificity-sensitivity profile to the established marker CA15-3 and therefore might have the potential to become a new biomarker in breast cancer patients. PMID- 18507062 TI - Prognostic significance of Ki-67 antigen expression in non-Hodgkin's lymphomas. AB - BACKGROUND: Non-Hodgkin's lymphomas (NHLs) are malignant tumours of the lymphoid system. Various risk factors have been described which are helpful in diagnosing, monitoring of the clinical course and predicting survival time of the patients. Proliferative activity of the tumour, measured by expression of Ki-67 antigen, is linked to the tumour proliferation rate and represents a recognised prognostic index in various tumours. In this study, the prognostic and predictive value of Ki-67 expression in NHL was evaluated. PATIENTS AND METHODS: Expression of Ki-67 was examined using an immunohistochemical technique in archival paraffin-embedded sections taken from 56 previously untreated patients with diagnosed primary NHL. An attempt was made to test correlation between Ki-67 expression on one hand and clinical parameters of the patients and their survival on the other. RESULTS: Expression of Ki-67 antigen was noted in 75% of the tumour cases. In the group manifesting higher Ki-67 indices, survival of the patients was significantly shorter (p = 0.03). No significant correlation could be detected between Ki-67 antigen expression and clinical or pathological parameters of the patients. CONCLUSION: The results demonstrate that the most cases of NHL display the expression of Ki-67. Moreover, shortened survival was noted in patients with high expression of Ki-67. PMID- 18507063 TI - Silibinin restores paclitaxel sensitivity to paclitaxel-resistant human ovarian carcinoma cells. AB - BACKGROUND: Drug resistance and tumor metastasis are the main causes of treatment failure and mortality in cancer patients. Silibinin, a naturally occurring flavanone, has been shown to be a potent sensitizer for apoptosis induced by a variety of anticancer drugs. In this study, whether silibinin could overcome chemoresistance and reduce the invasiveness of A2780/taxol cells was investigated. MATERIALS AND METHODS: A2780 and A2780/taxol cells were treated with silibinin alone and in combination with paclitaxel. Cell viability was determined by MTT assay while apoptosis and cell cycle progression were assessed by flow cytometric analysis. Matrigel invasion assays assessed the invasive activity. Protein and mRNA levels influenced by the treatment were studied by Western blots and quantitative real-time PCR. RESULTS: Silibinin enhanced the sensitivity of A2780/taxol cells to paclitaxel, increased paclitaxel-induced apoptosis and G2/M arrest consistent with the down-regulation of survivin and P glycoproteins. A2780/taxol cells demonstrated a two-fold increase in invasiveness ability compared to A2780 cells, whereas the invasive potential was reduced dramatically by silibinin. CONCLUSION: These results suggest silibinin in combination with paclitaxel may be a beneficial chemotherapeutic strategy, especially in patients with tumors refractory to paclitaxel alone. PMID- 18507064 TI - Surgical stress and accelerated tumor growth. AB - BACKGROUND: Delay in the initiation of radiotherapy after surgery is associated with an increase in local regional recurrence. A possible mechanism might be that remaining tumor cells proliferate significantly faster as a result of induced angiogenic cytokines. The growth rate of tumors arising from the inoculation of L44 tumor cells in the wound bed after surgical removal of L44 tumors was determined. MATERIALS AND METHODS: L44 tumors growing in the flank of female BN rats were surgically removed. In the wound bed, 5x10(6) L44 cells, harvested from the in vitro cell line, were injected. L44 cells were also injected in the contralateral flank and in control rats with and without surgical intervention. Tumor volumes as a function of time after injection of cells were recorded. From the attained volume at day 7, the cell doubling time was calculated, assuming 10(9) cells per cm3. RESULTS: Tumors arising in the wound bed had the fastest growth rate as compared to the tumors in the contralateral flank or tumors in control rats with or without surgical intervention. CONCLUSION: The results clearly indicate accelerated tumor growth after surgical stress. This indicates that delay in the initiation of radiotherapy after surgery with tumor cells remaining, results in a larger tumor burden and hence a higher probability of local recurrence. PMID- 18507065 TI - The vessel of life (about surgical reconstruction of the gastroepiploic artery). PMID- 18507066 TI - Serum lipids in parents and siblings of children with Alagille syndrome: a pilot study. AB - BACKGROUND/AIMS: Alagille syndrome, a dominant inherited disorder, is characterized by cholestatic liver disease, associated to interlobular bile duct paucity combined with; cardiac, skeletal, ocular and facial abnormalities. Increased levels of serum lipids are present in more than 80% of probands. Parents and siblings of children with Alagille syndrome are often found to have a mild expression of this probable disease gene; it is not known if dyslipidemia occurs in parents and siblings of children with Alagille syndrome. The aim of the study was to investigate the lipid profile in sibs and parents of children with Alagille syndrome. METHODOLOGY: Four children with Alagille syndrome and 21 first degree relatives were studied. SETTING: A pediatric referral hospital. PERIOD: July-October 2005. DESIGN: cross-sectional. VARIABLES: Total, low-density, high density cholesterol and triglyceride. STATISTICS: chi2, Mann-Whitney U and Kruskal-Wallis. RESULTS: Probands had higher levels of total cholesterol, low density cholesterol and triglycerides than their siblings (p<0.05); however, no differences with their parents were observed (p>0.05). CONCLUSIONS: Dyslipidemia does not seem to be a phenotypic expression in first degree relatives of children with Alagille syndrome. The increased level of serum lipids observed in some of the parents is similar to the expected prevalence of hypercholesterolemia in the adult Mexican population. PMID- 18507067 TI - Saliva transit from the oral cavity to the esophagus in GERD. AB - BACKGROUND/AIMS: Normal esophageal acid clearance depends on the neutralization of acid by swallowed saliva, as well as on esophageal peristalsis. This study therefore aimed to investigate the association between saliva production and transport from the oral cavity to the esophagus and developing gastroesophageal reflux disease (GERD) METHODOLOGY: Twenty-eight consecutive patients with GERD symptoms by gastroesophageal reflux self-report questionnaires and 13 healthy volunteers were recruited in the study. After intravenous administration of 99mTc pertechnetate, anterior sequential imaging was performed every minute for 40 minutes. At 20 minutes after injection of radionuclide, a lemon candy was administered intra-orally to stimulate salivary secretion. Regions of interest (ROI) were selected on the individual oral cavity, the pharynx, and the upper esophagus and time activity curves were drawn for each of these. A time-activity curve in each ROI was subjectively graded, as zero to two-point. The sum of scores in 3 ROIs was considered as a saliva transit total score. RESULTS: The mean transit score of the oral cavity did not differ significantly between GERD patients and healthy volunteers (1.38 vs. 1.61), whereas significantly lower transit scores of pharynx and upper esophagus were found in GERD patients. A 0 point saliva transit score of pharynx and upper esophagus was significantly more frequently detected in GERD patients than in healthy volunteers. CONCLUSIONS: This new modification of saliva scintigraphy is able to evaluate the esophageal motility simply, without a test meal, and to detect impaired saliva transit between pharynx and upper esophagus in GERD patients non-invasively. PMID- 18507068 TI - NOTES--A new era? AB - BACKGROUND/AIMS: After the first reports from the United States and India of accessing the peritoneal cavity via a transgastric route and performing operations without any abdominal incision, surgeons, as well as gastroenterologists worldwide, became interested in developing research projects in this topic. We evaluated the first papers and reports about the research and new techniques to focus on the possible advantages of NOTES (Natural Orifice Translumenal Endoscopic Surgery). METHODOLOGY: The literature was screened in the time period January 2000 to June 2007 for research and development in NOTES and several reports and abstracts from the year 2007 (January to June) were reviewed. RESULTS: Several research groups in the U.S. and Europe have published in this field of research and their advances and results are discussed. CONCLUSIONS: NOTES is a new era in surgery, but it will only partially replace laparoscopy as it will not be suitable for all patients and indications. To make NOTES suitable in daily surgical practice, it will take several years of research. NOTES research will boost the development of new endoscopes and instruments also helping to advance laparoscopic techniques. PMID- 18507069 TI - Contrast-enhanced endoanal and transperineal sonography in perianal fistulas. AB - BACKGROUND/AIMS: To improve the diagnostic potential of ultrasound in patients with perianal fistulas, by performing a combined perineal and endorectal Doppler sonography investigation before and after contrast enhancement with hydrogen peroxide. METHODOLOGY: Ten patients (9 male and 1 female) with perianal fistulas were tested. After contrast-free transperineal and endoanal sonography, hydrogen peroxide was injected in the fistulas and the ultrasound examinations were repeated. RESULTS: All fistulas were identified by conventional endoanal and transperineal sonography. The contrast application gave more detailed information about the location of the fistula in relation to the anal lumen and sphincters. The fistulas were distributed as follows: intersphincteric - in 5 patients, transsphincteric - in 2 and extrasphincteric complicated (with 1 or more secondary tracks) fistula - in 3 patients. The findings have been confirmed intraoperatively in 9 patients that underwent an operation. Hypervascularization of the wall was detected in 4 patients by means of Doppler investigation which also differentiated arterial blood flow in 3 cases and venous blood flow in one. CONCLUSIONS: The application of the contrast-enhanced endoanal and transperineal sonography offers a more detailed perianal fistula imaging. In addition the Doppler technique can further characterize their blood supply. PMID- 18507070 TI - Magnetic resonance cholangiopancreatography: potential usefulness of dehydrocholic acid (DHCA) administration in the evaluation of anastomotic site. AB - BACKGROUND/AIMS: The aim of this study was to investigate whether exogenous dehydrocholic acid (DHCA) was useful to enhance the delineation of anastomotic site. METHODOLOGY: DHCA is a cholagogue which produces an immediate effect by acting directly on liver cells. Its choleretic effect is strong, appearing 1 to 3 minutes after intravenous injection, reaching the maximum level in 20 to 30 minutes. Our study population comprised 9 patients. Magnetic resonance cholangiopancreatography (MRCP) was acquired before and after the administration of DHCA. Two different MRCP snap-shot techniques were applied: thick-slab two dimensional (2D) (coronal) single-shot turbo spin echo T2-weighted sequences and multisection thin-slab, 2D (coronal) single shot turbo spin echo T2-weighted sequences with three-dimensional (3D) maximum intensity projection (MIP) post processing. RESULTS: DHCA provided a better visualization of the anastomotic site in 7 patients (77.8%). The two patients without improvement in visualization of anastomotic site included 1 patient with liver cirrhosis secondary to portoenterostomy for congenital biliary dilatation and 1 patient, who was not eligible for the evaluation because of motion artifact caused by the difficulty of breath holding motion artifact. CONCLUSIONS: It was suggested that administration of DHCA could enhance the delineation of the anastomotic site on MRCP images. PMID- 18507071 TI - Sex-based differences in gallbladder cancer associated with pancreaticobiliary maljunction. AB - BACKGROUND/AIMS: Gallbladder cancer predominantly affects women; this sex-based difference is influenced by factors such as gallstones, sex hormone and genetic susceptibility. Gallbladder cancer is also frequently associated with pancreaticobiliary maljunction (PBM) without biliary dilatation. We examined sex based differences in gallbladder cancer associated with PBM. METHODOLOGY: With a particular focus on gender differences, we retrospectively compared clinicopathological findings between 44 patients (9 men, 35 women) with PBM without biliary dilatation and 232 patients (60 men, 172 women) with gallbladder cancer that was not associated with PBM. RESULTS: Gallbladder cancer was detected in 75% of patients with PBM without biliary dilatation. Among PBM patients, gallbladder cancer was significantly more common in women than in men [29/35 (83%) us. 4/9 (44%), p<0.05]. Both men and women with gallbladder cancer associated with PBM were significantly younger at the time of diagnosis than patients with gallbladder cancer without PBM (p<0.01). Gallstones were detected in only 10% of women with gallbladder cancer with PBM, while gallstones were detected in 63% of women with gallbladder cancer without PBM (p<0.01). CONCLUSIONS: Gallbladder cancer occurs very frequently in patients with PBM without biliary dilatation, and women appear to be at a significantly higher risk than men. PMID- 18507072 TI - Pancreaticoduodenectomy for bile duct carcinoma of a patient undergoing hemodialysis. AB - A 69-year-old Japanese man underwent pancreaticoduodenectomy for the resection of carcinoma at lower part of the common bile duct. Hemodialysis had already begun to treat chronic renal failure. He had been admitted for obstructive jaundice due to the carcinoma four months earlier. The serum total bilirubin was then 38.5 mg/dL, and the serum creatinine was 7 mg/dL. Hemodialysis was performed the day before the operation, and on the 1st, 3rd, 5th, 7th postoperative day. A rapidly degrading synthetic protease inhibitor was used as an anti-coagulant in the dialyzer to prevent systemic bleeding during the first postoperative week. Heparin was used from the second week. The maximum discharge from the drains was 2,300mL on the 3rd postoperative day. The drip intravenous infusion was changed from 1,900mL to 3,300mL during the first week to maintain the same body weight as the preoperative weight. Fresh frozen plasma and partial plasma fraction were used to maintain the colloidal pressure in the vessels so body weight reflects the fluid volume in the vessels. The postoperative course was uneventful. We present herein a successful case of pancreaticoduodenectomy for a patient undergoing hemodialysis to maintain the same body weight. PMID- 18507073 TI - Laparoscopic microwave ablation of liver tumors: our experience. AB - BACKGROUND/AIMS: Microwave ablation is the most recent development in the field of tumor ablation and is a well established and safe local ablative method available for liver tumors (both primary and secondary tumors). The technique allows for flexible approaches to treatment, including percutaneous, laparoscopic, and open surgical access. Laparoscopic technique has the advantages of accurate tumor staging, better tolerability and low cost. It can be performed in tumors which are close to the vital organs. The aim of this study was to evaluate the feasibility and safety of laparoscopic microwave ablation of liver tumors. METHODOLOGY: During January 2001 to December 2005, 57 patients with liver tumors were treated with laparoscopic microwave ablation in the department of Surgical Oncology. There were 34 male and 23 female patients. Out of 57 patients, 11 patients had hepatocellular carcinoma and 46 patients had secondaries in the liver. The most common source of secondaries was colorectal cancers. Laparoscopic microwave ablation of tumors was performed in these patients. RESULTS: During the study period, 57 patients with no evidence of extrahepatic disease underwent laparoscopic microwave ablation of unresectable hepatic tumors. No major intraoperative complications occurred. Postoperatively all the patients did well. Four patients developed liver abscess at the ablation area. Two patients required percutaneous aspiration of the liver abscess. No other major complications occurred. Follow-up CT scan shows complete necrosis of the tumors. Patients were followed-up at regular intervals. CONCLUSIONS: Laparoscopic microwave ablation is a feasible and safe alternative to open microwave ablation of the liver tumors. It carries all the advantage of minimal invasive surgery. In experienced hands, microwave ablation using laparoscopic technique can be done safely and effectively. PMID- 18507074 TI - An experimental study of emodin assisted early enteral nutrition for the treatment of severe acute pancreatitis. AB - BACKGROUND/AIMS: Both emodin and early enteral nutrition (EEN) have been affirmed as effective means to restore the intestinal mucosal function and abate the severity of severe acute pancreatitis (SAP). However, whether a combined strategy applying both is more effective than either one alone is still undetermined. In this study, we investigated the feasibility and efficacy of emodin assisted early enteral nutrition (EAEEN) for the treatment of SAP. METHODOLOGY: Sixty male Wistar rats were randomly divided into four groups (n=15). SAP was induced in all the rats by a retrograde infusion of 5.0% sodium taurocholate into the pancreatic main duct. Rats in group A received no further intervention, group B with emodin alone, group C with early enteral nutrition (EEN) alone, and group D with emodin assisted early enteral nutrition (EAEEN), respectively, all through an enteral nutrition tube incubated after the induction of SAP. 72 hours after SAP induction, all surviving animals were sacrificed to collect blood and tissue samples for the following measurements: serum amylase, tumor necrosis factor alpha (TNF-alpha), angiotensin II (AngII) and maleic dialdehyde (MDA), intestinal mucosal secretory IgA (SIgA), pancreatic myeloper oxidase (MPO) activity, and the wet-dry weight ratio of pancreatic tissue (pww/dw). The severity of pancreatic destruction was analyzed by pathological grading and scoring. The severity of intestinal mucosal damage was assessed by the wet-dry weight ratio of ileum (iww/dw), plasma D-lactate and plasma endotoxin. RESULTS: The results of every index in group B, C and D were significantly better than those in group A (P<0.05). Compared with group B and C, group D had significantly reduced levels of serum amylase, TNF-alpha, Ang-II and MDA (P<0.05). Group D also had significantly lowered plasma D-lactate and endotoxin, and decreased pancreatic MPO activity (P<0.05). The pww/dw and iww/dw ratios were decreased, while the SIgA level increased in this group, both with statistical significance (P<0.05). Furthermore, group D had significantly better pancreatic pathologic scores over group B and group C (P<0.05). CONCLUSIONS: Our results suggested that EAEEN could obviously abate the severity of experimental SAP in rats. This combined strategy was rational, safe and more effective than either EEN or emodin used alone, and has a broad potential for future clinical application. PMID- 18507075 TI - The clinical meaning of mucin phenotype and Epstein-Barr virus infection in gastric cancer. AB - BACKGROUND/AIMS: Differentiated type adenocarcinomas producing gastric type mucin are receiving much attention because of their degree of clinical malignancy. Most Epstein-Barr virus (EBV)-associated gastric cancers are undifferentiated type, and correlate with gastric type mucin. We analyzed the clinical meaning of mucin phenotypes and the detection of EBV in gastric cancers. METHODOLOGY: The objects of study were 120 consecutive gastric cancer lesions, resected endoscopically (EMR group, n=54) or surgically (surgery group, n=66). The mucin phenotypes were determined using immunostaining for human gastric mucin (HGM), MUC2, and CD10. Changes in histological type within the lesions were examined. The presence of EBV was determined using in situ hybridization for EBV-encoded small RNA 1 (EBER 1). RESULTS: The incomplete intestinal phenotype accounted for 83% of the EMR group, and the gastric phenotype for only 13%. None of the EMR group lesions had changes in the degree of differentiation, and there was no EBER-1-positive lesion. In the surgery group, the gastric phenotype accounted for 29%, significantly more than in the EMR group (p=0.0363). The incomplete intestinal phenotype accounted for 64% of surgically resected lesions. Changes in the degree of differentiation were significantly more common in the surgery group (16/66) than in the EMR group (0/54) (p=0.0001), tending to be more common in the gastric phenotype lesions. There were 3 EBER-1-positive lesions in the surgery group, accounting for 5%, and all were HGM positive. CONCLUSIONS: There appears to be little need to determine the mucin phenotype or EBV status of endoscopically resected lesions. In cases of gastric cancer where surgical resection is indicated, however, where the preoperative findings indicate a depth of invasion to SM or greater, and/or an undifferentiated lesion, then mucin phenotyping of biopsy specimens may be useful in predicting the predominant histological type of the tumor. PMID- 18507076 TI - Increase of serum CA19-9 level without an evident lesion on conventional imaging is insufficient for justification of 18FDG-PET examination. AB - BACKGROUND/AIMS: The serum tumor marker carbohydrate associated antigen 19-9 (CA19-9) has been used for screening for cancer, because its increase has been associated with many cancers. We aimed to evaluate the clinical value of positron emission tomography using F-18 fluorodeoxyglucose (18FDG-PET) that was prompted by increases of serum CA19-9 without findings on conventional imaging. METHODOLOGY: Twenty-two patients were retrospectively selected. Eleven were without a history of cancer and eleven had a history of cancer and were treated with curative intent. All 18FDG-PET findings were compared with the findings of histopathology by surgery or biopsy, or clinical follow-up for at least 1 year. RESULTS: We found only two true positive cases, and eleven cases without a cancer history included 10 true negatives and one false positive. CONCLUSIONS: Increases in serum CA19-9 are caused by many benign conditions. Increases of CA19-9 without findings on conventional imaging do not justify 18FDG-PET examination, particularly in patients without a cancer history. PMID- 18507077 TI - Association between serum gamma-glutamyltransferase and oxidative stress related factors. AB - BACKGROUND/AIMS: To investigate the main factors correlated with the serum gamma glutamyltransferase activity. METHODOLOGY: We measured serum gamma glutamyltransferase activity in 248 healthy Japanese people and determined its correlations with serum antioxidants, other plasma or serum factors, urinary 8 hydroxydeoxyguanine, and lifestyle factors. RESULTS: The mean serum gamma glutamyltransferase activity was 29 IU/L. Gamma-glutamyltransferase activities of males and persons older than 45 years were significantly higher than each counterpart. Gamma-glutamyltransferase levels increased significantly with the number of cigarettes smoked per day and the frequency of alcohol consumption except for the persons who did not take alcohol. Additionally, gamma glutamyltransferase significantly correlated with urinary 8-hydroxydeoxyguanine, and with more blood factors including serum tocopherols, carotenoids, antioxidative enzymes, lipid peroxide, and free fatty acids than urinary 8 hydroxydeoxyguanine did. In multiple regression analyses, gamma glutamyltransferase had significant associations with retinol, 8 hydroxydeoxyguanine, docosahexaenoic acid, and cigarette smoking. CONCLUSIONS: Our present findings support the hypothesis that gamma-glutamyltransferase can be used as a marker related with oxidative stress. PMID- 18507078 TI - Role of percutaneous transhepatic biliary drainage in patients with obstructive jaundice caused by local recurrence of gastric cancer. AB - BACKGROUND/AIMS: We reviewed the medical records of patients with obstructive jaundice caused by the local recurrence of gastric cancer to clarify the role of percutaneous transhepatic biliary drainage (PTBD). METHODOLOGY: Eleven patients with a mean age of 60.1 years (range: 51-71 years) underwent PTBD because of obstructive jaundice caused by the extrahepatic recurrence of gastric cancer. RESULTS: Jaundice was relieved in all the patients, and the serum total bilirubin (T-bil) level decreased from 12.2 to 2.1 mg/dL. No major complications associated with the execution of PTBD occurred. Although various symptoms caused by jaundice, such as anorexia, itching, nausea, abdominal pain, and fever, were relieved in all the patients within one week after PTBD, general fatigue persisted in 3 patients and abdominal fullness persisted in one. Seven of the 11 patients were discharged from the hospital after the execution of PTBD and remained at home for a median of 93 days. The median survival time (MST) of the remaining 4 patients who could not be discharged was 48 days. Chemotherapy was added in 5 patients after the execution of PTBD; these patients exhibited a significantly longer MST of 247 days, compared to 62 days among the patients who did not receive chemotherapy (P=0.0176). CONCLUSIONS: PTBD was safely conducted and improved the quality-of-life of patients with obstructive jaundice caused by the local recurrence of gastric cancer. Furthermore, the use of chemotherapy after PTBD might prolong patient survival although RCT (randomized controlled trial) study should be performed to assess the precise effect of chemotherapy after PTBD. PMID- 18507079 TI - Value of peroral cholangioscopy for mucin-producing bile duct tumor. AB - Mucin-producing bile duct tumor is a rare biliary tract tumor. Despite the development of modern diagnostic technologies such as ultrasonography, and angiography, the precise determination of this tumor is difficult because of ambiguity caused by the abundant mucin secreted by the tumor and/or by the superficial mucosal spread of the tumor along the bile duct. Given these problems, selective percutaneous transhepatic biliary drainage (PTBD) and percutaneous transhepatic cholangioscopy (PTCS) are indispensable for the accuracy staging of this cancer. But, PTCS has a risk of a rare but serous complications, seeding metastasis at the sinus tract of PTBD, whereas the retrograde approach minimizes this risk. We report the case of a patient who underwent the successful resection of a mucin-producing bile duct tumor. The tumor was diagnosed preoperatively as originating in the left caudate lobe after ERC and peroral cholangioscopy (POCS). PMID- 18507080 TI - Aberrant DNA methylation in ulcerative colitis without neoplasia. AB - BACKGROUND/AIMS: DNA methylation has been reported to correlate with the development of colitis associated cancer. We detected the promoter methylation of estrogen receptor gene (ER), TP53, p14, p16, p21 and hMLH1 in ulcerative colitis without neoplasia. METHODOLOGY: A total of 49 specimens from 36 patients, including 36 at rectal inflammatory mucosa and 13 at terminal ileum were obtained by colonoscopic biopsies. Methylation specific polymerase chain reaction were performed to detect the methylation in promoters of the above six genes. RESULTS: Methylation rate of ER promoter was significantly higher in the rectal mucosa than that in the ileum (76.3% vs. 46.2%, P=0.044). Moreover, ER methylation in rectal mucosa was significantly higher in relapse-remitting type compared to one attack only type cases (P=0.008), and also increased in cases longer than 7 years (P=0.036). Methylation rates of p14 or p16 were higher in rectal mucosa than those in the ileum, but the differences were not of statistic significance. Meanwhile, methylation in TP53 promoter was found in only one case, while p21 and hMLH1 methylation were negative in all cases. CONCLUSIONS: Methylation in promoters of ER, p14 and p16 occurs in rectal inflammatory mucosa without neoplasia. Examination of ER methylation in rectal mucosa may be useful for predicting cases at high risk of neoplasia. PMID- 18507081 TI - Evaluation and comparison of the clinical, surgical and pathological TNM staging of colorectal cancer. AB - BACKGROUND/AIMS: The aim of our study was to compare the results of clinical, surgical and pathological staging of colorectal cancer. METHODOLOGY: 660 patients with colorectal carcinoma were included in the study. The results of the clinical, surgical and pathological staging were compared. RESULTS: Clinical T values were identical with the surgical in 75.15%, and with the pathological in 74.54% respectively. Surgical T values were identical with the clinical in 78.48%. In 67.27% of the cases the clinical evaluation of N value was identical with the surgical one. Clinical evaluation was identical with the pathological result in 60.60% of the cases. Surgical diagnosis of the lymph node metastasis matched the pathological finding in 76.66%. Regarding the M value, the coincidence of the diagnoses was as follows: clinical versus pathological 72.72%, surgical versus pathological 90.90%. Clinical and surgical TNM stages were by 79.09% in accordance. By decision of total TNM stage the clinical-pathological staging showed worse (76.06%), while surgical-pathological showed significantly better (88.48%) matching. CONCLUSIONS: Based on our results we can state that in a quarter of all colorectal cancer cases the extent of the primary tumor could not have been established correctly. The lymph node involvement was well defined in just over half of the cases only. The M values were accurately stated in about three quarters of the cases. High grade of conformity of clinical, surgical and pathological staging can result in better treatment-planning, short- and long term survival, and higher quality of life. PMID- 18507082 TI - Polymorphism of peroxisome proliferator-activated receptor gamma is not associated to Japanese ulcerative colitis. AB - BACKGROUND/AIMS: Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) has been reported to be impaired in patients with ulcerative colitis (UC), and activation of PPARgamma is proved to inhibit the intestinal inflammation. As Plo12Ala polymorphism in codon 12 of the PPARgamma gene may decrease the promoter activity, we investigated the influences of PPARgamma polymorphism on the risk of UC in Japanese population. METHODOLOGY: The study recruited 118 patients with UC and 142 health controls. Plo12Ala polymorphisms of PPARgamma were detected by polymerase chain reaction based restricted fragment length polymorphism. RESULTS: The frequency of Pro/Alo heterozygotes of PPARgamma gene in UC and control group was 4.2% and 4.9%, respectively. No significant difference was found between UC and control group (P=1.00, Fisher's exact test). Plo12Ala genotype of PPARgamma did not show significant association with UC risk (OR=0.85, 95%CI=0.26-2.76). CONCLUSIONS: Our research suggests that Plo12Ala polymorphism of PPARgamma may not be associated with the risk of developing ulcerative colitis in Japanese population. PMID- 18507083 TI - Potential role of bcl-2 expression and apoptotic body index in colorectal cancer. AB - BACKGROUND/AIMS: Tumor growth is the result of proliferation and apoptosis. Bcl-2 is a proto-oncogene that inhibits apoptosis. The aim of the study was to investigate the clinicopathological correlations, interactions and prognostic significance of both bcl-2 protein expression and apoptotic body index (ABI) in colorectal cancer (CRCs). METHODOLOGY: Sixty colorectal cancer (CRC) patients had colonoscopic biopsies and tumor markers assay (CEA-CA19.9). The resected specimens were subjected for routine pathologic assessment, immunocytochemical staining for bcl-2 protein detection and immunofluorescence method for apoptotic body index. RESULTS: Bcl-2 immunostaining (IS) was positive in 29 patients (48.3%). The bcl-2 positive IS was significantly associated with -ve vascular invasion (P=0.05) and early tumor staging (P<0.01); the apoptotic body index had a median 3.5% and is neither correlated with bcl-2 or the clinicopathological variables. The overall survival (OS) was significantly associated with ABI (P<0.01), but not with bcl-2 expression and on bi-variant analysis, the OS is significantly better with high ABI in bcl-2 positive immunophenotype (P<0.05). CONCLUSIONS: Bcl-2 is a marker of favorable parameters (-ve angioinvasion and early tumor staging) but of no prognostic value. The apoptotic body index (ABI) is a favorable prognostic factor and may be used as a stratification parameter especially the high ABI in bcl-2 +ve CRCs. PMID- 18507084 TI - Management of rectal cancer: strategies and controversies. AB - Colorectal cancer is one of the most common cancers in the western world. The goal of this review is to outline some of the important surgical issues surrounding the management of rectal cancer. In patients with early rectal cancer (T1), local excision may be an alternative approach in highly selected patients. For more advanced rectal cancer, radical surgical resection is the treatment of choice. Total mesorectal excision and negative radial margin (>1 mm) decreases the local recurrence rate and improves survival. In appropriate patients, laparoscopic resection allows for improved patient comfort, shorter hospital stays, and earlier returns to preoperative activity level. In patients with locally advanced disease, neoadjuvant chemoradiotherapy followed by radical excision according to the principles of TME has become widely accepted. Surgical resection is the treatment of choice for resectable liver metastasis of colorectal origin. Surgical resection improves disease-free and overall survival rate. For patients with unresectable metastatic disease, multimodality approach may increase the resectability rate and hence survival. PMID- 18507085 TI - Possible role of human papilloma virus infection in response to neoadjuvant therapy in patients with esophageal cancer. AB - BACKGROUND/AIMS: Human papilloma virus (HPV) infection in esophageal cancer cases has been found in 0-70%, depending on different methods and geographical variances. Complete pathological response has been found in 30% of cases after neoadjuvant chemo-radiation (CRX). The aim of this study was to discover a possible relation between HPV-infection and response. METHODOLOGY: DNA was obtained from 26 esophageal cancer patients undergoing CRX and surgery. Polymerase chain reaction (PCR) and Southern Blot hybridization was used to detect HPV-infection (HPV-16 and -18). Clinicopathological parameters, disease free survival and overall survival were also analyzed. RESULTS: Complete response (26.9%) and partial response (38.5%) after CRX was correlated significantly with better prognosis. Six patients had HPV-infection (3 from the CR- and 3 from PR group). CONCLUSIONS: There was correlation between HPV-infection and response, but further analyses are necessary. Both responder-groups had a significantly better prognosis than non-responders. PMID- 18507086 TI - Predictive factors for anastomotic leakage in the neck after retrosternal reconstruction for esophageal cancer. AB - BACKGROUND/AIMS: Anastomotic leakage after transthoracic esophagectomy for esophageal cancer can induce life-threatening morbidity. This study investigated the predictive factors for anastomotic leakage in the neck after retrosternal reconstruction. METHODOLOGY: A total of 129 esophageal carcinoma patients undergoing transthoracic esophagectomy and esophagogastric anastomosis in the neck via a retrosternal approach were enrolled between April 1985 and March 2002. Predictive factors for anastomotic leakage were statistically evaluated. In a preliminary study using 18 cases, thoracic inlet space (TIS) was recommended to be extended more than 700 mm2. RESULTS: Partial resection of the bony structures was performed in 32 patients. The method of anastomosis and partial resection of bony structures according to the TIS independently influenced the likelihood of anastomotic leakage, with hand-sewn anastomosis and an absence of partial resection increasing its occurrence. CONCLUSIONS: Stapled anastomosis following the partial resection of the sternum and the left clavicle is recommended to avoid anastomotic leakage. These findings should be clarified by a randomized controlled study in a high-volume hospital. PMID- 18507087 TI - Adenocarcinoma of the esophagogastric junction in Japan. AB - BACKGROUND/AIMS: The prognosis of adenocarcinoma of the esophagogastric junction is worse than that in adenocarcinoma of other parts of the stomach. In particular, the clinical features and prognosis of adenocarcinoma of the esophagogastric junction and the differences between Siewert's type II and III tumors in Japan were evaluated. METHODOLOGY: We analyzed one hundred and forty patients with adenocarcinoma of the esophagogastric junction including one patient with a type I tumor, sixty-seven patients with type II tumors, and seventy-two patients with type III tumors. RESULTS: The prognosis of patients with type III tumors was poorer in comparison to that of type II tumors in adenocarcinoma of the esophagogastric junction (p<0.05). A significant difference was observed in the survival of patients with type III tumors between those with positive and negative lymph nodes (p<0.001). However, there was no such difference in patients with type II tumors. In a multivariate analysis, lymph node metastasis, age and the depth of tumor invasion were all found to be independent prognostic factors. CONCLUSIONS: The prognosis of patients with lymph node metastasis of type III adenocarcinoma of the esophagogastric junction was found to be extremely poor. An aggressive treatment after surgery may therefore be necessary to improve the survival of this population. PMID- 18507088 TI - Fibrous ring formation in cirrhotics treated with band ligation. AB - BACKGROUND/AIMS: We observed that the formation of a fibrous ring following variceal eradication appeared to be associated with less variceal recurrence. We aimed to evaluate this formally. METHODOLOGY: Twenty-one cirrhotic patients with a fibrous ring formation in the esophagus after eradication of varices (FR group) were compared with 21 controls of similar age, gender and liver function but without ring formation after eradication in terms of variceal recurrence, portal hypertension related bleeding and survival. RESULTS: Both groups were similar with regard to baseline demographic and clinical data. During a mean follow-up period of 28.8+/-18.3 (SD) months, variceal recurrence occurred in 2 (9.5%) patients in the FR group compared to 10 (47.6%) in the control group (p=0.005). Cox regression model revealed a significant difference in probability of variceal recurrence between the two groups (p=0.006). In the FR group 1 patient bled and 3 died vs. 2 and 6 patients in the control group respectively. The differences between the groups in relation to bleeding and death were not statistically significant. CONCLUSIONS: In cirrhotic patients undergoing band ligation for eradication of esophageal varices, the formation of a fibrous ring is followed by a lower variceal recurrence rate. PMID- 18507089 TI - Prediction of clinical response to anti-TNF treatment by oral parameters in Crohn's disease. AB - BACKGROUND/AIMS: The aim of our work was to predict the clinical response of Crohn's disease (CD) patients to anti-tumor necrosis factor (TNF) treatment by dental, periodontal, and oral mucosa parameters. METHODOLOGY: In 5 patients with luminal CD and in 9 patients with fistulizing CD, oral symptoms and signs before drug administration were assessed by 27 parameters, including: a) Decayed-, missing-, filled -, healthy -, nonvital -, root canal filled -, total number of - and impacted teeth; b) Oral ulcers, swelling of lips and cheeks, recurrent oral aphthae and hyperplasia of the mucosa; c) Width of keratinized gingiva, probing depth, gingival margin, clinical attachment level, bleeding on probing, plaque index, gingival index and gingival crevicular fluid (GCF) volume; d) The percentage of 8 morphotypes of subgingival plaque detected by darkfield microscopy (DFM), with the total number of bacteria examined. A new statistical prediction method has been introduced. RESULTS: At time lapse of 8 weeks after infliximab administration the prediction quality estimated from 8 optimal parameters attained a value of 0.63. However, with time lapse increasing to 3 months, the prediction quality decreased to 0.21, indicating that oral and response parameters became more statistically independent. CONCLUSIONS: This indicates that the oral state parameters could provide a sound basis for predicting the response of patients to infliximab administration. PMID- 18507090 TI - Duodenogastroesophageal reflux: results of medical treatment and antireflux surgery. AB - BACKGROUND/AIMS: Recent studies have shown that reflux of the duodenal content to the esophagus plays an important role in esophageal mucosal damage. The aim of the study is to compare the duodenogastroesophageal (DGER) reflux with the severity of reflux esophagitis and evaluate its response to either medical and/or antireflux surgery. METHODOLOGY: Ninety-six patients with DGER were subjected to thorough history, upper GI endoscopy, barium study, esophageal manometry and 24 hr esophageal pH metry combined with Bilitec 2000. Medical treatment was given for all, while Nissen fundoplication was done for 28 patients. All patients were evaluated after Nissen fundoplication and treatment. RESULTS: The age of studied patients was 36.26+/-12.7 years with male to female ratio 2:1. The chief symptom was heartburn in 73 (76%) patients. Upper GI endoscopy revealed, 30 (31.2%) patients had grade I reflux, 30 (31.2%) patients had grade II reflux, 7 patients had grade III reflux, 5 patients had grade VI reflux, Barrett's esophagus in 14 patients (14.5%), hiatus hernia (HH) in 26 (27%) patients. Barium study revealed that, 40 (41.6%) patients had evidence of reflux, while 34 (35.4%) patients had reflux with HH. Esophageal motility revealed the mean LESP (12.7+/-7.6), 68 patients (70.8%) had normotensive body while ineffective esophageal body motility was encountered in 28 (29.1%) patients. Esophageal 24-hr pH study and Bilitec 2000 revealed that 54 (56.2%) patients had bile reflux with pathological acid reflux, while 42 (43.7%) patients had bile reflux in alkaline pH. Medical treatment gave excellent to good response in 68 (70.8%) patients, while Nissen fundoplication was done for 28 (29.2%) patients. Endoscopic examination 6 months after Nissen fundoplication showed marked improvement in endoscopic injury. Barium study after Nissen fundoplication revealed repair of HH and control of GERD in all patients except one. Esophageal motility, 24 hr pH study and Bilitec 2000, after 6 months of Nissen shows high significant increase in LESP, decrease in acid and bile reflux. No significant difference between open or laparoscopic fundoplication in LESP, acid and bile reflux. CONCLUSIONS: DGER in acid medium is more injurious to the esophagus than DGER in alkaline pH. The severity of esophageal injury does not correlate with the severity of acid or bile reflux but has a direct correlation with impaired distal esophageal motility. Medical treatment gives satisfactory control of symptoms and healing of esophageal lesion in 70% of DGER. The response to medical treatment does not depend on the severity of esophageal injury but depends on the severity of bile and acid reflux. Nissen fundoplication in refractory patients, either open or laparoscopic, was effective in control of heartburn in 95% of patients contrary to 50% in mixed symptoms. PMID- 18507091 TI - Clinical and imaging characteristics relating to surgical outcomes of perforated appendicitis. AB - BACKGROUND/AIMS: To study the characteristics of clinical findings and CT imaging of perforated appendicitis for predicting the outcome of patients who received immediate appendectomy for perforated appendicitis. METHODOLOGY: Thirty-eight patients with perforated appendicitis who received immediate appendectomy were retrospectively reviewed. During a median follow-up period of 1091 days, 13 patients had to be re-hospitalized owing to occurrence of complications relating to the immediate appendectomy. Accordingly, the patients were divided into two groups as either complication or non-complication group. The clinical characteristics and CT imaging of these two groups were compared. RESULTS: Those patients who delayed seeking medical advice were more prone to develop surgical complications after immediate appendectomy. CT imaging showing either fat stranding with remarkable fluid content or abscess indicates the presence of severe inflammation and is related to adverse surgical outcomes. Moreover, extraluminal appendicolith was more frequently found in the CT imaging of complication group. CONCLUSIONS: Patients with perforated appendicitis differ in their severity. Patients who seek medical advice late or have signs of severe inflammation or extraluminal appendicolith on their CT imaging are associated with more severe diseases and are prone to develop complications of surgery at this time and should be better treated conservatively. PMID- 18507092 TI - Double-balloon enteroscopy for the diagnosis and treatment of obscure bleeding, inflammatory bowel diseases and polyposis syndromes: we see more but do we know more? AB - BACKGROUND/AIMS: Our aim was to report our experience with the Fujinon EN-450 T5 therapeutic double-balloon endoscope (DBE) in the diagnosis of small bowel diseases. METHODOLOGY: Between August 2005 and October 2006, 52 DBE procedures were conducted on 47 consecutive patients (M/F: 22/25, age: 51.6 SD 19.5 years) presenting at our tertiary referral hospital (35 and 7 patients from oral and anal route, respectively; 5 patients from both). All procedures were performed using i.v. anesthesia, at our outpatient clinic. RESULTS: Indication suspected small-bowel bleeding in 28 patients, suspected/known inflammatory bowel syndrome (IBD) in 12 and polyposis/suspected neoplasia in 7. In obscure bleeding small bowel abnormality was found in 18 patients (64.3%) including angiodysplasias/erosions and one polypoid lesion. In suspected IBD, IBD was diagnosed in 2 out of 8 cases. In patients with polyposis syndromes, polyps were in two Peutz-Jeghers patients, while a further patient with suspected stenosis was diagnosed with primary adenocarcinoma. The average insertion length was app. 213cm. No severe complications were observed. CONCLUSIONS: Based on our experience DBE is a safe and useful method for evaluating and treating small bowel disease in selected patients with obscure bleeding, IBD or polyposis syndromes, however the clinical importance of minute lesions still needs to be determined. PMID- 18507093 TI - Persistent oxidative stress in the corpus mucosa is evoked by long-term treatment of H. pylori-infected patients with proton pump inhibitors. AB - BACKGROUND/AIMS: To examine the influence of long-term treatment of H. pylori infected patients with proton pump inhibitors (PPI) on the toxic oxidant production in the stomach. METHODOLOGY: Eight H. pylori-positive patients with gastric ulcer were enrolled, and tissue samples were obtained endoscopically from the antrum and corpus. The tissue contents of neutrophil myeloperoxidase (MPO) and oxygen-derived free radicals were quantified by ELISA and chemiluminescence (ChL) assay. The H. pylori density in the tissue specimens was scored by immunohistochemistry and the mucosal infiltration by neutrophils and monocytes was scored by histopathology. The effects of PPI on these parameters were evaluated by oral administration of lansoprazole (LPZ) at the dose of 30mg od for 8 weeks, followed by 15mg for 24 weeks. RESULTS: Eight-week treatment with LPZ significantly increased the corpus ChL by 400%, which remained unchanged at this level during the subsequent 24 weeks, but the antrum ChL decreased slightly following LPZ-treatment. No significant changes in the mucosal MPO activity or the histologically determined parameters of H. pylori density and neutrophil/monocyte infiltration were observed in either portion of the stomach. CONCLUSIONS: The mucosal oxidative stress level significantly increased in the corpus mucosa after long-term treatment with LPZ, which may be independent of inflammatory cell infiltration. PMID- 18507094 TI - Early vs. delayed closure of temporary covering ileostomy: a prospective study. AB - BACKGROUND/AIMS: Temporary loop ileostomies constructed to protect distal anastomoses are generally closed at 8 to 12 weeks, a period long enough to encounter stoma-related complications, which reduces the quality of life. Early closure may be considered to overcome these adverse effects. This prospective study was designed to investigate the reliability of early closure of loop ileostomies. METHODOLOGY: Fifty consecutive patients were sequentially distributed either to group A (delayed closure) or group B (early closure). Anastomotic integrity was examined before closure in all patients. RESULTS: There were 25 patients in each group who were comparable in terms of age, sex, comorbid conditions, primary pathology, and tumor stage. Early closure was achieved in 88% (n=22) of the patients in group B. Stoma-related complications were significantly greater in group A patients (44% vs. 16%) (p<0.05) while the complications following closure were similar in both groups (16% vs. 8%) (p>0.05). CONCLUSIONS: Early closure during the same hospital admission produces less stoma-related complications with similar accomplishment as the late closure. Early closure of temporary ileostomy is recommended in suitable patients without anastomotic complications. PMID- 18507095 TI - Laparoscopic-assisted resection of gastrointestinal stromal tumor in small intestine. AB - Gastrointestinal stromal tumors (GISTs) in the small intestine are relatively rare. We present a case of GIST arising from the small intestine, which could be resected under laparoscopic assistance. A 60-year-old male visited another hospital due to massive anal bleeding. Blood examination showed severe anemia. Although anemia improved after conservative treatment, the bleeding site was not identified and he was referred to our hospital for close examination. Abdominal CT showed a well-defined mass with homogeneous internal density in the left upper abdomen. On abdominal angiography, selective enhancement of the second jejunal artery revealed a tumor stain. Due to suspected GIST in the small intestine, laparoscopic-assisted resection of the jejunum was performed. Histopathological examination demonstrated low-grade malignant GIST. The laparoscopic procedure is considered to be useful as minimally invasive surgery for diagnosis and treatment of selected cases with GISTs in the small intestine. PMID- 18507096 TI - Effect of the herbal medicine Inchin-Ko-To for serum bilirubin in hepatectomized patients. AB - BACKGROUND/AIMS: Persistent hyperbilirubinemia is a symptom of postoperative liver failure after hepatectomy. We examined the effectiveness of a herbal medicine, Inchin-Ko-To (ICKT), on postoperative serum bilirubin levels in patients undergoing liver resection. METHODOLOGY: Patients were divided into two groups. ICKT group (n=50), 7.5g of ICKT was administered orally from three days before the operation and it continued after the operation. Control group (n=50), ICKT was not administered perioperatively. RESULTS: There was no significant difference between the two groups in postoperative serum total bilirubin levels, but in indirect bilirubin, the ICKT group showed significant decrease compared with the control group. Decreasing effect of postoperative serum bilirubin levels was much more conspicuous when a large amount of liver parenchyma was resected. In that subgroup of patients, serum all bilirubin subdivisions significantly decreased in the ICKT (n=12) group compared with controls (n=11). CONCLUSIONS: ICKT may be an effective and fresh agent in postoperative management of liver resection by its potent choleretic effect. PMID- 18507097 TI - Induction of specific cytolytic T lymphocytes using fusions of hepatocellular carcinoma (HCC) patient-derived dendritic cells and allogeneic HCC cell line. AB - BACKGROUND/AIMS: To assess the ability of hepatocellular carcinoma (HCC) patient derived dendritic cells (DCs) fused with allogeneic HCC cell line to activate autologous lymphocytes to generate specific cytotoxic T lymphocytes (CTL) in vitro. METHODOLOGY: DCs were obtained by culturing adherent peripheral blood mononuclear cells (PBMC) from HCC patients in the presence of 100 microg/L recombinant human granulocyte/ macrophage- colony stimulating factor (rhGM-CSF) and 20 microg/L interleukin-4 (rhIL-4) for 1 week in vitro. DCs were fused with allogeneic HCC cell line HepG2 cells using polythyleneglycol (PEG), and the fusion cells were designated as DCs/HepG2. By labeling DCs and HepG2 with green and red fluoresceins, respectively, the cellular fusion was examined under fluorescence microscope. The ability of DCs/HepG2 to stimulate proliferation and differentiation of autologous lymphocytes was assessed by MTT method, and the specific killing efficacy of DCs/HepG2-induced CTL against HepG2 was evaluated. RESULTS: HCC patient-derived DCs expressed a certain level of CD1a, HLA-DR, CD54, CD80 and CD86. Fluorescence microscopic examination demonstrated that co incubation of DCs and HepG2 in the presence of PEG lead to generation of DCs/HepG2. In the mixed lymphocyte reaction assay, DCs/HepG2 had a significantly greater ability to activate proliferation of autologous lymphocytes, as compared with DCs alone, DCs plus HepG2, HepG2 alone and medium control (P<0.05). The DCs/HepG2-activated CTL showed a potent specific killing efficacy against HepG2 cells. CONCLUSIONS: Fusions of HCC patient-derived DCs and allogeneic HCC cell line could efficiently stimulate autologous lymphocytes to generate tumor specific CTL in vitro. It might represent a promising approach of immunotherapy for HCC. PMID- 18507098 TI - Convenience of a tape-guiding technique in different types of hepatectomy. AB - BACKGROUND/AIMS: The liver hanging maneuver is widely used in right lobectomy to resect huge tumors and harvest living donors. The convenience of tape assistance in other types of hepatectomy is not well known. METHODOLOGY: Tape-guiding technique (TGT) was applied in 30 hepatectomies of different type between April 2003 and April 2006. The indications were liver carcinoma in 22 and living-donor in 8. Hepatectomies included right lobectomy, 14; left lobectomy with caudate lobectomy, 8; left lobectomy without caudate lobectomy, 2; lateral segmentectomy, 3; central bisegmentectomy, posterior segmentectomy, and superior dorsal partial resection, 1 each. A tape was placed in front of the inferior vena cava for right hepatectomy and left hepatectomy with caudate lobectomy. In other hepatectomies, the tape was positioned to be the target of parenchymal dissection. RESULTS: TGT was successfully performed in all 30 cases. Tape facilitated dissection by helping the surgeon maintain orientation, and traction on the tape flattened the parenchyma, making it easier to identify and manage vessels and ducts. With an assistant holding the tape, the surgeon's left hand was free, and ligation and suturing was easier and more secure. CONCLUSIONS: The TGT is a convenient technique that is applicable to different types of liver resection. PMID- 18507099 TI - Clinical usefulness of serum C-reactive protein in living donor liver transplantation. AB - BACKGROUND/AIMS: The typical inflammatory response is frequently absent and the clinical manifestations of complications are often vague in LDLT recipients. So the rapid, sensitive, simple and noninvasive laboratory test may be very useful in postoperative care for LDLT recipients. METHODOLOGY: Excluding 5 pediatric LDLTs and 8 CDLTs, in total 100 adult LDLTs from January 2004 to June 2004 were retrospectively analyzed. RESULTS: Seventy-three episodes (24.9%) of CRP elevation were related with infection and 22 episodes (7.51%) were related with acute rejection. Sensitivity and specificity of serum CRP in infection and ACR were 0.90 and 0.65, respectively. The serum CRP level showed significant rise in infectious complications. The median peak value was 12.6 mg/dL and the range was 0.5-40.7 mg/dL. And recipients accompanying septic feature showed higher level of peak CRP. The recipients with acute rejection also showed elevated CRP but modest elevation (1.5-8.7 mg/dL). There was no correlation between the severity of rejection and serum CRP level. And the serum CRP rises were more frequently observed than leukocytosis and fever in recipients with infection or rejection. CONCLUSIONS: Serum CRP is quite sensitive but non-specific marker for diagnosis of infection and ACR. Serum CRP was a more sensitive marker for infection and ACR than fever or leukocytosis. PMID- 18507100 TI - Over expression of transforming growth factor-alpha and epidermal growth factor receptor in human hepatic cirrhosis tissues. AB - BACKGROUND/AIMS: Transforming growth factor-alpha has 30% amino acid homology to epidermal growth factor and binds with the same membrane-bound receptor, epidermal growth factor receptor. The purpose of this study was to investigate the expression of transforming growth factor-alpha and epidermal growth factor receptor in human hepatic cirrhosis tissues. METHODOLOGY: Expression of transforming growth factor-alpha and epidermal growth factor receptor was evaluated by immunohistochemistry stain in sixty-three hepatic cirrhosis specimens and five normal liver specimens. RESULTS: The transforming growth factor-alpha and epidermal growth factor receptor expression rates were 84.1% (53/63) and 52.4% (33/63), respectively. These positive granules were brown and most common in cytoplasm or cell membrane of hepatocytes. There was prominently positive correlation between transforming growth factor-alpha and epidermal growth factor receptor (P<0.05, gamma=0.32). In five normal liver tissues, transforming growth factor-alpha and epidermal growth factor receptor were not detectable in hepatocytes and bile ducts. CONCLUSIONS: Hepatic cirrhosis might be under the autocrine regulation of transforming growth factor-alpha and its receptor, epidermal growth factor receptor. Increasing expression of transforming growth factor-alpha and epidermal growth factor receptor might be one of the important events in hepatic cirrhosis pathogenesis. Furthermore, transforming growth factor-alpha might play a role in morphogenesis and regeneration of intrahepatic bile ducts. PMID- 18507101 TI - A new classification system for liver metastases from colorectal cancer in Japanese multicenter analysis. AB - BACKGROUND/AIMS: Although numerous authors have reported various prognostic factors for liver metastases from colorectal cancer, there is not yet a general classification. METHODOLOGY: A total of 478 colorectal cancer patients from 18 institutes were studied. Prognostic factors were investigated using univariate and multivariate analyses. RESULTS: Independent prognostic factors for colorectal liver metastases were number of liver metastases, size of the largest liver metastases, mesenteric lymph node metastases (pN0/1: < or =3 lesions, pN2: > or =4 lesions), and extrahepatic metastases (EM0: absence of extrahepatic metastasis, EM1: presence of extrahepatic metastases). We defined the following classification system; Stage A: HT1 (< or =4 lesions and < or =5cm) and pN0/1, Stage B: HT2 (> or =5 lesions or >5cm) and pN0/1, or HT1 and pN2, Stage C: HT2 and pN2, HT3 (> or =5 lesions and >5cm) with any pN, or any HT and any pN with EM1. Five-year survival rates were 53.5% for Stage A patients, 25.4% for Stage B patients, and 5.8% for Stage C patients. Median survival time was 70.4 months, 31.4 months, and 17.2 months, respectively. CONCLUSIONS: Our classification was advocated to evaluate prognoses for liver metastases from colorectal cancer. It can help guide decision making in terms of liver resection and assessing patient prognosis. PMID- 18507102 TI - Immunohistochemical study on liver fibrosis in biliary atresia. AB - BACKGROUND/AIMS: Although advanced liver fibrosis is a critical complication in influencing the outcome of biliary atresia (BA), the mechanism is poorly understood. In adult hepatic disorders, the relationships between sinusoidal liver cells (SLC) such as hepatic stellate cells (HSCs), some growth factors and enzymes concerned with extracellular matrix (ECM) metabolism have been clarified, but are unknown in BA. This study aimed to investigate such relationships in BA. METHODOLOGY: Seventeen liver samples from 14 patients with BA were immunohistochemically examined using primary antibodies such as alpha smooth muscle actin (alphaSMA), transforming growth factor beta (TGFbeta), platelet derived growth factor (PDGF), matrix metalloproteinase (MMP)-1, MMP-2, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2. The degree of liver fibrosis and these immunohistochemical findings were compared and examined semiquantitatively. Ultrathin sections from two samples were also examined with electron microscopy. RESULTS: The immunoreactivity of alphaSMA and MMPs increased with the degree of liver fibrosis, whereas that of TGFbeta, PDGF, and TIMPs showed no difference in expression in groups with any degrees of fibrosis. The immunoreactivity of MMPs statistically significantly increased in fibrotic livers. Electronmicroscopically, HSCs had many filaments in their cytoplasm, showing myofibroblastic morphology. CONCLUSIONS: The present study gave a different result than other reports on adult liver fibrosis. Livers with BA may be in a predominant state of fibrolysis, indicating the presence of the similar process to recovery from liver fibrosis in adults. PMID- 18507103 TI - Hepatic fibrosis influences the growth of hepatocellular carcinoma. AB - BACKGROUND/AIMS: This study is designed to evaluate the hepatocellular carcinoma (HCC) growth under physical pressure due to fibrotic condition. METHODOLOGY: 206 patients with HCC who consulted our institute in the past 15 years were selected. The patients were separated into three or two groups according to liver function or hepatic fibrosis, respectively. The preoperative mean value of serum N terminal procollagen type III (PIIINP) was estimated as an index of hepatic fibrosis. RESULTS: 1) Poorly differentiated type ofHCC tumor was significantly larger than the other types in cases with good-to-moderate liver function or without fibrosis. By contrast, in patients with poor liver function or fibrosis, no significant difference was found on tumor diameter (TD). 2) By contrast, although proliferating cell nuclear antigen labeling index (PCNALI) in cancer tissue was correlated with tumor diameter by simple linear regression analyses, PCNALI was consistently and significantly high in keeping with histological progression regardless of the status of liver functional or presence or absence of fibrosis. 3) The mean value of TD in intrahepatic metastasis (IM) positive cases, 45.8+/-5.1 mm, was higher than the size in negative cases. And the mean TD in IM positive cases was significantly smaller in the cases with poor liver function and fibroses. 4). In patients with small tumor, IM was noted 34.5%, 37.0% and 43.8% in good, moderate and poor liver function group, respectively. PCNALI was significantly higher in cases with moderate-to-poor liver function group or hepatic fibrosis. CONCLUSIONS: In the cases with poor liver function or hepatic fibrosis, cell proliferation cycle might be always activated even if the tumor is small. PMID- 18507104 TI - Preoperative CT during angiography of the liver for colorectal liver metastases. AB - BACKGROUND/AIMS: We had previously reported that using CT during angiography as a preoperative evaluation improves the detecting rate of minute (less than 10 mm) metastases and non-recurrent rate in the liver. We here evaluate the impact of CT during angiography on the survival after putative curative liver resection in our series. METHODOLOGY: During the period between 1990 and 2000, single detector helical CT was used for preoperative examination. There were 85 patients who underwent the examination before hepatectomy for colorectal metastases. Among them, there were 37 patients who underwent preoperative evaluating CT during angiography, and 48 who did not. The survival curves of the patient with/without CT during angiography, after the first hepatectomy, were calculated. RESULTS: Although recurrences in the residual liver after hepatectomy were observed in 9 out of 37 patients with CT-during angiography and 19 out of 48 patients without, the actual 5-year survival rates of the patients with and without CT during angiography are 42.6 and 43.2%, respectively, after more than 5 years' follow-up period. There is no significant difference between them. CONCLUSIONS: The present data show that the benefits from the approach to improve the detection of minute liver metastases are limited for the survival of putative curative liver resection. PMID- 18507105 TI - Long-term effectiveness of radiofrequency ablation for hepatocellular carcinoma of 3.5 cm or less. AB - BACKGROUND/AIMS: Hepatocellular carcinoma is one of the leading causes of death for cirrhosis, and patients are often not eligible for surgery. To evaluate the effectiveness of radiofrequency ablation in single (less than 3.5cm in diameter) or multiple nodules (up to 3, sized less than 3cm) in respect of acceptability, applicability, primary ablation rate, local recurrence, complications, and long term patients outcome. METHODOLOGY: 184 hepatic nodules detected in 156 consecutive patients recruited from a local sonographic screening were treated. In 10.2% of patients under study, a laparoscopic or laparotomy guided technique was preferred to the percutaneous approach. Overall and tumor-free survivals were estimated by Kaplan-Meier method. For the multivariate analysis, the hazard ratios and their 95 percent confidence intervals were computed by Cox model regression analysis. RESULTS: No treatment-related deaths and a severe complication rate of 3.2% were recorded. Primary complete ablation was obtained in 83.7% of nodules (87.1% of patients), and in a significantly higher rate for nodules up to 2cm (91.3%; p<0.013). Acceptability was 100%, and eligibility was very high (156 out of 160 cases). Local recurrence rate at 1 and 3 years was 10% and 25% respectively. The overall 3- and 5-year survival rates after treatment were 69.3% and 34.6%. Higher survival rates were obtained in the Child A cirrhosis subgroup (p<0.03) after complete response (p<0.001) and in the absence of new lesions (p<0.023). CONCLUSIONS: Radiofrequency ablation has great acceptability and applicability, and is a safe and effective treatment to be used after sonographic screening for small hepatocellular carcinomas. PMID- 18507106 TI - Des-gamma-carboxy prothrombin (DCP) ratio is a useful prognostic tumor marker for single nodule hepatocellular carcinoma (HCC). AB - BACKGROUND/AIMS: Des-gamma-carboxy prothrombin (DCP) is measured by an enzyme immunoassay system with the monoclonal antibody MU-3. A novel DCP antibody named 19B7 recognizes a different epitope against the Gla domain of DCP measured by the MU-3 antibody. Therefore, it is possible that DCP variants can be measured with these two antibodies. The aim of this study was to elucidate the usefulness of the DCP ratio as a new prognostic parameter for patients with hepatocellular carcinoma (HCC). METHODOLOGY: One hundred and eighty-three patients with HCC were enrolled in the current study. The DCP ratio was calculated using the formula: DCP ratio = DCP level measured by the MU-3 (mAU/mL) / DCP level measured by the 19B7 (mAU/mL). RESULTS: There was no statistical correlation between DCP level measured by MU-3 antibody and DCP ratio. Clinical stage, tumor type, portal tumor thrombus and DCP were independent factors in the multivariate analysis for survival of 183 patients. In 67 patients with single nodule HCC, clinical stage and DCP ratio were independent factors in the multivariate analysis for survival rate. CONCLUSIONS: The most useful prognostic tumor marker was the DCP in the 183 HCC patients and the DCP ratio in single nodule HCC. PMID- 18507107 TI - Suitable blending method of lipiodol-cisplatin in transcatheter arterial embolization for hepatocellular carcinoma: evaluation of sustained release and accumulation nature. AB - BACKGROUND/AIMS: Recently, fine powder cisplatin (IA-call; Nipponkayaku, Japan) was released, but there is no detailed study on an appropriate blending method of lipiodol-cisplatin for transcatheter arterial chemoembolization. We evaluated the sustained release and accumulation nature of lipiodol-cisplatin for hepatocellular carcinoma. METHODOLOGY: We prepared three types of mixture: a suspension of lipiodol and cisplatin powder, an emulsion of cisplatin powder dissolved with contrast medium and lipiodol, and a suspension-emulsion that was a suspension of lipiodol and cisplatin powder emulsified with contrast medium. In a basic study, a cisplatin release test was performed. In a clinical evaluation, transcatheter arterial chemoembolization with 3 lipiodol-cisplatin mixtures that had sustained release was performed in 60 consecutive patients with hepatocellular carcinoma as a randomized controlled trial. The density ratio of the tumor and treated liver tissue was measured as the accumulation nature. RESULTS: The suspension-emulsion and emulsion with a 7:3 mixture of lipiodol and contrast medium, and the suspension had better sustained release. The accumulation nature of the suspension-emulsion and emulsion were higher than the suspension. CONCLUSIONS: The pattern efficiently accumulated on hepatocellular carcinoma was the suspension-emulsion and emulsion. We suggest that a suspension emulsion may be created more easily and is more suitable for clinical use. PMID- 18507108 TI - Influence of estrogen and androgen on the outcome of liver transplantation. AB - BACKGROUND/AIMS: To assess influence of sex hormone on outcome of orthotropic liver transplantation (OLT). METHODOLOGY: Adult female Wistar rats were used as donors and male Wistar rats as recipients. Two experimental series were established. The first series consisted of the orchectomized (ORC) group. 17beta estradiol (E2) treated-ORC group and recipient Sham-ORC group; the second series consisted of the ovariectomized (OVX) group, dihydrotestosterone (DHT)-treated OVX group and donor Sham-OVX group. Recipients were sacrificed on postoperative day 7 (POD 7); the survival rate (SVR), histomorphological damage score of liver, graft-recipient weight ratio (GRWR) and the levels of alanine aminotransferase (ALT), alkaline phosphatase (AKP) and albumin (ALB) on POD 7 were detected. RESULTS: The histomorphological damage score and the level of ALT and AKP in the E2-treated ORC group, OVX group and DHT-treated OVX group was significantly lower compared with their respective sham group (P<0.05). CONCLUSIONS: Estrogen in recipient rats was responsible for the observed beneficial effects of liver transplantation, but at the same time, influence on liver of estrogen for a long time decreased the adaptability of graft to environment change. In contrary, androgen had less influence than estrogen. PMID- 18507109 TI - Serum IL-18 concentration does not depend on the presence of HCV-RNA in serum or in PBMC. AB - BACKGROUND/AIMS: To establish if serum IL-18 concentration depends on presence of HCV-RNA in serum or in peripheral blood mononuclear cells. To relate serum levels of IL-18 to histological parameters of chronic hepatitis C. METHODOLOGY: IL-18 serum concentration was measured by ELISA in 92 patients with chronic hepatitis C (43 serum HCV-RNA positive and 49 serum HCV-RNA negative after treatment). In 13 patients from the latter group HCV-RNA persisted in peripheral blood mononuclear cells, while in 34 it was negative. Thirty-five healthy individuals were the control. RESULTS: IL-18 level was higher in 92 patients than in the control (77+/ 31pg/mL us. 58+/-34pg/mL; p=0.002). Significant difference was found between the control and HCV-RNA positive (80+/-29pg/mL, p=0.002), HCV-RNA negative (75+/ 32pg/mL, p=0.019), patients who eliminated HCV-RNA from serum and from PBMC (85+/ 36pg/mL, p=0.002). IL-18 concentration was higher in serum HCV-RNA positive than in serum HCV-RNA negative patients (p=0.019) and it was related to histological inflammatory activity (p=0.008). CONCLUSIONS: Serum IL-18 concentration is related to presence of HCV-RNA in serum, in PBMC and liver inflammation. Increased level of this cytokine in patients with undetectable HCV-RNA may result from occluded persistence of the virus in the liver. PMID- 18507110 TI - Splanchnic non-hepatic hemodynamics and metabolism during liver transplantation. AB - BACKGROUND/AIMS: The aim of this study is to compare the splanchnic non-hepatic hemodynamics and the metabolic changes during orthotopic liver transplantation between the conventional with bypass and the piggyback methods. METHODOLOGY: A prospective, consecutive series of 59 primary transplants were analyzed. Oxygen consumption, glucose, potassium, and lactate metabolism were quantitatively estimated from blood samples from the radial artery and portal vein, collected up to 120 minutes after graft reperfusion. Mean arterial pressure, portal venous pressure, portal venous blood flow, and splanchnic vascular resistance were also measured or calculated at postreperfusion collection times. RESULTS: There was a greater increase in portal venous blood flow (p=0.05) and lower splanchnic vascular resistance (p=0.04) in the piggyback group. Mean arterial pressure and portal venous pressure were similar for both groups. Oxygen, glucose and potassium consumption were higher in the piggyback group, but none of the metabolic parameters differed significantly between groups. CONCLUSIONS: In conclusion, the study detected a higher portal venous blood flow and a lower and splanchnic vascular resistance associated with the piggyback technique. After graft reperfusion, no difference in the splanchnic non-hepatic metabolic parameters was observed between the conventional with bypass and the piggyback methods of orthotopic liver transplantation. PMID- 18507111 TI - Clinical implications of antimitochondrial antibodies in type 1 autoimmune hepatitis: a longitudinal study. AB - BACKGROUND/AIMS: Autoimmune hepatitis (AIH) is a chronic liver disease characterized by the presence of antinuclear antibodies. However, antimitochondrial antibodies (AMA) and bile duct changes, which are the characteristics of primary biliary cirrhosis (PBC), can be detected in AIH patients. METHODOLOGY: Twenty patients with definite AIH were prospectively followed-up, and the serial changes in AMA profiles were determined. We also examined the correlations between these antibodies and histopathological findings in the liver. RESULTS: Of the 20 patients, 7 (35%) had bile duct injury, and 2 of these 7 patients also showed chronic nonsuppurative destructive cholangitis or ductopenia of interlobular bile ducts histopathologically. Serologically, 7 patients (35%) were positive for AMA at least once by immunoblotting during the follow-up periods. There were no significant differences in biochemical hepatobiliary indices, the presence of bile duct lesions, or the changes in biochemical profiles between AMA-positive and AMA-negative AIH patients during the follow-up periods. CONCLUSIONS: We confirmed that AMA and certain histopathological findings that are characteristics of PBC can be seen in some AIH patients. However, there was no significant correlation between AMA positivity and the histopathological findings in the liver, or biochemical hepatobiliary indices. Thus, the clinical implications of AMA in AIH patients remain unclear. PMID- 18507112 TI - Reconstruction of hepatic venous tributaries using a Y-shaped left portal vein graft harvested from a resected left liver. AB - Hepatic vein resection and reconstruction after major hepatectomy is a technically feasible but controversial procedure. Reported autologous vein grafts include the great saphenous, external iliac vein, superficial femoral, gonadal, left renal and inferior mesenteric veins. The procedures required to obtain these grafts, however, are associated with a risk of postoperative morbidity such as edema of the lower leg. We performed the reconstruction of two middle hepatic vein (MHV) branches by using an autologous left portal vein graft that was harvested with its tributaries from the left hepatectomy specimen in a 57-year old man who had undergone a sigmoidectomy for colon cancer and a partial resection of the right lateral sector of the liver for a metastasis. In conclusion, this is the first report on the reconstruction of MHV tributaries using a single autologous Y-shaped portal vein graft during a hepatectomy. This method produces no complications related to the harvesting of the graft. PMID- 18507113 TI - Growth inhibition of unresectable tumors induced by hepatic cryoablation: report of two cases. AB - The efficacy of percutaneous cryosurgery (PCS) as a treatment strategy for unresected liver tumor was evaluated in two cases. The first patient was a 64 year-old man who was found to have multiple liver tumors after undergoing gastrectomy for gastric cancer (T3, N0, M0, Stage II). Two PCS treatments under local anesthesia decreased the size of both the treated and untreated tumors. The second patient was a 61-year-old man in whom multiple liver tumors were discovered after hepatectomy for metastases of a duodenal gastrointestinal stromal tumor, which also had been treated surgically. A third surgery was performed for mass reduction. The patient showed stable improvement after surgery, and PCS combined with administration of polysaccharide-Kureha was selected to treat the unresectable tumors. PCS was performed once a week with an overnight hospital stay. After nine PCS treatment, the remarkable reduction in the size and number of liver tumors was observed, even among non-treated tumors. The patient remains in good condition without tumors 21 months after treatment. PMID- 18507114 TI - Following up of liver transplantation using dual left grafts from living donors- one case. AB - Living donor liver transplantation (LDLT) using right lobe graft was advocated to treat adult patients who could not receive a timely cadaveric graft. The major limitation of adult-to-adult LDLT using right lobe graft is the safety of the donors. Adult-to-adult LDLT using dual left grafts was performed for one patient with end-stage liver disease in our division. Both donors and recipient underwent a full recovery from the operation without persistent complications. Two left dual grafts from two living donors can help alleviate the pressure of organ shortage, prevent the small-for-size problem and yet secure the safety of the donors. PMID- 18507115 TI - Stent-graft to treat intrahepatic portosystemic venous shunt causing encephalopathy. AB - Spontaneous intrahepatic portosystemic venous shunts (IPSVS) are rare, and angiographic intervention to treat IPSVS has been limited. To our knowledge, we present the first case of an IPSVS between the right portal vein and inferior vena cava accompanied by intractable encephalopathy that was successfully obliterated using a stent-graft deployed in the inferior vena cava to disconnect the portal and systemic venous circulation. Our patient, a 58-year-old woman, has shown no evidence of recurrent portosystemic venous shunt or encephalopathy for 30 months. We believe this intervention to be a useful alternative therapy for this type of shunt. PMID- 18507116 TI - B-RTO for ectopic variceal bleeding after living donor liver transplantation. AB - Gastrointestinal bleeding (GIB) is a serious gastroenterological complication after orthotropic liver transplantation, as well as living donor liver transplantation (LDLT). Although ectopic varices are rare causes of GIB, they may lead to life-threatening bleeding and it is difficult to diagnose them. We herein report a rare case of a jejunal variceal bleeding in a recipient five years after LDLT for primary sclerosing cholangitis with successful result by balloon occluded retrograde transvenous obliteration (B-RTO). PMID- 18507117 TI - Transampullary pancreatic duct stenting decreases pancreatic fistula rate following left pancreatectomy. AB - BACKGROUND/AIMS: Significant improvements in mortality following pancreatic surgery have been noted by high-volume centers in recent years. Despite this, morbidity from pancreatic resection remains high, with postoperative pancreatic fistula remaining a common problem following distal pancreatectomy (DP). Rates of pancreatic fistula following distal pancreatectomy have ranged from 0 to 61% in a recent meta-analysis of surgical techniques and impact upon pancreatic fistula rates. We postulated that intraoperative placement of a transampullary pancreatic duct stent (TAPDS) at the time of distal pancreatectomy, would decrease ampullary complex-mediated elevation in pancreatic duct pressures, improve healing of the ligated pancreatic duct and result in a decrease in pancreatic fistula following distal pancreatectomy. METHODOLOGY: Sixteen consecutive patients underwent distal pancreatectomy plus TAPDS and were compared to 43 control patients who underwent distal pancreatectomy by the same surgeon, with identical management of the pancreatic remnant. Distal pancreatectomy was performed as the primary operation or as part of an en-bloc resection for a primary malignancy other than pancreatic adenocarcinoma. In patients who underwent transampullary pancreatic duct stenting (TAPDS), the pancreatic duct was identified after transection of the pancreatic parenchyma. A soft, pediatric feeding tube was inserted directly into the pancreatic duct and carefully fed into the duodenum (confirmed by palpation). The stent was placed distally, one centimeter from the cut-edge of the pancreatic duct, which was then ligated as described earlier. Closure of the pancreatic parenchyma was identical to those patients who did not undergo TAPDS placement. Common perioperative outcomes were assessed, including pancreatic fistula. RESULTS: No statistically significant differences where found between the rates of intraabdominal abscess, intraabdominal hemorrhage or need for reoperation. Pancreatic fistula rates and average length of stay were significantly decreased in patients undergoing distal pancreatectomy with TAPDS (p<0.05 and p<0.0001 respectively). CONCLUSIONS: Statistically significant reductions in pancreatic fistula and average length of stay were noted in patients who underwent stenting of the pancreatic duct with TAPDS. PMID- 18507118 TI - Pancreas divisum in pancreaticobiliary maljunction. AB - BACKGROUND/AIMS: Pancreaticobiliary maljunction (PBM) and pancreas divisum (PD) are congenital anomalies that develop in the embryo at an early stage. They are possibly the result of bile and pancreatic duct misarrangement. We investigated the configuration of the pancreatic duct in patients with PBM and its clinical implications. METHODOLOGY: In 84 PBM patients, the configuration of the pancreatic duct and the presence of biliary cancer were documented. Patency of Santorini's duct was determined fluoroscopically or by dye-injection endoscopic retrograde pancreatography. Bile amylase levels were measured in 10 patients. RESULTS: Incomplete PD was detected in 8 (9.5%) of the 84 PBM patients. All of the 8 patients had a patent Santorini's duct, and only 1 patient had gallbladder cancer. The frequency of associated gallbladder cancer and the bile amylase level were significantly lower in PBM patients with a patent Santorini's duct than in PBM patients with a nonpatent Santorini's duct. CONCLUSIONS: PBM is sometimes associated with incomplete PD. In PBM patients with an incomplete PD, the incidence of cancer of the biliary tract may be lower, since pancreatic juice reflux into the bile duct might be reduced by the flow of pancreatic juice into the duodenum through Santorini's duct. PMID- 18507119 TI - Significance of palliative gastrojejunostomy for unresectable pancreatic head carcinoma. AB - BACKGROUND/AIMS: When diagnosed, most patients with pancreatic head cancer are defined as having an unresectable tumor, and thus appropriate relief of main symptoms, such as, obstructive jaundice, duodenal obstruction, and pain, is the major concern. However, debate continues concerning the efficacy of gastrojejunostomy at preventing duodenal obstruction. In this study, we evaluated the effects of palliative gastrojejunostomy on the quality of life of patients with unresectable pancreatic head cancer. METHODOLOGY: Between January 2000 and December 2004, of 61 patients with unresectable pancreatic head cancer, 46 underwent palliative surgery; 24 patients underwent biliary bypass with gastrojejunostomy (Group I) and 22 patients underwent biliary bypass only (Group II). Frequencies of nausea and vomiting, serum protein and albumin levels, oral intake, and other metrics were carefully monitored. RESULTS: No significant differences in age, sex, TNM stage, and preoperative symptoms were evident between the two groups. Incidences of nausea and vomiting were similar preoperatively (Group I, 45.8%; Group II, 40.9%, P=0.736), but were significantly different at 3 months postoperatively (Group I, 25.0%; Group II, 68.2%, P=0.003). Preoperative serum protein and albumin levels were similar in the two groups (Group I: 6.6g/dL, 3.5g/dL; Group II: 6.4g/dL, 3.4g/dL, respectively, P-0.223, 0.472), but at 3 months postoperatively, serum protein and albumin levels were significantly different (Group I: 5.9g/dL, 3.1g/dL; Group II: 5.2g/dL, 2.6g/dL, respectively, P=0.010, 0.047). CONCLUSIONS: Our data suggest that palliative gastrojejunostomy in patients with unresectable pancreatic head cancer reduces symptoms related to duodenal obstruction and contributes to quality of life. PMID- 18507120 TI - Neopterin--a potential factor for differentiation between pancreatic cancer and chronic pancreatitis. AB - BACKGROUND/AIMS: Pancreatic cancer constitutes a difficult diagnostic and therapeutic problem. It remains the fourth cause of death among gastrointestinal malignancies. The present study aimed to investigate whether neopterin can be used as a significant marker in pancreatic cancer and to establish any possible relationships between serum neopterin levels and the differentiation between chronic pancreatitis, pancreatic cancer, and acute pancreatitis. METHODOLOGY: The research was done on a group of 65 patients with a diagnosed pancreatic tumor and on a group of 21 patients with a diagnosed acute pancreatitis. Serum neopterin concentration was determined in all patients on the first day of hospitalization. RESULTS: Based on intraoperative and postoperative histopathological findings, pancreatic cancer was diagnosed in 42 patients and chronic pancreatitis in 23 patients. Neopterin concentrations in pancreatic cancer patients, chronic pancreatitis patients, and acute pancreatitis patients were 13.01-29.64 nmol/L (mean 18.16), 0.95-11.81 nmol/L (mean 6.52), and 9.64-25.0 nmol/L (mean 13.61), respectively. Mean neopterin levels in pancreatic cancer and acute pancreatitis patients were higher than the accepted norm (norm<10 nmol/L). Serum neopterin concentrations in patients with chronic pancreatitis remained within the norm. CONCLUSIONS: In our opinion, neopterin might be a promising factor of differentiation between pancreatic tumors. However, the use of neopterin as a nonspecific tumor marker requires further studies. PMID- 18507121 TI - Laparoscopic endogastric and transgastric cystgastrostomy and pancreatic necrosectomy. AB - BACKGROUND/AIMS: Pancreatic pseudocysts (PP) that complicate acute necrotizing pancreatitis (ANP) and require internal drainage may be managed laparoscopically. We present our experience with the laparoscopic endogastric and transgastric approaches to pseudocyst-gastrostomy (PCG). METHODOLOGY: Seven patients (4 female) aged 25-75 (median, 60) years with large (median, 12cm), symptomatic and persistent (median, 5 months) retrogastric PP underwent PCG. All procedures were attempted laparoscopically. The endogastric approach was applied in the initial three patients and was replaced with the transgastric approach in the subsequent four patients. All patients underwent concomitant necrosectomy for sterile pancreatic necrosis. Patients were followed up with regular abdominal ultrasonography. Results shown represent median (range). RESULTS: There were no conversions to open surgery. The operating time was 135 (60-200) minutes and was longer with the endogastric approach (165 vs. 112 minutes). There were no postoperative complications and the postoperative hospital stay was 2 (1-4) days. There were no recurrences at a follow-up of 12 (2-21) months. CONCLUSIONS: Laparoscopic PCG with concomitant necrosectomy for PP that complicate ANP is feasible and safe and is associated with smooth and rapid recovery. The transgastric approach provides better access and is simpler to apply than that of the endogastric approach. PMID- 18507122 TI - Therapeutic tactics in the treatment of acute necrotizing pancreatitis. AB - BACKGROUND/AIMS: The authors analyze the possibilities for the delay of surgery with special consideration regarding percutaneous peripancreatic drainage in the treatment of acute necrotizing pancreatitis. METHODOLOGY: In addition to intensive care therapy, 61 patients were also given antibiotic prophylaxis, and early nasojejunal enteral feeding was commenced. In a total of 22 cases where peripancreatic fluid was found, percutaneous drainage was performed. Septic necrosis, sepsis, multi-organ failure not resolving with conservative treatment, gastrointestinal perforation, and bleeding were the indications for operation. Only 9 patients underwent surgery within one week and in 40 patients delayed (more than 7 days) necrectomy was performed. Following surgery, closed omental bursa rinsing was performed. RESULTS: Five patients were cured with only conservative therapy and 7 others were cured under the influence of percutaneous drainage. In 15 patients it was possible to delay surgery using percutaneous drainage with combination of conservative treatment. A total of 39 reoperations occurred due to septic focus, bleeding, colonic necrosis and gastric perforation. The average days of nursing care was 43.3 (3-120). Mortality was 16.4% (10/61 patients). CONCLUSIONS: The number of early operations can be reduced with the use of antibiotic prophylaxis, nasojejunal feeding and percutaneous drainage. PMID- 18507123 TI - Virtual CO2 MDCT pancreatography: a new feasible technique for minimally invasive pancreatectomy in intraductal papillary mucinous neoplasms. AB - BACKGROUND/AIMS: Less invasive pancreatic head resection, such as duodenum preserving pancreatic head resection (DPPHR) has been introduced for the treatment of pancreatoduodenal lesions, especially for benign conditions, for reducing surgical stress and maintaining exocrine and endocrine function of the residual pancreas in consideration of postoperative quality of life (QOL). METHODOLOGY: We investigated the feasibility of a new technique employing three dimensional (3D) virtual pancreatography using multi-detector CT (MDCT) with carbon dioxide (CO2) gas as a negative contrast agent for detection of intraductal papillary mucinous neoplasm (IPMN) of the pancreas requiring minimally invasive surgery. Branch IPMN is subjected in this study. RESULTS: Contrast-enhanced MDCT scan of the abdomen diagnosed 4- to 20-mm multilocular septated cysts in the head-uncinate process of the pancreas. Endoscopic retrograde pancreatography (ERP) showed multiple cystic lesions in the head uncinate process with mild dilatation in the remaining pancreatic duct. For localizing diagnosis of these small and multiple pancreatic cysts, we placed an endoscopic pancreatic stent (EPS), and MDCT with injection of CO2 via EPS was examined for the virtual CO2 pancreatography, consisting of OsiriX software system employing 3D virtual anatomic reconstruction with CO2 gas as a negative contrast agent. Virtual CO2 MDCT pancreatography demonstrated that all cystic lesions of the pancreas were contained within the area of the head-uncinate process of the pancreas. We performed DPPHR, and surgical margin of the patient's remnant pancreas was determined as non-malignant by intraoperative histology. There was no residual pancreatic cyst and tumor after surgery. The resected tumor was diagnosed as branch duct type intraductal papillary mucinous adenocarcinoma. According to our minimally invasive DPPHR obtained by virtual CO2 pancreatography, the pancreatic endocrine and exocrine functions of this patient were maintained at almost the same levels as those in his preoperative status. With respect to preservation of the endocrine and exocrine functions of the pancreas, DPPHR is a highly effective surgical procedure due to limited surgical resection. CONCLUSIONS: Our new technique of virtual CO2 MDCT pancreatography is a feasible procedure for preservation of the remnant pancreatic function. This is the first report of virtual CO2 pancreatography providing minimally invasive pancreatic surgery. PMID- 18507124 TI - Pancreatic neuroendocrine tumors: our approach. AB - BACKGROUND/AIMS: Pancreatic neuroendocrine tumors constitute a small percentage of pancreatic tumors. Surgical resection is the best treatment for these types of tumors. Aggressive surgical resection including multivisceral resection provides long-term survival. Even palliative resection of the tumor is justifiable. Here we share our experience with the management of pancreatic neuroendocrine tumors. METHODOLOGY: Between January 1993 and April 2007 we operated on 54 patients with pancreatic neuroendocrine tumor. We have analyzed our data retrospectively. Patients were analyzed in terms of demographic characteristics, operative procedure, postoperative outcome and survival. RESULTS: Out of 54 patients, 31 patients had nonfunctional tumor and 23 patients had functional tumors. Neuroendocrine carcinoma was found in 19 patients. Pancreaticoduodenectomy was performed in 21 patients. Simultaneous liver resection was performed in 4 patients and multiorgan resection for locally advanced pancreatic tumor was performed in 3 patients. CONCLUSIONS: Surgical resection is the best option for the treatment of pancreatic neuroendocrine tumors. Aggressive resection provides survival benefit and a better quality of life. If the entire gross tumor can be resected, multiorgan resection or simultaneous liver resection is justifiable. PMID- 18507126 TI - Management of congenital and posttraumatic splenic cysts in children. AB - Splenic cysts are rarely observed in young patients. We report three cases of splenic cysts. The management and the surgical procedure, if needed, are discussed. If the splenic cyst is symptomatic or if its size reaches a diameter of 4-6cm, it requires surgical management. This treatment would be performed by laparoscopic approach and is intended to spare splenic tissue as much as possible. A prophylactic anti-pneumococcal vaccination is recommended before surgical treatment. PMID- 18507125 TI - Usefulness of diffusion-weighted imaging (DWI) for the detection of pancreatic cancer: 4 case reports. AB - The clinical utility of diffusion-weighted magnetic resonance imaging (DWI) was originally established for acute stroke; however, recent studies suggest that DWI may be more sensitive and specific for the detection and staging of malignant tumors than either computed tomography (CT) or ultrasonography (US). We herein present 4 cases of pancreatic cancer that were detected by DWI and subsequently discuss the efficacy of DWI for the diagnosis pancreatic cancer. We performed both DWI and dynamic CT examinations on 4 patients with pancreatic cancer. MR examinations were performed with a 1.5-T imager (Toshiba). We measured the signal intensity in a series of DWI images and calculated the apparent diffusion coefficient (ADC) values to differentiate the tumors from normal tissue, inflammation, or another lesion. Two radiologists analyzed the DWI and CT images, and the evaluation of the primary tumor (T), regional lymph nodes (N), and distant metastatic disease (M) was conducted according to the TMN classification system. There were no differences between the DWI and the CT images regarding their abilities to detect advanced pancreatic cancer. However, DWI displayed superior ability in detecting early stage tumors and evaluating the degree of tumor invasion. DWI has the potential to be clinically effective in the detection of early pancreatic cancer, and DWI can be a powerful tool for the evaluation of pancreatic cancer. PMID- 18507127 TI - The prevalence of primary and secondary Helicobacter pylori resistance to clarithromycin and probable contributing cofactors: data from southeastern Anatolia. AB - BACKGROUND/AIMS: Antibiotic resistance of Helicobacter pylori is the most important reason for failure in its eradication. We aimed to determine the prevalence of primary and secondary H. pylori resistance to clarithromycin in isolated H. pylori from dyspeptic patients in southeastern Anatolia and to evaluate the cofactors affecting this clinical problem. METHODOLOGY: The study involved adult patients who had already been diagnosed with symptomatic H. pylori infection based on rapid urease test, gastric histopathological examination and culture. H. pylori strains were isolated from antral biopsies taken during upper endoscopy in 142 dyspeptic patients with no previous therapy against the microorganism. MICs of clarithromycin were determined by E-test. Patients were treated for 14 days with standard triple-agent protocol. H. pylori eradication rate was assessed after 8 weeks. Each patient was re-interviewed to determine secondary resistance. Primary clarithromycin resistance was defined as pre treatment resistance, while secondary as after treatment resistance. Strains were considered resistant to clarithromycin if the MIC > 1 microg/mL. RESULTS: In total 213-105 women and 108 men-patients was enrolled to the study. The mean age was 35.5+/-14.1 years. In 142 (66.7%) patients out of the total patients enrolled in the study, H. pylori was detected. H. pylori could be cultured from only 61 (43%) of them. In 16.4% of the cases, primary clarithromycin resistance was noted. After 8 weeks, seventy-seven (54.2%) of the 142 patients were reevaluated. Helicobacter pylori eradication could be achieved in 68.8% of them. The proportion of H. pylori eradication in clarithromycin-sensitive patients was 75.8% and the respective proportion was 10% for resistant cases. In the group where H. pylori was still positive the secondary resistance percentage was found to be 27.2%. CONCLUSIONS: The prevalence of primary clarithromycin resistance is relatively high in our geographical area. Secondary resistance rate was 27.2%. None of the criteria of age, gender, presence of endoscopic lesions, detected H. pylori concentration and gastritis activity showed any effect on the primary resistance. PMID- 18507128 TI - Effects of atropine sulfate and neostigmine on gastric electrical activity in human subjects--electrogastrographic study. AB - BACKGROUND/AIMS: There are few reports about the cholinergic regulation of gastric electrical activities using percutaneous electrogastrograms (EGG). To determine the effect of intravenous administration of a vagal blocker (atropine sulfate) or a vagal stimulator (neostigmine) on gastric electrical activities EGG was utilized in this study. METHODOLOGY: EGG was recorded before and after administration of a vagal blocker (atropine sulfate, 0.02mg/kg) and/or vagal stimulator (neostigmine, 0.008mg/kg) in six normal volunteers. RESULTS: After administration of atropine sulfate, the original waves on EGG almost disappearedand decreased amplitudes were detected by visual inspection in all subjects. Moreover, increase amplitude on EGG was clearly demonstrated after administration of neostigmine in all subjects. It was also shown that the effects of these medicines persisted more than 60 minutes after injection. CONCLUSIONS: These findings suggest that the EGG reflected gastric motility, and that neurological regulation of EGG was mediated through the vagal and/or cholinergic efferent pathway. PMID- 18507129 TI - Gastric emptying in patients with palliative stenting for malignant gastric outlet obstruction. AB - BACKGROUND/AIMS: Palliative stenting for gastric outlet obstruction (GOO) offers a more rapid resumption of oral intake than surgical gastrojejunostomy. Clinically, delayed gastric emptying is observed less frequently in patients with enteral stenting. The aim of this study was to conduct a functional assessment of gastric emptying after stent placement for GOO using isotope scanning. METHODOLOGY: Gastric emptying was assessed in 14 patients with GOO (4 female, 10 male; mean age 67.9 years; 8 with gastric cancer; 4 with pancreatic cancer; 1 with biliary cancer; 1 with metastasis) and 10 healthy volunteers (2 female, 8 male; mean age 31.5 years). None of the patients had undergone previous stomach surgery. The patients were studied 1 week after stent placement. Scintigraphy was performed for 2 hours following the ingestion of a labeled liquid meal. Gastric retention was evaluated at 2 hours in both groups. RESULTS: All patients underwent successful placement of stents and were able to resume an oral diet. All stents were fully deployed and no migration was seen at the time of the investigation. Retained gastric activity at 120 minutes (RGA120) was significantly greater in patients than in controls (65.4% vs. 27.5%, p=0.0128). Median survival time was 179 days in patients with T1/2 of 120 min or less and 75 days in patients with T1/2 of over 120 min. CONCLUSIONS: The results of our study show that although patients with GOO have resumed oral intake 1 week after stent placement, restoration of gastric emptying is often still incomplete. PMID- 18507130 TI - The preoperative placement of transjugular intrahepatic portosystemic shunt for treatment of a patient with portal hypertension and gastric cancer. AB - The case report describes the placement of a TIPSS preceding surgery of the upper gastrointestinal tract in a patient with liver cirrhosis and gastric carcinoma. Following two unsuccessful exploratory laparotomies, the decreased portal hypertension facilitated a curative resection of the tumor. PMID- 18507131 TI - [Real-time detection and processing of medical signals under windows using Lcard analog interfaces]. AB - Multipurpose modular software for an analog interface based on Lcard 761 is considered. Algorithms for pipeline processing of medical signals under Windows with dynamic control of computational resources are suggested. The software consists of user-friendly completable modifiable modules. The module hierarchy is based on object-oriented heritage principles, which make it possible to construct various real-time systems for long-term detection, processing, and imaging of multichannel medical signals. PMID- 18507132 TI - [Use of fuzzy models for predicting post-surgery complications in urological patients]. AB - Use of modern information technologies in combination with fuzzy decision logic makes it possible to generate prognostic models for high-confidence determination of the risk of post-surgery complications in urological patients. Problems of use of automated decision support systems based on fuzzy models for improving the accuracy of prediction of post-surgery complications in urological patients are discussed. PMID- 18507133 TI - [Prediction of genesis and development of skin diseases at bioactive sites using fuzzy decision rules]. AB - A decision system based on fuzzy decision rules is suggested to improve the quality of prediction and diagnosis of a number of widely spread skin diseases. The fuzzy decision rules use information on the bioactive sites and risk factors, as well as various data conventionally used in practical dermatology. PMID- 18507134 TI - [A method for creating fuzzy neural-network models using the MATLAB package for biomedical applications]. AB - Problems of MATLAB-based simulation of hybrid neural networks for data classification in biomedical applications are considered. The neural-network structure is used as a defuzzifier. Graphic interfaces for constructing fuzzy neural-network models are suggested. Methods for merging models constructed using different MATLAB packages and an algorithm for constructing a fuzzy neural network model are suggested. The algorithm includes both modules for tuning membership functions of the fuzzy system fuzzifier, and modules for tuning the parameters of the defuzzifier implemented as a two-layer neural-network structure of the perceptron type. PMID- 18507135 TI - [Generation of fuzzy network models taught on the basis of data structure for medical expert systems]. AB - Exploratory analysis of the characteristics and structure of the classes under study is suggested to be used for generation of fuzzy decision rules for medical applications. It is shown that the use of the fuzzy set theory in combination with computational experiment makes it possible to reduce the number of computational procedures while maintaining high quality of classification and prediction. PMID- 18507136 TI - [Sphygmogram analysis based on phase trajectories]. AB - Mattier's differential equation for parametric oscillation is suggested to be used as a method for simulating quasi-periodic processes in human body in terms of oscillation of the pulse wave produced by heart contraction. Obtained sphygmograms agree qualitatively with solutions of the Mattier's and Hill's equations at given shape of the exciting signal. A method of diagnosis based on assessment of the Mattier's equation solution stability from the phase trajectories is suggested. PMID- 18507137 TI - [Biomechanical analysis of the load on the intercondylar surface of the knee joint in frontal projection]. AB - The article is intended for experts on biomechanics, traumatologists, and orthopedists working in the field of the large joint pathology treatment. The method for analysis of the load on the knee joint presented in this article makes it possible to predict development of local necrosis of the joint surface caused by abnormal biomechanical axis of the lower extremity. The method also allows the necrosis center location to be determined with sufficient accuracy. Besides, the article provides a theoretical basis for preoperative design in the case of operative correction of the biomechanical axis of the leg intended to optimize the intense-deformed condition of the knee joint. PMID- 18507138 TI - [Criteria and methods for assessing the reliability of medical equipment. Part II: special requirements to reliability of medical equipment and methods for reliability improvement]. AB - Classification of medical equipment according to the failure effects is given. Methods for increasing the reliability of medical equipment are considered. The problem of organization of the technical state monitoring, maintenance and metrological support of medical equipment is considered from the viewpoint of legislative control. PMID- 18507139 TI - [Examination of behavioral reaction in hyperactive children with attention deficit syndrome using software-hardware systems Binatest and SVET]. AB - The problem of instrumental examination of children with socially important psychoneurological disorders (attention deficit syndrome and hyperactivity) is considered. PMID- 18507140 TI - [Bioimpedance means of skin condition monitoring during therapeutic and cosmetic procedures]. AB - Engineering and technological problems of bioimpedance skin surface mapping are considered. A typical design of a device based on a PIC 16F microcontroller is suggested. It includes a keyboard, LCD indicator, probing current generator with programmed frequency tuning, and units for probing current monitoring and bioimpedance measurement. The electrode matrix of the device is constructed using nanotechnology. A microcontroller-controlled multiplexor provides scanning of interelectrode impedance, which makes it possible to obtain the impedance image of the skin surface under the electrode matrix. The microcontroller controls the probing signal generator frequency and allows layer-by-layer images of skin under the electrode matrix to be obtained. This makes it possible to use reconstruction tomography methods for analysis and monitoring of the skin condition during therapeutic and cosmetic procedures. PMID- 18507141 TI - [A magnetic therapy apparatus with an adaptable electromagnetic spectrum for the treatment of prostatitis and gynecopathies]. AB - Problems of engineering and algorithm development of magnetic therapy apparatuses with pseudo-random radiation spectrum within the audio range for treatment of prostatitis and gynecopathies are considered. A typical design based on a PIC 16F microcontroller is suggested. It includes a keyboard, LCD indicator, audio amplifier, inducer, and software units. The problem of pseudo-random signal generation within the audio range is considered. A series of rectangular pulses is generated on a random-length interval on the basis of a three-component random vector. This series provides the required spectral characteristics of the therapeutic magnetic field and their adaptation to the therapeutic conditions and individual features of the patient. PMID- 18507142 TI - [On the 20th anniversary of the Scientific-Manufacturing Association "Elektron"]. AB - Elektron Scientific-Manufacturing Association is one of the leading domestic designers, manufacturers, and suppliers of medical X-ray equipment. It holds a leading position in development of digital diagnostic equipment. Out of a small enterprise, Elektron Scientific-Manufacturing Association has developed into one of the leading domestic manufacturers of X-ray equipment. The goal of this work was to provide information about Elektron Scientific-Manufacturing Association, its history, currently available products, and developmental strategies. PMID- 18507143 TI - [Extrapulmonary tuberculosis in risk-group children: detection methods and clinical characteristics]. AB - The paper analyzes the results of the 1995-to-2006 follow-ups of 36 children with extrapulmonary tuberculosis. Tuberculosis of the kidney, peripheral lymph nodes, and skeletal system was diagnosed in 16, 11, and 6 patients, respectively. Generalized tuberculosis was present in 1 child and cutaneous tuberculosis in 2. A contact with a tuberculosis patient, significant or progressive tuberculin sensitivity, and a lower BCG vaccination efficiency were the most important risk factors of the disease. The history of nonspecific renal and urinary tract diseases was a significant risk factor of nephrotuberculosis. The priority lines in the detection of extrapulmonary tuberculosis in children include planned systematic tuberculin diagnosis, a followup of persons who are in contact with patients with active tuberculosis, as well as dynamic bacteriological and histological studies of autoptic and biological materials. PMID- 18507144 TI - [Function in patients with chronic fibrocavernous tuberculosis]. AB - Vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), FEV1/VC%, PEF, MEF25, MEF50, MEF75, TLC, TGV, residual volume (RV), R(aw), R(in), R(ex), DLCO-SB, DLCO-SS, PaO2, and PaCO2 were determined in 62 patients with chronic fibrocavernous tuberculosis. Lung dysfunctions were detected in 96.8% of the patients. Changes in lung volumes and capacities were found in 90.3%, impaired bronchial patency was in 90.3%, and pulmonary gas exchange dysfunction was in 79.0%. The lung volume and capacity changes appeared as decreased VC and FVC, decreased and increased TLC, TGV, RV; impaired bronchial patency presented as decreased PEF, MEF25, MEF50, MEF75, and FEV1/VC%; and increased R(aw), R(in), R(ex); pulmonary gas exchange dysfunction manifested itself as reduced DLCO-SB, DLCO-SS, PaO2, and decreased and increased PaCO2. The magnitude of the observed functional changes ranges from slight to significant and drastic with a predominance of considerable and drastic changes in lung volumes and capacities and mild impairments of bronchial patency and pulmonary gas exchange function. PMID- 18507145 TI - [Increasing the efficiency of surgical treatment for disseminated spinal tuberculosis]. AB - The authors present the results of radical reconstructive operations in 80 patients with complicated spinal tuberculosis, by taking into account the duration (1.5-2 and 3-4 months) of preoperative antibacterial therapy. The efficiency of a radical reconstructive operations in the early stage (following 1.5-2 months) of hospital treatment involving intensive antibacterial and pathogenetic therapies is shown. Regress of spinal disorders is comparatively higher in the patients operated on in the early period than in the late one of antibacterial therapy. A year after surgery, there was recovery and permanent abatement of a specific spinal process in 72.5% of cases; the recovery of the support ability of the vertebral column was stated in 91.3% of cases. PMID- 18507146 TI - [Laser therapy in the complex treatment of prostatic tuberculosis at the sanatorium stage of rehabilitation]. AB - Seventy nine patients with prostatic tuberculosis were examined and treated at the specialized "Glukhovskaya" sanatorium. All the patients were divided into 2 groups: a study group and a control one. In the study group patients, low intensity laser radiotherapy was additionally performed in combination with etiological and pathogenetical treatments. The performed studies demonstrated that in the patients exposed to low-intensity laser radiation, there was a rapid relief of dysuric symptoms, cessation or diminishment of the signs of copulative dysfunction; positive changes in life quality indices and laboratory parameters were statistically significantly more frequently and more rapidly recorded. Laser therapy simultaneously was found to affect a few links of the pathogenesis of genitourinary tuberculosis, to have limited number of contraindications, to be well tolerated, and to be an effective method of the complex treatment of patients with prostatic tuberculosis during sanatorium rehabilitation. PMID- 18507147 TI - [Organization of antituberculous care at the municipal level. Section 8. Calculation of needs for drugs and other materials and their perpetual stock provision]. PMID- 18507148 TI - Incidence, antibiotic resistance and clonal relations of MRSA strains isolated from a Romanian university hospital. AB - Our aim was to estimate the frequency and characteristics ofmethicillin-resistant Staphylococcus aureus (MRSA) strains occurring in a Romanian teaching hospital. We retrospectively studied isolates from infected or colonized patients treated at the intensive care and surgical units during January 2004-December 2005. The antibiotic susceptibility of MRSA strains and the presence of mecA gene were determined. Consecutively occurring strains isolated through a three-month period were typed using pulsed field gel electrophoresis. A total of 423 S. aureus strains were identified, methicillin-resistance was detected in 211 (49.9%) strains. Most of them were multiresistant. One of the MRSA genotypes identified by PFGE was commonly recovered from patients treated in the intensive care unit. According to our results, MRSA strains were frequently isolated pathogens in our hospital and there is an urgent need to enhance infection control efforts. PMID- 18507149 TI - The novel arthritis-drug substance MCS-18 attenuates the antibody production in vivo. AB - Influence of the novel arthritis drug-substance MCS-18 on the antibody (Ab) production against tetanus toxoid (TT) and diphtheria toxoid (DT) antigens was tested in vivo. Possible involvement of MCS-18 in Toll-like receptor (TLR) signalling pathway was further considered. MATERIALS AND METHODS: Immunization of male CD1 mice was done with subcutaneous injection of TT emulsified in Freund's Complete (FCA) or Incomplete Adjuvant (FIA) and mixed diversly with MCS-18 and different test substances. To investigate the influence of TLR activation Pam3Cys and lipopolysaccharide (LPS) emulsified in FIA were tested in combinations with MCS-18. Antibody production was analysed in vivo by tetanus- or diphtheria-toxin neutralization test. RESULTS: Immunogenicity of TT was significantly enhanced if administered together with FCA or TLR agonists Pam3Cys or LPS emulsified in FIA. It was shown that MCS-18 attenuated strongly the production of anti-TT Ab if administered together with the Ab elicitor FCA or TLR agonists in various combinations. MCS-18 was also active via oral administration. DISCUSSION: These findings suggest that MCS-18 could be a potent, non-toxic antagonist or a down regulator of TLR signalling pathway. Investigations on further models are needed to establish ifMCS-18 may influence particularly the production of RA-specific auto-antibodies, too. PMID- 18507150 TI - Immobilization of polygalacturonase from Aspergillus niger onto activated polyethylene and its application in apple juice clarification. AB - The present work is focused on efficient immobilization of polygalacturonase on polyethylene matrix, followed by its application in apple juice clarification. Immobilization of polygalacturonase on activated polyethylene and its use in apple juice clarification was not reported so far. Aspergillus niger Van Tieghem (MTCC 3323) produced polygalacturonase when grown in modified Riviere's medium containing pectin as single carbon source by fed-batch culture. The enzyme was precipitated with ethanol and purified by gel filtration chromatography (Sephacryl S-100) and immobilized onto glutaraldehyde-activated polyethylene. The method is very simple and time saving for enzyme immobilization. Various characteristics of immobilized enzyme such as optimum reaction temperature and pH, temperature and pH stability, binding kinetics, efficiency of binding, reusability and metal ion effect on immobilized enzymes were evaluated in comparison to the free enzyme. Both the free and immobilized enzyme showed maximum activity at a temperature of 45 degrees C and pH 4.8. Maximum binding efficiency was 38%. The immobilized enzyme was reusable for 3 cycles with 50% loss of activity after the third cycle. Twenty-four U of immobilized enzyme at 45 degrees C and 1 h incubation time increased the transmittance of the apple juice by about 55% at 650 nm. The immobilized enzyme can be of industrial advantage in terms of sturdiness, availability, inertness, low price, reusability and temperature stability. PMID- 18507151 TI - In vitro activity of fluconazole and amphotericin B against Candida inconspicua clinical isolates as determined by the time-kill method. AB - Candida inconspicua is an emerging pathogen in immunocompromised patients possessing inherently decreased susceptibility to fluconazole. We determined the MICs and killing activity of fluconazole and amphotericin B against C. inconspicua clinical isolates as well as reference strain C. inconspicua ATCC 16783 for comparison. MICs were determined using the standard broth microdilution method. Killing rates were determined using time-kill methodology at 0.5-16 x MIC fluconazole and amphotericin B concentrations. Fluconazole and amphotericin B MIC values varied between 16-128 mg/l and 0.5-1 mg/l, respectively. In time kill assays fluconazole showed fungistatic effect at 1-16 x MIC concentrations against all tested strains after 24 h-incubation, but became fungicidal after 48 h at 4 16 x MIC concentrations. The time necessary to achieve fungicidal endpoint at 1 mg/l amphotericin B concentration ranged from 2 to 24 h. Our in vitro results confirm the data that fluconazole is ineffective against C. inconspicua at the fluconazole serum concentration attainable in humans. Amphotericin B due to its rapid killing activity seems to be a good alternative for the treatment of infections caused by C. inconspicua. PMID- 18507152 TI - Connatal listeriosis--a case report and the possibilities of microbiological diagnosis. AB - The third most frequent agent of perinatal bacterial meningitis is Listeria monocytogenes, in Hungary, its occurrence is, however, uncommon. This raises the possibility of diagnostical mistakes. A connatal listeriosis case validated microbiologically referred to in this report calls attention to Listeria, as a rare but relevant pathogen of neonatal infections. If clinical background suggests infection, the pathogenic role of L. monocytogenes should be taken into consideration. The etiological significance of the agent has to be verified by a competent clinical microbiology laboratory, since maternal listeriosis should be treated and the serious connatal manifestations should be prevented. Epidemiology of perinatal infection by L. monocytogenes, and its diagnostic tools especially the use of selective media are discussed. PMID- 18507153 TI - [Coding the differently directed behavioral actions by the putamen neurons of the monkey brain]. AB - Spike activity related to the problem of alternative choice of behavioral actions was recorded in the putamen of the monkey brain. The patterns of low and high activities were identified. Each neuron during different behaviour actions could generate any kind of patterns. The differences between neuronal compositions with patterns of high activity, at the left and right direction of the task, were obtained during decision making about the movement direction, and also at the end of the movement. Distinctions between neuronal compositions with patterns of low activity at this time, on the contrary, diminished. The neuronal compositions with patterns of low activity were much more before the conditioned signal, when the animal did not yet know the task, and at the end of the program when the problem was already solved. The data obtained show that the putamen units control different directions of actions by a multilevel address coding, mainly through reorganizing the neuronal compositions with patterns of different level activity. PMID- 18507154 TI - [Age-dependent characteristics of the skin peripheral blood flow oscillations by nonlinear dynamics methods in humans]. AB - Study of peripheral microhaemodynamics was carried out with laser Doppler flowmetry in healthy volunteers of different age groups. The ageing changes in the state of the skin peripheral blood flow, in the functioning of separate links and regulatory systems ofmicrovascular bed have been estimated in terms of relative entropy and fractal dimension values. The revealed significant age dependent decrease of relative entropy values in the respiratory rhythm ranges, the neurogenic and myogenic activities yielded some evidence concerning the reduction of the microcirculation system chaotic changes within these frequency ranges during the ageing. The significant increase of fractal dimension values in the ranges of cardio-rhythm and the endothelial activity in the oldest group with the mean age of 77 years indicated that the structural complexity of the oscillations in these frequency ranges increased during ageing. PMID- 18507155 TI - [Functional development of the vegetative regulation of the cardiovascular system in human ontogenesis]. AB - Using the functional tests method, it was shown that the ontogenetic course of development of the cardiovascular system vegetative regulation after the age 6 years might be conditionally divided into some periods. About the age of 13-14 years, the spectral structure of heart rate variability reached the level of adult humans but is accompanied with features of functional strengthening in regulative systems. About the age of 15-16 years, maturation of indices received in quiet state (heart rate, stroke volume, spectral and statistical parameters of heart rate variability) is reached. At the same time, the functional maturation of heart rate regulating systems was complete thereby providing adaptive response of the organism during the implementation of functional tests. Functional maturation of peripheral blood pressure and the systems of its regulation completed after 16 years. PMID- 18507156 TI - [Blood erythropoietin level in female reproductive age (can the reproductive tissue be the source of erythropoietin in blood?)]. AB - Blood erythropoietin level was investigated in females of reproductive age, who had normal peripheral red blood findings. It was determined that blood erythropoietin level was 1.5-2 times higher (20.04 +/- 2.10; 28.57 +/- 5.87 mU/ml) in examined females with increased mitotic activity and hyperplasia signs of cervical squamous epithelia compared with female having normal cervix (12.73 +/- 0.96 mU/ml). In females with chronic cervicitis, blood erythropoietin level was decreased (8.87 +/- 1.72 mU/ml). PMID- 18507157 TI - [Role of sodium-calcium exchanger in the myocardial protection against ischemia reperfusion injury]. AB - Present study was aimed at investigation into the role of sodium-calcium exchanger (NCX) in myocardial ischemia-reperfusion injury and ischemic preconditioning (IPC). Experiments were performed in vivo rat model of regional myocardial ischemia-reperfusion. It was shown that inhibition of reverse mode of NCX with selective blocker KB-R7943 at a dose of 10 mg/kg resulted in significant decrease in occurrence and severity of ischemic ventricular tachyarrhythmias. Furthermore, administration of KB-R7943 caused potentiation of the antiarrhythmic effect exerted by single episode of IPC. However, KB-R7943 exerted no effect on myocardial infarction size nor affected infarction size limitation by IPC. In conclusion, inhibition of reverse mode of NCX conferred significant antiarrhythmic effect against ischemic rhythm disorders but it was ineffective in terms of infarction size limitation. PMID- 18507158 TI - [Cytoskeletal proteins and heat shock proteins 27 under eccentric exercise of rats: effects of calcium L-type channel blockade]. AB - It is known that during eccentric exercise the calcium accumulation and degradation of cytoskeletal proteins take place. This study was aimed to investigation into the role of baseline calcium accumulation in cytoskeletal degradation and the way of calcium inflow into skeletal muscle un- der conditions of eccentric exercise. Wister rats were divided into control (C), eccentric exercise (EE) (one set of downhill treadmill running (-16 degrees) at a speed of 20 m. min(-1) for 40 min) and eccentric exercise plus nifedipine administration (EEN) groups (with a daily supplementation of 6.25 mg/kg nifedipine in drinking water during two days). After a 24-hour post-exercise dystrophin, layer integrity and desmin level in m. soleus were declined in EE and didn't change in EEN gr. vs. C (p < 0.05). HSP27 were decreased in EEN in comparison with EE gr. (p < 0.05). Titin and nebullin were not changed after exercise. It seems that calcium L-type channel blocker attenuates or reduces the contraction-induced damage to cytoskeletal proteins. PMID- 18507159 TI - [Microinjections of heat shock protein 70 kDa into the nucleus reticularis pontis oralis induce inhibition of rapid eye movement sleep in pigeons]. AB - Recently it was indicated that microinjections of heat shock proteins 70 kDa (Hsp70) into the third ventricle of brain in pigeons results in an increase in the duration of slow wave sleep and a decrease in somato-visceral indices. It is suggested that Hsp70 effect may be related to GABA(A) receptors activation in the preoptic area of the hypothalamus. However, what transmitter mechanisms of activation are related to the removal effect (in 2-3 hrs) of rapid eye movement sleep inhibition still remains poorly understood. To solve this problem in the present study, microinjections of Hsp70 into the Nucleus reticularis pontis oralis (NRPO) were done. It is well known that cholinergic neurons of the NRPO are crucial for rapid eye movement sleep generation. The data show that Hsp70 produces more early (for first two hrs) a decrease in number of episodes and total time of rapid eye movement sleep, a diminution of electroencephalogram (EEG) power spectra in the 9-14 Hz band, a decrease in contractile muscle activity and brain temperature. It is suggested that Hsp70 effects are realized due to activation of GABA(A) receptors in the NRPO and induced inhibition of cholinergic mechanisms of rapid eye movement sleep triggering. The microinjections of Hsp70 into the NRPO increase the slow wave sleep total time with long latency (for 8-12 hrs). This effect may be related to influence of Hsp70 on neurons population, which are responsible for slow wave sleep maintenance outside the NRPO. PMID- 18507160 TI - [Persistent pain responses under inflammation and corticosterone levels in juvenile rats born to adrenalectomized dams]. AB - Juvenile Wistar rats born to dams adrenalectomized 34 weeks before mating were used to study the intensity of indices of behavioural pain responses (number of flexing + shaking and duration of licking patterns) produced by inflammation in the formation test as well as plasma corticosterone levels during the pain response. It has been found that dams' adrenalectomy does not change either basal corticosterone levels or the indices of pain response intensity. During persistent pain (25 min after the formation injection), corticosterone levels were enhanced reliably (p < 0.05) in offspring of both intact and adrenalectomized dams. Females born to adrenalectomized dams showed the higher corticosterone levels (p = 0.008) as a response to persistent pain as compared to those born to sham-operated dams. No differences in pain indices in female offspring of adrenalectomized and sham dams were revealed. There were no sex differences in the indices of pain response and corticosterone. Thus, the pain evoked by inflammation in the formalin test has an activating effect on the hypothalamo-hypophyseal-adrenocortical system in 25-day-old pups, but the released corticosterone fails to reduce pain intensity in the formalin test as evidenced by data obtained in offspring of adrenalectomized dams. PMID- 18507161 TI - [Characteristics of responses of rabbit joint structures to intraarticular introduction of calcitonin]. AB - On a model of autoimmune arthritis in rabbits, reduction of inflammatory destructive process manifestation was observed in all joint tissues under the influence of the intraarticular introduction of calcitonin. Improving metabolic processes in the matrix of tissue-connecting elements of a joint, this specimen inhibits the inflammatory destruction of the subchondral bone and hyaline cartilage in conditions of experimental rheumatoid inflammation. PMID- 18507162 TI - [Effect of Na(+), K(+)-pump inhibitors on the sensory ganglia neurite growth]. AB - Inhibitors of Na(+), K(+)-pump belonging to the class of cardiac glycosides were investigated in organotypic tissue culture of dorsal root ganglia cells of 10-12 days old chicken embryos. The data obtained show that the application of cardiac glycosides (strophantin K and digoxin) in a wide range of concentrations controls the neurite growth in sensory neurons in the dose-dependent manner. It was shown, that at the concentrations of cardiac glycoside exceeding 1 x 10(-6) M the growth of neurites was totally inhibited. Our data indicate that cardiac glycoside have the down-regulation effect on the neurite growth. The data obtained indicate that the Na(+), K(+)-ATPase is involved in the control of the process of neurite growth as a signal transducer. PMID- 18507163 TI - [Morphofunctional development of the rabbits cecum in ontogenesis]. AB - Quick growing capacities of the caecum, bacterial number in its contents, and enzymal activities of caecal microbes were shown in the rabbit of a 20-day age. In addition, 45-day old rabbits reveal that the factors of caecal digestions reach their maximum, and then drop again at 60-day age, when the mass of the caecum in calculation on unit of the alive mass approaches to value typical of adult animal. Final shaping of the caecum functional capacities occurs at three month age. PMID- 18507164 TI - [The problems in physiology in works by Leonhard Euler]. PMID- 18507165 TI - [Topical problems in the provision of the Russian Federation's population with sanitary-and-epidemiological well-being]. AB - The most important political and organizational events considerably affecting the strategy and tactics of prevention of infectious diseases may be called at the Summit of Eight (Saint Petersburg) that has considered the problem of consolidation of the world community's efforts in counteracting the prevalence of infectious diseases, as well as the adopted specific decisions on this matter; development of a concept of financial provision of Russian initiatives in the control of infectious diseases and its realization by the Government of the Russian Federation; implementation of the national priority health project on the additional mass immunization against rubella, influenza, poliomyelitis, prevention and treatment of HIV/AIDS and viral hepatitis B and C; implementation of regional and federal programs on the population's sanitary-and-epidemiological well-being; further improvement and development of a legal and standard basis in the control of infectious diseases; reorganization of a state sanitary-and epidemiological service on a large scale; conferment of the status of a poliomyelitis-free country and a WHO certificate to the Russian Federation; development and implementation of the national action plan to maintain the achieved results; implementation of the national program in eradicating measles in the Russian Federation; implementation of complex measures against the importation and spread of revived particularly dangerous infections, such as severe acute respiratory syndrome and avian influenza and preparation of the population for a possible influenza pandemic; establishment of an executive headquarters to coordinate activities against the prevalence of avian influenza in the Russian Federation; making decisions, with the results of a meeting of the Russian Federation's State Council being kept in mind, on the intensification and coordination of activities aimed at counteracting the epidemic of HIV/AIDS in the Russian Federation, by setting up a governmental commission; involvement in the solution of biological safety problems in accordance with the "Foundations of public chemical and biological safety policy of the Russian Federation for the period till 2000 and further prospects" approved by the President of the Russian Federation; implementation of measures for sanitary area control within the framework of the goal-oriented federal program "The State Boundary of the Russian Federation" (2003-2010). PMID- 18507166 TI - [Transformation of surfactants in different ways of water disinfection]. AB - Evaluation of the efficacy ofoxidants on the destruction of bisquartemary ammonium salts (BAS) in water demonstrated that the highest destruction effectiveness was achieved upon exposure to ozone (87%), potassium permanganate (68.2%), ultraviolet radiation (37.8%), by chlorination with gas chlorine (35.4%), lime chloride (24.1%), the least destruction was achieved by chlorination with sodium hypochlorite (21.9%). There was a more than 2-fold increase in the level of low-molecular impurities upon chlorine exposure of the water containing BAS as a result of transformation processes. In ozone treatment of BAS, unlike its chlorination, there was as high as 2-fold decrease in the concentration of C6-C9 alcohols present in their composition, giving rise to C6 C9 aldehydes (0.5%) and C6-C9 organic acids (as high as 3% of the mass content). PMID- 18507167 TI - [Significance of indicator microorganisms in the assessment of a microbial risk in the occurrence of epidemic hazard in drinking water use]. AB - Summary. The paper provides comparative characteristics of water quality in the assessment of a risk for intestinal infections in drinking water use. It has shown that of the greatest predictive value is direct detection of potentially pathogenic microorganisms, as well as the integral indicator determined by glucose fermentation, such as glucose-positive coliform bacteria. Estimation of the per cent of nonstandard samples of water before its entering the distribution network and in the latter, including glucose-positive Escherichia coli GPEC, is recommended. The samples containing GPEC in a quantity of more than 2 CFU/100 ml should be singly taken into account. PMID- 18507168 TI - [Comparative assessment of radiation and chemical risks for cancer in the areas in vicinity of an atomic power station]. AB - The estimated cancer risks due to radioactive and chemical factors are assessed and compared. Their possible contribution to malignancy mortality in the population living at the areas in the vicinity of an operating atomic power station is also estimated. PMID- 18507169 TI - [The health status of children from industrial towns due ambient air pollution]. AB - The author's observations suggest that hygienic monitoring in an industrial city should be made in two areas: 1) ambient air quality and 2) human health. Ambient air quality should be monitored in each town in accordance with an individual program, by taking into account the volume and nature of hazardous substances from the stationary stations, as well as weather conditions, the planning system of residential areas, and the layout of an industrial zone. Monitoring of the population's health in the industrial town should be adapted to the forms and conditions of ambient air quality monitoring in order to reveal environmental pollution-induced changes. PMID- 18507170 TI - [Zoning of the Tyumen Region by the risk factors of water-borne intestinal infections]. PMID- 18507171 TI - [Dysbacterioses as a hygienic problem]. PMID- 18507172 TI - [Evaluation of the population's health: medicohygienic aspects]. AB - Fundamental medical and hygienic principles should be used to stratify the health status of Russia's population. Human health should be ranked, by taking into account both contributors, lifestyle, and gender, age groups, ethnicity, occupation, etc. For grading human beings, a classification of the human health status has been developed depending on acting harmful and hazardous factors. The classification involves 7 health classes: normal, adapted, borderline, pathological nonspecific (predominantly systemic chronic diseases with temporary disability), pathological specific (occupational and endemic diseases, occupational accidents, etc. with temporary disability), permanent disability (first-to-third degrees), and prefatal conditions. Class health ranking may be used for comparative assessment, and making managerial decisions on health protection of various population groups. PMID- 18507173 TI - [Impact of working conditions at a general mechanical rubber goods plant on dental morbidity in its workers]. AB - The impact of working conditions on dental morbidity was studied in workers from the Kazan general mechanical rubber goods (GMRG) plant. Analysis of the results of studying the prevalence of major dental diseases in the workers of the GMRG plant showed that the number of patients with caries, noncarious lesions, as well as periodontal diseases is far in excess of those in the control group. The studies, conducted in terms of the age and length of service of the examinees, corroborate the influence of adverse industrial factors on the prevalence of major dental diseases. PMID- 18507174 TI - [Estimation of motor qualities and skills in first-form pupils]. AB - Physical fitness, physical capacity, motor skill development were studied in 875 first-form pupils (470 boys and 405 girls) living in the ecologically poor and good districts of the town of Kirov. The ecologically poor factors associated with heavy traffic were found to be associated with reduced rapidity in boys and rapidity, flexibility, coordination capacities, and aerobic performance in girls. These factors also lowered the formation of motor skills, including the skills of light steps, walking, running, and ball figure-of-eight performance around the legs. In addition, the formation of skipping skills was decreased in girls. PMID- 18507175 TI - [Somatic trends in Moscow children]. AB - Somatic trends was considered in Moscow children in a wide age range of 3 to 17 years, by attracting several series of data since the 1990s. To have the greater informative value in data analyses, investigators use the normalized rather than absolute values of various anthropometric signs in the considered data series versus the 2005 data in the age range of 3 to 7 years and versus the 1980s data in the range of 8 to 17 years. The overall somatic tendency in Moscow children aged 3 to 17 years in past decades is towards a transverse body development and some dolichomorphism with a continuous increase in longitudinal skeletal sizes. These processes are most pronounced in girls than in boys and preschool children than in school ones. The stronger somatic trend in preschool children can be associated with that they are, as compared with schoolchildren, neophytes in the compact anthropogenic space and have been, in the past decades, involved in the neurogenic growth stimulators world (a computer and computer games, unlimited TV seeing, inevitable intensive preparation for new-generation schools--increased intellectual-load lyceums and colleges). The decreased transverse body development and overall constitution asthenization seem to result from somatic muscular component abatement with no clear trends in adipopexis in schoolchildren and with an increase in subcutaneous fat deposition in preschool children. PMID- 18507176 TI - [Impact of atmospheric pollution on preschool children's health]. AB - The implication of relatively low atmospheric pollution on preschool children's health was studied. There were no significant differences in parameters, such as health index, the frequency of functional impairments, acute morbidity rates in children living in the areas of concentration of low-power air pollution sources and in the pure areas. However, a significantly larger number of ill children and children with Staphylococcus aureus being detected on the nasal mucosa were identified in the relatively high atmospheric air pollution areas. PMID- 18507177 TI - [Adolescents' physical development in a poor environmental area]. AB - Physical and sexual developments were determined in 285 schoolchildren aged 12 to 17 years from the town of Karabash, an area of ecological ill-being in the environmental levels of heavy metal salts. The atmospheric air pollution was in parallel monitored at route observational stations. It was found that 39% of the schoolchildren had disharmonic physical development; of them a fourth had lower body weight and decreased chest circumference with normal growth indices. The girls were observed to have a high frequency of a microsomatotype of harmonic physical development and disharmonic physical development mainly at the expense of underweight (52.9% of all the disharmonically developed girls) as compared with the boys. There was a preponderance of female adolescents with sexual retardation. In the population of Karabash girls, the mean age at menarche was 12 years and 9 months +/- 1.5 months. PMID- 18507178 TI - [Visual reactions in children and adolescents on reading editions of various formats]. AB - At present, 93-96% of the respondent schoolchildren read extraschool literature. An investigation of the impact of edition format on readability has ascertained that due to the uniform distribution of brightness in the field of vision, large newspaper fonts prevents the development of visual fatigue and provides significantly higher performance and quality of visual work as compared with low book fonts. Experimental studies have yielded data on the stability of a near vision point; this phenomenon may be considered as adaptation to a learning process and myopia developing during learning may be regarded as the price of this adaptation. PMID- 18507179 TI - [Cryomassage and phoresis of pharmaceutical substances in therapy of patients with degenerative spine disease after decompression operations on the lumbosacral level]. PMID- 18507180 TI - [Use of ozonotherapy (OT) in combined treatment of patients with discirculatory encephalopathy (DEP)]. AB - The purpose of the work was assessment of clinical efficiency of OT and study of cerebral blood flow indices in combined treatment of DEP patients, including OT. It was detected that addition of intravenous OT in a complex of planned therapy of DEP patients at the age about 60 years leaded to more significant and more rapid (vs. control groups) regress of DEP symptoms, in the first place headache, ear noise, sleeplessness, memory reduce. Decrease in objective manifestation of neurological syndromes such as dysmnesic, akineticorigid, asthenic ones. Blood lipid spectrum, blood glucose level, blood coagulant system and cerebral blood flow indices were improved. These effects were more expressive in patients with the age less than 60 years. In control subgroups these changes were less expressed. Thus, of OT in combined treatment of patients with DEP improved efficiency of drug therapy and accelerated regress of various symptoms of this pathology, especially in patients with the age less than 60 years. PMID- 18507181 TI - [Efficiency of short courses of physiotherapy with the use of energy metabolism regulators in patients with dorsopathy]. AB - The assessment of procedure tolerance, their efficiency at shortening of courses of dorsopathies physiotherapy to 12-14 days made possible to detect presence of unfavorable reactions to treatment in 32% of cases. Regulator of energy metabolism, included in the therapy, assists to good tolerance to shortened courses of dorsopathy rehabilitation (frequency of presence and expression degree of reaction to treatment reduced to 15%), stabilization of energy system in patients' organism, absence of hyper activation and stress of adaptive mechanisms, maintenance of compensatory abilities reserves. PMID- 18507182 TI - [Features of dynamic magnetotherapy effect on microcirculation in arterial hypertension]. PMID- 18507183 TI - [Sylvinite speleochamber and general chloride sodium baths in medical rehabilitation of patients with chronic obstructive pulmonary disease]. AB - Clinico-laboratory, functional and bronchoscopic examinations were performed in 120 patients with chronic obstructive pulmonary disease. 50 patients (41.7%) were in stage of low-intensity exacerbation, 70 patients (58.3%)--in remission stage. 55 patients (45.8%) had I degree respiratory failure, 65 patients (54.2%)--II degree respiratory failure. Easy degree of disease was detected in 58 patients (48.3%), mid degree--in 62 patients (51.7%). Possibility and reasonability of the use of artificial sylvinite speleotherapy and general chloride sodium baths in patients with chronic obstructive pulmonary disease were proved. Clinical results were confirmed by data of blood morphology and biochemical analysis, microscopy of sputum, spirometry, bronchoscopy, dynamics of immunological indices. PMID- 18507184 TI - [Effect of physical rehabilitation on endothelial dysfunction in chronic inflammatory joint diseases]. AB - The purpose of the study was to determine the degree of influence of physical exercises on endothelial dysfunction in patients with chronic articulation inflammatory diseases on the example of rheumatoid arthritis (RA) and osteoarthritis (OA). 41 patients with chronic articulation inflammatory diseases were examined. Endothelial dysfunction was detected in 37% OA patients and 77% RA patients. The results of the study show significantly higher degree of influence of physical exercises on endothelial dysfunction in comparison with drug therapy. PMID- 18507185 TI - [Clinico-physiological rationale for the use of the mesodiencephalic modulation in combination with synobiotics in the irritable bowel syndrome]. PMID- 18507186 TI - [Thalassotherapy as a method of physioprophylaxis of late gestosis]. PMID- 18507187 TI - [Spa treatment as the major stage for patients with metabolic syndrome rehabilitation]. PMID- 18507188 TI - [Active orthotest as a marker of adequacy of rehabilitation in patients with myocardial infarction]. PMID- 18507190 TI - [Therapeutic and recreational potential of the rest home "Frontier Guard of Russia" of the Federal Security Service of the Russian Federation]. PMID- 18507189 TI - [Training with biological feedback in sportsmen]. PMID- 18507191 TI - [Motion activity module--a new physical exercise technology in the treatment and prophylaxis of spine osteochondrosis]. PMID- 18507192 TI - Tuberculosis, one disease, many faces. PMID- 18507193 TI - Observational case-control study of non-invasive ventilation in patients with ARDS. AB - BACKGROUND: The application of non-invasive pressure support ventilation (NIPSV) in patients with acute lung injury or ARDS remains controversial despite recent promising results. Data in rather homogeneous ARDS groups is lacking. OBJECTIVE: To compare the outcome of NIPSV-treated patients satisfying the diagnostic criteria for primary (pulmonary) ARDS (ARDSp) and presenting without distant organ failures at admission, with those of a matched control group treated in the same ICU with endotracheal mechanical ventilation (ETMV). METHODS: We applied NIPSV in 12 immunocompetent and collaborative patients who met the above cited criteria. NIPSV failure rate, short-term oxygenation, length of stay, mortality rate and complications were analyzed and compared with a control group of 12 intubated ARDSp-patients matched for age, SAPS II, PaO2/FiO2 and pH at admission. RESULTS: NIPSV failed in 4 patients developing distant organ failures. Compared to the ETMV control group, NIPSV success patients had reduced cumulative time on ventilation (p = 0.001) and length of ICU stay (p = 0.004). After the first 60' of ventilation, oxygenation improved more in the NIPSV than in the ETMV group (146 +/- 52 mmHg vs 109 +/- 34 mmHg; p = 0.05). The overall ICU mortality rate did not differ significantly between the groups but tended to be higher in the NIPSV group. CONCLUSIONS: In ARDSp patients without distant organ failures at admission and during the disease course, NIPSV might be a suitable alternative to invasive ventilation; however, the real effects on outcome of NIPSV applied to stable homogeneous subgroups of ARDS patients merit further investigations in randomised studies. PMID- 18507194 TI - Heart and lungs in COPD. Close friends in real life--separate in daily medical practice? AB - Chronic obstructive pulmonary disease (COPD) is associated with a 2-3 times higher rate of cardiovascular disorders (CVD) which is independent of other risk factors. A low FEV1 is a specific predictor of mortality as a result of cardiac causes, even stronger than increased cholesterol: for each 10% reduction of FEV1, cardiovascular mortality increases by 28%. The main causes of death among COPD patients are of cardiovascular origin. COPD and CVD have two major risk factors in common - advanced age and tobacco smoking. The search for a pathogenetic link between the two conditions focuses mainly on systemic extension of pulmonary inflammation. Despite such a frequent association, pulmonologists and cardiologists in both the clinical and the research settings often underestimate the importance of a correct diagnosis and severity stratification of the two combined conditions. Spirometry, in particular, is largely underprescribed. Missed diagnosis and severity stratification, incomplete knowledge of adverse drug events and lack of resources lead to undertreatment of patients combining COPD and CVD, and in particular, the underuse of beta-blockers, inhaled bronchodilators and rehabilitation. Clinical studies focusing on this group of patients should be promoted in the future to test therapies and manage options. Furthermore, efforts must be made to improve the present standards of care, which falls short of recommended levels, starting from the often-neglected use of spirometry to confirm a diagnosis of COPD. PMID- 18507195 TI - Could IFN-gamma predict the development of residual pleural thickening in tuberculous pleurisy? AB - BACKGROUND: The aim of our study was to identify predictive factors for the development of residual pleural thickening (RPT) in patients with tuberculous pleurisy (TP). METHODS: A retrospective study of patients with pleural tuberculosis. The clinical and radiological characteristics, and measurements of microbiological and biochemical parameters or markers such as adenosine deaminase (ADA), interferon-gamma (IFN-gamma) and vascular endothelial growth factor (VEGF) in pleural fluid were studied. RESULTS: Thirty one patients (24 male and 7 female) with a mean age of 55.9 years were studied. There were 25 (80.6%) patients with RPT > 2 mm and 6 (19.4%) patients without RPT. Ten patients (32.2%) had RPT > or = 10 mm. The rate of pleural thickening was less in small effusions (p<0.05). IFN-gamma was higher in patients with RPT > or = 10 mm (p < 0.05) in comparison with those with RPT < 10 mm. CONCLUSIONS: Pleural fluid IFN-gamma may deserve further investigation in order to build up preventive and therapeutic strategies against RPT and its clinical complications. PMID- 18507196 TI - Human genes in TB infection: their role in immune response. AB - Tuberculosis (TB) caused by the human pathogen Mycobacterium tuberculosis, is the leading cause of morbidity and mortality caused by infectious agents worldwide. Recently, there has been an ongoing concern about the clarification of the role of specific human genes and their polymorphisms involved in TB infection. In the vast majority of individuals, innate immune pathways and T-helper 1 (Th1) cell mediated immunity are activated resulting in the lysis of the bacterium. Firstly, PTPN22 R620W polymorphism is involved in the response to cases of infection. The Arg753Gln polymorphism in TLR-2 leads to a weaker response against the M. tuberculosis. The gene of the vitamin D receptor (VDR) has a few polymorphisms (BsmI, ApaI, Taq1, FokI) whose mixed genotypes alter the immune response. Solute carrier family 11 member (SLC11A1) is a proton/divalent cation antiporter that is more familiar by its former name NRAMP1 (natural resistance associated macrophage protein 1) and can affect M. tuberculosis growth. Polymorphisms of cytokines such as IL-10, IL-6, IFN-g, TNF-a, TGF-b1 can affect the immune response in various ways. Finally, a major role is played by M. tuberculosis antigens and the Ras associated small GTP-ase 33A. As far as we know this is the first review that collates all these polymorphisms in order to give a comprehensive image of the field, which is currently evolving. PMID- 18507197 TI - An unusual association between Mycobacterium tuberculosis and Aspergillus fumigatus. AB - Allergic bronchopulmonary aspergillosis (ABPA) is rarely described outside the setting of asthma or cystic fibrosis. The occurrence of ABPA in other structural lung diseases included scars of old healed pulmonary tuberculosis (PTB) is also unknown. In this case, we report a 62-year old lady treated for PTB 40 years ago who presented with increasing dyspnea on exertion, cough with expectoration of blackish brown mucus plugs and wheezing. High-resolution computed tomographic scan of the thorax showed parenchymal fibrosis and volume loss in left upper lobe while central bronchiectasis, mosaic attenuation, centrilobular nodules with a tree-in-bud pattern were observed in the other lobes. Investigations revealed a diagnosis of ABPA. The patient was treated with prednisolone and showed a significant response. We review the current literature on this unusual association of previous and cured TB with ABPA, and also discuss the hypothesis of this possible relationship. PMID- 18507198 TI - Two cases of asbestosis and one case of rounded atelectasis due to non occupational asbestos exposure. AB - Asbestos is a well-known cause of several neoplastic (malignant mesothelioma, lung cancer) and non-neoplastic (asbestosis, pleuropathies) occupational diseases. Lower-level exposure in the general environment may induce pleural plaques and thickenings, and is associated with an increased mesothelioma risk. We present two patients (a 68-year-old man and a 72-year-old woman) who developed asbestosis (in association with pleural plaques and calcifications), and a 78 year-old man who developed rounded atelectasis (with pleural plaques and benign effusion), after living for several decades in the proximity of large Italian asbestos-cement plant. None of them had been exposed to asbestos occupationally. Besides living in a contaminated area, the woman used to clean the work clothes of her brother, who was employed in the local asbestos factory. The three cases indicate that non-neoplastic, long-latency asbestos-related diseases which are usually observed as a consequence of occupational exposures, may rarely develop in subjects living in contaminated geographical sites and buildings. These unusual environmental diseases raise the diagnostic problem of differentiating them from other, more common respiratory illnesses, and impose the duties of patient notification, assessment and follow-up, to assess the possibility of progression of disease and increased neoplastic risk. PMID- 18507199 TI - Relapsing bronchiolitis obliterans organising pneumonia and chronic sarcoidosis in an atopic asthmatic patient. AB - Asthma is thought to be a Th2 disease while sarcoidosis is considered a Th1 granulomatous disorder. Organising pneumonia is a histologic pattern of lung injury. When it has no recognisable cause it is defined as cryptogenic organising pneumonia. We herein report the case of a patient with recurrent and steroid sensitive organising pneumonia associated with chronic sarcoidosis in an atopic, moderate persistent asthmatic patient. Each disease has been documented with transbronchial biopsies and recurrence of organising pneumonia was suggested by clinical features and by follow up HRCT which shows distinctive signs even in associated disease. Steroids are the mainstay of therapy for these disorders and especially for the consolidated processes typical of organising pneumonia but prognostic indices for relapse and progression are lacking. PMID- 18507200 TI - Breast cancer epidemiology in blacks and whites: disparities in incidence, mortality, survival rates and histology. AB - BACKGROUND: This study presents black-white breast cancer statistics, tumor histology and receptor status, and treatment patterns for all ages and by age groups (< 40, 40-49, and > or = 50). METHODS: The study used data from the National Cancer Institute (NCI) Surveillance, Epidemiology and End Results (SEER) program for the time period 1995-2004. Age-adjusted incidence, mortality, relative survival rates, tumor grade, histology and receptor status, and treatment patterns for invasive breast cancer were calculated for nine SEER cancer registries for 1995-2004. RESULTS: Invasive breast cancer age-adjusted incidence for black women age < 40 was significantly higher than those for white women (rate ratio = 1.16, 95% confidence interval: 1.10-1.23). Age-adjusted mortality rate for black women age < 40 was twice that for white women. Compared to white women, black women were significantly more likely to be diagnosed with regional or distant disease, have lower relative five-year survival rate and have higher likelihood of being diagnosed with tumors with poorer prognosis. Black women were less likely to receive breast cancer surgery as part of the treatment plan. CONCLUSIONS: Race/ethnic disparities in invasive breast cancer epidemiology, prognostic indicators and treatment patterns exist between black and white women. The study findings support the need for innovative research, especially on the multifaceted determinants of the differential epidemiology of breast cancer. Equally important, there is a need for evidence-guided equal delivery of quality care to eliminate breast cancer disparities among black women. PMID- 18507201 TI - Survival difference between non-Hispanic black and non-Hispanic white women with localized breast cancer: the impact of guideline-concordant therapy. AB - OBJECTIVES: This study examined the impact of guideline-concordant therapy on the survival difference between non-Hispanic black (NHB) and non-Hispanic white (NHW) women with localized breast cancer. METHODS: Data analyzed were from the CDC's NPCR Patterns of Care study in which seven population-based state cancer registries participated. We randomly selected 2,362 women who were diagnosed with a first primary localized breast cancer in 1997. Data were abstracted from hospital records, supplemented by information from physician offices and by linkages with state vital records and the National Death Index database. RESULTS: NHB women were more likely than NHW women to receive breast conserving surgery without radiation therapy. In addition, the percentage of NHB women with hormone receptor-positive tumors who received hormonal therapy was lower than that of NHW women. Among those with a tumor size > 3 cm, NHB women were more likely than NHW women to receive multiagent chemotherapy. After controlling for age, the risk of dying from all causes of death was 2.35 times as high for NHB women compared to NHW women. Controlling for treatment further reduced black-white difference in survival with adjustment for sociodemographic and clinical variables. CONCLUSION: NHB women were less likely than NHW women to receive guideline-concordant radiation therapy after breast conserving therapy and hormonal therapy but were more likely to receive chemotherapy. Racial differences in treatment contribute significantly to the worse survival of NHB women compared with NHW women. PMID- 18507202 TI - A pilot study to explore the usefulness of antibody array in childhood atopic dermatitis. AB - BACKGROUND: The pathophysiology of childhood atopic dermatitis (AD) involves complex interactions among cellular, humoral, cytokine and chemokine systems. OBJECTIVE: To evaluate protein expressions using antibody microarray. METHODS: Severity-nine proteins were assayed using antibody microarray on AD patients age < 18 years. Disease severity was assessed with the SCORing Atopic Dermatitis (SCORAD) and Nottingham Eczema Severity Score (NESS), and quality of life with the Children Dermatology Life Quality Index (CDLQI). Serum IgE levels were also assessed. Normal subjects without atopy were used as controls. Cytokines, chemokines and a wide array of proteins were assayed with RayBio Human Cytokine Antibody Array V (RayBiotech, Norcross, GA). RESULTS: Nine Chinese children with AD and four normal subjects were recruited. The median SCORAD was 60.7. Among the 79 proteins, the levels of BDNF, Fit-3 ligand, IL-8, IL-16, LIGHT, MIP-1beta, MIP 3alpha, NAP-2, PARC, TGF-beta2 and TIMP-2 were significantly different from the controls. Nevertheless, no significance was found when adjusted for multiple comparisons using p = 0.0006. Some of these markers showed significant correlations with various components of SCORAD, NESS and CDLQI. The serum IgE level as a marker of atopy correlates significantly with BDNF, LIGHT, PARC and TIMP-2. CONCLUSIONS: The serum levels of BDNF, LIGHT, PARC and TIMP-2 correlate to IgE as a marker of atopy. Although targeting chemokines and chemokine receptors may offer new opportunities for therapeutic interventions in AD, protein assay with cytokine antibody array was generally not helpful in identifying specific molecules pertinent to AD activity. PMID- 18507203 TI - Family matters in mammography screening among African-American women age > 40. AB - PURPOSE: To examine how family history of cancer influences the mammography screening behaviors of asymptomatic African-American women. METHODS: Using the National Health Interview Survey's 2000 Cancer Control Module, the authors performed bivariate analyses and multivariate logistic regressions with SAS/SUDAAN due to the complex sampling design. RESULTS: Of the 1,531 African American women in the final sample, 38% had a family history of cancer. Women with a family history of cancer were 39% more likely to have a recent mammogram compared to women with no family history of cancer (OR = 1.39; 95% CI: 1.06-1.81; p < 0.05). Eighty-five percent of African-American women aged > 40 with a family history of cancer indicated having a mammogram in the past compared to nearly 70% of African-American women without a family history of cancer. CONCLUSION: Family history of any cancer independently and positively predicted mammography screening behaviors among asymptomatic African-American women. This suggests that African-American women with a history of cancer in their family are more likely (and perhaps more motivated) to engage in early cancer detection practices. PMID- 18507204 TI - Characteristics of breast cancer patients with central nervous system metastases: a single-center experience. AB - The aim of this study was to assess the characteristics of breast cancer patients with central nervous system (CNS) metastases and factors associated with survival after development of CNS metastasis. One-hundred-forty-four patients with brain metastases were retrospectively analyzed. Median age at the time of brain metastasis diagnosis was 48.9. Median time between initial diagnosis and development of brain metastasis was 36 months. Fourteen cases had leptomeningeal involvement. Twenty-two patients (15.3%) had single metastasis. Ten percent of the patients had surgery, 94% had radiotherapy and 63% had chemotherapy. Median survival after development of brain metastasis was 7.4 months. Survival of patients with single metastasis was significantly longer than those with multiple metastases (33.5 vs. 6.5 months, p = 0.0006). Survival of patients who received chemotherapy was significantly longer than those who received radiotherapy alone (9.9 vs. 2 months, p < 0.0001). In multivariate Cox regression analyses, presence of single metastasis and application of chemotherapy were the only significant factors associated with better survival (p = 0.047 and p < 0.0001, respectively). Age at initial diagnosis or at the time of brain metastasis, time from initial diagnosis to development of brain metastasis, menopausal status, tumor stage, grade, hormone receptor or HER2 status individually were not associated with survival. In this study, survival after the diagnosis of CNS metastases appeared to be affected by patient characteristics rather than biologic characteristics of the tumor. This is probably secondary to the lack of effective treatment options in these patients and overall poor prognosis. PMID- 18507205 TI - Predictors and barriers to timely medical follow-up after cardiovascular disease risk factor screening according to race/ethnicity. AB - BACKGROUND: The purpose of this study was to assess followup practices among individuals found to have elevated cardiovascular disease (CVD) risk factors in a screening and educational outreach. METHODS: Participants in the National Heart, Lung and Blood Institute Family Intervention Trial for Heart Health (FIT Heart) who were found to have hypertension, prehypertension, suboptimal lipids and/or abnormal blood glucose were included in this study (N = 214, mean age 49 +/- 13, 64% female, 33% nonwhite). Contact was made at two weeks, six weeks and three months to determine if medical follow-up was initiated. Barriers to nonadherence were assessed. RESULTS: After two weeks, significantly more whites had medical follow up compared to nonwhites (34% vs. 20%, p = 0.04). Racial/ethnic minorities were more likely to report that not having a doctor was a barrier (30% vs. 11%, p = 0.02). Non-whites were more likely to return to the study staff for followup rather than an outside physician (32% vs. 15%, p = 0.001). CONCLUSION: Racial/ethnic minorities with elevated CVD risk factors may have delayed medical follow-up compared to whites, and this may be attributable to lack of access to a doctor. These data suggest that improving access to care may reduce racial/ethnic disparities in risk factor management and CVD outcomes. PMID- 18507206 TI - The relationship between racial identity, income, stress and C-reactive protein among parous women: implications for preterm birth disparity research. AB - The persistent racial disparity in preterm birth (PTB)remains one of the most obvious yet poorly understood health disparities in the United States, and current evidence suggests that maternal stress, infection and inflammation may play an important role in the etiology of PTB. In this context, we assessed the complex relationships among racial identity; socioeconomic status (SES); psychosocial factors; and serum C-reactive protein (CRP), an inflammatory biomarker, among parous women in King County, WA. African-American women consistently reported a higher number of stressful life events than white American women (4.6 vs. 2.9, p < 0.001), as well as slightly higher levels of perceived stress and lower social support (24.7 vs. 22.2, p = 0.011, and 3.4 vs. 3.6, p = 0.06, respectively). In the multivariate analysis, African-American race, low-income status and their interaction were all independently associated with CRP; when further adjusted for proximal psychosocial, behavioral and infectious factors, race and income associations were significantly reduced. Stressful life events score was the single best proximal predictor of CRP levels (beta = 0.07 per event,p < 0.001), while perceived stress and social support were not significantly related to CRP. These results support the hypothesis that differences in CRP by racial identity and income may be mediated by differences in proximal risk factors, including stressful life events and health behaviors such as smoking. Objective life event stressors may be important to consider in future studies investigating a potential inflammatory etiology for preterm birth. PMID- 18507207 TI - The impact of nurse case management home visitation on birth outcomes in African American women. AB - The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Uniformed Services University of the Health Sciences, Department of the Navy, Department of Defense or the U. S. government. Dr. Wells is a military service member (employee of the U.S. government). This work was prepared as a part of her official duties. Title 17, USC Section 101 defines a U.S. government work as a work prepared by a military service member or employee of the U.S. government as part of the person's official duties. Despite substantial reductions in U.S. infant mortality rates, racial disparities persist, with black Americans experiencing 2.4 times the rate of their white counterparts. Low birthweight and preterm delivery contribute to this disparity. METHODS: To examine the association between antepartum nurse case management home visitation and the occurrence of low birthweight and preterm deliveries in African-American women in Montgomery County, MD, a retrospective cohort study was conducted using existing data from 109 mothers who were enrolled in the Black Babies Start More Infants Living Equally Healthy (SMILE) program. Logistic regression analysis was used. RESULTS: Women who received antepartum home visits were 0.37 (CI 0.15-0.94) times less likely to experience preterm delivery than women who did not receive antepartum home visits. The effect of antepartum home visits on preterm delivery was independent of level of prenatal care, negative life events and number of prior live births. There was no significant association between antepartum home visits and low birthweight. CONCLUSION: Antepartum home visits appeared to be protective against preterm delivery and could contribute to reducing racial disparities in infant mortality. Further study is needed to understand and replicate specific program components that may contribute to improved birth outcomes in African American women. PMID- 18507208 TI - Racial/ethnic sexual health disparities among incarcerated women. AB - Rates of sexually transmitted infections (STIs) in women in U.S. corrections facilities are higher than rates in community samples. Research that combines behavioral correlates of STI with STI history by race/ethnicity has not been done in incarcerated women. The purpose of this study was to compare by race/ethnicity self-reported sexual risk behaviors with self-reported history of STI in an incarcerated sample. An interviewer administered a questionnaire to 428 incarcerated women. Blacks were more likely to report consistent condom use in the three months prior to incarceration (47% vs. 28%, p < 0.05), and Hispanics were less likely to report sex work than were whites (16% vs. 39%, p < 0.05). Whites were more likely than blacks to report having had an unplanned pregnancy (88% vs. 67%, p < 0.05). Despite having lower self-reported risk on several measures, Blacks were more likely to report history of STI (65% vs. 40%, p < 0.05). The correctional setting is an opportune place to better understand and address the complex issue of sexual health disparities. PMID- 18507209 TI - Disparities in the utilization of high-volume hospitals for total knee replacement. AB - OBJECTIVE: Higher hospital surgical volumes have been associated with lower complication rates following total knee replacement. The objective of this study is to identify the characteristics of patients who undergo total knee replacement at high-volume hospitals and their differences from those who receive care at low volume hospitals. METHODS: Discharge data from patients undergoing total knee replacement in California from 1991-2001 were analyzed. Hospitals were classified into three tiers of low, intermediate or high surgical volume. The relationship between race/ethnicity and insurance status and the utilization of low-volume and high-volume hospitals were examined by creating separate logistic regression models that corrected for covariates, including age, gender and comorbidity. RESULTS: This study analyzed 222,684 primary total knee replacements during the study period. Patients of non-Caucasian race/ethnicity had higher relative risk ratios for being treated at a low-volume center, including black patients [relative risk ratio (RRR) = 1.73, 95% confidence Interval (CI): 1.09-2.76, p = 0.02]; Hispanic patients (RRR = 3.13, 95% CI: 2.31-4.23, p < 0.001) and Asian/Pacific Islanders (RRR = 2.95, 95% CI: 1.89-4.62, p < 0.001). Medicaid insurance was also an independent predictor of treatment at low-volume hospitals. Age and comorbidity were not statistically significant predictors for receiving care at low-volume centers. CONCLUSIONS: There are substantial disparities in the characteristics of patients receiving care at hospitals performing a high volume of total knee replacements. Black, Hispanic and Asian race/ethnicity as well as Medicaid insurance were predictors of utilization of a low-volume hospital. This study supports the need to consider racial and socioeconomic disparities in efforts to improve the quality of care of patients undergoing total knee replacement at lower-volume hospitals. PMID- 18507210 TI - The relative risk of cardiovascular death among racial and ethnic minorities with metabolic syndrome: data from the NHANES-II mortality follow-up. AB - The tendency for selected cardiovascular disease (CVD) risk factors to occur in clusters has led to the description of metabolic syndrome (MetS). The relative impact of the individual risk factor on the overall relative risk (RR) for cardiovascular death from metabolic syndrome is not well established and may differ across the different racial/ethnic groups. Using data from the National Health and Nutrition Examination Survey (NHANES II) mortality follow-up (NH2MS), we determined the prevalence and RR of cardiovascular death for individual components in the overall population and across racial and ethnic groups. The prevalence of MetS components varied significantly across gender and racial/ethnic groupings. The RR for CVD also varies for the number and different components of MetS. The adjusted RR for cardiovascular death was highest with diabetes (3.23; 95% CI: 2.70-3.88), elevated blood pressure (2.28; 95% CI: 1.94 2.67) and high triglycerides (1.63; 95% CI: 1.34-2.00). Although the RR for cardiovascular death differs significantly for some of the different components, the overall findings were similar across racial/ethnic groups. The two components that confer the highest risks for death are more prevalent in African Americans. We concluded that the RR of cardiovascular death associated with the diagnosis of MetS varies depending on the number and components used to establish the diagnosis of MetS and the racial/ethnic characteristic of the participants. PMID- 18507211 TI - Lateral rectus muscle palsy, facial numbness and ataxia as the initial manifestation of multiple sclerosis. AB - Lateral rectus muscle (LRM) palsy due to a nuclear or fascicular sixth nerve lesion is rare as the presenting sign of multiple sclerosis (MS). It is more common to find this palsy in the company of other nearby cranial nerves deficits. Facial numbness in association with a LRM palsy or paresis may go unappreciated and therefore underreported. We report an unusual patient with a LRM palsy, facial anesthesia, ataxia and as the initial manifestation of MS. There was a demyelinating lesion in the pons seen on magnetic resonance imaging (MRI). PMID- 18507212 TI - University of Pennsylvania surgeon receives grant to develop "molecular cardiac surgery" as a possible alternative to heart transplant. PMID- 18507213 TI - Sleep duration among Black and White Americans. PMID- 18507214 TI - Request for retraction of published statement on EIA. PMID- 18507215 TI - Veterinary education in South Africa: the Classes of 1932 & 1933. AB - The Class of 1933 of the Onderstepoort Faculty was the smallest in its history, having only 1 student for much of the BVSc course. The photographs of the Classes of 1932 and 1933 are both of the customary composite type. Not only the graduates, but also the Dean, Prof. Dr P J du Toit, as well as the presumed Acting Dean, Prof. Dr G de Kock, and the head of the students' hostel, Mr W O Neitz, feature. Concise descriptions are given of the life histories of the 7 graduates. Once again their careers show considerable variation. Although they all initially joined Field Services as state veterinarians, none spent their entire careers in that division. Three spent virtually their entire careers at Onderstepoort, also teaching (part-time) at the Faculty. After a most impressive start to his career, a 4th graduate died at the age of only 30 shortly after leaving Onderstepoort for municipal (public health) service. A 5th spent most of his career in private practice while also serving the veterinary profession for 8 years in the high office of president of its association. One spent the greater part of his career in municipal (public health) service and another went into private practice, eventually also farming with citrus. PMID- 18507216 TI - Acute normovolaemic haemodilution--is it a solution to reduce perioperative blood transfusions? AB - Haemodilution is a technique used to reduce perioperative homologous blood transfusions. Haemodilution is a poorly investigated technique in veterinary medicine. This article reviews haemodilution as a potential technique to reduce perioperative homologous blood transfusions. The history of haemodilution is briefly reviewed followed by the mathematical basis to haemodilution. The issue of critical oxygen delivery and its implications for haemodilution are discussed. The effects of haemodilution on the patient, including the effects on oxygen transport, blood flow and coagulation are discussed as well as the use of colloids, fluids and blood components in haemodilution. The success and failure of haemodilution in human clinical trials and experimental evidence is discussed. Some guidelines are given for the use of haemodilution in small animal patients in the perioperative setting. It appears in all likelihood that haemodilution has a limited application in cats and other small patients. Haemodilution is most beneficial when the initial haematocrit is high, a low haemodiluted haematocrit is achieved, the patients circulating volume is large and a large amount of blood was lost. It is important to avoid haemoconcentration during surgery as this increases red blood cell loss. Haemodilution is not a substitute for poor surgical technique and inadequate haemostasis intra-operatively. Intravascular volume should be maintained throughout the procedure. PMID- 18507217 TI - Factors related to shell deaths during artificial incubation of ostrich eggs. AB - The ostrich industry experiences a high rate of embryonic mortalities during artificial incubation of eggs. Embryonic deaths were studied from data recorded on 37,740 fertile eggs incubated artificially during the 1998-2005 breeding seasons. Roughly 10,000 eggs that sustained embryonic mortalities were classified according to the stage and nature of death, i.e. before 21 days of incubation, after 21 days of incubation, deaths after pipping and rotten eggs. Although infection may have played a role in approximately 1300 rotten eggs, no detailed knowledge of the pathogens involved was available. The remainder of deaths could not be related to pathogens and the deaths were thus generally referred to as non infectious. The overall level of embryonic mortality in all the eggs studied was 28.5 %. Overall embryonic mortality was affected by incubator, with higher levels (57.0 %) found in eggs incubated in an African Incubator and also in eggs that were transferred between incubators during incubation (38.1%). Overall embryonic mortality also increased in eggs produced by older females. Eggs produced in the autumn had the highest level of embryonic mortality at 53.6 %, whereas eggs produced in the winter had a marginally higher level of embryonic mortalities of 29.2 % compared with eggs produced during summer (27.4 %). Eggs produced by South African (SA) Black males crossed to Zimbabwean Blue females had high levels of embryonic losses of 45.7 %. The embryonic mortality of eggs produced by SA Blacks or Zimbabwean Blue breeding birds subjected to pure breeding was similar at approximately 33-34 %, but embryonic mortality was improved in eggs produced by Zimbabwean Blue males crossed to SA Black females (27 %). Embryonic mortality was increased in eggs that were set directly (32.0 %) or subjected to longer than 6 days of storage (43.5 %). Embryonic mortality was affected by year. The results that were obtained will assist in determining non-infectious factors that have a negative effect on hatching success. Steps can thus be taken to eliminate such factors that may compromise hatching success. PMID- 18507218 TI - Comparison of quality of induction of anaesthesia between intramuscularly administered ketamine, intravenously administered ketamine and intravenously administered propofol in xylazine premedicated cats. AB - The quality of induction of general anesthesia produced by ketamine and propofol, 2 of the most commonly used anaesthetic agents in cats, was assessed. Eighteen cats admitted for elective procedures were randomly assigned to 3 groups and then premedicated with xylazine 0.75 mg/kg intramuscularly before anaesthesia was induced with ketamine 15 mg/kg intramuscularly (KetIM group), ketamine 10 mg/kg intravenously (KetIV group) or propofol 4 mg/kg intravenously (PropIV group). Quality of induction of general anaesthesia was determined by scoring ease of intubation, degree of struggling, and vocalisation during the induction period. The quality of induction of anaesthesia of intramuscularly administered ketamine was inferior to that of intravenously administered ketamine, while intravenously administered propofol showed little difference in quality of induction from ketamine administered by both the intramuscular and intravenous routes. There were no significant differences between groups in the ease of intubation scores, while vocalisation and struggling were more common in cats that received ketamine intramuscularly than in those that received intravenously administered ketamine or propofol for induction of anaesthesia. Laryngospasms occurred in 2 cats that received propofol. The heart rates and respiratory rates decreased after xylazine premedication and either remained the same or decreased further after induction for all 3 groups, but remained within normal acceptable limits. This study indicates that the 3 regimens are associated with acceptable induction characteristics, but administration of ketamine intravenously is superior to its administration intramuscularly and laryngeal desensitisation is recommended to avoid laryngospasms. PMID- 18507219 TI - A questionnaire survey of perceptions and preventive measures related to animal health amongst cattle owners of rural communities in KwaZulu-Natal, South Africa. AB - A questionnaire survey of 315 cattle owners from the rural districts of KwaZulu Natal was carried out. The aim of the survey was to improve our understanding of local farmers' perceptions and practices of animal disease prevention and control and to establish the extent of their relationship with veterinary services. The survey showed that many owners practice preventive measures such as deworming, tick control and vaccination. Traditional medicines were in use by over half the respondents (58.9 %). Diseases are regarded as an important management problem (56.1 %); ticks, worms and diarrhoea dominated the mentioned health problems in cattle. Veterinary services still play an important role and are a frequent source of advice to owners. The findings of the survey and their context are discussed. PMID- 18507220 TI - The effect of cholecystokinin peptides on ovine duodeno-jejunal slow waves with and without pretreatment with proglumide. AB - Cholecystokinin exerts a composite influence on gastrointestinal motility but little is known about its effect on small-intestinal slow waves. Thus, six rams were implanted with four bipolar serosal electrodes onto the duodeno-jejunal wall. In the course of chronic experiments the myoelectric activity was continuously recorded in the non-fasted animals. After recording of the full normal migrating myoelectric complex (MMC), 0.15 M NaCl or CCK peptides were injected intravenously during various phases of the next MMC cycle. Five ml of saline was injected over 30 s during phases 1, 2a, or 2b of the MMC. Cerulein was administered at doses of 1 (over 30 s), 10 (over 30 or 60 s), or 100 ng/kg (over 30, 60, 120 or 300 s) and cholecystokinin octapeptide (CCK-OP) at doses 20 times higher. CCK peptides were applied during early or late phase 1 of the MMC and during phases 2a and 2b of the MMC. In the course of additional experiments, saline and hormone administration was directly preceded by infusion of proglumide, an unspecific CCK receptor antagonist, at a dose of 10 mg/kg. The myoelectric recordings were continued until the arrival of a subsequent regular phase 3 of the MMC. In the duodenal bulb, slow waves were occasionally observed. In the duodenum the slow-wave frequency oscillated between 20 and 24 cpm and in the jejunum between 19 and 22 cpm before or after CCK peptides and proglumide. In the duodenum the slow-wave amplitude increased significantly after all doses of cerulein injected during phase 2b of the MMC. After administration of CCK-OP changes in duodenal slow-wave amplitude were not significant but exhibited a tendency similar to those after cerulein. In the jejunum, injection of cerulein and CCK-OP during phase 2 of the MMC increased the slow-wave amplitude significantly and the duration of these changes was longer than in the duodenum. After infusion of proglumide, administration of cerulein at the low dose over 30 s and at the high dose over 300 s in the course of late phase 1 and phases 2a and 2b of the MMC, significantly increased the duodenal slow-wave amplitude. Cerulein injection during phase 2b of the MMC at the high dose over 30 and 60 s, preceded by proglumide infusion, significantly inhibited the duodenal slow-wave amplitude. In the jejunum these changes were even more pronounced and their duration was much longer. It is concluded that CCK peptides affect slow-wave amplitude in the duodeno-jejunum in non-fasted sheep. This effect is stronger in the jejunum and is altered but not abolished by pretreatment with proglumide. Cerulein evokes more pronounced alterations in the slow-wave amplitude than CCK-OP in conscious sheep. PMID- 18507221 TI - The response of the pituitary-adrenal and pituitary-thyroidal axes to the plasma glucose perturbations in Babesia canis rossi babesiosis. AB - This prospective, cross-sectional, interventional study was designed to determine the association between the hormones of the pituitary-adrenal and pituitary thyroid axes and other clinical parameters with the blood glucose perturbations in dogs with naturally occurring Babesia canis rossi babesiosis. Thirty-six dogs with canine babesiosis were studied. Blood samples were obtained from the jugular vein in each dog prior to treatment at admission to hospital and serum endogenous adrenocorticotrophic hormone (ACTH), pre-ACTH cortisol, thyroxine, free thyroxine and TSH concentrations were measured. Immediately thereafter each dog was injected intravenously with 5 microg/kg of ACTH (tetracosactrin). A 2nd blood sample was taken 1 hour later for serum post-ACTH cortisol measurement. Three patient groups were recruited: hypoglycaemic dogs (glucose < 3.3 mmol/l, n = 12); normoglycaemic dogs (glucose 3.3-5.5 mmol/l, n = 12); hyperglycaemic dogs (glucose > 5.5 mmol/l, n = 12). Basal and post-ACTH serum cortisol concentrations were significantly higher in hypoglycaemic dogs, whereas body temperature, serum thyroxine and free thyroxine were significantly lower in hypoglycaemic dogs. Haematocrit was significantly lower in both hypo-and hyperglycaemic dogs compared with normoglycaemic dogs. Low blood glucose concentrations were significantly associated with high basal and post-ACTH cortisol concentrations and with low serum thyroxine and free thyroxine concentrations in dogs suffering from B. canis rossi babesiosis. PMID- 18507222 TI - Multiple cutaneous inverted papillomas in a dog. AB - Cutaneous inverted papillomas are described in an 8-month-old mixed breed domestic dog from Windhoek, Namibia. Multiple firm, rapidly growing, doughnut shaped masses formed on the ventral abdomen, which histologically consisted of a cup-shaped rim of marked epithelial hyperplasia, giant keratohyaline granules and prominent koilocytes and marked hyperkeratosis filling the centre of the mass. Current literature on canine papillomas is briefly reviewed. PMID- 18507223 TI - Periodontics: 10. Maintenance in periodontal therapy. AB - Maintenance periodontal therapy is essential for the long-term stabilization of periodontal disease. An individualized programme of maintenance requirements, based on patient risk assessment for disease progression, is important, as is the need for teamwork in long-term patient management. CLINICAL RELEVANCE: Maintenance or supportive therapy is an essential requirement in the management of periodontal therapy. Patients should be informed at their first visit that part of the management of their disease is regular maintenance visits. PMID- 18507224 TI - Facial aesthetics: 2. Clinical assessment. AB - The clinical ability to alter dentofacial form requires an understanding of facial aesthetics. This is vital for any clinician involved in treatment that will alter a patient's dentofacial appearance, whether through orthodontics, facial growth modification, corrective jaw surgery or aesthetic dentistry. Part 1 of this article covered the historical and theoretical aspects of facial aesthetics and their importance in contemporary dentofacial treatment. Part 2 covers important aspects of the interview and clinical assessment of patients requiring alterations in their dentofacial appearance, including guidelines used in the assessment of facial proportions and symmetry. CLINICAL RELEVANCE: These articles cover the theoretical and clinical aspects of facial aesthetics required by clinicians involved in the treatment of dentofacial deformity. PMID- 18507225 TI - Influence of fractured instruments on the success rate of endodontic treatment. AB - The fracture of an instrument is a recognized complication in endodontics. The immediate response to a fractured instrument is frequently to regard the treatment as a failure. Several factors must, however, be taken into account to evaluate the prognosis of the tooth in this situation. The objective of the endodontic treatment with or without a fractured instrument remains the same, namely to disinfect the root canal system and prevent its recontamination. The time at which file fracture occurred during treatment and the degree of canal infection should be considered when determining the potential effect of instrument fracture on treatment outcome. Patients must be informed about an instrument fracturing in their tooth for ethical and legal reasons. The aim of this paper is to attempt to place fractured instruments in context, not to provide an in depth description of fractured instrument management techniques. CLINICAL RELEVANCE: To understand the influence of fractured instruments on prognosis in endodontics. PMID- 18507226 TI - The successful management of complaints--turning threats into opportunities. AB - This article identifies the drivers that influence a patient's decision to complain about his/her care and the multifactorial reasons for this. It explores the effects complaints have on dentists and their teams and how these reactions should not be underestimated, even when the complaint can be quite minor. The article goes on to give some generic advice about complaints handling with a brief overview of the NHS procedures in the new contract and finishes with some useful strategies to prevent complaints occurring in the first place. CLINICAL RELEVANCE: Dealing with complaints is an important and ever increasing part of dental practice. If handled well, the patient will continue his/her relationship with the practice and lessons will be learnt. These lessons can be both clinical and non-clinical but, in many cases, complaints can be prevented by effective communication skills. PMID- 18507227 TI - Selecting appropriate recall intervals for patients in general dental practice- an audit project to categorize patients according to risk. AB - In 2004, the National Institute for Health and Clinical Excellence (NICE) was asked to prepare guidance for the NHS in England and Wales on the clinical need and cost-effectiveness of a dental recall examination for all patients at an interval based upon their risk from oral disease. (Dental Recall: recall interval between routine dental examinations). Unlike other NICE guidelines, this potentially applies to the whole population. The Department of Health wishes to reduce the number of'routine' six-monthly check-ups. Best practice now recommends that all patients undertake an oral health assessment prior to categorizing them into a recall programme. The article describes how an audit project was undertaken to establish compliance within General Dental Practice to the guidelines. CLINICAL RELEVANCE: Dentists and patients now have the opportunity to discuss appropriate recall intervals together and prescribe reviews at time periods appropriate to each patient's disease process and risk. PMID- 18507228 TI - Management of the petrified dental patient. AB - Fear and anxiety are common responses to anticipated or actual dental treatment. This may range from slight feelings of unease during routine procedures, such as an injection or cavity preparation, to feelings of extreme anxiety long before treatment is happening. There is general agreement that anxiety constitutes a major problem for the dentist, and a barrier to optimal care for the patient, as one of the most significant consequences is avoidance of dental treatment. Common triggers include local anaesthetic injection and the dental drill. This paper uses the term anxiety to encompass both fear and anxiety and reviews current strategies for managing anxious people in the dental setting, with a focus on non invasive operative techniques. CLINICAL RELEVANCE: Being able to recognize fear and anxiety in dental patients is an important part of overall patient management. Knowledge of various behaviour management techniques, and the use of less-invasive operative techniques, enable the clinician to treat a large proportion of anxious people and help them to cope with dental treatment in the primary care setting. Those who remain anxious and unable to cope in the dental setting may require pharmacological techniques, such as conscious sedation, to enable them to receive dental treatment. These techniques are not always readily available, require appropriate training, are more time consuming and costly. In extreme cases, general anaesthesia may be a last resort option. PMID- 18507229 TI - Making it stick. PMID- 18507230 TI - EACD GDP guidelines. AB - The recommendations for examination, diagnosis and management of patients with temporomandibular disorders and orofacial pain by the general dental practitioner published by the EACD do not provide a step by step guidance for the GDP; a training programme would be needed for that. They provide a general framework under which these conditions should be diagnosed and treated. The evidence base for the advice given is presented. In particular, they advise against some unproven and invasive treatments. PMID- 18507231 TI - Physical signs for the general dental practitioner. Case 53. Medical conditions in dental practice. Angina pectoris. PMID- 18507232 TI - Aspects of human disease. 21. Anxiety, stress and depression. PMID- 18507233 TI - Quality healthcare for America's children. PMID- 18507234 TI - The quality of pediatric care--how will we cross the chasm? PMID- 18507235 TI - A home for medically complex children: the role of hospital programs. AB - The medical home is a conceptual model of continuous and comprehensive care provision that is associated with improved outcomes for children with special healthcare needs. Most applications of the medical home have focused on improving primary care services. Despite concerted efforts to apply the medical home concept to the care of children with special healthcare needs, many barriers to its implementation still exist, in particular, for the subset of children with special healthcare needs who are medically complex. Applying the medical home concept to hospital-based care coordination may benefit both children with complex conditions and their families, as well as the community-based providers. PMID- 18507236 TI - Evaluation of standardized teaching plans for hospitalized pediatric patients: a performance improvement project. AB - Discharge teaching in a pediatric hospital setting is difficult because the situation involves multiple learners, time constraints, and differing skill levels of nurse teachers. Shortened length of stay forces nurses to complete patient education efficiently. Unstructured education can lead to failed learning, as evidenced by readmissions and postdischarge feedback. A performance improvement project was conducted to evaluate the effectiveness of standardized teaching plans for diabetes mellitus and infant cardiopulmonary resuscitation. Preliminary data indicated a passing score of at least 90% on posttests by all learners, suggesting that standardized teaching plans may help nurses complete prescribed discharge teaching. PMID- 18507237 TI - Condition HELP: a pediatric rapid response team triggered by patients and parents. AB - Condition HELP is a unique pediatric patient safety program that gives families an immediate voice in their child's medical care. This patient safety endeavor allows parents to call a telephone number at any time-24 hours a day, 7 days a week-if they feel their hospitalized child's immediate health is endangered. The Condition HELP call receives the same immediate attention as a life-threatening emergency, including deployment of a specialized rapid response team to the child's bedside. This article presents a 2-year analysis of pediatric patient safety outcomes following the implementation of the program. PMID- 18507238 TI - Interview with a quality leader: Barbara Spreadbury on children's health. Interview by Christy L. Beaudin. PMID- 18507239 TI - Journey toward meaningful pediatric quality metric reporting: the Texas experience. AB - Working under a mandate for public reporting, children's hospitals in Texas joined in a partnership with the state with the intent of working toward providing meaningful assessment of the quality of pediatric inpatient care. This article summarizes a journey of nearly 2 years undertaken to review currently available quality measures and arrive at interagency consensus for the reporting of pediatric quality and clinical outcomes in Texas. Public reporting has been approached with great divergence across the states. The Texas project underlines the need for all interested parties to collaborate for best results. PMID- 18507240 TI - A novel error-reporting tool in pediatric intensive care. AB - Patient safety is a critical component of the U.S. healthcare system: thousands of people, including children, die or are injured yearly as a result of medical error. We designed and implemented a novel error-reporting tool for the pediatric intensive care unit. More errors were reported with the use of this paper-based tool than with the existing computerized error-reporting system. We also developed a scoring system to assess potential harm to the patient. The tool provided information about frequent and high-risk errors that guided successful improvements in patient care and safety and the achievement of measurable success. PMID- 18507241 TI - National Quality Forum issue brief: strengthening pediatric quality measurement and reporting. AB - National efforts to spur quality improvement through public reporting and pay for performance have to date focused largely on the adult population, but now there is growing interest and momentum to include pediatric measures in the effort to promote accountability and improvement in the care that is delivered to America's children. Four leading national pediatric organizations recently came together to form the Alliance for Pediatric Quality (the Alliance), an organization that works to identify and encourage the adoption of pediatric quality measures suitable for quality improvement and public reporting. The National Quality Forum (NQF) already has endorsed more than a dozen quality measures specific to pediatric care, and another two dozen pediatric measures currently are under review. Together, these efforts reflect increased resolve to bring pediatric care into the fold of pay-for-performance and public reporting initiatives, and they offer much promise for driving rapid improvements in the quality of healthcare for children. PMID- 18507242 TI - Institute for Healthcare Improvement's 19th aNnual National Forum on Quality Improvement in Health Care. PMID- 18507243 TI - The Joint Commission's latest white paper urges development of a national performance measurement data strategy. PMID- 18507244 TI - APPROVED: approach for establishing target measure ranges for performance measure reporting. PMID- 18507245 TI - Sentinel event alert: preventing pediatric medication errors. PMID- 18507246 TI - REMINDER: proficiency testing practices for laboratories. Laboratories must take steps to avoid even the appearance of referral. PMID- 18507247 TI - Journeys: stories of nurses' careers. PMID- 18507248 TI - The 40-year-old health care virgin: I did it my--highly unorthodox--way. PMID- 18507249 TI - My journey in nursing. PMID- 18507250 TI - My journey in nursing. PMID- 18507251 TI - Many paths in nursing: one nurse's story. PMID- 18507253 TI - Looking ahead to a new generation of Creative Nursing. PMID- 18507252 TI - My journey. PMID- 18507254 TI - Tradition, legacy, attitude and obstinacy: right or wrong, survival involves transformation. PMID- 18507255 TI - The future of TMA: where does Tennessee's largest physician organization go from here? PMID- 18507256 TI - Too much to do and so little time: chest pain with syncope. PMID- 18507257 TI - Supply side change... More to it than meets the eye! PMID- 18507258 TI - Improving physician efficiency and patient convenience via online lab results. PMID- 18507259 TI - Relationship between health literacy and health-related quality of life among Tennesseans. AB - The purpose of this study was to examine the relationship between health literacy and health-related quality of life (HRQOL) in a sample of English-speaking Tennesseans attending a university-based family medicine residency clinic. We measured the health literacy skills of patients (n = 249) using the Rapid Estimate of Adult Literacy in Medicine (REALM), dividing patients' scores into limited (< or = 8th-grade level) and adequate (> or = 9th-grade) literacy skill levels. We measured HRQOL using four items developed and validated by the Centers for Disease Control and Prevention (CDC). Scores on the REALM indicated that 28.5 percent of patients had limited and 71.5 percent had adequate health literacy skills. There was a significant correlation between patients' health literacy skills and self-reported general health (rho = 0.249, P < 0.01). Patients with limited health literacy skills had a higher median number of physically unhealthy days (P = 0.05) and activity limitation days (P = 0.05) compared with patients with adequate health literacy skills. Based on this study, patients' health literacy skills are associated with several components of HRQOL. PMID- 18507260 TI - Expert testimony in medical malpractice litigation and use of "magic words". PMID- 18507261 TI - History of Tennessee medicine, Pt. XII--first survey of Tennessee physicians. PMID- 18507262 TI - Racial disparities in trends for cardiovascular disease and procedures among hospitalized diabetic patients. AB - METHODS: To determine if racial differences exist for trends in diabetes-related cardiovascular disease (CVD) hospitalization rates, we analyzed data from an inpatient hospital discharge database maintained by the South Carolina Office of Research and Statistics. All hospitalizations involving a diagnosis of diabetes were collected from 1996 through 2003. International Classification of Diseases codes were used to determine diagnosis for diabetes, acute myocardial infraction (AMI), stroke, and other CVD outcomes. Multiple linear regression was performed to model the age-standardized rates during the study period. An interaction parameter for race and discharge year was used in the models to determine if the trend slopes varied between African Americans and Caucasians. RESULTS: The diabetes-related hospitalization rates for AMI and stroke declined for both race groups. Although the stroke rates for African Americans were consistently higher than those for Caucasians, the African American trend declined more sharply (P=.027). AMI rates showed sharper declines among Caucasians (P<.001). Rates of CVD procedures (percutaneous transluminal coronary angioplasty and coronary artery bypass graft) were two to three times greater among Caucasians. Cardiomyopathy rates were significantly greater among African Americans and showed a larger increasing trend (P<.001), and findings for congestive heart failure trends were similar (P<.001). CONCLUSIONS: Diabetes-related CVD rates and trends vary considerably by race. Rates of AMI and stroke declined in African Americans and Caucasians from 1996 through 2003, while other CVD rates increased. Further research is needed to understand the underlying components of these disparities. PMID- 18507263 TI - Identification of diabetic complications among minority populations. AB - OBJECTIVE: In consideration of the increasing prevalence of diabetes, multiple factors related to levels of long-term glycemic control, and complex causes of racial disparities across a variety of chronic conditions, patterns of admissions and complications related to diabetes by ethnicity were explored to develop a more clear understanding of underlying causes of disparities. METHOD: Using the 2003 National Inpatient Sample, we analyzed the correlation between the primary diagnosis and the likelihood that the condition represented poorly controlled diabetes or a diabetes-related complication. RESULTS: Minorities were more likely to be admitted through the emergency department and for a condition directly related to diabetes progression. Further, minorities were more likely to be admitted for acute hyperglycemia and acute hypoglycemia. CONCLUSION: Interventions that address root causes of disparities related to diabetes and other conditions, such as care-seeking behaviors and ease of access to primary care providers, are keys to eliminating ethnic disparities. PMID- 18507264 TI - Grandparenthood status and health outcomes in midlife African American women with type 2 diabetes. AB - OBJECTIVE: To describe grandparenthood status and determine the relationship between grandparenthood status, metabolic control, and psychosocial factors in African American women with type 2 diabetes. METHODS: A secondary, descriptive, exploratory analysis was conducted to examine the relationship between grandparenting status, physiologic indices related to metabolic control (body mass index, hemoglobin A1c, systolic blood pressure, low-density lipoprotein cholesterol) and psychosocial factors (emotional distress and quality of life) in a sample of 109 urban midlife African American women with type 2 diabetes. RESULTS: Grandmothers made up 60% of the sample and were stratified into three groups on the basis of caretaking status. Grandmothers who cared for but did not live with grandchildren had the highest body mass index, hemoglobin A1c, low density lipoprotein cholesterol, and systolic blood pressure. Caregiving grandmothers also had higher diabetes-related emotional distress scores but had better quality of life scores than did the non-caretaking grandmothers. Non grandmothers had the lowest quality of life scores. CONCLUSIONS: These findings suggest relationships between grandmother caretaking status and metabolic and psychosocial factors. Research is needed to further examine these relationships and implications on practice and policy decisions. PMID- 18507265 TI - Hypomagnesemia in Ethiopians with diabetes mellitus. AB - BACKGROUND: Magnesium, the second most abundant intracellular cation, plays a key role in cellular metabolism. Even though hypomagnesemia has been demonstrated in patients with both type 1 and type 2 diabetes elsewhere, limited information exists from African patients with diabetes mellitus. METHODS: This was a cross sectional study to assess the prevalence of hypomagnesemia in Ethiopian patients with type 1 and type 2 diabetes. A total of 159 subjects were included in the study (44 patients had type 1 diabetes, 69 patients had type 2, and 46 were nondiabetic controls). RESULTS: The mean age (+/-standard error of the mean) of the subjects with type 1 diabetes, type 2 diabetes, and controls was 30.6+/-1.6, 51.3+/-1.3, and 29.0+/-1.7 years, respectively (P<.001). Patients with type 2 diabetes were significantly older than those with type 1 diabetes and controls. Basal C-peptide level and body mass index were also significantly higher in patients with type 2 diabetes (P<.001). The mean magnesium level was significantly lower in patients with diabetes than in controls (.84+/-.12 mmol/L vs 1.02+/-.17 mmol/L, P<.001). Hypomagnesemia was seen in 65% of the patients with diabetes. CONCLUSION: The study showed that patients with diabetes mellitus have lower levels of magnesium and are therefore at increased risk of complications related to magnesium. In light of these potential complications, we recommend periodic determination of magnesium levels and appropriate magnesium replacements. PMID- 18507266 TI - Resistance training differentially affects weight loss and glucose metabolism of White and African American patients with type 2 diabetes mellitus. AB - OBJECTIVE: We aimed to identify whether racial differences in body composition and glucose metabolism occur in response to exercise and determine whether aerobic and resistance exercise modalities bring about differential changes in these parameters in African Americans and White persons with type 2 diabetes. RESEARCH DESIGN AND METHODS: Participants included 36 African American and 23 White men and women with type 2 diabetes who were randomly assigned to eight weeks of either resistance or aerobic exercise. Before and after this intervention, each participant underwent a series of measurements that assessed anthropometrics and glucose metabolism. RESULTS: African Americans responded more favorably to resistance training than did Whites. This difference was manifested by a significant improvement in BMI (-2.57%+/-.90% vs 2.57%+/-1.09%, P<.01) and insulin resistance (-19.15%+/-9.00% vs 13.12%+/-11.86%, P<.05) in African Americans compared to Whites. When comparing exercise modalities within the races, African Americans demonstrated a preferential response to resistance training. CONCLUSIONS: Eight weeks of resistance training by African Americans may have a more positive effect on weight loss and glucose metabolism than aerobic exercise training. Furthermore, the changes observed appear to be unique to African Americans, as no changes were observed in Whites after an equal amount of resistance training. When an exercise program is designed for a person with type 2 diabetes, race should be taken into consideration, and resistance exercise for African Americans may lead to increased weight loss and improved insulin sensitivity than does aerobic exercise. PMID- 18507267 TI - Multiple health behaviors among overweight, class I obese, and class II obese persons. AB - OBJECTIVES: Recent studies suggest an association between excess weightand increased risk of some cancers. Health disparities are evident for both obesity and cancer, each of which disproportionately affects African American adults. We examine the relationship between weight and selected health behaviors related to colorectal cancer (CRC) prevention (fruit and vegetable consumption, recreational physical activity, and CRC screening). We also examine behavioral psychosocial correlates including knowledge, perceived benefits and barriers, self-efficacy, and social support for these behaviors. METHODS: The WATCH (Wellness for African Americans through Churches) Project was a CRC prevention study implemented in African American churches in rural North Carolina. We analyzed the baseline data of 813 church members who provided information on their height and weight through a telephone-based survey. RESULTS: Most (78%) respondents were classified as overweight or obese. Self-rated health and level of physical activity were lower at higher weight levels, but little difference in fruit and vegetable consumption was observed among participants. Weight was negatively associated with past-year CRC testing among women but not among men. Levels of knowledge and self-efficacy were similar across weight groups, but some perceived barriers were significantly higher among obese participants. CONCLUSIONS: Obesity was associated with some health behaviors and psychosocial correlates associated with increased cancer risk. Cancer prevention programs in African American populations where overweight is prevalent may wish to specifically address these issues. PMID- 18507268 TI - Anthropometric correlates of metabolic syndrome components in a diverse sample of overweight/obese women. AB - OBJECTIVE: The purpose of this study was to determine the relationship between body mass index (BMI), waist circumference, and waist-to-hip ratio (WHR) with cardiometabolic variables that reflect the metabolic syndrome in overweight/obese premenopausal White, African American, and Hispanic women. METHODS: Two hundred and thirty four young overweight/obese women enrolled in a weight loss program were recruited for this study. Analysis of variance was used to compare means among groups, and multiple regression analyses were used to determine the relationship between anthropometric measurements and cardiometabolic variables, after controlling for age. RESULTS: In both White and African American women, BMI was significantly related to systolic blood pressure and diastolic blood pressure, while in Hispanic women, BMI failed to predict any cardiometabolic variables. Using waist circumference afforded the additional prediction of high density lipoprotein cholesterol (P=.017) and triglycerides in White women and serum glucose in African American women. In Hispanic women, waist circumference significantly predicted serum glucose. WHR was the strongest predictor of metabolic syndrome components in White women; however, it failed to predict any cardiometabolic variables in Hispanic women. CONCLUSIONS: Both waist circumference and WHR were better correlates of metabolic syndrome components than was BMI. While WHR appeared optimal for predicting components of the metabolic syndrome in White women, our findings showed that waist circumference was the most global anthropometric indicator of metabolic syndrome components in a diverse racial and ethnic sample of overweight/obese women. PMID- 18507269 TI - Sustainability of a multiple risk factor intervention on cardiovascular disease in high-risk African American families. AB - OBJECTIVES: To determine the long-term effect of a community-based risk reduction intervention at five years after completion of a one-year randomized clinical trial and to determine the sustainability of the beneficial effects seen one year after the intervention. METHODS: 30- to 59- year-old African American siblings of probands with premature coronary heart disease (CHD) were randomized for care of multiple CHD risk factors to either one year of community-based care (CBC) provided by a nurse practitioner/community health worker team or enhanced usual care (EUC). At five years, 307 (84.6%) of the siblings returned for reevaluation. MAIN OUTCOME MEASURES: Changes in and achievement of goal levels of low-density lipoprotein cholesterol (LDL-C), systolic and diastolic blood pressure (SBP and DBP, respectively), and smoking cessation at five years. RESULTS: No significant differences were seen between groups in mean LDL-C, SBP, and DBP or in the overall percentages achieving goal LDL-C, blood pressure, or smoking status. Changes after completion of the intervention suggest that the CBC group lost the beneficial effects for mean LDL-C and for percentage at goal LDL-C, while the EUC group continued to improve. CBC was associated with greater sustainability and less refractoriness of one-year results for LDL-C and blood pressure goals. CONCLUSIONS: Although no group differences were found in mean risk factor levels at five years, data indicate that CBC is both feasible and associated with earlier sustainability of positive risk factor changes compared with EUC. PMID- 18507270 TI - Racial differences of lipoprotein subclass distributions in postmenopausal women. AB - BACKGROUND: We assessed racial differences in lipoprotein particle size, a marker of atherosclerosis risk, among women with coronary disease. METHODS: We studied 378 women (33% non-White, predominantly African American) at the baseline visit of the Women's Angiographic Vitamin and Estrogen Trial (WAVE), a multicenter trial of hormone replacement and antioxidant vitamin therapy in postmenopausal women with established coronary artery disease. Average particle sizes for high density lipoprotein (HDL), low-density lipoprotein (LDL), and very low-density lipoprotein were measured by nuclear magnetic resonance in these women, and angiography was performed at baseline and followup. RESULTS: Adjusted for age, race, diabetes, smoking, blood pressure, and use of lipid-lowering and antihypertensive medications, non-White women had larger LDL particle size (difference .2 nm, 95% CI .1-.3 nm) and HDL particle size (difference.2 nm, 95% CI .1-.2 nm). Neither angiographic disease progression nor survival without myocardial infarction (median follow-up time of 2.8 years) was associated with lipoprotein particle size or race. CONCLUSIONS: Non-White women have a less atherogenic profile of lipoprotein particle sizes than do White women. However, this difference did not affect event-free survival or angiographic progression of coronary atherosclerosis. PMID- 18507271 TI - Blood pressure and memory in older African Americans. AB - The rates of high blood pressure among African Americans, as a group, are the highest in the world. The implications for higher average blood pressure include complications for many major chronic conditions, such as cardiovascular disease and diabetes. Less well studied is the effect of blood pressure on the cognitive functioning of African Americans. The purpose of this study was to examine the effect of blood pressure on memory measures in a sample of adult African Americans. Analyses were conducted on a sample of 361 African American adults (mean age 61.50 years, standard deviation 9.39 years). We found significant correlations between systolic blood pressure and most cognitive measures but only for one of the measures and diastolic blood pressure. Regressions revealed significant effects for systolic blood pressure on Digit Symbol, Telephone Interview of Cognitive Status, and Immediate Recall on the Wechsler Logical Memory test. These findings suggest that blood pressure is a source of individual variability in cognitive aging among African Americans. PMID- 18507272 TI - The influence of cardiac and vascular responses on baseline cardiovascular parameters in Black African children. AB - BACKGROUND: The prevalence of hypertension among Black adults in South Africa is high. Because cardiovascular reactivity can be used to predict hypertension in later life, we attempted to determine whether cardiac and vascular responses to a task in Black South African children influence their cardiovascular parameters. DESIGN: This study was embedded in the Transition and Health during Urbanization in South Africa in Children study, which studied the health status of children. During the study, cardiovascular reactivity was determined with a hand dynamometer in 670 Black African children. Systolic blood pressure, diastolic blood pressure, mean arterial pressure, heart rate, stroke volume, cardiac output, total peripheral resistance, and Windkessel compliance of the arterial system were obtained by means of the Finapres (finger arterial pressure) apparatus and the Fast Modelflow software program. Anthropometric measurements were performed according to standard methods. Cardiac and vascular responders were classified with a regression-based approach. RESULTS: Cardiac responders had higher stroke volume; a trend to higher cardiac output; lower diastolic blood pressure; and a tendency toward lower systolic blood pressure, heart rate, and total peripheral resistance. Vascular responders showed no significant changes in cardiovascular parameters when responders and nonresponders were compared. CONCLUSIONS: Cardiac responders had higher stroke volume and a trend toward higher cardiac output, which may be an early indication of hypertension. PMID- 18507273 TI - Racial and ethnic disparities associated with knowledge of symptoms of heart attack and use of 911: National Health Interview Survey, 2001. AB - OBJECTIVE: Heart attacks are more prevalent among Hispanics and Blacks than among Whites. Bystanders must be able to recognize heart attack symptoms and activate the emergency response system in order to receive time-dependent therapies that increase survival. This study estimated racial/ethnic disparities in awareness of heart attack symptoms in a sample of the US population. METHODS: We evaluated data from 33,059 adult participants in the 2001 National Health Interview Survey. Respondents indicated their awareness of five heart attack symptoms and the need to call 911 in the presence of such symptoms. RESULTS: Hispanics and Blacks were less likely to recognize each heart attack symptom than were Whites (P<.05). Hispanics (25.6%), people aged 18-24 years (33.6%), men (39.1%), and those with less than a high school education (31.3%) were less likely to recognize all five heart attack symptoms and report that they would call 911 than were Whites (45.8%), Blacks (36.1%), respondents aged 45-64 years (47.7%) and >65 years (43.9%), and those with a high school education (41.0%) or more (45.6%). In multivariate logistic regression analyses, Blacks (OR .73, 95% CI .66-.80) and Hispanics (OR .49, 95% CI .45-.54) were less likely than were Whites to recognize all five heart attack symptoms and the need to call 911 if someone had these symptoms. CONCLUSIONS: One Healthy People 2010 goal is to eliminate health disparities. Racial/ethnic disparities exist in knowledge of heart attack symptoms and the need to call 911. Special educational efforts should focus on Black and Hispanic populations and highlight the importance of symptoms and time dependent therapies. PMID- 18507274 TI - Racial/ethnic differences in stroke awareness among veterans. AB - OBJECTIVE: To examine racial/ethnic differences in stroke recognition and knowledge of appropriate first action if someone was having a stroke. METHODS: We examined data from 36,150 veterans from the 2003 Behavioral Risk Factor Surveillance System (BRFSS). Respondents indicated recognition of five stroke warning signs/symptoms and first action they would take if someone were having a stroke. Multiple logistic regression was used to assess the independent effect of race/ethnicity on stroke recognition and appropriate first action, controlling for relevant covariates. RESULTS: Most respondents recognized at least one warning sign: 96% recognized sudden confusion or trouble speaking; 97% recognized sudden facial, arm, or leg weakness; 88% recognized sudden vision loss; 94% recognized sudden trouble walking; and 80% recognized sudden headache; 86% recognized calling 911 as the appropriate first action. However, only 17% recognized all five warning signs/symptoms, and only 15% recognized all five warning signs/symptoms and would call 911 as the first action. In multivariate models with Whites as reference, Hispanics (OR .34, 95% CI .22-.51) and Others (OR .68, 95% CI .50-.92) were less likely to recognize all five stroke warning signs/symptoms. Hispanics (OR .37, 95% CI .24-.58) and Others (OR .68, 95% CI .48 .96) were less likely to recognize all five warning signs/symptoms and call 911 as the first action. CONCLUSIONS: Most veterans recognize individual stroke warning signs, but very few recognize all five and would take appropriate action to call 911 in the event of a stroke. Low rates of stroke recognition and taking appropriate action are more pronounced in racial/ethnic minority veterans. PMID- 18507275 TI - Racial differences in medication compliance and healthcare utilization among hypertensive Medicaid recipients: fixed-dose vs free-combination treatment. AB - OBJECTIVE: To assess compliance with antihypertensive therapy and healthcare utilization among African American and White Medicaid recipients who are receiving fixed-dose combination amlodipine besylate/benazepril HCl or a dihydropyridine calcium channel blocker plus an angiotensin-converting enzyme inhibitor prescribed as separate agents (free-combination). DESIGN: Longitudinal, retrospective, cohort analysis of South Carolina Medicaid claims for the years 1997 through 2002. Followup was 12 months from the index date, defined as the first prescription dispensing date for a study drug. SETTING AND PARTICIPANTS: South Carolina Medicaid beneficiaries receiving fixed-dose (n=3363) and free combination (n=713) therapy, including 3016 African Americans and 1060 White patients. MAIN OUTCOME MEASURES: Compliance was defined as the total days' supply of drug (excluding last prescription fill) divided by the length of followup; healthcare utilization included cost and number of claims associated with ambulatory services, hospital care, and prescription drugs. RESULTS: The cohort (N=4076) was 74.0% African American; mean age was 62.2 years. Compliance was significantly greater in patients who received fixed-dose therapy than in those who received free-combination therapy (58.6% vs 48.1%; P<.05). The average total cost of care was lower for the fixed-dose group ($4605) than for the free combination group ($8531). African Americans and Whites were equally likely to receive the fixed-dose combination. However, compliance was lower among African American patients than among White patients (55% vs 61% respectively; P<.05). Costs and claims for ambulatory and hospital services were higher for African American patients, whereas drug costs and claims were higher for White patients. CONCLUSION: Fixed-dose amlodipine besylate/benazepril HCl was associated with higher compliance rates than was free-combination therapy, independent of race. Lower compliance rates among African American patients may have contributed to the higher healthcare resource use and costs observed. Efforts to enhance medication compliance tailored to African Americans may improve outcomes and reduce costs in this high-risk population. PMID- 18507276 TI - Barriers to cancer clinical trial participation among Native elders. AB - OBJECTIVES: American Indians/Alaska Natives are underrepresented in clinical trials. Therefore, they must participate in large-scale cancer clinical trials to ensure the generalizability of trial results and improve their access to high quality treatment. Our goal was to identify factors that influenced participation in cancer clinical trials among American Indians/Alaska Natives. METHODS: An anonymous survey that assessed willingness to participate in a hypothetical cancer clinical trial and how 37 factors influenced their willingness to participate was administered to 112 older American Indian/Alaska Native adults at an annual social event honoring elders. Responses ranged from one (definitely would not participate) to five (definitely would participate). Data were analyzed with ordinal logistic regression. RESULTS: Factors that most strongly increased willingness to participate were having a lead researcher of Native descent, having a study physician with experience treating American Indians/Alaska Natives, personal experience with the cancer being studied, family support for participation, and belief/hope that the study would result in new treatments. Factors that decreased willingness to participate most strongly were living far from the study site and a high risk that confidentiality could be breached. CONCLUSIONS: Our results identify conventional and culturally unique barriers to research participation among older American Indians/Alaska Natives. These data emphasize the need to establish partnerships with Native communities and include American Indian/Alaska Native and culturally competent professionals in research efforts. Of equal importance are disseminating information about clinical trials and recognizing the role of family in decisionmaking in this group. PMID- 18507277 TI - Parental occupation, Hispanic ethnicity, and risk of selected congenital malformations in offspring. AB - OBJECTIVES: Evidence suggests that parental occupation and Hispanic ethnicity may be risk factors for some birth defects. Because few studies have examined the effect of Hispanic ethnicity on occupational associations, we examined whether risk associated with certain occupations was heightened in Hispanics compared with non-Hispanic Whites. DESIGN: In this case-control study among Texas births occurring from 1996 through 2000, cases of neural tube defects, isolated oral clefts, and chromosomal anomalies were linked to their respective live birth certificates. A random sample of 4965 live births without documented congenital malformations served as the comparison group. Parental occupations were categorized into groups according to previously published associations. Logistic regression was used to obtain odds ratios (OR) and 95% confidence intervals (CI) for the selected congenital malformations in relation to parental occupations. RESULTS: Maternal occupations as cook or nurse were associated with oral clefts (OR 3.3, 95% CI .6-16.0) and neural tube defects (OR 3.1, 95% CI .5-13.1), respectively, among births to Hispanic mothers, but not with births to non Hispanic White mothers. Hispanic fathers who were electricians were more likely to have offspring with chromosomal anomalies, especially trisomy 18 (OR 7.4, 95% CI 1.6-25.5), associations not seen among offspring of non-Hispanic White fathers. Risk estimates also differed by Hispanic ethnicity between oral clefts and paternal occupations of electronic equipment operator, farmworker, janitor, police officer, and printer. CONCLUSIONS: In this study, we found differences for risk of several congenital malformations by Hispanic ethnicity in relation to parental occupation. We recommend further study of these risks in other Hispanic populations. PMID- 18507278 TI - Racial influences associated with asthma management among children in the United States. AB - OBJECTIVE: To examine the influence of race and having an asthma management plan on the impact of experiencing asthmatic episodes. METHODS: This study utilized the 2002 and 2003 National Health Interview Survey to conduct a retrospective study and secondary data analysis. Univariate, bivariate, and multivariate analysis was performed to examine physician asthma management plan recommendations among minority and non-minority children in the United States. RESULTS: Most of the study participants (59%) reported not having an asthma management plan. Children who experienced an asthma episode in the past 12 months were less likely to have an asthma management plan (OR .51, P<.0001). In the multivariate analysis, Whites were significantly more likely than were Blacks and Hispanics to have an asthma management plan (OR 1.66, P=.0031). CONCLUSIONS: Findings from this study indicate that Black and Hispanic children with asthma are less likely to have an asthma management plan, and children with an asthma management plan are less likely to have had asthma episodes in the past 12 months. Requiring all insurers to provide an asthma management plan to children with asthma may reduce these race-based inequities. PMID- 18507280 TI - Patterns of blood flow may predict high blood pressure in later life. PMID- 18507279 TI - Biological risk factors relevant to chronic disease in three ethnic groups in Taiwan: results from Li-Shin Outreaching Neighborhood Screening (LIONS A1). AB - OBJECTIVES: Few population-based studies have compared risk factors related to chronic diseases across multiethnic groups of Chinese people. Consequently, we report the prevalence of common chronic disorders that make up the metabolic syndrome and compare their distribution in three ethnic subgroups: the offspring of Hakka, Minnan, and mainlander women. METHODS: We included 6854 participants in the Li-Shin Outreaching Neighborhood Screening (LIONS) project: 3088 (45.1%) Hakkas, 2461 (35.9%) Minnans, and 1305 (19.0%) mainlanders. Information on demographic features and recognized lifestyle factors was collected by using questionnaires; data on biological markers of metabolic syndrome were collected from serum samples by using standard biochemical analyses. RESULTS: Miscegenation averaged 22%. Smoking, alcohol consumption, and betel chewing varied across the three subpopulations. After controlling for demographic features and these three risk factors, men with mainlander mothers had more body fat. Compared with offspring with Hakka mothers, attendees whose mothers were from Minnan had higher uric acid concentrations. CONCLUSIONS: Despite the rarity of racial miscegenation in the three ethnic groups, most biological markers of metabolic syndrome were identical across the groups. Disparities were found for hyperuricemia in attendees whose mothers were from Minnan and for obesity in men whose mothers were mainlanders. These findings can help design health policy for the early detection of chronic disease in different ethnic Chinese groups. PMID- 18507281 TI - Encouraging American Indians, Alaska Natives to participate in cancer clinical trials. PMID- 18507282 TI - Know the signs of stroke and steps to take to help a stroke victim. PMID- 18507283 TI - Asthma management plans for children can lead to a healthier life. PMID- 18507284 TI - Assessing the risk to U.K. children from carbendazim residues in apple products. AB - We present a detailed study into the effects that uncertainty and variability have on the levels of dietary exposure of young children to carbendazim in a range of apple products, including raw apples, apple juice and processed apple foods. It is estimated that exposure is greater than the provisional EU acute reference dose of 0.02 mg/kg-bw/day on 0.29% of person days, although there is large uncertainty in the true number. The contribution from apple juice consumption appears to be significant. The issues presented are common to many dietary risk analysis problems, and the methods could be applied to any food group and chemical of interest. PMID- 18507285 TI - Reduction of respirable silica following the introduction of water spray applications in Indian stone crusher mills. AB - Respirable crystalline silica dust generated during stone crushing operations has been linked to chronic lung disease and increased risk of tuberculosis. In India, most stone crushing mills operate without any dust control or containment systems. This investigation in the Khurda District of Orissa demonstrated a reduction in respirable particulate mass following the application of a fine mist of water. Average respirable quartz and cristobalite levels declined 82% and 69%, respectively, after water spray controls were installed. This finding suggests that relatively inexpensive modifications that are available in the local market can be effective at reducing silica exposures. Although average exposure levels, particularly during the dry season, may exceed the Permissible Exposure Limit for silica, the overall reductions observed were substantial. Widespread adoption of this simple control technology by stone crushers in India could have a positive public health impact. PMID- 18507286 TI - A survey of environmental and occupational work practices in the automotive refinishing industry of a developing country: Sonora, Mexico. AB - The automotive repair and refinishing industry has been studied intensively in industrialized countries, in part due to use of hazardous chemicals such as isocyanates and solvents, but little is known about industry practices in the developing world. The main objective of this paper was to investigate environmental and occupational work practices of this industry in a developing region, Sonora, Mexico. An integrated survey approach maximizes the opportunity for identifying risks as well as reducing risks. This investigation included detailed workplace visits to 41 body shops and 6 paint suppliers, as well as a survey of shop owners and 24 workers. Information was collected on work practices, level of technology in the shops, use of personal protective equipment, consumption and handling of hazardous chemicals and waste, hazard communication, and environmental consciousness. Most shops had little capital, outdated technology for exposure control, poor working conditions, high potential for exposure to hazardous chemicals, and little awareness of environmental and occupational health and safety. We concluded that work practices in the Sonoran auto refinishing industry are unsustainable and may pose a health risk to workers and the environment. PMID- 18507287 TI - Increased risk of death with congenital anomalies in the offspring of male semiconductor workers. AB - Female workers in the semiconductor industry have higher risks of subfertility and spontaneous abortion, but no studies exploring male-mediated developmental toxicity have been published. This study aimed to investigate whether the offspring of male workers employed in the semiconductor manufacturing industry had an increased risk of death with congenital anomalies. The 6,834 male workers had been employed in the eight semiconductor companies in Taiwan between 1980 and 1994. We identified the live born children with or without congenital anomalies of the workers using the National Birth and Death Registries from the Department of Health, Taiwan. Multiple logistic regression models were used to estimate the odds ratios (OR) of birth outcomes and deaths, controlling for infant sex, maternal age, and paternal education. A total of 5,702 children were born to male workers during the period 1980-1994. There were increased risks of deaths with congenital anomalies (adjusted OR, 3.26; and 95% confidence interval [CI], 1.12 9.44) and heart anomalies (OR, 4.15; 95% CI, 1.08-15.95) in the offspring of male workers who were employed during the two months before conception. We found evidence of a possible link between paternal preconception exposure of semiconductor manufacturing and an increased risk of congenital anomalies, especially of the heart. The possible etiological basis needs to be corroborated in further research. PMID- 18507288 TI - Occupational risks for idiopathic pulmonary fibrosis mortality in the United States. AB - Metal and wood dust exposures have been identified as possible occupational risk factors for idiopathic pulmonary fibrosis (IPF). We analyzed mortality data using ICD-10 code J84.1--"Other interstitial pulmonary diseases with fibrosis," derived age-adjusted mortality rates for 1999-2003, and assessed occupational risks for 1999, by calculating proportionate mortality ratios (PMRs) and mortality odds ratios (MORs) using a matched case-control approach. We identified 84,010 IPF deaths, with an age-adjusted mortality rate of 75.7 deaths/million. Mortality rates were highest among males, whites, and those aged 85 and older. Three industry categories with potential occupational exposures recognized as risk factors for IPF were identified: "Wood buildings and mobile homes" (PMR = 4.5, 95% confidence interval (CI) 1.2-11.6 and MOR = 5.3, 95% CI 1.2-23.8), "Metal mining" (PMR = 2.4, 95% CI 1.3-4.0 and MOR = 2.2, 95% CI 1.1-4.4), and "Fabricated structural metal products" (PMR = 1.9, 95% CI 1.1-3.1 and MOR = 1.7, 95% CI 1.0-3.1). Workers in these industry categories may benefit from toxicological studies and improved surveillance for this disease. PMID- 18507289 TI - Lung cancer mortality in a cohort of workers in a petrochemical plant: occupational or residential risk? AB - The purpose of the present study is to examine the role of environmental (non occupational) exposures in lung cancer risk among petrochemical workers at a large petrochemical plant built on the Sicilian coast in the immediate vicinity of the town of Gela, Italy in 1960. The cohort included workers employed in the Gela petrochemical plant in 1960-1993. We looked at mortality rates for the period 1960-2002. An internal comparison was performed between two categories of workers with different likelihood of residence in Gela during the period of employment. The rate ratio of mortality from lung cancer comparing "probable residents" with "possible non residents," adjusted for age, calendar period, andjob classification (only blue collar, only white collar and both), was 1.66 (90% Confidence Interval 1.07-2.58). Although the information collected is quite sparse and no inferences can be made about risk sources, the results show a possible excess of residential/environmental risk from lung cancer mortality for those workers more likely to have been residents in Gela. PMID- 18507290 TI - Determinants of health in seasonal migrants: coffee harvesters in Los Santos, Costa Rica. AB - In the agroexport zone of Los Santos Zone in Costa Rica, coffee is harvested by migrant labor. Most migrants are from Panama and Nicaragua. We describe migrants' housing- and service-related health determinants, with analyses of ethnicity, nationality and geography. We used interviews, observation-based assessments, and the Geographic Information System to assess a population of 8,783 seasonal migrants and 1,099 temporary dwellings at a total of 520 farms during 2004-2005. We identified determinants of poor health including widespread deficiencies in the quality of grower-provided dwellings, geographical isolation, crowding, lack of radio and television, and deficient toilets and cooking facilities. The indigenous and non-Costa Ricans shared the poorest conditions. Reluctance to use mainstream public health services was widespread, especially among foreign and indigenous migrants and the geographically isolated. Post-study, researchers organized workshops for audiences including workers, coffee producers, public officials and service providers. Topics have included migration, preventive health and hygiene, and child labor. This work was successful in convincing Costa Rican social security authorities to implement reforms that improve access to and quality of health care for the migrants. Special projects on ergonomics, psychosocial health hazards, and water quality, as well as a literacy program, are ongoing. PMID- 18507291 TI - Sex ratio changes as sentinel health events of endocrine disruption. AB - The production and widespread use of synthetic chemicals since the 1940s have resulted in ubiquitous contamination of fish, wildlife and human populations. Since the 1960s, observers have documented major damage to wildlife reproduction across the globe, and subsequently, damage to reproductive health in exposed humans as well. The sex ratio in human communities and populations can be readily measured to ascertain whether reproductive effects, such as subtle birth defects of the reproductive tract caused by exposures to chemicals, might be occurring. Male to female sex ratios appear to be declining in populations in several parts of the globe, possibly as a result of prenatal exposures to chemicals. Sex ratio data for communities with unusual occupational or environmental exposures can be compiled using traditional epidemiological techniques in pursuit of environmental justice. Local, regional and national population health researchers and occupational hygienists can use health statistics to examine sex ratios as sentinel health events that might portend patterns of subtle structural birth defects of the reproductive tract and functional deficits in neurodevelopment. PMID- 18507292 TI - Misuse of genomics in assigning causation in relation to benzene exposure. PMID- 18507293 TI - IJOEH and the critique of bias. PMID- 18507294 TI - The Institute of Occupational Medicine's role in semiconductor studies. PMID- 18507295 TI - Layperson's group remains concerned at IOM's role. PMID- 18507296 TI - AOEC patient rights and relationship to ACOEM. PMID- 18507297 TI - Clarification on mesothelioma rates in Ontario. PMID- 18507298 TI - IAOH denies industry influence. PMID- 18507299 TI - ASCLS continues collaborative efforts to address laboratory reimbursement and workforce issues. PMID- 18507300 TI - Practice levels and educational needs for clinical laboratory personnel. PMID- 18507301 TI - The doctorate in clinical laboratory science: CLS education beyond the baccalaureate. PMID- 18507302 TI - Strongyloidiasis: a review and update by case example. AB - A 77-year-old female immigrant from South America presented with epigastric pain, diarrhea, gastrointestinal bleeding, malabsorption, and acid reflux disorder. A gastroduodenoscopy, performed to assess for peptic ulcer disease, revealed parasitic larvae in the duodenal mucosa which were subsequently identified as Strongyloides stercoralis rhabditiform larvae. Anti-helminthic therapy was initiated to resolve infection. OBJECTIVES: Review the pathogenesis, diagnosis and treatment of strongyloidiasis; alert laboratory professionals to the importance of early detection of Strongyloides stercoralis in specimens from immigrants at risk and immunodeficient patients to reduce morbidity and mortality. PMID- 18507303 TI - Antimicrobial resistance of uncomplicated urinary tract infections in northern Utah. AB - OBJECTIVE: To evaluate antimicrobial resistance in uropathogenic bacteria in northern Utah. DESIGN: One hundred twenty bacterial isolates from community acquired UTI in the northern Utah area (Davis and Weber Counties) were tested. Samples were taken from otherwise healthy women, ages 18 to 50. Antimicrobial susceptibility testing for sulfamethoxazole/trimethoprim (SXT/TMP), ciprofloxacin, and nitrofurantoin comprised the process. SETTING: The Clinical Laboratory Science Department at Weber State University, with samples coming from clinics in the northern Utah area (Davis and Weber Counties). PARTICIPANTS: Urine samples were taken from otherwise healthy women, ages 18 to 50, who suffered from uncomplicated urinary tract infections. MAIN OUTCOME MEASURE: Antimicrobial resistance was measured using antimicrobial susceptibility testing and shown with other national resistance rates. RESULTS: Of bacterial isolates, 21.3% were resistant to SXT/TMP, 14.4% were resistant to ciprofloxacin, and 13.9% were resistant to nitrofurantoin. The resistance rates for ciprofloxacin and nitrofurantoin were acceptable for empirical UTI treatment (< 20% resistance), but local bacterial populations were found to demonstrate an increase in resistance to these two drugs as compared to previously observed national data. SXT/TMP resistance was above the recommended resistance threshold of 20% for effective empirical treatment, as advised by the IDSA. CONCLUSION: Results suggest that uncomplicated community-acquired UTI be treated with nitrofurantoin. Other recommendations include continued monitoring of local uropathogenic antimicrobial resistance. PMID- 18507304 TI - Evaluation of disinfectants on military NATO and DECON litters. AB - OBJECTIVE: This study evaluated the effectiveness of five disinfectants: A33, 10% bleach, 1% bleach, SPOROX, and 3% H2O2, on military NATO and DECON litters. DESIGN: Suspensions of Acinetobacter baumannii, Staphylococcus aureus, and spore enhanced Bacillus subtilis, with five percent albumin, were inoculated onto litters and dried overnight. The litters were saturated with disinfectant solutions and sampled after 10 minutes. The Log10 reduction in the number of bacteria recovered was calculated. SETTING: 59th Medical Wing, 59th Clinical Research Division, Lackland AFB TX. MAIN OUTCOME MEASURES: A reduction of > or =3 Log10 in the number of bacteria recovered from the test squares compared to the control squares was considered effective disinfection. RESULTS: On the NATO litter 10% bleach and SPOROX were effective against vegetative cells. On the DECON litter A33, 10% bleach, and SPOROX were effective against vegetative cells. After the 10 minute exposure none of the disinfectants evaluated were effective against spore-enhanced B. subtilis. CONCLUSION: When contaminated with vegetative cells military NATO and DECON litters can be effectively disinfected with a 10 minute exposure to some disinfectants. Further research is needed to find an effective disinfectant for spore contamination. PMID- 18507305 TI - The effect of ozone on common environmental fungi. AB - OBJECTIVE: To determine if gaseous ozone can effectively kill common environmental fungi. DESIGN: This study was designed to test the null hypothesis that there is no significant difference in viability between fungal conidia treated with ozone and fungal conidia not treated with ozone. A single control group design was utilized. SETTING: Academic research laboratory. INTERVENTIONS: Freshly prepared suspensions of Cladosporium spp., Stachybotrys spp., and Aspergillus niger conidia were diluted and plated onto the surface of solid agar plates. The plates were exposed to room air or to different concentrations of ozone for up to four hours, as were uninoculated plates. All plates were then incubated at 25 degrees C until quantitative colony counts could be performed. MAIN OUTCOME MEASURE: The effect of ozone on fungal conidia viability was assessed by comparing quantitative colony counts from conidia exposed to ozone to quantitative colony counts from conidia exposed only to room air. RESULTS: There was a significant (p < 0.05) decrease in viable conidia of all three fungi, at ozone concentrations of 5.0-12.8 parts per million, by four hours of exposure. However, in every case, some conidia remained viable even at the highest level of exposure. CONCLUSIONS: These data suggest that ozone must be used in conjunction with other methods of remediation or for more prolonged exposure times in order to eliminate fungal contamination of buildings. PMID- 18507306 TI - Bioethics--problems for today. PMID- 18507307 TI - Case one: Patient autonomy and the freedom to act against one's self-interest. AB - A 16-year-old Hodgkin lymphoma patient refuses to have his blood specimen drawn, thus canceling his scheduled oncologic treatment. As a 16-year-old, he has no legal standing as an adult. His parents are split over his decision. One supports his right to choose; the other wishes the specimen to be drawn and the chemotherapy reinstated. The physicians at the hospital are seeking legal redress to have the court order the blood specimens to be taken. PMID- 18507308 TI - Case one: Patient interests and medical paternalism. AB - A 16-year-old Hodgkin lymphoma patient refuses to have his blood specimen drawn, thus canceling his scheduled oncologic treatment. As a 16-year-old, he has no legal standing as an adult. His parents are split over his decision. One supports his right to choose; the other wishes the specimen to be drawn and the chemotherapy reinstated. The physicians at the hospital are seeking legal redress to have the court order the blood specimens to be taken. PMID- 18507309 TI - Case two: Experimental blood substitute on first response vehicles. AB - Clinical trials in a number of countries are now underway to evaluate experimental, non-human blood substitute. One scenario calls for the blood substitute to be available on board emergency vehicles. This allows first responders the opportunity to provide transfusion support at an accident site and on the way to the hospital. However, many of the patients who would most benefit from the use of this material may be unconscious and unable to comprehend or sign an informed consent. One possible solution would be to eliminate the need for informed consent. PMID- 18507310 TI - Case two: A Kantian approach to the morality of blood substitute clinical trials without informed consent. AB - Clinical trials in a number of countries are now underway to evaluate experimental, non-human blood substitute. One scenario calls for the blood substitute to be available on board emergency vehicles. This allows first responders the opportunity to provide transfusion support at an accident site and on the way to the hospital. However, many of the patients who would most benefit from the use of this material may be unconscious and unable to comprehend or sign an informed consent. One possible solution would be to eliminate the need for informed consent. PMID- 18507311 TI - Case three: Ethics of coercion. AB - A woman is murdered in a small town. At autopsy, the pathologist notes the woman had engaged in sexual relations shortly before her murder. The police department determines the male partner should be considered a person of interest in their investigation. They begin a canvas of the town, asking every male to voluntarily consent to a DNA test. Men refusing to provide the specimen will be publicly listed as potential suspects and perhaps arrested. All 1500 men in the town provide a specimen and none is identified as the sex partner. The DNA results are entered into the FBI's database and made available to every law enforcement agency in the country. PMID- 18507312 TI - Case three: Collection of evidence in a murder investigation. AB - A woman is murdered in a small town. At autopsy, the pathologist notes the woman had engaged in sexual relations shortly before her murder. The police department determines the male partner should be considered a person of interest in their investigation. They begin a canvas of the town, asking every male to voluntarily consent to a DNA test. Men refusing to provide the specimen will be publicly listed as potential suspects and perhaps arrested. All 1500 men in the town provide a specimen and none is identified as the sex partner. The DNA results are entered into the FBI's database and made available to every law enforcement agency in the country. PMID- 18507313 TI - Linguistic boundaries as predictors of the time between letters in oral and typed spellings. AB - The authors investigated the importance of boundaries between phonemes, syllables, onsets and rimes, and morphemes in English spelling. They analyzed oral spelling data from a previous sample of 17 college students to predict time between consecutive letters (pause time) on the basis of the presence or absence of each linguistic boundary. The authors used a parallel approach to analyze pause times of 30 college students when typing individual words and when typing words in an essay. For oral and typed spellings of individual words, syllable boundaries significantly predicted pause times. Phoneme boundaries also predicted pause times in typed spellings of individual words. For typing essays, only onset rime boundaries significantly predicted pause times. The results support the importance of syllables in the spelling of individual words. Further, the results suggest that spelling in the context of writing is a qualitatively different process than is spelling individual words by dictation. PMID- 18507314 TI - An investigation of time course of category and semantic priming. AB - Low semantically similar exemplars in a category demonstrate the category-priming effect through priming of the category (i.e., exemplar-category-exemplar), whereas high semantically similar exemplars in the same category demonstrate the semantic-priming effect (i.e., direct activation of one high semantically similar exemplar by another). The author asked whether the category- and semantic-priming effects are based on a common memory process. She examined this question by testing the time courses of category- and semantic-priming effects. She tested participants on either category- or semantic-priming paradigm at 2 different time intervals (6 min and 42 min) by using a lexical decision task using exemplars from categories. Results showed that the time course of category priming was different from that of semantic priming. The author concludes that these 2 priming effects are based on 2 separate memory processes. PMID- 18507315 TI - The good-subject effect: investigating participant demand characteristics. AB - Although researchers are often concerned with the presence of participant demand, few have directly examined effects of demand on participant behavior. Before beginning the present study, a confederate informed participants (N = 100) of the study's purported hypothesis. Participants then performed a laboratory task designed to evaluate the extent to which they would respond in ways that may confirm or disconfirm the hypothesis of the study. The authors found that participants tended to respond in ways that confirmed the hypothesis, yet this tendency depended on attitudes toward the experiment or experimenter and other individual differences. In addition, results suggested that suspicion probes may be ineffective in measuring participants' previous knowledge and suspicion. Findings indicate the need for more research and consideration of demand in the design of studies and analysis of data. PMID- 18507316 TI - Precuing an isolated stimulus temporarily outweighs in-stream stimulus facilitation. AB - The flash-lag effect (FLE), when studied in rapid serial visual presentations, has shown that participants perceived a target presented in a stream prior to its simultaneous out-of-stream replica (T. Bachmann & E. Poder, 2001). In the present study, the authors introduced a precue to the single flashed, isolated target to see if drawing bottom-up attention to the flash also would compensate for the effects of the mechanisms that influence the processing of in-stream items. The extent of the flash-lead effect in the precued condition was considerably larger than the lead-effect in earlier experiments and in a control experiment where no precue was presented for the flashed targets. For the first 200-250 ms from the beginning of the stream, flash-lead dominated. The FLE did not appear until 400 ms after stream onset. The authors interpret results as the combined relative effects of in-stream target masking, in-stream sensory facilitation, and spatial attentional facilitation. PMID- 18507317 TI - Perceived freedom-responsibility covariation among Cypriot adolescents. AB - Participants were 67 Cypriot adolescents who responded to propositions regarding positive, negative, and noncontingent relations between freedom and responsibility. The authors framed items so that half dealt with freedom given responsibility, and the other half dealt with responsibility given freedom. Results indicated participants were more likely to endorse positive-contingency items than they were negative and noncontingency items when items were framed around freedom given responsibility. However, when items were framed around responsibility given freedom, no such differences emerged. The authors discuss results relative to cultural and sociopolitical differences and similarities between children in Cypress and participants in the United States and implications concerning the present study and previous studies regarding these constructs. PMID- 18507318 TI - Effects of conceptually based familiarity in memory conjunction errors. AB - The authors examined whether the frequency of conjunction errors varied in a recognition test according to the semantic relation between 2 words. Participants studied a series of natural (meaningful) or bizarre Japanese noun phrase patterns and later completed a recognition test that contained old, conjunction, feature, and new stimuli. Participants who studied a list of natural noun phrases did not make any false old responses to unmatched (bizarre) conjunction stimuli. However, participants who studied bizarre noun phrases made as many false old responses to matched (bizarre) conjunction stimuli as to unmatched (natural) conjunction stimuli. The results of the bizarre study stimuli group indicated that bizarre noun phrases may be translated into meaningful stimuli because of mnemonic instruction. Together, the results support the hypothesis that conceptually based familiarity, not pure perceptually based familiarity, influences false old responses to conjunction stimuli. PMID- 18507319 TI - Concierge medicine: legal issues, ethical dilemmas, and policy challenges. AB - The practice of "concierge" or "retainer" medicine is growingly steadily due to economic and legal pressures on physicians. This practice model, which typically involves charging access or subscription fees to a limited pool of patients, raises legal hazards, contractual challenges, and ethical dilemmas for physicians interested in converting to concierge medicine, as well as important health policy questions. This article examines these legal and contractual issues, and discusses some of the ethical and policy implications of this relatively new form of medical practice. The authors conclude that this innovative practice form provides a beneficial alternative for patients seeking more personal, proactive, and/or intensive medical care. As concierge medicine proliferates, a wider range of practice models and fee structures should make this approach more affordable to lower and middle income patients--and help satisfy the need for greater access to preventive healthcare. Health policy makers should take steps to encourage this outcome. PMID- 18507320 TI - Can preventing diversion of profitable patients justify hospitals' economic credentialing under the antitrust laws? AB - Investments by physicians in specialty hospitals, ambulatory surgery centers, and other competing facilities have caused many full-service acute care hospitals a great deal of concern. These full-service hospitals believe they are at a disadvantage in competing against such centers because physician-investors allegedly can "pre-select" profitable patients. Full-service hospitals have reacted in a variety of ways, including adopting economic credentialing policies. Physician-owned centers often view these policies as anti-competitive and seek relief under the antitrust laws. Hospitals attempt to defend these actions on the grounds that they prevent what economists call "free-riding." A key element of this defense is whether pre-selection is occurring and whether the hospital has sufficient evidence of pre-selection to justify the conduct under the antitrust laws. PMID- 18507321 TI - HIPAA and ex parte interviews--the beginning of the end? AB - For many legal counsel, ex parte interviews with a treating physician have served as a time-honored method of informal discovery. In particular, litigators have used ex parte interviews with a former or current physician to obtain personal health information about a party or witness. Both before and after the Health Insurance Portability and Accountability Act of 1996 (HIPAA) Privacy Rule, privacy concerns developed regarding ex parte interviews. Has HIPAA's Privacy Rule initiated the beginning of the end for informally obtaining health information? This article examines ex parte interviews before the HIPAA Privacy Rule; how courts have addressed ex parte interviews after the Privacy Rule; conflicting issues physicians face in deciding whether to participate in ex parte interviews; and options for legal counsel to consider. PMID- 18507322 TI - Home healthcare workers and the Fair Labor Standards Act. AB - In a major industry shift, long term care is moving from nursing homes and institutions to the private home. This change results from a number of factors, including the lower cost of home-based care. These lower costs can be traced to a number of exemptions from the Fair Labor Standards Act requirements for minimum wage and premium overtime payments which apply to many home healthcare workers. These include the companionship, live-in, and professional exemptions. As the home healthcare industry has grown, home healthcare workers have challenged the applicability of these exemptions. This article will explore the issues reflected in those challenges and their resolution, and provide suggestions to help employers ensure that their employees fall within the exemptions. PMID- 18507323 TI - Drafting arbitration provisions for complex business litigation in healthcare. PMID- 18507324 TI - Perinatal loss: a qualitative study in Northern Ireland. AB - This article describes the experiences of women in Northern Ireland who have experienced a miscarriage or stillbirth. Pregnancy loss encompasses several dimensions of loss for women, loss of the future, loss of self-identity, and the loss of anticipated parenthood. The study explored how women emotionally responded to loss and the care they received from medical staff. Burial arrangements for the remains of the baby are also explored. The methodology adopted a narrative approach based upon in-depth interviews with 23 women who attended pregnancy loss self-help groups. The women's narratives highlight their emotional responses to loss, the medicalization of perinatal grief, and burial arrangements. Women felt that their experience was emotionally negative in that they had been subjected to a rationalizing process of medicalization. The primary focus for the women was on the need to recover space for their emotions and seek acceptance and recognition of the validity of their grief. The study demonstrated that the women's response to being marginalized led them to make sense of their experiences and to create spaces of resistance to medicalization. The way in which women placed emotion at the center of their narratives is taken to be a powerful indicator that the support they require from professionals should take account of the meanings they have constructed from their experience of loss. PMID- 18507325 TI - The social relations of bereavement in the Caribbean. AB - The objectives of this article are to discuss the various types of behaviors associated with grief and bereavement, and to examine the relationships, consequences, and outcomes of bereavement practices among the various religious and ethnic groups in the English-speaking Caribbean Islands of Jamaica, Trinidad, Grenada, and Barbados. The rituals associated with death and grief differs across cultures and is greatly influenced by religious beliefs and traditions. How these rituals are played out depend on the culture of origin and level of acculturation of the various groups into mainstream society. In the Caribbean region, expressions of grief represent religious and cultural traditions that may have a significant impact on social relations, particularly in multi-ethnic and multicultural societies. In the English-speaking Caribbean Islands of Jamaica, Trinidad, Grenada, and Barbados, mourning follows the patterns of traditional religious practices which have remained consistent over time. While families and friends may offer social support before and after burial or cremation, the social aspects of bereavement may also have implications for inter-group relations. Insights into bereavement practices and what it holds for ethnic and religious groups in contemporary Caribbean are presented. PMID- 18507326 TI - The death of mourning: from Victorian crepe to the little black dress. AB - Mourning is a natural response to loss. In the late eighteenth century and throughout the nineteenth century, in England and France, the bereaved was expected to follow a complex set of rules, particularly among the upper classes, with women more bound to adhere to these customs than men. Such customs involved wearing heavy, concealing, black costume and the use of black crepe veils. Special black caps and bonnets were worn with these ensembles. Widows were expected to wear these clothes up to four years after their loss to show their grief. Jewelry often made of dark black jet or the hair of the deceased was used. To remove the costume earlier was thought disrespectful to the deceased. Formal mourning culminated during the reign of Queen Victoria. Her prolonged grief over the death of her husband, Prince Albert, had much to do with the practice. During the succeeding Edwardian rule, the fashions began to be more functional and less restrictive, but the dress protocol for men and women, including that for the period of mourning, was still rigidly adhered to. When World War I began, many women joined the workforce. Most widows attempted to maintain the traditional conventions of mourning, but with an increase in the number of casualties, it became impractical for them to interrupt their work in order to observe the seclusion called for by formal mourning etiquette. Never had the code of mourning been less strictly applied than during this period. The mourning outfits of the time were modest and made of practical materials. Little jewelry and few other accessories were used. Certain aspects of traditional mourning were still followed, such as the use of jet beading, crepe trim, and widows' caps. However, the hemlines fell above the ankle, the veil was used to frame the face instead of cover it, and the v-neckline left the chest and neck bare. During the following decades, gradually the rules were relaxed further and it became acceptable for both sexes to dress in dark colors for up to a year after a death in the family. PMID- 18507327 TI - "The wholeness of things": infusing diversity and social justice into death education. AB - This article presents resources and strategies for the infusion of diversity and social justice themes into an undergraduate death and dying course. The intent is not to replace or dismiss existing thanatological insights and debates, but rather to widen the cultural perspective to bring these insights and debates into conversation with multiple ways of perceiving and understanding. The article covers definitions, goals and rationales, challenges in identifying and developing appropriate resources, and overall course design. It also explores readings, audiovisual materials, class activities, and pedagogical approaches to foster: student engagement with diverse world-views and experiences; understanding of "cultural competence" in various fields; awareness of the impact of race, class, gender, etc., on access to resources and care; commitment to activism for social justice; and exposure to many forms of resilience, meaning making, and creative healing. PMID- 18507328 TI - Turkish health professional's attitude toward euthanasia. AB - The cross-sectional study was administrated between April and September 2006. Participants are doctors, nurses, and midwives. Between these dates we met only 750 health staff (doctor, nurse, and midwife). Six hundred thirty-two of them responded to our questionnaire, 122 of them were in Manisa city, and 510 of them in Erciyes. We sought to identify variables that contribute to euthanasia attitude, including demographics, in order to demonstrate Turkish doctors', nurses', and midwives' attitudes toward euthanasia and to compare their attitudes in this regard. The data was collected by a two-part questionnaire. The first part included questions about the health personnel; the second part comprised the euthanasia (Medical Staffs Attitude toward Euthanasia) scale. The scale was developed by the researcher to measure the attitude of healthy staff euthanasia. The SPSS was used to analyze the data. Student t-test, ANOVA, Mann Whitney U, and Kruskal Wallis were used to evaluate the data. Thep value 0.05 (95% confidence interval) was accepted as significant. In our study, professional groups are compared with all the factors but there is a significant difference only between social cost and professional groups. PMID- 18507329 TI - Does religiosity help Muslims adjust to death?: a research note. AB - Death is the end of life. But Muslims believe death is an event between two lives, not an absolute cessation of life. Thus religiosity may influence Muslims differently about death. To explore the impact of religious perception, thus religiosity, a cross-sectional, descriptive, analytic and correlational study was conducted on 150 Muslims. Self-declared healthy Muslims equally from both sexes (N = 150, Age range--20 to 50 years, Minimum education--Bachelor) were selected by stratified sampling and randomly under each stratum. Subjects, divided in five levels of religiosity, were assessed and scored for the presence of maladjustment symptoms and stage of adjustment with death. ANOVA and correlation coefficient was applied on the sets of data collected. All statistical tests were done at the level of 95% confidence (P < 0.05). Final results were higher than the table values used for ANOVA and correlation coefficient yielded P values of < 0.05, < 0.01, and < 0.001. Religiosity as a criterion of Muslims influenced the quality of adjustment with death positively. So we hypothesized that religiosity may help Muslims adjust to death. PMID- 18507330 TI - Cultural values, life experiences, and wisdom. AB - Wisdom is considered one ideal endpoint of human development across cultures. Studies have provided evidence for certain facilitating conditions such as challenging and stressful life events because they increase differentiation through accommodative changes, resulting in greater tolerance for uncertainty, and less projection tendencies and self-centeredness. Positive experiences may also facilitate wisdom by fostering integration and coherence. However, cultural values, particularly conservation and openness, may moderate these experiences for older adults. In a sample of middle-aged to older community dwelling European American adults (n = 97), results suggested that experiencing a macrosocial event as a negative experience, and spiritual/existential as a positive experience, interacted with conservation value to predict transcendent wisdom. Among Vietnamese American adults (n = 102), macrosocial event alone was negatively related to transcendent wisdom. These results suggest that not endorsing conservation value as one grows older and experiences different life events is beneficial for wisdom. PMID- 18507331 TI - Age-related differences in worry and related processes. AB - This study investigated the hypothesis that older adults would show age-related reductions in the tendency to worry in both their retrospective accounts and through cross-sectional age comparisons with a sample of younger adults. We also sought to determine whether age differences would be evident in psychological processes associated with a tendency to worry in general adult samples (intolerance of uncertainty and beliefs in the functional value of worry). Support was found for the hypothesized age-related reduction in worry in both retrospective reports among the older adults and cross-sectional age comparisons. Older adults were also found to report less intolerance of uncertainty and less belief in the functional value of worrying compared to younger adults. These findings suggest that worry-proneness is reduced in late adulthood and that a greater ability to tolerate uncertainty in life and to see less value in worrying may partially account for this. Developmental and contextual changes that may account for these findings are considered. PMID- 18507332 TI - Ravishing or ravaged: women's relationships with women in the context of aging and Western beauty culture. AB - We undertook this narrative analysis study to explore the complexities of women's relationships with other women within the sociocultural milieu of beautyism and ageism. Using an open-ended narrative framework, four focus groups of women living in different regions throughout the U.S. were conducted and analyzed to identify thematic categories within and across group sessions. We discuss four of the six key themes discovered in response to the primary research question: How does beauty culture shape women's experiences of aging and their relationships with women of all ages? We conclude that Western idealized beauty standards exert a divisive impact on women's relationships with each other across the life span, negatively affecting their socioemotional well-being, especially in old age. PMID- 18507333 TI - The relation between self-determination and retirement satisfaction among active retired individuals. AB - Little is known about the motivational forces leading retired individuals to engage in post-retirement activities and how they could be related to satisfaction with retirement. Using the self-determination framework, the purpose of the present exploratory study was to examine the nature of active retirees' motivation and its impact on satisfaction with life in retirement. The Global Motivation Scale and measures of satisfaction with retirement were administered to 150 active retired individuals engaged in organized post-retirement activities in a University-based organization. Results revealed that these individuals were mainly characterized by higher levels of intrinsic motivation for knowledge, stimulation and accomplishment, rather than extrinsic dimensions. Regression analysis further demonstrated that intrinsic motivation for both accomplishment and stimulation were positively related to satisfaction with retirement, over and above the significant contribution of time since retirement, anticipated satisfaction with retirement, and subjective health. This exploratory study highlights the motivational mechanisms through which post-retirement behaviors could positively influence satisfaction with life in retirement among active retirees. PMID- 18507334 TI - [Advances in the study of molecular mechanisms, applications and screening for altered peptide ligand]. AB - Altered peptide ligand (APL), a short peptide with immune regulatory activity and substitutions of a single or multiple amino acids in an antigenic peptide, has shown potential therapeutic effect on autoimmune disease, tumor and virus infection. APL regulates immune responses by interfering the interaction between the major histocompatibility complex (MHC), antigenic peptide and T cell receptor (TCR), or by regulating the intracellular signaling of antigen presenting cells, bystander suppression and inducing heterogenous immune responses. High-specific and high-affinity APL screened from peptide laboratory by phage display, has a potential to be a new resource for drug with antigen specificity. PMID- 18507335 TI - [Process of HIV-1 reverse transcription and its detection by using PCR]. AB - Human immunodeficiency virus (HIV) is a retrovirus, belongs to Lentiviridae family. As long as viral genetic material entering into host cytoplasm, double strand DNAs synthesis occurs which is catalyzed by reverse transcriptase (RT) with viral plus-strand RNA as template. This reverse transcription is a key link of HIV-1 life cycle and an important target for anti-HIV drug development. The process of reverse transcription can be divided into several steps: formation of minus-strand strong-stop DNA; the first translocation; initiation of plus-strand DNA synthesis; and, the second translocation and the completion of both strands. These steps can be detected individually by using polymerase chain reaction (PCR) according to the amplified products on the region of R/U5, U3, U5/PBS and the sequence between LTR and Gag. In this review, we summarize the principle for detecting stages of HIV-1 reverse transcription by using PCR. PMID- 18507336 TI - [Progress in the study of pH and temperature sensitive biodegradable block copolymers]. AB - pH and temperature sensitive biodegradable block copolymers are some macromolecules connected by biodegradable materials and pH sensitive monomers according to a certain sequence, or biodegradable polyesters polymerized themselves. On the basis of pertinent documents, the development of pH and temperature sensitive biodegradable block copolymers was introduced, involving their mechanism of action and potential application. PH and temperature sensitive biodegradable block copolymers could control the drug release rate freely, avoiding burst effect. Besides, the biocompatibility of these biodegradable materials is also excellent. So the use of pH and temperature sensitive biodegradable block copolymers as biodegradable drug delivery devices has attracted considerable interest in the intelligent drug delivery system. PMID- 18507337 TI - [Effect of resveratrol on chronic obstructive pulmonary disease in rats and its mechanism]. AB - The purpose of this study is to establish COPD animal model by intra-tracheal instillation of bleomycin (BLM) once and exposure to cigarette smoke for continuous 27 d, and to observe the effects of the inhalation on the model. At the 29th day, blood samples were taken from cervical artery for blood-gas analysis and parameters of lung function were recorded. Bronchoalveolar lavage fluid (BALF) was collected to measure intercellular adhesion molecule-1 (ICAM-1) concentration. The results showed that atomization inhaled resveratrol could alleviate rat COPD lung injury accompanied by amelioration of pathological changes, increase the ratio of forced expiratory volume in 0.3 s (FEV0.3) and forced vital capacity (FVC), and decrease the ICAM-1 level in BALF. The ultimate reduction of inflammatory factors was involved, at least in part, in the mechanism of resveratrol effects. PMID- 18507338 TI - [Effect of nordy on the function of endothelial progenitor cells from human umbilical cord blood induced by vascular endothelial growth factor]. AB - This study is to investigate whether the synthesized chiral compound Nordy has influence on the function of endothelial progenitor cells (EPCs) from human umbilical cord blood induced by vascular endothelial growth factor (VEGF). EPCs were isolated from human umbilical cord blood by density gradient centrifugation. After cultured for 7 -10 days, EPCs were prepared for detecting effect of Nordy on proliferation, migration and tubule-forming activity in Matrigel induced by VEGF. Incubation of EPCs with 100 micromol L(-1) Nordy for 24 h initially inhibited the proliferative capacity of EPCs induced by VEGF (P <0.05). Moreover, 25 -50 micromol L(-1) Nordy also exhibited inhibitory effect at 48 -72 h. In addition, 25 - 100 micromol L(-1) Nordy impaired EPCs migratory and tubule forming capacity in vitro (P < 0.05). Nordy could inhibit in EPCs the functions of proliferation, migration and tubulogenesis induced by VEGF in vitro, which might be a possible mechanism of its anti-EPCs effects. PMID- 18507339 TI - [Induction of G2 /M phase arrest and apoptosis of MCF-7 cells by novel benzofuran lignan via suppressing cell cycle proteins]. AB - In the present study, a newly synthesized benzofuran lignan 4-formyl-2-(4-hydroxy 3methoxyphenyl)-5-(2-methoxycarbonyethyl)-7-methoxy-benzo [b] furan (ERJT-12) was tested for its antiproliferative activity on human tumor cells. The related mechanisms were also investigated. In vitro growth inhibitory effects of ERJT-12 on various cancer cell lines were determined by MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometry. The integrity of DNA was assessed by agarose gel electrophoresis. Activation of Caspase-3/7 and Caspase-6 was measured by colorimetric assay. The expressions of cell cycle proteins cell divide cycle 25c (Cdc25c), cyclin dependent kinase 1 (CDK1), CyclinB1 and apoptosis-related proteins Bax and Bcl-2 were detected by Western blotting. MTT assay showed that ERJT-12 inhibited the proliferation of several cancer cell lines including multidrug resistant cells. MCF-7 cells were markedly arrested at gap2/mitosis (G2/M) phase after treatment with ERJT-12 and progressed into apoptosis. The increased activities of Caspase-3/7 and Caspase-6 in MCF-7 cells were observed. The expression of CyclinB1 was down-regulated. The activities of Cdc25c and CDK1 protein were suppressed and Bcl-2 protein was phosphorylated. ERJT-12 displays potent antiproliferative activity towards cancer cells through suppressing cell cycle proteins, arresting cell cycle at G2/M phase and inducing apoptosis. It might be a novel candidate for cancer therapy. PMID- 18507340 TI - [Hypocholesterolemic effect of stilbene extract from Cajanus cajan L. on serum and hepatic lipid in diet-induced hyperlipidemic mice]. AB - Cajanus cajan L. is a natural plant, which contains a lot of potential active components. In the present study, we identified the effects of the stilbene extract from Cajanus cajan L. (sECC) on hepatic cholesterol metabolism in diet induced (for 4 weeks) hyperlipidemic Kunming mice. All experimental mice were divided into 5 groups: control group, high lipid model group, sECC-treated with 200 or 100 mg kg(-1), and simvastatin (Sim, 12 mg kg(-1)) treated group. The mice were fed with fat and cholesterol-enriched chow except control mice that were fed with standard diet. The effects of sECC were investigated by monitoring serum and liver lipid profile (i. e. cholesterol homeostasis) in mice. To further explore the mechanism of sECC, hepatic cholesterol 7alpha-hydroxylase (CYP7A1) and low density lipoprotein (LDL) receptor expressions in cholesterol homeostasis were analyzed by reverse transcription PCR. After 4 weeks pretreatment, the mice in the high lipid model group showed markedly higher serum and hepatic lipid contents than control group (P< 0.01). Compared with high lipid model group, the increased serum and hepatic lipid contents were markedly attenuated by sECC (200 mg kg(-1)), the serum and hepatic total cholesterol were reduced by 31.5% and 22.7% (P<0.05), respectively. The triglyceride contents of serum and liver were also lowered by 23.0% and 14.4%, respectively. At the same times, serum LDL cholesterol decreased by 53.0% (P<0.01). The mRNA expressions of hepatic CYP7A1 and LDL-receptor were significantly enhanced in the mice administered with sECC (200 mg kg(-1)), whereas those expressions were suppressed by the fat and cholesterol-enriched diet. These data indicate that sECC reduces the atherogenic properties of dietary cholesterol in mice. It is indicated that expression enhancement of hepatic LDL-receptor and cholesterol 7alpha-hydroxylase may be responsible for the hypercholesterolemic effect. PMID- 18507341 TI - [Protective effect of urantide against myocardial ischemia injury]. AB - This study is to investigate the protective effect of urantide against myocardial ischemia injury in mice and its mechanism. The ischemic model was made by using subcutaneous injection of isoproterenol (Iso) in mice, the change of ST segment of electrocardiogram (ECG) was observed, and the activitise of lactate dehydrogenase (LDH) and nitric oxide synthetase (NOS), the contents of malonaldehyde (MDA) and nitric oxide (NO) in serum were measured. The histopathological changes of myocardium were observed by using HE staining. The anoxia/reoxygenation (A/R) model of myocardial cells on neonatal Sprague-Dawley rats was established. Methyl thiazolyl tetrazolium (MTT) assay and confocal microscopy were respectively used to measure the viability and intracellular Ca2+ concentration in myocardial cells exposed to A/R. LDH activity and cTnI content in the cell culture medium were assayed for the evaluation of myocardial cells injury. The results revealed that urantide in the range of 3 - 30 microg kg(-1) iv markedly inhibited Iso-induced raise of the ST segment of ECG; 10 and 30 microg kg(-1) significantly reduced the increases of MDA content and LDH activity in mice serum, remarkably raised the activity of NOS and the content of NO. Urantide (10 and 30 microg kg(-1)) also significantly ameliorated myocardial ischemic injury. On the A/R model of myocardial cells, urantide (1 x 10(-6) - 1 x 10(-9) mol L(-1)) could evidently inhibit the increases of cTnI content, reduce the rise of intracellular Ca2+ concentration. Urantide (1 x 10(-6) - 1 x 10(-7)) mol L(-1) increased the viability of myocardial cells injured by A/R and cut down LDH activity in the cell culture medium. Therefore urantide has significant protective effect against myocardial ischemia or A/R injury via the inhibition of Ca2+ overload and the augmentation of NO synthesis. PMID- 18507342 TI - [Synthesis and anticancer activity of norcantharidin-galactose derivatives]. AB - To design and synthesize the hepatic targeting anticancer prodrug with norcantharidin (NCTD-Gal) conjugating structure, galactosylated NCTD derivatives were synthesized from NCTD analogues modified by a series of amino acids via acylation, hydrolysis, glycosylation and deacetylation. Seven new compounds were synthesized as beta-O-glycosides and characterized by IR, MS, NMR and element analysis. The compound 4a was chosen for the inchoate antitumor experiments on mice. The result showed that the antitumor inhibition rate of 4a groups with medium and high dose are clearly higher than that of NCTD group, which suggests that anticancer effect of NCTD is improved at a certain degree by galactosylation. PMID- 18507343 TI - [Synthesis and biological activity of a series of benzopyran derivatives]. AB - Thirteen benzopyran derivatives were synthesized and their activity stimulating the differentiation of preadipocytes into adipocytes were evaluated with 3T3-L1 cells. Compound 8 was also tested for its hypoglycemic activity on db diabetes mice model. Results indicated that compounds 3, 8 and 11 exhibited strong differentiation-stimulating activity on 3T3-L1 cells model, and compound 8 can reduce the blood-sugar level of db diabetes mice dramatically. PMID- 18507344 TI - [A novel triterpenoid saponin from Prunella vulgaris]. AB - To study the constituents of the Prunella vulgaris L, the constituents were isolated by various column chromatography and the structures were identified on the basis of chemical and spectral analysis. One saponin compound (I) and one flavone glycoside compound (II) were obtained from Prunella vulgaris L. Their structures were elucidated as 16-oxo-17-demethyl-3beta,24-dihydroxylolean-12-en-3 O-beta-D-glucuronoside (I), and acacetin-7-O-beta-D-glucopyranoside (II). Compound I is a novel triterpenoid saponin and named as prunelloside A. Compound II was obtained for the first time from the Prunella genus. PMID- 18507345 TI - Chemical constituents from the leaves of Broussonetia papyrifera. AB - To separate and identify the chemical constituents from the leaves of Broussonetia papyrifera (Linn.) Vent, various columns including Diaion HP-20, Toyopearl HW-40C, Sephadex LH-20, silica gel were employed for the isolation and purification of compounds from the leaves of B. papyrifera. The structures of the compounds were elucidated by their physiochemical characteristics and spectral data. Nineteen compounds were isolated from the leaves of B. papyrifera and their structures were identified as apigenin (1), apigenin-7-O-beta-D-glucopyranoside (2), chrysoerid-7-O-beta-D-glucopyranoside (3), apigenin-7-O-beta-D glucopyranuronide (4), vitexin-7-O-beta-D-glucopyranoside (5), luteolin (6), 5,7,4'-trihydroxyl-6-C-[a-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranosyl flavone (7), 5,7,4'-trihydroxyl-8-C-[a-L-rhamnopyranosyl (1-->2)]-beta-D glucopyranosyl flavone (8), saponaretin (9), vitexin (10), benzyl benzoate-2, 6 di-O-beta-D-glucopyranoside (11), (2R, 3R, 5R, 6S, 9R)-3-hydroxy-5,6-epoxy-beta ionol-2-O-beta-D-glucopyranoside (12), (2R, 3R, 5R, 6S, 9R)-3-hydroxyl-5,6-epoxy acetyl-beta-ionol-2-O-beta-D-glucopyranoside (13), ficustriol (14), (6S, 9S) roseoside (15), 3beta-hydroxy-5alpha,6alpha-epoxy-beta-ionone-2alpha-O-beta-D glucopyranoside (16), icariside B1 (17), sammangaoside A (18), 3-hydroxy 5alpha,6alpha-epoxy-beta-ionone (19). Compounds 11, 12 and 13 are new compounds, the others are isolated from this genus Broussonetia for the first time. PMID- 18507346 TI - [Chemical constituents from the tuber of Cremastra appendiculata]. AB - To study the chemical constituents of "Shan-Ci-Gu" (the tuber of Cremastra appendiculata (D. Don) Makino), the compounds were isolated with silica gel and reverse phase silica gel as well as Sephadex column chromatographic method. Their structures were elucidated on the basis of modern spectra technology. Seven compounds were isolated and identified as 5-methoxybibenzyl-3, 3'-di-O-beta-D glucopyranoside (1), militarine (2), loroglossin (3), protocatechuic acid (4), succinic acid (5), gastrodin (6), and daucosterol (7). Compound 1 is a new compound. Compounds 2 -6 were isolated from this plant for the first time. PMID- 18507347 TI - Comparison of the profiles of chlorogenic acids and their derivatives from three Chinese traditional herbs by LC-MSn. AB - To investigate the chlorogenic acids resources in Eucommia ulmoides Olive leaves, Lonicera japonica Thunb. leaves and Houttuynla cordata Thunb. leaves, methanolic extracts of these three materials have been analysed qualitatively for chlorogenic acids and their derivatives by structure-diagnostic LC-MSn. Three monocaffeoylquinic acids (3-CQA, 4-CQA, 5-CQA) were detected. 5-CQA dominated this subgroup in Eucommia ulmoides Olive and Lonicera japonica Thunb. leaves, but 3-CQA and 4-CQA dominated this subgroup of chlorogenic acids in the leaves of Houttuynla cordata Thunb. Caffeoylquinic acid-glycosides were detected for the first time from Eucommia ulmoides leaves. 5-FQA was found in Lonicera japonica Thunb. leaves, and 3-FQA and 4-pCoQA have been identified in Houttuynla cordata Thunb. This is the first report of the chlorogenic acid profile in Houttuynla cordata Thunb. The comparatively unusual profile of caffeoylquinic acids in Houttuynla cordata Thunb. makes it a convenient source of 3-CQA and 4-CQA that are not commercially available. PMID- 18507348 TI - [Studies on change of chemical composition in coptis-scute herb couple by using HPCE and HPLC]. AB - Quantitative analysis on different proportions of couple of Coptis and Scute. The proportions of Coptis and Scute couple were 6:1, 6:4, 6:7, 6:10, 6:13, separately. The contents of alkaloids in Coptis and decoction of Coptis and Scute were determined by HPCE and the contents of flavones in Scute and decoction of Coptis and Scute were determined by HPLC, separately. Precipitates were generated after the couple of Coptis and Scute, which results the decrease of contents of effective ingredients. PMID- 18507349 TI - [A new qualitative and quantitative analytical method of chromatographic fingerprints: total quantum statistical moment]. AB - A new qualitative and quantitative analytical method of the chromatographic fingerprints: the Total Quantum Statistical Moment (TQSM) has been eluciated and established according to statistical moment principle. The study was carried out with model drugs as the alcohol extracted liquid for Radix et Rhizoma Rhei (AELRR) by HPLC under the chromatographic conditions that the column was C18, 4.6 mm x 250 mm, 5 microm; the detection of wavelengths was set at 254 nm; a solution of acetonitrile: 1% acetic acid water (from 0:100 to 100:0) was carried with gradient elution as the mobile phase; the ratio of flow was 1 mL min(-1); the temperature was 40 degrees C. The coefficients were dealt with Excel document. It has been established the expressions that consists of four main parameters: 1) total quantum zero moment as AUC(T), area under curve; 2) total quantum respondent ratio as AUCPW(T), area under curve per weight; 3) total quantum center moment as MCRT(T), mean chromatographic retention time of total quantum, expressed by lambda(T); 4) total quantum variance as VCRT(T), variance of mean chromatographic retention time of total quantum, expressed by sigma2(T), by which we have obtained. The TQSM's parameters of the AELRR, such as AUC(T) as 3.273 x 10(8) microV s, AUCPW(T) as 2.286 x 10(6) microV s mg (-1), MCRT(T) as 33.50 min, VCRT(T) as 484.4 min2, and total quantum concentration as 143.2 mg mL(-1). The total quantum statistic moment can be characterized the curve of chromatographic fingerprints with expressive parameters above, also used to quantitative analyses by AUC(T), to qualitative analyses by AUCPW(T), MCR(T), and VCRT(T). PMID- 18507350 TI - [Absorption of papaverine, laudanosine and cepharanthine across human intestine by using human Caco-2 cells monolayers model]. AB - Absorption of papaverine (PAP), laudanosine (LAU) and cepharanthine (CEP) as some chemical constituents of traditional Chinese medicines in human intestine were studied. By using Caco-2 (the human colonic adenocarcinoma cell lines) cell monolayers as an intestinal epithelial cell model, the permeability of PAP, LAU and CEP were studied from apical side (AP side) to basolateral side (BL side) or from BL side to AP side. The three alkaloids were measured by reversed-phase high performance liquid chromatography coupled with UV detector. Transport parameters and apparent permeability coefficients (Papp) were then calculated and compared with those of propranolol as a control substance of high permeability and atenolol as a control substance of poor permeability. The Papp values of PAP, LAU and CEP were (3.524+/-0.223) x 10(-5), (2.821+/-0.050) x 10(-5) and (6.524+/ 0.052) x 10(-5) cm s(-1) from AP side to BL side, and (5.095+/-0.508) x 10(-5), (2.646+/-0.146) x 10(-5) and (5.495+/-0.036) x 10(-5) cm s(-1) from BL side to AP side, respectively. Their Papp values were identical with those of propranolol, which is a transcellular transport marker. On the other hand, the efflux transport of PAP was 1.45 times higher than its influx transport with 0.69 rate of P(app A-->B)/P(app B-->A). But P(app A-->B)/P(app B-->A) values of LAU and CEP were 1.07 and 1.19, respectively, which suggested that the efflux transport have not been involved in their absorbed mechanism in Caco-2 cells monolayers. There is a good correlation between the Papp value and apparent distribution coefficient (Log D) at pH 7.35 for the three alkaloids. PAP, LAU and CEP can be absorbed across intestinal epithelial cells, and they are completely absorbed compounds. PAP may have been involved in efflux mechanism in Caco-2 cells monolayers model from the basolateral-to-apical direction. The O/W (oil/water) partition coefficient plays key role in their transmembrane permeation. PMID- 18507351 TI - [Characteristics of poloxamer thermosensitive in situ gel of dexamethasone sodium phosphate]. AB - Thermosensitive in situ gel is a novel drug delivery system which can form gel in situ after injection of the polymer solution into the body and releases the drug in a controlled manner, thus provides a promising strategy for localized drug delivery. The aim of the present work is to investigate the characteristics including gelation temperature, sol-gel transition temperature (T(s-g)), gel strength, stable viscosity, erosion and drug release behavior of the thermosensitive in situ gel which are composed of different concentrations of poloxamer Pluronic F127 and F68. The gelation temperature was determined by tube reverse method. Rheological measurements were carried out to evaluate T(s-g), stable viscosity and gel strength. Erosion of the gels and release of dexamethasone sodium phosphate (DSP) from the gels were investigated by membrane free method and HPLC. Increased F127 concentration in gel decreased the gelation temperature, T(s-g) as well as erosion of the gel and drug release rate, while viscosity and gel strength rose accordingly. However, increased F68 in gel could lead to the opposite result. The poloxamer solution below T(s-g) is Newtonian fluid with comparatively low viscosity, but shows the characteristics of the pseudoplastic fluid when temperature rises near to T(s-g). Drug release was controlled by the erosion of the gel matrix, and both of them followed the zero order kinetics. An optimized formation containing 22.5% F127 and 2.5% F68 showed more desirable characteristics which meet the clinical requirements and is of potential in future clinical therapy. PMID- 18507352 TI - [Preliminary study on sea cucumber saponin-nobiliside A liposome and its hemolytic activity]. AB - Nobiliside A (Nob A) liposomes were prepared. Its assay method of content and encapsulation efficiency (EE) were established, and hemolytic activity with Nob A solution in vivo and in vitro were compared. Preparative method, phospholipid content, ratio of phospholipid to cholesterol and ratio of drug to lipids were optimized by single factor exploration. According to the optimized results, 3 batches of Nob A liposomes were prepared, then high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) method was used to determine the content of Nob A and minicolumn centrifugation method to determine EE, transmission electron microscope was used to detect the morphology and laser scatter analysis to evaluate particle sizes of the liposomes. The hemolytic activity was studied both in vivo and in vitro. The results indicated that HPLC-ELSD method and minicolumn centrifugation method used in this study are simple, applicable and accurate for the determination of the content and EE of Nob A liposome respectively . Nob A liposomes have a high EE with spherical shape and uniform size by using the film ultrasonication technique. When the ratio of phospholipid to cholesterol was 2:1 and the ratio of Nob A to lipids was 1:40, the mean EE of Nob A liposomes was 95.7% and the mean diameter was 87.6 nm. Liposomes inhibited the hemolytic activity of Nob A in vivo and in vitro sharply. As for its low hemolytic activity in vivo and in vitro, Nob A liposomes are optimistic to be used by intravenous injection. PMID- 18507353 TI - [Analysis of aristolochic acid A from the aerial and underground parts of Asarum by UPLC-UV]. AB - To develop an UPLC-UV method for the determination of aristolochic acid A in the aerial and underground parts of Asarum, an AcquityUPLC HSST3 column (50 mm x 2.1 mm ID, 1.8 microm) was used with the mobile phase of acetonitrile-0.1% FA and eluted in gradient mode. The flow rate was 0.5 mL min(-1), the detection wavelength was 251 nm, and the column temperature was 40 degrees C. The calibration curve was linear in the range of 18 - 1450 ng of aristolochic acid A with correlation coefficient of 0. 9999 and the minimum detective concentration was 18.25 ng mL (-1). The method developed is accurate and reproducible, and can be applied in the determination of aristolochic acid A in Asarum. PMID- 18507354 TI - [Pharmacokinetic parameter and residua of 63Ni-NiCl2 in rat]. AB - Absorption distribution and excretion of 63Ni-NiCl2, administered orally to rats were studied by using liquid scintillation counting method. It was observed that the concentration-time curves in blood fitted the two compartment model of pharmacokinetics, Ka=6.18 h(-1), T(1/2)alpha =0.79 h, T(1/2)beta=40.68 h, CL =0.42 mL kg(-1) h(-1), Tmax =0.53 h, Cmax=24,987.75 min(-1) mL(-1), and Vd=0.016 L kg(-1). After rats were treated by 63Ni-NiCl2 for 15 days, in 22 tissues tested, the contents of 63Ni-NiCl2 in hair, hypothalamus, hypophysis, pancreas, small and large intestines were higher, and the residua of 63Ni-NiCl2 was not discovered in liver, kidney and heart. Radioactivity eliminated was 83.27% by urine and feces, 54.86% by urine, 28.41% by feces. PMID- 18507355 TI - Mentally ill prisoners in the California Department of Corrections and Rehabilitation: strategies for improving treatment and reducing recidivism. PMID- 18507356 TI - Navigating the Safe Harbor: guidance from the courts on qualifying for the 35 U.S.C. 271(e)(1) exemption from patent infringement of health care related inventions. PMID- 18507357 TI - The Mentally Ill Offender Treatment and Crime Reduction Act and its inappropriate non-violent offender limitation. PMID- 18507358 TI - Unintended consequences: resolving the dilemma of the inadvertent creation of Multiple Employer Welfare Arrangements during business transfers. PMID- 18507359 TI - Staying within the lines: the question of post-stabilization treatment for illegal immigrants under Emergency Medicaid. PMID- 18507360 TI - Organ trafficking and the TVPA: why one word makes a difference in international enforcement efforts. PMID- 18507361 TI - Modeling the kinetics of Fe(II) oxidation in the presence of citrate and salicylate in aqueous solutions at pH 6.0-8.0 and 25 degrees C. AB - The kinetics of Fe(II) oxidation in the presence of low concentrations of citrate and salicylate have been investigated in aqueous solutions over the pH range 6.0 8.0 using colorimetry. A kinetic model has been developed to describe the oxidation of Fe(II) with specific attention given to the oxidation of inorganic Fe(II), formation and dissociation of Fe(II) complexes and the oxidation of these complexes. At low concentrations of salicylate, both experimental data and model show that the common approach to modeling Fe(II) oxidation that assumes pre equilibrium between metal and ligand prior to their oxidation is not valid. Complexation of Fe(II) by salicylate is found to be relatively slow, and oxidation of the complex formed occurs rapidly. Citrate, on the other hand was found to be in rapid equilibrium with Fe(II) but the complex formed was oxidized slowly. Both citrate and salicylate complexes are found to dissociate at a rate much faster than previously thought. A model of the oxidation kinetics of Fe(II) species that incorporates the formation and dissociation kinetics of Fe(II) and Fe(III) complexes of citrate and salicylate as well as the reactions of these species with oxygen and reduced oxygen species including superoxide and hydrogen peroxide provides an excellent description of data obtained over a wide range of concentration and pH conditions. PMID- 18507362 TI - Effect of diffusion on the photoinduced reaction between a tetra-anionic porphyrin and methylviologen cation in methanol. AB - The quenching of the fluorescence decay of electronically excited 5,10,15,20 tetraphenyl- 21H, 23H-porphinetetrasulfonate (TPPS (4-)*) in the presence of methylviologen cations (MV (2+)) was measured at various ionic strengths in methanol. Analysis of the fluorescence decay curves revealed strong evidence for the presence of a second fluorescent species over the entire range of ionic strength used in this work, which is attributed to solvent-separated ion pairs (TPPS (4-)-S-MV (2+)). Transient effects of the fluorescence decays were analyzed, and values for the effective reaction distance, R NH, and the diffusion coefficients, D, were obtained. Diffusion coefficients were independently measured for TPPS (4-) and MV (2+) using the Taylor dispersion method. The values for D obtained by the analysis of the transient effect were found to be smaller than those for the sum of the diffusion coefficients of TPPS (4-) and MV (2+) obtained by the Taylor dispersion method and a possible explanation for this result is given. PMID- 18507363 TI - Probing the photoexcited states of rhodium corroles by time-resolved Q-band EPR. Observation of strong spin-orbit coupling effects. AB - The photoexcited states of two 5,10,15-tris(pentafluorophenyl)corroles (tpfc), hosting Rh(III) in their core, namely Rh(pyr)(PPh 3)(tpfc) and Rh(PPh 3)(tpfc), have been studied by time-resolved electron paramagnetic resonance (TREPR) combined with pulsed laser excitation. Using the transient nutation technique, the spin polarized spectra are assigned to photoexcited triplet states. The spectral widths observed for the two Rh(III) corroles crucially depend on the axial ligands at the Rh(III) metal ion. In case of Rh(PPh 3)(tpfc), the TREPR spectra are found to extend over 200 mT, which exceeds the spectral width of non transition-metal corroles by more than a factor of 3. Moreover, the EPR lines of the Rh(III) corroles are less symmetric than those of the non-transition-metal corrroles. The peculiarities in the TREPR spectra of the Rh(III) corroles can be rationalized in terms of strong spin-orbit coupling (SOC) associated with the transition-metal character of the Rh(III) ion. It is assumed that SOC in the photoexcited Rh(III) corroles effectively admixes metal centered (3)dd-states to the corrole centered (3)pipi*-states detected in the TREPR experiments. This admixture leads to an increased zero-field splitting and a large g-tensor anisotropy as manifested by the excited Rh(III) corroles. PMID- 18507364 TI - Molecular dynamics simulations of perylene and tetracene librations: comparison with femtosecond upconversion data. AB - In a prior manuscript by Xu et al. [Xu, J.; Shen, X.; Knutson, J. R. J. Phys. Chem. A 2003, 107, 8383], time-resolved fluorescence emission anisotropy measurements were performed on perylene and tetracene in hexadecane using an upconversion technique with approximately 100 fs resolution. The anisotropy transients contained previously unseen decay terms of approximately 300 fs. In perylene, their amplitude corresponded to the " r(o) defect" that has gathered interest over decades. We ascribed this term to a predominantly in-plane libration. In this manuscript, we present molecular dynamics simulations for the motions of perylene and tetracene using the CHARMm molecular dynamics program (version c29b2). Both rotational correlation functions contain subpicosecond decay terms that resemble experimental anisotropy decays. It was suggested that the r(o) defect might arise from excited-state distortions of perylene, so we conducted quantum mechanical calculations to show that such distortion does not significantly displace the oscillators. We compare the case of perylene, with a strongly allowed singlet emission transition, to that of the weakly allowed tetracene transition. In perylene, motion alone can explain subpicosecond anisotropy decay, while tetracene decay also contains vibrational coupling terms, as previously reported by Sarkar et al. [Sarkar, N.; Takeuchi, S.; Tahara, T. J. Phys. Chem. A 1999, 103, 4808]. PMID- 18507365 TI - Emergence of charge-transfer-to-solvent band in the absorption spectra of hydrogen halides on ice nanoparticles: spectroscopic evidence for acidic dissociation. AB - Extensive ab initio calculations complemented by a photodissociation experiment at 193 nm elucidate the nature of hydrogen halide molecules bound on free ice nanoparticles. Electronic absorption spectra of small water clusters (up to 5 water molecules) and water clusters doped with hydrogen fluoride, hydrogen chloride and hydrogen bromide were calculated. The spectra were modeled at the time-dependent density functional (TDDFT) level of theory with the BHandHLYP functional using the reflection principle. We observe the emergence of a charge transfer-to-solvent (CTTS) band in the absorption spectra upon the acidic dissociation of the hydrogen halides. The CTTS band provides a spectroscopically observable feature for the acidic dissociation. The calculated spectra were compared with our new experimental photodissociation data for larger water clusters doped with HCl and HBr. We conclude that HCl and HBr dissociate to a large extent on the surface of ice nanoparticles at temperatures near 120 K and photoactive products are formed. The acidic dissociation of HX leads to an enhancement by about 4 orders of magnitude of the HCl photolysis rate in the 200 300 nm region, which is potentially relevant for the halogen budget in the atmosphere. PMID- 18507366 TI - Synthesis and biological evaluation of D-amino acid oxidase inhibitors. AB - D-amino acid oxidase (DAAO) catalyzes the oxidation of D-amino acids including d serine, a full agonist at the glycine site of the NMDA receptor. A series of benzo[ d]isoxazol-3-ol derivatives were synthesized and evaluated as DAAO inhibitors. Among them, 5-chloro-benzo[ d]isoxazol-3-ol (CBIO) potently inhibited DAAO with an IC50 in the submicromolar range. Oral administration of CBIO in conjunction with d-serine enhanced the plasma and brain levels of d-serine in rats compared to the oral administration of d-serine alone. PMID- 18507367 TI - Flavonoids for controlling starch digestion: structural requirements for inhibiting human alpha-amylase. AB - In this study we investigated the structural requirements for inhibition of human salivary alpha-amylase by flavonoids. Four flavonols and three flavones, out of the 19 flavonoids tested, exhibited IC50 values less than 100 microM against human salivary alpha-amylase activity. Structure-activity relationships of these inhibitors by computational ligand docking showed that the inhibitory activity of flavonols and flavones depends on (i) hydrogen bonds between the hydroxyl groups of the polyphenol ligands and the catalytic residues of the binding site and (ii) formation of a conjugated pi-system that stabilizes the interaction with the active site. Our findings show that certain naturally occurring flavonoids act as inhibitors of human alpha-amylase, which makes them promising candidates for controlling the digestion of starch and postprandial glycemia. PMID- 18507368 TI - Synthesis, cytotoxicity, DNA interaction, and topoisomerase II inhibition properties of novel indeno[2,1-c]quinolin-7-one and indeno[1,2-c]isoquinolin-5,11 dione derivatives. AB - Indeno[2,1- c]quinolin-7-ones and 6 H-indeno[1,2- c]isoquinolin-5,11-diones, bearing two cationic aminoalkyl side chains, were synthesized and evaluated for DNA interaction, topoisomerases inhibition, and cytotoxicity against human cancer cell lines. They displayed strong interaction with DNA and one indeno[1,2- c]isoquinolin-5,11-dione bearing side chains at N-6 and C-8 positions ( 6a) was a potent human topoisomerase II inhibitor with high cytotoxicity toward HL60 cells. An increased topoisomerase II inhibition is found with (a) a cationic aminoalkyl side chain at the C-8 rather than at the C-9 position, (b) a dimethylaminoethoxy side chain at the C-8 position introduced on the N-6 monosubstituted derivative, going with suppression of topoisomerase I poisoning, and (c) a dimethylaminoethyl rather than a dimethylaminopropyl side chain at the N-6 position. The cytotoxicity was only partially reduced when using the topoisomerase II-mutated mitoxantrone-resistant HL60/MX2 cell line, suggesting that additional targets are involved in their mechanism of action. These indeno[1,2- c]isoquinolin-5,11-dione derivatives represent new DNA-topoisomerase II interfering anticancer molecules. PMID- 18507369 TI - Designing selective, high affinity ligands of 5-HT1D receptor by covalent dimerization of 5-HT1F ligands derived from 4-fluoro-N-[3-(1-methyl-4 piperidinyl)-1H-indol-5-yl]benzamide. AB - We demonstrate here that covalent dimerization of 5-HT 1 ligands is an effective design strategy to modulate affinity and selectivity of 5-HT 1 ligands. This approach was applied to LY-334370, a selective agonist of 5-HT 1F receptor, to generate structurally well-defined divalent molecules. Radioligand binding assays to three cloned 5-HT 1 receptor subtypes (5-HT 1B, 5-HT 1D, 5-HT 1F) demonstrated that the affinity of a series of homologous dimers varied significantly upon exploration of three structural variables (linker length, attachment position, functionality). In particular, the series of C 3-to-C 3 linked dimers derived from a monomer ( 3) showed high binding affinity to 5-HT 1D (for example, K i approximately 0.3 nM for dimer 8) but did not bind to 5-HT 1F ( K i > 0.01 mM), providing >10000-fold subtype selectivity. Results from a functional assay (rabbit saphenous vein contraction) demonstrate that certain dimers are 5-HT 1 receptor agonists. PMID- 18507370 TI - Homology modeling and site-directed mutagenesis to identify selective inhibitors of endothelin-converting enzyme-2. AB - Endothelin-converting enzyme-2 (ECE-2), a member of M13 family of zinc metallopeptidases, has previously been shown to process a number of neuropeptides including those derived from prodynorphin, proenkephalin, proSAAS, and amyloid precursor protein. ECE-2, unlike ECE-1, exhibits restricted neuroendocrine distribution and acidic pH optimum; it is consistent with a role in the regulation of neuropeptide levels in vivo. Here, we report the generation of a three-dimensional (3D) molecular model of ECE-2 using the crystal structure of neprilysin (EC 3.4.24.11) as a template. On the basis of the predictions made from the molecular model, we mutated and tested two residues, Trp 148 and Tyr 563, in the catalytic site. The mutation of Tyr 563 was found to significantly affect the catalytic activity and inhibitor binding. The molecular model was used to virtually screen a small molecule library of 13 000 compounds. Among the top scoring compounds three were found to inhibit ECE-2 with high affinity and exhibited specificity for ECE-2 compared to neprilysin. Thus, the model provides a new useful tool to probe the active site of ECE-2 and design additional selective inhibitors of this enzyme. PMID- 18507371 TI - Discovery of imidazo[1,5-c]imidazol-3-ones: weakly basic, orally active factor Xa inhibitors. AB - The coagulation enzyme factor Xa (FXa) has been recognized as a promising target for the development of new antithrombotic agents. We previously found compound 1 to be an orally bioavailable FXa inhibitor in fasted monkeys; however, 1 showed poor bioavailability in rats and fed monkeys. To work out the pharmacokinetic problems, we focused our synthetic efforts on the chemical conversion of the 4 (imidazo[1,2- a]pyridin-5-yl)piperazine moiety of 1 to imidazolylpiperidine derivatives (fused and nonfused), which resulted in the discovery of the weakly basic imidazo[1,5- c]imidazol-3-one 3q as a potent and selective FXa inhibitor. Compound 3q showed favorable oral bioavailability in rats and monkeys under both fasted and fed conditions and antithrombotic efficacy in a rat model of venous thrombosis after oral administration, without a significant increase in bleeding time (unlike warfarin). On the basis of these promising properties, compound 3q was selected for further evaluation. PMID- 18507373 TI - Membrane-interaction quantitative structure--activity relationship (MI-QSAR) analyses of skin penetration enhancers. AB - Membrane-interaction quantitative structure-activity relationship (MI-QSAR) models for two skin penetration enhancer data sets of 61 and 42 compounds were constructed and compared to QSAR models constructed for the same two data sets using only classic intramolecular QSAR descriptors. These two data sets involve skin penetration enhancement of hydrocortisone and hydrocortisone acetate, and the enhancers are generally similar in structure to lipids and surfactants. A new MI-QSAR descriptor, the difference in the integrated cylindrical distribution functions over the phospholipid monolayer model, in and out of the presence of the skin penetration enhancer, DeltaSigma h(r), was developed. This descriptor is dominant in the optimized MI-QSAR models of both training sets studied and greatly reduces the size and complexity of the MI-QSAR models as compared to those QSAR models developed using the classic intramolecular descriptors. The MI QSAR models indicate that good penetration enhancers make bigger "holes" in the monolayer and are less aqueous-soluble, so as to preferentially enter the monolayer, than are poor penetration enhancers. The skin penetration enhancer thus alters the structure and organization of the monolayer. This space and time alteration in the structure and dynamics of the membrane monolayer is captured by DeltaSigma h(r) and is simplistically referred to as "holes" in the monolayer. The MI-QSAR models explain 70-80% of the variance in skin penetration enhancement across each of the two training sets and are stable predictive models using accepted diagnostic measures of robustness and predictivity. PMID- 18507372 TI - Synthesis and quantitative structure-activity relationship of fatty acid amide hydrolase inhibitors: modulation at the N-portion of biphenyl-3-yl alkylcarbamates. AB - Alkylcarbamic acid biphenyl-3-yl esters are a class of fatty acid amide hydrolase (FAAH) inhibitors that comprises cyclohexylcarbamic acid 3'-carbamoylbiphenyl-3 yl ester (URB597), a compound with analgesic, anxiolytic-like and antidepressant like properties in rat and mouse models. Here, we extended the structure-activity relationships (SARs) for this class of compounds by replacing the cyclohexyl ring of the parent compound cyclohexylcarbamic acid biphenyl-3-yl ester (URB524) (FAAH IC50 = 63 nM) with a selected set of substituents of different size, shape, flexibility, and lipophilicity. Docking experiments and linear interaction energy (LIE) calculations indicated that the N-terminal group of O-arylcarbamates fits within the lipophilic region of the substrate-binding site, mimicking the arachidonoyl chain of anandamide. Significant potency improvements were observed for the beta-naphthylmethyl derivative 4q (IC50 = 5.3 nM) and its 3' carbamoylbiphenyl-3-yl ester 4z (URB880, IC50 = 0.63 nM), indicating that shape complementarity and hydrogen bonds are crucial to obtain highly potent inhibitors. PMID- 18507374 TI - Novel chelate ring-opening induced by silver(I) of five-coordinate palladium(II) and platinum(II) complexes containing tripodal polyphosphines. AB - The ionic complexes [Pd(NP 3)X]X [NP 3 = tris[2-(diphenylphosphino)ethyl]amine, X = Cl (1), Br(2)] and [M(PP 3)X]X [PP 3 = tris[2 (diphenylphosphino)ethyl]phosphine, M = Pd, X = Cl (3), Br(4); M = Pt, X = Cl (5), Br (6)] contain square pyramidal (1, 2) and trigonal bipyramidal (3- 6) cations with three fused chelate rings to M and one M-X bond. By addition of AgX salts (X = Cl, Br, NO 3) an unexpected ring-opening reaction occurs with formation of the heteronuclear species PdAg(NP 3)X 3 [X = Cl (7), Br (8)], MAg(PP 3)X 3 [M = Pd, X = Cl (9), Br (10), NO 3 (13);M = Pt, X = Cl (11), Br (12), NO 3 (14)]. The complexes have been characterized in the solid state and solution. The X-ray crystal structures of 9 and 13 reveal a distorted square-planar arrangement to Pd(II) that is coordinated to three P of PP 3 (the central and two terminal atoms) and to one chloride (9) or one oxygen atom of NO 3 (13). The resultant dangling phosphorus of the ring opening is bound to Ag(I) that completes the three- [PAgCl 2 ( 9)] and four-coordination [PAg(ONO 2)(O 2NO) (13)] through the donor atoms of the anions with the nitrates in 13 unusually acting as both mono- and bidentate ligands. Complexes 7, 8, 10, and 11 undergo oligomerization in solution. Complex 10 oligomerizes giving rise to the ionic compound [Pd 4Ag 2(PP 3) 2 Br 9]Br ( 10a) whose X-ray crystal structure indicates the presence of cations with a Pd(mu-Br) 3Pd unit that connects via bromide bridges two BrPdP 2PPAg Br 2 fragments containing distorted square-planar and trigonal-planar Pd(II) and Ag(I) centers, respectively. The palladium(II) metal centers in the central unit afford the five-coordination (PdBr 5) with a distorted trigonal bipyramidal geometry. The ionic system [Pt 2Ag 2(PP 3) 2 Cl 5]Cl (11a) consists of chloride anions and heteronuclear monocations. The X-ray crystal structure reveals that the cations contain two distorted square-planar ClPtP 3 units bridged by one PAgCl(mu-Cl) 2AgP fragment that is bearing tetrahedral (PAgCl 3) and trigonal planar PAgCl 2 silver(I) centers. Further additions of the corresponding AgX salts to complexes 7- 14 did not give rise to any new ring opening reaction. PMID- 18507375 TI - Metal fluorides form strong hydrogen bonds and halogen bonds: measuring interaction enthalpies and entropies in solution. AB - The organometallic compound trans-(tetrafluoropyrid-2-yl)bis(triethylphosphine) fluoronickel(II) (NiF) is shown to serve as a strong hydrogen bond and halogen bond acceptor in solution via intermolecular interactions with the fluoride ligand. The nature of the interactions has been confirmed by multinuclear NMR spectroscopy. Experimental binding constants, enthalpies, and entropies of interaction with hydrogen-bond-donor indole and halogen-bond-donor iodopentafluorobenzene have been determined by 19F NMR titration. In toluene-d8 solution indole forms a 1:1 and 2:1 complex with NiF (K1 = 57.9(3), K2 = 0.58(4)). Interaction enthalpies and entropies are -23.4(2) kJ mol-1 and -44.5(8) J mol-1 K-1, respectively, for the 1:1 complex; -14.8(8) kJ mol-1 and -53(3) J mol-1 K-1, respectively, for the 2:1 complex. In toluene-d8 solution iodopentafluorobenzene forms only a 1:1 complex (K1 = 3.41(9)) with enthalpy and entropy of interaction of -16(1) kJ mol-1 and -42(4) J mol-1 K-1, respectively. A marked solvent effect was observed for the halogen bond interaction. NMR titrations in heptane solution indicated formation of both 1:1 and 2:1 complexes of iodopentafluorobenzene with NiF (K1 = 21.8(2), K2 = 0.22(4)). Interaction enthalpies and entropies are -26(1) kJ mol-1 and -63(4) J mol-1 K-1, respectively, for the 1:1 complex; -21(1) kJ mol-1 and -83(5) J mol-1 K-1, respectively, for the 2:1 complex. There is a paucity of such experimental energetic data particularly for halogen bonds despite substantial structural data. These measurements demonstrate that halogen bonds are competitive with hydrogen bonds as intermolecular interactions and provide a suitable benchmark for theoretical calculations and quantitative input into design efforts in supramolecular chemistry and crystal engineering. PMID- 18507376 TI - Quantitative conformational analysis of partially folded proteins from residual dipolar couplings: application to the molecular recognition element of Sendai virus nucleoprotein. AB - A significant fraction of proteins coded in the human proteome do not fold into stable three-dimensional structures but are either partially or completely unfolded. A key feature of this family of proteins is their proposed capacity to undergo a disorder-to-order transition upon interaction with a physiological partner. The mechanisms governing protein folding upon interaction, in particular the extent to which recognition elements are preconfigured prior to formation of molecular complexes, can prove difficult to resolve in highly flexible systems. Here, we develop a conformational model of this type of protein, using an explicit description of the unfolded state, specifically modified to allow for the presence of transient secondary structure, and combining this with extensive measurement of residual dipolar couplings throughout the chain. This combination of techniques allows us to quantitatively analyze the level and nature of helical sampling present in the interaction site of the partially folded C-terminal domain of Sendai virus nucleoprotein (N(TAIL)). Rather than fraying randomly, the molecular recognition element of N(TAIL) preferentially populates three specific overlapping helical conformers, each stabilized by an N-capping interaction. The unfolded strands adjacent to the helix are thereby projected in the direction of the partner protein, identifying a mechanism by which they could achieve nonspecific encounter interactions prior to binding. This study provides experimental evidence for the molecular basis of helix formation in partially folded peptide chains, carrying clear implications for understanding early steps of protein folding. PMID- 18507378 TI - Synthesis of all-conjugated diblock copolymers by quasi-living polymerization and observation of their microphase separation. AB - We designed and synthesized the all-conjugated diblock copolymers poly(3 hexylthiophene-block-3-(2-ethylhexyl)thiophene)s (P(3HT-b-3EHT)s) via a modified Grignard metathesis (GRIM), a type of quasi-living polymerization, and studied their microphase-separated structures. The P(3HT-b-3EHT)s synthesized had well controlled molecular weights and very narrow polydispersity indices (PDIs), which demonstrates the usefulness of GRIM polymerization for the synthesis of semiconducting block copolymers. P(3HT-b-3EHT)s self-organized to form clear microphase-separated patterns upon thermal treatment, as observed by AFM. Interestingly, the enhancement of the interchain interaction of the P3HT segments compared with the P3HT homopolymer was clearly observed from the UV-vis spectra, despite the fact that the amount of crystalline P3HT fraction was reduced to 83% of the total polymer amount in P(3HT-b-3EHT). It is suggested that the relatively unconstrained, amorphous segments of P3EHT can enhance the crystallization of P3HT segments to form an ordered self-organized nanostructure. PMID- 18507379 TI - Total synthesis and molecular target of largazole, a histone deacetylase inhibitor. AB - Full details of the concise and convergent synthesis (eight steps, 19% overall yield), its extension to the preparation of a series of key analogues, and the molecular target and pharmacophore of largazole are described. Central to the synthesis of largazole is a macrocyclization reaction for formation of the strained 16-membered depsipeptide core followed by an olefin cross-metathesis reaction for installation of the thioester. The biological evaluation of largazole and its key analogues, including an acetyl analogue, a thiol analogue, and a hydroxyl analogue, suggested that histone deacetylases (HDACs) are molecular targets of largazole and largazole is a class I HDAC inhibitor. In addition, structure-activity relationship (SAR) studies revealed that the thiol group is the pharmacophore of the natural product. Largazole's HDAC inhibitory activity correlates with its antiproliferative activity. PMID- 18507380 TI - Trapping and structural elucidation of a very advanced intermediate in the lesion extrusion pathway of hOGG1. AB - Here we present the first structure of a very advanced intermediate in the lesion extrusion pathway of a DNA glycosylase, human 8-oxoguanine DNA glycosylase (hOGG1), and a substrate DNA containing a mutagenic lesion, 8-oxoguanine (oxoG). The structure was obtained by irradiation and flash-freezing of a disulfide-cross linked (DXLed) complex of hOgg1 bound to DNA containing a novel photocaged derivative of oxoG. The X-ray structure reveals that, upon irradiation, the oxoG lesion has transited from the exosite to the active site pocket, but has not undergone cleavage by the enzyme. Furthermore, all but one of the specificity determining interactions between the lesion and the enzyme are unformed in the flashed complex (FC), because active site functionality and elements of the DNA backbone are mispositioned. This structure thus provides a first glimpse into the structure of a very late-stage intermediate in the lesion-extrusion pathway--the latest observed to date for any glycosylase--in which the oxoG has undergone insertion into the enzyme active site following photodeprotection, but the enzyme and DNA have not yet completed the slower process of adjusting to the presence of the lesion in the active site. PMID- 18507381 TI - Evidence of an intermediate and parallel pathways in protein unfolding from single-molecule fluorescence. AB - Determining how proteins fold into their native structures is a subject of great importance, since ultimately it will allow protein structure and function to be predicted from primary sequence data. In addition, there is now a clear link between protein unfolding and misfolding events and many disease states. However, since proteins fold over rugged, multidimensional energy landscapes, this is a challenging experimental and theoretical problem. Single-molecule fluorescence methods developed over the past decade have the potential to follow the unfolding/folding of individual molecules. Mapping out the landscape without ensemble averaging will enable the identification of intermediate states which may not be significantly populated, in addition to the presence of multiple pathways. To date, there have been only a limited number of single-molecule folding/unfolding studies under nonequilibrium conditions and no intermediates have been observed. Here, for the first time, we present a single-molecule study of the unfolding of a large autofluorescent protein, Citrine, a variant of green fluorescent protein. Single-molecule fluorescence techniques are used to directly detect an intermediate on the unfolding/folding pathway and the existence of parallel unfolding pathways. This work, and the novel methods used, shows that single-molecule fluorescence can now provide new, hitherto experimentally inaccessible, insights into the folding/unfolding of proteins. PMID- 18507383 TI - Artificial metalloenzyme for enantioselective sulfoxidation based on vanadyl loaded streptavidin. AB - Nature's catalysts are specifically evolved to carry out efficient and selective reactions. Recent developments in biotechnology have allowed the rapid optimization of existing enzymes for enantioselective processes. However, the ex nihilo creation of catalytic activity from a noncatalytic protein scaffold remains very challenging. Herein, we describe the creation of an artificial enzyme upon incorporation of a vanadyl ion into the biotin-binding pocket of streptavidin, a protein devoid of catalytic activity. The resulting artificial metalloenzyme catalyzes the enantioselective oxidation of prochiral sulfides with good enantioselectivities both for dialkyl and alkyl-aryl substrates (up to 93% enantiomeric excess). Electron paragmagnetic resonance spectroscopy, chemical modification, and mutagenesis studies suggest that the vanadyl ion is located within the biotin-binding pocket and interacts only via second coordination sphere contacts with streptavidin. PMID- 18507382 TI - Enantiomer-specific binding of ruthenium(II) molecular wires by the amine oxidase of Arthrobacter globiformis. AB - The copper amine oxidase from Arthrobacter globiformis (AGAO) is reversibly inhibited by molecular wires comprising a Ru(II) complex head group and an aromatic tail group joined by an alkane linker. The crystal structures of a series of Ru(II)-wire-AGAO complexes differing with respect to the length of the alkane linker have been determined. All wires lie in the AGAO active-site channel, with their aromatic tail group in contact with the trihydroxyphenylalanine quinone (TPQ) cofactor of the enzyme. The TPQ cofactor is consistently in its active ("off-Cu") conformation, and the side chain of the so called "gate" residue Tyr296 is consistently in the "gate-open" conformation. Among the wires tested, the most stable complex is produced when the wire has a (CH2)4- linker. In this complex, the Ru(II)(phen)(bpy)2 head group is level with the protein molecular surface. Crystal structures of AGAO in complex with optically pure forms of the C4 wire show that the linker and head group in the two enantiomers occupy slightly different positions in the active-site channel. Both the Lambda and Delta isomers are effective competitive inhibitors of amine oxidation. Remarkably, inhibition by the C4 wire shows a high degree of selectivity for AGAO in comparison with other copper-containing amine oxidases. PMID- 18507384 TI - Origin of selectivity switch in Fischer-Tropsch synthesis over Ru and Rh from first-principles statistical mechanics studies. AB - For its unique position in chemical industry, Fischer-Tropsch (FT) synthesis has been a hot subject in heterogeneous catalysis. Due to its great complexity in product distribution, it remains unclear how to maximally convert syngas to long chain hydrocarbons. By combining extensive DFT calculations with grand canonical Monte Carlo simulations, this work examines the key elementary steps in FT synthesis over Ru and Rh surfaces, including CO dissociation, C/C coupling, and hydrogenations. The origin of the relationship between activity and selectivity of catalysts is revealed based on the calculated reaction rate at working temperatures, in which the catalytic role of surface steps as the center of accumulating surface CH(x) species is highlighted. This theoretical work demonstrates that the ability to dissociate CO under carbon-rich conditions is the key requirement for a good FT catalyst. The RC + C (R = alkyl or H) pathway occurring at surface steps may be a general mechanism for FT chain propagation on transition metals. PMID- 18507385 TI - Effective Pd-nanoparticle (PdNP)-catalyzed Negishi coupling involving alkylzinc reagents at room temperature. AB - Pd(OAc)(2) is an efficient catalyst precursor for Negishi coupling in the presence of Bu(4)NBr. Secondary and primary alkylzinc reagents with beta-H and arylzinc reagents all reacted with aryl iodides at temperatures as low as -20 degrees C, giving moderate to good yields. One example of coupling between alkynylzinc reagents and aryl iodides was tested and the yield was good. Preliminary kinetic studies indicated that the process involved PdNPs as the active catalytic species. PMID- 18507386 TI - N-propargylic beta-enaminones: common intermediates for the synthesis of polysubstituted pyrroles and pyridines. AB - N-Propargylic beta-enaminones have been used as common intermediates for the synthesis of polysubstituted pyrroles and pyridines. Best results have been obtained using DMSO as solvent. In the presence of Cs(2)CO(3) N-propargylic beta enaminones are cyclized to pyrroles in good to high yields, whereas omitting bases and using CuBr leads to the selective formation of pyridines. PMID- 18507387 TI - Iron-catalyzed intramolecular o-arylation: synthesis of 2-aryl benzoxazoles. AB - A practical iron-catalyzed intramolecular O-arylation reaction and its application in the synthesis of benzoxazole derivatives, starting from the readily available 2-haloanilines, is presented. The key cyclization step involves the use of a combination of the cheap and environmentally friendly FeCl(3) and 2,2,6,6-tetramethyl-3,5 heptanedione (TMHD) as the catalyst system. PMID- 18507388 TI - Palladium-catalyzed chloroimination of imidoyl chlorides to a triple bond: an intramolecular reaction leading to 4-chloroquinolines. AB - In this paper, a new type of effective chloroimination was reported. This reaction afforded 4-chloro-2-perfluoroalkyl quinolines from fluorinated imidoyl chlorides in high yields. This is the first achievement of oxidative addition reductive elimination type C-Cl bond activation by chloropalladation. PMID- 18507389 TI - Oligo(p-phenyleneethynylene)s with hydrogen-bonded coplanar conformation. AB - A series of monodispersed oligo( p-phenyleneethynylene)s were synthesized bearing intramolecular hydrogen bonds between side chains of adjacent phenylene units in the backbone. Thus, all repeating units of the molecules are constrained in a coplanar orientation. Such planarized conformation is considered favorable for single-molecule conductance. Photophysical characterization results show narrowed bandgaps and extended conjugation lengths, consistent with a rigid, planar backbone framework as a result of intramolecular hydrogen bonding. PMID- 18507390 TI - Base-mediated cyclization reaction of 2-alkynylphenylphosphine oxides: synthesis and photophysical properties of benzo[b]phosphole oxides. AB - The base-mediated intramolecular cyclization reaction of 2-alkynylphenylphosphine oxides affords benzo[ b]phosphole oxides, which show intense blue-green fluorescence. Benzo[ b]phospholes are also prepared by the reduction of benzo[ b]phosphole oxides. PMID- 18507391 TI - Copper(I)-catalyzed carboxylation of aryl- and alkenylboronic esters. AB - The copper(I)-catalyzed carboxylation reaction of aryl- and alkenylboronic esters proceeded smoothly under CO(2) to give the corresponding carboxylic acid in good yield. This reaction showed wide generality with higher functional group tolerance compared to the corresponding Rh(I)-catalyzed reaction. PMID- 18507392 TI - Origin of enantioselection in Hetero-Diels-Alder reactions catalyzed by naphthyl TADDOL. AB - The asymmetric hetero-Diels-Alder reaction of benzaldehyde with 1-dimethylamino-3 tert-butyldimethylsiloxy butadiene catalyzed by ( R, R)-1-Np-TADDOL was studied using computational methods. A theoretical rationale was developed through the combined use of molecular mechanics and ONIOM(B3LYP/6-31G(d):AM1) calculations. The origins of stereoselection in this process were identified, and excellent correlation between experiment and theory was found. PMID- 18507393 TI - Assembly of isoquinolines via CuI-catalyzed coupling of beta-keto esters and 2 halobenzylamines. AB - CuI-catalyzed coupling of 2-halobenzylamines with beta-keto esters or 1,3 diketones in i-PrOH under the action of K(2)CO(3) produced 1,2 dihydroisoquinolines as the coupling/condensative cyclization products, which underwent smooth dehydrogenation under air atmosphere to afford substituted isoquinolines. PMID- 18507394 TI - Using toluates as simple and versatile radical precursors. AB - The viability of the toluate moiety as a radical precursor has been examined by studying deoxygenation and cyclization reactions. PMID- 18507395 TI - Engineering substrate preference in subtilisin: structural and kinetic analysis of a specificity mutant. AB - Bacillus subtilisin has been a popular model protein for engineering altered substrate specificity. Although some studies have succeeded in increasing the specificity of subtilisin, they also demonstrate that high specificity is difficult to achieve solely by engineering selective substrate binding. In this paper, we analyze the structure and transient state kinetic behavior of Sbt160, a subtilisin engineered to strongly prefer substrates with phenylalanine or tyrosine at the P4 position. As in previous studies, we measure improvements in substrate affinity and overall specificity. Structural analysis of an inactive version of Sbt160 in complex with its cognate substrate reveals improved interactions at the S4 subsite with a P4 tyrosine. Comparison of transient state kinetic behavior against an optimal sequence (DFKAM) and a similar, but suboptimal, sequence (DVRAF) reveals the kinetic and thermodynamic basis for increased specificity, as well as the limitations of this approach. While highly selective substrate binding is achieved in Sbt160, several factors cause sequence specificity to fall short of that observed with natural processing subtilisins. First, for substrate sequences which are nearly optimal, the acylation reaction becomes faster than substrate dissociation. As a result, the level of discrimination among these substrates diminishes due to the coupling between substrate binding and the first chemical step (acylation). Second, although Sbt160 has 24-fold higher substrate affinity for the optimal substrate DFKAM than for DVRAF, the increased substrate binding energy is not translated into improved transition state stabilization of the acylation reaction. Finally, as interactions at subsites become stronger, the rate-determining step in peptide hydrolysis changes from acylation to product release. Thus, the release of the product becomes sluggish and leads to a low k(cat) for the reaction. This also leads to strong product inhibition of substrate turnover as the reaction progresses. The structural and kinetic analysis reveals that differences in the binding modes at subsites for substrates, transition states, and products are subtle and difficult to manipulate via straightforward protein engineering. These findings suggest several new strategies for engineering highly sequence selective enzymes. PMID- 18507396 TI - Role of the N- and C-terminal domains in binding of apolipoprotein E isoforms to heparan sulfate and dermatan sulfate: a surface plasmon resonance study. AB - The ability of apolipoprotein E (apoE) to bind to cell-surface glycosaminoglycans (GAGs) is important for lipoprotein remnant catabolism. Using surface plasmon resonance, we previously showed that the binding of apoE to heparin is a two-step process; the initial binding involves fast electrostatic interaction, followed by a slower hydrophobic interaction. Here we examined the contributions of the N- and C-terminal domains to each step of the binding of apoE isoforms to heparan sulfate (HS) and dermatan sulfate (DS). ApoE3 bound to less sulfated HS and DS with a decreased favorable free energy of binding in the first step compared to heparin, indicating that the degree of sulfation has a major effect on the electrostatic interaction of GAGs with apoE. Mutation of a key Lys residue in the N-terminal heparin binding site of apoE significantly affected this electrostatic interaction. Progressive truncation of the C-terminal alpha-helical regions which favors the monomeric form of apoE3 greatly weakened the ability of apoE3 to bind to HS, with a much reduced favorable free energy of binding of the first step, suggesting that the C-terminal domain contributes to the GAG binding of apoE by the oligomerization effect. In agreement with this, dimerization of the apoE3 N terminal fragment via disulfide linkage restored the electrostatic interaction of apoE with HS. Significantly, apoE4 exhibited much stronger binding to HS and DS than apoE2 or apoE3 in both lipid-free and lipidated states, perhaps resulting from enhanced electrostatic interaction through the N-terminal domain. This isoform difference in GAG binding of apoE may be physiologically significant such as in the retention of apoE-containing lipoproteins in the arterial wall. PMID- 18507397 TI - Direct Loading of the purified endogenous inhibitor into the cytoplasm of patched cardiomyocytes blocks the ion currents and calcium transport through the NCX1 protein. AB - The Na(+)-Ca(2+) exchanger in mammalian heart muscle (NCX1) is the central transporter protein that regulates extrusion of Ca(2+) from the heart cell. However, the functional biochemistry and physiology of NCX1 have been severely hampered by the absence of any specific high-affinity inhibitor. Here we describe advanced procedures for purifying a candidate inhibitor, previously called endogenous inhibitor factor (NCX(IF)), and demonstrate its direct actions on NCX1 activities in the single-cell system. A combination of advanced HILIC (hydrophilic interaction liquid chromatography) procedures with analytical tests suggests that the properties of NCX(IF) resemble those of a small (disaccharide size) polar molecule lacking any aromatic rings, conjugated bonds, or a primary amino group. The effects of NCX(IF) on the NCX1-mediated ion currents (I(NCX)) and cytosolic Ca(2+) extrusion were detected by a combination of patch-clamp and confocal microscopy under conditions in which the purified NCX(IF) was directly loaded into the cytoplasm of patched cardiomyocytes. It was demonstrated that cytosolic NCX(IF) blocks the Ca(2+)-activated NCX1 inward current and the accompanying extrusion of Ca(2+) from the cell with high efficacy. A constant fraction of NCX1 inhibition was observed under conditions in which the cytosolic [Ca(2+)](i) was varied at fixed doses of NCX(IF), suggesting that the degree of inhibition is controlled by NCX(IF) dose and not by cytosolic Ca(2+) concentration. NCX(IF) blocks equally well both the Ca(2+) extrusion and Ca(2+) entry modes of NCX1, consistent with thermodynamic principles expected for the functioning of a bidirectional "carrier-type" transport system. We concluded that NCX(IF) interacts with a putative regulatory domain from the cytosolic side and, thus, may play an important regulatory role in controlling Ca(2+) signaling in the heart. This may represent a new potential tool for developing novel treatments for cardiac Ca(2+) signaling dysfunction. PMID- 18507398 TI - Impact of the enfuvirtide resistance mutation N43D and the associated baseline polymorphism E137K on peptide sensitivity and six-helix bundle structure. AB - Enfuvirtide (ENF), the first human immunodeficiency virus type 1 (HIV-1) fusion inhibitor approved for clinical use, acts by binding to gp41 heptad repeat 1 (HR1) and preventing its interaction with the viral HR2 region. Treatment emergent resistance to ENF has been mapped to residues within HR1, and these mutations decrease its susceptibility to ENF and may reduce viral fitness and pathogenesis, although the mechanism for these effects is not clear. N43D, a common ENF resistance mutation, was found in in vitro assays to cause a 5-50-fold in antiviral activity. We introduced this mutation into peptide models and determined the impact of this mutation by circular dichroism and X-ray crystallography. We find that the mutation results in a decrease in the thermal stability of the six-helix bundle and causes a significant change in the HR1-HR2 interface, including a loss of HR2 helicity. These data form a mechanistic basis for the decrease in ENF sensitivity and six-helix bundle stability. The E137K polymorphism, generally present at baseline in patients who develop N43D, partially compensates for the loss of stability, and we show that these residues likely form an ion pair. These data form a framework for understanding the impact of resistance mutations on viral fitness and pathogenesis and provide a pathway for the development of novel fusion inhibitor peptides. PMID- 18507399 TI - Multiple surface plasmon resonances and near-infrared field enhancement of gold nanowells. AB - Arrays of Au nanowells (NWs) were fabricated by electron-beam lithography (EBL) and characterized by surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS). It is revealed that these Au NW arrays exhibit multiple SP resonances that can be tuned by adjusting the geometrical characteristics of the NWs. SERS activity of Au NWs was confirmed for a range of excitation wavelengths and a number of model compounds including rhodamine 6G (R6G), phthalazine, and single-stranded oligonucleotides. According to numerical simulations based on the discrete dipole approximation (DDA), SERS enhancement originates from high electromagnetic fields (hot spots) localized both inside and outside individual NWs. In addition, far-field intercoupling effects between NWs have been observed experimentally in arrays with subwavelength pitch sizes. We show that the SERS enhancement factors can also be tuned and optimized by adjusting the geometry of NWs. PMID- 18507400 TI - Differential-phase surface plasmon resonance biosensor. AB - In this paper, a novel differential-phase-sensitive surface plasmon resonance biosensor (DP-SPRB) is proposed and developed, in which a two-frequency laser is integrated with a differential amplifier in order to analytically convert the phase modulation into amplitude modulation. With the use of the conventional envelope detection technique, the differential phase is precisely decoded in real time in terms of the demodulated amplitude. In order to verify high detection sensitivity of the DP-SPRB, a sucrose-water solution and glycerin-water solution at low concentrations were both tested, and the experimental results confirm that the detection sensitivity on wt % concentration of the sucrose solution is 0.00001%. Moreover, the real-time monitoring mouse IgG/antimouse IgG interaction shows the minimum concentration of mouse IgG to be at 10 fg/mL. To our knowledge, this is the highest sensitivity ever measured by a surface plasmon resonance biosensor. However, because of the limited dynamic range of DP-SPRB, it can only apply to biomolecule interactions at extremely low concentration. PMID- 18507401 TI - Integration or segregation: how do molecules behave at oil/water interfaces? AB - It has been over 250 years since Benjamin Franklin, fascinated with the wave stilling effect of oil on water, performed his famous oil-drop experiments; nevertheless, the behavior of water molecules adjacent to hydrophobic surfaces continues to fascinate today. In the 18th century, the calming of the seas seemed the most pertinent application of such knowledge; today, we understand that oil on-water phenomena underlie a range of important chemical, physical, and biological processes, including micelle and membrane formation, protein folding, chemical separation, oil extraction, nanoparticle formation, and interfacial polymerization. Beyond classical experiments of the oil-water interface, recent interest has focused on deriving a molecular-level picture of this interface or, more generally, of water molecules positioned next to any hydrophobic surface. This Account summarizes more than a decade's work from our laboratories aimed at understanding the nature of the hydrogen bonding occurring between water and a series of organic liquids in contact. Although the common perception is that water molecules and oil molecules positioned at the interface between the immiscible liquids want nothing to do with one another, we have found that weak interactions between these hydrophilic and hydrophobic molecules lead to interesting interfacial behavior, including highly oriented water molecules and layering of the organic medium that extends several molecular layers deep into the bulk organic liquid. For some organic liquids, penetration of oriented water into the organic layer is also apparent, facilitated by molecular interactions established at the molecularly thin region of first contact between the two liquids. The studies involve a combined experimental and computational approach. The primary experimental tool that we have used is vibrational sum frequency spectroscopy (VSFS), a powerful surface-specific vibrational spectroscopic method for measuring the molecular structures of aqueous surfaces. We have compared the results of these spectroscopic studies with our calculated VSF spectra derived from population densities and orientational distributions determined through molecular dynamics (MD) simulations. This combination of experiment and theory provides a powerful opportunity to advance our understanding of molecular processes at aqueous interfaces while also allowing us to test the validity of various molecular models commonly used to describe molecular structure and interactions at such interfaces. PMID- 18507402 TI - A combinatorial polymer library approach yields insight into nonviral gene delivery. AB - The potential of gene therapy to benefit human health is tremendous because almost all human diseases have a genetic component, from untreatable monogenic disorders to cancer and heart disease. Unfortunately, a method for gene therapy that is both effective and safe has remained elusive. It has been said that "there are only three problems in gene therapy - delivery, delivery, and delivery." (quote from I. M. Verma in Jaroff, L. TIME, 1999; Jan 11). This Account describes an alternative strategy to viral gene delivery: the design of biodegradable polymers that are able to deliver DNA like a synthetic virus. Using high-throughput synthesis and screening techniques, we have created libraries of over 2000 structurally unique poly(beta-amino esters) (PBAEs). PBAEs are formed by the conjugate addition of amines to diacrylates. These biomaterials are promising for nonviral gene delivery due to their ability to condense plasmid DNA into small and stable nanoparticles and their ability to promote cellular uptake and endosomal escape. Our laboratory has iteratively improved PBAE nanoparticles through polymer end modifications and nanoparticle coatings. Lead PBAEs have high gene delivery efficacy and low cytotoxicity both in vitro and in vivo. Certain polymer structural characteristics are important for effective gene delivery. The best PBAEs are linear polymers of ~10 kDa that contain hydroxyl side chains and primary amine end groups. These polymers bind DNA to form nanoparticles that are small (<200 nm) and stable and have near-neutral zeta potential in the presence of serum-containing media. Lead PBAEs also contain tertiary amines that can buffer the low pH environment of endosomes and facilitate escape of polymer/DNA particles into the cytoplasm. Diamine end-modified 1,4-butanediol diacrylate-co-5 amino-1-pentanol polymers (C32) bind DNA more tightly and form smaller nanoparticles than other PBAEs. These nanoparticles also have higher cellular uptake and the best gene expression of all gene delivery polymers in the library. These polymers are more effective for gene delivery than top commercially available nonviral vectors including jet-PEI and Lipofectamine 2000 and are comparable to adenovirus for in vitro gene delivery to human primary cells. In vivo, these PBAE/DNA particles are promising as cancer therapeutics. This Account summarizes the results of our laboratory in using a combinatorial polymer library approach to elucidate polymer structure/function relationships and enable the development of polymeric gene delivery nanoparticles with viral-like efficacy. PMID- 18507403 TI - Crops: a green approach toward self-assembled soft materials. AB - To date, a wide range of industrial materials such as solvents, fuels, synthetic fibers, and chemical products are being manufactured from petroleum resources. However, rapid depletion of fossil and petroleum resources is encouraging current and future chemists to orient their research toward designing safer chemicals, products, and processes from renewable feedstock with an increased awareness of environmental and industrial impact. Advances in genetics, biotechnology, process chemistry, and engineering are leading to a new manufacturing concept for converting renewable biomass to valuable fuels and products, generally known as the biorefinery concept. The swift integration of crop-based materials synthesis and biorefinery manufacturing technologies offers the potential for new advances in sustainable energy alternatives and biomaterials that will lead to a new manufacturing paradigm. This Account presents a novel and emerging concept of generating various forms of soft materials from crops (an alternate feedstock). In future research, developing biobased soft materials will be a fascinating yet demanding practice, which will have direct impact on industrial applications as an economically viable alternative. Here we discuss some remarkable examples of glycolipids generated from industrial byproducts such as cashew nut shell liquid, which upon self-assembly produced soft nanoarchitectures including lipid nanotubes, twisted/helical nanofibers, low-molecular-weight gels, and liquid crystals. Synthetic methods applied to a "chiral pool" of carbohydrates using the selectivity of enzyme catalysis yield amphiphilic products derived from biobased feedstock including amygdalin, trehalose, and vitamin C. This has been achieved with a lipase-mediated regioselective synthetic procedure to obtain such amphiphiles in quantitative yields. Amygdalin amphiphiles showed unique gelation behavior in a broad range of solvents such as nonpolar hexanes to polar aqueous solutions. Importantly, an enzyme triggered drug-delivery model for hydrophobic drugs was demonstrated by using these supramolecularly assembled hydrogels. Following a similar biocatalytic approach, vitamin C amphiphiles were synthesized with different hydrocarbon chain lengths, and their ability to self-assemble into molecular gels and liquid crystals has been studied in detail. Such biobased soft materials were successfully used to develop novel organic-inorganic hybrid materials by in situ synthesis of metal nanoparticles. The self-assembled soft materials were characterized by several spectroscopic techniques, UV-visible, infrared, and fluorescence spectrophotometers, as well as microscopic methods including polarized optical, confocal, scanning, and transmission electron microscopes, and thermal analysis. The molecular packing of the hierarchically assembled bilayer membranes was fully elucidated by X-ray analysis. We envision that the results summarized in this Account will encourage interdisciplinary collaboration between scientists in the fields of organic synthesis, soft materials research, and green chemistry to develop functional materials from underutilized crop-based renewable feedstock, with innovation driven both by material needs and environmentally benign design principles. PMID- 18507404 TI - The interface energetics of self-assembled monolayers on metals. AB - Self-assembled monolayers (SAMs) of organic molecules generally modify the surface properties when covalently linked to substrates. In organic electronics, SAMs are used to fine-tune the work functions of inorganic electrodes, thereby minimizing the energy barriers for injection or extraction of charge carriers into or out of an active organic layer; a detailed understanding of the interface energetics on an atomistic scale is required to design improved interfaces. In the field of molecular electronics, the SAM itself (or, in some cases, one or a few molecules) carries the entire device functionality; the interface then essentially becomes the device and the alignment of the molecular energy levels with those of the electrodes defines the overall charge-transport characteristics. This Account provides a review of recent theoretical studies of the interface energetics for SAMs of pi-conjugated molecules covalently linked to noble metal surfaces. After a brief description of the electrostatics of dipole layers at metal/molecule interfaces, the results of density functional theory calculations are discussed for SAMs of representative conjugated thiols on Au(111). Particular emphasis is placed on the modification of the work function of the clean metal surface upon SAM formation, the alignment of the energy levels within the SAM with the metal Fermi level, and the connection between these two quantities. To simplify the discussion, we partition the description of the metal/SAM system into two parts by considering first an isolated free-standing layer of molecules and then the system obtained after molecule-metal bond formation. From an electrostatic standpoint, both the isolated monolayer and the metal-molecule bonds can be cast in the form of dipole layers, which lead to steps in the electrostatic potential energy at the interface. While the step due to the isolated molecular layer impacts only the work function of the SAM-covered surface, the step arising from the bond formation influences both the work function and the alignment of the electronic levels in the SAM with respect to the metal Fermi energy. Interestingly, headgroup substitutions at the far ends of the molecules forming the SAM are electrostatically decoupled from the metal thiol interface in densely packed SAMs; as a result, the nature of these substituents and the binding chemistry between the metal and the molecules are two largely unrelated handles with which to independently tune the work function and the level alignment. The establishment of a comprehensive atomistic picture regarding the impact of the individual components of a SAM on the interface energetics at metal/organic junctions paves the way for clear guidelines to design improved functional interfaces in organic and molecular electronics. PMID- 18507405 TI - Mesostructured spheres of organic/inorganic hybrid from gelable block copolymers and arched nano-objects thereof. AB - Mesostructured microspheres formed by aerosol-assisted self-assembly of a gelable block copolymer, poly(3-(triethoxysilyl)propyl methacrylate)- block-polystyrene (PTEPM -b-PS), were studied by a combination of small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). When the copolymer composition was changed, the spheres with different internal patterns, such as onion-like lamella and curved cylinder, were obtained. Through a self-gelation process of PTEPM domains, novel organic/inorganic hybrid spheres with an internal tunable patterned structure were prepared. Since only PTEMP domains were cross-linked, the hybrid spheres could be further disintegrated by dispersion in a good solvent of PS. As a result, novel organic/inorganic hybrid nano-objects such as arched plates and cylinders were prepared. PMID- 18507406 TI - Quantitative measurement of nanoparticle halo formation around colloidal microspheres in binary mixtures. AB - A new colloidal stabilization mechanism, known as nanoparticle "haloing" (Tohver, V.; Smay, J. E.; Braem, A.; Braun, P. V.; Lewis, J. A. Proc. Natl. Acad. Sci. U.S.A. 2001, 98, (16), 8950-8954), has been predicted theoretically and inferred experimentally in microsphere-nanoparticle mixtures that possess high charge and size asymmetry. The term "halo" implies the existence of a nonzero separation distance between the highly charged nanoparticles and the negligibly charged microspheres that they surround. By means of ultrasmall-angle X-ray scattering, we have quantified the microsphere-nanoparticle separation distance as well as the number of nanoparticles and their lateral separation distance within the self organized halos that form in these binary mixtures. PMID- 18507407 TI - Structure and conductance of aromatic and aliphatic dithioacetamide monolayers on Au(111). AB - The structure and electrical properties of self-assembled monolayers of cyclic aromatic and aliphatic dithioacetamides (1,4-bis(mercaptoacetamido)benzene and 1,4-bis(mercaptoacetamido)cyclohexane) and of mixed dithioacetamide/alkanethiol monolayers are characterized by X-ray photoelectron spectroscopy (XPS), scanning tunneling microscopy (STM) and contact angle goniometry. Both dithioacetamides are found to pack densely on Au(111), however the monolayers are poorly ordered as a result of hydrogen bond formation between the amide groups. The coassembly and the insertion method are compared for the formation of mixed dithioacetamide/alkanethiol monolayers. By coassembly, islands of dithioacetamides in a dodecanethiol matrix can only be obtained at a low dithioacetamide/dodecanethiol concentration ratio in solution (1/10) and by thermal annealing of the resulting monolayers. Small and well defined dithioacetamide domains are realized by insertion of dithioacetamides into defect sites of closely packed octanethiol monolayers. These domains are used to determine the molecular conductance by means of STM height profiles and molecular lengths resulting from density functional theory (DFT) calculations. The difference in the tunneling decay constant beta measured for aromatic dithioacetamides (beta = 0.74-0.76/A) and for aliphatic dithioacetamides (beta = 0.84-0.91/A) highlights the influence of the conjugation within the cyclic core on molecular conductance. PMID- 18507408 TI - Arrest of fluid demixing by nanoparticles: a computer simulation study. AB - We use lattice Boltzmann simulations to investigate the formation of arrested structures upon demixing of a binary solvent containing neutrally wetting colloidal particles. Previous simulations for symmetric fluid quenches pointed to the formation of "bijels": bicontinuous interfacially jammed emulsion gels. These should be created when a glassy monolayer of particles forms at the fluid-fluid interface, arresting further demixing and rigidifying the structure. Experimental work has broadly confirmed this scenario, but it shows that bijels can also be formed in volumetrically asymmetric quenches. Here, we present new simulation results for such quenches, compare these to the symmetric case, and find a crossover to an arrested droplet phase at strong asymmetry. We then make extensive new analyses of the postarrest dynamics in our simulated bijel and droplet structures, on time scales comparable to the Brownian time for colloid motion. Our results suggest that, on these intermediate time scales, the effective activation barrier to ejection of particles from the fluid-fluid interface is smaller by at least 2 orders of magnitude than the corresponding barrier for an isolated particle on a flat interface. PMID- 18507409 TI - Self-consistent field modeling of adsorption from polymer/surfactant mixtures. AB - We report on the development of a self-consistent field model that describes the competitive adsorption of nonionic alkyl-(ethylene oxide) surfactants and nonionic polymer poly(ethylene oxide) (PEO) from aqueous solutions onto silica. The model explicitly describes the response to the pH and the ionic strength. On an inorganic oxide surface such as silica, the dissociation of the surface depends on the pH. However, salt ions can screen charges on the surface, and hence, the number of dissociated groups also depends on the ionic strength. Furthermore, the solvent quality for the EO groups is a function of the ionic strength. Using our model, we can compute bulk parameters such as the average size of the polymer coil and the surfactant CMC. We can make predictions on the adsorption behavior of either polymers or surfactants, and we have made adsorption isotherms, i.e., calculated the relationship between the surface excess and its corresponding bulk concentration. When we add both polymer and surfactant to our mixture, we can find a surfactant concentration (or, more precisely, a surfactant chemical potential) below which only the polymer will adsorb and above which only the surfactant will adsorb. The corresponding surfactant concentration is called the CSAC. In a first-order approximation, the surfactant chemical potential has the CMC as its upper bound. We can find conditions for which CMC < CSAC . This implies that the chemical potential that the surfactant needs to adsorb is higher than its maximum chemical potential, and hence, the surfactant will not adsorb. One of the main goals of our model is to understand the experimental data from one of our previous articles. We managed to explain most, but unfortunately not all, of the experimental trends. At the end of the article we discuss the possibilities for improving the model. PMID- 18507410 TI - Multifunctional mixed SAMs that promote both cell adhesion and noncovalent DNA immobilization. AB - The ability of DNA strands to influence cellular gene expression directly and to bind with high affinity and specificity to other biological molecules (e.g., proteins and target DNA strands) makes them a potentially attractive component of cell culture substrates. On the basis of the potential importance of immobilized DNA in cell culture and the well-defined characteristics of alkanethiol self assembled monolayers (SAMs), the current study was designed to create multifunctional SAMs upon which cell adhesion and DNA immobilization can be independently modulated. The approach immobilizes the fibronectin-derived cell adhesion ligand Arg-Gly-Asp-Ser-Pro (RGDSP) using carbodiimide activation chemistry and immobilizes DNA strands on the same surface via cDNA-DNA interactions. The surface density of hexanethiol-terminated DNA strands on alkanethiol monolayers (30.2-69.2 pmol/cm2) was controlled using a backfill method, and specific target DNA binding on cDNA-containing SAMs was regulated by varying the soluble target DNA concentration and buffer characteristics. The fibronectin-derived cell adhesion ligand GGRGDSP was covalently linked to carboxylate groups on DNA-containing SAM substrates, and peptide density was proportional to the amount of carboxylate present during SAM preparation. C166 GFP endothelial cells attached and spread on mixed SAM substrates and cell adhesion and spreading were specifically mediated by the immobilized GGRGDSP peptide. The ability to control the characteristics of noncovalent DNA immobilization and cell adhesion on a cell culture substrate suggests that these mixed SAMs could be a useful platform for studying the interaction between cells and DNA. PMID- 18507411 TI - Determination of the adsorption free energy for peptide-surface interactions by SPR spectroscopy. AB - To understand and predict protein adsorption behavior, we must first understand the fundamental interactions between the functional groups presented by the amino acid residues making up a protein and the functional groups presented by the surface. Limited quantitative information is available, however, on these types of submolecular interactions. The objective of this study was therefore to develop a reliable method to determine the standard state adsorption free energy (delta Go ads) of amino acid residue-surface interactions using surface plasma resonance (SPR) spectroscopy. Two problems are commonly encountered when using SPR for peptide adsorption studies: the need to account for "bulk-shift" effects and the influence of peptide-peptide interactions at the surface. Bulk-shift effects represent the contribution of the bulk solute concentration to the SPR response that occurs in addition to the response due to adsorption. Peptide peptide interactions, which are assumed to be zero for Langmuir adsorption, can greatly skew the isotherm shape and result in erroneous calculated values of delta Go ads. To address these issues, we have developed a new approach for the determination of delta Go ads using SPR that is based on the chemical potential. In this article, we present the development of this new approach and its application for the calculation of delta Go ads for a set of peptide-surface systems where the peptide has a host-guest amino acid sequence of TGTG-X-GTGT (where G and T are glycine and threonine residues and X represents a variable residue) and the surface consists of alkanethiol self-assembled monolayers (SAMs) with methyl (CH 3) and hydroxyl (OH) functionality. This new approach enables bulk-shift effects to be directly determined from the raw SPR versus peptide concentration data plots and the influence of peptide-peptide interaction effects to be minimized, thus providing a very straightforward and accurate method for the determination of delta Go ads for peptide adsorption. Further studies are underway to characterize delta Go ads for a large library of peptide-SAM combinations. PMID- 18507412 TI - The quartz crystal microbalance: a new tool for the investigation of the bioadhesion of diatoms to surfaces of differing surface energies. AB - Diatoms are a major component of the biofoul layer found on modern low-surface energy, 'foul release' coatings. While diatoms adhere more strongly to hydrophobic, as opposed to hydrophilic, surfaces, surprisingly little is known of the chemical composition of their adhesives. Even less is known about the underlying processes that characterize the interaction between the adhesive and a given surface, including those of differing wettability. Using the quartz crystal microbalance with dissipation monitoring (QCM-D), we examined differences in the viscoelastic properties of the extracellular adhesives produced by the marine diatoms Amphora coffeaeformis Cleve and Craspedostauros australis Cox interacting with surfaces of differing wettability; 11-mercaptoundecanoic acid (MUA) that is hydrophilic and 1-undecanethiol (UDT) that is hydrophobic. While the overall delta f/delta D ratios were slightly different, the trends were the same for both diatom species, with the layer secreted upon UDT to be more viscoelastic and far more consistent over several experiments, compared to that on MUA which was less viscoelastic and demonstrated far more variability between experiments. While the nature of the parameter shifts for C. australis were the same for both surfaces, A. coffeaeformis cells settling upon UDT illustrated significant positive f and D shifts during the initial stages of cell settlement and adhesion to the surface. Further experiments revealed the parameter shifts to occur only during the initial adhesion of cells upon the pristine virgin UDT surface. The mechanism behind these parameter responses was isolated to the actin-myosin/adhesion complex (AC), using the myosin inhibitor 2,3-butanedione 2-monoxime (BDM) to remove the cells ability to 'pull' on adhesive strands emanating from the cell raphe. The observations made herein have revealed that adhesives secreted by fouling diatoms differ significantly in their interaction with surfaces depending on their wettability, as well as illustrating the unique mechanics behind the adhesion of A. coffeaeformis upon hydrophobic surfaces, a mechanism that may contribute significantly to the cells success in colonizing hydrophobic surfaces. PMID- 18507413 TI - Kinetics of solute adsorption at solid/solution interfaces: on the special features of the initial adsorption kinetics. AB - The features of the initial adsorption kinetics monitored at short adsorption times are investigated. It is shown that the concave character of the square-root dependence on time may be due to a combined effect of the rate of surface reaction and that of the transport from the bulk to the surface. That effect causes the appearance of a certain subsurface region close to the surface, where the concentration of the sorbate is different from that in the bulk phase. For the purpose of illustration, the initial parts of the kinetic isotherms are analyzed for the RY/F-400 system already studied in our previous paper. PMID- 18507414 TI - CO2 capture by polyethylenimine-modified fibrous adsorbent. AB - This work focuses on developing a novel adsorbent for CO2 capture, by coating polyethylenimine (PEI) on glass fiber matrix and using epichlorohydrin (ECH) as cross-linking agent. The physicochemical properties of the fibrous adsorbent were characterized. The CO2 adsorption capacity was evaluated. Factors that affect the adsorption capacity of the fibrous adsorbent were studied. The experimental results show that this fibrous PEI adsorbent exhibits a much higher adsorption capacity for CO2 compared with another PEI fiber prepared in our previous work, which employed epoxy resin as the cross-linking agent. A CO2 adsorption capacity as high as 4.12 mmol CO2/g of adsorbent was obtained for this fibrous PEI adsorbent at 30 degrees C, equal to 13.56 mmol CO2/g of PEI, with a PEI/ECH ratio of 20:1. The adsorbent can be completely regenerated at 120 degrees C. PMID- 18507415 TI - On pattern transfer in replica molding. AB - Nano- and micromolding of elastic materials produces smoothed replicas of the mold structures. This limits the technique's resolution. Here we identified surface tension as the cause of smoothing and derived explicit equations for calculating molded feature shapes. The characteristic length scale for smoothing is given by the ratio of the interface tension to Young's modulus of the molded material. This approach offers the possibility to correct for the smoothing caused by surface tension during mold design. Moreover, it can be exploited to measure interface tension. PMID- 18507416 TI - Immobilized chemoattractant peptides mediate adhesion and distinct calcium dependent cell signaling in human neutrophils. AB - Chemotaxis is the stimulated directional migration of cells in response to chemotactic factors, manifested for instance during leukocyte interaction with chemoattractants in inflammation. The N-formyl-Met-Leu-Phe (fMLF) bacterial peptide family is particularly potent in attracting and activating neutrophilic granulocytes. To accomplish defined circumstances for recruitment and activation of cells, we fabricated semitransparent gold-coated glass coverslips functionalized with chemoattractant fMLF receptor peptide agonist analogues. Peptides based on a common leading four-amino-acid sequence Gly-Gly-Gly-Cys were thus coupled to two potent fMLF receptor agonists, N-formyl-Tyr-Nle-Phe-Leu-Nle Gly-Gly-Gly-Cys and N-formyl-Met-Leu-Phe-Gly-Gly-Gly-Cys, and a formylated control peptide, N-formyl-Gly-Gly-Gly-Cys. They were anchored via the SH group of Cys either directly to the gold surface or a mixed self-assembled monolayer composed of maleimide- and hydroxyl-terminated oligo(ethylene glycol) alkyldisulfides. The overall peptide immobilization procedure was characterized with ellipsometry, contact angle measurement, and infrared spectroscopy. When exposed to granulocytes, the agonist surface rapidly recruited neutrophils and the cells responded with extensive spreading and intracellular calcium transients within minutes. The reference peptide generated no such activation, and the cells maintained a more spherical morphology, suggesting that we have been able to immobilize chemoattractant receptor agonist peptides with retained bioactivity. This is a crucial step in designing surfaces with specific effects on cellular behavior. PMID- 18507417 TI - Grafting acrylic polymers from flat nickel and copper surfaces by surface initiated atom transfer radical polymerization. AB - Acrylic polymers, including poly(methyl methacrylate), poly(2,2,2-trifluoroethyl methacrylate), poly( N,N'-dimethyaminoethyl methacrylate), and poly(2 hydroxyethyl methacrylate) were grafted from flat nickel and copper surfaces through surface-initiated atom transfer radical polymerization (ATRP). For the nickel system, there was a linear relationship between polymer layer thickness and monomer conversion or molecular weight of "free" polymers. The thickness of the polymer brush films was greater than 80 nm after 6 h of reaction time. The grafting density was estimated to be 0.40 chains/nm2. The "living" chain ends of grafted polymers were still active and initiated the growth of a second block of polymer. Block copolymer brushes with different block sequences were successfully prepared. The experimental surface chemical compositions as measured by X-ray photoelectron spectroscopy agreed very well with their theoretical values. Water contact angle measurements further confirmed the successful grafting of polymers from nickel and copper surfaces. The surface morphologies of all samples were studied by atomic force microscopy. This study provided a novel approach to prepare stable functional polymer coatings on reactive metal surfaces. PMID- 18507418 TI - Structure and dynamics of micelles and cubic phase structures with ethoxylated phytosterol surfactant in water. AB - The self-assembly of a sterol ethoxylate surfactant with 30 oxyethylene units in water was studied by 1H NMR self-diffusion measurements in a wide concentration range in the micellar region (0-25 wt %). The data showed that the surfactant aggregates do not interact by hard sphere interactions but rather a strong concentration dependence of the diffusion coefficient was noted which was explained by polymer scaling theory. In the cubic phase (30-65 wt %), the self diffusion data from water, from surfactant, and from free polyoxyethylene suggest spherical micelles, although water diffusion was much restricted due to binding to the surfactant headgroup. From X-ray measurements in the cubic phase, the unit cell size was calculated, and together with surfactant self-diffusion measurements the exchange dynamics between free and aggregated surfactant was obtained. PMID- 18507419 TI - Adsorption of low charge density polyelectrolytes to an oppositely charged porous substrate. AB - The adsorption behavior of a low charge density cationic polyelectrolyte to cellulosic fibers has been studied. Cationic dextran served as a model polyelectrolyte, as it can be prepared over a range in molecular mass and charge density. The adsorption behavior of the cationic dextran was measured in electrolyte-free conditions using polyelectrolyte titration techniques. By fluorescent labeling the cationic dextran, the extent to which adsorption occurs inside the porous structure was further determined by fluorescent confocal laser scanning microscopy. Cationic dextran having a sufficiently low charge density adsorbed into the pores, although the extent the cationic dextran adsorbed was governed by the molecular mass. The adsorption behavior of the cationic dextran was also studied in various electrolyte concentrations. The adsorbed mass monotonically decreased with increasing electrolyte, as the electrostatic interaction with the substrate was more effectively screened. This behavior also suggests that the interactions between adsorbed polyelectrolyte chains, i.e. lateral correlation effects, are negligible for low charge density polyelectrolytes. Finally, the effect of having a preadsorbed layer of cationic dextran on the adsorption behavior was determined in electrolyte-free conditions using fluorescent double staining techniques. The preadsorbed cationic dextran had almost no effect on the adsorption of low molecular mass fractions. Low molecular mass fractions directly adsorbed into the pore structure, as opposed to adsorbing to a free surface and diffusing into the pores. It was also shown that cationic dextran can be selectively adsorbed to different locations, such that the surface of a porous substrate can be treated uniquely from the bulk. PMID- 18507420 TI - Creation of functional membranes using polyelectrolyte multilayers and polymer brushes. AB - Over the last 15 years, the layer-by-layer deposition of polyelectrolytes and the growth of polymer brushes from surfaces have become established techniques for the formation of a wide range of thin films. This article discusses the use of these techniques in creating the skin layer of nanofiltration or gas-separation membranes and in functionalizing the interior of membranes for protein adsorption or catalysis. In the case of separation membranes for nanofiltration, the minimal thickness of layer-by-layer films allows for high flux, and the wide range of available polyelectrolytes that can form these films permits the tailoring of membranes for separations such as water softening, the reduction of F (-) concentrations, and the removal of dyes from wastewater. For gas separation, polymers grown from surfaces are more attractive than layer-by-layer coatings because most polyelectrolyte films are not highly gas-selective. Cross-linked poly(ethylene glycol dimethacrylate) films grown from porous alumina exhibit CO(2)/CH(4) selectivities of around 20, and the careful selection of monomers should further improve the selectivity of similar membranes. Both layer-by-layer methods and polymer brushes can also be employed to modify the interior of membranes, and we have utilized these techniques to create catalysts, antibody arrays in membranes, and membrane absorbers for protein purification. Polymer brushes are particularly attractive because they allow the absorption of multilayers of protein to yield membranes with binding capacities as high as 150 mg protein/cm(3). Some challenges in the practical implementation of these systems, such as the economical formation of membranes using highly permeable polymeric supports, and future directions in research on membrane modification with multilayer films and polymer brushes are also discussed herein. PMID- 18507421 TI - Metallic LiMo3Se3 nanowire film sensors for electrical detection of metal ions in water. AB - LiMo 3Se 3 nanowire film sensors were fabricated by drop-coating a 0.05% (mass) aqueous nanowire solution onto microfabricated indium tin oxide electrode pairs. According to scanning electron microscopy (SEM) and atomic force microscopy (AFM), the films are made of a dense network of 3-7 nm thick nanowire bundles. Immersion of the films in 1.0 M aqueous solutions of group 1 or 2 element halides or of Zn(II), Mn(II), Fe(II), or Co(II) chlorides results in an increase of the electrical resistance of the films. The resistance change is always positive and reaches up to 9% of the base resistance of the films. It occurs over the course of 30-240 s, and it is reversible for monovalent ions and partially reversible for divalent ions. The signal depends on the concentration of the electrolyte and on the size and charge of the metal cation. Anions do not play a significant role, presumably, because they are repelled by the negatively charged nanowire strands. The magnitude of the electrical response and its sign suggest that it is due to analyte-induced scattering of conduction electrons in the nanowires. An ion-induced field effect can be excluded based on gated conductance measurements of the nanowire films. PMID- 18507422 TI - Adsorption of the protein bovine serum albumin in a planar poly(acrylic acid) brush layer as measured by optical reflectometry. AB - The adsorption of bovine serum albumin (BSA) in a planar poly(acrylic acid) (PAA) brush layer has been studied by fixed-angle optical reflectometry. The influence of polymer length, grafting density, and salt concentration is studied as a function of pH. The results are compared with predictions of an analytical polyelectrolyte brush model, which incorporates charge regulation and excluded volume interactions. A maximum in adsorption is found near the point of zero charge (pzc) of the protein. At the maximum, BSA accumulates in a PAA brush to at least 30 vol %. Substantial adsorption continues above the pzc, that is, in the pH range where a net negatively charged protein adsorbs into a negatively charged brush layer, up to a critical pH value. This critical pH value decreases with increasing ionic strength. The adsorbed amount increases strongly with both increasing PAA chain length and increasing grafting density. Experimental data compare well with the analytical model without having to include a nonhomogeneous charge distribution on the protein surface. Instead, charge regulation, which implies that the protein adjusts its charge due to the negative electrostatic potential in the brush, plays an important role in the interpretation of the adsorbed amounts. Together with nonelectrostatic interactions, it explains the significant protein adsorption above the pzc. PMID- 18507423 TI - Comparison between inhomogeneous adsorption of charged surfactants on air-water and on solid-water interfaces by self-consistent field theory. AB - We use a realistic molecular model to study the interfacial behavior of hydrocarbon sulfate surfactants within a self-consistent field model and consider the adsorption both at the air-water interface and at a hydrophobic solid-water interface. We focus on the structural properties of the hemimicelles at the critical interface aggregation concentration (CIAC) for the air-water system and the critical surface aggregation concentration (CSAC) for the solid-water system. The major difference between the two systems is that the liquid interface is penetrable but the solid surface is intrinsically impenetrable for the molecular species. At the LG interface the hemimicelles have a lens shape with their centers of mass positioned slightly toward the aqueous side and feature an aspect ratio of approximately 2, with the long dimension parallel to the interface. Hemimicelle formation occurs below a critical (interfacial) area per molecule and above a critical surface pressure depending on tail length and ionic strength. Hemimicelles are not expected at air-water interfaces for a surfactant with a tail length ( t) lower than 15 CH2 units. In contrast, at a hydrophobic solid the formation of laterally inhomogeneous micelles even takes place for surfactants with the tail length as short as t = 12. This difference is attributed to the screening of the lateral interactions in the vapor phase. The shape of surface hemimicelles is caplike (or half-lens) with an aspect ratio lower than 2 and the long dimension parallel to the solid surface. The tail length, the ionic strength, the adsorption energies, and the surfactant concentration have an effect on the surface micelle properties such as the aggregation number and size and shape. PMID- 18507424 TI - Spatial control of protein binding on lipid bimembrane using photoeliminative linker. AB - Protein adsorption and dissociation on cell membrane surfaces is a topic of important study to reveal biological processes including signal transduction and protein trafficking. We demonstrated here the establishment of a mimic model system for the spatial control of protein adsorption/elimination on a lipid bimembrane using a photochemical technique. The novel photoeliminative linker that we synthesized here consists of three distinct components: a substrate (biotin), a photoeliminative group (4-(4-(1-hydroxyethyl)-2-methoxy-5 nitrophenoxy)butanoic acid), and a lipid bimembrane-adsorbent group (farnesyl). The photoeliminative linker was inserted on the entire surface of the lipid bimembrane and two-dimensionally eliminated by spatial UV irradiation onto the membrane to create a biotin pattern. A target protein, streptavidin was selectively immobilized on the patterned biotin, although it was almost not attached on the nonirradiated region. The streptavidin array was selectively dissociated by UV irradiation onto the entire membrane. PMID- 18507425 TI - Synthesis of nanoparticle-cored dendrimers by convergent dendritic functionalization of monolayer-protected nanoparticles. AB - This article presents a synthesis method for nanoparticle-cored dendrimers (NCDs), which have dendritic architectures around a monolayer-protected gold nanoparticle. The synthesis method is based on a strategy in which the synthesis of monolayer-protected nanoparticles is followed by adding dendrons on functionalized nanoparticles by a single coupling reaction. NMR spectroscopy, IR spectroscopy, and thermogravimetric analysis (TGA) characterizations confirmed the successful coupling reaction between dendrons with different generations ([G1], [G2], and [G3]) and COOH-functionalized nanoparticles ( approximately Au201L71). The dendrimer wedge density also could be controlled by reacting nanoparticles having different loading of COOH groups ( approximately 60 and approximately 10% COOH of the 71 ligands per gold nanoparticle) with functionalized dendrons. Transmission electron microscope results showed that this synthesis strategy maintains the average size of the nanoparticle core during dendron coupling reactions. This control over the composition and core size makes the systematic study of NCDs with different generations possible. The chemical stability of NCDs was found to be affected by dendron generation around the nanoparticle core. The current-potential response of NCD films on microelectrode arrays exhibited better electrical conductivity for NCDs with lower dendron generation. PMID- 18507426 TI - Late stage of the phase-separation process: coalescence-induced coalescence, gravitational sedimentation, and collective evaporation mechanisms. AB - We study the separation in the binary and ternary mixtures of the water/surfactant C12E5/polymer PEG system. The phase separation in the mixtures at late stages is governed by two distinct mechanisms: the coalescence-induced coalescence and the droplet evaporation mechanism. We show that when the coalescence-induced coalescence process is globally terminated in the sample consisting of a dense system of domains, another mechanism, which we call the collective droplet evaporation, starts to dominate. It manifests itself as a front of "evaporating" domains, which propagates at constant speed in the system. We show that the collective evaporation is induced by the gravitational drift of large droplets. PMID- 18507427 TI - Strategy for efficient site-specific FRET-dye labeling of ubiquitin. AB - To study conformational changes within a single protein molecule, sp-FRET (single pair fluorescence resonance energy transfer) is an important technique to provide distance information. However, incorporating donor and acceptor dyes into the same protein molecule is not an easy task. Here, we report a strategy for the efficient double-labeling of a protein on a solid support. An ubiquitin mutant with two Cys mutations, one with high solvent accessibility and the other with low solvent accessibility, was constructed. The protein was bound to magnetic beads and reacted with the dyes. The first dye reacted with the side-chain of the Cys with the high solvent accessibility and the second with the other Cys under partially denaturing conditions. Using this method, we can easily label two dyes in a site-specific way on ubiquitin with a satisfied yield. The labeling sites for donor and acceptor dyes can be easily swapped. PMID- 18507428 TI - Development of a thyroid hormone receptor targeting conjugate. AB - Molecular conjugates of hormone receptor-ligands with molecular probes or functional domains are finding diverse applications in chemical biology. Whereas many examples of hormone conjugates that target steroid hormone receptors have been reported, practical ligand conjugates that target the nuclear thyroid hormone receptor (TRbeta) are lacking. TR-targeting conjugate scaffolds based on the ligands GC-1 and NH-2 and the natural ligand triiodothyronine (T3) were synthesized and evaluated in vitro and in cellular assays. Whereas the T3 or GC-1 based conjugates did not bind TRbeta with high affinity, the NH-2 inspired fluorescein-conjugate JZ01 showed low nanomolar affinity for TRbeta and could be used as a nonradiometric probe for ligand binding. A related analogue JZ07 was a potent TR antagonist that is 13-fold selective for TRbeta over TRalpha. JZ01 localizes in the nuclei of TRbeta expressing cells and may serve as a prototype for other TR-targeting conjugates. PMID- 18507429 TI - A novel prosthetic group for site-selective labeling of peptides for positron emission tomography. AB - Efficient methodologies for the radiolabeling of peptides with [(18)F]fluoride are a prerequisite to enabling commercialization of peptide-containing radiotracers for positron emission tomography (PET) imaging. It was the purpose of this study to investigate a novel chemoselective ligation reaction comprising conjugation of an [(18)F]-N-methylaminooxy-containing prosthetic group to a functionalized peptide. Twelve derivatives of general formula R1-CO-NH-Lys-Gly Phe-Gly-Lys-OH were synthesized where R1 was selected from a short list of moieties anticipated to be reactive toward the N-methylaminooxy group. Conjugation reactions were initially carried out with nonradioactive precursors to assess, in a qualitative manner, their general suitability for PET chemistry with only the most promising pairings progressing to full radiochemical assessment. Best results were obtained for the ligation of O-[2-(2 [(18)F]fluoroethoxy)ethyl]-N-methyl-N-hydroxylamine 18 to the maleimidopropionyl Lys-Gly-Phe-Gly-Lys-OH precursor 10 in acetate buffer (pH 5) after 1 h at 70 degrees C. The non-decay-corrected isolated yield was calculated to be 8.5%. The most encouraging result was observed with the combination 18 and 4-(2 nitrovinyl)benzoyl-Lys-Gly-Phe-Gly-Lys-OH, 9, where the conjugation reaction proceeded rapidly to completion at 30 degrees C after only 5 min. The corresponding non-decay-corrected radiochemical yield for the isolated (18)F labeled product 27 was 12%. The preliminary results from this study demonstrate the considerable potential of this novel strategy for the radiolabeling of peptides. PMID- 18507430 TI - Structure-activity investigation on the gene transfection properties of cardiolipin mimicking gemini lipid analogues. AB - A structure-activity relationship has been explored on the gene transfection efficiencies of cardiolipin mimicking gemini lipid analogues upon variation of length and hydrophilicity of the spacer between the cationic ammonium headgroups and lipid hydrocarbon chain lengths. All the gemini lipids were found to be highly superior in gene transfer abilities as compared to their monomeric lipid and a related commercially available formulation. Pseudoglyceryl gemini lipids bearing an oxyethylene (-CH2-(CH2-O-CH2)m-CH2-) spacer were found to be superior gene transfecting agents as compared to those bearing polymethylene (-CH2)m-) spacers. The major characteristic feature of the present set of gemini lipids is their serum compatibility, which is most often the major hurdle in liposome mediated gene delivery. PMID- 18507431 TI - Induction of apoptosis in Jeko-1 mantle cell lymphoma cell line by resveratrol: a proteomic analysis. AB - Therapies for mantle cell lymphoma (MCL) are clinically unsatisfactory, and the search for effective drugs in vitro might foster the evaluation of their activity in vivo. We have investigated the effects of the polyphenolic compound resveratrol on the MCL cell line Jeko-1 using a combination of flow cytometry, Western blotting and two-dimensional electrophoresis to identify the molecules involved in the induction of apoptosis and cell growth regulation. We show that resveratrol induces apoptosis in Jeko-1 cells and modulates several key molecules, including cyclin D1 (CCND1), p53 (TP53), p21 (CDKN1A), BCL2, BAX, Bcl XL (BCL2L1), caspase 9 (CASP9) and p27 (CDKN1B). By high-resolution 2D-PAGE and nano-reverse phase-high performance liquid chromatography coupled with tandem mass spectrometry, we identified 32 differentially expressed proteins in response to resveratrol treatment that belong to important cell death related networks (including c-myc, NF-kappaB and the mitochondrial apoptotic pathway). These findings may improve the understanding of mechanisms mediating the pro-apoptotic effects of resveratrol on MCL cells, and form the basis for its potential use as a therapeutic agent. PMID- 18507433 TI - The mannose 6-phosphate glycoprotein proteome. AB - Most luminal lysosomal proteins are synthesized as precursors containing mannose 6-phosphate (Man6-P) and a number of recent studies have conducted affinity purification of Man6-P containing proteins as a step toward defining the composition of the lysosome. Approximately 60 known lysosomal proteins have been found in such studies as well as many other Man-6-P glycoproteins, some of which represent new lysosomal proteins. The latter are of considerable interest from cell-biological and biomedical perspectives, but differentiating between them and other proteins remains a significant challenge. The aim of this study was to conduct a global analysis of the mammalian Man6-P glycoproteome, implementing technical and biostatistical methods to aid in the discovery and validation of lysosomal candidates. We purified Man6-P glycoproteins from 17 individual rat tissues. To distinguish nonspecific contaminants (i.e., abundant or "sticky" proteins that are not fully removed during purification) from specifically purified proteins, we conducted a semiquantitative mass spectrometric comparison of protein levels in nonspecific mock eluates versus specific affinity chromatography eluates to identify those proteins that are specifically purified. We identified 60 known lysosomal proteins, representing nearly all that are currently known to contain Man-6-P. We also find 136 other proteins that are specifically purified but which are not known to have lysosomal function. This approach provides a list of candidate lysosomal proteins and also provides insights into the relative distribution of Man6-P glycoproteins. PMID- 18507432 TI - Vaccinia peptides eluted from HLA-DR1 isolated from virus-infected cells are recognized by CD4+ T cells from a vaccinated donor. AB - Class II MHC proteins bind peptides and present them to CD4 (+) T cells as part of the immune system's surveillance of bodily tissues for foreign and pathogenic material. Antigen processing and presentation pathways have been characterized in detail in normal cells, but there is little known about the actual viral peptides that are presented to CD4 (+) T cells that signal infection. In this study, two dimensional LC-MS/MS was used to identify vaccinia virus-derived peptides among the hundreds to thousands of peptide antigens bound to the human class II MHC protein HLA-DR1 on the surface of vaccinia virus-infected cells. The peptides, derived from the I6L, D6R, and A10L viral proteins, were 15 residues in length, bound efficiently to HLA-DR1 as synthetic peptides, and were recognized by vaccinia-specific CD4 (+) T cells obtained from an immunized donor. PMID- 18507434 TI - Benign and atypical meningioma metabolic signatures by high-resolution magic angle spinning molecular profiling. AB - Meningiomas are neoplasms that arise from the leptomeningeal covering of the brain and spinal cord, accounting for 15%-20% of CNS tumors. The WHO classifies meningiomas into three histological grades: benign, atypical, and anaplasic in accordance with the clinical prognosis. Atypical and anaplasic meningiomas tend to recur. Sometimes, meningiomas with histological diagnosis of benign meningioma show clinical characteristics of atypical meningioma. In this context, high resolution magic-angle spinning (HR-MAS) spectroscopy of intact tissue from brain tumor biopsies has shown great potential as a support diagnostic tool. In this work, we show differences between benign and atypical meningiomas in HR-MAS molecular profiles of meningioma biopsies. Metabolic differences between meningioma grades include changes in the levels of glutathione. Glutathione role in cancer is still unclear, as it may act both as protective and pathogenic factor. Glutamine and glutamate, which are related to glutathione metabolism and have been associated with tumor recurrence, are also increased in atypical meningiomas. Other metabolites associated with tumor malignancy that show statistically significant differences between benign and atypical meningiomas include phosphocholine and phosphoethanolamine. Overall, this work suggests that the additional information obtained by NMR metabolomics applied to biopsies of human meningiomas may be useful for assessing tumor grade and determining optimum treatment strategies. PMID- 18507435 TI - Understanding the growth rates of polymer cocrystallization in the binary mixtures of different chain lengths. AB - Polymer materials often contain a polydispersity of molecular lengths. We studied the linear growth rates of polymer lamellar crystals in the binary mixtures of different chain lengths by means of dynamic Monte Carlo simulations. Both chain lengths were chosen large enough to perform chain folding upon crystal growth but not very large to avoid the effect of chain entanglement in the bulk phase. We found that the crystal growth rates exhibit a linear dependence upon the compositions of mixtures. This linear relation implies that the overall crystal growth rates are integrated by the separate contributions of variable-length single polymers, supporting the model of intramolecular crystal nucleation. In each event of crystal growth of single polymers, long chains yield more crystallinity than short chains. This high efficiency explains higher crystal growth rates of long chains than that of short chains, and the explanation is quite different from the traditional view on the basis of their different melting points. In addition, with a partial release of sliding diffusion for crystal thickening, a new dependence of crystal growth rates occurs near the dilute end of long-chain compositions at high temperatures, which can be attributed to the preference of integer-number chain folding at the crystal growth front. The preferred fold lengths may vary with chain lengths and thus influence the crystal growth rates. PMID- 18507436 TI - Anion fractionation and reactivity at air/water:methanol interfaces. Implications for the origin of hofmeister effects. AB - Anions are selectively enriched in interfacial layers. This universal phenomenon, first identified in connection with protein precipitation 120 years ago, underlies fundamental processes. Its physical causes, however, remain conjectural. It has been speculated that the more polarizable anions should have larger affinities for air/liquid interfaces, and that their reactivities toward gaseous species would be affected by whether the liquid is capped by hydroxyl groups, as in water itself, or by hydrophobic layers of organic contaminants. These issues are particularly relevant to the composition and fate of atmospheric aerosols. Recently, we found that fractionation factors, f X (-) , of simple anions at the air/water interface increase exponentially with ion radius, a X (-) . In this paper, we report new experimental results on a set of anions that include the large PF 6 (-) and the highly polarizable IO 3 (-) species. A strict ln f X (-) proportional, variant a X (-) correlation is confirmed. Experiments performed in { x wH 2O + (1 - x w)MeOH} mixtures show that f X (-) is almost independent of x w. Furthermore, O 3(g) oxidizes I (-) at virtually identical rates on H 2O and MeOH. PMID- 18507437 TI - Identification and conformational study of stable radiation-induced defects in sucrose single crystals using density functional theory calculations of electron magnetic resonance parameters. AB - One of the major stable radiation-induced radicals in sucrose single crystals (radical T2) has been identified by means of density functional theory (DFT) calculations of electron magnetic resonance parameters. The radical is formed by a net glycosidic bond cleavage, giving rise to a glucose-centered radical with the major part of the spin density residing at the C 1 carbon atom. A concerted formation of a carbonyl group at the C 2 carbon accounts for the relatively small spin density at C 1 and the enhanced g factor anisotropy of the radical, both well-known properties of this radical from several previous experimental investigations. The experimentally determined and DFT calculated proton hyperfine coupling tensors agree very well on all accounts. The influence of the exact geometrical configuration of the radical and its environment on the tensors is explored in an attempt to explain the occurrence and characteristics of radical T3, another major species that is most likely another conformation of T2. No definitive conclusions with regard to the actual structure of T3 could be arrived at from this study. However, the results indicate that, most likely, T3 is identical in chemical structure to T2 and that changes in the orientation of neighboring hydroxy groups or changes in the configuration of the neighboring fructose ring can probably not account for the type and size of the discrepancies between T2 and T3. PMID- 18507438 TI - Origin of the absorption maxima of the photoactive yellow protein resolved via ab initio multiconfigurational methods. AB - We discuss the role of the protein in controlling the absorption spectra of photoactive yellow protein (PYP), the archetype xanthopsin photoreceptor, using quantum mechanics/molecular mechanics (QM/MM) methods based on ab initio multireference perturbation theory, combined with molecular dynamics (MD) simulations. It is shown that in order to get results in agreement with the experimental data, it is necessary to use a model that allows for a proper relaxation of the whole system and treats the states involved in the electronic spectrum in a balanced way, avoiding biased results due to the effect of nonrepresentative electrostatic interactions on the chromophore. PMID- 18507439 TI - A concise synthesis of (+/-)-cacalol. AB - A simple synthesis of the natural product cacalol has been developed that proceeds in seven steps and 21-25% overall yield. Ortho-lithiation of 4 methylanisole and alkylation with 5-iodo-1-pentene, followed by intramolecular Friedel-Crafts alkylation, gave 5-methoxy-1,8-dimethyltetralin. This compound was then formylated in the 6-position. Baeyer-Villiger oxidation and hydrolysis of the resulting formate gave 6-hydroxy-5-methoxy-1,8-dimethyltetralin. Alkylation of the phenolic hydroxyl group with chloroacetone followed by cyclodehydration gave cacalol methyl ether. Deprotection of this aryl methyl ether yielded cacalol. PMID- 18507440 TI - The case for the intermediacy of monomeric metaphosphate analogues during oxidation of H-phosphonothioate, H-phosphonodithioate, and H-phosphonoselenoate monoesters: mechanistic and synthetic studies. AB - Studies on the reaction of H-phosphonothioate, H-phosphonodithioate, and H phosphonoselenoate monoesters with iodine in the presence of a base led to identification of a unique oxidation pathway, which consists of the initial oxidation of the sulfur or selenium atom in these compounds, followed by oxidative elimination of hydrogen iodide to generate the corresponding metaphosphate analogues. The intermediacy of the latter species during oxidation of the investigated H-phosphonate monoester derivatives with iodine was supported by various diagnostic experiments. The scope and limitation of these oxidative transformations for the purpose of the synthesis of nucleoside phosphorothioate, nucleoside phosphorodithioate, and nucleoside phosphoroselenoate diesters was also investigated. PMID- 18507441 TI - Enantioselective syntheses of (-)- and (+)-monomorine I. AB - A concise enantioselective total synthesis of unnatural (-)-monomorine I has been achieved starting from lactam 2 in 54% overall yield. Natural (+)-monomorine I was also synthesized. PMID- 18507442 TI - Indium(III)-catalyzed tandem reaction with alkynylbenzaldehydes and alkynylanilines to heteroaromatic compounds. AB - Starting from ortho-alkynylbenzaldehydes and ortho-alkynylanilines, In(OTf)3 catalyzed synthesis of ring-condensed heteroaromatic compounds was developed via a domino intramolecular nucleophilic attack/intermolecular cycloaddition/dehydration reaction. PMID- 18507443 TI - Structure-property investigations of conjugated thiophenes fused onto a dehydro[14]annulene scaffold. AB - A series of 12 thieno-fused macrocycles based on the dehydro[14]annulene framework have been prepared. Studies have focused on the optical and electronic properties of the dehydrobenzothieno[14]annulenes (DBTAs) and dehydrothieno[14]annulenes (DTAs) utilizing NMR spectroscopy, UV-vis spectrophotometry, electrochemistry, and DFT computations. X-ray crystal structures were also obtained for two of the macrocycles. The structure-property relationships were found to vary significantly based on the relative orientation of the thiophenes. The stability, properties, and reactivity of these macrocycles were found to be more typical of dehydroannulenes rather than oligothiophenes. PMID- 18507444 TI - Enantioselective Pd-catalyzed alpha-arylation of N-Boc-pyrrolidine: the key to an efficient and practical synthesis of a glucokinase activator. AB - A short and practical synthesis of glucokinase activator 1 was achieved utilizing a convergent strategy involving S(N)Ar coupling of activated aryl fluoride 11 with hydroxypyridine 9. The key to the success of the synthesis was the development of a novel method for enantioselective formation of alpha arylpyrrolidines during the course of the project. In this method, (-)-sparteine mediated enantioselective lithiation of N-Boc-pyrrolidine was followed by in situ transmetalation to zinc and Pd-catalyzed coupling with aryl bromide 3, proceeding in 92% ee. This transformation allowed the preparation of compound 1 in a 31% overall yield over six steps. PMID- 18507445 TI - Chiral mono boronic acid as fluorescent enantioselective sensor for mono alpha hydroxyl carboxylic acids. AB - New mono boronic acid was found to be an enantioselective fluorescent chemosensor for mono alpha-hydroxyl carboxylic acids, such as mandelic acid and lactic acid. The chiral sensor shows lower background fluorescence, higher fluorescence enhancement, and enantioselective recognition kinetics toward mandelic acids and lactic acids. PMID- 18507446 TI - Synthesis and solid-state study of supramolecular host-guest assemblies: Bis[6 O,6-O'-(1,2:3,4-diisopropylidene-alpha-D-galactopyranosyl)thiophosphoryl] dichalcogenides. AB - A complementary approach for studying structural details of complex solid materials formed by symmetrical and unsymmetrical dichalcogenides, which employs both X-ray diffraction (XRD) and solid-state NMR (SS NMR), is presented. The new diagnostic technique allows reversible crystallographic space group change and very subtle distortion of host geometry to be followed during guest migration in the crystal lattice. Bis[6-O,6-O'-(1,2:3,4-diisopropylidene-alpha-D galactopyranosyl)]thiophosphoryl selenenyl sulfide, a representative of wheel-and axle host (WAAH) molecules, can be synthesized in the solid state by grinding and gentle heating of disulfide 1 and diselenide 2. Full characterization of disulfide 1 in the solid phase has been reported (J. Org. Chem. 1995, 60, 2549). In the current work, the synthesis and both XRD and SS NMR studies of the isostructural diselenide substrate 2 are presented. A (31)P cross polarization magic angle spinning experiment is employed to follow the progress of synthesis of selenenyl sulfide 3 in the solid state. It is concluded that selenenyl sulfide exists in equilibrium with disulfide and diselenide in a 1:1:1 ratio in both the liquid and the powdered solid. A mixture of isostructural dichalcogenides crystallized from different solvents form three-component host-guest inclusion complexes with columnar architecture. In the host-guest complex of diselenide 2 with toluene (space group C2), columns of host molecules are in parallel orientations along all the axes, whereas in the structures of diselenide 2 with propan-2-ol and propan-1-ol (space group P3 2), the columns of host molecules lay along the 3-fold symmetry axis. Thermal processes effecting structural changes in the host lattice and the kinetics of reversible guest molecule diffusion were investigated using SS NMR spectroscopy. Finally, the Se/S scrambling phenomenon and limitations in the X-ray structure refinement of organic compounds containing selenium and sulfur in chains are discussed. PMID- 18507447 TI - Indium-mediated 1,2,4,5-hexatetraen-3-ylation of 4-acetoxy-2-azetidinones and their applications to the Diels-Alder reactions for the synthesis of 2 azetidinone derivatives. AB - 4-Acetoxy-2-azetidinones reacted with organoindium reagent generated in situ from indium and 1,6-dibromo-2,4-hexadiyne in the presence of LiCl in DMF to selectively produce 2-azetidinones possessing 1,2,4,5-hexatetraen-3-yl group on the C4-position. The Diels-Alder reactions of 4-(1,2,4,5-hexatetraen-3-yl)-2 azetidinones with a variety of dienophiles and subsequent aromatizations afforded valuable functional group-substituted 2-azetidinones in good yields. PMID- 18507448 TI - Photochemistry of N-acetyl-, N-trifluoroacetyl-, N- mesyl-, and N tosyldibenzothiophene sulfilimines. AB - Time-resolved infrared (TRIR) spectroscopy, product studies, and computational methods were applied to the photolysis of sulfilimines derived from dibenzothiophene that were expected to release acetylnitrene, trifluoroacetylnitrene, mesylnitrene, and tosylnitrene. All three methods provided results for acetylnitrene consistent with literature precedent and analogous experiments with the benzoylnitrene precursor, i.e., that the ground state multiplicity is singlet. In contrast, product studies clearly indicate triplet reactivity for trifluoroacetylnitrene, though TRIR experiments were more ambiguous. Product studies suggest that these sulfilimines are superior sources for sulfonylnitrenes, which have triplet grounds states, to the corresponding azides, and computational studies shed light on the electronic structure of the nitrenes. PMID- 18507449 TI - Nondestructive replication of self-ordered nanoporous alumina membranes via cross linked polyacrylate nanofiber arrays. AB - Ordered nanofiber arrays are a promising material platform for artificial adhesive structures, tissue engineering, wound dressing, sensor arrays, and self cleaning surfaces. Their production via self-ordered porous alumina hard templates serving as shape-defining molds is well-established. However, their release requires the destruction of the hard templates, the fabrication of which is costly and time-consuming, by wet-chemical etching steps with acids or bases. We report the nondestructive mechanical extraction of arrays of cross-linked polyacrylate nanofibers from thus recyclable self-ordered nanoporous alumina hard templates. Silica replicas of the latter were synthesized using the extricated nanofiber arrays as secondary molds that could be mechanically detached from the molded material. The approach reported here, which can be combined with microstructuring, may pave the way for the high-throughput production of both functional nanofiber arrays and ordered nanoporous membranes consisting of a broad range of material systems. PMID- 18507450 TI - Infrared action spectroscopy of the OD stretch fundamental and overtone transitions of the DOOO radical. AB - The DOOO radical has been produced by three-body association between OD and O2 in a supersonic free-jet expansion and investigated using action spectroscopy, an IR UV double-resonance technique. Partially rotationally structured bands observed at 2635.06 and 5182.42 cm(-1) are assigned to the OD stretch fundamental (nu(OD)) and overtone (2nu(OD)), respectively, of the trans-DOOO radical. Unstructured bands observed in both spectral regions are assigned to cis-DOOO. Nascent OD X(2)Pi product state distributions following vibrational predissociation appear to be nearly statistical with respect to the degree of rotational excitation, but display a marked propensity for Pi(A') Lambda-doublets, which is interpreted as a signature of a planar dissociation. The energetically highest open OD X(2)Pi product channel implies an upper limit dissociation energy D0 < or = 1856 cm(-1) or 5.31 kcal mol(-1). This value allows refinement of the upper limit D0 of the atmospherically important HOOO isotopomer, suggesting that it is marginally less stable than previously thought. PMID- 18507451 TI - Probing the molecular and electronic structure of capsaicin: a spectroscopic and quantum mechanical study. AB - The conformational preferences of capsaicin were investigated by using the hybrid meta density functional theory (DFT) method MPWB1K. Its flexible, pendant side chain allows for a multitude of conformations only slightly different in energy. The distinctive vibrational features of the most stable conformers were characterized. To elucidate the most favorable reaction sites of capsaicin for radical scavenging, various homolytic bond-dissociation energies were also calculated. Of the possible radical intermediates, the allyl and benzyl radicals are energetically preferred. The filled and empty electronic structures of capsaicin were investigated by exploiting the photoelectron and electron transmission spectra also of reference molecules and suitable quantum-mechanical calculations. On this basis, a reliable pattern of the vertical ionization energies and electron-attachment energies of capsaicin was proposed. The frontier pi molecular orbitals are concentrated over the vanillyl moiety, with a modest influence of the amidic-aliphatic chain. The (negative) first vertical electron affinity is predicted to be similar to that of benzene. The absorption spectrum of capsaicin and its change by conversion into a phenolic deprotonated anion (modest bathochromic displacement) or a phenoxyl neutral radical (from colorless to red) were interpreted with time-dependent DFT calculations. ESR measurements following chemical or electrochemical reduction of capsaicin did not lead to detection of the corresponding radical anion. The spectra show fragmentation of the original molecule and formation of a variety of radical species which are believed to have a semiquinonic structure. PMID- 18507452 TI - Continuum polarizable force field within the Poisson-Boltzmann framework. AB - We have developed and tested a complete set of nonbonded parameters for a continuum polarizable force field. Our analysis shows that the new continuum polarizable model is consistent with B3LYP/cc-pVTZ in modeling electronic response upon variation of dielectric environment. Comparison with experiment also shows that the new continuum polarizable model is reasonable, with accuracy similar to that of B3LYP/cc-pVTZ in reproduction of dipole moments of selected organic molecules in the gas phase. We have further tested the validity to interchange the Amber van der Waals parameters between the explicit and continuum polarizable force fields with a series of dimers. It can be found that the continuum polarizable model agrees well with MP2/cc-pVTZ, with deviations in dimer binding energies less than 0.9 kcal/mol in the aqueous dielectric environment. Finally, we have optimized atomic cavity radii with respect to experimental solvation free energies of 177 training molecules. To validate the optimized cavity radii, we have tested these parameters against 176 test molecules. It is found that the optimized Poisson-Boltzmann atomic cavity radii transfer well from the training set to the test set, with an overall root-mean square deviation of 1.30 kcal/mol, an unsigned average error of 1.07 kcal/mol, and a correlation coefficient of 92% for all 353 molecules in both the training and test sets. Given the development documented here, the next natural step is the construction of a full protein/nucleic acid force field within the new continuum polarization framework. PMID- 18507453 TI - Interactions between spiropyrans and room-temperature ionic liquids: photochromism and solvatochromism. AB - A series of imidazolium-based room-temperature ionic liquids (RTILs) containing anions from organic carboxylic acids were prepared. A set of dye probes, including Reichardt's dye (30), 4-nitrioaniline, and N, N-diethyl-4-nitroaniline, were used to determine the ET(30) scales and the Kamlet-Taft parameters (pi*, alpha, and beta) of the RTILs. On the basis of the polarity properties, these RTILs were categorized into three groups: group A with beta >0.9, alpha <0.9; group B with beta <0.9, alpha <0.9; and group C with beta <0.9, alpha >0.9. Interactions of these RTILs with four photochromic spiropyran derivatives (SP-I, SP-II, SP-III, and SP-IV) were investigated. It was found that the spiropyrans could present photochromism (positive or negative) or not, depending mainly on the polarity properties of the RTILs and also on the structure itself. A new spectroscopic method based on the molecular transition energy of the spiropyran probes (ESP) was proposed to determine the polarity of those protic or fluorine containing RTILs, which were failed with the Reichardt's dye (30) probe. PMID- 18507454 TI - Extraction of electrode kinetic parameters from microdisc voltammetric data measured under transport conditions intermediate between steady-state convergent and transient linear diffusion as typically applies to room temperature ionic liquids. AB - The extraction of electrode kinetic parameters for electrochemical couples in room-temperature ionic liquids (RTILs) is currently an area of considerable interest. Electrochemists typically measure electrode kinetics in the limits of either transient planar or steady-state convergent diffusion for which the voltammetric response is well understood. In this paper we develop a general method allowing the extraction of this kinetic data in the region where the diffusion is intermediate between the planar and convergent limits, such as is often encountered in RTILs using microelectrode voltammetry. A general working surface is derived, allowing the inference of Butler-Volmer standard electrochemical rate constants for the peak-to-peak potential separation in a cyclic voltammogram as a function of voltage scan rate. The method is applied to the case of the ferrocene/ferrocenium couple in [C2mim][N(Tf)2] and [C4mim][N(Tf)2]. PMID- 18507455 TI - Hollow single-crystal spinel nanocubes: the case of zinc cobalt oxide grown by a unique Kirkendall effect. AB - Small hollow nanocubes of the ternary spinel Zn x Co 1- x Co 2O 4 of ca. 18 nm dimension were prepared via a facile hydrothermal route. A growth mechanism is suggested in which solid single-crystal Co 3O 4 nanocubes are gradually converted to hollow single-crystal Zn x Co 1- x Co 2O 4 nanocubes with preservation of the spinel framework through differential diffusion of Zn (2+) and Co (2+) ions. With the cation exchange, the chemical composition and thus physical properties can be tailored. PMID- 18507456 TI - Synthesis, structural characterization, and ligand replacement reactions of gem dithiolato-bridged rhodium and iridium complexes. AB - The reaction of gem-dithiol compounds R 2C(SH) 2 (R = Bn (benzyl), (i) Pr; R 2 = (CH 2) 4-) with dinuclear rhodium or iridium complexes containing basic ligands such as [M(mu-OH)(cod)] 2 and [M(mu-OMe)(cod)] 2, or the mononuclear [M(acac)(cod)] (M = Rh, Ir, cod = 1,5-cyclooctadiene) in the presence of a external base, afforded the dinuclear complexes [M 2(mu-S 2CR 2)(cod) 2] ( 1- 4). The monodeprotonation of 1,1-dimercaptocyclopentane gave the mononuclear complex [Rh(HS 2Cptn)(cod)] ( 5) that is a precursor for the dinuclear compound [Rh 2(mu S 2Cptn)(cod) 2] ( 6). Carbonylation of the diolefin compounds gave the complexes [Rh 2(mu-S 2CR 2)(CO) 4] ( 7- 9), which reacted with P-donor ligands to stereoselectively produce the trans isomer of the disubstituted complexes [Rh 2(mu-S 2CR 2)(CO) 2(PR' 3) 2] (R' = Ph, Cy (cyclohexyl)) ( 10- 13) and [Rh 2(mu-S 2CBn 2)(CO) 2{P(OR') 3} 2] (R' = Me, Ph) ( 14- 15). The substitution process in [Rh 2(mu-S 2CBn 2)(CO) 4] ( 7) by P(OMe) 3 has been studied by spectroscopic means and the full series of substituted complexes [Rh 2(mu-S 2CBn 2)(CO) 4- n {P(OR) 3} n ] ( n = 1, 4) has been identified in solution. The cis complex [Rh 2(mu-S 2CBn 2)(CO) 2(mu-dppb)] ( 16) was obtained by reaction of 7 with the diphosphine dppb (1,4-bis(diphenylphosphino)butane). The molecular structures of the diolefinic dinuclear complexes [Rh 2(mu-S 2CR 2)(cod) 2] (R = Bn ( 1), (i) Pr ( 2); R 2 = -(CH 2) 4- ( 6)) and that of the cis complex 16 have been studied by X-ray diffraction. PMID- 18507457 TI - Structural and spectroscopic impact of tuning the stereochemical activity of the lone pair in lead(II) cyanoaurate coordination polymers via ancillary ligands. AB - The reaction of Pb(ClO4)2 x xH2O, an ancillary ligand L, and two equivalents of Au(CN)2(-) gave a series of crystalline coordination polymers, which were structurally characterized. The ligands were chosen to represent a range of increasing basicity, to influence the stereochemical activity (i.e., p-orbital character) of the Pb(II) lone pair. The Pb(II) center in [Pb(1,10 phenanthroline)2][Au(CN)2]2 (1) is 8-coordinate, with a stereochemically inactive lone pair; all 8 Pb-N bonds are similar. The Au(CN)2(-) units propagate a 2-D brick-wall structure. In [Pb(2,2'-bipyridine)2][Au(CN)2]2 (2), the 8-coordinate Pb(II) center has asymmetric Pb-N bond lengths, indicating moderate lone pair stereochemical activity; the supramolecular structure forms a 1-D chain/ribbon motif. For [Pb(ethylenediamine)][Au(CN)2]2 (3), the Pb(II) is only 5-coordinate and extremely asymmetric, with Pb-N bond lengths from 2.123(7) to 3.035(9) A; a rare Pb-Au contact of 3.5494(5) A is also observed. The Au(CN)2(-) units connect the Pb(ethylenediamine) centers to form 1-D zigzag chains which stack via Au-Au interactions of 3.3221(5) A to yield a 2-D sheet. (207)Pb MAS NMR of the polymers indicates an increase in both the chemical shielding span and isotropic chemical shift with increasing Pb(II) coordination sphere anisotropy (from delta iso = 2970 and Omega = 740 for 1 to delta iso = -448 and Omega = 3980 for 3). The shielding anisotropy is positively correlated with Pb(II) p-character, and reflects a direct connection between the NMR parameters and lone-pair activity. Solid-state variable-temperature luminescence measurements indicate that the emission bands at 520 and 494 nm, for 1 and 2, respectively, can be attributed to Pb --> L transitions, by comparison with simple [Pb(L)2](ClO4)2 salts. In contrast, two emission bands for 3 at 408 and 440 nm are assignable to Au-Au and Pb-Au-based transitions, respectively, as supported by single-point density functional theory calculations on models of 3. PMID- 18507458 TI - Au dimers on thin MgO(001) films: flat and charged or upright and neutral? AB - A combination of low temperature scanning tunneling microscopy (STM) and theoretical calculations is used to investigate Au dimers, supported on thin MgO(001) films, whose thickness was chosen such that charge transfer from the Ag substrate to the deposited Au is possible. Au dimers exist not only in an upright geometry--as theoretically predicted to be the most stable configuration--but also as flat lying dimers which populate a manifold of different azimuthal orientations. Apart from the difference in adsorption configurations, these two isomers exhibit rather different electronic structures: while upright dimers are neutral, flat ones are charged. PMID- 18507459 TI - Reaction of carboxylic acids and isonitriles in small spaces. AB - Reversible encapsulation complexes are applied to the reaction of isonitriles with carboxylic acids. Encapsulation facilitates these reactions by amplifying the concentration of reactants, arranging the acid and isonitrile in the appropriate orientations, isolating intermediates from bulk solution, and providing an organized solvent cage. PMID- 18507460 TI - Conformationally driven asymmetric induction of a catalytic dendrimer. AB - Nature's catalysts promote the reactions necessary for life with extremely high specificity by folding into specific shapes capable of communicating remote structural information to an active site. Achieving this objective in synthetic systems has been hampered by the lack of information concerning how dynamic conformational chirality can influence the stereoselectivity of a catalytic process. Herein, we report the first illustration of a catalytic dendrimer that achieves high enantioselectivity by amplifying/propagating local chirality via a dynamically folded structure. Experimental evidence supports a chiral relay mechanism that propagates local terminal chirality of the dendron to the axial chirality of the biphenyl core through the helical secondary structure of the dendron. PMID- 18507461 TI - Diacid linkers that promote parallel beta-sheet secondary structure in water. AB - We report the development of diacid units that promote formation of a two stranded parallel beta-sheet secondary structure between peptide segments attached via their N-termini. These linker units are formed by attaching glycine to one carboxyl group of cis-1,2-cyclohexanedicarboxylic acid (CHDA). Parallel sheet formation in water is observed when l-residue strands are attached to the CHDA-Gly unit with either of the two absolute configurations. PMID- 18507462 TI - Molecular recognition based on low-affinity polyvalent interactions: selective binding of a carboxylated polymer to fibronectin fibrils of live fibroblast cells. AB - To explore molecular recognition of biomolecules in the complex environment of the extracellular matrix, we utilized two fluorescent poly(p phenyleneethynylene)s bearing either cationic alkylammonium or negatively charged carboyxlate side chains. While incubation of live NIH 3T3 fibroblast cells with the cationic polymer yielded perinuclear punctate staining reminiscent of endocytotic vesicles, the carboxylated polymer revealed a characteristic filamentous staining pattern. Histochemical and immunofluorescence studies demonstrated that the anionic PPE selectively binds to fibronectin fibrils of the extracellular matrix. An in vitro binding study revealed a dissociation constant of approximately 100 nM for the fibronectin-polymer complex. Both polymers showed bright two-photon excited emission as well as low toxicity, rendering them well suited for live cell imaging studies. The studies demonstrate that selective molecular recognition of biomolecules in the complex environment of the extracellular matrix can be achieved by means of nonspecific low-affinity polyvalent interactions. PMID- 18507463 TI - Synthesis of cadmium telluride quantum wires and the similarity of their effective band gaps to those of equidiameter cadmium telluride quantum dots. AB - High-quality colloidal CdTe quantum wires having purposefully controlled diameters in the range 5-11 nm are grown by the solution-liquid-solid (SLS) method, using Bi nanoparticle catalysts, cadmium octadecylphosphonate and trioctylphosphine telluride as precursors, and a TOPO solvent. The wires adopt the wurtzite structure and grow along the [002] direction (parallel to the c axis). The size dependence of the effective band gaps in the wires is determined from the absorption spectra and compared to the experimental results for high quality CdTe quantum dots. In contrast to the predictions of an effective-mass approximation, particle-in-a-box model, and previous experimental results from CdSe and InP dot-wire comparisons, the effective band gaps of CdTe dots and wires of like diameter are found to be experimentally indistinguishable. The present results are analyzed using density functional theory under the local-density approximation by implementing a charge-patching method. The higher-level theoretical analysis finds the general existence of a threshold diameter, above which dot and wire effective band gaps converge. The origin and magnitude of this threshold diameter are discussed. PMID- 18507464 TI - Growth, new growth, and amplification of carbon nanotubes as a function of catalyst composition. AB - Carbon nanotubes (CNTs) have been grown using Fe, Co, Ni, and Co/Fe spin-on catalyst (SOC) systems, involving the metal salt dispersed with a spin-on-glass precursor. During initial growth runs (CH4/H2/900 degrees C), the CNT yield followed the order Co-SOC > Fe-SOC >> Ni-SOC. The Fe catalysts produced the longest nanotubes at the expense of a larger average CNT diameter and broader diameter distribution than the Co-SOC system. A series of Co/Fe-SOCs were prepared where as the atomic percentage of Co is increased nucleation of CNT increases but the CNT length decreases. The linear relationship between the diameter and length of CNTs grown from the Co/Fe-SOC suggests that slow growth is beneficial with respect to control over CNT diameter. After initial CNT growth, the original samples were subjected to additional growth runs. Four individual reactions were observed in the Fe-SOC and binary Co/Fe-SOC: regrowth (amplification), double growth (a second CNT growing from a previously active catalyst), CNT etching, and nucleation from initially inactive catalysts (new growth). CNT etching was observed for the mixed catalyst systems (Co/Fe-SOC) but not for either Fe-SOC or Co-SOC. During the regrowth experiments, CNTs were observed that were not present after the initial growth run (and were not as a result of amplification or double growth). Thus, catalysts, which were initially inactive toward nucleation of CNTs in the original growth run, are capable of becoming activated when placed back into the furnace and submitted to regrowth under identical conditions. PMID- 18507465 TI - Comparative structural and chemical studies of ferritin cores with gradual removal of their iron contents. AB - Transmission Electron Microscopy (TEM), X-ray Absorption Near Edge Spectroscopy (XANES), Electron Energy-Loss Spectroscopy (EELS), Small-Angle X-ray Scattering (SAXS), and SQUID magnetic studies were performed in a batch of horse spleen ferritins from which iron had been gradually removed, yielding samples containing 2200, 1200, 500, and 200 iron atoms. Taken together, findings obtained demonstrate that the ferritin iron core consists of a polyphasic structure (ferrihydrite, magnetite, hematite) and that the proportion of phases is modified by iron removal. Thus, the relative amount of magnetite in ferritin containing 2200 to 200 iron atoms rose steadily from approximately 20% to approximately 70% whereas the percentage of ferrihydrite fell from approximately 60% to approximately 20%. These results indicate a ferrihydrite-magnetite core-shell structure. It was also found that the magnetite in the ferritin iron core is not a source of free toxic ferrous iron, as previously believed. Therefore, the presence of magnetite in the ferritin cores of patients with Alzheimer's disease is not a cause of their increased brain iron(II) concentration. PMID- 18507466 TI - Pyrolysis, crystallization, and sintering of mesostructured titania thin films assessed by in situ thermal ellipsometry. AB - In-situ thermal ellipsometric analysis is used to elucidate new and fine-scale details on the thermally driven densification, pyrolysis, crystallization, and sintering of dense and ordered mesoporous titania thin films prepared by evaporation-induced self-assembly. The role of the heating schedule, initial film thickness, nature of the substrate and templating agent, solution aging, and presence of water and other additives in the calcination environment is examined. Each of these parameters is shown to have unique and often substantial effects on the final film structure, while the technique itself provides detailed insight into the chemical origin and evolution of these effects. In-situ monitoring and control over the governing chemical processes, such as high-temperature adsorption phenomena that impact nanocrystal growth, is also demonstrated. The evolution of both the porosity and chemical processes occurring inside these materials are evaluated, including extraction of kinetic parameters for the pyrolysis of the template and crystallization of the matrix walls. The latter is shown to be strongly dependent on the presence of mesoscale ordering with ordered cubic films indicating a 1D diffusion-limited crystallization process and dense films following a 3D diffusion-limited process. Less well-ordered mesoporous films, despite similarities in pore volume and pore size distributions, are kinetically more reminiscent of dense films in terms of crystallization. In-situ thermal ellipsometry, by detailing the evolution of the thermally driven chemistry and ceramization that dictate the final film properties, provides immensely important insight into the synthesis and optimization of advanced functional materials based on titania and other metal oxide thin films. PMID- 18507467 TI - Electronically activated actin protein polymerization and alignment. AB - Biological systems are the paragon of dynamic self-assembly, using a combination of spatially localized protein complexation, ion concentration, and protein modification to coordinate a diverse set of self-assembling components. Biomimetic materials based upon biologically inspired design principles or biological components have had some success at replicating these traits, but have difficulty capturing the dynamic aspects and diversity of biological self assembly. Here, we demonstrate that the polymerization of ion-sensitive proteins can be dynamically regulated using electronically enhanced ion mixing and monomer concentration. Initially, the global activity of the cytoskeletal protein actin is inhibited using a low-ionic strength buffer that minimizes ion complexation and protein-protein interactions. Nucleation and growth of actin filaments are then triggered by a low-frequency AC voltage, which causes local enhancement of the actin monomer concentration and mixing with Mg(2+). The location and extent of polymerization are governed by the voltage and frequency, producing highly ordered structures unprecedented in bulk experiments. Polymerization rate and filament orientation could be independently controlled using a combination of low frequency (approximately 100 Hz) and high frequency (1 MHz) AC voltages, creating a range of macromolecular architectures from network hydrogel microparticles to highly aligned arrays of actin filaments with approximately 750 nm periodicity. Since a wide range of proteins are activated upon complexation with charged species, this approach may be generally applicable to a variety of biopolymers and proteins. PMID- 18507468 TI - Regio- and chemoselective multiple functionalization of pyrimidine derivatives by selective magnesiations using TMPMgCl.LiCl. AB - Successive regio- and chemoselective magnesiations of pyrimidines using TMPMgCl.LiCl furnish, after trapping with various electrophiles, highly functionalized derivatives in good to excellent yields. Applications to the synthesis of antiviral and anti-inflammatory agents such as p38 and sPLA2 kinase inhibitors are reported. PMID- 18507469 TI - New cationic liposomes as vehicles of m-tetrahydroxyphenylchlorin in photodynamic therapy of infectious diseases. AB - Antimicrobial photodynamic therapy is emerging as a promising therapeutic modality for bacterial infections. For optimizing the antibacterial activity of the photosensitizer m-tetrahydroxyphenylchlorin, it has been encapsulated in mixed cationic liposomes composed of different ratios of dimyristoyl- sn-glycero phosphatidylcholine and any of four cationic surfactants derived from l-prolinol. The delivery efficiency of the different liposomes formulations has been evaluated on a methicillin-resistant Staphylococcus aureus bacterial strain (MRSA), and one of the tested formulations shows a biological activity comparable to that of the free chlorin. In order to rationalize the physicochemical parameters of the carriers that control the biological activity, the new liposome formulations have been characterized by measuring (a) the zeta potential, (b) their capability of chlorin entrapping efficiency, i.e. entrapment efficacy, (c) the effect of storage on chlorin entrapment and (d) the localization of chlorin in the bilayer. The correlation of the physicochemical and biological features of formulations has allowed us to rationalize, to some extent, some of the parameters that may control the interactions with the biological environment. PMID- 18507470 TI - Synthesis of bisquinolone-based mono- and diphosphine ligands of the Aza-BINAP type. AB - Mono- and bisphosphine ligands based on the 4,4'-bisquinolone structural framework (BIQUIP ligands) were generated by direct microwave-assisted palladium catalyzed carbon-phosphorus cross-coupling reactions employing the corresponding heteroaryl bromides and diphenylphosphine as substrates. PMID- 18507471 TI - Flexusines A and B and epimukulol from the soft coral Sarcophyton flexuosum. AB - Three new cembranes, flexusines A (1) and B (2) and an epimukulol (3), were isolated from the soft coral Sarcophyton flexuosum collected near Reunion Island, Indian Ocean. Their structures were elucidated using 1D and 2D NMR spectral analyses. PMID- 18507472 TI - Cytotoxic dihydroagarofuranoid sesquiterpenes from the seeds of Celastrus orbiculatus. AB - A chemical study on the seeds of Celastrus orbiculatus has led to the isolation of nine new (1- 9) and 13 known dihydro-beta-agarofuran derivatives. The identification and structural elucidation of the new compounds were based on spectroscopic data analysis, and the absolute configurations of compounds 1- 6, 8 10, and 16, as well as derivatives 2a and 6a, were determined by CD studies or by chemical methods. All compounds isolated were evaluated for cytotoxic activity against HL-60 human leukemia cells. PMID- 18507473 TI - Isolation, structure elucidation, and synthesis of cytotoxic tryptanthrin analogues from Phaius mishmensis. AB - Bioassay-guided chromatographic separation of the cytotoxic MeOH extract of Phaius mishmensis led to the isolation of two known and six new indoloquinazolinones, phaitanthrins A-E (1-5) and methylisatoid (6). The structures of the new compounds were elucidated by spectroscopic analysis. Phaitanthrin A (1) and tryptanthrin (7) showed moderate cytotoxicity against MCF 7, NCI-H460, and SF-268 cell lines. A series of ketone adducts of tryptanthrin were prepared and tested initially for anticancer activity in vitro against MCF 7, NCI-H460, and SF-268 human cancer cell lines. The 3-pentanone adduct 13 showed activity similar to tryptanthrin. PMID- 18507474 TI - Novel open-chain cytochalsins from the marine-derived fungus Spicaria elegans. AB - Six novel open-chain cytochalasins (1-6) and one known [12]-cytochalasin (7) have been isolated from the fermentation broth of a marine-derived fungus, Spicaria elegans. Cytochalasins Z10-Z15 (1-6) are the first reported cytochalasins that contain an open chain to date. The structures of these new compounds were elucidated by spectroscopic methods. The cytotoxic effects on P388, A-549, HL-60, and BEL-7402 cell lines of all compounds were evaluated by the MTT method. PMID- 18507475 TI - High-throughput synthesis of HepDirect prodrugs of nucleoside monophosphates. AB - A high-throughput phosphoramidite method for HepDirect prodrug synthesis was optimized on seven representative nucleosides, adenosine, inosine, guanosine, uridine, cytidine, AICA-riboside, and thymidine, each on a 5 mg scale. The variables optimized included (1) reaction time, (2) reaction temperature, (3) activating agent, (4) solvent, (5) purification method, and (6) stoichiometry. Preparative HPLC with mass-based fraction collection and yield determination from an ELSD standard curve enabled high-throughput. The optimized conditions for the representative nucleosides required 6 mol equiv of phosphoramidite to nucleoside and resulted in an average HPLC determined yield of 31 +/- 14% and HPLC purity of 93 +/- 3%. PMID- 18507476 TI - Synthesis of substituted benzofurans via microwave-enhanced catch and release strategy. AB - A microwave-enhanced procedure for the synthesis of substituted benzofurans starting from 2-(1-hydroxyalkyl)-phenols and using triphenylphosphine polystirene resin is reported. The benzofurans are isolated in good to high yields and purities by simple workup. The procedure can be applied to chiral alpha-alkyl-2 benzofuranmethanamines too. PMID- 18507477 TI - Reducing thermal conductivity of crystalline solids at high temperature using embedded nanostructures. AB - Thermal conductivity of a crystalline solid at high temperature is dominated by the Umklapp process because the number of high frequency phonons increases with temperature. It is challenging to reduce the thermal conductivity of crystalline solids at high temperature although it is widely known that, by increasing the atomic defect concentration, thermal conductivity of crystalline solids can be reduced at low temperature. By increasing the concentration of ErAs nanoparticles in In 0.53Ga 0.47As up to 6 atom %, we demonstrate a thermal conductivity reduction by almost a factor of 3 below that of In 0.53Ga 0.47As at high temperature. A theoretical model suggests that the mean free path of the low frequency phonons is suppressed by increasing the ErAs nanoparticle concentration. PMID- 18507478 TI - Imaging dielectric properties of Si nanowire oxide with conductive atomic force microscopy complemented with femtosecond laser illumination. AB - In most Si nanowire (NW) applications, Si oxide provides insulation or a medium of controlled electron tunneling. This work revealed both similarities and differences in the dielectric properties of NW oxide compared with that grown on wafers. The interface barrier to electron transit from the semiconductor to the dielectric and the threshold electric field for current flow are quite similar to those in the planar geometry. This is not true for the lowest currents measured which are not uniformly distributed, indicating variations of trap density in the gap of NW oxide. PMID- 18507479 TI - Strong antimicrobial coatings: single-walled carbon nanotubes armored with biopolymers. AB - Large scale biomimetic single-walled carbon nanotube (SWNT) coatings with significant antimicrobial activity, high Young's Modulus, and controlled morphology were fabricated using layer-by-layer assembly. Thickness was controlled within 1.6 nm and SWNT orientation was controlled using a directed air stream. This unique blend of multifunctionality and vertical and lateral control of a bottom-up assembly process is a significant advancement in developing macroscale assemblies with the combined attributes of SWNTs and natural materials. PMID- 18507480 TI - Controlled manipulation of giant hybrid inorganic nanowire assemblies. AB - The ultimate goal of nanotechnology is the design and fabrication of nanosize building blocks with multiple functionalities and their assembly into large-scale functional structures that can be controllably manipulated. Here we show that hybrid inorganic multisegmented nanowires, with hydrophobic carbon nanotube tails and hydrophilic metal nanowire heads, allow the assembly and manipulation of massive ordered structures in solution, reminiscent of the organic molecular micellar assembly. Further, properly designed assemblies can be manipulated using external stimuli such as magnetic field and light. The hybrid nanowires can have multiple segments including magnetic components, allowing the assembly to be manipulated by external magnetic field. The assembled structures can also be manipulated by modifying the hydrophobicity of the respective components via chemical functionalization and optical irradiation. This approach brings the concept of environment sensitive self-assembling nanomaterials closer to reality. PMID- 18507481 TI - On the polyol synthesis of silver nanostructures: glycolaldehyde as a reducing agent. AB - The polyol synthesis is a popular method of preparing metal nanostructures, yet the mechanism by which metal ions are reduced is poorly understood. Using a spectrophotometric method, we show, for the first time, that heating ethylene glycol (EG) in air results in its oxidation to glycolaldehyde (GA), a reductant capable of reducing most noble metal ions. The dependence of reducing power on temperature for EG can be explained by this temperature-dependent oxidation, and the factors influencing GA production can have a profound impact on the nucleation and growth kinetics. These new findings provide critical insight into how the polyol synthesis can be used to generate metal nanostructures with well controlled shapes. For example, with the primary reductant identified, it becomes possible to evaluate and understand its explicit role in generating nanostructures of a specific shape to the exclusion of others. PMID- 18507482 TI - Synthesis of nanometer-sized sodalite without adding organic additives. AB - Aggregates (80 nm) of sodalite nanocrystals with crystallite sizes ranging from 20 to 40 nm have been synthesized from a sodium aluminosilicate solution at low temperature, without adding any organic additives, while paying attention to the key factors for the synthesis of nanosized zeolite crystals. The physical properties of nanosized sodalite crystals were characterized by X-ray diffraction, scanning electron microscopy, high-resolution transmission electron microscopy, 29Si solid-state magic-angle spinning (MAS) NMR, and N2 adsorption. As expected, the external surface area of nanosized sodalite crystals is significantly increased compared with that of microsized sodalite crystals. The size of synthesized sodalite crystals can be controlled from 20 nm to 10 microm. It is found that the preparation of a homogeneous aluminosilicate solution followed by the formation of an aluminosilicate hard gel by adjusting the initial composition, for example, SiO2/Al2O3 and Na2O/H2O ratios, is critical for synthesis. PMID- 18507483 TI - Acute treatment of patients with bipolar mixed episodes. AB - Managing bipolar mixed states should start by establishing a diagnosis, assessing for comorbidities, and clarifying targets for therapy. Differentiating between unipolar and bipolar disorder is important because these conditions have dissimilar efficacious pharmacotherapeutic options. To treat mixed states, atypical antipsychotics or divalproex have been shown to be effective, and antidepressants should generally be avoided. Because of the complexity of mixed episodes, monotherapy may be insufficient and patients may require combination treatment. As more medications become available in the treatment of bipolar disorder, algorithms can help clinicians make the appropriate treatment choices by offering recommendations for sequencing agents. PMID- 18507484 TI - Relapse prevention in patients with schizophrenia. AB - Remission is a realistic goal for patients with schizophrenia, and, if sustained remission can be achieved, then patients may eventually attain functional recovery. The chance of relapse is decreased if patients are adherent to their pharmacotherapeutic treatment regimen. This case explores how to prevent relapse for a patient with schizophrenia and comorbid substance use disorder who is nonadherent to an oral atypical antipsychotic. To help the patient achieve treatment goals, intervention strategies such as switching medications and implementing psychosocial therapies are considered. PMID- 18507485 TI - Defining the bipolar spectrum and treating bipolar II disorder. AB - The treatment of bipolar II disorder may be complicated by the lack of a universal definition of the bipolar spectrum and by the limited number of studies focusing on bipolar II disorder pharmacotherapy. The appropriate first-line treatment for bipolar II disorder is still being studied, but according to the limited research, antidepressants, mood stabilizers, anticonvulsants, and atypical antipsychotics have been safe and effective in the acute and maintenance treatment of bipolar II depression and/or hypomania or mania. A consensus should be reached on the definition of the bipolar spectrum, and further research is needed to determine the best first-line treatment for bipolar II disorder. PMID- 18507486 TI - Perinatal psychiatry: risk factors, treatment data, and specific challenges for clinical researchers. PMID- 18507487 TI - Antipsychotic polypharmacy, part 1: Shotgun approach or targeted cotreatment? PMID- 18507488 TI - Late-onset hypersensitivity reaction with leukopenia and thrombocytopenia induced by oxcarbazepine treatment in a patient with schizoaffective disorder. PMID- 18507489 TI - Reasons for inadequate utilization of cognitive-behavioral therapy for obsessive compulsive disorder. PMID- 18507490 TI - Metabolic syndrome in patients with bipolar disorder. PMID- 18507491 TI - Effect of ribavirin, in combination with interferon in patients with hepatitis C, on the bleeding risk associated with selective serotonin reuptake inhibitors. PMID- 18507493 TI - Comments on a randomized, double-blind comparison of sertraline and placebo for posttraumatic stress disorder in a Department of Veterans Affairs setting. PMID- 18507494 TI - Increasing rates of placebo response over time in mania studies. PMID- 18507496 TI - How do US journalists cover treatments, tests, products, and procedures? An evaluation of 500 stories. PMID- 18507497 TI - Association of prenatal and childhood blood lead concentrations with criminal arrests in early adulthood. AB - BACKGROUND: Childhood lead exposure is a purported risk factor for antisocial behavior, but prior studies either relied on indirect measures of exposure or did not follow participants into adulthood to examine the relationship between lead exposure and criminal activity in young adults. The objective of this study was to determine if prenatal and childhood blood lead concentrations are associated with arrests for criminal offenses. METHODS AND FINDINGS: Pregnant women were recruited from four prenatal clinics in Cincinnati, Ohio if they resided in areas of the city with a high concentration of older, lead-contaminated housing. We studied 250 individuals, 19 to 24 y of age, out of 376 children who were recruited at birth between 1979 and 1984. Prenatal maternal blood lead concentrations were measured during the first or early second trimester of pregnancy. Childhood blood lead concentrations were measured on a quarterly and biannual basis through 6.5 y. Study participants were examined at an inner-city pediatric clinic and the Cincinnati Children's Hospital Medical Center in Cincinnati, Ohio. Total arrests and arrests for offenses involving violence were collected from official Hamilton County, Ohio criminal justice records. Main outcomes were the covariate-adjusted rate ratios (RR) for total arrests and arrests for violent crimes associated with each 5 microg/dl (0.24 micromol/l) increase in blood lead concentration. Adjusted total arrest rates were greater for each 5 microg/dl (0.24 micromol/l) increase in blood lead concentration: RR = 1.40 (95% confidence interval [CI] 1.07-1.85) for prenatal blood lead, 1.07 (95% CI 0.88-1.29) for average childhood blood lead, and 1.27 (95% CI 1.03-1.57) for 6 year blood lead. Adjusted arrest rates for violent crimes were also greater for each 5 microg/dl increase in blood lead: RR = 1.34 (95% CI 0.88-2.03) for prenatal blood lead, 1.30 (95% CI 1.03-1.64) for average childhood blood lead, and 1.48 (95% CI 1.15-1.89) for 6-year blood lead. CONCLUSIONS: Prenatal and postnatal blood lead concentrations are associated with higher rates of total arrests and/or arrests for offenses involving violence. This is the first prospective study to demonstrate an association between developmental exposure to lead and adult criminal behavior. PMID- 18507498 TI - Disease mongering is now part of the global health debate. PMID- 18507499 TI - Decreased brain volume in adults with childhood lead exposure. AB - BACKGROUND: Although environmental lead exposure is associated with significant deficits in cognition, executive functions, social behaviors, and motor abilities, the neuroanatomical basis for these impairments remains poorly understood. In this study, we examined the relationship between childhood lead exposure and adult brain volume using magnetic resonance imaging (MRI). We also explored how volume changes correlate with historic neuropsychological assessments. METHODS AND FINDINGS: Volumetric analyses of whole brain MRI data revealed significant decreases in brain volume associated with childhood blood lead concentrations. Using conservative, minimum contiguous cluster size and statistical criteria (700 voxels, unadjusted p < 0.001), approximately 1.2% of the total gray matter was significantly and inversely associated with mean childhood blood lead concentration. The most affected regions included frontal gray matter, specifically the anterior cingulate cortex (ACC). Areas of lead associated gray matter volume loss were much larger and more significant in men than women. We found that fine motor factor scores positively correlated with gray matter volume in the cerebellar hemispheres; adding blood lead concentrations as a variable to the model attenuated this correlation. CONCLUSIONS: Childhood lead exposure is associated with region-specific reductions in adult gray matter volume. Affected regions include the portions of the prefrontal cortex and ACC responsible for executive functions, mood regulation, and decision-making. These neuroanatomical findings were more pronounced for males, suggesting that lead-related atrophic changes have a disparate impact across sexes. This analysis suggests that adverse cognitive and behavioral outcomes may be related to lead's effect on brain development producing persistent alterations in structure. Using a simple model, we found that blood lead concentration mediates brain volume and fine motor function. PMID- 18507501 TI - Neurological and behavioral consequences of childhood lead exposure. PMID- 18507500 TI - Convergence of mutation and epigenetic alterations identifies common genes in cancer that predict for poor prognosis. AB - BACKGROUND: The identification and characterization of tumor suppressor genes has enhanced our understanding of the biology of cancer and enabled the development of new diagnostic and therapeutic modalities. Whereas in past decades, a handful of tumor suppressors have been slowly identified using techniques such as linkage analysis, large-scale sequencing of the cancer genome has enabled the rapid identification of a large number of genes that are mutated in cancer. However, determining which of these many genes play key roles in cancer development has proven challenging. Specifically, recent sequencing of human breast and colon cancers has revealed a large number of somatic gene mutations, but virtually all are heterozygous, occur at low frequency, and are tumor-type specific. We hypothesize that key tumor suppressor genes in cancer may be subject to mutation or hypermethylation. METHODS AND FINDINGS: Here, we show that combined genetic and epigenetic analysis of these genes reveals many with a higher putative tumor suppressor status than would otherwise be appreciated. At least 36 of the 189 genes newly recognized to be mutated are targets of promoter CpG island hypermethylation, often in both colon and breast cancer cell lines. Analyses of primary tumors show that 18 of these genes are hypermethylated strictly in primary cancers and often with an incidence that is much higher than for the mutations and which is not restricted to a single tumor-type. In the identical breast cancer cell lines in which the mutations were identified, hypermethylation is usually, but not always, mutually exclusive from genetic changes for a given tumor, and there is a high incidence of concomitant loss of expression. Sixteen out of 18 (89%) of these genes map to loci deleted in human cancers. Lastly, and most importantly, the reduced expression of a subset of these genes strongly correlates with poor clinical outcome. CONCLUSIONS: Using an unbiased genome-wide approach, our analysis has enabled the discovery of a number of clinically significant genes targeted by multiple modes of inactivation in breast and colon cancer. Importantly, we demonstrate that a subset of these genes predict strongly for poor clinical outcome. Our data define a set of genes that are targeted by both genetic and epigenetic events, predict for clinical prognosis, and are likely fundamentally important for cancer initiation or progression. PMID- 18507502 TI - False hopes, unwarranted fears: the trouble with medical news stories. PMID- 18507503 TI - Notch-deficient skin induces a lethal systemic B-lymphoproliferative disorder by secreting TSLP, a sentinel for epidermal integrity. AB - Epidermal keratinocytes form a highly organized stratified epithelium and sustain a competent barrier function together with dermal and hematopoietic cells. The Notch signaling pathway is a critical regulator of epidermal integrity. Here, we show that keratinocyte-specific deletion of total Notch signaling triggered a severe systemic B-lymphoproliferative disorder, causing death. RBP-j is the DNA binding partner of Notch, but both RBP-j-dependent and independent Notch signaling were necessary for proper epidermal differentiation and lipid deposition. Loss of both pathways caused a persistent defect in skin differentiation/barrier formation. In response, high levels of thymic stromal lymphopoietin (TSLP) were released into systemic circulation by Notch-deficient keratinocytes that failed to differentiate, starting in utero. Exposure to high TSLP levels during neonatal hematopoiesis resulted in drastic expansion of peripheral pre- and immature B-lymphocytes, causing B-lymphoproliferative disorder associated with major organ infiltration and subsequent death, a previously unappreciated systemic effect of TSLP. These observations demonstrate that local skin perturbations can drive a lethal systemic disease and have important implications for a wide range of humoral and autoimmune diseases with skin manifestations. PMID- 18507505 TI - Man versus machine versus ribozyme. PMID- 18507504 TI - Sperm sociality: cooperation, altruism, and spite. PMID- 18507507 TI - The promise of specialty pharmaceuticals: are they worth the price? AB - BACKGROUND: Specialty pharmaceuticals have evolved beyond their status as niche drugs designed to treat rare conditions and are now poised to become the standard of care in a wide variety of common chronic illnesses. Due in part to the cost of these therapies, payers are increasingly demanding evidence of their value. Determining the value of these medications is hampered by a lack of robust pharmacoeconomic data. OBJECTIVE: To outline emerging strategies and case study examples for the medical and pharmacy benefits management of specialty pharmaceuticals. SUMMARY: The promise of specialty pharmaceuticals: increased life expectancy, improved quality of life, enhanced workplace productivity, decreased burden of disease, and reduced health care spending comes at a significant cost. These agents require special handling, administration, patient education, clinical support, and risk mitigation. Additionally, specialty drugs require distribution systems that ensure appropriate patient selection and data collection. With the specialty pharmaceutical pipeline overflowing with new medicines and an aging population increasingly relying on these novel treatments to treat common diseases, the challenge of managing the costs associated with these agents can be daunting. Aided by sophisticated pharmacoeconomic models to assess value, the cost impacts of these specialty drugs can be appropriately controlled. CONCLUSION: Current evidence suggests that when used in targeted patient populations, specialty pharmaceuticals may represent a good health care value. PMID- 18507508 TI - Managing the intersection of medical and pharmacy benefits. AB - BACKGROUND: Because of the unique features of specialty pharmaceuticals and insurance plans, specialty pharmaceutical products can be paid through the pharmacy benefit or the medical benefit. While most pharmacists are very comfortable with the conventions of reimbursement in the pharmacy benefit, they are less familiar with the processes for payment in the medical benefit. OBJECTIVE: To review the 2 parallel processes for payment of specialty pharmaceuticals, the pharmacy benefit and the medical benefit, and to compare and contrast these 2 processes. SUMMARY: The medical benefit and pharmacy benefit processes for payment of specialty pharmaceuticals use different claim forms, product coding systems, pricing conventions, and contracts. Even though the services delivered can be identical, the financial aspects of paying for these services are quite different. CONCLUSION: Pharmacists who are interested in entering the specialty pharmacy arena, either as a provider or manager of providers, need to understand the payment processes for specialty pharmaceuticals through both the pharmacy and medical benefits. PMID- 18507509 TI - Benefit design innovations to manage specialty pharmaceuticals. AB - BACKGROUND: Spending on specialty pharmaceuticals is rising faster than that for traditional drugs. In 2006, specialty drugs were the largest category driving drug costs and utilization trends. Even with effective management, expenditures on these agents are projected to increase exponentially in the coming years. OBJECTIVE: To review benefit design strategies used by payers to control costs and manage utilization of specialty pharmaceuticals. SUMMARY: The rapid growth in specialty pharmaceutical expenditures reflects the introduction of new agents, broader indications, and wider use in more common disease states. The true growth of the specialty pharmaceutical segment is obscured because many of these agents are reimbursed through the medical benefit, which often lacks the transparency necessary to accurately determine true cost and utilization trends. To date, efforts to control spending on biologics have been fragmented with most payers employing techniques for cost and utilization containment similar to those used for traditional pharmaceuticals. To ensure greater cost and utilization control, a benefit design that simultaneously provides optimal cost management, appropriate utilization, improved clinical management, enhanced clinical outcomes, and heightened patient safety should be established. CONCLUSION: Current management techniques for specialty pharmaceuticals often represent a stop-gap approach for controlling rising drug costs. Creation of a specialty pharmacy benefit can optimize cost and utilization management. PMID- 18507510 TI - Health plan approach to operationalizing a specialty drug management program. AB - BACKGROUND: Expenditures related to specialty drugs consume a significant percentage of available health care resources. OBJECTIVE: Explain the process of transitioning the management of specialty drugs from medical services to pharmacy services in 2 managed care plans and provide insight into the issues encountered and solutions implemented based on 6 years of experience using traditional and innovative pharmacy utilization management tools to insure appropriate specialty drug use and reimbursement. SUMMARY: The level of involvement in a specialty management program varies from managing only products dispensed by the retail, mail, and specialty pharmacy vendor to encompassing a broad list of specialty drugs distributed through a variety of channels. Efficient administrative, operational, and clinical processes are critical to the success of the program. Additionally, an accurate and timely claims processing procedure is also essential for success as is the ability to mine data and effectively report on the use of specialty products. A clinically sound, cost-effective, and patient friendly program requires input from health plan members, pharmacy service leaders, and physician providers, and must overcome challenges associated with disrupting current relationships and removing competing incentives. CONCLUSION: A well-constructed and properly funded specialty drug management program results in clinical and financial benefits for the plan. PMID- 18507511 TI - Good news on the clinical gene transfer front. PMID- 18507512 TI - Good clinical practice standards: what they are and some tools to support them. PMID- 18507513 TI - Good clinical practice in the European Union. PMID- 18507514 TI - Suppression of renal cell carcinoma growth and metastasis with sustained antiangiogenic gene therapy. AB - Renal cell carcinoma (RCC) is the third most common urologic neoplasm. This aggressive malignancy has proven refractory to conventional treatment options. Antiangiogenic agents have shown early success in treating metastatic disease. The highly vascular nature of RCC appears particularly susceptible to this approach. This study investigates the potential of sustained expression of an endostatin-angiostatin fusion protein in an early-stage model of RCC to inhibit tumor growth and metastasis. Subcutaneous RCC-29 tumors were induced in athymic nude mice. Once tumors reached volumes of 10 and 25 mm(3), subjects received intratumoral injections of a nonreplicating adenoviral vector every 20 days until the conclusion of the trial. The mice were randomly assigned to three treatment groups: saline control, viral Ad-GFP control, and Ad-EndoAngio. Tumor volumes were measured twice weekly for 80 days. During days 40-50 of the trial, subjects underwent dual-photon optical imaging of the tumor vasculature to ascertain angiogenic changes. All animals underwent postmortem histopathological analysis to assess for metastatic disease in the kidney, lung, liver, brain, and spleen. Results indicate that tumors treated with Ad-EndoAngio displayed 97% growth reduction compared with controls (p < 0.001). Further, in vivo tumor vascular imaging illustrated a reduction in blood vessel number and lumen diameter size. Kaplan-Meier analysis suggested dramatic survival advantage with Ad-EndoAngio treatment. Importantly, histopathological examination demonstrated marked lung and liver metastasis suppression in the treatment arms. These results suggest that sustained EndoAngio gene therapy has effective antiangiogenic action against human RCC tumors and possesses potential as a novel treatment for metastatic renal cell carcinoma. PMID- 18507515 TI - Preclinical safety and biodistribution of Sindbis virus measles DNA vaccines administered as a single dose or followed by live attenuated measles vaccine in a heterologous prime-boost regimen. AB - Measles still causes considerable morbidity and mortality among infants and young children in developing countries. To develop a new public health tool to reduce this burden, we designed two Sindbis virus replicon vaccines encoding measles virus (MV) hemagglutinin (H) and fusion (F) proteins (pMSIN-H and pMSINHFdU). Our goal is to administer the vaccines to young infants at 6 and 10 weeks of age to prime the immune system to safely and effectively respond to subsequent immunization at age approximately 14 weeks with the licensed attenuated measles vaccine. In preparation for a phase 1 clinical trial, studies of plasmid distribution, integration, and toxicology were performed in rabbits. Biodistribution was assessed after a single DNA immunization delivered intradermally by needle-free injection. Toxicity was assessed using a heterologous prime-boost regimen consisting of a repeat-dose DNA prime followed by a live-attenuated measles vaccine boost. The only vaccine-related adverse effects observed were minimal transient erythema, edema, and inflammation confined to the injection site. Plasmids were detected in the subcutis and muscle at the site of inoculation. A small proportion of animals exhibited plasmids in the regional lymph nodes. There was no evidence of plasmid integration into the host genome. Both Sindbis-based vaccine plasmids were immunogenic in rabbits; pMSIN-H elicited higher virus-neutralizing antibody levels. Both vaccines were shown to be well tolerated and suitable for clinical trials and they are currently being tested in phase 1 studies in young adults. PMID- 18507518 TI - On net reproductive rate and the timing of reproductive output. AB - Abstract: Understanding the relationship between life-history patterns and population growth is central to demographic studies. Here we derive a new method for calculating the timing of reproductive output, from which the generation time and its variance can also be calculated. The method is based on the explicit computation of the net reproductive rate (R0) using a new graphical approach. Using nodding thistle, desert tortoise, creeping aven, and cat's ear as examples, we show how R0 and the timing of reproduction is calculated and interpreted, even in cases with complex life cycles. We show that the explicit R0 formula allows us to explore the effect of all reproductive pathways in the life cycle, something that cannot be done with traditional analysis of the population growth rate (lambda). Additionally, we compare a recently published method for determining population persistence conditions with the condition R0 > 1 and show how the latter is simpler and more easily interpreted biologically. Using our calculation of the timing of reproductive output, we illustrate how this demographic measure can be used to understand the effects of life-history traits on population growth and control. PMID- 18507519 TI - Selection through male function favors smaller floral display size in the common morning glory Ipomoea purpurea (Convolvulaceae). AB - In self-compatible, hermaphroditic plants, display size-the number of flowers open on a plant at one time-is believed to be influenced by trade-offs between increasing geitonogamous selfing and decreasing per-flower pollen export as display size increases. Experimental results presented here indicate that selection through male function favors smaller display sizes in Ipomoea purpurea. In small arrays, plant display size was manipulated experimentally, and female selfing rate, male outcross success, and total male fitness were estimated using genetic markers and likelihood and regression analyses. As would be expected if larger displays experience greater geitonogamy, selfing rate increased with display size. However, the per-flower amount of pollen exported to other plants decreased with display size. The magnitude of this effect is more than sufficient to offset the increase in selfing rate, resulting in reduced per-flower total male fitness with increasing display size. The low values of inbreeding depression previously reported for this species would enhance this effect. PMID- 18507520 TI - Shift in colonial reproductive strategy associated with a tropical-temperate gradient in Rhytidoponera ants. AB - Offspring quality and quantity are subject to a trade-off. Depending on species, ant colonies can produce either numerous winged queens that start new colonies alone (independent colony foundation [ICF]) or one propagule consisting of reproductives helped by workers (fission). Some species do both. In the Rhytidoponera impressa group, colonies with winged queens use ICF, whereas colonies with mated workers use fission. We assessed the effect of latitude and environment on colonial reproduction by collecting 79 colonies along the Australian east coast. A population dynamical model predicted that from tropical north to temperate south, seasonal fluctuations and harsher conditions should lead to a decrease in ICF success (-48%), making fission a suitable alternative. Yet, our empirical data showed that ICF persists at a higher rate than expected, presumably because it allows aerial dispersal, unlike fission. Investigation of colony demography and measurement of lean and fat weights in virgin winged queens and workers showed that, from north to south, colonies increase quality of queens relative to workers (+50%) at the cost of quantity (-86%). This modification limits the decrease in ICF success (-34%). A tremendous range of offspring phenotypes associated with quality-quantity trade-offs make ants competitive in diverse habitats. PMID- 18507521 TI - Long-term efficacy and tolerance of efavirenz- and nevirapine-containing regimens in adult HIV type 1 Senegalese patients. AB - Owing to their low toxicity, low price, and ease of use, efavirenz (EFV) and nevirapine (NVP) are frequently used as part of antiretroviral regimens for AIDS treatment. Several clinical trials have already studied their efficacy and tolerance. However, long-term observations of the effects of these drugs in patients are limited. We used data from a prospective Senegalese cohort to analyze long-term tolerance and efficacy of these two drugs in a low-resources setting. Patients were included if they started their therapy with EFV or NVP. They were censored after treatment discontinuation. The primary endpoint was the time to treatment discontinuation. Secondary endpoints included time to death, time to disease progression, occurrence of severe adverse effects, CD4 cell recovery, and virological response. Confounding factors were controlled using marginal structural models. The median follow-up time in both EFV and NVP arms was 48 months. The hazard ratio (HR) of drug discontinuation in the EFV arm vs. the NVP arm was 0.84 (0.34; 1.87). There was a borderline difference in virological response [HR 1.38 (0.999; 1.89)] but no differences in time to death [HR 1.15 (0.41; 3.24)], time to AIDS progression [HR 1.25 (0.61; 2.58)], or time to increase in CD4 cell count above 500 cells/mm3. Adverse effects were different between NVP and EFV, but long-term tolerance was good for both. This analysis provided further information on long-term tolerance and efficacy of EFV and NVP in a resource-limited setting. PMID- 18507522 TI - Diversity of human immunodeficiency virus type 1 subtypes in Kagera and Kilimanjaro regions, Tanzania. AB - A strategy to prevent the spread of HIV-1 worldwide is complicated by the high genetic diversity of the virus. To gain a better understanding of the HIV-1 genetic diversity in Tanzania, a molecular epidemiological investigation was conducted in Kagera and Kilimanjaro regions. While several studies have addressed HIV-1 subtypes in Tanzania, this is the first study to describe the virus subtypes circulating in Kagera. The Kagera region is the epicenter of the HIV-1 epidemic in Africa, and it was therefore of interest to compare the prevalence of HIV subtypes in this region and Kilimanjaro. Blood samples were obtained from 246 HIV-1-infected pregnant women attending antenatal clinics. Plasma HIV-1 RNA was extracted, amplified, and sequenced in the env C2V3 and/or pol regions from 209 samples. Based on the analysis of env C2V3 and pol sequences, 47.4% had concordant subtypes, 19.1% were discordant indicating recombination, and for 33.5% sequences were obtained for only one region. The distribution HIV-1 subtypes based on the phylogenetic analysis of paired env C2V3/ pol sequences in Kagera region was A/A (27.8%), C/C (29.6%), D/D (16.7%), and unique recombinant forms (25.9%), and in Kilimanjaro region was A/A (32.9%), C/C (25.9%), D/D (10.6%), CRF10_CD (1.2%), and unique recombinant forms (29.4%). The env C2V3 subsubtype A2 and env C2V3/pol CRF10_CD were also observed indicating that these recombinants are circulating in Tanzania. The high diversity of HIV-1 subtypes and the high prevalence of recombinants demonstrated in this study necessitate expanded and continuous monitoring of the epidemic in Tanzania. The trend may have implications for current national control strategies against the HIV-1 epidemic. PMID- 18507523 TI - Plasma HIV RNA decline and emergence of drug resistance mutations among patients with multiple virologic failures receiving resistance testing-guided HAART. AB - Early recognition of virologic failure in patients with extensive drug resistance receiving salvage-HAART is essential to avoid exposure to subinhibitory regimens. We studied plasma viral load (PVL) decline and rates of drug-resistance mutation (DRM) accumulation in such patients. A prospective, 48 week study of 38 heavily pretreated patients receiving genotypic resistance testing (GRT)-guided HAART was conducted. The rate of PVL decline was studied by weekly PVL determinations. To assess DRM accumulation, serial GRTs were performed in all nonresponders (never reaching PVL <50 or two PVLs >50 copies/ml after suppression). Over 48 weeks, 10 patients (26%) were nonresponders. Receiving less then two fully active drugs and having an elevated number of PI and NRTI mutations at baseline were strongly associated with virologic failure. There was no evidence of a difference in the change from baseline PVL to week 1 and 2 between responders and nonresponders. By contrast, PVL reductions from week 2 to week 3 and thereafter were significantly greater for responders (p < 0.01). Among nonresponders, the incidence rates per patient-month (95% CI) of emergent DRM were 0.67 (0.13-1.20), 0.40 (0.00-0.74), and 0.37 (0.00-0.75) at weeks 4, 8, and 24, respectively. Having limited baseline resistance, receiving at least two fully active drugs, and showing constant PVL reductions from week 2 to week 3 and thereafter were predictive of virologic response. In contrast, early changes in PVL levels were not. Virologic failure was associated with detection of emergent DRMs. Virologic rebound in patients on salvage-HAART should be addressed aggressively. PMID- 18507524 TI - Switch from ritonavir-boosted to unboosted atazanavir guided by therapeutic drug monitoring. AB - Plasma concentration of atazanavir (ATV) may be reduced when coadministered with tenofovir (TDF) or proton pump inhibitors. Boosting ATV exposure with ritonavir (r) may make it possible to overcome these drug interactions. However, jaundice and loss of the metabolic advantages of ATV are more frequent using ATV/r than ATV alone. Herein, we assessed whether therapeutic drug monitoring (TDM) could make it possible to identify the subset of patients in whom removal of ritonavir could be attempted without risk of suboptimal plasma ATV exposure and subsequent virological failure. A total of 56 patients with undetectable plasma HIV-RNA under a stable triple regimen containing ATV 300/100 mg qd were switched to ATV 400 mg qd. Plasma ATV concentrations were measured using a reliable high performance liquid chromatography method. Median plasma ATV C(min) fell from 880 to 283 ng/ml (p = 0.03) after removal of ritonavir. While all patients on ATV/r showed ATV plasma concentrations within therapeutic values (IC(min) above 150 ng/ml) before switching, four patients (7%) fell below this threshold after switching to ATV 400 mg qd. However, only one of this group experienced virological failure at week 24 of follow-up. TDF was part of the antiretroviral regimen in all four cases. From a total of 29 (52%) patients on ATV/r showing grade 3-4 hyperbilirubinemia, only 7 (12%) remained on it upon switching to ATV 400 mg qd (p < 0.001). Patients with complete viral suppression under ATV/r 300/100 mg qd may benefit from switching to ATV 400 mg qd guided by TDM, which may make it possible to minimize adverse events without compromising antiviral efficacy in most cases. PMID- 18507525 TI - An open-label safety study of enfuvirtide injection with a needle-free injection device or needle/syringe: the Biojector 2000 Open-label Safety Study (BOSS). AB - Enfuvirtide (ENF) administration by needle/syringe is commonly associated with injection site reactions (ISRs). This study assessed ISRs and participant preference between a needle-free injection device (NFID) and a 27-gauge 1/2-inch needle/syringe (NS). A total of 349 participants with human immunodeficiency virus infection, who had difficulty tolerating long-term administration of ENF by NS, underwent randomization (2:1) to ENF administered twice daily by NFID for 8 weeks, or by NS for 4 weeks followed by NFID for 4 weeks. The objectives of the study were to compare ISRs associated with ENF injection using NFID or NS based on a composite endpoint, ISR incidence/severity, overall ISR scores, and discontinuations. In the NFID group, ISRs improved as the percentage of participants meeting the composite endpoint decreased from baseline (40.1%) to week 4 (25.4%) and remained stable at week 8 (21.2%). In the NS --> NFID group, the percentage meeting the composite endpoint worsened from baseline (36.5%) to week 4 (45.1%), but improved at week 8 (26.1%) after switching. Between participant comparison showed a statistically significant greater improvement from baseline to week 4 in overall ISR score in the NFID group compared to the NS group. Within-participant comparison of the NS --> NFID group showed a significantly greater decrease in overall ISR score from baseline to week 8. In responses to a questionnaire, 87.2% of the participants surveyed preferred the NFID delivery system over NS. NFID is an alternative injection method that may reduce the incidence and severity of treatment-limiting ISRs associated with ENF administration. PMID- 18507527 TI - Study of the genotypic resistant pattern in HIV-infected women and children from rural west Cameroon. AB - The distribution of antiretroviral (ARV) therapy resistance mutations among HIV-1 strains was evaluated in 39 postpartum women, one pregnant woman, and 12 HIV positive babies (seven newborns and five children) living in rural west Cameroon. Thirty-five women and all newborns received a single dose of nevirapine (NVP) to prevent mother-to-child transmission of HIV-1; two women were ARV treated and three were ARV naive. Of the 52 viral strains examined, three were subtype B, 45 were classified into eight HIV-1 non-B subtypes, and four remained unclassifiable. Sequence analysis for genotypic drug resistance in the reverse transcriptase (RT) gene showed the presence of mutations associated with nonnucleoside RT inhibitor resistance in 20% of the samples from NVP-treated women and in 57% of those from treated newborns. Mutations associated with nucleoside RT inhibitors (M184V in one case and V118I in four cases) were found in five samples, despite being derived from ARV-naive patients. As expected, a greater frequency of mutations was found in the protease gene region. Of the sequences analyzed, 79% harbored five to seven specific mutations. The secondary mutations showed the typical protease inhibitor resistance-associated pattern for non-subtype B viruses, M36I being the predominant mutation (92.5% in women, 100% in babies). Other mutations frequently detected were K20I, L63P, H69K, and I13V. These findings confirm that resistance mutations can be detected in ARV-naive patients infected with non-B subtypes and emphasize an urgent need for studies assessing the impact of these mutations on the efficacy of subsequent ARV therapy and on the appearance of drug-resistant strains. PMID- 18507526 TI - Antiretroviral resistance among HIV type 1-infected women first exposed to antiretrovirals during pregnancy: plasma versus PBMCs. AB - Resistance-associated mutations (RAMs) in plasma samples from HIV-1-infected women who received antiretroviral (ARV) prophylaxis during pregnancy was assessed and correlated with the detection of RAMs in peripheral blood mononuclear cells (PMBCs). The study population was composed of HIV-1-infected women enrolled in a prospective cohort study in Latin America and the Caribbean (NISDI Perinatal Study) as of March 1, 2005, who were diagnosed with HIV-1 infection during the current pregnancy, who received ARVs during pregnancy for prevention of mother-to child transmission of HIV-1, and who were followed through at least the 6-12 week postpartum visit. Plasma samples collected at enrollment during pregnancy and at 6-12 weeks postpartum were assayed for RAMs. Plasma results were compared to previously described PBMC results from the same study population. Of 819 enrolled subjects, 197 met the eligibility criteria. Nucleic acid amplification was accomplished in 123 plasma samples at enrollment or 6-12 weeks postpartum, and RAMs were detected in 22 (17.9%; 95%CI: 11.7-25.9%). Previous analyses had demonstrated detection of RAMs in PBMCs in 19 (16.1%). There was high concordance between RAMs detected in plasma and PBMC samples, with only eight discordant pairs. The prevalence of RAMs among these pregnant, HIV-1-infected women is high (15%). Rates of detection of RAMs in plasma and PBMC samples were similar. PMID- 18507528 TI - Short communication: oral lesions in HIV/AIDS patients undergoing HAART including efavirenz. AB - Oral lesions (OL) have an important prognostic value for HIV/AIDS patients. However, the behavior of OL in HIV/AIDS patients undergoing highly active antiretroviral therapy including efavirenz (HAART/EFV) has not been documented. Our objective was to establish the prevalence of OL in HIV/AIDS patients undergoing HAART/EFV and to compare it with the prevalence of OL in patients undergoing antiretroviral therapy including a protease inhibitor (HAART/PI). Seventy-three HIV/AIDS patients undergoing antiretroviral treatment for at least for 6 months at "La Raza" Medical Center's Internal Medicine Unit (IMSS, Mexico City) were included. To detect OL, a detailed examination of oral soft tissues was performed in each patient. Patient records recorded gender, seropositivity time, route of contagion, antiretroviral therapy type and duration, CD4 lymphocyte count/ml, and viral load. Two groups were formed: 38 patients receiving HAART/EFV [two nucleoside analogue reverse transcriptase inhibitors (NARTI) plus efavirenz] and 35 patients receiving HAART/PI (two NARTIs plus one PI). OL prevalence was established in each study group. The Chi-square test was applied (p < 0.05(IC95%)). OL prevalence in the HAART/EFV group (32%) was lower (p < 0.007) than in the HAART/PI group (63%). Candidosis was the most prevalent OL in both groups. Herpes labialis, HIV-associated necrotizing periodontitis, xerostomia, hairy leukoplakia, and nonspecific oral sores were identified. The highest prevalence for all OL was found in the HAART/PI group. These findings suggest that HIV/AIDS patients undergoing HAART/EFV show a lower prevalence of oral lesions than patients undergoing HAART/PI. PMID- 18507529 TI - Interarm blood pressure differences in the women's interagency HIV study. AB - Hypertension has been reported in 8-32% of HIV-infected individuals. Large interarm blood pressure differences (IABPD) may cause misclassification of blood pressure (BP) status. The objectives of this study were to determine the magnitude and factors associated with IABPD in HIV-infected women and uninfected controls. Using automated devices, two BP recordings were measured and averaged from each arm in Brooklyn enrollees of the Women's Interagency HIV Study. Absolute IABPD was calculated for each patient. Among 335 subjects, 238 were HIV infected and 97 were uninfected. Mean systolic and diastolic IABPD were 6 +/- 5 mm Hg and 4 +/- 3 mm Hg, respectively. Twenty-six percent of subjects had systolic IABPD >10 mm Hg and 6% had systolic IABPD >20 mm Hg. Fifteen percent of subjects had diastolic IABPD >10 mm Hg. Interarm BP differences were not associated with HIV serostatus, CD4(+) cell count, and use of highly active antiretroviral therapy. Systolic IABPD >20 mm Hg was associated with obesity (ORadj 5.37, 95% CI 1.47, 19.65), and LDL cholesterol above 160 (ORadj 9.12, 95% CI 2.53, 32.88). Right arm BP measurement resulted in 10% of subjects with high/uncontrolled BP. Bilateral arm BP measurement increased the yield to 15% (p < 0.001). In conclusion, systolic and diastolic IABPD are common and appear to be of clinically important magnitude. Systolic IABPD are related to cardiovascular risk factors but not to HIV-related factors. Bilateral BP determination is important to detect and manage hypertension as well as for accurate cardiovascular risk assessment. PMID- 18507530 TI - Genotypic characterization and comparison of full-length envelope glycoproteins from South African HIV type 1 subtype C primary isolates that utilize CCR5 and/or CXCR4. AB - CCR5 has preferentially been used by all circulating HIV-1 subtype C viruses for cell entry. Recently, we reported the highest proportion of CXCR4-utilizing primary isolates among a cohort of 20 South African AIDS patients. This study describes and compares the Env genotypic characteristics from these 20 HIV-1 subtype C (and unique CD recombinant) primary isolates. Fourteen primary isolates utilized CCR5, four (including the CD recombinant) used CXCR4, and two were dual tropic. Extensive analysis and comparison of important structural motifs such as the N-linked glycosylation sites, signal sequences, CD4-binding sites, variable loops, cleavage sites, known neutralizing antibody and small molecule inhibitor binding sites confirmed that other than the expected differences in the V3 loop, no sequence motifs distinguished between R5 and X4 tropism. Further correlation of the env genotype to functionally relevant motifs is necessary to elucidate the relationship between biologically and immunologically relevant sites and aid vaccine and novel drug design. PMID- 18507531 TI - Antibodies against PsrP, a novel Streptococcus pneumoniae adhesin, block adhesion and protect mice against pneumococcal challenge. AB - Pneumococcal serine-rich repeat protein (PsrP) is a putative adhesin encoded in the Streptococcus pneumoniae pathogenicity island psrP-secY2A2. Challenge of mice with serotype 4, strain TIGR4, and the isogenic mutants T4DeltapsrP and T4DeltapsrP-secY2A2 determined that PsrP was required for bacterial persistence in the lungs but not for colonization in the nasopharynx or replication in the bloodstream during sepsis. In vitro experiments corroborated this anatomical site specific role; psrP mutants failed to bind to A549 and LA-4 lung cells, yet adhered normally to human nasopharyngeal epithelial cells and to cells from human and rodent capillary endothelial cell lines. We determined that the amino terminus of PsrP mediated adhesion. Microspheres coated with recombinant PsrP(SRR1-BR) (rPsrP(SRR1-BR)) adhered to A549 cells, and moreover, preincubation of cells with rPsrP(SRR1-BR) inhibited TIGR4 adhesion in vitro. Antibodies against rPsrP(SRR1-BR) also neutralized PsrP function; antiserum against rPsrP(SRR1-BR) blocked TIGR4 adhesion in vitro and, following passive immunization, it protected mice against challenge. We conclude that PsrP is an adhesin required for bacterial persistence in the lungs and that rPsrP(SRR1-BR) is a protective antigen. PMID- 18507532 TI - Beta-adrenergic stimulation enhances the heat-shock response in fish. AB - We have taken advantage of the unique properties of nucleated rainbow trout (Oncorhynchus mykiss) red blood cells (rbcs) to demonstrate that beta-adrenergic stimulation with the agonist, isoproterenol, significantly enhanced the heat induced induction of heat-shock proteins (Hsps) in trout rbcs without affecting hsp expression on its own. Furthermore, this beta-adrenergic potentiation of hsp expression occurred only at physiologically relevant concentrations of adrenergic stimulation. In further experiments, we found that adrenaline increased 100-fold and noradrenaline increased 50-fold in trout after a 1-h heat shock at 25 degrees C, approximately 12 degrees C above acclimation temperature. This is the first time the adrenergic heat-shock response has been described for a temperate fish species. We conclude that beta-adrenergic stimulation enhances hsp expression in trout rbcs following heat stress, indicating physiological regulation of the cellular stress response in fish. PMID- 18507533 TI - Thermal biology, torpor, and activity in free-living mulgaras in arid zone Australia during the winter reproductive season. AB - Little is known about the energy conservation strategies of free-ranging marsupials living in resource-poor Australian deserts. We studied activity patterns and torpor of free-living mulgaras (Dasycercus blythi) in arid central Australia during the winter of 2006. Mulgaras are small (approximately 75 g), nocturnal, insectivorous marsupials, with a patchy distribution in hummock grasslands. Mulgaras (six males, three females) were implanted intraperitoneally with temperature-sensitive transmitters and monitored for 6-55 d. Temperature profiles for different microhabitats and the thermal properties of soil and a number of burrows were also measured. Air temperature ranged from -3 degrees C at night to 30 degrees C during the day. Although burrows buffered temperature extremes, the thermal diffusivity of the sandy soil was high, and many burrows were shallow. Hence, soil and burrow temperatures averaged about 15 degrees C. The activity of mulgaras was often restricted to a few hours after sunset, before they retired into their burrows. Mulgaras employed torpor frequently, often entering torpor during the night and arousing around midday, with arousals occurring later on cooler days. Shallow burrows allowed cooling below mean T(soil). Consequently, body temperatures as low as 10.8 degrees C were observed. The longest torpor bout was 20.8 h. Torpor patterns changed seasonally and differed between males and females. From June to August, females entered torpor almost daily despite mating and gestation, but from the end of the gestation period onward, they remained normothermic. In contrast, males showed only shallow and short torpor during the mating season, but from mid-July, a transition to more frequent and deeper torpor resembling that of females was observed. Apparently, in both sexes, the reproductive effort entails energetic costs, but torpor, as an energy-saving mechanism, and reproduction are not exclusive in mulgaras. In a resource-poor environment during the least productive part of the year, frequent torpor seems to provide the means to compensate for the increased energetic costs associated with reproduction. PMID- 18507534 TI - Association between both lipid and protein oxidation and the risk of fatal or non fatal coronary heart disease in a human population. AB - The role of oxidative damage in the aetiology of coronary disease remains controversial, as clinical trials investigating the effect of antioxidants have not generally been positive. In the present study, 227 coronary cases, identified from a cohort study, were matched, by age and gender, with 420 controls in a nested case-control design. Stored plasma samples were analysed for F2 isoprostanes by stable isotope dilution MS, and specifically oxidized forms of apoA-I (apolipoprotein A-I) by HPLC of HDL (high-density lipoprotein). Median values of F2-isoprostanes were higher in plasma samples that contained oxidized apoA-I compared with samples with undetectable oxidized apoA-I (1542 compared with 1165 pmol/l). F2-Isoprostanes were significantly correlated with variants of non-oxidized apoA-II (r=-0.15) and were associated with HDL-cholesterol (P<0.0001). F2-Isoprostanes in cases (median, 1146 pmol/l) were not different from controls (1250 pmol/l); the odds ratio (95% confidence interval) for a 1 S.D. increase in F2-isoprostanes was 1.08 (0.91-1.29). Similarly, there was no independent association between the presence of oxidized apoA-I, detected in approx. 20% of the samples, and coronary risk. In conclusion, we found no evidence of associations between markers of lipid (F2-isoprostanes) and protein (oxidized apoA-I) oxidation and the risk of fatal or non-fatal coronary heart disease in a general population. This may be due to a true lack of association or insufficient power. PMID- 18507535 TI - The AHA Scientific Statement of MRI in patients with devices: neat, but incomplete. Unwise and unsupported. PMID- 18507536 TI - Segmental pulmonary vein ablation: success rates with and without exclusion of areas adjacent to the esophagus. AB - BACKGROUND: Catheter ablation has become the first line of therapy in patients with symptomatic recurrent, drug-refractory atrial fibrillation (AF). The occurrence of an atrioesophageal fistula is a rare but serious complication after AF-ablation procedures. This risk is even present during segmental pulmonary vein (PV) ablation procedures because the esophagus does frequently have a very close anatomical relationship to the right or left PV ostia. The aim of the present study was to analyze whether the exclusion of areas adjacent to the esophagus does have a significant effect on the success rates after segmental pulmonary vein ablation procedures. METHODS: Forty-three consecutive patients with symptomatic paroxysmal AF were enrolled in this study. In all patients, a segmental PV ablation procedure was performed. The procedures were facilitated by a 3D real-time visualization of the circumferential mapping catheter placed in the pulmonary veins using the NavX system (St. Jude Medical, St. Paul, MN, USA; open irrigated tip ablation catheter; 43 degrees C; 30 W). In 21 patients, a complete ostial PV isolation was attempted regardless of the anatomical relationship between the ablation sites and the esophagus (group A). In the remaining 22 patients, the esophagus was marked by a stomach tube and areas adjacent to the esophagus were excluded from the ablation procedure (group B). After discharge, patients were scheduled for repeated visits at the arrhythmia clinic at 1, 3, and 6 months after the ablation procedure. RESULTS: The segmental pulmonary vein ablation procedure could be performed as planned in all patients. In group A, all pulmonary veins could be isolated successfully in 14 out of 21 patients (67%). A mean number of 3.7 pulmonary veins (SD +/- 0.5 PVs) were isolated per patient. The main reasons for an incomplete PV isolation were: small diameter of the PVs, side branches close to the ostium, or poorly accessible PV ostia. In group B, all PVs could be isolated successfully in only 12 out of 22 patients (55%; P = 0.54). A mean number of 3.2 PVs (SD +/- 0.9 PVs) were isolated per patient (P = 0.05). This was mostly due to a close anatomical relationship to the esophagus. The ablation strategy had to be modified in 16/22 patients in group B because of a close anatomical relationship between the left (n = 10) or right (n = 6) PV ostia and the esophagus. After 3 months, the percentage of patients free from an AF recurrence was not significantly different between the two groups (90% vs 95%; P = 0.61). After 6 months, there was no significant difference between the success rates either (81% vs 82%; P = 1.0). There were no major complications in both groups. CONCLUSIONS: The exclusion of areas adjacent to the esophagus results in a moderately higher percentage of incompletely isolated PVs. However, it does not have a significant effect on the AF recurrence rate during short-term and mid-term follow-up. PMID- 18507537 TI - Effects of the location of myocardial infarction on the spectral characteristics of ventricular fibrillation. AB - BACKGROUND: The location of the myocardial infarction (MI) might modify the spectral characteristics of ventricular fibrillation (VF) in humans. OBJECTIVE: To evaluate the effect of the location of the infarcted area on the spectral parameters of VF. METHODS: Patients with chronic MI (29 anterior, 32 inferior) and induced VF during cardioverter defibrillator implant were retrospectively studied. Dominant frequency (f(d)), organization index (OI), and power of the harmonic peaks were calculated in the device-stored electrograms (EGM) during sinus rhythm (SR) and VF. RESULTS: The f(d) of the VF was not affected by the left ventricular ejection fraction (LVEF) or the MI location (anterior: 4.54 +/- 0.74 Hz, inferior: 4.77 +/- 0.48 Hz, n.s.). The OI was also similar in both groups. However, in patients with inferior MIs, normalized peak power at f(d) was higher (118.3 +/- 18.5 vs 100.6 +/- 28.2, P < 0.01) and the normalized peak power of the harmonics was lower than in the anterior MI group. The analysis of EGM during SR showed similar results. The size of the necrotic area and its distance to the recording electrode might partially explain these results. CONCLUSION: In our series, the spectral characteristics of the EGMs during VF showed significant differences depending on the MI localization. A higher fraction of energy (in the low-frequency region) was seen in inferior MIs, whereas the peak power at the harmonics increased in anterior MIs. A similar effect was seen during SR and VF, suggesting that it is caused by local electrophysiology abnormalities induced by the MI rather than by different intrinsic characteristics of the VF. PMID- 18507538 TI - Organized incessant atrial arrhythmias in the setting of severe, isolated biatrial scarring. AB - INTRODUCTION: Diffuse transmural fibrosis and scarring limited to the area without atrial dilation or significant structural heart or other systemic disease has not been reported. We present three cases of a syndrome characterized by refractory organized atrial arrhythmias, diffuse atrial scarring with electrical silence, and mechanical paralysis in the absence of atrial dilation or any systemic or neurodegenerative disorders. METHODS: Patients referred for electrophysiology study of atrial arrhythmias were included. Electroanatomic mapping with the Carto system (Biosense Webster, Diamond Bar, CA, USA) and magnetic resonance imaging (MRI) with scar sequencing were performed. RESULTS: There was no family or personal history of cardiac, muscular, or developmental diseases. All patients had organized atrial arrhythmias. Echocardiograms showed atrial standstill with normal atrial and ventricular dimensions. No other structural abnormalities were noted. Carto mapping revealed severe biatrial diffuse scarring. The left atrial (LA) was less affected than the right atrial (RA). MRI findings confirmed biatrial scarring. During tachycardia, islands of dissociated electrical activity could be seen in the right atria. Entrainment mapping was not performed in the atria as high-output pacing could not capture the atria. Coronary sinus entrainment demonstrated the coronary sinus(CS) not to be critical to the tachycardia. Ablation was targeted toward channels of low voltage but was not successful in any cases. All required atrioventricular (AV) nodal ablation with pacing. CONCLUSION: An association between biatrial cardiomyopathy and scarring with normal atrial dimensions has been described. Since severe scarring has not been reported with organized arrhythmias this may represent a new syndrome. PMID- 18507539 TI - Quality of ECG monitoring with a miniature ECG recorder. AB - BACKGROUND: Long-term monitoring of patients at risk of arrhythmias would benefit from a miniaturized device. This study evaluated the quality of electrocardiogram (ECG) signals recorded by a miniaturized ECG recorder. METHODS: ECG data were concurrently recorded with an investigational device (Philips Medical Systems, Seattle, WA, USA) and a Holter recorder from patients with cardiac-related symptoms at the emergency department of a tertiary care university hospital. The device was attached in one of four locations (L1: below left clavicle, L2: midsternal, L3: below left breast, L4: left anterior axillary line in 5th intercostal space). Selected ECG strips were analyzed for atrial activity, QRS width, and basic rhythm by two physicians. Patients were divided into groups based on their number of strips (A 2-5, B 6-9, C 10-21) for descriptive presentation of the data. For statistical analyses, nonlinear mixed-effects models were used. RESULTS: In 90 patients, 574 pairs of strips were analyzed. In L1 (n = 37), agreement between the device and the Holter recorder on presence of P wave was 58-80% in groups A-C and in L2 (n = 28):94-98%, L3 (n = 12):30-72%, and L4 (n = 13):0-70%; on PR interval in L1:58-80%, L2:94-98%, L3:30-62%, and L4:0-70%; on rhythm in L1:56-79%, L2:94-98%, L3:30-62%, and L4:0-70%; on QRS width in L1:86-100%, L2:88-98%, L3:100%, and L4:96-100%. The agreement of L2 in the parameters P wave, PR interval, and rhythm diagnosis was better than in the other locations (P < 0.01). CONCLUSIONS: A miniature ECG monitor provided accurate assessment of atrial beats and rhythm diagnosis at a midsternal location and of QRS width at all locations. PMID- 18507540 TI - AV junctional ablation allowing more effective delivery of cardiac resynchronization therapy in patients with intra- and interatrial conduction delay. AB - We report two patients with cardiac resynchronization therapy (CRT) devices and evidence of refractory heart failure in whom impaired intraatrial conduction in one patient, and interatrial conduction in the other, prohibited optimization of the atrioventricular (AV) timing sequence. The patient with intraatrial conduction delay exhibited late right atrial sensing and latency during right atrial pacing that required programming of a short-sensed AV delay and long-paced AV delay (wide differential AV delay). In both patients AV junctional ablation and echocardiography-guided device optimization significantly improved heart failure. PMID- 18507541 TI - Automatic determination of timing intervals for upper limit of vulnerability using ICD electrograms. AB - BACKGROUND: Implantable cardioverter defibrillator (ICD) implant testing based on the upper limit of vulnerability, or vulnerability testing, permits assessment of defibrillation safety margins without inducing ventricular fibrillation (VF) in most patients. Vulnerability testing requires that T-wave shocks be timed at the most vulnerable intervals of the cardiac cycle, defined as intervals at which the strongest shock induces VF. Our goal was to develop and test an automated method to select these timing intervals using ICD intracardiac electrograms (EGMs). METHODS: At ICD implant in 22 patients, we determined the range of the most vulnerable intervals by scanning the T wave with shocks. Simultaneously, EGMs were recorded for 351 pacing sequences used for measurement of timing intervals or T-wave shocks. EGMs were analyzed off-line using a novel automated method to identify a stable point near the maximum slope of the T wave in the far-field (shock) EGM. Fiducial timing points based both on the EGM and on the electrocardiogram (ECG) were used to predict the most vulnerable intervals. We compared the predicted most vulnerable to the measured most vulnerable intervals determined by T-shock scans. RESULTS: Automatically determined timing points from EGMs and operator-determined timing points from the surface ECG had comparable accuracy in identifying the measured most vulnerable intervals (91% EGM vs 86% ECG, P = NS). CONCLUSIONS: An automated method based on ICD EGMs identifies the most vulnerable intervals with accuracy comparable to the operator-performed, clinical method based on the surface ECG. This EGM method can be implemented efficiently in an ICD to automate vulnerability testing. PMID- 18507542 TI - Time course of effects of cardiac resynchronization therapy in chronic heart failure: benefits in patients with preserved exercise capacity. AB - OBJECTIVES: To assess in patients with chronic heart failure the effect of cardiac resynchronization therapy (CRT) over 12 months' follow-up the time course of the changes in functional and neurohormonal indices and to identify responders to CRT. METHODS: Eighty-nine patients (74.1 +/- 1 years, left ventricular ejection fraction [LVEF] < 35%), QRS complex duration >150 ms, in stable New York Heart Association (NYHA) class III or IV on optimal medical treatment were prospectively randomized either in a control (n = 45) or CRT (n = 44) group and underwent clinical evaluation, cardiopulmonary exercise testing (CPET), 2D-Echo, heart rate variability (HRV), carotid baroreflex (BRS), and BNP assessments before and at 6- and 12-month follow-up. RESULTS: In the CRT group, improvement of cardiac indices and BNP concentration were evident at medium term (over 6 months) follow-up, and these changes persisted on a longer term (12 months) (all P < 0.05). Instead CPET indices and NYHA class improved after 12 months associated with restoration of HRV and BRS (all P < 0.05). We identified 26 responders to CRT according to changes in LVEF and diameters. Responders presented less depressed hemodynamic (LVEF 25 +/- 1.0 vs 22 +/- 0.1%), functional (peak VO(2) 10.2 +/- 0.2 vs 6.9 +/- 0.3 ml/kg/min), and neurohormonal indices (HRV 203.6 +/- 15.7 vs 147.6 +/- 10.ms, BRS 4.9 +/- 0.2 vs 3.6 +/- 0.3 ms/mmHg) (all P < 0.05). In the multivariate analysis, peak VO(2) was the strongest predictor of responders. CONCLUSIONS: Improvement in functional status is associated with restoration of neurohormonal reflex control at medium term. Less depressed functional status (peak VO(2)) was the strongest predictor of responders to CRT. PMID- 18507543 TI - The COGNITION study rationale and design: influence of closed loop stimulation on cognitive performance in pacemaker patients. AB - BACKGROUND: Several studies showed the beneficial effect of pacemaker implantation on cognitive performance in patients with bradycardia. But it has never been investigated if patients with chronotropic incompetence may improve their cognitive performance if treated by a rate-adaptive system reacting to mental stress in comparison to the most frequently used accelerometer-driven pacing. METHODS: The randomized, single-blind, multicenter COGNITION study evaluates if closed loop stimulation (CLS) offers incremental benefit in the speed of cognitive performance and the overall well-being of elderly patients with bradycardia compared with accelerometer-based pacing. Four hundred chronotropically incompetent patients older than 55 years will be randomized 3-6 weeks after implantation to CLS or accelerometer sensor. Follow-up visits are performed after 12 and 24 months. The speed of cognitive performance, which is the underlying function influencing all other aspects of cognitive performance, will be assessed by the number connection test, a standardized psychometric test for the elderly. Secondary endpoints include patient self-assessment of different aspects of health (by visual analogue scales), quality of life (by SF-8 health survey), the incidence of atrial fibrillation (episodes lasting for longer than 24 hours), and the frequency of serious adverse events. CONCLUSION: In the ongoing COGNITION study, we aim at long-term comparison of two rate-adaptive systems, focusing on the cognitive performance of the patients, which was neglected in the past evaluation of pacemaker sensors. PMID- 18507544 TI - Biventricular pacing attenuates T-wave alternans and T-wave amplitude compared to other pacing modes. AB - BACKGROUND: The impact of altered ventricular activation, including biventricular (BV) pacing, on T-wave alternans (TWA) and arrhythmic substrates is unclear. We studied how differing ventricular activation sequence alters TWA; the interval from peak-to-end of the T-wave (TpTe) and T-wave amplitude during right (RV), left (LV), and biventricular (BV) pacing; and right atrial (RA) pacing in patients with preexisting conduction delay. METHODS AND RESULTS: We measured TWA during RA, RV, LV, and BV pacing in 33 patients receiving cardiac resynchronization-therapy-defibrillators. TWA magnitude (V(alt)) was lower during BV than RV (P < 0.01), RA (P < 0.01), or LV pacing. As a result, BV-TWA was more often negative than RV-TWA (P < 0.01), LV-TWA, and RA-TWA, particularly when discordant between pacing modes (P < 0.01). Overall, 83% of TWA recordings were abnormal (25% indeterminate), and 17% negative. BV pacing reduced T-wave amplitude (P < 0.05) and TpTe (P < 0.005) compared to RV pacing and LV pacing (P < 0.05; P < 0.005 respectively). Notably, TWA magnitude varied linearly with T wave amplitude for all pacing modes (P < 0.001). Over 410 +/- 252 days' follow up, RV-TWA predicted the combined endpoint of death and ICD therapy with 86% negative predictive value (P < 0.05). BV-TWA, RA-TWA, and other repolarization indices were not predictive. CONCLUSIONS: BV pacing attenuates TWA in tandem with reduced T-wave magnitude. In these patients with baseline QRS prolongation, RV TWA predicted events more effectively than BV-TWA and RA-TWA. Further studies are required to understand how altered ventricular activation influences repolarization dynamics and arrhythmic tendency. PMID- 18507545 TI - Does size really matter? A comparison of the Riata lead family based on size and its relation to performance. AB - BACKGROUND: Recently, the performance and safety of smaller diameter implantable cardioverter defibrillator (ICD) leads has been questioned. The purpose of this analysis was to determine the impact of size on lead performance and perforation rates by comparing the performance of 7 French (7F) and 8 French (8F) leads with similar design characteristics implanted by a single operator. METHODS: Patients implanted with a Riata 1580 (8F) or 7000 (7F) series leads (St. Jude Medical, Sylmar, CA, USA) over a 2-year period were evaluated to compare performance and perforation rates. RESULTS: There were 357 Riata 8F leads and 357 Riata 7F leads implanted in 714 patients. Follow-up ranged from 1 to 24 months. The 8F leads were implanted in the right ventricular apex more often than were 7F leads (129 or 37% vs 72 or 20%, P < 0.0001). Oversensing that did not result in therapy occurred in 2 pts (0.56%) with 8F leads and 1 pt. (0.28%) with a 7F lead (P = 0.56). Oversensing with therapy occurred once in both groups (0.28%, P = NS). One perforation occurred in each group (0.28%, P = NS). Both occurred in leads that were implanted in the right ventricular apex (P = 0.02). CONCLUSIONS: The performance of St. Jude Medical 7F and 8F Riata leads was similar. The incidence of lead-related adverse events was within or below the low end of published acceptable ranges for ICD lead perforation and sensing anomalies. Perforations were less likely to occur in leads that were implanted in nonapical positions. PMID- 18507546 TI - Effects of scatter radiation on ICD and CRT function. AB - BACKGROUND: Effects of direct radiation on implantable cardiac devices have been well studied. However, the effects of scatter radiation are not as clear. Recommendations on management of patients with implantable cardiac devices undergoing radiotherapy are based on limited studies mostly involving pacemakers. We sought to elucidate the effects of scatter radiation on implantable cardioverter-defibrillators (ICDs) and cardiac resynchronization therapy (CRT) ICDs. METHODS: We exposed 12 ICDs and eight CRT-ICDs to 400 cGy of scatter radiation from a 6-MV photon beam. Devices were programmed with nominal parameters and interrogated prior to radiation, after each fraction, upon completion of the radiation course and again 1 week later. A retrospective review of patients undergoing radiotherapy at the Mayo Clinic-Rochester between 2002 and 2007 in whom the device was outside the radiation field was also performed. There were 13 patients with devices undergoing radiotherapy during this time period, 12 of whom were interrogated prior to and after radiation. RESULTS: Interrogation reports were reviewed for device reset or parameter changes. There was no evidence of reset or malfunction during or after radiation. Also, no episodes of device reset, inappropriate sensing or therapy, or changes in programmed parameters were found in our review of patients undergoing radiotherapy. CONCLUSIONS: Device reset or malfunction associated with scatter radiation likely represents an unpredictable, rare occurrence. While we see no clear contraindication to radiotherapy in patients with ICDs or CRT-ICDs, precautions should be taken to avoid direct radiation exposure and to closely evaluate patient outcomes before and after the radiation course. PMID- 18507547 TI - Pacemaker malfunction due to atrial and ventricular leads switched in the header: two faces of the same mistake? PMID- 18507548 TI - Extraction of transvenous pacing and ICD leads. AB - The emergence of pacing and implantable cardioverter-defibrillator (ICD) systems, along with expanding indications of these devices (e.g., cardiac resynchronization therapy and sudden cardiac death prevention), increasing infection rates, and device recalls have created the need for removing and upgrading these systems due to various reasons. Removing the pulse generator of a system is generally uncomplicated. Chronically implanted transvenous leads, however, adhere to the venous endothelium and endocardial tissues over time due to fibrosis. Removal of such leads can be a significantly complex procedure requiring tools and techniques that free the lead at fibrotic binding sites. In this article, the state-of-the-art tools and techniques that provide a systematic approach to consistently and safely extract these devices will be reviewed. PMID- 18507549 TI - Paroxysmal atrial fibrillation ablation with the multipolar mapping and ablation catheter (Mesh-Bard). AB - Pulmonary veins (PVs) ablation is a valid treatment option for atrial fibrillation. The standard approach for PVs isolation usually requires two catheters or an electroanatomical reconstruction of the left atrium. We describe our initial experience with a single device for mapping and ablating in a patient referred to our center for the relapsing of atrial fibrillation 3 years after a previous ablation procedure. The newly available catheter MESH was safe and effective to quickly isolate two reconnected PVs. PMID- 18507550 TI - Incessant non-reentrant supraventricular tachycardia interrupted by apparent simultaneous wenckebach sequences along both nodal pathways. AB - We report a new case of non-reentrant supraventricular tachycardia, associated with tachycardia-induced cardiomyopathy fully reversible after radiofrequency (RF) ablation, together with striking features of apparent concomitant Mobitz type 1 atrioventricular (AV) block in both AV node pathways. Further analysis of the conduction patterns during the incessant non-reentrant tachycardia raised unresolved hypothesis about the involved mechanisms and further interrogations on AV node physiology. PMID- 18507551 TI - Brugada-like ST-segment abnormalities associated with myocardial involvement of hematologic diseases. AB - This is a report of Brugada-like ST-segment abnormalities related to acute myocarditis associated with hematologic disorders. Electrocardiographic (ECG) pattern of ST-segment elevation in the right precordial leads mimicking Brugada syndrome may relate to pathological abnormalities due to hematologic disorders that may have genetic, infective, or inflammatory origins. We describe two cases of myocardial involvement of hemotologic disorders, manifested with Brugada-like ECG findings. PMID- 18507552 TI - Ranolozine-induced suppression of ventricular tachycardia in a patient with nonischemic cardiomyopathy: a case report. AB - Ranolazine is an antianginal agent, which inhibits the abnormal inward Na(+) current and by this inhibition decreases diastolic cardiomyocyte calcium levels and improves electrical stability. Ranolazine has also been shown to be a potent inhibitor of after depolarizations and triggered activity. In this case report, we describe the dramatic antiarrhythmic effects of ranolazine in a patient with nonischemic cardiomyopathy who had malignant ventricular tachycardia. Further research on the antiarrhythmic properties of ranolazine appears warranted. PMID- 18507553 TI - B-Cell lymphoproliferative disorder of an ICD pocket: a diagnostic puzzle in an immunosuppressed patient. AB - We present a case of a patient with lymphoma in an ICD pocket in the setting of posttransplant immune suppression. Infection of the ICD system was suspected and the correct diagnosis was established by biopsy. PMID- 18507554 TI - Concomitant-acquired Long QT and Brugada syndromes associated with indapamide induced hypokalemia and hyponatremia. AB - Electrolyte disturbances are known to cause acquired Long QT syndrome (LQTS) and Brugada syndrome. While a reduction in INa due to SCN5A mutation is implicated as the underlying mechanism in Brugada syndrome, hyponatremia, which can give rise to a reduced INa, has never been reported in literature as a cause or precipitating factor in this syndrome. We detailed a case in which concomitant acquired LQTS and Brugada syndrome were associated with severe hypokalemia and hyponatremia following indapamide use for treatment of hypertension and highlighted the potential role of hyponatremia in the pathogenesis of the acquired form of Brugada syndrome. PMID- 18507555 TI - Precipitation of ventricular fibrillation by intravenous diltiazem and metoprolol in a young patient with occult Wolff-Parkinson-White syndrome. AB - We report the case of a young man who presented with a rapid, narrow-complex atrial fibrillation. A few hours after being administered intravenous metoprolol and diltiazem for rate control, he developed intermittent ventricular preexcitation on the electrocardiogram (ECG) and experienced ventricular fibrillation, from which he was successfully defibrillated. A subsequent electrocardiogram in sinus rhythm demonstrated previously unknown Wolff-Parkinson White pattern. A left lateral accessory pathway was successfully ablated. Wolff Parkinson-White syndrome should be included in the differential diagnosis when a young patient presents with atrial fibrillation, even if the ventricular complexes on the ECG are not preexcited. PMID- 18507556 TI - Partial inhibition of ongoing antitachycardia pacing sequence due to T-wave oversensing. AB - We describe a case of partial inhibition of ongoing antitachycardia pacing scheme in an implantable cardioverter defibrillator (ICD) recipient with heart failure of ischemic etiology. The cause of inhibition was related to intermittent oversensing of paced T wave and sensing of intrinsic ventricular activation. In our effort to solve this problem, we reprogrammed the paced ventricular blanking period to a higher level. PMID- 18507557 TI - Short PR interval in the absence of preexcitation: a characteristic finding in a patient with Fabry disease. AB - Fabry disease (FD) is a rare lysosomal storage disease characterized by intracellular deposition of glycosphingolipids that can involve various organs including the heart. Here, we report the finding of a short PR interval in a patient with FD and recurrent syncope associated with short atrial-His (A-H) and His-ventricular (H-V) intervals in the absence of preexcitation. While electrocardiographic abnormalities are not uncommon among FD patients, there have been few reports on electrophysiological studies performed on these patients. The exact mechanism for the short A-H and H-V intervals remains unclear, but they may result from accelerated conduction in the AV node-His-Purkinje system. PMID- 18507558 TI - More late perforations with the Riata defibrillator lead from a high-volume center: an update on the numbers. PMID- 18507559 TI - Delayed defibrillator lead perforation. PMID- 18507560 TI - Current of injury (COI) pattern recorded from catheter delivered active fixation pacing leads. PMID- 18507563 TI - Effective strategies for implementing a multicenter international clinical trial. AB - PURPOSE: International collaboration in research is essential in order to improve worldwide health. The purpose of this paper is to describe strategies used to administer an international multicenter trial to assess the effectiveness of a nursing educational intervention. DESIGN: The study design was a two-group randomized multicenter international clinical trial conducted to determine whether a brief education and counselling intervention delivered by a nurse could reduce prehospital delay in the event of symptoms suggestive of acute coronary syndrome (ACS) in patients previously diagnosed with cardiovascular disease. METHOD: A flexible but well-defined project structure showed intervention consistency in five sites among three countries and included experienced project coordinators, multidimensional communication methods, strategies to optimize intervention fidelity, site-specific recruitment and retention techniques, centralized data management, and consideration of ethical and budgetary requirements. FINDINGS: Staff at five sites enrolled 3,522 participants from three countries and achieved 80% follow-up obtained at both 12 and 24 months. CONCLUSION: Multidimensional approaches to maintain consistency across study sites, while allowing flexibility to meet local expectations and needs, contributed to the success of this trial. CLINICAL RELEVANCE: In order to support appropriate development of an evidence base for practice, nursing interventions should be tested in multiple settings. A range of strategies is described in this paper that proved effective in conducting a multicenter international trial. PMID- 18507564 TI - Interaction of duration of homelessness and gender on adolescent sexual health indicators. AB - PURPOSE: The purpose of this analysis was to determine the effects of duration of homelessness and gender on personal and social resources, cognitive-perceptual factors, and sexual health behaviors among homeless youth. DESIGN: Cross sectional analysis of data collected at baseline from 461 homeless adolescents who participated in a sexual health intervention study was done. METHOD: Data were collected via laptop computers from homeless adolescents (mean age=19.52+1.91 years) in both comparison and intervention groups before the initiation of the intervention. FINDINGS: Significant interaction effects were found for personal and social resources F (4, 426)=2.83, p<.05. Male participants who had been homeless<6 months had significantly higher scores on social connectedness than did male participants who were homeless>1 year. Univariate analysis of variance (ANOVA) indicated that both boys and girls who had been homeless>1 year had greater AIDS knowledge, F (1, 441)=7.91, p<.01, reported significantly more sexual risk-taking behaviors, F (1, 396)=9.93, p<.05, and engaged in fewer safe-sex behaviors, F (1, 396)=12.05, p<.05, than did those who had been homeless<6 months. Univariate ANOVA indicated that female participants had significantly lower levels of perceived health status, F (1, 429)=12.08, p<.01, significantly greater sexual self-care behaviors, F (1, 396)=16.29, p<.01, and significantly higher levels of assertive communication F (1, 396)=4.03, p<.05 than did male participants, regardless of duration of homelessness. CONCLUSIONS: The duration of homelessness and gender has both direct and interaction effects on cognitive-perceptual and behavioral outcomes associated with sexual health. CLINICAL RELEVANCE: Nurses and other healthcare providers working with homeless youth recognize the need to develop brief interventions that address health-risk behaviors. Findings from this study indicate that gender-specific interventions should be provided to youth soon after they become homeless. PMID- 18507565 TI - Testing a bioecological model to examine social support in postpartum adolescents. AB - PURPOSE: The purpose was twofold and included examining a bioecological model as a framework to describe social support in postpartum adolescents. The second purpose was to determine the relationship between a comprehensive view of the context of social support and symptoms of depression. DESIGN: Cross-sectional design with convenience sampling (n=85) of adolescents at 4-6 weeks postpartum, recruited from two community hospitals. METHODS: Approval was received from the university's IRB (institutional review board), each recruitment site, the adolescent mothers, and their parents or guardians. Data were collected by a research assistant during home visits using a battery of self-report instruments to measure macro, meso, and microsystems of social support. Demographics, exposure to community violence (macrosystem), social support, social network (mesosystem), and perceived stress, mastery, and self-esteem (microsystem) were predictor variables. Depressive symptoms were measured by using the Center for Epidemiologic Studies of Depression (CES-D) Scale. FINDINGS: Variables from each system were significant predictors of depressive symptoms but perceived stress was the strongest predictor. Many postpartum adolescents reported that they had been victims of violence. Significant symptoms of depression were identified in 37% of the postpartum adolescents. CONCLUSIONS: Context is important to consider in comparing international studies of social support. Researchers and clinicians should investigate variables associated with the low incidence of treatment for depressive symptoms in postpartum adolescents. CLINICAL RELEVANCE: Feelings of high self-esteem and mastery should be fostered in nursing interventions with postpartum adolescents and routine screening for symptoms of depression should be considered in relevant healthcare settings. PMID- 18507566 TI - Maternal factors associated with child behavior. AB - BACKGROUND: Knowledge of the relative contributions of risk factors in predicting young children's behavior problems may provide insights for the development of preventive interventions. OBJECTIVE: The purpose of this cross-sectional study was to identify maternal predictors of children's internalizing and externalizing behaviors in a volunteer sample of 205 low-income, single mothers with children between 2 and 6 years of age. METHOD: Data were collected on chronic stressors, self-esteem, negative thinking, depressive symptoms, and child behavior during in home interviews with the mothers. RESULTS: Mothers' reports of internalizing and externalizing behaviors did not differ by sex or race of the child. Chronic stressors and depressive symptoms, in addition to control variables, explained 27% of the variability in internalizing behavior while these two variables accounted for 21% of the variability in externalizing behavior. For both internalizing and externalizing behavior, chronic stressors exerted the largest total effects. The effects of self-esteem and negative thinking were indirect, with the latter playing a stronger role. The indirect effect of negative thinking on child behavior was exerted through depressive symptoms, while self-esteem was linked with child behavior through both negative thinking and depressive symptoms. CONCLUSIONS: Decreasing mothers' negative thinking, a variable amenable to intervention, may not only decrease a mother's depressive symptoms but also improve her perception of the child's behavior. CLINICAL RELEVANCE: Decreasing mothers' negative thinking may provide a way to reduce their depressive symptoms and result in fewer behavior problems among their young children. Nurses working in primary care and community-based settings are in key positions to address this problem and improve the mental health of low-income mothers and positively affect the behavior of their children. PMID- 18507567 TI - Relative contribution of intrapersonal and partner factors to contraceptive behavior among Taiwanese female adolescents. AB - PURPOSE: To explore the relative contribution of intrapersonal factors (demographic data, sexual history, and self-efficacy for contraception) and partner factors (perceptions of support from sexual partners for contraception, and relationship power) to contraceptive behaviors among sexually active female adolescents in Taiwan. DESIGN: A cross-sectional design. Female adolescents who have had a steady male sexual partner in the past 3 months (N=375) were recruited as participants. METHODS: Questionnaires including demographic data, sexual history, contraceptive behavior, self-efficacy for contraception, perceptions of support from sexual partner for contraception, and perceptions of relationship power were submitted anonymously for this study. FINDINGS: Participants who had their first sexual experience at less than 14 years of age and were from one parent families had the least comprehensive contraceptive behavior than did other participants. Number of steady sexual partners was significantly negatively correlated with contraceptive behavior. Self-efficacy, perceptions of support from sexual partner for contraception, and relationship power all were positively correlated with contraceptive behavior. The important explanatory variables of contraceptive behavior were self-efficacy, age of first sexual intercourse, intervals between sexual intercourse, and perceptions of support from sexual partner for contraception. These accounted for 39.1% of variance in contraceptive behavior. CONCLUSIONS: Intrapersonal factors (self-efficacy, age of first sexual intercourse, and intervals between sexual intercourse) were more important than were partner factors (perceptions of support from sexual partners for contraception and relationship power) in influencing contraceptive behavior among sexually active female adolescents in Taiwan. CLINICAL RELEVANCE: Intervention to increase contraceptive behavior among female adolescents should be focused more on intrapersonal factors than on partner factors. PMID- 18507568 TI - Maternal worries, home safety behaviors, and perceived difficulties. AB - PURPOSE: The aim of the study was to explore the worries, safety behaviors, and perceived difficulties in keeping children safe at home in a purposive sample of low-income, predominantly non-English speaking mothers as a foundation for later nursing interventions. DESIGN: This study was a qualitative, descriptive design with content analysis to identify maternal concerns, behaviors, and perceptions of home safety as part of a larger study. METHODS: Eighty-two mothers, 64% of whom were monolingual Spanish-speakers, responded in writing to three semistructured interview questions. When mothers were unable to read and write the researcher wrote the responses, then read the content aloud for verification. A standardized probe for each question was posed to obtain richer responses. Data management included use of the software program NUD*IST and coding analyses following the Miles and Huberman guidelines (1994). Interpretations were translated into English for this report. FINDINGS: The major worries were falling, health, kidnapping, and being hit by a car. The leading maternal behaviors were coded as being physically, verbally, and environmentally preventive. Mothers said that it was their role to provide safety, and that this role could be wearisome, such that constant supervision was difficult. CONCLUSIONS: Low-income mothers described their worries for their 1 to 4 year-old children, explored their behaviors for preventing injury, and discussed what made keeping children from harm difficult. Understanding how mothers keep children safe, the barriers to home safety, and effective safety behaviors are important to the health of children. CLINICAL RELEVANCE: The clinical relevance of this study includes building trust as clinicians plan assessment, intervention and evaluation of home safety to encourage dialog about concerns, safety behaviors, and barriers to keeping children from injury. PMID- 18507569 TI - Evaluation of injury databases as a preliminary step to developing a triage decision rule. AB - PURPOSE: To evaluate accessibility and appropriateness of the Crash Outcomes Data Evaluation Systems (CODES) databases for prehospital trauma triage decision-rule development for people age 65 years and older. DESIGN AND METHODS: This informatics feasibility study included five steps for evaluating the accessibility of CODES databases. Eight criteria were used to evaluate the appropriateness of these databases for older person prehospital trauma triage decision-rule development. FINDINGS: Only 4 of the 33 states funded for CODES database development released their data to the study team during the 13-month data-acquisition period. Potential predictor variables (of life-threatening injury) and outcome variables (need for trauma center care) were identified for each database. Several databases had large amounts of missing data. Codebooks were available but descriptions of data validation procedures were unavailable. CONCLUSIONS: At this time, limited access to and development of CODES databases and missing data preclude the usefulness of these databases for older person triage decision rule development. Although adequate funding must be appropriated for continued CODES development, and commitment from individual states is essential, these databases offer great promise as a mechanism for decision-rule development to guide triage decision-making. Investigators should systematically evaluate large databases before using them in secondary analyses for clinical decision rule development. CLINICAL RELEVANCE: Nurses participate in the planning, development, and implementation of health information systems in various settings. They also assume important roles in prehospital care as direct care providers, EMS administrators, participants of local and state EMS councils where emergency care problems are discussed and policies are formulated, and through use at state and federal levels. PMID- 18507570 TI - Predictors of home healthcare nurse retention. AB - PURPOSE: To examine the level of job satisfaction and test a theoretical model of the direct and indirect effects of job satisfaction, and individual nurse and agency characteristics, on intent to stay and retention for home healthcare nurses. DESIGN: A descriptive correlation study of home healthcare nurses in six New England states. METHODS: Home healthcare nurse job satisfaction self-report data was collected with the HHNJS survey questionnaire & Retention Survey Questionnaire. FINDINGS: Based on a structural equation model, job tenure and job satisfaction were the strongest predictors of nurse retention. CONCLUSIONS: Understanding the variables associated with home healthcare nurse retention can help agencies retain nurses in a time of severe nurse shortages and increased patient demand. CLINICAL RELEVANCE: Predicted nursing shortages and increasing demand have made the retention of experienced, qualified nursing staff essential to assure access to high-quality home healthcare services in the future. PMID- 18507571 TI - The effect of high nursing surveillance on hospital cost. AB - PURPOSE: The purpose of this study was to determine the cost of one nursing treatment, surveillance, for older, hospitalized adults at risk for falling. DESIGN: An observational study using information from data repositories at one Midwestern tertiary hospital. The inclusion criteria included patients age>60 years, admitted to the hospital between July 1, 1998 and June 31, 2002, at risk for falls or received the nursing treatment of fall prevention. METHODS: Data came from clinical and administrative data repositories that included Nursing Interventions Classification (NIC). The nursing treatment of interest was surveillance and total hospital cost associated with surveillance was the dependent variable. Propensity-score analysis and generalized estimating equations (GEE) were used as methods to analyze the data. Independent variables related to patient characteristics, clinical conditions, nurse staffing, medical treatments, pharmaceutical treatments, and other nursing treatments were controlled for statistically. FINDINGS: The total median cost per hospitalization was $9,274 for this sample. The median cost was different (p=0.050) for patients who received high versus low surveillance. High surveillance delivery cost $191 more per hospitalization than did low surveillance delivery. CONCLUSION: Propensity scores were applied to determine the cost of surveillance among hospitalized adults at risk for falls in this observational study. The findings show the effect of high surveillance delivery on total hospital cost compared to low surveillance delivery and provides an example of a useful method of determining cost of nursing care rather than including it in the room rate. More studies are needed to determine the effects of nursing treatments on cost and other patient outcomes in order for nurses to provide cost-effective care. Propensity scores were a useful method for determining the effect of nursing surveillance on hospital cost in this observational study. CLINICAL RELEVANCE: The results of this study along with possible clinical benefits would indicate that frequent nursing surveillance is important and might support the need for additional nursing staff to deliver frequent surveillance. PMID- 18507572 TI - Health beliefs and mammography rates of Turkish women living in rural areas. AB - OBJECTIVES: Breast cancer is the most frequently occurring cancer among women globally as well as in Turkey and mammography is known to be an effective screening tool for this disease. This study was undertaken to characterize health beliefs and practices related to mammography of women in Turkey and also to survey mammography rates. DESIGN: Cross-sectional. The study was performed between September 2006 and January 2007 in Samsun, Turkey. Of 503 women, aged 35 years or older and registered at a local Health Center, 387 (76.94%) agreed to participate in this study. METHODS: A self-administered descriptive questionnaire and the Champion Revised Health Belief Model Scale for breast cancer screening (CHBMS) were used as data collection instruments. Analysis included descriptive statistics, Chi-square, and independent t test. Logistic regression analysis was conducted to identify the extent to which individual variables significantly predicted mammography use. RESULTS: Study participants did not undergo mammography at optimal rates (23.5%), but perceived benefits were higher than were perceived barriers to the procedure after education about mammography. Consequently women reported they were likely to adopt the practice of regular mammography in the future. CONCLUSIONS: Mammography rates were low in this rural area in Turkey, but may improve after education about the procedure. CLINICAL RELEVANCE: Mammography is an important tool in the early diagnosis of breast cancer. Many women still do not understand the benefit of mammograms and further education may help increase rates of early screening. PMID- 18507573 TI - The experiences of Western expatriate nursing educators teaching in Eastern Asia. AB - PURPOSE: To describe the lived experience of English-speaking Western nurse educators teaching in East Asian countries. DESIGN: The study design was an application of existential phenomenological approach to qualitative data collection and analysis. Eight expatriate nurse educators who had taught more than 2 years in an East Asian country were interviewed about their experiences between January 2004 and November 2005. METHODS: Narrative data were analyzed using a phenomenological approach aimed at distilling the experience of the educators. Each member of the research team analyzed the data, then through group discussions a consensus was reached, paying close attention to developing clear understandings of language nuances and maintaining the participants' voices. FINDINGS: Four themes emerged from the data: (a) differing expectations, (b) the cost of the expatriate experience, (c) bridging pedagogies, and (d) adapting and finding purpose. The tacit meanings of cultural differences affecting participants' experiences are presented. CONCLUSIONS: The ways that a collectivist-oriented culture may affect nurse educators coming from a Western individualist worldview needs to continue to be researched to develop better mutual understandings that will lead to culturally collaborative models of nursing practice, education and research. CLINICAL RELEVANCE: Nurses providing direct care and nurse educators who work with people from cultures other than their own will find the discussion of cross-cultural misunderstandings useful. PMID- 18507574 TI - The expatriate experience: teaching nursing across eastern and Western cultures. PMID- 18507575 TI - Advanced and specialist nursing practice: attitudes of nurses and physicians in Israel. AB - BACKGROUND: With the introduction of new and advanced nursing roles, the nursing profession is undergoing dynamic change. Realizing changes will be easier to accomplish if the nursing community and other healthcare professionals welcome the process. Recently the nursing staff mix in Israel has been undergoing a transformation: encouraging registered nurses to enhance their status by acquiring academic degrees and advanced professional training, and initiating the adoption of new nursing roles. OBJECTIVE: Our goal is to evaluate Israeli nurses' and physicians' attitudes to the introduction of new nursing roles and to expanding the scope of nursing practice. METHODS: Two hundred and fifteen nurses and 110 physicians from three large general hospitals and 15 community clinics filled in a questionnaire. FINDINGS: In general the majority of the nurses supported expansion of nursing practice, and such expansion did not cause significant opposition among physicians. However when the task affected patients' health, physicians were less willing to permit nurses to perform skills previously their responsibility alone. In addition, using multiple logistic regressions, support of the expansion of nursing practice was significantly higher among nurses in management or training positions, and among academically accredited nurses. Support for expanded roles was prominent among hospital physicians, graduates from Israeli schools of medicine, and less-tenured physicians. CONCLUSIONS: We suggest that confirmation by various groups of physicians and nurses of standardized definitions of the new boundaries in the scope of nursing practice roles could successfully promote development of new roles and facilitate integration of the Israeli healthcare system into the global context of change. CLINICAL RELEVANCE: Inter- and intra-professional collaboration, agreement, and understanding regarding advanced nursing practice roles and their introduction into the healthcare system might improve the relationship between healthcare professions and ultimately increase quality of care and patient satisfaction. PMID- 18507576 TI - Creating cross-disciplinary research alliances to advance nursing science. AB - PURPOSE: To demonstrate the utility of a developmental approach to building and sustaining cross-disciplinary research alliances. ORGANIZING CONSTRUCT: Cross disciplinary collaborations offer great promise for advancing an evolving program of research. Funding agencies' commitment to interdisciplinary work further heightens these collaborations. Nurse researchers, however, are not necessarily equipped with the knowledge and skills needed to strategically and successfully build and maintain cross-disciplinary collaborations. METHODS: Successful strategies for forging, extending, and sustaining alliances are organized using a developmental approach and illustrated by exemplars. FINDINGS: The initial phase of forging alliances is focused on creating a limited network to support relatively clear-cut research goals. The cohesive cross-disciplinary team is subsequently extended beyond the initial team to strategically address broader research goals. As the team matures, emphasis shifts toward sustaining the team through shared leadership that more actively focuses on the development of the field than the development of individual programs of research. CONCLUSIONS: The advancement of nursing science requires nurse researchers to rethink the developmental phases of building appropriate research teams. Strategic approaches for forging, extending, and sustaining cross-disciplinary alliances position nurse researchers as leaders in innovative cross-disciplinary research. The development of cross-disciplinary teams supports translational research and, by extension, the science that undergirds practice. CLINICAL RELEVANCE: Research that reflects cross-disciplinary perspectives is particularly well suited to addressing the complex health issues encountered by patients, families, and communities. PMID- 18507577 TI - Go to Ruth's house: a response to infant mortality. PMID- 18507578 TI - Go to Ruth's House. PMID- 18507579 TI - A randomized controlled trial of continuous labor support for middle-class couples: effect on cesarean delivery rates. AB - BACKGROUND: Previous randomized controlled studies in several different settings demonstrated the positive effects of continuous labor support by an experienced woman (doula) for low-income women laboring without the support of family members. The objective of this randomized controlled trial was to examine the perinatal effects of doula support for nulliparous middle-income women accompanied by a male partner during labor and delivery. METHODS: Nulliparous women in the third trimester of an uncomplicated pregnancy were enrolled at childbirth education classes in Cleveland, Ohio, from 1988 through 1992. Of the 686 prenatal women recruited, 420 met enrollment criteria and completed the intervention. For the 224 women randomly assigned to the experimental group, a doula arrived shortly after hospital admission and remained throughout labor and delivery. Doula support included close physical proximity, touch, and eye contact with the laboring woman, and teaching, reassurance, and encouragement of the woman and her male partner. RESULTS: The doula group had a significantly lower cesarean delivery rate than the control group (13.4% vs 25.0%, p = 0.002), and fewer women in the doula group received epidural analgesia (64.7% vs 76.0%, p = 0.008). Among women with induced labor, those supported by a doula had a lower rate of cesarean delivery than those in the control group (12.5% vs 58.8%, p = 0.007). On questionnaires the day after delivery, 100 percent of couples with doula support rated their experience with the doula positively. CONCLUSIONS: For middle-class women laboring with the support of their male partner, the continuous presence of a doula during labor significantly decreased the likelihood of cesarean delivery and reduced the need for epidural analgesia. Women and their male partners were unequivocal in their positive opinions about laboring with the support of a doula. PMID- 18507580 TI - Tears in the vagina, perineum, sphincter ani, and rectum and first sexual intercourse after childbirth: a nationwide follow-up. AB - BACKGROUND: The first sexual intercourse after childbirth may be challenging for women, especially if the birth resulted in injuries in the genital area. The purpose of this study was to investigate whether or not tears in the vagina, perineum, sphincter ani, or rectum hindered sexual intercourse during the year after childbirth. METHODS: We obtained information from 2,490 women in a population-based cohort identified at antenatal care. Information about first sexual intercourse was collected by means of a questionnaire sent 1 year after birth to the women and about women's tears reported in the population-based Swedish Medical Birth Register. RESULTS: Adjusted relative risks with 95 percent confidence intervals for not having had sexual intercourse within 3 and 6 months, respectively, after childbirth were 1.5 (95% CI 1.2-1.8) and 1.6 (95% CI 1.2-2.3) for tears in the vagina, 1.4 (95% CI 1.1-1.6) and 1.5 (95% CI 1.1-2.1) for tears in the perineum, and 2.1 (95% CI 1.4-3.1) and 2.2 (95% CI 1.1-4.6) for tears in the sphincter ani and rectum. No statistically significant differences were found at 1-year follow-up. No associations between episiotomy and delay in resuming intercourse were found after adjusting the relative risks. CONCLUSIONS: Tears in the vagina, perineum, sphincter ani, or rectum are associated with a delay in women's resumption of sexual intercourse 6 months after childbirth in Sweden. PMID- 18507581 TI - Perinatal factors related to negative or positive recall of birth experience in women 3 years postpartum in the Netherlands. AB - BACKGROUND: Little research has been conducted to date on women's postnatal emotional well-being and satisfaction with the care received in the Netherlands. The aim of this study was to investigate Dutch women's views of their birth experience 3 years after the event. METHODS: A questionnaire was mailed to all women who had given birth in 2001 and who had at least one prenatal, perinatal, or postnatal visit to the participating midwifery practice. Women who had a subsequent birth after the index birth in 2001 were not excluded. We specifically asked respondents to reflect on the birth that occurred in 2001. Women were asked to say how they felt now looking back on their labor and birth, with five response options from "very happy" to "very unhappy." RESULTS: We received 1,309 postnatal questionnaires (response rate 44%). The sample was fairly representative with respect to the mode of delivery, place of birth, and obstetric interventions compared with the total Dutch population of pregnant women; however, the sample was not representative for ethnicity and initial caregiver. Three years after delivery, most women looked back positively on their birth experience, but more than 16 percent looked back negatively. More than 1 in 5 primiparas looked back negatively compared with 1 in 9 multiparas. Adjusted odds ratios (OR) for looking back negatively 3 years later included having had an assisted vaginal delivery or unplanned cesarean delivery (OR 2.6, 95% CI 1.59 4.14), no home birth (OR 1.4, 95% CI 1.04-1.93), referral during labor (OR 2.4, 95% CI 1.48-3.77), not having had a choice in pain relief (OR 2.9, 95% CI 1.91 4.45), not being satisfied in coping with pain (OR 4.9, 95% CI 2.55-9.40), a negative description of the caregivers (OR 2.9, 95% CI 1.85-4.40), or having had fear for the baby's life or her own life (OR 2.3, 95% CI 1.47-3.48). CONCLUSIONS: A substantial proportion of Dutch women looked back negatively on their birth experience 3 years postpartum. Further research needs to be undertaken to understand women's expectations and experiences of birth within the Dutch maternity system and an examination of maternity care changes designed to reduce or modify controllable factors that are associated with negative recall. PMID- 18507582 TI - Cesarean section: Norwegian women do as obstetricians do--not as obstetricians say. AB - BACKGROUND: The worrying trend of an ever-increasing incidence of delivery by cesarean section has been commented on repeatedly. Studies from the United Kingdom and the United States have found that many obstetricians would choose cesarean section for themselves without strict medical indication, whereas similar studies from Denmark and Norway have indicated that almost none would choose cesarean section for themselves. The purpose of this study was to report the proportion of Norwegian obstetricians who have children born by cesarean section and to compare the rate with that among other physicians and that with the general population. METHODS: Questionnaires were sent to 1,500 random members of the Norwegian general public, 1,500 randomly selected physicians, and 423 random surgeons asking whether they had children born by cesarean section. All were between the ages of 40 and 65 years. RESULTS: The response rate was 78 percent. In the general public with children, 12 percent reported that one or more of them were born by cesarean section. The average was 8 percent among those with only basic schooling compared with 16 percent (p < 0.02) among those who had been to university for more than 4 years. This figure was 19 percent among physicians in general (p < 0.001 compared with the general population), 26 percent among surgeons, and 27 percent among the 189 specialists in obstetrics and gynecology (p < 0.02 compared with the physicians in general). CONCLUSION: The rate of cesarean section in the general population is unlikely to fall as long as so many obstetricians have their own children delivered by cesarean section. PMID- 18507583 TI - Impact of first childbirth on changes in women's preference for mode of delivery: follow-up of a longitudinal observational study. AB - BACKGROUND: A woman's childbirth experience has an influence on her future preferred mode of delivery. This study aimed to identify determinants for women who changed from preferring a planned vaginal birth to an elective cesarean section after their first childbirth. METHODS: This prospective longitudinal observational study involved two units that provide obstetric care in Hong Kong. A mail survey was sent to 259 women 6 months after their first childbirth. These women had participated in a longitudinal cohort study that examined their preference for elective cesarean section in the antenatal period of their first pregnancies. Univariate and multivariate analyses were performed to identify determinants for women who changed from preferring vaginal birth to elective cesarean section. RESULTS: Twenty-four percent (23.8%, 95% CI 18.4-29.3) of women changed from preferring vaginal birth to elective cesarean section after their first childbirth. Determinants found to be positively associated with this change included actual delivery by elective cesarean section (OR 106.3, 95% CI 14.7 767.4) intrauterine growth restriction (OR 19.5, 95% CI 1.1-353.6), actual delivery by emergency cesarean section (OR 8.4, 95% CI 3.4-20.6), higher family income (OR 3.2, 95% CI 1.1-8.8), use of epidural analgesia (OR 2.6, 95% CI 1.0 6.8), and higher trait anxiety score (OR 1.1, 95% CI 1.0-1.3). The most important reason for women who changed from preferring vaginal birth to elective cesarean section was fear of vaginal birth (24.4%). CONCLUSIONS: A significant proportion of women changed their preferred mode of delivery after their first childbirth. Apart from reducing the number of cesarean sections in nulliparous women, prompt provision of education to women who had complications and investigations into fear factors during vaginal birth might help in reducing women's wish to change to elective cesarean section. PMID- 18507584 TI - Obstetricians' and midwives' attitudes toward cesarean section. AB - BACKGROUND: The cesarean section rate has increased worldwide over the past 20 years; in Italy, it is now more than 35 percent. Although clinical factors are important, the attitudes of health practitioners toward cesarean section need further investigation to correctly identify facilitators and barriers to changes. The objective of this study was to explore the attitudes toward cesarean section of midwives and obstetricians who worked in the same geographical area. METHODS: Face-to-face structured interviews using an adaptation of the Survey of Clinicians' Views on Caesarean Section, an anonymous questionnaire with 35 open and closed answers on practitioners' views on cesarean section, were conducted. The questionnaire was given to the entire group of midwives and obstetricians working in Modena, a northern Italian district. RESULTS: Of 262 eligible practitioners, 248 were interviewed (response rate 94.6%). The midwives' attitudes toward cesarean section differed from those of the obstetricians. Sixty five percent of midwives considered the rates of cesarean section in their hospitals to be too high compared with 34 percent of obstetricians (p < 0.001). Midwives were also less inclined to believe that cesarean section provides benefits to the mother (p = 0.02) or that it is indicated by previous cesarean delivery (p < 0.001). No differences were observed between male and female obstetricians. CONCLUSIONS: In this survey, the attitudes toward cesarean section were correlated more with professional role than with gender. This information can help policy makers to shape interventions aimed at providing better care for pregnant and childbearing women. PMID- 18507585 TI - Economic implications of home births and birth centers: a structured review. AB - BACKGROUND: It is widely perceived that home births and birth centers may help decrease the costs of maternity care for women with uncomplicated pregnancies and deliveries. This structured review examines the literature relating to the economic implications of home births and birth center care compared with hospital maternity care. METHODS: The bibliographic databases MEDLINE (from 1950), CINAHL (from 1982), EMBASE (from 1980), and an "in-house" database, Econ2, were searched for relevant English language publications using MeSH and free text terms. Data were extracted with respect to the study design, inclusion criteria, clinical and cost results, and details of what was included in the cost calculations. RESULTS: Eleven studies were included from the United Kingdom, United States, Australia, and Canada. Two studies focused on home births versus other forms and locations of care, whereas nine focused on birth centers versus other forms and locations of care. Resource use was generally lower for women cared for at home and in birth centers due to lower rates of intervention, shorter lengths of stay, or both. However, this fact did not always translate into lower costs because, in the U.K. where many studies were conducted, more midwives of a higher grade were employed to manage the birth centers than are usually employed in maternity units, and because of costs of converting existing facilities into delivery rooms. The quality of much of the literature was poor, although no studies were excluded for this reason. Selection bias was likely to be a problem in those studies not based on randomized controlled trials because, even where birth center eligibility was applied throughout, women who choose to deliver at home or in a birth center are likely to be different in terms of expectations and approach from women choosing to deliver in hospital. CONCLUSIONS: This review highlights the paucity of economic literature relating to home births and birth centers. Differences in results between studies may be attributed to differences in health care systems, differences in methods used, and differences in costs included. Further economic research that involves detailed bottom-up costing of alternative options for place of birth and measures multiple outcomes, including women's preferences, would help address the question of whether out-of-hospital birth is beneficial in economic terms. PMID- 18507586 TI - Do women with epilepsy have more fear of childbirth during pregnancy compared with women without epilepsy? A case-control study. AB - BACKGROUND: Although anxiety and depression in populations with epilepsy have been studied, no research on fear of childbirth in women with epilepsy have been conducted. The purposes of this study were to examine whether a significant difference occurred in fear of childbirth between pregnant women with epilepsy and pregnant healthy controls and to evaluate the most common fears. METHODS: Fifty pregnant women with epilepsy and 50 pregnant women without epilepsy were assessed at a gestational age between 32 and 36 weeks of pregnancy, using two questionnaires for the measurement of fear of childbirth, an open question, and a clinical interview. RESULTS: We found that during pregnancy, women with epilepsy experienced a significantly higher rate of fear of childbirth when compared with healthy controls, whereas after delivery, the experiences and feelings about childbirth are almost the same as those of women without epilepsy. CONCLUSIONS: Our findings signify the importance of the assessment of the fear of childbirth, especially in women with epilepsy, and the need to offer professional and ad hoc support to those who suffer from it. PMID- 18507587 TI - Events after stillbirth in relation to maternal depressive symptoms: a brief report. AB - BACKGROUND: Actions taken after a stillbirth can affect long-term psychological morbidity. Our objective was to study how infant bonding and maternal actions after stillbirth are associated with ensuing depressive symptoms. METHODS: Using the population-based Swedish Medical Birth Register, we identified all 380 Swedish-speaking women who gave birth to singleton stillborn infants in Sweden in 1991. Of these, 314 (83%) completed a postal questionnaire 3 years after the stillbirth. Items included actions taken to bond with the baby and demographics. The association between care-related factors and later maternal depressive symptoms was quantified using relative risks estimated using multivariable regression. RESULTS: We observed an almost sevenfold increased risk of depressive symptoms for mothers who reported not being with their babies as long as they wished (adjusted risk ratio [RR] 6.9, 95% CI 2.4-19.8). Compared with women who became pregnant again within 6 months, those with no later pregnancy were at higher risk of depressive symptoms (adjusted RR 2.8, 95% CI 0.9-8.4). In addition, compared with women who experienced a stillbirth in their first pregnancy, stillbirth occurring with an infant who was third in the birth order was related to a twofold risk of elevated depressive symptoms (adjusted RR 2.2, 95% CI 0.8-6.4). Furthermore, stillbirth occurring in a fourth or later pregnancy was associated with an almost sevenfold risk of depressive symptomatology (adjusted RR 6.7, 95% CI 2.2-20.5). No evidence of an association was found between other care-related actions and subsequent maternal depressive symptoms. CONCLUSIONS: Our results suggest that a mother being with the stillborn baby for as long as desired and the birth order of the stillbirth may influence her later depressive symptomatology. Compared with mothers who became pregnant again within 6 months, those who did not have a subsequent pregnancy were at higher risk of depressive symptoms at 3 years' follow-up. PMID- 18507588 TI - Marginalization of midwives in the United States: new responses to an old story. AB - This column addresses issues raised by an intensive study of the circumstances and actions that resulted in the closure of two long-standing, successful nurse midwifery services in a large United States city in 2003. Dr. Steffie Goodman of the School of Nursing, University of Colorado Health Science Center in Denver, USA, conducted 52 in-depth interviews with midwives, nurses, administrators, childbirth educators, policymakers, and physicians in an effort to understand how and why these two services were closed and what their closures revealed about the general underutilization of midwives in contemporary U.S. health care. Goodman concluded that economics, power, and authority converge in a way that allows persons in positions of institutional power and authority to make self-serving decisions that diminish access to midwifery services and that they can do so without any public accountability for their actions. PMID- 18507592 TI - Warm compresses in second stage did not lower the rate of sutured perineums in primiparous women. PMID- 18507593 TI - Keeping the editor "in focus". PMID- 18507594 TI - On health care reform. PMID- 18507596 TI - Computer-generated versus nurse-determined strategy for incubator humidity and time to regain birthweight. AB - OBJECTIVE: To compare effects on premature infants' weight gain of a computer generated and a nurse-determined incubator humidity strategy. An optimal humidity protocol is thought to reduce time to regain birthweight. DESIGN: Prospective randomized controlled design. SETTING: Level IIIC neonatal intensive care unit in the Netherlands. PARTICIPANTS: Infants of 24 to 30 weeks gestational age with a birthweight less than 1,500 g. INTERVENTION: Two incubator humidity strategies were studied: computer-generated and nurse-determined humidity. MAIN OUTCOME MEASURE: Time needed to regain birthweight. RESULTS: One hundred thirty six infants were enrolled: 65 were exposed to the computer-generated strategy and 71 to the nurse-determined strategy. Demographic characteristics were well balanced between groups, with birthweight 981 +/- 245 versus 991 +/- 213 g, mean gestational age 27.7 +/- 1.7 versus 27.7 +/- 1.6 weeks. Main outcome did not significantly differ between strategies: survival analysis showed an equal number of days needed to regain birthweight (median 9 days, with 95% CIs 8-10 and 7-11 for infants exposed to the computer-generated and nurse-determined humidity strategy, respectively). CONCLUSION: Computer-generated strategy does not reduce the time needed to regain birthweight. PMID- 18507597 TI - Predictors of wheezing in prematurely born children. AB - OBJECTIVE: To examine the degree to which neonatal illness severity, postneonatal health problems, child characteristics, parenting quality as measured by the HOME Inventory, and maternal characteristics are related to the development of wheezing in prematurely born children over the first 27 months after term. DESIGN: Longitudinal predictive study. SETTING: Infants were recruited from two neonatal intensive care units, one in southeast and one in Midwest. PARTICIPANTS: One hundred thirteen preterm infants who weighed less than 1,500 g or required mechanical ventilation and their mothers. MAIN OUTCOME MEASURES: The presence of wheezing was obtained from maternal report at 2, 6, 9, 13, 18, 22, and 27 months. Wheezing was considered to be medically significant if the child was using bronchodilators or pulmonary antiinflammatory medications. RESULTS: Sixty-eight percent of the children had wheezing at least one or more ages; 47% of the children were also taking bronchodilators or pulmonary antiinflammatory medications and thus had medically significant wheezing. CONCLUSION: Postneonatal health problems and the social environment appear to be more important in developing wheezing in prematurely born children than neonatal medical complications. PMID- 18507598 TI - Maternal bedsharing practices, experiences, and awareness of risks. AB - OBJECTIVES: The objectives of this study were to determine mothers' practices and experiences of bedsharing with their infants and also to determine their knowledge of the risks. DESIGN: A self-report questionnaire was mailed to 1,122 mothers of infants. SETTING: Manitoba, Canada. PARTICIPANTS: The participants in this study were the mothers of infants aged 3 months whose contact information was obtained through the Manitoba Health Information Privacy Committee. MAIN OUTCOME MEASURE: Maternal-infant bedsharing practices and maternal knowledge of risks. RESULTS: There were 293 completed questionnaires received (26% return rate). Key findings suggest that although 89% of participants agreed that sleeping with one's baby has some risks associated with it, 72% reported that they bedshared with their baby on either a regular or an occasional basis. Mothers who breastfed were twice as likely to have bedshared. Approximately 13% of respondents who had bedshared reported an experience(s) with bedsharing in which they had rolled onto or partway onto their infant. CONCLUSIONS: Although the majority of participants agreed that bedsharing had risks for infants, almost three quarters of respondents reported bedsharing on a regular or an occasional basis. Reports by respondents of rolling onto or partway onto their infants support the conclusion that health care professionals should promote safe sleeping environments that include a separate sleep surface for infants. PMID- 18507599 TI - Relationship of genital injuries and age in adolescent and young adult rape survivors. AB - OBJECTIVE: To examine the associations between age and genital injuries in adolescent and young adult women examined following rape. DESIGN & SETTING: A retrospective review of 234 medical records from an emergency department sexual assault program. SAMPLE: Women aged 14 to 29 years. Fifty percent of the sample was African American, 48% was White, and 2% was either Asian or an "other" race. MAIN OUTCOME MEASURES: Genital injury was described by injury prevalence, frequency, and anatomical locations of injuries. RESULTS: Overall genital injury prevalence was 62.8%. Younger age was not significantly associated with the presence or absence of genital injury. However, younger age was significantly associated with an increased number of genital injuries overall and to the thighs, labia minora, periurethral area, fossa navicularis, and vagina. CONCLUSION: These findings support the need for further research to determine if the current care provided to rape survivors is age appropriate. PMID- 18507600 TI - Perinatal beliefs and practices of immigrant Punjabi women living in Canada. AB - OBJECTIVE: To describe new immigrant Punjabi women's perinatal experiences and the ways that traditional beliefs and practices are legitimized and incorporated into the Canadian health care context. DESIGN: Naturalistic qualitative descriptive. PARTICIPANTS/SETTING: Fifteen first-time mothers who had immigrated in the past 5 years to Canada from Punjab, India, and had given birth to a healthy infant in the past 3 months in a large urban center in British Columbia, Canada. Five health professionals and community leaders also took part in a focus group to confirm the study findings and to offer recommendations. RESULTS: Three major categories emerged: the pervasiveness of traditional health beliefs and practices related to the perinatal period (e.g., diet, lifestyle, and rituals), the important role of family members in supporting women during the perinatal experiences, and the positive and negative interactions women had with health professionals in the Canadian health care system. CONCLUSIONS: Change is required at the levels of the health professional, the heath care system, and the community to ensure that culturally safe care is provided to immigrant Punjabi women and their families during the perinatal period, which is an important and sensitive period of interaction with the Canadian health care system. PMID- 18507601 TI - A systematic review of telephone support for women during pregnancy and the early postpartum period. AB - OBJECTIVE: To assess the effects of telephone-based support on smoking, preterm birth, low birthweight, breastfeeding, and postpartum depression. DATA SOURCES: Cochrane Pregnancy and Childbirth Group trials register (March 2006), Cochrane Central Register of Controlled Trials (March 2006), Medline (1966-2006), EMBASE (1980-2006), and CINAHL (1982-2006). Secondary references were scanned and experts in the field were contacted. STUDY SELECTION: All published, unpublished, and ongoing randomized controlled trials of telephone support interventions in which the primary aim was smoking, preterm birth, low birthweight, breastfeeding, or postpartum depression were reviewed. DATA EXTRACTION: Data were independently extracted by both authors and double entered into the Cochrane Collaboration's Review Manager (2003) software. DATA SYNTHESIS: Trials evaluating different primary outcomes were analyzed separately. For dichotomous data, results were presented as summary relative risk with 95% confidence intervals. For continuous data, weighted mean difference was used. CONCLUSIONS: Proactive telephone support may (a) assist in preventing smoking relapse, (b) play a role in preventing low birthweight, (c) increase breastfeeding duration and exclusivity, and (d) decrease postpartum depressive symptomatology. No telephone interventions were effective in improving preterm birth or smoking cessation rates. Additional research is encouraged. PMID- 18507602 TI - A literature update on maternal-fetal attachment. AB - OBJECTIVE: To critically review and synthesize original research published since 2000 designed to measure factors that influence maternal-fetal attachment. DATA SOURCES: EBSCOhost Research Databases that included PubMed, CINAHL Plus, PsycINFO, and SCOPUS were searched for journal articles published in the past 7 years (2000-2007) that examined variables thought to increase, decrease, or cause no change in level of maternal-fetal attachment. Keyword searches included maternal-fetal attachment, parental attachment, and prenatal attachment. STUDY SELECTION: Twenty-two studies were selected that met the inclusion criteria of original research, clear delineation of the measurement of maternal-fetal attachment, measurement of maternal-fetal attachment during pregnancy, and inclusion of women or couples, or both. DATA EXTRACTION: Studies measuring maternal-fetal attachment included a broad range of variables as potential risk or protective factors, or both. Factors associated with higher levels of maternal fetal attachment included family support, greater psychological well-being, and having an ultrasound performed. Factors such as depression, substance abuse, and higher anxiety levels were associated with lower levels of maternal-fetal attachment. DATA SYNTHESIS: The large majority of studies reviewed were limited by small, homogenous samples deemed insufficient to detect significant differences, inconsistent measurement of maternal-fetal attachment during gestational periods, and cross-sectional designs. CONCLUSIONS: Further research is essential to identify factors influencing maternal-fetal attachment. Specifically, research needs to be conducted on larger sample sizes of greater racial and ethnic diversity. PMID- 18507603 TI - A review of the evidence comparing the human papillomavirus vaccine versus condoms in the prevention of human papillomavirus infections. AB - OBJECTIVE: To examine the evidence related to the efficacy of condom use versus the human papillomavirus vaccine in the prevention of human papillomavirus infections. DATA SOURCES: Cochrane, CINHAL, PubMed, and Clinical Evidence. Various combinations of the keywords HPV, vaccine, and condoms were used for the search. STUDY SELECTION: Randomized, double-blinded placebo-controlled trials were reviewed for evaluation of the human papillomavirus vaccine. Several longitudinal studies and a meta-analysis were used for review of condom efficacy related to human papillomavirus transmission. DATA EXTRACTION AND SYNTHESIS: Studies evaluating the use of either condoms or the human papillomavirus vaccine and its impact on human papillomavirus transmission rates, detected through either human papillomavirus DNA testing or clinical disease. CONCLUSIONS: The evidence indicates that the greatest degree of protection from specific types of human papillomavirus infection is provided by the vaccine. However, the use of condoms in addition to the human papillomavirus vaccine provides the greatest protection from the untoward effects of human papillomavirus infection and may also provide protection against human papillomavirus types not in the vaccine and other sexually transmitted infections. PMID- 18507604 TI - A review of psychometric properties of feeding assessment tools used in neonates. AB - OBJECTIVE: To appraise the psychometric properties of clinical feeding assessment tools used in a neonatal population. DATA SOURCES: PubMed, OvidMedline, CINHAL, and PsycINFO databases from 1980 to 2007. Reference lists of all identified articles were also reviewed. STUDY SELECTION: Research reports written in English that utilized or validated clinical feeding assessment tools. DATA EXTRACTION: In total, 941 articles were reviewed. Seven neonatal clinical feeding assessment tools were identified and categorized into three groups: tools used for assessing either bottle-feeding or breastfeeding behaviors, tools used only for assessing bottle-feeding behaviors, and tools used only for assessing breastfeeding behaviors. RESULTS: The psychometric properties of none of the seven assessment tools identified were satisfactory, and the limited representativeness of the samples of the psychometric research was noted in all assessment tools identified. The Neonatal Oral-Motor Assessment Scale appeared to have been examined more thoroughly and showed more consistent results in psychometric properties than the others, despite its own limitations. CONCLUSIONS: A psychometrically sound neonate feeding assessment tool has not yet been empirically validated. Clinicians who use these tools for clinical and research purposes should take into account this lack of evidence of psychometric soundness and interpret results of assessment with precautions. Well-designed research is needed to study the scientific integrity of these instruments for program evaluations in neonatal care. PMID- 18507605 TI - Annotated bibliography of NINR findings on women's health in pregnancy and childbirth: 2008 update. PMID- 18507606 TI - If Margaret Sanger could see us now. PMID- 18507608 TI - The hysteroscopic approach to sterilization. AB - Permanent sterilization is the most common form of contraception in the United States today. Previously, this was accomplished via the laparoscopic method. A new less invasive method has been developed, in which the Essure microinsert is inserted through a hysteroscope into the fallopian tubes. A subsequent benign inflammatory process results in tubal occlusion within 3 months of insertion. Approved since 2002, this method offers high efficacy without incisions, general anesthesia, or a prolonged recovery period. PMID- 18507609 TI - Implanon: a new contraceptive implant. AB - Women who cannot use hormonal contraception containing estrogen have a variety of progestin-only contraceptive methods from which to choose. Implanon is a new single-rod progestin-only contraceptive implant that is easily inserted and can remain in place for up to 3 years. It is highly effective with a rapid onset of action and an equally rapid return of fertility once removed. Counseling is important to help women decide if this method is appropriate for their needs. PMID- 18507610 TI - The latest advances in hormonal contraception. AB - Millions of women each year start or continue to use some type of hormonal contraceptive method. Choosing a method may be anxiety provoking. In this article, we review some of the latest advances in and options for hormonal contraception, including extended-dose oral contraceptives, the vaginal ring, injectable methods, and emergency contraception. Nurses can facilitate women's decision making for healthy reproductive options. PMID- 18507611 TI - Options for intrauterine contraception. AB - The IUD is a convenient and effective contraceptive option for many women. Currently, there are two different types of intrauterine contraception available to women in North America: the levonorgestrel-releasing intrauterine system and the copper T IUD. A greater understanding of the benefits and limitations of these two contraceptive options will assist women's health care providers to better meet the family planning needs of their patients. PMID- 18507612 TI - New options for barrier contraception. AB - Barrier contraceptives are a safe alternative to hormonal methods of fertility management. Newer barrier method options include the Today Sponge, the FemCap, and the Lea's Shield. Understanding the use, benefits, and limitations of these barrier methods of birth control will assist women's health care providers to better meet the family planning needs of their patients. PMID- 18507613 TI - Abstracts of the XXVII Congress of the European Academy of Allergology and Clinical Immunology (EAACI). Barcelona, Spain. June 7-11, 2008. PMID- 18507614 TI - Anatomy of the ansa cervicalis: nerve fiber analysis. AB - Ansa cervicalis complex, which innervates the infrahyoid muscles, is formed by the union of two roots derived from the anterior primary rami of the first three or four cervical spinal nerves. According to its relationship with the internal jugular vein, there are three ansa types: medial, lateral, or mixed. Nerve fiber microscopy was carried out in 18 ansa cervicalis complexes after staining with Sudan Black, in order to study composition and structural arrangement in detail. Fibers from both first and second cervical nerves (C1, C2) joined the hypoglossal nerve and later left it partially in the superior root of the ansa, which also contained ascending fibers derived from the inferior root components; usually the second and third cervical nerves. Those ascending fibers, together with the C1 and C2 fibers remaining in the hypoglossal nerve, innervate thyrohyoid and geniohyoid muscles. Cervical nerve fibers could be observed within the hypoglossal nerve even after the ramification of those branches. Further, cervical nerve fibers were continuously issued to the peripheral layers of the hypoglossal nerve where the fibers of the two nerves intermingled, forming a complex structure. The general arrangement of the ansa did not change according to its type or segmental composition. Although the hypoglossal nerve does not make any significant contribution to the formation of the ansa, the close relationship between the two nerves observed in the present study calls for the use of a more descriptive term such as ansa hypoglosso-cervicalis. The pattern of innervation of the infrahyoid muscles is also discussed. PMID- 18507615 TI - Staging of disuse atrophy of skeletal muscles on immunofluorescence microscopy. AB - The Japanese population is rapidly aging, thereby causing excess demand for facilities for elderly invalids. It is imperative that social measures and scientific studies be carried out to enable better care of bedridden elderly people. The purpose of the present study was to review the histological changes that occur in disuse atrophy of skeletal muscles, the primary pathophysiology of bedridden invalids, with the object of developing a staging standard to be used by researchers and clinicians. Rat hindlimb suspension was used as an experimental model. Atrophy of the soleus muscle was evaluated qualitatively and quantitatively on immunofluorescence microscopy. The myofibrils decreased significantly in the first 2-3 weeks of disuse atrophy. The earliest morphological change was fan-shaped multistep forking of sarcomeres, which appeared by the first week. This type of muscular lesion, designated here as 'sarcomeric disarray', was first described in the present study. Central-core lesions appeared mainly in slow muscle fibers by the second week. These lesions disappeared by the fourth or fifth week. Nerves remained intact and no inflammation or regeneration occurred up to the fifth week. Methods and criteria were compiled for staging of disuse atrophy based on the present results and a diagnosis kit designed for studies on disuse atrophy of skeletal muscles. PMID- 18507616 TI - Zygomaticofacial, zygomaticoorbital and zygomaticotemporal foramina: anatomical study. AB - The zygomatic nerve may be disrupted when elevating periorbita from the lateral wall during orbital surgery, and care should be taken to prevent injury to this nerve during lateral orbitotomy approaches to access intraorbital soft-tissue tumors. Furthermore, the precise introduction of anesthetics to the zygomaticoorbital (ZO), zygomaticofacial (ZF) and zygomaticotemporal (ZT) foramina could be important data for the plastic and reconstructive surgeon operating in the area. The aim of the present study was to investigate the morphologic and topographic anatomy, and variations of the ZO, ZF and ZT. The present study was performed using 200 dry human skulls. The ZF, ZO and ZT foramina varied from being absent to as many as four small openings. We classified each of these foramina as types I-V for single, double, triple, quadruple and absent foramina, respectively. The relative frequency was as follows: type I, ZO 50%, ZF 40%, ZT 30%; type II, ZO 20%, ZF 15%, ZT 15%; type III, ZO 10%, ZF 5%, ZT 5%; type IV, ZO 3%, ZF 1%, ZT-; and type V, ZO 17%, ZF 39%, ZT 50%. A detailed knowledge of the anatomic morphometry of this area is necessary for a surgeon when performing maxillofacial surgery and regional block anesthesia. Anatomic variations in this area may be present and a surgeon must take this into consideration so as to increase surgical success. PMID- 18507617 TI - Alternative approach to evaluating lumbar lordosis on direct roentgenograms: projection area per length squared. AB - Several methods have been described to evaluate the degree of lumbar lordosis. However, suggested methods have used non-standardized terminology and landmarks to measure the degree of lumbar lordosis. In the present study a practical method for evaluating the degree of lumbar lordosis is described and, for this purpose, 24 lateral roentgenograms were obtained retrospectively from the archive of Department of Physical Medicine and Rehabilitation, Ondokuz Mayis University, Samsun, Turkey. The length between the superior and inferior angles of the first and fifth lumbar vertebral bodies, and the area behind the lumbar vertebral bodies, were estimated using the point counting and planimetry methods. A new unit, the projection area per length squared (PAL) was described on lateral roentgenograms. The planimetric approach was used as the gold standard in the present study. The point-counting method was also used to estimate the PAL and it was repeated three times to determine the variability of the technique. To evaluate the estimates' accuracy, the results of point-counting were compared with those of the planimetry methods. The PAL changed by between 3.93 and 13.59% for the examined subjects. A high correlation was also noted between the results of the point-counting and planimetry methods (r = 0.997). It is concluded that the PAL approach could provide accurate and reproducible data for evaluating the degree of lumbar lordosis and low back pain. PMID- 18507618 TI - Morphological and functional changes of the rat parotid glandular cells by clipping and reopening the parotid duct, using HAM8 antibody. AB - The purpose of this experiment is to examine the proliferative process of rat acinar cells after parotid duct ligation and reopening. Two experimental groups were observed. The first group was killed from 0 to 14 days after the duct ligation. In the second group, the duct was clipped for 14 days, and it was reopened. Following a period of from 2 to 28 days after removal of the clip, the glands were removed to perform a histological analysis, including hematoxylin eosin (HE), immunofluorescent staining using HAM8 antibody, which recognizes connexin 32, and transmission electron microscopy (TEM). In the experimental gland from the 1st group at 6 days after ligation (I-6D), the acinar cells disappeared. In the tissue from the 2nd group 8 days after reopening (II-8D), newly formed acinar cells were found again. Lobular structure of the parotid glands recovered in the II-21D. HAM8 signals were observed between normal acinar cells, while they declined in the tissue from I-1D, and they were not observed in the I-2D. HAM8 signals were first observed in the II-25D and then subsequently returned to normal levels in the II-28D. These results suggest that the intercellular communication and functional recovery was not complete 25 days after reopening of the duct.In conclusion, the recovery of the acinar structure was recognized during an extended period of duct ligation, however, a time lag between the morphological and functional recovery was found to exist. PMID- 18507619 TI - Variations in the configuration of the circle of Willis. AB - Brains were obtained from 1000 medicolegal autopsy subjects of varying ages. The circle of Wills was examined at the base of the brain. The cerebral vessels were observed with regards to their origin, caliber and typical configuration. Variations were noted and grouped into different categories. Out of 1000 specimens examined, 452 (45.2%) conformed to the typical pattern. In the rest of the specimens (54.8%) there were variations in the circulus arteriosus. The circle was deficient in 32 (3.2%). The anterior cerebral artery was absent in 0.4%; hypoplastic in 1.7%; duplicated in 2.6%; triple in 2.3% and single in 0.9%. The anterior communicating artery was absent in 1.8%, duplicate in 10%, triplicate in 1.2% and plexiform in 0.4%. Multiplication of posterior cerebral artery was observed in 2.4% cases while it was hypoplastic in 10.6% brains. Posterior communicating artery was absent in 1% and hypoplastic in 13.2%. Seventy four brains (7.4%) had multiple variations. Intracranial saccular aneurysm was present in 10 (1%). These figures are compared with the available literature. Persistence of some embryonic vessel that normally disappear, disappearance of vessels that would normally persist or sprouting of new vessels due to hemodynamic and genetic factors are the usual causes for such anomalies. These variations are discussed with regard to development and other hemodynamic factors. PMID- 18507620 TI - Four-headed biceps and triceps brachii muscles, with neurovascular variation. AB - Anatomical variations of the biceps brachii and triceps brachii have been described by various authors, but the occurrence of four-headed biceps brachii and triceps brachii in an ipsilateral arm is rare and has not been reported before in the literature. During routine cadaveric dissection in the department of anatomy, Kasturba Medical College, Mangalore, India, various unusual variants were noted in the left arm of a cadaver of a 67-year-old man. The variants include a four-headed biceps, a four-headed triceps, communication between the musculocutaneous and median nerves, and a high origin of deep brachial artery from the axillary artery. The occurrence of these anomalies and the clinical and morphological significance are discussed. PMID- 18507621 TI - Nerve communication between the glossopharyngeal nerve, external carotid plexus and the superficial cervical ansa: human autopsy case. AB - The authors encountered a very rare human autopsy case in which the supernumerary branch of the glossopharyngeal nerve and a nerve branch arising from the external carotid plexus communicated with the superficial cervical ansa. This anomaly was observed on the left side of a 71-year-old male cadaver during the gross anatomical seminar at Niigata University in 2004. The nerve fascicle and fiber analyses indicated that the supernumerary branch of the glossopharyngeal nerve separated cranial to the branches to the pharyngeal constrictor muscles, carotid sinus and stylopharyngeal muscle and sent the nerve fibers to the muscular branches to the platysma and the cutaneous branches to the cervical region. Additionally, it was shown that the branch arising from the external carotid plexus sent the nerve fibers to the cutaneous branch to the cervical region. Although the external carotid plexus is primarily postganglionic sympathetic fibers originating from the superior cervical ganglion, the vagus and glossopharyngeal nerves gave off branches connecting to the plexus, and therefore it was not possible to determine the origins of this branch of the external carotid plexus. The present nerve fascicle analysis demonstrates that the supernumerary branch of the glossopharyngeal nerve, which innervated the platysma, did not share any nerve components with the branches to the pharyngeal constrictor muscles, carotid sinus and stylopharyngeal muscle, suggesting that this supernumerary branch may be categorized into the different group from these well-known branches. PMID- 18507622 TI - Supratrochlear foramen of the humerus. PMID- 18507624 TI - Abstracts of the Ninth Congress of the European Society of Contact Dermatitis. May 28-31, 2008. Estoril, Portugal. PMID- 18507625 TI - Abstracts of the 18th Annual Meeting of the European Childhood Obesity Group. June 5-7, 2008. Porto, Portugal. PMID- 18507626 TI - Abstracts of the 34th European Congress of Cytopathology. June 15-18, 2008. Rovaniemi, Finland. PMID- 18507627 TI - Cold pearl surfactant-based blends. AB - Pearlizing agents have been used for many years in cosmetic formulations to add a pearlescent effect. Cold pearl surfactant-based blends are mixtures of glycol stearates and surfactants which can be blended in the cold into a wide range of personal-care formulations to create a pearlescent lustre effect. Under controlled manufacturing conditions constant viscosities and crystalline characteristics can be obtained. The development of these blends has been driven by efforts to improve the economics of adding solid pearlizing agents directly into a hot mix formulation. This paper summarizes the history of pearlizers, describes their advantages and physical chemistry of the manufacturing process. Finally some suggestions for applications are given. Les agents nacrants sont utilises depuis de nombreuses annees dans les formulations cosmetiques pour ajouter un effet nacre. Les melanges a froid a base de tensioactif nacre sont des melanges de stearates de glycol et de tensioactifs qui peuvent etre melanges a froid dans une large gamme de formulations d'hygiene personnelle pour creer un effet de lustre nacre. On peut obtenir des viscosites et des proprietes cristallines constantes avec des conditions de fabrication maitrisees. Le developpement de ces melanges a ete porte par les efforts pour ameliorer les couts de l'ajout d'agents nacrants solides directement dans une formulation melangee de l'ajout d'agents nacrants solides directement dans une formulation melangee a chaud. Cet article resume l'histoire des agents nacrants, decrit leurs avantages et al physico-chimie du procede de fabrication. On emet a la fin cetaines suggestions d'applications. PMID- 18507628 TI - The use of radiolabelling techniques to measure substantivity to, and penetration into, hair of protein hydrolysates. AB - The use of 14C-labelled amino acids enables the measurement of both the total substantivity to hair and the degree of penetration into the hair shaft of amino acid mixtures derived from complete hydrolysis of proteins. The technique utilizes the fact that direct measurement of 14C radioactivity of the treated hair detects only the surface substantivity. Total substantivity can be determined following solubilization of the hair. Data obtained for wheat amino acids show significant penetration when used to treat hair from a shampoo or conditioner formulation. A similar technique has been investigated for a wheat protein partial hydrolysate using 14CNO for radiolabelling purposes and shows that significant penetration into hair can occur. L'utilisation d'amino-acides marques au 14C permet la mesure a la fois de l'absorption totale par les cheveux et du degre de penetration dans la fibre du cheveu de melanges d'amino-acides obtenus a partir d'une hydrolyse totale de proteines. La technique utilise le fait que la mesure directe de la radioactivite du 14C des cheveux traites ne detecte que l'absorption en surface. L'absorption totale peut etre determinee apres solubilisation des cheveux. Les donnees obtenues a partir d'amino-acides de ble montrent une penetration significative lors d'une utilisation pour traiter des cheveux a partir d'une formulation de shampoing ou d'apres-shampoing. Une technique similaire a ete exploree vis-a-vis d'un hydrolysat partiel d'une proteine de ble utilisant 14CNO a des fins de marquage, et montre qu'une penetration significative dans les cheveux peut avoir lieu. PMID- 18507629 TI - Environmentally compatible surfactants for the cosmetic industry. AB - From the application pattern of surfactant-containing cosmetic products, it is inevitable that the major part of the chemicals will be discharged into waste water and eventually will enter the environment. Because the environmental compatibility of the products is based on the ecological properties of their raw meterials, the biodegradability and ecotoxicological behaviour of the product components and particularly the surfactants, form the central elements of the environmental compatibility assessment. The tools for this evaluation are standardized test systems, which are described and discussed on the basis of the ecological data of selected surfactants. De par le type d'application des produits cosmetiques contenant des tensioactifs, il est inevitable que la plus grande partie des substances chimiques soit evacuee dans les eaux usees et finisse par arriver dans l'environnement. Puisque la compatibilite environnementale des produits est basee sur les proprietes ecologiques de leurs matieres premieres, la biodegradabilite et le comportement ecotoxicologique des composants des produits, et en particulier des tensioactifs, forment les elements majeurs de l'evaluation de la compatibilite environnementale. Les outils de cette evaluation sont des systemes d'essai normalises, qui sont decrits et commentes d'apres les donnees ecologiques de tensioactifs choisis. PMID- 18507630 TI - Amphoteric surfactants: processing, product composition and properties. AB - Cocamidopropyl betaine (CAPB) has been the most important secondary surfactant for personal-cleansing products for a long time. Its excellent toxicological profile is an important reason for its increasing use in oral-care products. Recently it has gained interest for further applications such as household cleaners, dish-washing liquids, and industrial and technical products. Imidazoline-derived amphoterics such as sodium cocoampho-acetate (SCAA) or diacetate play a more minor role than CAPB. Owing to the low irritation potential of the pure surfactant and its good toxicological properties, ampho-acetates have mainly found applications in cosmetics. Their industrial applications have been relatively small. While CAPB has a well-defined chemical structure from a straightforward production process, most imidazoline-derived amphoterics exhibit a complex composition of compounds with different structures. This depends on the production parameters. Improved processing methods have recently led to the commercial availability of well-defined SCAA with low levels of by-products. Modern production processes and the composition of high-purity amphoterics are reviewed. Raw materials and by-products are described, together with their analytical methods. The cosmetic performance, cleansing and foaming power, rheological effects and mildness-enhancing properties of both CAPB and SCAA are compared. La cocamidopropyl-betaine (CAPB) est, depuis longtemps le tensio-actif secondaire le plus important pour les produits d'hygiene personnelle. L'excellent profil toxicologique de la CAPB est certainement une raison majeure de son usage croissant dans les produits de soin buccaux. La CAPB a suscite depuis peu un interet pour des applications supplementaires telles que les nettoyants menagers, les liquides vaisselle, les produits industriels et techniques. Les derives amphoteres de l'imidazoline tels que le cocoampho-acetate de sodium (SCAA) ou le diacetate occupent une place mineure comparee a la CAPB. En raison du faible potentiel irritant du tensio-actif pur et meme de bonne proprietes toxicologiques, les ampho-acetates trouvent principalement leurs applications dans les cosmetiqes. Leur role dans les applications industrielles est relativement limite. Alors que la CAPB a une structure chimique bien definie a partir d'un procede de production direct, la plupart des derives amphoteres d'imidazoline presentent une composition complexe de composes aux structures differentes. Ceci depend des parametres de production. Des procedes de production ameliores ont recemment conduit a une disponibilite commerciale de SCAA bien definis avec de faibles teneurs en produits secondaires. Les procedes modernes de production et la composition d'amphoteres de grande purete sont decrits. Les matieres premieres et les produits secondaires sont decrits ainsi que leurs methodes d'analyse. Le comportement cosmetique, le pouvoir nettoyant et moussant, les effets rheologiques et les proprietes adoucissantes sont compares, a la fois pour la CAPB et le SCAA. PMID- 18507631 TI - Erratum. PMID- 18507632 TI - Influence of topically applied ceramide/phospholipid mixture on the barrier function of intact skin, atopic skin and experimentally induced barrier damage. AB - The influence of a liposomal formulation of a ceramide/phospholipid mixture on the barrier function of the skin in atopic tests, in subjects with healthy skin, and after lipid extraction was tested. The transepidermal water loss and the stratum corneum hydration served as parameters for the evaluation of the influence of the ceramide/phospholipid-mixture treatment on the barrier function. Additionally a barrier-function test with nicotinic acid ester was performed after ceramide/phospholipid-mixture application. Stabilizing effects on the barrier function after application of ceramide/phospholipid-mixture were found only in atopic skin and after lipid extraction but not in healthy skin. The results lead us to expect positive effects of ceramide/phospholipid-mixture only if barrier damage is present. PMID- 18507633 TI - The use of block copolymer stabilizers for controlling dispersion stability. AB - The role of block copolymers in stabilizing colloidal dispersions against flocculation or coalescence is discussed in terms of the detailed structure of the interface. Both particulate dispersions and liquid/liquid interfaces are used as examples. Two novel experimental methods are introduced, small-angle neutron scattering and neutron reflection. Comparison with the Scheutjens Fleer theory for block copolymers is also presented. Strategies for the optimum design of the stabilizing polymer are introduced. PMID- 18507634 TI - Zinc and titanium oxides: promising UV-absorbers but what influence do they have on the intact skin? AB - Suspensions (20%) of microfine and ACS grade zinc oxide, or 20% titanium oxide in water, castor oil or polyethylene glycol were applied topically to rabbit skin over 4 h (1 day) or 2 h daily (3 days). Skin sites were analysed for metal uptake. Percutaneous absorption of the oxides was probably low, most remaining on the skin surface. Uptake patterns were influenced by the vehicles used. Appreciably more microfine zinc oxide was taken up by the skin in subacute studies. Local changes in trace metal concentration were construed as evidence of percutaneous absorption of zinc and titanium oxides. Further studies are indicated to validate the significance of these preliminary observations. PMID- 18507635 TI - Rheology of siloxane-stabilized water in silicone emulsions. AB - Using silicone copolymers in personal care products can improve the aesthetic performance of formulations. During their manufacture, distribution and topical application they are subject to various mechanical stresses. In this study rheology was used to measure their effects. A number of water in silicone (w/Si) emulsions were prepared in which the oil phase consisted of cyclomethicone. The surfactant used was a branched type silicone copolymer. Both viscoelastic and viscometry measurement were performed on model systems and on commercial products. Experimental data were obtained using a Bohlin rheometer. The measurements were taken applying shear rates in the range of 0.46-58 l s-1 and for the strain sweep frequencies of 0.1 Hz, 1 Hz and 10 Hz were applied. Oscillation tests were performed in the 0.1 Hz to 10 Hz range. All measurements were taken at 35deg;C, representing the approximate temperature encountered during topical application. The effect of surfactant concentration on viscoelastic properties was examined. It was shown that with increasing surfactant concentration the elastic moduli G' and the viscous moduli G" increased. Furthermore, the emulsions showed a transition from a predominantly elastic to a predominantly viscous response as the surfactant concentration increased. The effect of varying the water phase volume fraction on viscometry and viscoelastic measurements was also examined. With increasing water phase volume fraction the viscosity of the emulsions, as well as the yield stress, increased. The Cross and Sisko models were applied. From the Dougherty and Krieger equation phieff was calculated. It was found the the data derived from the Sisko model gave more reliable results. Results obtained from commercial samples showed a high proportion of elasticity; oscillation tests and viscometry experiments suggested that tumbling had the biggest impact on the theological profiles; viscosity, eta, shear stress, sigma, elastic module, G', and dynamic viscosity, eta', dropped to a minimum in these samples. Results from the two commercial samples were compared and it was observed that, although both were w/o emulsions, different rheological behaviour could be observed. PMID- 18507636 TI - Design and in vivo evaluation of ultrafine inorganic-oxide-containing-sunscreen formulations. AB - Ultrafine titanium dioxide, zinc oxide, and a 1:1 mixture of ZnO/TiO2 were used as 5% dispersions in sunscreen formulae. Three different carrier bases were evaluated for their accelerated stability and rheological properties with and without metal oxides based on sodium lauryl sulphate/polysorbate 80, triethanolamine stearate, and cetyl trimethyl ammonium chloride. All three bases showed thixotropic behaviour. Addition of metal oxides only affected the magnitude of viscosity not the viscoelastic behaviour. The cationic emulsion base was found to be the most stable to incorporate the microfine metal oxides, the others' viscosity values showing a significant drop in storage. The in vivo sun protection factors were determined on the cationic based emulsions in four human subjects using an UV source covering both UVA-UVB regions. The mean sun protection factors were found to be 5.03, 4.03, and 4.8 for TiO2, ZnO, and 1:1 ZnO/TiO2 respectively for 4 mg.cm-2 applications, the differences not being significant. PMID- 18507637 TI - Skin smoothing effects of Dead Sea minerals: comparative profilometric evaluation of skin surface. AB - The skin smoothing effects of three different liquid gels were compared on 20 mature women. Treatment applications were performed twice a day over a period of 4 weeks, and the skin roughness parameter (Rz) of all test participants was determined at the beginning and at the end of the study using a computer-aided laser profilometry, in accordance with DIN 4768 ff. At the end of the application period, the liquid gel with 1% of a Dead Sea mineral solution had an average skin roughness parameter reduction of 40.7%. The liquid gel without mineral additives showed an average reduction in skin roughness of 27.8%. The control gel without anti-wrinkle agents or the additives showed an average reduction of only 10.4%. PMID- 18507638 TI - Prolonged effects of antidandruff shampoos - time to recurrence of Malassezia ovalis colonization of skin. AB - Most active antidandruff shampoos exhibit a strong activity against the yeast Malassezia ovalis. The present study was undertaken to compare the prolonged antifungal effect of three proprietary shampoos containing either 2% ketoconazole, 1.5% zinc pyrithione or 2.5% selenium sulphide. Superficial squames were harvested from the scalp in the days following a 6-week antifungal shampoo treatment. Counts of yeasts highlighted by a fluorochrome were made using computerized image analysis. Data show the increased duration of yeast reduction for the ketoconazole shampoo over the two other formulations. The lingering effect of ketoconazole is explained by the combination of its antifungal and pharmacokinetic properties. PMID- 18507639 TI - Characterization of the mineral content of a physical sunscreen emulsion and its distribution onto human stratum corneum. AB - When using physical sunscreens, protection and aesthetic aspects are directly related to particle size and mineral distribution onto skin. These investigations aimed to characterize the different mineral raw materials entering into the composition of a specific sunscreen emulsion and to assess the distribution of the sunscreeen agents at the surface of human stratum corneum. X-ray diffractometry revealed the crystallographic features of the TiO2 and ZnO crystals. Frequency distributions of crystal length and aspect ratio were determined for the zinc oxide which presented the largest particle size and varying shapes. Scanning and transmission electron microscopy disclosed structural information. The results reported in this study specify the characteristics of the oxide particles involved in the formulation and show a satisfactory mineral distribution along the irregular topography of the skin. No intracellular penetration was noted. PMID- 18507640 TI - A model for the shine of hair arrays. PMID- 18507641 TI - Application of high-performance liquid chromatography in the analysis of direct dyes in semipermanent hair colouring cosmetics. AB - A method for the identification and the quantification of direct dyes in semipermanent hair colouring cosmetics by using a reversed phase high-performance liquid chromatography (HPLC) with gradient elution and diode array detector (DAD) detection is presented. A standard mixture of 18 commonly marketed dyestuffs (11 nitroderivatives, two anthraquinones and five Arianor dyes) was studied. The major problems in the analysis of the mixture of direct dyes is the different chemical structure and especially the wide range of polarities. However we succeeded in the complete separation of the standard dyes. A qualitative analysis of eight colouring commercial products selected from different shades and a quantitative analysis of three of them was carried out. Materials, methods and results are reported. PMID- 18507642 TI - Occlusive power evaluation of O/W/O multiple emulsions on gelatin support cells. AB - The water content of the stratum corneum plays an important role in providing skin suppleness and smoothness. The diffusion of water through the skin is limited primarily by the stratum corneum and the noncornified part of the epidermis has negligible water barrier properties. Multiple emulsions are vesicular systems utilized for the prolonged liberation of active ingredients. The O/W/O multiple emulsion type is employed in cosmetics because its high oil content is able to maintain an occlusive film (barrier) on the skin surface. The objective of this study was to determine the occlusive power of O/W/O multiple emulsions on gelatin support cells. The results showed that occlusive products form a uniform layer on the surface of gelatin after the test, whereas nonocclusive products form two layers: an aqueous phase on the gelatin, and an oil phase above the aqueous phase. Thus, the different occlusive powers are due to the homogeneity of this layer and to its ability to prevent water evaporation. PMID- 18507643 TI - Vapour pressure of some fragrance ingredients in emulsion and microemulsion formulations. AB - The vapour pressure of two fragrance raw materials, phenethyl alcohol and phenethyl acetate, in a microemulsion system is discussed against their participation in the different phases of the system. The results are used to evaluate the change of the fragrance vapour pressure during water evaporation from a typical emulsion system. PMID- 18507644 TI - Sunscreens' photochemical behaviour: in vivo evaluation by the stripping method. AB - This study shows the influence of a water-in-oil emulsion and an oil-in-water emulsion on the photochemical behaviour of four sunscreen preparations containing various amounts of three UV filters: octyl methoxycinnamate, butyl methoxydibenzoylmethane, and 4-methylbenzylidene camphor. With the stripping method in vivo, skin-product interactions were evaluated after topical application and ultra-violet exposure. Evaluations were also made under the same operating conditions as in determining sun protection factor in vivo. Results showed that each sunscreen preparation has a particular photochemical behaviour related to filter mixture and emulsion type. When developing a sunscreen preparation the choice of the emulsion type and filters is relevant for achieving optimum efficacy and spectral stability. PMID- 18507645 TI - FTIR investigation of the damage produced on human hair by weathering and bleaching processes: implementation of different sampling techniques and data processing. AB - Untreated and bleached hair samples were analysed by FTIR spectroscopy. Cysteic acid and cysteine-S-thiosulphate were analysed by different data manipulations and various sampling techniques. Single-fibre analysis was achieved using the IR microscope attachment to the FTIR instrument (transmission mode). An ATR diamond crystal was also used for both single fibres and tresses; this sampling technique was compared to a contact sample ATR cell based on a ZnSe crystal. The microscope was found to be very useful for the analysis of small portions of hair fibres; the effect of weathering along the length of the fibres and the extent of variability among the single fibres was assessed by this technique. The results clearly show a trend in the cysteic acid content increasing from the mid-point to the tip; it was found that hair samples are very heterogeneous. A standard variation of up to 11% is reported for the mean value of the intensity of absorption of cysteic acid in untreated fibres belonging to the same tress sample. The value of the variation decreases when the tress is bleached -- the longer the bleaching process the lower the value of the standard variation (4.5% for hair fibres bleached for 60 min using hydrogen peroxide). The contact sample ATR cell gave poor reproducibility as only poor sample-crystal contact was achieved and the level of noise recorded was too high to allow accurate detection. However the ATR diamond crystal was found to give sensitive and reproducible spectra since much higher levels of transmittance were achieved (better sample-crystal contact); this technique allowed measurements of groups of seven to nine hair fibres at once, but did not provide reproducible spectra of single fibres. Different data manipulations have been assessed; these include derivatization and calculation of peak heights and areas (ratioed by the intensity of absorption of the peaks chosen as internal standards to the amide III 1232--1242 cm-1 and CH2 stretching 1451 cm-1), normalization, averaging spectra and spectral subtraction. PMID- 18507646 TI - The effect of a synthetic ceramide-2 on transepidermal water loss after stripping or sodium lauryl sulfate treatment: an in vivo study. AB - The integrity of the chemical and physical structure of the horny layer is essential for maintaining the skin in good health. Any disturbance of this integrity may lead to cutaneous reactions of varying degree: dryness, redness, inflammation. The measurement of Transepidermal Water Loss (TEWL) allows one to record this kind of disturbance and to follow the slow return to normal. In this in vivo study two techniques of insulting the epidermis were used: stripping and washing with sodium lauryl sulfate (SLS). A significant increase of TEWL values resulted in both cases. The application of emulsions containing 0.5% and 1% of a synthetic ceramide type-2 (N-stearoyl-DL-erythro-sphinganine) decreased the disturbance measured by TEWL, in a significant fashion in both trials. The placebo emulsions showed no significant effect. The ceramide thus seemed to participate in the restructuring of the horny layer. PMID- 18507647 TI - Utilization of electrochemical silver ions as preservative agent in cosmetic dispersions. AB - The use of anodic silver ions as preserving agents in cosmetics was tested by a challenge test in a set of cosmetic dispersions with the addition of known preservative inhibitors or micro-organism growth-promoters such as humectants, hydrosoluble collagen and vegetable extracts. Silver's microbicidal efficacy, compared to that of imidazolidinyl urea or methyl p-hydroxybenzoate, showed a more efficient activity especially in the presence of proteinaceous material. This agent may represent a good and safe protection for finished products both in manufacture and during use. PMID- 18507648 TI - Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo. AB - This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration. PMID- 18507649 TI - Sputum neurokinin A in Egyptian asthmatic children and adolescents: relation to exacerbation severity. AB - BACKGROUND: Neurogenic inflammation may participate in the development and progression of bronchial asthma. The molecular mechanisms underlying neurogenic inflammation are orchestrated by a large number of neuropeptides including tachykinins such as neurokinin A (NKA) and substance P. Tachykinins are secreted from sensory airway nerves and inflammatory cells after allergens exposure. In clinical practice, assessment of airway inflammation is difficult. Therefore, detection of biological markers of airway inflammation in sputum might offer help for proper monitoring of asthma severity. AIM OF THE STUDY: We aimed to measure sputum NKA in relation to acute asthma exacerbations of varying severity. METHODS: Sputum NKA was measured by enzyme-linked immunosorbent assay in 24 children and adolescents during and after acute asthma exacerbation and 24 healthy matched controls. RESULTS: Sputum NKA was significantly higher in asthmatic patients during acute exacerbation than controls [217.5 (284) vs 10 (7) ng/ml, P < 0.001]. When patients with acute asthma exacerbation were followed-up till remission, sputum NKA levels decreased significantly, but they remained significantly higher than controls. Sputum NKA levels were significantly higher in severe than moderate and in moderate than mild exacerbations, and was negatively correlated to peak expiratory flow rate (r = -0.9, P < 0.001). Sputum NKA had significant positive correlations to eosinophil counts in blood and sputum (r = 0.6, P < 0.001 and r = 0.7, P < 0.001 respectively). CONCLUSIONS: Sputum NKA is up-regulated during acute asthma exacerbation and it positively correlates to its severity. Thus, NKA may aid in objective classification of the exacerbation severity. In addition, NKA may be a target for new asthma therapy. PMID- 18507650 TI - A specific mixture of short-chain galacto-oligosaccharides and long-chain fructo oligosaccharides induces a beneficial immunoglobulin profile in infants at high risk for allergy. AB - BACKGROUND: It has been suggested that human breast milk oligosaccharides play a role in the development of the immune system in infants, and may consequently inhibit the onset of allergy. A specific prebiotic mixture of short-chain galacto oligosaccharides and long-chain fructo-oligosaccharides (GOS/FOS) has been shown to reduce the incidence of atopic dermatitis (AD) at 6 months of age in infants at risk for allergy. AIM OF THE STUDY: This study was aimed to analyze the effect of GOS/FOS on the immune response in these infants. METHODS: In a double-blind randomized placebo-controlled study, infants received a hypoallergenic whey formula with either 8 g/l GOS/FOS in a 9 : 1 ratio (IMMUNOFORTIS) or 8 g/l maltodextrine (placebo) for 6 months. At 3 months of age, children were vaccinated with Hexavac against a.o. diphteria, tetanus, polio (DTP). At 6 months of age, plasma samples were collected from 84 infants (verum group n = 41, placebo group n = 43). Levels of total immunoglobulins (Ig) and of cow's milk protein (CMP-) and DTP-specific Ig were measured. RESULTS: GOS/FOS supplementation led to a significant reduction in the plasma level of total IgE, IgG1, IgG2 and IgG3, whereas no effect on IgG4 was observed. CMP-specific IgG1 was significantly decreased. DTP-specific Ig levels were not affected. CONCLUSIONS: This study shows that GOS/FOS supplementation induces a beneficial antibody profile. GOS/FOS reduces the total Ig response and modulates the immune response towards CMP, while leaving the response to vaccination intact. This suggests that oral GOS/FOS supplementation is a safe method to restrain the atopic march. PMID- 18507651 TI - Linkage disequilibrium structures in cattle and their application to breed identification testing. AB - We examined the extent of linkage disequilibrium (LD) block lengths in four breed populations: Japanese Black, Angus, Hereford and Holstein. Three chromosomal regions in which QTL were previously mapped in Japanese Black populations were scanned with 84 microsatellite markers. The estimated LD lengths in these four purebred populations varied from 535 to 683 kb, which is much shorter than the values reported previously. Our findings suggest that QTL can be mapped in sub centimorgan regions in these populations using an LD-mapping method. We also developed breed identification methods to distinguish Japanese Black from Angus, Hereford, Holstein and F(1) animals (Japanese Black x Holstein) respectively using the haplotypic frequencies of a pair of markers in the breed populations. After assessing the distributions of posterior probabilities to be Japanese Black, we obtained several pairs of markers that completely distinguished Japanese Black from the other breeds. We also obtained several combinations of six markers that completely distinguished Japanese Black animals from F(1) animals. PMID- 18507652 TI - Exact trait-model-free tests for linkage detection in pedigrees. AB - A number of trait-model-free tests have been proposed for linkage detection between a genomic region and a trait. These tests involve testing the dependence in segregation between a trait and marker alleles by assigning a score to every possible identity-by-descent configuration of the pedigree members without modeling the trait, and then averaging the scores over all such configurations compatible with the observed marker genotypes and genealogical relationship of the pedigree members. In this paper we propose a permutation test as an alternative to the existing exact trait-model-free tests for linkage detection. The proposed test is computationally efficient and is applicable on complex multigeneration pedigree structures. In this paper, we have compared the performance of the permutation test with two other exact trait-model-free tests for linkage detection on simulated datasets. We have demonstrated that the proposed permutation test is fully robust against mispecification of marker allele frequencies and has very good power for linkage detection. The permutation test is implemented in the program lm_ibdtests within the framework of MORGAN 2.8 (http://www.stat.washington.edu/thompson/Genepi/MORGAN/Morgan.shtml). PMID- 18507653 TI - Alpha glucosidase inhibitor voglibose can prevent pioglitazone-induced body weight gain in Type 2 diabetic patients. PMID- 18507654 TI - Dose-effect study of domperidone as a galactagogue in preterm mothers with insufficient milk supply, and its transfer into milk. AB - AIMS: To investigate the possibility of a dose-response relationship for the use of domperidone in treating insufficient milk supply in mothers of preterm infants, and to quantify the exposure of the breastfed infant to domperidone. METHODS: Six preterm mothers received domperidone (30 mg daily or 60 mg daily) in a double-blind, randomized, crossover trial. Milk production and serum prolactin were measured before and during the trial, and domperidone concentration in milk was measured during drug treatment. RESULTS: For milk production, two of the mothers were 'nonresponders', whereas the other four were 'responders' and showed a significant increase in milk production from 8.7 +/- 3.1 g h(-1) in the run-in phase (mean +/- SEM), 23.6 +/- 3.9 g h(-1) for the 30-mg dose (P = 0.0217) and 29.4 +/- 6.6 g h(-1) for the 60-mg dose (P = 0.0047). In all participants, serum prolactin was significantly increased for both doses, but the response was not dose dependent. Median (interquartile range) domperidone concentrations in milk over a dose interval at steady-state were 0.28 microg l(-1) (0.24-0.43) and 0.49 microg l(-1) (0.33-0.72) for the 30-mg and 60-mg doses, respectively. The mean relative infant dose was 0.012% at 30 mg daily and 0.009% at 60 mg daily. CONCLUSION: In one-third of mothers, domperidone did not increase milk production. In the remainder, milk production increased at both domperidone doses, and there was a trend for a dose-response relationship. The amount of domperidone that transfers into milk was extremely low, and infant exposure via breastfeeding was not considered to be significant. PMID- 18507655 TI - Spironolactone use and the risk of upper gastrointestinal bleeding: a population based case-control study. AB - AIMS: Recent studies have suggested an increased risk of upper gastrointestinal bleeding (UGB) in spironolactone users. The aim was to confirm the association, identify the risk factors and quantify the absolute risk. METHODS: A population based case-control study was conducted in the County of Funen, Denmark. Cases (n = 3652) were all subjects with a first discharge diagnosis of serious UGB during the period 1995 to 2006. Age- and gender-matched controls (10 for each case) (n = 36 502) were selected by risk set sampling. Data on all subjects' drug exposure and past medical history were retrieved from a prescription database and from the County's patient register. Confounders were controlled by conditional logistic regression. RESULTS: The adjusted odds ratio (OR) associating current use of spironolactone with UGB was 2.7 [95% confidence interval (CI) 2.2, 3.2]. The risk increased with higher doses of spironolactone (5.4; 3.4, 8.6) for 100-mg tablets. No trend was found with increasing cumulative dose. The strongest association was found among users aged 55-74 years (OR 13.1; 6.5, 26.3). Current use of loop diuretics was also associated with an increased risk of UGB (1.9; 1.7, 2.1). CONCLUSION: The use of spironolactone is associated with increased risk of UGB. The risk increases with higher doses. PMID- 18507656 TI - The ability of atropine to prevent and reverse the negative chronotropic effect of fingolimod in healthy subjects. AB - AIMS: The authors determined whether intravenous atropine can prevent or counteract the negative chronotropic effect of the immunomodulator fingolimod. METHODS: In this randomized, placebo-controlled, two-period, crossover study, 12 healthy subjects received 5 mg fingolimod orally concurrently with intravenous atropine (titrated to a heart rate of 110-120 beats min(-1)) or intravenous placebo. A second group of 12 subjects received atropine/placebo 4 h after the fingolimod dose. Continuous telemetry measurements were made for 24 h after each fingolimod dose. RESULTS: Fingolimod administration alone yielded a heart rate nadir of 51 +/- 5 beats min(-1) at a median 4 h postdose with heart rate remaining depressed at 51-64 beats min(-1) over the rest of the day. Concurrent administration of fingolimod and atropine yielded a nadir of 66 +/- 6 beats min( 1) resulting in an atropine: placebo ratio (90% confidence interval) of 1.30 (1.22, 1.36). When atropine was administered at the time of the nadir, it was able to reverse the negative chronotropic effect of fingolimod from a heart rate of 56 +/- 9 beats min(-1) (placebo) to 64 +/- 8 beats min(-1) (atropine) resulting in an atropine: placebo ratio of 1.15 (1.04, 1.26). Atropine had no influence on the pharmacokinetics of fingolimod. CONCLUSIONS: Atropine administered concurrently with fingolimod prevented the heart rate nadir that typically occurs 4 h postdose. Atropine administered at the time of the heart rate nadir was able to reverse the negative chronotropic effect of fingolimod. PMID- 18507657 TI - Decompensation of chronic heart failure associated with pregabalin in a 73-year old patient with postherpetic neuralgia: a case report. PMID- 18507658 TI - The reproducibility of adenosine monophosphate bronchial challenges in mild, steroid-naive asthmatics. AB - AIMS: Repeated adenosine monophosphate (AMP) challenges are used to assess drug efficacy in clinical trials of mild, steroid-naive asthmatics. Refractoriness has been reported after repeated challenges over short intervals. This study evaluated possible tachyphylaxis after repeated AMP challenges at 12 and 24 h in mild, steroid-naive asthmatics. METHODS: This was an open, three-way crossover study. Twenty-six steroid-naive asthmatic subjects were randomized to the following AMP challenge regimens separated by 7-14 days: (A) challenge at 08.00 h, repeated 24 h later; (B) challenge at 08.00 h, repeated 12 and 24 h later; (C) challenge at 20.00 h, repeated 12 h later. Comparisons within day were assessed using 90% confidence intervals (CIs). Non-inferiority approach taken with 1 doubling concentration (DC) as a clinically relevant difference. RESULTS: Regimen A: Significant increase in AMP reactivity at 24 h. Mean DC difference was 0.6 (90% CI 0.24, 0.96). Regimen B: No evidence of difference between AMP reactivity at 08.00 h and a repeated challenge 12 h later. Repeated challenge at 24 h caused a significant increase in provocation concentration (PC)(20) compared with 12 h (mean DC difference 0.48, 90% CI 0.02, 0.95) and 0 h (mean DC difference 0.82, 90% CI 0.49, 1.14 - the upper CI exceeds the criteria of 1 DC). Challenge regimen C: No difference between challenges; mean DC difference of 0.28 (90% CI -0.2, 0.76). CONCLUSION: The small decline in AMP reactivity during repeated challenges was not consistently observed, and was small compared with the known effects of inhaled drugs. PMID- 18507659 TI - Melatonin as an adjuvant therapy in patients with rheumatoid arthritis. PMID- 18507660 TI - A signal of increased risk of hypoglycaemia with angiotensin receptor blockers caused by confounding. AB - AIMS: To study reporting of hypoglycaemia in angiotensin receptor blocker (ARB) users, and to investigate the possibility of confounding. METHODS: The French pharmacovigilance database was examined for an association between hypoglycaemia and ARBs or other drugs using reports notified between 1996 and 2005. This association was also tested in patients taking or not taking antidiabetic agents (ADAs) using reporting odds ratios (ROR). RESULTS: Hypoglycaemia was mentioned in 807 of the 174 595 reports entered during the study period. Overall hypoglycaemia was associated with the use of ARBs [ROR 2, 95% confidence interval (CI) 1, 3] and with the use of ADAs (ROR 32, 95% CI 27, 37). Moreover, the use of ARBs was associated with the use of ADAs (OR 7, 95% CI 6, 8). Considering separately reports with and without ADA, the association of ARB use with a higher risk of hypoglycaemia disappeared (OR 0.4, 95% CI 0.2, 0.8 and OR 2, 95% CI 1, 3, respectively). CONCLUSION: A signal indicating an association between ARB use and hypoglycaemia was found in the French pharmacovigilance database. This signal disappeared after stratification on ADA use, thus suggesting confounding by indication. Moreover, the association between ARB use and hypoglycaemia was negative in ADA users. PMID- 18507661 TI - Glucagon is absorbed from the rectum but does not hasten recovery from hypoglycaemia in patients with type 1 diabetes. AB - AIMS: A failure to secrete glucagon during hypoglycaemia is near universal in patients with type 1 diabetes 5 years after disease onset and may contribute to delayed counter-regulation during hypoglycaemia. Rectal glucagon delivery may assist glucose recovery following insulin-induced hypoglycaemia in such patients and has not been previously studied. METHODS: Six male patients (age 21-38 years) with type 1 diabetes (median duration 10 years) without microvascular complications, were studied supine after an overnight fast on two separate occasions at least 14 days apart. After omission of their usual morning insulin and 45 min rest, hypoglycaemia was induced by an intravenous insulin infusion which was terminated when capillary glucose concentration reached 2.5 mmol l(-1). Subjects were randomized to insert a rectal suppository containing 100 mg indomethacin alone (placebo) or 100 mg indomethacin plus 1 mg glucagon at the hypoglycaemic reaction. Serial measurements were made for 120 min. RESULTS: In the two groups, mean (SD) plasma glucose concentrations fell to a similar nadir of 1.8 (0.7) mmol l(-1) (placebo) and 2.1 (1.2) mmol l(-1) (glucagon). Peak plasma glucagon following hypoglycaemia was higher in the glucagon group; 176 (32) ng l(-1)vs. 99 (22) ng l(-1) after placebo (P = 0.006). However, the glucose recovery rate over 120 min after hypoglycaemia did not differ significantly. CONCLUSIONS: Our results provide evidence for the absorption of glucagon from the rectum. They also indicate that 1 mg does not constitute a useful mode of therapy to hasten recovery from hypoglycaemia in patients with type 1 diabetes. PMID- 18507662 TI - Haemoptysis associated with gatifloxacin in a 27 year old male asthmatic--a case report. PMID- 18507663 TI - Choice of first antihypertensive--comparison between the Irish and Dutch setting. PMID- 18507664 TI - Measuring learning from the TRC pharmacology E-Learning program. AB - AIM: Clinical pharmacology at the Leiden University Medical Centre is primarily taught by the Teaching Resource Centre's (TRC) Pharmacology database. The TRC program contains schematic graphics using a unique icon language, explanation texts and feedback questions to explain pharmacology as it pertains to pathophysiology. Nearly each course of the curriculum has a chapter in the TRC database offered for self-study. Since using the TRC program is not compulsory, the question remains whether students benefit from using it. METHODS: We compared the parameters of log-in attempts and time spent at each topic with students' final exam grades. Instead of looking at the regression of time spent on TRC on grade for one course, we looked at the individual student regression of time spent on TRC for different courses on grades. Spending more time using the TRC being associated with higher grades within an individual is a more powerful result than between students within a course, as better students are likely to spend more time using the TRC. RESULTS: Students increasingly used the program throughout the curriculum. More importantly, the time spent using the program showed that increased TRC use by an individual student is associated with a (small) increase in grade. As expected for a noncompulsory activity, better students (those with higher than average exam scores) logged in to the TRC more frequently, but poorer students appeared to have a larger benefit. CONCLUSIONS: An increase in TRC use by an individual student correlates with an increase in course grades. PMID- 18507665 TI - An antibody to the aggregated synthetic prion protein peptide (PrP106-126) selectively recognizes disease-associated prion protein (PrP) from human brain specimens. AB - Human prion diseases are characterized by the conversion of the normal host cellular prion protein (PrP(C)) into an abnormal misfolded form [disease associated prion protein (PrP(Sc))]. Antibodies that are capable of distinguishing between PrP(C) and PrP(Sc) may prove to be useful, not only for the diagnosis of these diseases, but also for a better understanding of the molecular mechanisms involved in disease pathogenesis. In an attempt to produce such antibodies, we immunized mice with an aggregated peptide spanning amino acid residues 106 to 126 of human PrP (PrP106-126). We were able to isolate and single cell clone a hybridoma cell line (P1:1) which secreted an IgM isotype antibody [monoclonal antibody (mAb P1:1)] that recognized the aggregated, but not the monomeric form of the immunogen. When used in immunoprecipitation assays, the antibody did not recognize normal PrP(C) from non-prion disease brain specimens, but did selectively immunoprecipitate full-length PrP(Sc) from cases of variant and sporadic Creutzfeldt-Jakob disease and Gerstmann-Straussler-Scheinker disease. These results suggest that P1:1 recognizes an epitope formed during the structural rearrangement or aggregation of the PrP that is common to the major PrP(Sc) types found in the most common forms of human prion disease. PMID- 18507666 TI - Lichen sclerosus et atrophicus responding to methotrexate. PMID- 18507667 TI - Immediate functional loading of implants in single tooth replacement: a prospective clinical multicenter study. AB - OBJECTIVES: The aim of the present study was to evaluate the outcome of immediate functional loading of implants in single-tooth replacement using two different installation procedures. MATERIAL AND METHODS: One hundred and fifty-one subjects, who required single-tooth rehabilitation in the area of 15-25 and 35 45, were enrolled in eight private clinics in Italy. The implant sites were randomly allocated to one of the following treatment groups. In the control group, in which a standard preparation procedure for implant placement and submerged healing of the implant was used, abutment connection and loading of the implants were performed 3 months after installation. In the test group 1, a standard preparation procedure for the implant placement and immediate functional loading of implant was carried out. In the test 2 group, however, a modified implant installation procedure (osteotome technique) was used followed by immediate functional loading of the implant. Clinical and radiographic examinations were performed at 3 and 12 months of follow-up at all sites. RESULTS: Three implants (5.5%) from the test 2 group (osteotome preparation) and one (2%) from the test 1 group (conventional drill preparation) failed to integrate and were removed one and three months after implant installation. The mean marginal bone loss assessed at 12 months was 0.31 mm (test 1), 0.25 mm (test 2) and 0.38 mm (control) (no statistically significant differences were found between the three treatment groups.) CONCLUSION: It is suggested that immediate functional loading of implants that are placed with a conventional installation technique and with sufficient primary stability may be considered as a valid treatment alternative in a single-tooth replacement. PMID- 18507668 TI - Capnovolumetry: a new tool for lung function testing in children with asthma. AB - In capnovolumetry, the expiratory CO2 concentration of exhaled air is plotted against the volume and thereby allows to determine functional dead space volumes. This method might offer additional information in lung function testing in children and adolescents with bronchial asthma. We aimed at determining whether a bronchospasmolysis (BSL) effect in the lower airways could also be detected by capnovolumetry as reflected by changes in the functional threshold dead space volumes (VDT). In 47 patients (aged 4-16 years) with a mild persistent bronchial asthma, VDT were determined before and after bronchodilation prior to starting therapy with inhaled steroids and after 6 months of treatment. Additionally, spirometry and body plethysmography were performed in all patients. There were significantly higher VDT values after BSL before and after 6 months of therapy (P<0.0001). VDT values before BSL were tendatively higher after 6 months of therapy compared with baseline values (P=0.07). VDT values correlated with parameters derived from conventional pulmonary function testing, i.e. vital capacity, forced expiratory volume in 1 s (FEV1), and maximum expiratory flow (MEF50). As VDT values particularly reflect the volumes of the lower bronchi this method may provide supplementary information to conventional lung function tests which are based on breathing mechanics. This seems to be especially helpful in situations where body plethysmography is not available or cooperation in forced expiration manoeuvres is insufficient. PMID- 18507669 TI - Recurrences in heart rate dynamics are changed in patients with diabetes mellitus. AB - Detection of subclinical autonomic dysfunction in patients with diabetes mellitus (DM) is of vital importance for risk stratification and subsequent management. Heart rate variability (HRV) analysis is a sensitive tool for assessment of cardiovascular autonomic dysfunction. As the heart is controlled by non-linear deterministic system, the non-linear dynamics measures should be preferred. Recurrence plot (RP) is able to analyse recurrences within system dynamics. The aim of the study was to detect heart rate dysregulation in DM by RP and to ascertain which of the recurrence quantification analysis (RQA) measures are changed in patients with DM compared to control group. We analysed HRV recordings from 17 young patients with type 1 DM and 17 healthy matched control subjects. RQA was performed on RPs with a fixed value of recurrence points percentage. From RQA measures based on diagonal lines, we have found higher percentage of determinism in DM group (P=0.038). Trapping time measure was also higher in DM (P=0.022). RQA revealed changes in dynamics recurrences with reduced complexity of heart rate control in young diabetic patients. As RQA parameters are independent of overall HRV, parameters of RP should be used together with linear HRV parameters for better description of heart rate dysregulation in patients with diabetics. PMID- 18507670 TI - Assessment of exercise capacity in women with type 2 diabetes. AB - The primary aim of this study was to compare the maximal oxygen uptake as evaluated from a submaximal exercise test (EVO2peak) to direct measurements of VO2peak during a maximal exercise test as means of monitoring the aerobic endurance capacity in women with type 2 diabetes (T2D). Twenty-seven women with T2D participated in the study. The program consisted of combined group training 1 h twice a week during 12 weeks and walks 1 h per week. EVO2 max was estimated using a submaximal exercise test on a bicycle ergometer ad modum Astrand. VO2peak and maximal work rate were measured using an incremental maximal exercise test on an electrically braked bicycle ergometer at baseline and after 6 and 12 weeks. EVO2peak was higher than VO2peak at baseline and significantly higher at 12 weeks (EVO2peak1.92+/-0.54 l min(-1), VO2peak 1.41+/-0.36, P<0.005). Maximal work rate increased significantly after 12 weeks (12+/-15, P<0.005) compared to baseline. The main finding of this study was that EVO2peak assessed using a submaximal exercise test, systematically overestimated VO2peak. The combined group training increased maximal work rate but not VO2peak. This is likely to reflect peripheral adaptation to exercise and/or improved mechanical efficiency. PMID- 18507671 TI - Escherichia coli O157 can grow in natural freshwater at low carbon concentrations. AB - Whereas much information on the die-off of Escherichia coli in the aquatic environment is available, only few data support its growth under such conditions. We therefore investigated batch growth in microcosms containing different types of sterile freshwater. The water samples were inoculated with low starting cell concentrations of E. coli O157 (3 x 10(3) cells ml(-1)) and growth was followed using nucleic acid staining combined with flow cytometry. We demonstrated that E. coli O157 is able to grow in sterile freshwater at low carbon concentrations, which is against the common view that cell numbers decline over time when added to freshwater samples. A correlation between apparent assimilable organic carbon (AOC(app)) concentration and the final cell concentration reached by E. coli O157 was established (P < 0.01). A considerable fraction of the AOC(app) (34 +/- 13%) was used by E. coli O157 but the numerical cell yield was about five-times lower in comparison with the bacterial AOC-test community, which originated from natural freshwater. On average, the maximum specific growth rate (mu(max)) of E. coli O157 growing in sterile freshwater at 30 degrees C was 0.19 +/- 0.07 h(-1). Batch growth assays at five different temperatures revealed a positive influence of temperature on mu(max) of E. coli O157. The results give new information on the behaviour of this common pathogen in the aquatic environment and contribute to microbial risk assessment in order to prevent spreading of water-borne diseases. PMID- 18507672 TI - The exopolysaccharide of Rhizobium sp. YAS34 is not necessary for biofilm formation on Arabidopsis thaliana and Brassica napus roots but contributes to root colonization. AB - Microbial exopolysaccharides (EPSs) play key roles in plant-microbe interactions, such as biofilm formation on plant roots and legume nodulation by rhizobia. Here, we focused on the function of an EPS produced by Rhizobium sp. YAS34 in the colonization and biofilm formation on non-legume plant roots (Arabidopsis thaliana and Brassica napus). Using random transposon mutagenesis, we isolated an EPS-deficient mutant of strain YAS34 impaired in a glycosyltransferase gene (gta). Wild type and mutant strains were tagged with a plasmid-born GFP and, for the first time, the EPS produced by the wild-type strain was seen in the rhizosphere using selective carbohydrate probing with a fluorescent lectin and confocal laser-scanning microscopy. We show for the fist time that Rhizobium forms biofilms on roots of non-legumes, independently of the EPS synthesis. When produced by strain YAS34 wild type, EPS is targeted at specific parts of the plant root system. Nutrient fluctuations, root exudates and bacterial growth phase can account for such a production pattern. The EPS synthesis in Rhizobium sp. YAS34 is not essential for biofilm formation on roots, but is critical to colonization of the basal part of the root system and increasing the stability of root-adhering soil. Thus, in Rhizobium sp. YAS34 and non-legume interactions, microbial EPS is implicated in root-soil interface, root colonization, but not in biofilm formation. PMID- 18507673 TI - Metabolic syndrome and three of its components as risk factors for recurrent ischaemic stroke presenting as large-vessel infarction. AB - BACKGROUND AND PURPOSE: Although a clear protocol for reduction of recurrent ischaemic stroke (RIS) has been established, few studies have compared the stroke subtype distribution and risk factors between RIS and first-ever stroke (FES). METHODS: This one-year hospital-based study enrolled 587 FES and 475 RIS patients. Patients were categorized into four stroke subtypes according to a modified TOAST stroke subtype classification system. Risk factor profiles were compared between the two major stroke groups and between the corresponding four subtypes to discriminate the significant risk factors for RIS. RESULTS: A multivariate regression analysis identified hypertension (OR, 1.87; 95% CI, 1.34 2.62), diabetes mellitus (DM) (OR, 1.57; 95% CI, 1.22-2.02), low high-density lipoprotein (LHDL) (OR, 1.43; 95% CI, 1.08-1.88) and older age as significant RIS risk factors. The significance of the former three RIS factors was further recognized in its large-vessel subtype. Moreover, metabolic syndrome was significantly more common in the recurrent stroke group (P = 0.01), including its large-vessel subtype (P = 0.04). Progressively increasing odds ratios from 1.49 to 2.02, in accordance with increased number of diagnostic components of metabolic syndrome for recurrent large-vessel ischaemic stroke, were noted. CONCLUSIONS: Metabolic syndrome likely plays a crucial role in the development of RIS, including large-vessel infarction in modern-day Taiwan. PMID- 18507674 TI - Aortic calcification on plain chest radiography predicts embolic complications during carotid artery stenting. AB - BACKGROUND AND PURPOSE: Aortic arch calcification (AC) on conventional angiograms has recently been shown to be associated with embolic complications during carotid artery stenting (CAS). Because conventional angiography is not routinely carried out in patients prior to CAS, a less invasive, commonly available diagnostic modality is warranted to identify AC. METHODS: We investigated the occurrence of AC on routinely acquired chest radiographs and its influence on diffusion-weighted imaging (DWI) lesions as surrogate markers for stroke. RESULTS: A total of 152 patients (mean age: 67.5 +/- 8.9 years, 112 men) underwent CAS and completed pre- and post-procedural DWI. AC larger than 1 cm in length was classified as relevant and could be detected in 63 patients (41.4%) on plain chest radiographs. In patients with AC, significantly more new DWI-lesions were found than in patients without AC [median 2; interquartile range (IQR): 0-7 vs. median 1, IQR: 0-2; P < 0.05]. After multivariate regression analysis, AC was independently associated with new post-procedural DWI lesions. DISCUSSION: AC is a common finding in patients with a high-grade carotid stenosis and predicts embolic complications during CAS. Since AC can be detected on plain chest radiography, this diagnostic tool is useful to identify high-risk patients for CAS. PMID- 18507675 TI - Voice tremor in monozygotic twins. PMID- 18507676 TI - Intraoperative microrecording under general anaesthesia with bispectral analysis monitoring in a case of deep brain stimulation surgery for Parkinson's disease. PMID- 18507677 TI - 'Subclinical MS': follow-up of four cases. AB - BACKGROUND AND PURPOSE: Subclinical multiple sclerosis (S-MS) can be defined as the discovery of characteristic lesions at magnetic resonance (MR) or at autopsy, in the absence of clinical evidence consistent with MS. To describe four cases of S-MS cases followed-up with clinical and MR examinations. METHODS: The four patients were initially evaluated because of accidental MRI findings suggestive of MS that fulfilled the Barkhof criteria. At the moment of MR examination, patients were asymptomatic or complained of unspecific symptoms. The objective examinations as well as the clinical history were negative. An extensive diagnostic work-up excluded alternative diagnoses. RESULTS: Case 1: 40-year woman undergoing MR examination due to trivial cervical trauma. The cerebrospinal fluid (CSF) analysis showed an IgG Index of 3.32 and presence of oligoclonal bands (OCB). New MR enhancing lesions were detected in the brain and cervical cord after 2 and 5 years. The patient remained clinically asymptomatic. Neuropsychological assessment (NPS) performed after 5 years revealed moderate deficits of attention/concentration and executive functions. Case 2: 35- year man, undergoing MR due to recent-onset headache. CSF analysis showed an IgG Index of 1.54 and presence of OCB. At baseline, NPS assessment revealed moderate deficit on complex attention tasks. New MR enhancing lesions were detected in the brain after 1 and 3 years. Four years after the first MR, the patient presented double vision with internuclear ophthalmoplegia. He fulfilled Polman's criteria for MS and he started interferon beta therapy. He remained stable at a 8-year follow-up. Case 3: 32-year man, undergoing MR due to brief loss of consciousness following neck hyperextension. CSF analysis showed absence of OCB and normal IgG Index. At baseline, NPS assessment revealed mild deficits of attention and executive functions. Over a 5-year follow-up the subject remained asymptomatic and MR was unchanged. Case 4: 40-year woman, sister of an MS patient. MR was required during a period of marked tiredness taking into account the family history. The patient did not present any relevant symptoms and MRI remained unchanged over a 6-year follow-up. NPS evaluation performed after 7 years showed moderate impairment of memory, attention and executive functions. CONCLUSION: With the increased use of MR, S-MS cases can be identified more often. In our series, despite the absence of any typical symptoms, an NPS assessment detected cognitive impairments consistent with those reported in MS. PMID- 18507678 TI - Assignment of Staphylococcus aureus isolates to clonal complexes based on microarray analysis and pattern recognition. AB - A DNA microarray was designed for the rapid genotyping of Staphylococcus aureus. It covers 185 distinct genes and about 300 alleles thereof, including species specific controls, accessory gene regulator (agr) alleles, genes encoding virulence factors, and microbial surface components recognizing adhesive matrix molecules, capsule type-specific genes, as well as resistance determinants. It was used to examine 100 clinical isolates and reference strains. Relationships of leukocidin and ssl/set (staphylococcal superantigen-like or exotoxin-like) genes were reviewed considering these experimental results as well as published sequences. A good correlation of overall hybridization pattern and multilocus sequence typing was found. Analysis of hybridization profiles thus allowed not only to assess virulence and drug resistance, but also to assign isolates to strains and to clonal complexes. Hybridization data were used to construct a split network tree and to analyse relationships between strains. Allelic variations of a number of genes indicate a division of S. aureus into three major branches that are not in accordance to agr group or capsule-type affiliations. Additionally, there are some isolated lineages, such as ST75, ST93, or ST152. These strains produce aberrant hybridization profiles, indicating that only a part of the gene pool of S. aureus has been investigated yet. PMID- 18507680 TI - Progress towards understanding the fate of plasmids in bacterial communities. AB - Plasmid-mediated horizontal gene transfer influences bacterial community structure and evolution. However, an understanding of the forces which dictate the fate of plasmids in bacterial populations remains elusive. This is in part due to the enormous diversity of plasmids, in terms of size, structure, transmission, evolutionary history and accessory phenotypes, coupled with the lack of a standard theoretical framework within which to investigate them. This review discusses how ecological factors, such as spatial structure and temporal fluctuations, shape both the population dynamics and the physical features of plasmids. Novel data indicate that larger plasmids are more likely to be harboured by hosts in complex environments. Plasmid size may therefore be determined by environmentally mediated fitness trade-offs. As the correlation between replicon size and complexity of environment is similar for plasmids and chromosomes, plasmids could be used as tractable tools to investigate the influence of ecological factors on chromosomes. Parallels are drawn between plasmids and bacterial facultative symbionts, including the evolution of some members of both groups to a more obligate relationship with their host. The similarity between the influences of ecological factors on plasmids and bacterial symbionts suggests that it may be appropriate to study plasmids within a classical ecological framework. PMID- 18507679 TI - Enhanced resistance against systemic Candida albicans infection in mice treated with C. albicans DNA. AB - In this study, double-stranded Candida albicans DNA was administered in systemic C. albicans infection in at dose of 20 microg per mouse at 4, 5 and 6 weeks of age. The level of IL-12 in serum was elevated as a result of yeast DNA treatment and correlated with lower mortality and decreased kidney and liver injury. Macrophage activation was demonstrated by an increase of nitric oxide (NO) and IL 12 production. These effects were Janus activation kinases (JAK)/signal transducer and activator of transcription (STAT) dependent as they were inhibited by selective JAK inhibitor tyrphostin AG-490. DNA influenced adaptive immune response through elevation of anti-Candida IgG antibody production in systemic C. albicans infection. Thus, C. albicans DNA augmented innate and adaptive immune responses against the pathogen. PMID- 18507681 TI - Microdiversity of Burkholderiales associated with mycorrhizal and nonmycorrhizal roots of Medicago truncatula. AB - The genetic diversity of bacterial communities associated with mycorrhizal and nonmycorrhizal roots of Medicago truncatula was characterized by two approaches. Firstly, phylogenetic analysis was performed on 164 partial 16S rRNA gene intergenic spacer (IGS) sequences from operational taxonomic units previously shown to be preferentially associated with mycorrhizal roots. These sequences were distributed into three branches corresponding to Comamonadaceae, Oxalobacteraceae and Rubrivivax subgroups. Most sequences were obtained from mycorrhizal roots, indicating the preferential association of the corresponding families with mycorrhizal roots. A second phylogenetic analysis was performed on the partial 16S rRNA gene-IGS sequences of 173 isolates among a large collection of isolates, from mycorrhizal and nonmycorrhizal roots, belonging to Comamonadaceae and Oxalobacteraceae on the basis of their positive hybridization with a partial 16S rRNA gene-IGS probe obtained in this study. Sequence analysis confirmed the affiliation of 166 isolates to Comamonadaceae and seven to Oxalobacteraceae. Oxalobacteraceae isolates were more abundant in mycorrhizal (five) than in nonmycorrhizal (two) roots, whereas Comamonadaceae isolates were more abundant in nonmycorrhizal (109) than mycorrhizal roots (57). Further analysis of Comamonadaceae isolates by BOX-PCR showed that the genetic structure of culturable populations belonging to this family differed significantly in mycorrhizal and nonmycorrhizal roots, as indicated by distributions in different BOX types, differences being significantly explained by BOX types only including isolates from mycorrhizal roots. These data are discussed in an ecological context. PMID- 18507682 TI - Evaluation of a multiplex PCR for detection of serotypes K1, K2 and K5 in Klebsiella sp. and comparison of isolates within these serotypes. AB - A multiplex PCR using targets within the serotype-specific region of the capsular polysaccharide synthesis gene cluster of serotypes K1, K2 and K5 was evaluated using the 77 reference serotype strains of Klebsiella, and a panel of clinical isolates subjected previously to conventional serotyping. The PCR was highly specific for these serotypes, which are those most associated with virulence in humans and horses. PCR confirmed that isolates of the K5 serotype had cross reacted with antiserum for other serotypes, particularly for K7. K5 isolates received by our laboratory were almost exclusively from thoroughbred horses, and were submitted for screening prior to breeding programmes. Most, including a reference strain isolated in 1955, belonged to a cluster of genetically similar isolates of sequence type (ST) 60. K1 isolates, all from humans, belonged to a previously identified cluster of ST 23. PMID- 18507683 TI - The Escherichia coli biofilm-promoting protein Antigen 43 does not contribute to intestinal colonization. AB - Abstract Escherichia coli is a versatile organism capable of causing a variety of intestinal and extraintestinal diseases, as well as existing as part of the commensal flora. A variety of factors permit specific attachment to host receptors including fimbrial adhesins and outer membrane proteins such as autotransporters. One of the better characterized autotransporters is Antigen 43 (Ag43), the major phase-variable surface protein of E. coli. Ag43 is associated with bacterial cell-cell aggregation and biofilm formation. Nevertheless, the precise biological significance and contribution to intestinal colonization remain to be elucidated. Here we investigated the contribution of Ag43 to E. coli adherence to intestinal epithelial cells and colonization of the mouse intestine. These investigations revealed that Ag43 increased in vitro adherence of E. coli to epithelial cells by promoting bacterial cell-cell aggregation but that Ag43 did not promote specific interactions with the mammalian cells. Furthermore, Ag43 did not contribute significantly to colonization of the mouse intestine and expression of Ag43 was lost a few days after colonization of the mouse was established. Unexpectedly, considering its similarity to other adhesins, our findings suggest that Ag43 does not act as a direct colonization factor by binding to mammalian cells. PMID- 18507684 TI - The rpoE operon regulates heat stress response in Burkholderia pseudomallei. AB - Burkholderia pseudomallei is a gram-negative bacterium and the causative agent of melioidosis, one of the important lethal diseases in tropical regions. In this article, we demonstrate the crucial role of the B. pseudomallei rpoE locus in the response to heat stress. The rpoE operon knockout mutant exhibited growth retardation and reduced survival when exposed to a high temperature. Expression analysis using rpoH promoter-lacZ fusion revealed that heat stress induction of rpoH, which encodes heat shock sigma factor (sigma(H)), was abolished in the B. pseudomallei rpoE mutant. Analysis of the rpoH promoter region revealed sequences sharing high homology to the consensus sequence of sigma(E)-dependent promoters. Moreover, the putative heat-induced sigma(H)-regulated heat shock proteins (i.e. GroEL and HtpG) were also absent in the rpoE operon mutant. Altogether, our data suggest that the rpoE operon regulates B. pseudomallei heat stress response through the function of rpoH. PMID- 18507685 TI - Effect of primary mild stresses on resilience and resistance of the nitrate reducer community to a subsequent severe stress. AB - The factors regulating soil microbial stability (e.g. resistance and resilience) are poorly understood, even though microorganisms are essential for ecosystem functioning. In this study, we tested whether a functional microbial community subjected to different primary mild stresses was equally resistant or resilient to a subsequent severe stress. The nitrate reducers were selected as model community and analysed in terms of nitrate reduction rates and genetic structure by narG PCR-restriction fragment length polymorphism fingerprinting. Heat, copper and atrazine were used as primary stresses and mercury at a high concentration as a severe stress. None of the primary stresses had any significant impact on the nitrate reducer community. Although primary stress with heat, copper or atrazine had no effect on the resilience of the nitrate reducer activity to mercury stress, pre-exposure to copper, another heavy metal, resulted in increased resilience. In contrast, the resistance of both structure and activity of the nitrate reducer community to severe mercury stress was not affected by any of the primary stresses tested. Our experiment suggests that the hypothetical effect of an initial stress on the response of a microbial community to an additional stress is complex and may depend on the relatedness of the two consecutive stresses and the development of positive cotolerance. PMID- 18507686 TI - Tumor necrosis factor-alpha induces the aberrant expression of mucus core protein 2 in non-neoplastic biliary epithelial cells via the upregulation of CDX2 in chronic cholangitis. AB - AIM: Chronic cholangitis, such as hepatolithiasis, is frequently associated with goblet cell metaplasia and the aberrant expression of mucus core protein-2 (MUC2). In this study, we clarified the role of inflammatory cytokines in the expression of MUC2 in lining biliary epithelial cells (BEC) in chronic cholangitis with an emphasis on CDX2, an intestine-specific transcription factor. METHODS: We used human hepatolithiatic livers and polycystic kidney (PCK) rats, an animal model of Caroli's disease, and cultured BEC from PCK rats. As a control, extrahepatic biliary obstruction and histologically normal livers and Crj:CD rats were used. RESULTS: Immunohistochemically, tumor necrosis factor alpha (TNF-alpha) was expressed in periductal inflammatory cells and BEC of the affected intrahepatic bile ducts with an aberrant expression of MUC2 and CDX2 in hepatolithiasis and the PCK rats. In cultured BEC, TNF-alpha, interleukin (IL) 1beta, IL-6, and interferon-gamma induced the expression of CDX2 mRNA, though only TNF-alpha additionally induced the expression of MUC2 mRNA. The expression of CDX2 mRNA and the MUC2 protein induced in BEC by TNF-alpha were abolished by pretreatment of nuclear factor-kappaB inhibitors. CONCLUSION: The aberrant expression of CDX2 and MUC2 in the affected bile ducts showing goblet cell metaplasia was closely associated with TNF-alpha expressed in periductal infiltrating inflammatory cells and BEC. TNF-alpha induced the expression of CDX2 and MUC2 in cultured BEC. Taken together, it seems likely that TNF-alpha plays a role in MUC2 expression via CDX2 upregulation in the bile ducts with chronic cholangitis and goblet cell metaplasia. PMID- 18507687 TI - Primary hepatic presentation of Hodgkin's lymphoma: A case report. AB - Hodgkin's lymphoma (HL) is in general a lymph node-based disease. Hepatic involvement usually occurs in the advanced disease. Primary and prominent manifestation of the disease in the liver is extremely rare. We report magnetic resonance imaging leading to diagnosis in a rare case of liver involvement as the first sign of HL. PMID- 18507688 TI - Stones from cancerous and benign gallbladders are different: A proton nuclear magnetic resonance spectroscopy study. AB - AIM: Gallbladder cancer (GBC) is frequently associated with gallstones (GS). At the same time, however, a very small number of patients with GS develop GBC. Cholesterol and metal salts are the common constituents of all GS. To understand their role in the etiopathogenesis of GBC, cholesterol, calcium, and magnesium composition in GS is compared in cancerous and benign gallbladders. METHODS: GS from patients with GBC (n = 11), chronic cholecystitis (CC; n = 23), and xanthogranulomatous cholecystitis (XGC; n = 11) undergoing cholecystectomy were analyzed using proton nuclear magnetic resonance spectroscopy. The diagnosis of the gallbladder disease was based on histopathological examinations. Cholesterol, calcium, and magnesium in the GS of GBC, XGC, and CC were analyzed, compared, and correlated using statistical methods. RESULTS: The quantity of cholesterol was significantly less in the GS of GBC than in benign gallbladder diseases (CC or XGC, P < 0.0001 for both). Both calcium and magnesium were significantly higher in GBC than in benign disease (calcium: P < 0.0005 and magnesium: P < 0.0001 for GBC vs CC; calcium: P < 0.02 and magnesium: P < 0.04 for GBC vs XGC). In all the GS, calcium was higher than magnesium. Calcium and magnesium were positively correlated in GBC (R = 0.69) and XGC (R = 0.75), and cholesterol and calcium were negatively correlated in CC (R =-0.61). CONCLUSION: Differences in the GS composition between malignant and benign gallbladder patients may provide useful clues to the etiopathogenesis of GBC. These clues could lead to the identification of patients with GS in vivo who are at high risk of developing GBC, and advocate prophylactic cholecystectomy to prevent GBC. PMID- 18507689 TI - Does a late evening meal reduce the risk of hepatocellular carcinoma among patients with chronic hepatitis C? AB - AIM: Some studies have suggested that nutritional support might protect against the recurrence of hepatocellular carcinoma (HCC) among postoperative HCC patients. However, no epidemiological studies have evaluated the effect of nutritional support on HCC incidence. This study aimed to investigate the association between a late evening meal and HCC. METHODS: We conducted a hospital based, case-control study comparing 73 cases with HCC to 253 matched controls among patients with chronic hepatitis C. A questionnaire elicited information on the consumption of a late evening meal, which was defined as a snack or meal within 2 h before bedtime. The odds ratios (OR) and 95% confidence intervals (CI) were calculated by the conditional logistic regression model. RESULTS: After adjustment for potential confounders, patients who consumed a late evening meal had a lower OR as compared to those who did not consume one (OR, 0.08; 95% CI, 0.01-0.48). In terms of frequency of intake, a clear inverse exposure-response relationship was observed (trend P = 0.009). In addition, a negative association between a late evening meal and HCC was more pronounced among patients with an alpha-fetoprotein level of less than 20 ng/mL and those with a body mass index of less than 25 kg/m(2). CONCLUSION: A late evening meal might protect against HCC, particularly among patients with a normal alpha-fetoprotein level and who are not obese, although these relations might be accounted for other factors, including total energy intake. Further studies with larger study sizes are needed to corroborate these findings. PMID- 18507690 TI - Current studies on therapeutic approaches for ischemia/reperfusion injury in steatotic livers. AB - Steatotic livers are particularly vulnerable to ischemia/reperfusion (I/R) injury, resulting in poor outcomes following liver surgery and transplantation. Therapeutic approaches for I/R injury in steatotic livers are currently under intensive investigation. This review summarizes and discusses the approaches developed during the last few years to prevent hepatic I/R injury in steatotic livers. Among the proposed approaches, ischemic preconditioning and intermittent clamping are the two most promising approaches that have been applied in some clinical centers for liver surgery and transplantation, but most of others have not reached clinical application yet. PMID- 18507691 TI - Abstinence in patients with alcoholic liver cirrhosis: A follow-up study. AB - AIM: To investigate the proportion of patients with alcoholic cirrhosis who abstained from alcohol after contact with a hepatology unit, the predictors for abstinence, and the role of clinical and psychosocial factors in short-term mortality in these patients. METHODS: Eighty-seven consecutive patients with alcoholic cirrhosis from a transplant center were included. Data on cirrhosis severity and complications, as well as on abstinence and psychosocial factors were collected. Patients were followed up for 19 (12-25) months. Data on abstinence during follow up, alcohol abuse treatment, psychiatric contact, severity of cirrhosis, mortality, and liver transplantation were analyzed. RESULTS: Prior to inclusion, 53/87 (61%) patients had abstained from alcohol for 24 months (interquartile range: 18-33). Twenty percent had a history of other substance abuse, 47% had undergone alcohol abuse treatment, and 21% had a previous psychiatric diagnosis. Forty-eight percent lived with a partner, 23% worked/studied, and 53% were pensioners. During follow up, 26% died, 20% received a liver transplant, 55% abstained from alcohol, 47% received alcohol abuse treatment, and 33% had psychiatric contact. In a multivariate analysis, abstinence during follow up was found to be related to abstinence upon inclusion in the study, to the model for end-stage liver disease (MELD) score at follow up, and to no abuse treatment in a detoxification unit, whereas mortality was related to index MELD and alcohol abuse treatment during follow up. Neither abstinence nor mortality was related to psychosocial factors. CONCLUSION: More than half of patients with alcoholic cirrhosis were found to abstain from alcohol during follow up, which was related to prior documentation of abstinence and cirrhosis severity. Cirrhosis severity (expressed as the MELD) and alcohol abuse treatment during follow up were related to short-term mortality. PMID- 18507692 TI - Potential of laparoscopy in chronic liver disease with hepatitis B and C viruses. AB - AIM: The definitive diagnosis of chronic liver disease is made either by a histological examination of a biopsy specimen or upon visualization of the liver surface at laparoscopy. The aim of this retrospective cohort study is to assess whether histological or laparoscopic findings are associated with hepatocellular carcinoma (HCC) development. METHODS: A retrospective review of paired laparoscopy and histology reports was performed on 4124 hepatitis virus-positive patients who underwent laparoscopy: 2804 patients had hepatitis C virus (HCV group) and 1320 patients had hepatitis B virus (HBV group). Based on the irregularities of the liver surface, the laparoscopic findings were classified into three groups in progression order: smooth, irregular, or nodular. The histological findings were classified according to the extent of fibrosis into four stages (stages 1-4) in progression order. RESULTS: The number of patients with HCC development was 565 in the HCV group and 115 in the HBV group. The Coxregression hazard model showed that HCC appearance in the HCV group was independently associated with laparoscopic findings (relative risk based on every progression of one rank [RR], RR = 4.31, P < 0.0001) and histological findings (RR = 2.56, P < 0.0001). In the HBV group, however, HCC appearance in was mainly associated with laparoscopic findings (RR = 2.12, P < 0.0001) compared to histological findings (RR = 1.13, P = 0.403). CONCLUSION: Our data indicate that laparoscopic findings of the liver are dominant predictors for HCC development compared with histological findings in patients with HBV. PMID- 18507693 TI - Blockade of Rho/Rho-associated coiled coil-forming kinase signaling can prevent progression of hepatocellular carcinoma in matrix metalloproteinase-dependent manner. AB - AIM: There is growing evidence that the Rho/Rho-associated coiled coil-forming kinase (ROCK) signaling pathway is upregulated in tumors and plays a key role in cancer invasion and metastasis. Our aim was to test the anticancer effects of Rho/ROCK inhibitor, Y-27632, including possible mechanisms in a highly metastasizing hepatocellular carcinoma (HCC) mouse model on its secretion of matrix metalloproteinase (MMP) and tumor progression. METHODS: Following orthotopic implantation of CBO140C12 HCC tumor fragments into the liver of mice, the mice were randomly assigned to a Y-27632-treated group or control group. After treatment for 4 weeks, specimens were obtained to evaluate tumor size, metastases, and immunohistochemical findings. In vitro, we examined the effects of Y-27632 and RhoC siRNA on MMP-2 and -9 expressions, invasiveness, and apoptosis in cultured tumor cells. RESULTS: Both RhoA and RhoC were upregulated in HCC-bearing livers, and Y-27632 significantly inhibited not only tumor growth and intrahepatic metastasis (P < 0.05), but also tumoral MMP-9 expression. Moreover, Y-27632 treatment resulted in large necrotic areas in tumors. In vitro, Y-27632 and RhoC siRNA reduced MMP-2 and -9 expressions, as well as the chemotactic migration of tumor cells dose-dependently, and increased apoptosis eight times. CONCLUSION: Y-27632 suppresses progression and limits the intrahepatic metastasis of established HCC. This could be linked to the decreased MMP expression and induction of apoptosis in tumor cells. Rho signaling may prove to be a productive target in anticancer therapy. PMID- 18507694 TI - Analysis of morphological vascular changes of hepatocellular carcinoma by microflow imaging using contrast-enhanced sonography. AB - AIM: To determine whether the findings of microflow imaging (MFI), composed of a flash replenishment and a maximum intensity holding sequence, using contrast enhanced sonography, correlate with the degree of histological differentiation of hepatocellular carcinoma (HCC). METHODS: This study was approved by the institutional review board; patients gave informed consent. The samples comprised of 61 nodules histologically diagnosed as HCC: 20 well-differentiated, 26 moderately-differentiated, and 15 poorly-differentiated HCC. SonoVue was used as the ultrasound (US) contrast agent. The US equipment used was a SSA-770 A with the imaging mode set at MFI. MFI is an imaging method combining flash replenishment imaging and maximum intensity holding. Two independent readers (readers 1 and 2) classified the microflow images into four patterns: (i) normal pattern; (ii) cotton pattern; (iii) vascular pattern; and (iv) dead wood pattern. The results were compared with the degree of histopathological differentiation of the HCC. RESULTS: In each of the 61 HCC, blood vessels in the tumor were clearly resolved down to their fine branches. With regard to the relationship between imaging patterns and thehistological findings, it was found (with high percentages) that the normal and cotton patterns were associated with well differentiated HCC, that the vascular pattern was associated with moderately differentiated HCC, and that the dead wood pattern was associated with poorly differentiated HCC. If HCC with the normal and cotton patterns were assessed as well differentiated and those with the vascular or dead wood pattern were assessed as moderately or poorly differentiated, the sensitivity, specificity, and accuracy of these assessments were found to be 85%, 92.7%, and 90%, respectively, for reader 1, and 85%, 82.9%, and 83.6%, respectively, for reader 2. CONCLUSION: The angioarchitecture and hemodynamics of HCC could be evaluated in detail using MFI. The results of this study demonstrate the feasibility of a non-invasive preoperative diagnosis of the histological differentiation of HCC using MFI. PMID- 18507695 TI - The relative importance of resources and natural enemies in determining herbivore abundance: thistles, tephritids and parasitoids. AB - 1. The relative importance of host-plant resources and natural enemies in influencing the abundance of insect herbivores was investigated in potted plant and natural population experiments, using tephritid (Diptera: Tephritidae) flies, their host plant, creeping thistle Cirsium arvense, and their Hymenoptera parasitoids. 2. Experimental manipulation of host-plant quality (i.e. levels of host-plant nutrients) and resource availability (i.e. the number of buds) increased tephritid abundance. There was no evidence that the seed-feeding tephritid fly Xyphosia miliaria preferentially oviposited on fertilized C. arvense. 3. At low thistle densities, X. miliaria showed a constant rate of resource exploitation. At higher thistle densities, a threshold was detected, above which additional buds were not attacked. 4. Parasitism attack was variable across host (tephritid) densities but levels of parasitism were consistently higher on the fertilized thistles. 5. Experimental manipulation of host-plant quality and resource availability (quantity) not only directly affects the tephritid population but also, indirectly, leads to high rates of parasitism. Both chemical and physical characteristics of host plants affect the performance of natural enemies. 6. Both top-down and bottom-up forces act to influence tephritid abundance, with bottom-up influences appearing to be the most important. PMID- 18507696 TI - Long-term variation of link strength in a simple benthic food web. AB - 1. The predatory isopod Saduria entomon (L.) and its amphipod prey Monoporeia affinis (Lindstrom) are key components of the food web in the northern Baltic Sea, together representing 80-90% of the macrobenthic biomass. We use 20 years of stomach content data for Saduria to investigate how diet dynamics affect the stability of the interaction between Saduria and Monoporeia. 2. Consumption of the main prey, Monoporeia, fitted a type III functional response. Consumption rates of the most important alternative prey, mysids, were found to be unrelated to mysid densities but negatively related to the density of Monoporeia. The fit of consumption data to a model that assumes passive prey selection was poor. Thus we conclude that some form of active choice is involved. 3. The effect of consumption of mysids, the alternative prey, on the stability of this system was investigated using a 'one predator-two prey' model with stochastic environmental variation. Analysis of the model suggests that feeding on mysids leads to a decreased extinction risk for the predator, Saduria, and reduced density oscillations for both Saduria and its main prey, Monoporeia. PMID- 18507697 TI - Plasma progesterone, oestradiol-17beta and total oestrogen profiles in relation to oestrous behaviour during induced ovulation in Murrah buffalo heifers. AB - The objectives of this study were to establish the characteristics of oestrous behaviour in Ovsynch (induction of ovulation through administration of GnRH-PGF2 GnRH in a systemic manner on 0, seventh and ninth day respectively) and Ovsynch plus Norprolac (Quinagolide hydrochloride - an inhibitor of prolactin secretion) treated Murrah buffalo heifers and to determine the relationships between this behaviour and the plasma concentrations of oestradiol-17beta (E2), total oestrogen, and progesterone. Oestrus was detected by visual observations of oestrus signs, per rectal examination of genitalia and bull parading thrice a day during treatment period. Among all the symptoms, it was observed that bull mounting of heifers in oestrus was highest. Examination of genital tracts per rectum revealed that the cervix was relaxed, uterus was turgid and ovaries had palpable follicle in animals with oestrus. The peak concentrations of E2 (10.81 +/- 0.62 pg/ml) and total oestrogen (17.11 +/- 1.21 pg/ml) occurred at 9.45 +/- 0.85 and 9.64 +/- 0.93 h after second GnRH administration, respectively, in Ovsynch treated animals. However, the peak levels of E2 (20.02 +/- 2.87 pg/ml) and total oestrogen (32.71 +/- 3.15 pg/ml) occurred at 10.18 +/- 0.50 and 10.36 +/- 0.75 h after second GnRH administration, respectively, in Ovsynch plus Norprolac treated animals. Plasma progesterone concentration was basal (0.20 +/- 0.001 ng/ml) during the peri-oestrus period. The plasma progesterone concentration was the lowest on the day of oestrus and increased to register a peak on day 13 +/- 2 of the cycle. Oestrous behaviour was positively correlated with the peak concentration of E2 (p < 0.001) and total oestrogen (p < 0.001) during the peri-oestrus period. Inhibition of prolactin by Norprolac administration significantly increased the concentration of E2 and total oestrogen during oestrus in buffaloes in comparison to those recorded in animals subjected to Ovsynch protocol alone. In conclusion, our results suggest that the peak concentrations of E2 and total oestrogen and mean level of E2 and total oestrogen during the peri-oestrus period are the important factors contributing the behavioural manifestation of oestrus in buffalo cows. PMID- 18507698 TI - Loss of non-steroidal anti-inflammatory drug hypersensitivity after Helicobacter pylori eradication. PMID- 18507700 TI - The contribution of parasitism to selection on floral traits in Heuchera grossulariifolia. AB - Parasites are ubiquitous and have well-documented ecological consequences. In contrast, the extent to which parasites drive phenotypic evolution in hosts remains obscure. We use a recently developed statistical technique--selective source analysis--to analyse the strength of phenotypic selection acting on floral traits in the plant Heuchera grossulariifolia attributable to attack by the seed parasitic moth, Greya politella. This analysis spanned 3 years and included two sympatric populations of the host plant H. grossulariifolia that differ in ploidy. Our analyses revealed that attack by G. politella contributed to phenotypic selection for flowering time and floral display size, favouring earlier flowering in the polyploid population, later flowering in the diploid population and increased floral display size in the polyploid population. Although selection imposed by parasite attack was generally quite weak, in one of the 3 years parasites generated a modestly strong selection gradient (beta = 0.059) that explained 38.6% of total observed phenotypic selection for earlier flowering in the polyploid population. Together, our results demonstrate parasites can generate significant phenotypic selection, but that such selection may be sporadic across populations and time. PMID- 18507701 TI - Maintenance of clinal variation for shell colour phenotype in the flat periwinkle Littorina obtusata. AB - Clines can signal spatially varying selection and therefore have long been used to investigate the role of environmental heterogeneity in maintaining genetic variation. However, clinal patterns alone are not sufficient to reject neutrality or to establish the mechanism of selection. Indirect, inferential methods can be used to address neutrality and mechanism, but fully understanding the adaptive significance of clinal variation ultimately requires a direct approach. Ecological model systems such as the rocky intertidal provide a useful context for direct experimentation and can serve as a complement to studies in more traditional genetic model systems. In this study, we use indirect and direct approaches to investigate the role of environmental heterogeneity in the maintenance of shell colour polymorphism in the flat periwinkle snail, Littorina obtusata. We document replicated clines in shell colour morph frequencies over thermal gradients at two spatial scales, contrasting with patterns at previously reported microsatellite loci. In addition, experimental results demonstrate that that shell colour has predictable effects on shell temperature and that these differences in temperature, in turn, coincide with patterns of survivorship under episodic thermal stress. Direct manipulation of shell colour revealed that shell colour, and not a correlated character, was the target of selection. Our study provides evidence that spatially varying selection via thermal regime contributes to the maintenance of shell colour phenotype variation in L. obtusata in the sampled areas of the Gulf of Maine. PMID- 18507702 TI - Socioeconomic and psychological variables as risk and protective factors for parental well-being in families of children with intellectual disabilities. AB - BACKGROUND: The first aim of the present study was to estimate the extent to which differences in well-being in parents of children with and without intellectual disability (ID) in Sweden can be accounted for by differences in the presence of the risk factors: (1) child disability; (2) socioeconomic disadvantage; (3) household composition; and (4) parental characteristics. The second aim was concerned with individual variation in well-being within the group of parents of children with ID. The aim was to estimate if protective factors such as parental personality characteristics (sense of coherence), perceived positive impact of the child and satisfaction with participation in different arenas of life explained variation in well-being in mothers and fathers of children with ID over and above that explained by the risk factors. METHOD: Parents of children with ID (62 mothers and 49 fathers) and control children (183 mothers and 141 fathers) completed postal surveys on well-being, socioeconomic situation, health, sense of coherence, satisfaction with participation in different arenas of life and the child's impact on the family. RESULTS: The results showed that mothers of children with ID had lower levels of well-being than fathers and control parents, but the presence of a child with ID did not in itself predict poorer maternal well-being. Rather, differences in economic hardship and self-rated health were the strongest predictors for well-being. It was further found that 67.7% of the mothers of children with ID scored within the high well-being group. The predictive power of the model increased significantly for both fathers and mothers when protective factors were added to the model (42 and 78% explained variance compared with 25% with only risk factors). CONCLUSIONS: Well-being of parents with a child with ID is dependent upon the interplay of risk and protective factors and research needs to address these variables simultaneously. PMID- 18507703 TI - The relationship between sources and functions of social support and dimensions of child- and parent-related stress. AB - BACKGROUND: In this longitudinal study, we examined the relationship between the sources and functions of social support and dimensions of child- and parent related stress for mothers of young children with mild developmental delays. METHODS: Sixty-three mothers completed assessments of stress and support at two time points. RESULTS: Multiple regression analyses revealed that parenting support during the early childhood period (i.e. advice on problems specific to their child and assistance with child care responsibilities), irrespective of source, consistently predicted most dimensions of parent stress assessed during the early elementary years and contributed unique variance. General support (i.e. primarily emotional support and validation) from various sources had other, less widespread effects on parental stress. CONCLUSIONS: The multidimensional perspective of the construct of social support that emerged suggested mechanisms mediating the relationship between support and stress and provided a framework for intervention. PMID- 18507704 TI - Enzootic simian piroplasm (Entopolypoides macaci ) in wild-caught Kenyan non human primates. AB - BACKGROUND: Three species of non-human primates comprising African green monkeys (AGMs), (Cercopithecus aethiops, n = 89), Syke's monkeys (Cercopithecus mitis, n = 60) and olive baboons (Papio cynocephalus anubis, n = 30), were screened for Entopolypoides macaci. METHODS: Observation of blood smears prepared from these animals revealed E. macaci infection rate of 42.7% in AGMs, 35% in Syke's monkeys and 33.3% in baboons. RESULTS: Gender infection rate was 38.2% in females and 29% in males. Statistically, there was no significant difference in infection rates between the monkey species and sexes (P > 0.05). Subsequent indirect immuno fluorescent antibody test supported the morphological appearance of E. macaci observed by microscopy. Sera from infected animals reacted positively (1:625) with E. macaci antigen, but not to Babesia bigemina or B. bovis antigen at 1:125 titer. CONCLUSION: This study has revealed high prevalence of E. macaci infection in all three widely distributed Kenyan non-human primates. With the continued use of these animals as models for human parasitic diseases, the presence of this highly enzootic parasite should be noted. PMID- 18507705 TI - The four mammalian splice variants encoded by the p21-activated kinase 3 gene have different biological properties. AB - The p21-activated kinases (PAK1), PAK2, and PAK3 are members of the PAK group I and share high sequence identity and common biochemical properties. PAK3 is specifically implicated in neuronal plasticity and also regulates cell cycle progression, neuronal migration, and apoptosis. Loss of function of PAK3 is responsible for X-linked non-syndromic mental retardation whereas gain of PAK3 function is associated with cancer. To understand the functional specificities of PAK3, we analyzed the structure of PAK3 gene products. We report here the characterization of a new alternatively spliced exon called c located upstream of the previously identified exon b. Exon b is detected in all tetrapods and not in fish, exon c is only present in mammals. Mammalian PAK3 genes encode four splice variants and the corresponding proteins were detected with specific antibodies in brain extracts. All PAK3 transcripts are specifically expressed in brain and in particular in neurons. The presence of the exons b and c renders the kinase constitutively active and decreases interaction with GTPases. The expression of the new splice variants in COS7 cells alters cell morphology and modifies the structure of focal adhesions. We propose that the appearance of new alternatively spliced exons during evolution and the resulting increase of complexity of PAK3 gene products may confer new functions to this kinase and contribute to its specific roles in neuronal signaling. PMID- 18507706 TI - Toward disentangling sources of individual differences in appraisal and anger. AB - A theoretical framework is presented to explain individual differences in situation-specific emotional experience in terms of three different sources of variance: (a) individual differences in how one appraises one's circumstances, (b) individual differences in how appraisals are related to the experience of emotion, and (c) individual differences independent from situation and appraisal. The relative contribution and nature of these sources was examined empirically for the experience of anger based on data from two directed imagery studies (total N=1,192). Consistent results across the two studies demonstrated that variability in anger experience primarily stems from variability in how a situation is appraised and to a smaller extent from individual differences in the relations between the appraisals and anger and individual differences independent of appraisal. The findings further identified frustration as the central appraisal involved in anger. Implications for emotion theories and anger management programs are discussed. PMID- 18507707 TI - What underlies appraisals? Experimentally testing a Knowledge-and-Appraisal Model of Personality Architecture among smokers contemplating high-risk situations. AB - We tested a theoretical model of personality structures underlying patterns of intra-individual variability in contextualized appraisals. The KAPA (Knowledge and-Appraisal Personality Architecture) model was tested experimentally among smokers appraising their efficacy to resist the urge to smoke in high-risk situations. In a novel design, we assessed self-knowledge and situational beliefs idiographically and employed cognitive priming to manipulate the accessibility of self-knowledge experimentally. The results confirmed the unique KAPA-model prediction that priming would affect appraisals in a contextualized manner. Priming positively valenced self-knowledge enhanced self-efficacy appraisals specifically within that subset of situations that were relevant to the primed knowledge. The results were consistent with the hypothesis that systems of self- and situational knowledge underlie consistency and variability in appraisals. PMID- 18507710 TI - Egos inflating over time: a cross-temporal meta-analysis of the Narcissistic Personality Inventory. AB - A cross-temporal meta-analysis found that narcissism levels have risen over the generations in 85 samples of American college students who completed the 40-item forced-choice Narcissistic Personality Inventory (NPI) between 1979 and 2006 (total n=16,475). Mean narcissism scores were significantly correlated with year of data collection when weighted by sample size (beta=.53, p<.001). Since 1982, NPI scores have increased 0.33 standard deviation. Thus, almost two-thirds of recent college students are above the mean 1979-1985 narcissism score, a 30% increase. The results complement previous studies finding increases in other individualistic traits such as assertiveness, agency, self-esteem, and extraversion. PMID- 18507711 TI - An inductive exploration of the social effectiveness construct space. AB - There is no agreement regarding the nature or number of dimensions that make up the social effectiveness domain. We inductively explore the relationships between a set of social effectiveness measures with the intention of identifying an initial set of dimensions. An exploratory factor analysis of the Social Competence Inventory (SCI, Schneider, 2001) resulted in the identification of four factors: Social Potency, Social Appropriateness, Social Emotional Expression, and Social Reputation. A joint factor analysis between the SCI and a set of extant measures resulted in the identification of the same four factors. A fifth factor emerged when a set of scales from an emotional intelligence measure was included in the analysis, suggesting that emotional intelligence is not captured within the common factor space defined by measures of social effectiveness. This study represents a first step in the establishment of a set of common social effectiveness dimensions. PMID- 18507712 TI - Protection against cell death and sustained tyrosine hydroxylase phosphorylation in hydrogen peroxide- and MPP-treated human neuroblastoma cells with melatonin. AB - Neuroprotective effects of melatonin against oxidative stress-induced neuronal cell degeneration in human SH-SY5Y neuroblastoma cells were investigated in this report. The results demonstrate that exogenous administration of H(2)O(2) and 1 methyl, 4-phenyl, pyridinium ion (MPP(+)) significantly decreased cell viability in SH-SY5Y cultured cells. Desipramine, a monoamine uptake blocker was able to abolish the toxic effects of MPP(+) but not H(2)O(2) in reduction of cell viability. Conversely, melatonin reversed the toxic effects of H(2)O(2) and MPP(+) on cell viability. In addition, the reduction of phosphorylation of tyrosine hydroxylase, the rate limiting enzyme in dopamine synthesis, and phosphorylation of cyclic AMP responsive element-binding protein by H(2)O(2) and MPP(+) was also diminished by melatonin. These results demonstrate some effective roles of melatonin on neuroprotection and its action on the modulation of tyrosine hydroxylase phosphorylation. PMID- 18507713 TI - Treatment with testosterone or estradiol in melatonin treated females and males MRL/MpJ-Faslpr mice induces negative effects in developing systemic lupus erythematosus. AB - MRL/MpJ-Fas(lpr) mice is widely accepted as a valuable model of systemic lupus erythematosus. As described in a previous work, the incidence of lupus in this strain is determined by sex hormones, i.e., estrogens and androgens. Moreover, we reported that the immunomodulatory action of melatonin in these mice was gender dependent probably through modulation and inhibition of sex hormones. Herein, we performed an experiment using hormone therapy, by treating female MRL-lpr mice with testosterone and males with estradiol and with melatonin. A decrease in total serum immunoglobulin (Ig)G and IgM immunoglobulin titers, anti-double stranded DNA, and anti-CII autoantibodies in female mice treated with both melatonin and testosterone was revealed, along with an increase in pro inflammatory cytokines [interleukin (IL)-2, IL-6, interferon-gamma, tumor necrosis factor-alpha, and IL-1beta), nitrite/nitrate and a decrease in anti inflammatory cytokines (IL-10). Melatonin and estradiol treatment exhibited a similar effect in male mice. Autoantibody titer elevation and pro-inflammatory versus anti-inflammatory cytokine prevalence degraded all immunological parameters. Similar results were obtained when spleen and lymph node lymphocytes were cultured. Again, melatonin and testosterone treatment stimulated pro inflammatory and reduced anti-inflammatory cytokines produced by lymphocytes in females. The effect was similar in males treated with melatonin and estradiol. In summary, we observed that although melatonin alone prevents lupus development in females, adding testosterone, increased pro-inflammatory cytokine pattern. In contrary, estradiol-treated males did not show any decrease in pro-inflammatory cytokines but showed an increase in regard to melatonin controls. These findings confirm that melatonin action in MRL/MpJ-Fas(lpr) mice could be gender-dependent through modulation of sex hormones. PMID- 18507714 TI - Estrogen cross-talk with the melatonin signaling pathway in human osteoblasts derived from adolescent idiopathic scoliosis patients. AB - Adolescent idiopathic scoliosis (AIS) represents the most frequently occurring form of scoliosis that occurs and progresses in puberty. This critical period coincides with many biological changes related to estrogens. The aim of this study was to determine the effect of 17-beta-estradiol on the responsiveness of AIS osteoblasts to melatonin and the cross-talk between estrogen and melatonin at the levels of the G(S)alpha and G(i)alpha proteins. Human osteoblasts derived from AIS (n = 40) and control patients (n = 10) were first screened for their functional response to the melatonin and 17-beta-estradiol. In response to the 17 beta-estradiol in a specific group of scoliotic patients, the level of 3',5' cyclic adenosine monophosphate (cAMP) was significantly decreased when compared with the level observed in the presence of increasing concentrations of melatonin alone. Ours results provide strong evidence of the cross-talk between 17-beta estradiol and melatonin signaling in human AIS osteoblasts. These results indicate a novel role for 17-beta-estradiol and melatonin in AIS, controlling the coupling of G(S)alpha protein and MT2 receptor on human osteoblasts. We found that the increased cAMP levels induced by melatonin can be corrected by the treatment of the cells with 17-beta-estradiol. Thus, estrogens or estrogen receptor agonists become important compounds to consider in AIS osteoblast cell functioning. Consequently, our results add a new facet to the understanding the role and function of melatonin in AIS. PMID- 18507715 TI - Delayed surgical repair of penile fracture under local anesthesia. AB - INTRODUCTION: Penile fracture is a traumatic rupture of the tunica albuginea because of blunt injury of an erect penis. AIM: To assess the efficacy of a simple delayed surgical repair of penile fracture after a conservative treatment under local anesthesia in patients presented after 24 hours. METHODS: Twenty-four patients with penile fracture presented after 24 hours were subjected to history taking, clinical examination, urine analysis, and penile ultrasound. They underwent conservative treatment for 7-12 days, and then a surgical repair under local anesthesia was carried out. MAIN OUTCOME MEASURES: A follow-up for 6 months for sexual activity and any associated complaints in addition to local examination. RESULTS: All cases were presented with unilateral single tear, and the main cause of penile fracture was sexual intercourse. No intraoperative or postoperative complications were encountered. They regained their sexual activity 4-6 weeks after the repair. One case developed a mild penile deviation that did not interfere with sexual relation after the 6-month follow-up. CONCLUSION: Surgical repair of penile fracture after a conservative treatment is an effective method for patients with delayed presentation devoid of urethral involvement. PMID- 18507716 TI - Clinical trial methodology in premature ejaculation observational, interventional, and treatment preference studies--part II--study design, outcome measures, data analysis, and reporting. AB - INTRODUCTION: Regulatory approval of new drug treatments for premature ejaculation (PE) demands evaluation in well-designed clinical efficacy and safety randomized clinical trials (RCTs). AIMS: The objective of this article was to make recommendations for trial design and efficacy outcome measures which comprise ideal PE observational, interventional, and treatment preference trial methodology. MAIN OUTCOME MEASURES: Published data on clinical trial design, epidemiology, definitions, dimensions, and psychological impact of PE. METHODS: Data on the epidemiology, definitions, dimensions, and psychological impact of PE were reviewed, critiqued, and incorporated into a series of recommendations for standardization of PE clinical trial design, outcome measures, and reporting using the principles of evidence-based medicine. RESULTS: PE observational trials should be prospective and should provide quantitative or qualitative data derived from objective outcome measurements and/or the results of subject interview and other trial-specific investigations for analysis. PE drug trials should employ a double-blind RCT methodology and should include placebo control, active standard drug control, and/or dose comparison trials. Application of the placebo concept to psychotherapy intervention trials is complex and fraught with both conceptual and pragmatic problems. Criteria for the ideal PE preference trial are not yet determined but are likely to be a double-blind, crossover RCT of treatment-naive subjects using randomized drug sequences of equivalent drug doses. Ejaculatory latency time (ELT) and subject/partner outcome measures of control, personal/partner/relationship distress, and other study-specific outcome measures should be used as outcome measures. There is currently no published literature which identifies a clinically significant threshold response to intervention. CONCLUSION: Data from PE observational, interventional, and preference studies are only reliable, interpretable, and capable of being generalized to patients with PE when derived from well-designed observational studies or intervention RCTs using ELT and subject/partner-reported outcome measures of perceived ejaculatory control and personal/partner/relationship distress are used as trial outcome measures. PMID- 18507717 TI - Clinical trial methodology in premature ejaculation observational, interventional, and treatment preference studies--part I--defining and selecting the study population. AB - INTRODUCTION: Large, well-designed observational or clinical efficacy and safety randomized clinical trials are required to identify the prevalence of premature ejaculation (PE) and its associated risk factors, to characterize the dimensions of PE and the basis for treatment-seeking behaviour, and to achieve regulatory approval of new drug treatments. AIMS: The objective of this article was to make recommendations for the criteria for defining and selecting the study population. MAIN OUTCOME MEASURES: Contemporary published data on clinical trial design and the epidemiology, definitions, dimensions, and psychological impact of PE. METHODS: Contemporary data on the epidemiology, definitions, dimensions, and psychological impact of PE were reviewed, critiqued using the principles of evidence-based medicine, and incorporated into a series of evidence-based recommendations for standardization of patient selection for clinical trials in PE. RESULTS: Data from PE observational, interventional, and treatment preference studies are only reliable, interpretable, and capable of being generalized to patients with PE when study populations are defined by the constructs of an ejaculatory latency time of less than about 1 minute on all or nearly all occasions, the inability to delay ejaculation, and the presence of negative personal consequences, such as distress, bother, frustration, and/or the avoidance of sexual intimacy. CONCLUSION: These constructs can be incorporated into a multidimensional evidence-based definition of PE and/or single-item questions or multi-item diagnostic questionnaires. The International Society of Sexual Medicine definition of PE reflects the contemporary understanding of PE, represents the state-of-the-art multidimensional definition of PE, and is recommended as the basis of diagnosis of PE for all PE clinical trials. PMID- 18507718 TI - A mindfulness-based group psychoeducational intervention targeting sexual arousal disorder in women. AB - INTRODUCTION: Despite their widespread prevalence, there are no existing evidence based psychological treatments for women with sexual desire and arousal disorder. Mindfulness, the practice of relaxed wakefulness, is an ancient eastern practice with roots in Buddhist meditation which has been found to be an effective component of psychological treatments for numerous psychiatric and medical illnesses. In recent years, mindfulness has been incorporated into sex therapy and has been found effective for genital arousal disorder among women with acquired sexual complaints secondary to gynecologic cancer. AIM: The aim of this study was to adapt an existing mindfulness-based psychoeducation (PED) to a group format for women with sexual desire/interest disorder and/or sexual arousal disorders unrelated to cancer. METHODS: Twenty-six women participated in three 90 minute sessions, spaced 2 weeks apart, with four to six other women. Group PED was administered by one mental health trained provider and one gynecologist with post graduate training and experience in sexual medicine. MAIN OUTCOME MEASURES: Prior to and following the group, women viewed audiovisual erotic stimuli and had both physiological (vaginal pulse amplitude) and subjective sexual arousal assessed. Additionally, they completed self-report questionnaires of sexual response, sexual distress, mood, and relationship satisfaction. RESULTS: There was a significant beneficial effect of the group PED on sexual desire and sexual distress. Also, we found a positive effect on self-assessed genital wetness despite little or no change in actual physiological arousal, and a marginally significant improvement in subjective and self-reported physical arousal during an erotic stimulus. A follow-up comparison of women with and without a sexual abuse history revealed that women with a sexual abuse history improved significantly more than those without such history on mental sexual excitement, genital tingling/throbbing, arousal, overall sexual function, sexual distress, and on negative affect while viewing the erotic film. Moreover, there was a trend for greater improvement on depression scores among those with a sexual abuse history. CONCLUSIONS: These data provide preliminary support for a brief, three session group psychoeducational intervention for women with sexual desire and arousal complaints. Specifically, women with a history of sexual abuse improved more than women without such a history. Participant feedback indicated that mindfulness was the most effective component of the treatment, in line with prior findings. However, future compartmentalization trials are necessary in order to conclude this more definitively. PMID- 18507719 TI - Endoscopic retrieval of a penile prosthesis' rear-tip extender. AB - INTRODUCTION: The renewal of penile prosthesis is a foreseeable intervention because of the limited survival time of the different implants. It usually becomes necessary simply for material fatigue after long-term regular use; in some cases, premature intervention might be necessary because of infection or other perioperative complications. AIM: In the case presented here, the removal of a Dynaflex penile prosthesis showed a missing rear-tip extender on the right hand tube. MAIN OUTCOME MEASURES AND METHODS: A cavernoscopy of the right cavernous body was performed by using a conventional urethrocystoscope. The found rear-tip extender was extracted by using alligator forceps via the cystoscopic device. RESULTS: A new AMS CX hydraulic cavernous body prosthesis was successfully implanted. It has proven fully functional up to the current follow up control. CONCLUSIONS: This case shows that cavernoscopy is a gentle intraoperative procedure for diagnosing and removing material in penile prosthesis surgery. PMID- 18507720 TI - Penile constriction devices: case report, review of the literature, and recommendations for extrication. AB - INTRODUCTION: Penile constriction devices often present significant challenges to urologic surgeons. Failure to remove such devices can lead to significant ischemia and loss of tissue. Patients often present after several days of ischemia and swelling have developed. AIM: This article reviews previously published data on penile constriction devices and strategies for their removal. Additionally, we present new methodologies for extrication. METHODS: A comprehensive review of the English language literature was performed using MEDLINE. "Penile incarceration" and "penile strangulation" were used as search terms, and a manual bibliographic review of cross-referenced items was performed. Publications prior to 1970 were excluded from our search. MAIN OUTCOME MEASURES: Review of published literature on penile constriction devices and their removal. RESULTS: Penile incarceration is frequently described in the literature as an isolated case report or small series describing the approach of a single physician or group of physicians for dealing with these problems. Penile incarceration has been reported in a wide spectrum of age groups, with the incarcerating object most frequently placed for erotic or autoerotic purposes. While the most commonly reported devices causing incarceration are metal rings, higher-grade penile injuries are more frequently sustained by nonmetallic objects. Patients who present with incarceration after 72 hours are more likely to sustain higher-grade injuries than those who seek more timely treatment. Strategies for extrication depend on the type of device used, the length of time of incarceration, the patient's ability to remain calm, and the tools available to the presenting physicians. CONCLUSION: Penile incarceration is a urologic emergency with potentially severe clinical consequences. With rapid intervention and removal of the foreign body, most patients do extremely well and need no further intervention. Removal of such devices can be challenging and often requires resourcefulness and a multidisciplinary approach. PMID- 18507721 TI - Response of fractional synthesis rate (FSR) of fibrinogen, concentration of D dimer and fibrinolytic balance to physical activity-based intervention in obese children. AB - BACKGROUND: Physical activity-induced reduction in obesity-related hyperfibrinogenemia in children has been reported. The underlying mechanisms remain elusive. Further, the effect of such interventions on fibrinolysis in children is scarce. OBJECTIVES: To investigate in obese children, before and after a physical activity-based intervention: (i) the mechanistic role of fractional synthesis rate (FSR) of fibrinogen in the reduction of hyperfibrinogenemia; and (ii) the changes in fibrinolytic factors. METHODS: Subjects included 21 (age > 14 < 18 years; Tanner stage, IV-V) children (15 obese, BMI >95%tile for age and sex and six lean, BMI <85%tile). After baseline measurements of FSR of fibrinogen, and concentrations of fibrinogen, D-dimer, PAI 1 and t-PA in all children, studies were repeated after a 3-month randomized controlled physical activity-based lifestyle intervention in obese children only. RESULTS: FSR of fibrinogen was higher (P = 0.002) in the obese (vs. lean) group, which was reduced (P = 0.001) after intervention. This almost completely accounted for the reduction in obesity-related hyperfibrinogenemia. High levels of D-dimer decreased (P = 0.001) after intervention, whereas fibrinolysis was not enhanced. CONCLUSIONS: The direct reduction in the FSR of fibrinogen and the remarkable correlation between the magnitudes of reduction in fibrinogen FSR and concentration signify a mechanistic role for FSR in the regulation of physical activity-induced reversal of hyperfibrinogenemia in obese children. The congruent reductions in the FSR of fibrinogen and the concentrations of fibrinogen and D dimer in response to intervention despite depressed fibrinolysis suggest an overall improvement in the hypercoagulable state in obese children with physical activity-based lifestyle intervention. PMID- 18507722 TI - The quantitative detection of blurring of vision after eyedrop instillation using a functional visual acuity system. PMID- 18507723 TI - Cancellation of surgical day cases in an ophthalmic centre. PMID- 18507724 TI - Effects of repeated injection of intravitreal triamcinolone on macular oedema in central retinal vein occlusion. AB - PURPOSE: To investigate the effectiveness of repeated injections of intravitreal triamcinolone acetonide (IVTA) in the treatment of macular oedema caused by central retinal vein occlusion (CRVO). METHODS: Seventeen pseudophakic or aphakic eyes of 17 patients (10 male, seven female) with macular oedema caused by CRVO received a repeat injection of 4 mg IVTA, 16 weeks after the first injection of the same dose. The examination included measurements of best-corrected visual acuity (BCVA) for distance and central foveal thickness (CFT) by optical coherence tomography (OCT), preoperatively and 1, 2, 3 and 4 months postoperatively. The values were compared by paired-t test. Side-effects were monitored. RESULTS: BCVA and CFT were not significantly different before initial and repeat injections. Transient improvements of BCVA and CFT were achieved after both injections. At the end of follow-up, BCVA and CFT were significantly different compared to pre-injection values in the same group (P = 0.032, 0.049 in the initial-injection group and P = 0.001, 0.008 in the repeat-injection group, respectively). However, compared to the initial injection, BCVA measurements were significantly worse at each time-point (P = 0.043, 0.011, 0.010 and 0.012, respectively) after the repeat injection, as were CFT at 1, 2 and 3 months post injection (P = 0.040, 0.015 and 0.025, respectively). The achieved maximum mean intraocular pressures were 20.00 [standard deviation (SD) 2.06] mmHg and 18.56 (SD 3.65) mmHg after the first and repeat injections, respectively. These values were not significantly different (P = 0.467). No other significant adverse events were noted during the study. CONCLUSION: A repeat injection of 4 mg IVTA may not be as effective as an initial injection for the treatment of macular oedema caused by CRVO. PMID- 18507725 TI - Re-examination of organ-cultured, cryopreserved human corneal grafts after 27 years. AB - PURPOSE: To determine the long-term fate of cryopreserved corneas. Review of 17 organ-cultered cryopreserved corneas grafted in 1978-1979. METHODS: We measured visual acuity and refraction and performed biomicroscopy, applanation tonometry and optical pachometri (CCT). Endothelial photos were taken, cells were counted and morphology was studied. RESULTS: Four of 16 grafted corneas were still clear after 27 years. Mean CCT was 0.52 mm, endothelial cell density was 882 cells/mm(2) and visual acuity was 0.25 or better with an average of 0.6 in the four patients. Cell morphology showed irregularity in shape and size. CONCLUSION: This study shows that cryopreserved endothelium can function as well as non frozen corneas and that a regular hexagonal pattern is not essential for corneal clarity. The four grafts showed long-term durability despite the irregularity in shape and size. PMID- 18507726 TI - Systemic lupus erythematosus-associated optic neuritis: clinical experience and literature review. AB - PURPOSE: This study aimed to evaluate the visual outcome of optic neuritis in patients with systemic lupus erythematosus (SLE). METHODS: A retrospective, case observational study was conducted by reviewing eight patients with SLE-associated optic neuritis from January 1986 to October 2004. The demographic data, clinical manifestations, laboratory data, medical management and disease courses of these eight patients were retrospectively analysed. Main outcome measurements included final visual acuity (VA) and relapse of optic neuritis. Statistical analyses were made using the chi-square test and a linear regression model. The English language literature on SLE-associated optic neuritis was reviewed. RESULTS: Initial visual loss was severe in SLE-associated optic neuritis. Seven patients (87%) had VA < 20/200 at onset. All patients received steroid pulse therapy followed by oral steroid tapering. Final visual outcome was highly variable, ranging from the complete recovery of VA in four patients, to partial recovery in one and poor recovery in three. Better visual recovery occurred in patients who received earlier treatment (within 10 days). However, longer duration of steroid administration was found to have no significant benefit on visual outcome. CONCLUSIONS: Systemic lupus erythematosus-associated optic neuritis is not common. However, it is important that ophthalmologists differentiate SLE associated optic neuritis from idiopathic optic neuritis because of the severe visual impairment and steroid dependence associated with the former. Early diagnosis and prompt treatment are important for restoring visual function in these patients. PMID- 18507727 TI - Medical treatment for combined Fusarium and Acanthamoeba keratitis. AB - PURPOSE: Acanthamoeba and fungal keratitis are rare ocular infections. We report cases of combined Fusarium and Acanthamoeba keratitis and the clinical course of medical treatment. METHODS: We reviewed the medical records of patients treated for culture-proven Acanthamoeba keratitis at a referral centre, during 2001-2006. RESULTS: Eleven consecutive patients were treated for culture-proven Acanthamoeba keratitis during the 5 years, two of whom had combined fungal infections. A 29 year-old man presented with ground-glass corneal oedema and epitheliopathy caused by contact lens use. The other patient, a 7-year-old girl, had eye trauma that led to a feathery corneal infiltrate. Both cases were treated with topical 0.02% polyhexamethylene biguanide (PHMB), 0.1% propamidine, 1% clotrimazole and 5% natamycin. Therapeutic keratoplasty was not required in either case. CONCLUSIONS: Timely identification of the pathogen, with repeated culture and smear if necessary, as well as adequate dosage to prevent recurrence is highly recommended in order to preclude the need for therapeutic penetrating keratoplasty. PMID- 18507728 TI - Endothelial dysfunction in glaucoma. AB - Glaucoma is a group of ocular diseases characterized by optic neuropathy associated with loss of the retinal nerve fibre layer and re-modelling of the optic nerve head, and a subsequent particular pattern of visual field loss. Increased intraocular pressure is the most important risk factor for the disease, but the pathogenesis of glaucoma is not monofactorial. Among other factors, ischaemia and vascular dysregulation have been implicated in the mechanisms underlying glaucoma. The vascular endothelium plays an important role in the regulation of ocular blood flow and pathological alterations of vascular endothelial cells may induce ischaemia and dysregulation. The present review summarizes our current evidence of endothelial dysfunction in glaucoma. This is of interest because endothelial dysfunction is a good prognostic factor for progression in several diseases. Although such data are lacking for glaucoma, endothelial dysfunction may provide an attractive target for therapeutic intervention in open-angle glaucoma and other vascular disorders of the eye. PMID- 18507729 TI - Peribulbar fungal abscess and endophthalmitis following posterior subtenon injection of triamcinolone acetonide. PMID- 18507730 TI - Macular hole closure following intravitreal triamcinolone injection in a previously vitrectomized diabetic eye. PMID- 18507731 TI - Fundus autofluorescence related to retinal morphological and functional changes in idiopathic macular holes. AB - PURPOSE: To investigate fundus autofluorescence (FAF) characteristics in relation to morphological and functional features of idiopathic macular hole (IMH). METHODS: Twenty eyes of 20 consecutive patients with stage 3 or 4 macular holes were included. Fundus autofluorescence images were obtained using the Heidelberg retina angiograph 2, retinal structure was evaluated with 3-dimensional (3-D) imaging of Fourier-domain optical coherence tomography (FD-OCT), and retinal function was assessed with microperimetry-1 (MP-1). RESULTS: Markedly increased FAF in the foveal centre corresponding to the macular hole, confirmed with FD OCT, was demonstrated in all eyes. A surrounding hypoautofluorescent ring corresponded to the subretinal fluid cuff. The area of relatively reduced FAF around the ring corresponded precisely to retinal oedema. In 15 eyes (75%), a stellate appearance with dark radiating striae was seen in the relatively reduced FAF and was correlated with intraretinal cystic changes in the outer plexiform layer, observed by FD-OCT. Mean preoperative visual acuity was significantly poorer in eyes without a stellate appearance than in those with a stellate appearance (p = 0.023). The MP-1 study confirmed impaired retinal function in the macular hole bed and in the area of the fluid cuff and retinal oedema. CONCLUSIONS: Fundus autofluorescence imaging reflects anatomic changes and represents the dysfunctional retinal area in IMH. The technique provides 2-D images with 3-D information on the retinal morphology of this disease. PMID- 18507732 TI - Inflammatory choroidal neovascular membrane in presumed ocular Lyme borreliosis. AB - INTRODUCTION: Lyme disease is a multisystemic disease with protean ocular manifestations. We describe the occurrence of inflammatory choroidal neovascular membrane (CNVM) in two patients suffering from presumed Lyme disease. METHODS: Descriptive review of the clinical records of two patients. RESULTS: Patient 1: 16-year-old healthy male presenting with a visual acuity of counting fingers [oculus dexter (OD)] and 6/6 [oculus sinister (OS)] 3 months after a tick bite. He had papillitis and an exudative subretinal macular lesion OD. Treatment was started with intravenous (IV) ceftriaxone; a week later, IV methylprednisolone was administered with a tapering dose of oral steroids thereafter. Three months later, VA had improved to 3/60 OD. Patient 2: 38-year-old healthy female presenting with reduced left-eye vision (6/24) 6 weeks after a tick bite. She also suffered from erythema migrans and arthralgias. She had left-eye papillitis, macular haemorrhages and vascular sheathing. Treatment was started with IV ceftriaxone. One month later, there was profound loss of vision with development of CNVM. Treatment was declined by the patient and eventually retinal fibrosis developed. CONCLUSION: Inflammatory CNVM has not been described previously in the setting of ocular Lyme borreliosis. We herein describe the occurrence of inflammatory CNVM in two patients whose diagnosis with Lyme disease was clinically based--both were sero-negative. Visual outcome in the two patients was profoundly impaired because of the ensuing macular scar. PMID- 18507733 TI - A need for new diagnostic tools for giant cell arteritis. PMID- 18507734 TI - Will improvement of knowledge lead to improvement of compliance with glaucoma medication? AB - PURPOSE: To identify specific items on knowledge and need for information that could be used to improve compliance with glaucoma medication. METHODS: Forty-four randomly selected ophthalmologists assigned 166 consecutive glaucoma patients to complete a questionnaire. The questionnaire was developed systematically based on focus group interviews, consultation of experts and a pilot test. Items included knowledge about glaucoma and its treatment, need for information and compliance with therapy. RESULTS: No statistically significant correlation was found between the total level of knowledge and compliance. Concerning knowledge of glaucoma, one out of 19 items had a statistically significant negative correlation with compliance. Concerning knowledge of glaucoma treatment, one out of 18 items had a statistically significant positive correlation with compliance. Regarding total need for information, no statistically significant correlation with compliance was found. Three out of 22 items had a statistically significant positive correlation with compliance. CONCLUSION: Given the number of related items, their direction and limited strength of correlation and the difficulty of hypothesizing a causal relation for some items, it is unlikely that further improving knowledge will greatly improve compliance with glaucoma medication. Interventions focused on attitude and discipline may be of more benefit in improving compliance with therapy. PMID- 18507736 TI - Modulation of brain response to emotional images by alcohol cues in alcohol dependent patients. AB - Alcohol is often used to modulate mood states. Alcohol drinkers report that they use alcohol both to enhance positive affect and to reduce dysphoria, and alcohol dependent patients specifically state reduction of negative affect as a primary reason for drinking. The current study proposes that alcohol cues may reduce negative affect in alcoholics. We used functional magnetic resonance imaging to examine brain activation in response to combination images that juxtaposed negative or positive International Affective Picture System (IAPS) images with an alcohol or non-alcohol-containing beverage. We found that in the absence of the alcohol cue, alcoholics showed more activation to negative than to positive images and greater activation than controls to negative images. When the IAPS images were presented with the alcohol cue, there was a decreased difference in activation between the positive and negative images among the alcoholics, and a decreased difference in response to the negative images between controls and alcoholics. Additionally, in the neutral-beverage conditions, anxiety ratings significantly predicted activation in the right parahippocampal gyrus but did not predict activation when the alcohol cues were presented. In conclusion, the alcohol cues may have modulated cortical networks involved in the processing of emotional stimuli by eliciting a conditioned response in the alcoholics, but not in the controls, which may have decreased responsiveness to the negative images. PMID- 18507735 TI - Lack of effect of chronic dextromethorphan on experimental pain tolerance in methadone-maintained patients. AB - Good evidence exists to suggest that individuals on opioid maintenance for the treatment of addiction (i.e. methadone) are less tolerant of experimental pain than are matched controls or ex-opioid addicts, a phenomenon theorized to reflect opioid-induced hyperalgesia (OIH). Agonist activity at the excitatory ionotropic N-methyl-D-aspartate (NMDA) receptor on dorsal horn neurons has been implicated in the development of both OIH and its putative expression at the clinical level opioid tolerance. The aim of this study was to evaluate the potential utility of the NMDA-receptor antagonist, dextromethorphan (DEX), to reverse or treat OIH in methadone-maintenance (MM) patients. Utilizing a clinical trial design and double blind conditions, changes in pain threshold and tolerance [cold pressor (CP) and electrical stimulation (ES)] following a 5-week trial of DEX (titrated to 480 mg/day) in comparison with placebo was evaluated in a well-characterized sample of MM patients. The sample (n = 40) was 53% male and ethnically diverse (53% Latino, 28% African American, 10% White, 9% other), with a mean age of 48.0 years (SD = 6.97). Based on t-test analyses, no difference was found between groups on CP pain threshold, CP pain tolerance, ES pain threshold or ES pain tolerance, both pre- and postmedication. Notably, DEX-related changes significantly differed by gender, with women tending to show diminished tolerance for pain with DEX therapy. These results support that chronic high-dose NMDA antagonism does not improve tolerance for pain in MM patients, although a gender effect on DEX response is suggested. PMID- 18507738 TI - Myristoylation of p39 and p35 is a determinant of cytoplasmic or nuclear localization of active cyclin-dependent kinase 5 complexes. AB - Cdk5 is a member of the cyclin-dependent kinases (Cdks), activated by the neuron specific activator p39 or p35. The activators also determine the cytoplasmic distribution of active Cdk5, but the mechanism is not yet known. In particular, little is known for p39. p39 and p35 contain localization motifs, such as a second Gly for myristoylation and Lys clusters in the N-terminal p10 region. Using mutant constructs, we investigated the cellular distribution mechanism. We observed that p39 localizes the active Cdk5 complex in the perinuclear region and at the plasma membrane as does p35. We demonstrated the myristoylation of both p39 and p35, and found that it is a major determinant of their membrane association. Plasma membrane targeting depends on the amino acid sequence containing the Lys-cluster in the N-terminal p10 region. In contrast, a non myristoylated Ala mutant (p39G2A or p35G2A) showed nuclear localization with stronger accumulation of p39G2A than p35G2A. These results indicate that myristoylation regulates the membrane association of p39 as well as p35 and that the Lys cluster controls their trafficking to the plasma membrane. The differential nuclear accumulation of p39 and p35 suggests their segregated functions, p35-Cdk5 in the cytoplasm and p39-Cdk5 in the nucleus. PMID- 18507737 TI - Endoplasmic reticulum Ca2+ dysregulation and endoplasmic reticulum stress following in vitro neuronal ischemia: role of Na+-K+-Cl- cotransporter. AB - We investigated the role of Na(+)-K(+)-Cl(-) cotransporter (NKCC1) in conjunction with Na(+)/Ca(2+) exchanger (NCX) in disruption of endoplasmic reticulum (ER) Ca(2+) homeostasis and ER stress development in primary cortical neurons following in vitro ischemia. Oxygen-glucose deprivation (OGD) and reoxygenation (REOX) caused a rise in [Na(+)](cyt) which was accompanied by an elevation in [Ca(2+)](cyt). Inhibition of NKCC1 with its potent inhibitor bumetanide abolished the OGD/REOX-induced rise in [Na(+)](cyt) and [Ca(2+)](cyt). Moreover, OGD significantly increased Ca(2+)(ER) accumulation. Following REOX, a biphasic change in Ca(2+)(ER) occurred with an initial release of Ca(2+)(ER) which was sensitive to inositol 1,4,5-trisphosphate receptor (IP(3)R) inhibition and a subsequent refilling of Ca(2+)(ER) stores. Inhibition of NKCC1 activity with its inhibitor or genetic ablation prevented the release of Ca(2+)(ER). A similar result was obtained with inhibition of reversed mode operation of NCX (NCX(rev)). OGD/REOX also triggered a transient increase of glucose regulated protein 78 (GRP78), phospho-form of the alpha subunit of eukaryotic initiation factor 2 (p eIF2alpha), and cleaved caspase 12 proteins. Pre-treatment of neurons with NKCC1 inhibitor bumetanide inhibited upregulation of GRP78 and attenuated the level of cleaved caspase 12 and p-eIF2alpha. Inhibition of NKCC1 reduced cytochrome C release and neuronal death. Taken together, these results suggest that NKCC1 and NCX(rev) may be involved in ischemic cell damage in part via disrupting ER Ca(2+) homeostasis and ER function. PMID- 18507739 TI - Effects of chloramphenicol on brain energy metabolism using 31P spectroscopy: influences on sleep-wake states in rat. AB - Effects of chloramphenicol (antibiotic inhibiting complex-1 of respiratory chain) and thioamphenicol (TAP, a structural analog of CAP inactive on complex-1) were examined on cerebral energy metabolites and sleep-wake cycle architecture in rat. In the first group, animals were chronically equipped with a cranial surface resonator and (31)P spectroscopic measurements were performed using a 2 T magnetic resonance spectrometer (operating frequency 34.46 MHz). CAP administration (400 mg/kg, tail vein, light period) induced deficits in phosphocreatine (-30%, p < 0.01) and ATP (-40%, p < 0.01), whereas TAP (400 mg/kg) had no effect. In the second group, animals were chronically implanted with polygraphic electrodes for EEG and electromyogram recordings. CAP administered intraperitoneally at light-onset reduced rapid-eye movement (REM) sleep (-60% in the first 6 h of light period, p < 0.01), increased waking state (+65% in the first 6 h of light period, p < 0.01), and slightly affected slow wave sleep (SWS). During waking state, theta and sigma power bands of the EEG were, respectively, increased and decreased (p < 0.05). During SWS, delta power band was reinforced (p < 0.05), while theta, alpha, and sigma bands were decreased (p < 0.05). No changes occurred during REM sleep. TAP had no effect on sleep-wake states and spectral components of the EEG. Overall, these data indicate that REM sleep occurrence is linked to an aerobic production of ATP. PMID- 18507740 TI - Selfish genetic elements favor the evolution of a distinction between soma and germline. AB - Many multicellular organisms have evolved a dedicated germline. This can benefit the whole organism, but its advantages to genetic parasites have not been explored. Here I model the evolutionary success of a selfish element, such as a transposable element or endosymbiont, which is capable of creating or strengthening a germline-soma distinction in a primitively multicellular host, and find that it will always benefit the element to do so. Genes causing germline sequestration can therefore spread in a population even if germline sequestration is maladaptive for the host organism. Costly selfish elements are expected to survive only in sexual populations, so sexual species may experience an additional push toward germline-soma distinction, and hence toward cell differentiation and multicellularity. PMID- 18507741 TI - Evolutionary relationships among food habit, loss of flight, and reproductive traits: life-history evolution in the Silphinae (Coleoptera: Silphidae). AB - Flightlessness in insects is generally thought to have evolved due to changes in habitat environment or habitat isolation. Loss of flight may have changed reproductive traits in insects, but very few attempts have been made to assess evolutionary relationships between flight and reproductive traits in a group of related species. We elucidated the evolutionary history of flight loss and its relationship to evolution in food habit, relative reproductive investment, and egg size in the Silphinae (Coleoptera: Silphidae). Most flight-capable species in this group feed primarily on vertebrate carcasses, whereas flightless or flight dimorphic species feed primarily on soil invertebrates. Ancestral state reconstruction based on our newly constructed molecular phylogenetic tree implied that flight muscle degeneration occurred twice in association with food habit changes from necrophagy to predatory, suggesting that flight loss could evolve independently from changes in the environmental circumstances per se. We found that total egg production increased with flight loss. We also found that egg size increased with decreased egg number following food habit changes in the lineage leading to predaceous species, suggesting that selection for larger larvae intensified with the food habit change. This correlated evolution has shaped diverse life-history patterns among extant species of Silphinae. PMID- 18507742 TI - Sexual antagonism and the evolution of X chromosome inactivation. AB - In most female mammals, one of the two X chromosomes is inactivated early in embryogenesis. Expression of most genes on this chromosome is shut down, and the inactive state is maintained throughout life in all somatic cells. It is generally believed that X-inactivation evolved as a means of achieving equal gene expression in males and females (dosage compensation). Following degeneration of genes on the Y chromosome, gene expression on X chromosomes in males and females is upregulated. This results in closer to optimal gene expression in males, but deleterious overexpression in females. In response, selection is proposed to favor inactivation of one of the X chromosomes in females, restoring optimal gene expression. Here, we make a first attempt at shedding light on this intricate process from a population genetic perspective, elucidating the sexually antagonistic selective forces involved. We derive conditions for the process to work and analyze evolutionary stability of the system. The implications of our results are discussed in the light of empirical findings and a recently proposed alternative hypothesis for the evolution of X-inactivation. PMID- 18507743 TI - Rates and patterns in the evolution of snake-like body form in squamate reptiles: evidence for repeated re-evolution of lost digits and long-term persistence of intermediate body forms. AB - An important challenge in evolutionary biology is to understand how major changes in body form arise. The dramatic transition from a lizard-like to snake-like body form in squamate reptiles offers an exciting system for such research because this change is replicated dozens of times. Here, we use morphometric data for 258 species and a time-calibrated phylogeny to explore rates and patterns of body form evolution across squamates. We also demonstrate how time-calibrated phylogenies may be used to make inferences about the time frame over which major morphological transitions occur. Using the morphometric data, we find that the transition from lizard-like to snake-like body form involves concerted evolution of limb reduction, digit loss, and body elongation. These correlations are similar across squamate clades, despite very different ecologies and >180 million years (My) of divergence. Using the time-calibrated phylogeny and ancestral reconstructions, we find that the dramatic transition between these body forms can occur in 20 My or less, but that seemingly intermediate morphologies can also persist for tens of millions of years. Finally, although loss of digits is common, we find statistically significant support for at least six examples of the re-evolution of lost digits in the forelimb and hind limb. PMID- 18507744 TI - Transient SI and the dynamics of self-incompatibility alleles: a simulation model and empirical test. AB - A stochastic computer simulation model was created to compare the combined effects of selection and genetic drift on the dynamics of S-alleles under full sporophytic self-incompatibility (SI) versus transient SI, a form of partial SI in which flowers become self-compatible as they age. S-alleles were lost more rapidly with transient than with full SI, as is expected with weakened frequency dependent selection. Based on these results, equilibrium S-allele diversity is expected to be lower with partial SI for populations of comparable size and migration rates. Consistent with model results, a comparison of the proportion of incompatible crosses in full diallel experiments for a fully SI and a transiently SI species in the annual genus Leptosiphon suggests that S-allele diversity is lower in the partially SI species. Results of the simulation model indicate that the transmission advantage of self-fertilization can have complex effects on S allele dynamics in partial SI systems. PMID- 18507745 TI - Evolution on a local scale: developmental, functional, and genetic bases of divergence in bill form and associated changes in song structure between adjacent habitats. AB - Divergent selection on traits involved in both local adaptation and the production of mating signals can strongly facilitate population differentiation. Because of its links to foraging morphologies and cultural inheritance song of birds can contribute particularly strongly to maintenance of local adaptations. In two adjacent habitats--native Sonoran desert and urban areas--house finches (Carpodacus mexicanus) forage on seeds that are highly distinct in size and shell hardness and require different bite forces and bill morphologies. Here, we first document strong and habitat-specific natural selection on bill traits linked to bite force and find adaptive modifications of bite force and bill morphology and associated divergence in courtship song between the two habitats. Second, we investigate the developmental basis of this divergence and find that early ontogenetic tissue transformation in bill, but not skeletal traits, is accelerated in the urban population and that the mandibular primordia of the large-beaked urban finches express bone morphogenetic proteins (BMP) earlier and at higher level than those of the desert finches. Further, we show that despite being geographically adjacent, urban and desert populations are nevertheless genetically distinct corroborating findings of early developmental divergence between them. Taken together, these results suggest that divergent selection on function and development of traits involved in production of mating signals, in combination with localized learning of such signals, can be very effective at maintaining local adaptations, even at small spatial scales and in highly mobile animals. PMID- 18507746 TI - The use and abuse of surrogate endpoints in clinical research in transfusion medicine. PMID- 18507747 TI - Staff attitudes about event reporting and patient safety culture in hospital transfusion services. AB - BACKGROUND: Little is known about how transfusion service staff view issues pertaining to event reporting and patient safety. The goal of this study was to assess transfusion service staff attitudes about these issues. STUDY DESIGN AND METHODS: A survey was developed and administered to 945 transfusion service staff from 43 hospital transfusion services in the United States and 10 in Canada. The overall response rate was 73 percent (693 responses), with a mean of 15 respondents per site. RESULTS: While events resulting in patient harm are reported (91%) as well as mistakes not corrected that could cause harm (79%), less than one-third of respondents report deviations from procedures with no apparent potential to harm (31%) and mistakes that staff catch and correct on their own (27%). Staff indicated that the main reasons mistakes happen are interruptions (51%) and staff in other departments not knowing or understanding proper procedures (49%). Staff had overall positive attitudes about event reporting, but a significant minority were afraid of punitive consequences. Most were positive about their supervisor's safety actions and believed that their transfusion service tries to identify causes of mistakes. Only 31 percent, however, agreed that nursing staff would work with the transfusion service to reduce mistakes. CONCLUSION: Overall, the transfusion services had very positive attitudes about event reporting and safety culture. Transfusion services do well recording events that result in patient harm or have the potential for harm, but there is a need to increase reporting of deviations from procedures and mistakes that staff catch and correct on their own. In addition, there are a few areas of safety culture that warrant improvement, particularly the transfusion service's work relationship with nursing staff. The study provides useful descriptive information about how staff view event reporting and safety-related issues and identifies strengths and areas for improvement. PMID- 18507748 TI - A clinical study on the feasibility of autologous cord blood transfusion for anemia of prematurity. AB - BACKGROUND: The objective was to investigate the use of autologous red blood cells (RBCs) derived from umbilical cord blood (UCB), as an alternative for allogeneic transfusions in premature infants admitted to a tertiary neonatal center. STUDY DESIGN AND METHODS: UCB collection was performed at deliveries of less than 32 weeks of gestation and processed into autologous RBC products. Premature infants requiring a RBC transfusion were randomly assigned to an autologous or allogeneic product. The primary endpoint was an at least 50 percent reduction in allogeneic transfusion needs. RESULTS: Fifty-seven percent of the collections harvested enough volume (> or =15 mL) for processing. After being processed, autologous products (> or =10 mL/kg) were available for 36 percent of the total study population and for 27 percent of the transfused infants and could cover 58 percent (range, 25%-100%) of the transfusion needs within the 21-day product shelf life. Availability of autologous products depended most on the gestational age. Infants born between 24 and 28 weeks had the lowest availability (17%). All products, however, would be useful in view of their high (87%) transfusion needs. Availability was highest (48%) for the infants born between 28 and 30 weeks. For 42 percent of the infants with transfusion needs in this group, autologous products were available. For the infants born between 30 and 32 weeks, autologous products were available for 36 percent of the infants. Transfusion needs in this group were, however, much lower (19%) compared to the other gestational groups. CONCLUSION: Autologous RBCs derived from UCB could not replace 50 percent of allogeneic transfusions due to the low UCB volumes collected and subsequent low product availability. PMID- 18507749 TI - One single dose of 200 microg of antenatal RhIG halves the risk of anti-D immunization and hemolytic disease of the fetus and newborn in the next pregnancy. AB - BACKGROUND: The objective was the evaluation of the effect of the Dutch national routine antenatal RhIG (anti-D) immunization prevention (RAADP) program comprising one single dose of 200 microg (1000 IU) of RhIG in the 30th week of pregnancy, restricted to women without a living child. STUDY DESIGN AND METHODS: A nationwide historic control study was performed. All newly detected anti-D immunized para-1 in 1999, 2002, and 2004 were included and classified on the basis of received prophylaxis during the first pregnancy: antenatal and postnatal versus only postnatal RhIG. The numbers of D- parae-1 who delivered a D+ first child before the introduction (control group) or after the introduction (intervention group) of the RAADP were calculated from Vital Birth Statistics (8,700 and 12,000, respectively). RESULTS: Fifty-eight newly detected anti-D immunizations in the first trimester were observed in the control group and 39 in the intervention group, which resulted in a significant reduction of the prevalence of new anti-D immunizations from 0.67 percent (95% confidence interval [CI], 0.50%-0.84%) to 0.31 percent (95% CI, 0.21%-0.41%). No reduction was observed in anti-D immunizations newly detected at the 30th-week screening (0.25%). A nonsignificant risk reduction of the risk of severe hemolytic disease of the fetus and newborn (HDFN) was found (0.23% vs. 0.10%). The numbers needed to treat to prevent one anti-D-immunized pregnancy and one case of subsequent severe HDFN were 357 and 1255, respectively. CONCLUSIONS: RAADP of one single dose of 200 microg of RhIG in addition to postnatal RhIG (200 microg) halves the risk of anti-D immunization and subsequent severe HDFN. PMID- 18507750 TI - Visceral leishmaniasis after renal transplantation: report of 4 cases in northeastern Brazil. AB - Visceral leishmaniasis (VL) is a well recognized opportunistic infection in immunosuppressed patients, which may cause febrile illness. We describe 4 renal transplant patients with VL in an endemic area in Brazil and their response to therapy. In 3 cases the diagnosis was confirmed by bone marrow aspirate that revealed the presence of Leishmania. In 1 case the bone marrow aspirate was inconclusive and the diagnosis was made through spleen biopsy that showed the presence of the parasite. VL needs to be considered as a cause of febrile illness in transplanted patients living in endemic areas. PMID- 18507751 TI - Iatrogenic emphysematous pyelonephritis in a renal transplant patient. AB - Emphysematous pyelonephritis (EPN) is a rare condition that typically occurs in patients with diabetes mellitus, urinary tract obstruction, or immunosuppression such as solid organ transplant recipients. It has high mortality and frequently requires nephrectomy to achieve cure, although percutaneous drainage has been reported to be successful in some patients. We report a renal transplant recipient with underlying diabetes mellitus who developed iatrogenic EPN. The patient initially presented with dyspnea and was admitted for cardiac evaluation. There was no evidence of urinary tract infection at the time of admission. The patient developed high-grade fever 3 days after admission. Despite intravenous (IV) antibiotic therapy, the patient developed acute renal failure requiring hemodialysis. Studies revealed Klebsiella bacteremia and EPN. We believe that urinary tract infection was precipitated by urinary bladder catheterization performed on the day of admission. Despite 2 weeks of IV antibiotic therapy, infection persisted with progressive extension of gas into the perinephric space on repeat imaging. The patient underwent a transplant nephrectomy with subsequent clinical recovery. This case illustrates that antibiotics alone are often inadequate to cure and preserve renal function in EPN despite immediate therapy. Furthermore, this patient underscores the risk of serious infection precipitated by urinary bladder catheterization in immunocompromised patients. PMID- 18507752 TI - Infectious complications in living-donor liver transplant recipients: a 9-year single-center experience. AB - BACKGROUND: Infectious complications following living-donor liver transplantation (LDLT) remain a major cause of morbidity and mortality. We analyzed the frequency and type of infectious complications according to the post-transplantation period, and their risk factors with regard to morbidity and mortality. METHODS: We retrospectively analyzed 208 subjects who had undergone LDLT during a 9-year period. RESULTS: The rate of infection was 1.69 per patient during the study period. The predominant infections were intra-abdominal infections (37.6%), primary bacteremia (17.4%), and pneumonia (14.5%). Within the first post transplant month, 140 (39.9%) infections were detected, and catheter-related coagulase-negative staphylococci (44) were the most common infectious agents. During the 2-6-month post-transplant period, 109 infectious episodes occurred (31.1%), and Enterococcus sp. (n=16) related to biliary infection was the most frequent isolate. After the sixth month, 96 infectious episodes (29%) occurred, and biliary tract-related Escherichia coli (n=19) was the major causative organism. The overall mortality was 24.5% (51/208); 1-year survival rate was 88% (196/208). Post-transplant infection-related mortality was 52.9% (27/51). Biliary tract complications, such as biliary stenosis or leakage, significantly increased the mortality (P=0.01); however, reoperation (retransplantation or resurgery for biliary tract obstruction/leakage or to control bleeding) significantly reduced the mortality (P=0.01). CONCLUSIONS: Our data showed that early catheter removal would mainly aid in reducing infectious complications in the 1-month post transplantation period. Aggressive management, including reoperation, would lower the mortality in the LDLT recipients. PMID- 18507753 TI - Lower-than-standard dose peg-IFN alfa-2a for chronic hepatitis C caused by genotype 2 and 3 is sufficient when given in combination with weight-based ribavirin. AB - Mono-therapy with pegylated interferon (peg-IFN) has shown that a lower-than standard dose yields the same sustained viral response (SVR) rates as standard doses for chronic hepatitis C virus (HCV) infection caused by genotypes 2 or 3. Our aim was to see if a fixed, lower-than-standard dose of peg-IFN alfa-2a (135 microg weekly) in combination with ribavirin 11 mg/kg daily for 24 weeks yields sufficient SVR rates for genotypes 2 or 3. Hundred consecutive patients with a mean age of 44 years (range 20-69 years), 59 with genotype 3 and 41 with genotype 2, were studied. Rapid viral response (RVR) with HCV-RNA <15 IU/mL at treatment week 4 and SVR were calculated. RVR was achieved by 28/40 (70%) patients with genotype 2 and 41/58 (71%) with genotype 3. Significantly more genotype 2 patients with RVR achieved SVR 27/28 (96%) than genotype 2 patients who failed to achieve RVR, 8/12 (66%), P = 0.009. The corresponding figures for genotype 3 patients were 39/41 (95%) vs 11/17 (65%), respectively, P = 0.002. In total, SVR was achieved by 35/41 (85%) patients with genotype 2 and 51/59 (86%) patients with genotype 3, respectively. We found that 135 microg peg-IFN alfa-2a weekly was sufficient for treatment of genotype 2 and 3 chronic hepatitis C when combined with RBV dosed daily according to body weight. This combination yielded high SVR rates (85-86%) and may be cost-saving. PMID- 18507754 TI - Clinical significance of precore and core promoter mutations in genotype D hepatitis B-related chronic liver disease. AB - SUMMARY: The impact of mutations in the precore and basal core promoter (BCP) regions of the hepatitis B virus on the course of chronic liver disease is not well established. We sought to examine the relationship of these characteristics to the clinical expression of liver disease in patients infected with genotype D chronic hepatitis B (CHB). BCP and precore mutations in 110 patients with genotype D1 CHB were determined and correlated with clinical phenotype. Of 110 patients, 95 (86.5%) were HBeAg-negative. Compared with HBeAg-positive subjects, HBeAg-negative patients were over a decade older and had lower viral loads (3.70 +/- 0.98 vs 5.77 +/- 0.69 log copies/ml, P < 0.001). The double mutation A1762T G1764A was more prevalent in patients with advanced liver disease (AdLD) and was associated with higher alanine aminotransferase and viral load. After adjusting for age, there was a more than fourfold increase in the risk of AdLD with this mutation (OR = 4.4; 95% CI: 1.13-16.92, P < 0.03). Conversely, the G1757A substitution was associated with protection, being 90% less frequent among patients with AdLD (P = 0.001). The results indicate that in genotype D CHB, the presence of the A1762T-G1764A mutation was associated with more aggressive liver disease while the G1757A substitution was associated with protection from advanced disease. PMID- 18507755 TI - Two simultaneous hepatitis B virus epidemics among injecting drug users and men who have sex with men in Buenos Aires, Argentina: characterization of the first D/A recombinant from the American continent. AB - Previous studies have revealed that hepatitis B virus (HBV)/D and HBV/F predominate among blood donors from Buenos Aires, Argentina. In the present study, blood samples from two high-risk groups were analysed: 160 corresponding to street- and hospital-recruited injecting drug users [81.2% showing the 'anti hepatitis B core antigen (anti-HBc) only' serological pattern] and 20 to hepatitis B surface antigen (HBsAg)(+)/anti-HBc(+) men who have sex with men. HBV genotypes were assigned by polymerase chain reaction amplification followed by restriction fragment length polymorphism and confirmed by nucleotide sequencing of two different coding regions. HBV DNA was detected in 27 injecting drug users (16.9%, occult infection prevalence: 7.7%), and 14 men who have sex with men (70%). HBV/A prevailed among injecting drug users (81.8%) while HBV/F was predominant among men who have sex with men (57.1%). The high predominance of HBV/A among injecting drug users is in sharp contrast to its low prevalence among blood donors (P = 0.0006) and men who have sex with men (P = 0.0137). Interestingly, all HBV/A S gene sequences obtained from street-recruited injecting drug users encoded the rare serotype ayw1 and failed to cluster within any of the known A subgenotypes. Moreover, one of the HBV strains from a hospital recruited injecting drug user was fully sequenced and found to be the first completely characterized D/A recombinant genome from the American continent. Data suggest that two simultaneous and independent HBV epidemics took place in Buenos Aires: one spreading among injecting drug users and another one sexually transmitted among the homosexual and heterosexual population. PMID- 18507756 TI - Bone marrow-derived mesenchymal stem cells are capable of mediating hepatitis B virus infection in injured tissues. AB - We have previously showed that endothelial progenitor cells (EPCs) through uptake of hepatitis B virus (HBV) may play a critical role in mediating extrahepatic HBV diseases. However, it remains to be elucidated whether mesenchymal stem cells (MSCs) are capable of mediating HBV trans-infection into extrahepatic tissues. METHODS AND RESULTS: In this study, we showed that HBV antigens, HBV DNA and the viral particles were detected in MSCs after 3 days virus challenge. Neither HBV covalently closed circular DNA nor pregenomic RNA were detected in MSCs. Intravenously transplantation of HBV-exposed MSCs into myocardial infarction mouse model resulted in incorporation of HBV into injured heart and other damaged tissues. CONCLUSION: These results indicate that MSCs could serve as an additional extrahepatic virus reservoir, which may play a role at least in part in mediating HBV trans-infection into the injured tissues through the process of MSCs recruitment. PMID- 18507758 TI - Could the depression of obese patients suffering from chronic hepatitis C be temporarily improved? AB - Depression is an usual finding in patients suffering from chronic hepatitis C. Development of moderate to severe depressive symptoms occurs frequently during pegylated interferon/ribavirin treatment and is generally predicted by baseline depression scores. Furthermore, the obese patients have been found to be twice as likely to suffer from anxiety, impaired social interaction, and depression when compared with the no obese population. In order to evaluate the efficacy of a pharmacological treatment of depression, 68 obese patients with chronic hepatitis C, under or not antiviral therapy, were selected and enrolled into this open, controlled pilot study. Our population was divided in two groups: 'on Selective Serotonin Reuptake Inhibitors plus support', with individual titration of medication to adequate side-effects, including thirty seven patients, and 'on only support', involving thirty one patients. Both groups were well balanced for gender, age and antiviral treatment. The selected patients had, at entry, a Beck Depression Inventory score of 24.5 +/- 8.1 (mean +/- SD). Therapeutic successful outcomes (a decreased score of >or= 10 units compared to the baseline) were statistically more frequent in antidepressant drug-treated group (P = 0.005); they were well predicted by dose of Selective Serotonin Reuptake Inhibitors. Thirty five percent of patients were non-responder to Selective Serotonin Reuptake Inhibitors. The drug tolerability was good. Nearly twenty percent of patients were responder to only support. PMID- 18507757 TI - The inverse relationship between chronic HBV and HCV infections among injection drug users is associated with decades of age and drug use. AB - Infection with hepatitis C virus (HCV) may suppress co-infection with hepatitis B virus (HBV) during acute or chronic HBV infection. We examined relationships between HBV infection, HCV infection and other factors among injection drug users (IDUs) with antibodies to both viruses. Participants enrolled in a cross sectional study during 1998-2000 were considered to have been infected with HBV if they had core antibody, to be chronically infected if they had hepatitis B surface antigen (HBsAg), to have been infected with HCV if they had HCV antibody and to be chronically infected if they had HCV RNA. Among 1694 participants with antibody to both viruses, HBsAg prevalence decreased with increasing age among those positive for HCV RNA [from 4.55% in those 18-29 years to 1.03% in those >or=50 years old (P(trend) = 0.02)], but not among those who were negative for HCV RNA. Chronic HBV infection was less common overall among those with chronic HCV infection (odds ratio [OR], 0.25; P < 0.0001), but this inverse relationship was much stronger in the oldest (>50 years; OR = 0.15) than the youngest (18-29 years; OR = 0.81) participants (P(trend) = 0.03). Similar results were obtained when duration of injection drug use was substituted for age (P(trend) = 0.05). Among IDUs who have acquired both HBV and HCV, chronic HBV infection is much less common among those with chronic HCV infection, but this inverse relationship increases markedly with increasing years of age and injection drug use. Co infection with HCV may enhance the resolution of HBsAg during the chronic phases of these infections. PMID- 18507759 TI - Clinical and histological impact of previous hepatitis B virus infection in patients with chronic hepatitis C. AB - BACKGROUND: Recent reports suggest that hepatitis C virus (HCV) carriers with serological markers of prior hepatitis B virus (HBV) infection have more advanced liver fibrosis, irrespective of HBV-DNA detection. AIMS: We sought to assess the prevalence and impact of previous HBV infection in patients with HCV chronic infection. METHODS: This cross-sectional study included hepatitis B surface antigen- and human immunodeficiency virus-negative subjects with positive HCV RNA. All patients had prior parenteral exposure as the probable source of HCV infection. Serum samples were tested for HBV-DNA using a commercial assay. The METAVIR system was used for histological analysis. RESULTS: One-hundred and eleven patients were evaluated. Thirty-one out of 111 patients (28%) tested positive for antihepatitis B core antigen (anti-HBc). HBV-DNA was not detected in any sample. Anti-HBc-positive patients showed higher histological grading, staging and a higher fibrosis progression rate. By multivariate analysis, anti HBc-positivity was predictive of moderate to severe activity [odds ratio (OR)=3.532; P=0.032] and significant hepatic fibrosis (OR=3.364; P=0.017). After approximately 20 years of infection, advanced liver fibrosis (F3/F4) can be expected in 13% of anti-HBc-negative subjects who acquired HCV before the age of 30 and in 57% of those anti-HBc-positive patients who were infected by HCV after 30 years of age (P<0.001). CONCLUSION: Previous HBV infection is common among HCV carriers and may exert a negative impact on the natural history of HCV infection, independently of the presence of significant HBV replication. PMID- 18507760 TI - Activated protein C prevents hepatic ischaemia-reperfusion injury in rats. AB - BACKGROUND: Hepatic ischaemia-reperfusion injury (IRI) is a serious complication of liver surgery, especially extended hepatectomy and liver transplantation. Activated protein C (APC), a potent anticoagulant serine protease, has been shown to have cell-protective properties by virtue of its anti-inflammatory and anti apoptotic activities. METHODS: The present study was designed to examine the cytoprotective effects of APC in a 60-min warm-IRI rat model. RESULTS: Following a single intravenous injection of APC before reperfusion, APC exerted cytoprotective effects 4 h after reperfusion, as evidenced by: (i) decreased levels of transaminase and improved histological findings of IRI, (ii) reduced infiltration and activation of neutrophils, macrophages and T cells, (iii) reduced expression of tumour necrosis factor-alpha, (iv) reduced expression of P selectin and intracellular adhesion molecule-1, (v) inhibited coagulation and attenuated sinusoidal endothelial cell injury, (vi) improved hepatic microcirculation and (vii) decreased transferase-mediated dUTP nick end-labelling positive cells. These effects of APC were observed 4 h but not 24 h after reperfusion. However, multiple injections of APC after reperfusion significantly decreased the levels of transaminase and the activity of myeloperoxidase, and improved histological findings of IRI 24 h after reperfusion. CONCLUSION: These results suggest that APC is a promising therapeutic option for hepatic warm-IRI; however, multiple injections of APC are necessary to maintain its cell-protective action over the long term. PMID- 18507761 TI - Contribution of hepatic stellate cells and matrix metalloproteinase 9 in acute liver failure. AB - BACKGROUND/AIMS: Fulminant hepatitis or acute liver failure (ALF), initiated by viral infection or hepatic toxin, is a devastating medical complication without effective therapeutic treatment. In this study, we addressed the potential roles of hepatic stellate cells (HSCs) and their produced matrix metalloproteinases (MMPs) in development of ALF. METHODS: Mice were given lipopolysaccharide (LPS) and beta-galactosamine (GA) or carbon tetrachloride to create ALF and establish the association of IL-1, MMP-9, and caspase-3 in acute liver failure. RESULTS: In response to the hepatic toxin, IL-1 and MMP-9 were promptly induced within 1 hour, followed by caspase-3 activation at 2 hours, and dehiscence of sinusoids at 4 hours, and consequent lethality. In contrast, MMP-9 knockout mice were resistant to lethality and absent of caspase-3 activation, demonstrating an MMP-9 dependent activation of caspase in vivo. Further, IL-1-receptor knockout mice were resistant to lethality in MMP-9 dependent manner, indicating a causative relationship. Although many hepatic cells are capable to produce MMP-9 in vitro, HSCs were demonstrated here as the major hepatic cells to express MMP-9 in liver injury. To recapitulate the sinusoidal microenvironment we cultured primary HSCs in 3-dimensional ECM. In response to IL-1, massive MMP-9 was produced by the 3D culture concomitantly with degradation of type-IV collagen. CONCLUSIONS: Based on these evidences, we propose a novel model to highlight the initiation of acute liver failure: IL-1-induced MMPs by HSCs within the space of Disse and thereafter ECM degradation may provoke the collapse of sinusoids, leading parenchymal cell death and loss of liver functions. PMID- 18507763 TI - Needlestick injuries among German medical students: time to take a different approach? AB - CONTEXT: Medical students are at risk of occupational exposure to blood-borne viruses following needlestick injuries (NSIs) during medical school. The reporting of NSIs is an important step in the prevention of further injuries and in the initiation of early prophylaxis or treatment. The objective of this study was to describe the mechanisms whereby medical students experience occupational percutaneous blood exposure through NSIs and to discuss rational strategies for prevention. METHODS: Incidents of exposure to blood-borne pathogens among medical students at a large German university were analysed. Year 6 medical students completed a written survey immediately before the clinical part of their training began, describing incidents that had occurred during the previous 5 years. RESULTS: In our study, 58.8% (183/311) of participating medical students recalled at least one NSI that had occurred during their studies. Overall, 284 NSIs were reported via an anonymous questionnaire. DISCUSSION: Occupational exposure to blood is a common problem among medical students. Efforts are required to ensure greater awareness of the risks associated with blood-borne pathogens among German medical students. Proper training in percutaneous procedures and how to act in the event of injury should be given in order to reduce the number of injuries. PMID- 18507762 TI - Involvement of p38 mitogen-activated protein kinase pathway in honokiol-induced apoptosis in a human hepatoma cell line (hepG2). AB - BACKGROUND: Honokiol has been known to have antitumour activity. This study was conducted to evaluate the antiproliferative potential of honokiol against the hepG2 heptocellular cell line and its mechanism of action. METHODS: hepG2 cells were treated with honokiol of 0-40 microg/ml concentration. The cytotoxic effect of honokiol was determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. The apoptosis was evaluated by flow cytometry. Western blots were used to analyse the expression of various proteins (procaspase-9, procaspase-3, cleaved caspase-3, cytochrome c, Bcl-2, Bax, Bad, Bcl-X(L) and p38). RESULTS: Honokiol induced apoptosis with a decreased expression of procaspase-3 and -9 and an increased expression of active caspase 3. Exposure of hepG2 cells to honokiol resulted in the downregulation of Bcl-X(L) and Bcl-2 expression and the release of mitochondrial cytochrome c to the cytosol. In addition, honokiol activated the p38 mitogen-activated protein kinase (MAPK) pathway, and the inhibition of this pathway by SB203580 reduced honokiol induced apoptosis and activation of caspase-3. CONCLUSION: Honokiol induces apoptosis of hepG2 human hepatocellular carcinoma cells through activation of the p38 MAPK pathway, and, in turn, activation of caspase-3. PMID- 18507764 TI - The medical student interviewer. PMID- 18507765 TI - Can patients refuse to be treated by medical students? PMID- 18507766 TI - Medical students' experiences with medical errors: an analysis of medical student essays. AB - OBJECTIVE: This study aimed to examine medical students' experiences with medical errors. METHODS: In 2001 and 2002, 172 fourth-year medical students wrote an anonymous description of a significant medical error they had witnessed or committed during their clinical clerkships. The assignment represented part of a required medical ethics course. We analysed 147 of these essays using thematic content analysis. RESULTS: Many medical students made or observed significant errors. In either situation, some students experienced distress that seemingly went unaddressed. Furthermore, this distress was sometimes severe and persisted after the initial event. Some students also experienced considerable uncertainty as to whether an error had occurred and how to prevent future errors. Many errors may not have been disclosed to patients, and some students who desired to discuss or disclose errors were apparently discouraged from doing so by senior doctors. Some students criticised senior doctors who attempted to hide errors or avoid responsibility. By contrast, students who witnessed senior doctors take responsibility for errors and candidly disclose errors to patients appeared to recognise the importance of honesty and integrity and said they aspired to these standards. CONCLUSIONS: There are many missed opportunities to teach students how to respond to and learn from errors. Some faculty members and housestaff may at times respond to errors in ways that appear to contradict professional standards. Medical educators should increase exposure to exemplary responses to errors and help students to learn from and cope with errors. PMID- 18507767 TI - Long-term follow-up of a 10-month programme in curriculum development for medical educators: a cohort study. AB - CONTEXT: There is an ongoing need for curriculum development (CD) in medical education. However, only a minority of medical teaching institutions provide faculty development in CD. This study evaluates the long-term impact of a longitudinal programme in curriculum development. METHODS: We surveyed eight cohorts of participants (n = 64) and non-participants (n = 64) from 1988 to 1996 at baseline and at 6-13 years after completion of a 10-month, one half-day per week programme offered annually, which included a mentored CD project, workshops on CD steps, a final paper and a presentation. RESULTS: Fifty-eight participants (91%) and 50 non-participants (78%) returned completed follow-up surveys. In analyses, controlling for background characteristics and baseline self-rated proficiencies, participants were more likely than non-participants at follow-up to report having developed and implemented curricula in the past 5 years (65.5% versus 43.7%; odds ratio [OR] 2.41, 95% confidence interval [CI] 1.03-5.66), to report having performed needs assessment when planning a curriculum (86.1% versus 58.8%; OR 5.59, 95% CI 1.20-25.92), and to rate themselves highly in developing (OR 3.57, 95% CI 1.36-9.39), implementing (OR 3.04, 95% CI 1.16-7.93) and evaluating (OR 2.74, 95% CI 1.10-6.84) curricula. At follow-up, 86.2% of participants reported that the CD programme had made a moderate or great impact on their professional careers. Responses to an open-ended question on the impact confirmed continued involvement in CD work, confidence in CD skills, application of CD skills and knowledge beyond CD, improved time management, and lasting relationships formed because of the programme. CONCLUSIONS: Our results suggest that a longitudinal faculty development programme that engages and supports faculty in real CD work can have long-lasting impact. PMID- 18507768 TI - Further challenges in measuring communication skills: accounting for actor effects in standardised patient assessments. AB - CONTEXT: Subjective rating scales for communication skills may yield more personally meaningful responses than more standardised rating schemes. It is unclear, however, whether such evaluations may be overly biased by respondents' rating styles, which may lead to unreliable measurement of examinees' communication skills. METHODS: Our study involved 212 students from the classes of 2005 and 2006 at the University of Rochester School of Medicine and Dentistry. All students were rated by actors depicting standardised patients (SPs) on the same seven cases using the 19-item Rochester Communication Rating Scale (RCRS). Different students were assigned to different actors playing the same SP. We assessed the extent to which actors' personal rating styles influenced the scores they assigned to students. Main outcome measures were: between-actor variability in responses; the degree to which actors' response styles contribute to overall scores, and improvements in reliability achieved by standardising actors' ratings. RESULTS: There were statistically significant differences between actors in their mean assigned scores. Scores aggregated over 18 separate SP cases have an expected generalisability coefficient of 0.79. If raw RCRS scores are used, a total of 27 replications of the RCRS are required to achieve a Cronbach's alpha of 0.8; standardisation reduces this number to 18. CONCLUSIONS: Although actors are variable in their use of a standardised subjective scale of communication, such differences contribute to an acceptably small proportion of the total variance if scores are combined across a large number of cases. Reliability can be markedly improved by standardising scores across raters. PMID- 18507769 TI - A controlled study of the short- and long-term effects of a Train the Trainers course. AB - Objectives This study aimed to establish the longterm effects of a 3-day 'Training for Trainers' course (TTC) on doctors' knowledge, teaching behaviour and clinical learning climate. Methods The study was designed as an intervention study with pre-, post- and long-term measurements. The intervention group (I group) included 118 doctors from the departments of internal medicine and orthopaedic surgery at one university hospital. The control group (C-group) consisted of 125 doctors from the corresponding departments at another university hospital. Gains in knowledge about teaching skills were assessed by a written test. Teaching behaviour and learning climate were evaluated by questionnaires. Results In the I-group, 98.4% of doctors, both specialists and trainees, participated in a TTC. Response rates on the written test varied from 90% at baseline to 70% at 6 months after the intervention. Knowledge about teaching skills increased in the I-group by 25% after the TTC and was sustained at 6 months. Questionnaire response rates varied from 98.4% at baseline to 84.8% at 6 months. Post-course, the teaching behaviour of the I-group significantly changed and its learning climate improved compared with the C-group. Scores for use of feedback and supervision in the I-group increased from 4-5 to 6-7 (maximum score = 9). This was significantly higher than in the C-group. Conclusions A 3-day residential TTC has a significant impact in terms of gains of knowledge concerning teaching skills, teaching behaviour and learning climate after 6 months. The positive effects demonstrated in this study were rooted in both the specialists and trainees who attended the course. PMID- 18507770 TI - Expression of proinflammatory cytokines and its relationship with virus infection in the brain of macaques inoculated with macrophage-tropic simian immunodeficiency virus. AB - The pathogenesis of acquired immunodeficiency syndrome dementia complex (ADC) is still poorly understood. Many studies suggest that proinflammatory cytokines such as IL-1beta and TNF-alpha released by microglia/macrophages or astrocytes play a role in CNS injury. A microscopic finding of a microglial nodule with multinucleated giant cells (MNGCs) is a histopathologic hallmark of ADC and named HIV encephalitis. However, in vivo expression of these cytokines in this microenvironment of HIV encephalitis is not yet clarified. One of the main reasons is complexities of brain pathology in patients who have died from terminal AIDS. In this study, we infected two macaques with macrophage-tropic Simian immunodeficiency virus SIV239env/MERT and examined expression of TNF-alpha and IL-1beta in inflammatory lesions with MNGCs and its relation to virus infected cells using immunohistochemistry. One macaque showed typical inflammatory lesions with MNGCs in the frontal white matter. Small microglial nodules were also detected in the basal ganglia and the spinal cord. SIVenv positive cells were detected mainly in inflammatory lesions, and seemed to be microglia/macrophages and MNGCs based on their morphology. Expression of IL-1beta and TNF-alpha were detected in the inflammatory lesions with MNGCs, and these positive cells were found to be negative for SIVenv by double-labeling immunohistochemistry or immunohistochemistry of serial sections. There were a few TNF-alpha positive cells and almost no IL-1beta positive cells in the area other than inflammatory lesions. Another macaque showed scattered CD3+ cells and CD68+ cells in the perivascular regions of the white matter. SIVenv and TNF-alpha was demonstrated in a few perivascular macrophages. These findings indicate that virus-infected microglia/macrophages do not always express IL-1beta and TNF alpha, which suggests an indirect role of HIV-1-infected cells in cytokine mediated pathogenesis of ADC. Our macaque model for human ADC may be useful for better understanding of its pathogenesis. PMID- 18507771 TI - Sensitivity of leaf size and shape to climate within Acer rubrum and Quercus kelloggii. AB - * Variation in the size and shape (physiognomy) of leaves has long been correlated to climate, and paleobotanists have used these correlations to reconstruct paleo-climate. Most studies focus on site-level means of largely nonoverlapping species sets. The sensitivity of leaf shape to climate within species is poorly known, which limits our general understanding of leaf-climate relationships and the value of intraspecific patterns for paleoclimate reconstructions. * The leaf physiognomy of two species whose native North American ranges span large climatic gradients (Acer rubrum and Quercus kelloggii) was quantified and correlated to mean annual temperature (MAT). Quercus kelloggii was sampled across a wide elevation range, but A. rubrum was sampled in strictly lowland areas. * Within A. rubrum, leaf shape correlates with MAT in a manner that is largely consistent with previous site-level studies; leaves from cold climates are toothier and more highly dissected. By contrast, Q. kelloggii is largely insensitive to MAT; instead, windy conditions with ample plant-available water may explain the preponderance of small teeth at high elevation sites, independent of MAT. * This study highlights the strong correspondence between leaf form and climate within some species, and demonstrates that intraspecific patterns may contribute useful information towards reconstructing paleoclimate. PMID- 18507772 TI - A plastid protein crucial for Ca2+-regulated stomatal responses. AB - * Guard cell movements are regulated by environmental cues including, for example, elevations in extracellular Ca(2+) concentration. Here, the subcellular localization and physiological function of the Ca(2+)-sensing receptor (CAS) protein was investigated. * CAS protein localization was ascertained by microscopic analyses of green fluorescent protein (GFP) fusion proteins and biochemical fractionation assays. Comparative guard cell movement investigations were performed in wild-type and cas loss-of-function mutant lines of Arabidopsis thaliana. Cytoplasmic Ca(2+) dynamics were addressed in plants expressing the yellow cameleon reporter protein YC3.6. * This study identified CAS as a chloroplast-localized protein that is crucial for proper stomatal regulation in response to elevations of external Ca(2+). CAS fulfils this role through modulation of the cytoplasmic Ca(2+) concentration. * This work reveals a novel role of the chloroplast in cellular Ca(2+) signal transduction. PMID- 18507773 TI - Sex-independent transmission ratio distortion system responsible for reproductive barriers between Asian and African rice species. AB - * A sex-independent transmission ratio distortion (siTRD) system detected in the interspecific cross in rice was analyzed in order to understand its significance in reproductive barriers. The S(1) gene, derived from African rice Oryza glaberrima, induced preferential abortion of both male and female gametes possessing its allelic alternative (), from Asian rice O. sativa, only in the heterozygote. * The siTRD was characterized by resolving it into mTRD and fTRD occurring through male and female gametes, respectively, cytological analysis of gametophyte development, and mapping of the S(1) locus using near-isogenic lines. The allelic distribution of the S(1) locus in Asian and African rice species complexes was also analyzed. * The siTRD system involved at least two components affecting male and female gametogeneses, respectively, including a modifier(s) that enhances fTRD. The chromosomal location of the major component causing the mTRD was delimited within an approx. 40 kb region. The S(1) locus induced hybrid sterility in any pairwise combination between Asian and African rice species complexes. * The allelic state of the S(1) locus has diverged between Asian and African rice species complexes, suggesting that the TRD system has a significant role in the reproductive barriers in rice. PMID- 18507776 TI - Does fructan have a functional role in physiological traits? Investigation by quantitative trait locus mapping. AB - * The role of fructan in growth and drought-stress responses of perennial ryegrass (Lolium perenne) was investigated in an F(2) mapping family that segregates for carbohydrate metabolism. * A quantitative trait locus approach was used to compare the genetic control of traits. * Growth and drought-stress traits were extremely variable within the family. Most traits had high broad-sense heritability. Quantitative trait loci (QTL) were identified for most traits; the maximum number of QTL per trait was four. Between 11% and 75% of total phenotypic variation was explained. Few growth-trait QTL coincided with previously identified fructan QTL. A cluster of drought-trait QTL was close to two previously identified regions of the genome with tiller base fructan QTL in repulsion. * The high sugar parent contributed few alleles that increased 'reserve-driven' growth or performance during drought-stress. Correlation of growth and drought-stress traits with fructan content was low and increasing fructan content per se would not appear to improve drought resistance. Complex patterns of carbohydrate partitioning and metabolism within the cell may explain contradictory relationships between carbohydrate content and growth/stress resistance traits. PMID- 18507777 TI - Physical activity interventions in Hispanic American girls and women. AB - The purpose of this article was to review physical activity interventions done with Hispanic American girls and women that were published between 1994 and 2007, and suggest ways of enhancing these interventions. A total of 12 such interventions were found. Majority of the interventions focused on both physical activity and nutrition behaviours. Only half of the interventions were based on a behavioural theory. Social cognitive theory was the most popular theory, which was operationalized by four interventions. The interventions ranged from 3 weeks to 2 years in duration. The impact was not necessarily linked to the length of the intervention. The most popular physical activity that was promoted was walking, which was utilized by four interventions. Most of the interventions utilized a classroom format for imparting instruction in being physically active. All the interventions utilized individual-level behaviour change as an approach, and none tried to address broader policy and environmental-level changes. Process evaluation was done by very few interventions and must be done more systematically. In terms of the impact, half of the interventions were successful in influencing the outcomes. Recommendations for enhancing the effectiveness of physical activity interventions in Hispanic American girls and women are presented. PMID- 18507779 TI - On the true origins of the Walther's ganglion blockade and more. PMID- 18507781 TI - Regarding CYP450 2D6 poor metabolizers. PMID- 18507782 TI - Mast cells are activated by Leishmania mexicana LPG and regulate the disease outcome depending on the genetic background of the host. AB - The regulatory effect of mast cells on the pathogenesis of leishmaniasis is unclear. We report a comparative analysis of TLR2 membrane expression, TNF-alpha, IL-10 and MIP-1alpha production, and granule release of bone marrow-derived mast cells (BMMCs) from susceptible BALB/c and resistant C57BL/6 mice, stimulated in vitro with Leishmania mexicana lipophosphoglycan (LPG). We studied the kinetics of mast cell degranulation and parasite numbers in lesions of both mouse strains infected with L. mexicana. We found that BMMCs of C57BL/6 mice expressed more TLR2 and produced higher levels of both cytokines and MIP-1alpha, whereas BALB/c BMMCs significantly augmented their granule release. Lesions of BALB/c mice showed higher levels of degranulated mast cells at 3 h of infection, whereas after 3 days of infection, the number of degranulated mast cells in C57BL/6 was higher than in BALB/c lesions. Throughout infection, BALB/c mice harboured more parasites. The regulatory effect of mast cells seems to depend on the genetic background of the host: mast cells of BALB/c mice facilitate disease progression due to an augmented inflammatory response early in the infection, whereas mast cells of C57BL/6 mice produce cytokines that regulate inflammation and maintain an elevated number of immune cells in the lesions, promoting disease control. PMID- 18507783 TI - Towards a differential definition of atopy: Anisakis simplex and the relationship between parasites and arthropods in respiratory allergy. AB - Protective as well as enhancing effects of parasite infections on allergic disease have been postulated. Previous studies on this relationship focused frequently on skin test reactivity against aeroallergens, being house dust mites (HDM) the main agents responsible for a positive atopy outcome. We aimed to analyse the possible relationship between human parasite infection induced Anisakis simplex urticaria and respiratory allergy. A total of 86 patients with gastro-allergic Anisakiasis and 203 patients with chronic urticaria sensitized against A. simplex were studied for sensitization against aeroallergens and evaluated for rhinoconjunctivitis or bronchial asthma (RCBA). We compared the results with a control group of 250 consecutive patients referred for evaluation of allergic RCBA and atopy prevalence data of our region. Whereas no effect of A. simplex related disease on the overall allergic respiratory disease could be detected, a highly significant higher prevalence of RCBA associated HDM sensitization, but diminished allergy against other common aeroallergens (pollen, mould or dander) was observed in these groups. The relationship between A. simplex parasitism-associated acute or chronic urticaria on one side and allergic respiratory disease on the other side depends on the definition of atopy. We propose a differential definition of atopy, with a special emphasis on arthropod related sensitization. PMID- 18507784 TI - Toxocara canis larval excretory/secretory proteins impair eosinophil-dependent resistance of mice to Nippostrongylus brasiliensis. AB - Survival of parasitic helminths within a host requires immune evasion and excretory/secretory (ES) proteins may contribute to this process. Eosinophils are important effector cells in immunity of mice to the nematode Nippostrongylus brasiliensis and eosinophilic interleukin-5 transgenic (IL-5 Tg) mice are highly resistant to the earliest stages of primary infections. In contrast, Toxocara canis is largely resistant to eosinophils, with viable larvae encysted in tissues often surrounded by these and other leucocytes. The aim of this study was to investigate whether T. canis ES (TES) proteins inhibit eosinophil-dependent resistance to N. brasiliensis. Mouse serum pre-treated with TES had reduced capacity to mediate the adherence of leucocytes to N. brasiliensis infective stage larvae (L3) and this correlated with reduced complement C3 deposition on the parasite. TES did not inhibit eosinophil survival or eotaxin-dependent eosinophil migration in vitro. Cellular inflammation and eosinophil degranulation in the skin in response to injection of L3 was also not impaired by TES. However, when TES was included with L3 in an inoculum given to IL-5 Tg mice, a greatly increased number of parasites migrated to the lung. This suggests that the early eosinophil-dependent resistance in these mice was suppressed, by mechanisms yet to be determined. PMID- 18507785 TI - Association of early growth response-1 gene polymorphisms with total IgE and atopy in asthmatic children. AB - Early growth response-1 (Egr-1) is expressed in human airways and found to modulate tumor necrosis factor, immunoglobulin E (IgE), airway responsiveness, and interleukin-13-induced inflammation in mice. We investigated the effects of Chinese-tagging single nucleotide polymorphisms (SNPs) of Egr-1 on asthma traits in 298 Chinese asthmatic children and 175 controls, and a replication community cohort of 191 controls. Tag SNP (-4071 A-->G) and three additional SNPs (-1427 C- >T, -151 C-->T and IVS1 -42 C-->T) were genotyped by restriction fragment length polymorphism (RFLP). Significant associations were found between plasma total IgE concentration and -4071 A-->G (p = 0.008) and IVS1 -42 C-->T (p = 0.027) in asthmatic patients. After Bonferroni correction, only -4071 A-->G showed significant association. Multivariate regression analysis confirmed this significant association with a standardized coefficient beta of 0.156 (95% CI: 0.046-0.317; p = 0.009) in asthmatics among the three SNPs with age and gender adjusted. In -4071 A-->G, IgE(log) was significantly higher in patients with the GG genotype than the AA genotype (p = 0.009). In addition, -4071 A-->G was significantly associated with atopy (p = 0.016) and high total IgE concentration (p = 0.030) among asthmatics. Patients with the G allele had a 3.5-fold risk of having atopy and a 2.0-fold risk of having high total IgE concentration than those homozygous for the A allele. This is the first report to show significant association of Egr-1 polymorphisms with plasma total IgE and atopy in asthmatics. It may help to explore the pharmacogenetics of Egr-1 inhibitors. PMID- 18507786 TI - Quality of life, psychological adjustment and metabolic control in youths with type 1 diabetes: a study with self- and parent-report questionnaires. AB - OBJECTIVE: To evaluate self and parent reports on quality of life (QoL) and psychological adjustment of youths with type 1 diabetes, in comparison to a general paediatric population, and identify relationships between disease duration, metabolic control and psychological parameters. RESEARCH DESIGN AND METHODS: Participants included 70 youths with type 1 diabetes and their parents. They were compared with 70 non-diabetic subjects. Data were analyzed in the whole group and in subgroups aged 6-10, 11-13 and 14-18 yr. All cases performed pediatric QoL, Child Behaviour Checklist, filled in by parents, and Youth Self Report, filled in by youths. Data were compared with haemoglobin A1c (HbA1c) values and disease duration. RESULTS: Self-reports showed a psychological adjustment of youths with type 1 diabetes similar to that of controls. Parent reports showed that parents of children with type 1 diabetes were more worried than those of controls (p < 0.01). Adolescents showed a worse QoL and more frequent psychological disturbances. In this group, for youth and parent reports, HbA1c levels correlated positively with psychological problems (p < 0.05) and negatively with QoL (p < 0.05). Only for parent reports, in the whole group and in subgroups aged 6-10 and 11-13 yr, disease duration correlated positively with psychological adjustment (p < 0.05). CONCLUSIONS: Before adolescence, youths with type 1 diabetes showed only slight problems in psychological adjustment and QoL, with an association with disease duration reported by parents. In adolescence, both youths and their parents reported more emotional and behavioural problems, independent of disease duration. Better metabolic control and psychological well being seemed directly related. PMID- 18507787 TI - Maternal type 1 and gestational diabetes: postnatal differences in insulin secretion in offspring at preschool age. AB - BACKGROUND: It is well known that children born to mothers with diabetes in pregnancy are more likely to develop metabolic abnormalities in later life. Most prior studies have not differentiated between offspring of mothers with type 1 diabetes (T1DM) and gestational diabetes (GDM) or lack a control group of non exposed offspring. SUBJECTS: Offspring of T1DM (n = 16), GDM (n = 22) and mothers without diabetes (n = 25) born at Oulu University Hospital. AIM: To assess insulin secretion and insulin resistance in the offspring of T1DM and GDM at preschool age in comparison with offspring of non-diabetic mothers. METHODS: Anthropometric measurements and intravenous glucose tolerance testing were performed. First-phase insulin response (FPIR) and homoeostasis model assessment (HOMA) values were calculated. Pregnancy and birth data were analysed in relation to later metabolic parameters in all three groups using one-way analysis of variance (ANOVA) and analysis of covariance (ANCOVA). RESULTS: At a mean age of 4.9 yr, offspring of T1DM had increased fasting serum insulin concentrations (p = 0.044), FPIR (p = 0.034) and HOMA-B values (p = 0.008) compared with offspring of GDM or with offspring of healthy controls (statistically non-significant). The GDM gained least weight during pregnancy, and when adjusted for maternal weight gain during pregnancy, there were no statistically significant differences between study groups. CONCLUSIONS: Prenatal exposures to maternal type 1 and gestational diabetes may have different effects on postnatal glucose metabolism in the offspring assessed at a mean age close to 5 yr. Maternal weight gain in pregnancy may affect the postnatal glucose metabolism in the offspring. PMID- 18507788 TI - The triglyceride/HDL-cholesterol ratio as a marker of cardiovascular risk in obese children; association with traditional and emergent risk factors. AB - OBJECTIVES: To determine the presence of traditional and emergent cardiovascular risk factors and to evaluate the triglyceride/high-density lipoprotein cholesterol (Tg/HDL-C) ratio as a marker for cardiovascular disease and metabolic syndrome (MS) in obese children. MATERIAL AND METHODS: Sixty-seven prepubertal children of both sexes, between the ages of 6 and 12 yr, 20 normal-weight children, 18 overweight, and 29 obese subjects, were studied. Anthropometric measures, blood pressure, body mass index (BMI), and fat mass (FM), were measured. Plasma glucose, serum insulin, lipid profile, C-reactive protein (CRP), and leptin concentrations were quantified. Glucose and insulin concentrations 2 h post-glucose load were determined. The Tg/HDL-C ratio, homeostasis model assessment index (HOMA), and quantitative insulin sensitivity check index (QUICKI) were calculated. RESULTS: Systolic, diastolic, and mean blood pressures (MBP), low-density lipoprotein cholesterol (LDL-C), Tg/HDL-C, total cholesterol/HDL-C, LDL-C/HDL-C ratios, basal and 2 h postload insulin, CRP, and leptin were significantly higher and the QUICKI index were lower in the obese group. MBP, Tg/HDL-C ratio, HOMA, CRP, and leptin levels showed a positive and significant correlation and QUICKI a negative correlation with abdominal circumference, BMI, and FM. The Tg/HDL-C ratio correlated positively with MBP. The frequency of MS in the obese group was 69%. While Tg/HDL-C ratio, CRP, and leptin were higher and the values of QUICKI were lower in subjects with MS, it was the Tg/HDL-C ratio and the BMI that significantly explained the MS. CONCLUSIONS: Obesity increases the cardiovascular risk in childhood. The Tg/HDL-C ratio could be a useful index in identifying children at risk for dyslipidemia, hypertension, and MS. PMID- 18507789 TI - The HD-Zip IV gene GaHOX1 from cotton is a functional homologue of the Arabidopsis GLABRA2. AB - Most of the plant homeodomain-containing proteins play important roles in organ patterning and development, and Arabidopsis GLABRA2 (GL2), a member of the class IV homeodomain-leucine zipper (HD-ZIP) proteins, is a trichome and non-root hair cell regulator. Here we report the analysis of two cotton homeodomain-containing proteins, GaHOX1 and GaHOX2, isolated from the diploid cotton Gossypium arboreum. Both GaHOX1 and GaHOX2 belong to the class IV HD-ZIP family. When expressed under the control of the GL2 promoter, GaHOX1 rescued trichome development of an Arabidopsis glabrous mutant of gl2-2 (SALK_130213), whereas GaHOX2 did not. On the other hand, expression of GaHOX1 with a Cauliflower mosaic virus (CaMV) 35S promoter in the wild-type Arabidopsis plants suppressed the trichome development just as the GL2 ectopic expression. Expression analysis by Northern, RT-PCR and in situ hybridization indicated that GaHOX1 is predominantly expressed in cotton fiber cells at early developmental stages, consistent with its putative role in regulating cotton fiber development, while GaHOX2 is expressed in both fiber and other ovular tissues, including outer and inner integuments. Our results suggest that GaHOX1 is a functional homolog of GL2 in plant trichome development. PMID- 18507790 TI - How microspores transform into haploid embryos: changes associated with embryogenesis induction and microspore-derived embryogenesis. AB - Microspore embryogenesis is the most powerful androgenic pathway to produce haploid and doubled haploid plants. To deviate a microspore toward embryogenesis, a number of factors, different for each species, must concur at the same time and place. Once induced, the microspore undergoes numerous changes at different levels, from overall morphology to gene expression. Induction of microspore embryogenesis not only implies the expression of an embryogenic program, but also a stress-related cellular response and a repression of the gametophytic program to revert the microspore to a totipotent status. In this review, we compile the most recent advances in the understanding of the changes undergone by the induced microspore to readapt to the new developmental scenario. We devote special attention to the efforts made to uncover changes in the transcriptome of the induced microspore and microspore-derived embryo (MDE). Finally, we discuss the influence that an in vitro environment exerts over the MDE, as compared with its zygotic counterpart. PMID- 18507791 TI - The effect of cactus spines on light interception and Photosystem II for three sympatric species of Opuntia from the Mojave Desert. AB - Cactus spines reduce herbivory, direct water toward roots and reduce the impacts of high- and low-temperature extremes. Yet, shading of stems by spines reduces incident photosynthetic photon flux density (PFD), photosynthesis and growth. This study compared spinescence, PFD interception, stem temperature, Photosystem II (PSII) photochemistry and xanthophyll pigment composition for three species of cacti from the Mojave Desert, CA. The species vary in spinescence: Opuntia basilaris, which has no central or radial spines; Opuntia erinacea, which is densely covered with spines; and Opuntia phaeacantha, which has an intermediate coverage of spines. The role of spines was tested by removing spines from stems of O. erinacea. PFD interception was similar for both O. basilaris and O. phaeacantha, and about three times that for densely spined O. erinacea; removal of spines increased incident PFD three-fold. There were no effects of spines on stem temperatures. Steady-state light-response curves of chlorophyll a fluorescence from PSII indicated that PhiPSII, photochemical quenching (qP) and electron flux within PSII were lower, and non-photochemical quenching was higher, for O. erinacea in comparison to the other two species with less spines. After 2 months, qP was higher and electron flux lower, and xanthophyll pigment pool size was higher, for stems from which spines had been removed compared with intact stems. These three species allocate different amounts of biomass to spines, resulting in species-specific PFD interception, PSII photochemistry and xanthophyll pigment pool size, which may help maintain rates of photosynthesis during the hot, dry Mojave Desert summer. PMID- 18507792 TI - Towards molecular breeding of reproductive traits in cereal crops. AB - The transition from vegetative to reproductive phase, flowering per se, floral organ development, panicle structure and morphology, meiosis, pollination and fertilization, cytoplasmic male sterility (CMS) and fertility restoration, and grain development are the main reproductive traits. Unlocking their genetic insights will enable plant breeders to manipulate these traits in cereal germplasm enhancement. Multiple genes or quantitative trait loci (QTLs) affecting flowering (phase transition, photoperiod and vernalization, flowering per se), panicle morphology and grain development have been cloned, and gene expression research has provided new information about the nature of complex genetic networks involved in the expression of these traits. Molecular biology is also facilitating the identification of diverse CMS sources in hybrid breeding. Few Rf (fertility restorer) genes have been cloned in maize, rice and sorghum. DNA markers are now used to assess the genetic purity of hybrids and their parental lines, and to pyramid Rf or tms (thermosensitive male sterility) genes in rice. Transgene(s) can be used to create de novo CMS trait in cereals. The understanding of reproductive biology facilitated by functional genomics will allow a better manipulation of genes by crop breeders and their potential use across species through genetic transformation. PMID- 18507793 TI - Short or long day light regimes may not affect reproductive performance in the sow. AB - The objective of this study was to evaluate the effect of short or long day light regimes on the reproductive performance in the sow. The study comprised approximately 1300 sows and their 3400 breedings in three commercial sow pools, in which a batch farrowing of 40 sows was employed. Each batch remained in the farrowing unit for 8 weeks under either a short day (10 h light, 14 h dark) or a long day (14 h light, 10 h dark) light regime. After weaning and transportation to the central unit, all sows were kept under the long day regime until they were moved back to the farrowing unit. Production results for each individual sow were recorded. The effect of the two different light regimes on the farrowing rate (FR) and on the weaning-to-oestrous interval was evaluated using logistic and Poisson regression models, respectively. The light regime affected none of the parameters significantly. The FR in June, July and August did, however, drop below that in December, January and February (OR 0.7, p < 0.05). The FR was affected by the sow pool (p < 0.01). Parity two sows exhibited a lower FR (OR 0.5, p = 0.05) and higher incidence of delayed first oestrus (IRR 2.7, p < 0.01) than did older sows. This study indicates that despite an artificial light regime, sows may still react to changes in season. In conclusion, use of simple light regime to obtain complete control over reproductive performance appears to be difficult. PMID- 18507794 TI - Pregnancy-associated glycoprotein secretion in North Moroccan goats. AB - The plasma profile of pregnancy-associated glycoprotein (PAG) and its relationship with fetal number were studied in 14 North Moroccan goats using a semi-heterologous radioimmunoassay (RIA). Peripheral blood was collected every day during the first month of pregnancy, afterward the blood samples were collected trice a week. The PAG were first detected at day 20 of pregnancy, their levels increase to week 8 of gestation then decrease slightly until parturition. Statistical differences between goats carrying one or two foetuses are observed from week 5 of pregnancy until parturition. Factorial Discriminant Analysis provides mathematical models for the discrimination between the litter size using the PAG level and the week of pregnancy. Using only one blood sample per week, high sensitivity, specificity and overall accuracy (74%, 88% and 81%) were obtained using these models. It is concluded that the PAG RIA is an effective tool for early diagnosis of pregnancy and for discrimination between the litter sizes in North Moroccan goats. PMID- 18507795 TI - Evaluation of 'Section-Ligation-Release (SLR)' technique devised for castration in the stallion. AB - A novel technique [Section-Ligation-Release (SLR)] was evaluated for castration in the horse. Clinical traits, serum testosterone concentrations after challenge with human chorionic gonadotrophin (hCG), and histopathological changes of the testicular tissue were assessed. Five stallions, aged 24-48 months, were castrated using SLR technique under general anaesthesia. Both spermatic cords in each stallion were exposed at the scrotal neck by two 5-cm long incisions, followed by sharp dissection through the parietal vaginal tunic. Both vascular and non-vascular structures in the cords were triple clamped, transected and ligated. Both testes were left in situ. Serum testosterone concentrations were measured pre-operatively and at 2 months after castration following IV administration of 1 x 10(4) IU of hCG. Both testes from each castrate were collected at 2 months for histopathologic examination. SLR castration was successfully achieved. Moderated scrotal and preputial swelling was the only experienced short-term complication. Serum testosterone concentrations were significantly lower than basal pre-operative levels at 2 months after castration, and did not respond to hCG. On histopathology, hyalinization of the seminiferous tubules and loss of testicular interstitial tissue were indicative of complete avascular necrosis. This novel primary closure castration technique of stallion is a simple practical method, with minimal post-operative complications; and could be safely advocated as an alternative to the traditional castration techniques allowing for second intention healing of scrotal wounds. PMID- 18507796 TI - The relationship between linear type traits and fertility traits in high-yielding dairy goats. AB - Reproductive data collected from 13 medium-size goat farms in central Mexico (582 goats), were used to investigate if linear type traits influenced reproductive performance of high-yielding dairy goats. Data were analysed using multiple stepwise forward logistic regressions. Goats with the higher scores for strength were 1.43 times more likely (p < 0.05) to have larger litters (1.49 vs 1.39) than goats with lower strength. Goats approaching level rump angles were 1.68 times more likely (p < 0.01) to have larger litters, compared with goats with extremely sloped rump angles (1.48 vs 1.37). Goats with rear legs nearly straight (posty) in hock tended (p < 0.08) to presented larger litters than goats with rear legs slightly angled (sickled) in hock (1.49 vs 1.38). Less pronounced rump angle was favourable associated (p < 0.05) with litter weight (4.88 vs 4.53 kg) compared with goats with low pin bones. Goats with poorer udder texture were 1.42 more likely (p < 0.05) to have heavier litters than goats with good udder texture. Goats with faulty mammary system tended (p < 0.07) to have heavier litters than goats with good mammary system. Increasing udder height was associated (p < 0.05) with increasing number of services per conception. Goats with good fore udder attachment and non-refined heads were at reduced risk (p < 0.05) for stillbirths. Goats with stronger median ligament were 65% less likely (p < 0.05) to lose their foetuses than those with looser udder support. Our findings indicate that a greater litter size and litter weight is expected in goats as rump angle becomes less sloped. This study showed that a relationship exists between strength of goats and litter size, and that some traits linked to high milk yield were antagonist to some reproductive traits. PMID- 18507797 TI - Prostaglandin F 2 alpha stimulates endothelial nitric oxide synthase depending on the existence of bovine granulosa cells: analysis by co-culture system of endothelial cells, smooth muscle cells and granulosa cells. AB - Prostaglandin F(2 alpha) (PGF(2 alpha)) induces luteolysis in the mid but not in the early luteal phase; despite this, both the early and the mid corpus luteum (CL) have PGF(2 alpha) receptor (FPr). We previously indicated that the luteal blood flow surrounding the CL drastically increases prior to a decrease of progesterone (P) in the cows, suggesting that an acute increase of luteal blood flow may be an early sign of luteolysis in response to PGF(2 alpha) and that this may be induced by a vasorelaxant nitric oxide (NO). The aim of this study was to investigate the luteal stage-dependent and the site-restricted effect of PGF(2 alpha) and NO on the mRNA expressions and P secretion. To mimic the local luteal region both of peripheral and central areas of the CL, we utilized co-cultures using bovine aorta endothelial cells (EC), smooth muscle cells (SMC) and luteinizing granulosa cells (GC) or fully-luteinized GC. PGF(2 alpha) stimulated the expression of endothelial NO synthase (eNOS) mRNA at 0.5 h in mix-cultures of EC and SMC with fully-luteinized GC but not with luteinizing GC. The expression of eNOS mRNA in EC was increased by PGF(2 alpha) at 1 h only when EC was cultured together with fully-luteinized GC but not with luteinizing GC. In all co cultures, PGF(2 alpha) did not affect the mRNA expression of FPr. Treatment of NO donor inhibited P secretion at 0.5 h. In conclusion, the present study suggests that the coexistence of the mature luteal cells (fully-luteinized GC) with EC/SMC may be crucial for acquiring functional NO synthesis induced by PGF(2 alpha). PMID- 18507798 TI - A competitive dipstick enzyme immunoassay for diagnosis of early pregnancy in bovine. AB - Early pregnancy diagnosis in bovines is one of the important aspects in efficient dairy farm management. In order to develop a competitive enzyme immunoassay (EIA) employing low cost reagents, anti-progesterone antiserum and progesterone penicillinase enzyme conjugate were prepared. Using this anti-serum and conjugate along with pencillinV-starch-iodine substrate system, the competitive EIA was standardized. In the experiment, danazol, a weak androgen used to extract the progesterone bound to proteins in milk, was included after standardizing the optimum concentration. Incubation period and temperature and pH of the reaction mixture were also optimized. The developed test was validated with milk samples obtained from dairy farm and individual animal owners. Confirmation of the pregnancy was made by per rectum examination of the genital tract around 60 days post-insemination. The user friendly test procedure showed sensitivity and specificity of 83.3% and 87.5%, respectively as compared with residual binding method which was earlier developed in the laboratory with sensitivity and specificity of 100% and 87.5% respectively. PMID- 18507799 TI - Social dominance and courtship and mating behaviour in rams in non-competitive and competitive pen tests. AB - A group of 19 Merino rams was used to determine: (i) if body weight, age and testis characteristics are different between high (HR) and low-ranked (LR) rams, and (ii) how the competition between HR and LR rams for oestrous ewes affect their courtship and mating behaviour. Hierarchy was determined using the food competition test. The five first and five last ranked rams were individually exposed to oestrous ewes in pen tests, and the frequency of different courtship behavioural units, mounts and mates, and the time from test onset to each mate was recorded. The sexual behaviour of the five first ranked and the five last ranked rams was also tested in competition, pairing a HR and a LR ram. Age, body weight, body condition score, scrotal circumference, and testis firmness and resilience from HR and LR were not different. The display of courtship behavioural units was lower when rams were tested in competition but was not affected by the rank of the rams. The number of mates per test from HR rams was lower than that observed in LR rams when tested alone. While the number of mates decreased in LR rams in competition tests, it was not affected in HR rams. In summary, there was no relation between social rank and body weight, age or testis characteristics. LR rams were more effective than HR rams when mating oestrous ewes without competition. However, in competitive tests, although all rams modified their courtship strategies, LR rams were more affected by the presence of dominant rams than HR rams were by the presence of LR rams. PMID- 18507800 TI - Changes in testicular development, ultrasonographic and histological appearance of the testis in buck kids immunized against LHRH using recombinant LHRH fusion protein. AB - This study was designed to evaluate the effectiveness of recombinant Ovalbumin LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images. PMID- 18507801 TI - Sperm production and sperm morphology of Swedish Warmblood stallions. AB - The present study investigated daily sperm output and sperm morphology of fresh semen in eight Swedish Warmblood stallions aged 5-8 years. They were used for artificial insemination, and their fertility during the breeding season of semen collection exceeded 60% per cycle. One ejaculate of semen was collected daily for 10 consecutive days from each stallion. The gel-free volume was measured, and the sperm concentration was assessed with a Burker chamber. The volume of gel-free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). Sperm morphology was examined in ejaculates collected on days 2, 5 and 10. An aliquot from each ejaculate was fixed in 1 ml formol-saline immediately after collection and examined under a phase-contrast microscope (magnification 1000x) to assess morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin-eosin) for a more detailed examination of the sperm heads under a light microscope (magnification 1000x). Analysis of variance was applied to data. Total spermatozoa number decreased progressively during the first 8 days of collection, and daily sperm output (DSO) was calculated as mean TSN of collections on days 8-10, being 6.4 x 10(9) spermatozoa. The overall percentages of morphologically normal spermatozoa in ejaculates collected on days 2, 5 and 10 were above 70%, being significantly lower in ejaculate 2 (68.6%) compared with ejaculates 5 and 10 (72.9% respectively 75.3%). PMID- 18507802 TI - Effect of taurine and melatonin in the culture medium on buffalo in vitro embryo development. AB - This study was carried out to investigate the effect of supplementing culture medium with different concentrations of taurine and melatonin, on buffalo oocyte in vitro meiotic maturation and embryo development. In experiment 1, oocytes were matured in vitro and the cleaved embryos were cultured in the same following seven culture medium; (i) control (TCM 199 + 10% SS); (ii) control + 0.5 mM taurine; (iii) control + 1 mM taurine; (iv) control + 3 mM taurine; (v) control + 5 microM melatonin; (vi) control + 10 microM melatonin and (vii) control + 50 microM melatonin. In experiment 2, based on the results of experiment 1, to examine the synergistic effect of antioxidants, the oocytes were matured in culture medium (TCM199 + 10% SS), supplemented with both taurine at 1 mM and melatonin at 10 microM concentration and the cleaved embryos were cultured in the same medium. Supplementation of taurine at 1 mM concentration in the culture medium resulted in a higher (p < 0.05) transferable embryo (TE) yield when compared with control (20.6% vs 14.1%). Supplementation of melatonin at 10 and 50 microM concentration in the culture medium resulted in a higher (p < 0.05) meiotic maturation rate (90.3% and 88.8% respectively) and TE yield (28.4% and 27.2% respectively), than the other treatments. In experiment 2, the TE yield did not improve by supplementing the culture medium with both taurine and melatonin, when compared with melatonin alone. In conclusion, the results of this study demonstrated that, enriching the culture medium with taurine and melatonin, improves in vitro embryo production efficiency in buffaloes. In particular, a high TE yield was obtained by enriching the culture medium with 10 microM melatonin. PMID- 18507803 TI - The lack of alternative oxidase at low temperature leads to a disruption of the balance in carbon and nitrogen metabolism, and to an up-regulation of antioxidant defence systems in Arabidopsis thaliana leaves. AB - Alternative oxidase (AOX) catalyses the ATP-uncoupling cyanide (CN)-resistant pathway. In this study, our aim was to clarify the physiological role of AOX at low temperature. We examined the effect of low-temperature treatment on CN resistant respiration (CN-resistant R) and on the transcription of respiratory components in wild-type (WT) and aox1a knock-out transgenic (aox1a) Arabidopsis thaliana plants. In WT leaves, the expression of AOX1a mRNA was strongly induced by the low-temperature treatment, and thus CN-resistant R increased during low temperature treatment. In aox1a, the CN-sensitive respiration, and the expression of NDB2 and UCP1 were increased compared with WT. We compared several physiological parameters between WT and aox1a. Low-temperature treatment did not result in a visible phenotype to distinguish aox1a from WT. In aox1a, several antioxidant defence genes were induced, and the malondialdehyde content was lower than in WT. Starch content and a ratio of carbon to nitrogen were higher in aox1a than in WT. Our results indicate that a lack of AOX was linked to a difference in the carbon and nitrogen balance, and an up-regulation of the transcription of antioxidant defence system at low temperature. It is likely that AOX is a necessary component in antioxidant defence mechanisms and for the control of a balanced metabolism. PMID- 18507804 TI - Contractile roots in succulent monocots: convergence, divergence and adaptation to limited rainfall. AB - Contractile roots (CRs) that pull shoots further down in the soil are a possible example of convergent evolution in two monocot families, the Agavaceae and the Asphodelaceae. The association between CRs, water uptake and habitat aridity was investigated for agaves, yuccas and aloes by assessing the occurrence of CRs and the amount of root contraction for glasshouse-grown plants with respect to mean annual rainfall of their native habitats. Structural features of CRs as well as root hydraulic conductance were compared with those of non-contractile roots (NCRs). CRs occurred in 55% of the 73 species examined, including 64% of the agaves and 85% of the yuccas, but in none of the aloes despite the occurrence of CRs in related genera. The phylogenetic distribution of CRs was consistent with multiple acquisitions or losses of the trait. The amount of root contraction showed a highly significant negative relationship with mean annual rainfall, although other environmental factors may also be important. Radial hydraulic conductance of the basal (contractile) zone exceeded that of the midroot zone for CRs; for NCRs, the opposite was true. Thus, CRs in the species examined may provide a mechanism for greater water uptake near the soil surface in regions with limited rainfall. PMID- 18507805 TI - Abscisic acid signalling when soil moisture is heterogeneous: decreased photoperiod sap flow from drying roots limits abscisic acid export to the shoots. AB - To investigate the contribution of different parts of the root system to total sap flow and leaf xylem abscisic acid (ABA) concentration ([X-ABA](leaf)), individual sunflower (Helianthus annuus L.) shoots were grafted onto the root systems of two plants grown in separate pots and sap flow through each hypocotyl measured below the graft union. During deficit irrigation (DI), both pots received the same irrigation volumes, while during partial root zone drying (PRD) one pot ('wet') was watered and another ('dry') was not. During PRD, once soil water content (theta) decreased below a threshold, the fraction of sap flow from drying roots declined. As theta declined, root xylem ABA concentration increased in both irrigation treatments, and [X-ABA](leaf) increased in DI plants, but [X ABA](leaf) of PRD plants actually decreased within a certain theta range. A simple model that weighted ABA contributions of wet and dry root systems to [X ABA](leaf) according to the sap flow from each, better predicted [X-ABA](leaf) of PRD plants than either [X-ABA](dry), [X-ABA](wet) or their mean. Model simulations revealed that [X-ABA](leaf) during PRD exceeded that of DI with moderate soil drying, but continued soil drying (such that sap flow from roots in drying soil ceased) resulted in the opposite effect. PMID- 18507806 TI - Dynamic changes in the mitochondrial electron transport chain underpinning cold acclimation of leaf respiration. AB - We examined the effect of short- and long-term changes in temperature on gene expression, protein abundance, and the activity of the alternative oxidase and cytochrome oxidase pathways (AOP and COP, respectively) in Arabidopsis thaliana. The AOP was more sensitive to short-term changes in temperature than the COP, with partitioning to the AOP decreasing significantly below a threshold temperature of 20 degrees C. AOP activity was increased in leaves, which had been shifted to the cold for several days, but this response was transient, with AOP activity subsiding (and COP activity increasing) following the development of leaves in the cold. The transient increase in AOP activity in 10-d cold-shifted leaves was not associated with an increase in alternative oxidase (AOX) protein or AOX1a transcript abundance. By contrast, the amount of uncoupling protein was significantly increased in cold-developed leaves. In conjunction with this, transcript levels of the uncoupling protein-encoding gene UCP1 and the external NAD(P)H dehydrogenase-encoding gene NDB2 exhibited sustained increases following growth in the cold. The data suggest a role for each of these alternative non phosphorylating bypasses of mitochondrial electron transport at different points in time following exposure to cold, with increased AOP activity being important only in the early stages of cold treatment. PMID- 18507807 TI - Calcium pectate chemistry causes growth to be stored in Chara corallina: a test of the pectate cycle. AB - Calcium pectate chemistry was reported to control the growth rate of cells of Chara corallina, and required turgor pressure (P) to do so. Accordingly, this chemistry should account for other aspects of growth, particularly the ability of plants to compensate for brief exposure to low P, that is, to 'store' growth. Live Chara cells or isolated walls were attached to a pressure probe, and P was varied. Low P caused growth to be inhibited in live cells, but when P returned to normal (0.5 MPa), a flush of growth completely compensated for that lost at low P for as long as 23-53 min. This growth storage was absent in isolated walls, mature cells and live cells exposed to cold, indicating that the cytoplasm delivered a metabolically derived growth factor needing P for its action. Because the cytoplasm delivered pectate needing P for its action, pectate was supplied to isolated walls at low P as though the cytoplasm had done so. Growth was stored while otherwise none occurred. It was concluded that a P-dependent cycle of calcium pectate chemistry not only controlled growth rate and new wall deposition, but also accounted for stored growth. PMID- 18507808 TI - Jasmonic acid treatment to part of the root system is consistent with simulated leaf herbivory, diverting recently assimilated carbon towards untreated roots within an hour. AB - It is known that shoot application of jasmonic acid (JA) leads to an increased carbon export from leaves to stem and roots, and that root treatment with JA inhibits root growth. Using the radioisotope (11)C, we measured JA effects on carbon partitioning in sterile, split-root, barley plants. JA applied to one root half reduced carbon partitioning to the JA-treated tissue within minutes, whereas the untreated side showed a corresponding--but slower--increase. This response was not observed when instead of applying JA, the sink strength of one root half was reduced by cooling it: there was no enhanced partitioning to the untreated roots. The slower response in the JA-untreated roots, and the difference between the effect of JA and temperature, suggest that root JA treatment caused transduction of a signal from the treated roots to the shoot, leading to an increase in carbon allocation from the leaves to the untreated root tissue, as was indeed observed 10 min after the shoot application of JA. This supports the hypothesis that the response of some plant species to both leaf and root herbivores may be the diversion of resources to safer locations. PMID- 18507809 TI - Symbiosis between Nicotiana attenuata and Glomus intraradices: ethylene plays a role, jasmonic acid does not. AB - Phytohormones are thought to mediate plant-arbuscular mycorrhizal (AM) interactions. To explore the role of phytohormones in the interaction between Nicotiana attenuata and Glomus intraradices, we analysed levels of jasmonic acid (JA) and its amino acid conjugate JA-isoleucine/JA-leucine (JA-Ile), salicylic acid (SA) and ethylene in either infected or non-infected N. attenuata wild-type (WT) plants growing in soils that mimic the nutrient supply rates found in the plant's native environment. Under these conditions, the infection decreases plant growth and reproductive performance. Levels of JA, JA-Ile and SA did not change upon infection, but ethylene release was slightly decreased. Transgenic N. attenuata plants defective in JA signalling (aslox3 and ircoi1) did not differ significantly in growth or reproductive performance compared with infected WT. Furthermore, no difference in infection rates could be observed. Transgenic plants unable to produce (iraco) or perceive (etr1) ethylene showed significantly larger decreases in growth and number of seed capsules produced between infected and non-infected plants compared with WT plants. We conclude that ethylene, rather than JA, signalling plays a role in the interaction between N. attenuata and AM, from which the plant does not realize a fitness benefit. PMID- 18507810 TI - deltaO of water vapour, evapotranspiration and the sites of leaf water evaporation in a soybean canopy. AB - Stable isotopes in water have the potential to diagnose changes in the earth's hydrological budget in response to climate change and land use change. However, there have been few measurements in the vapour phase. Here, we present high frequency measurements of oxygen isotopic compositions of water vapour (delta(v)) and evapotranspiration (delta(ET)) above a soybean canopy using the tunable diode laser (TDL) technique for the entire 2006 growing season in Minnesota, USA. We observed a large variability in surface delta(v) from the daily to the seasonal timescales, largely explained by Rayleigh processes, but also influenced by vertical atmospheric mixing, local evapotranspiration (ET) and dew formation. We used delta(ET) measurements to calculate the isotopic composition at the sites of evaporative enrichment in leaves (delta(L,e)) and compared that with the commonly used steady-state prediction (delta(L,s)). There was generally a good agreement averaged over the season, but larger differences on individual days. We also found that vertical variability in relative humidity and temperature associated with canopy structure must be addressed in canopy-scale leaf water models. Finally, we explored this data set for direct evidence of the Peclet effect. PMID- 18507812 TI - Characterization of 19 novel cotton FLA genes and their expression profiling in fiber development and in response to phytohormones and salt stress. AB - Fasciclin-like arabinogalactan proteins (FLAs), a subclass of arabinogalactan proteins (AGPs), are usually involved in cell development in plants. To investigate the expression profiling as well as the role of FLA genes in fiber development, 19 GhFLA genes (cDNAs) were isolated from cotton (Gossypium hirsutum). Among them, 15 are predicted to be glycosylphosphatidylinositol anchored to the plasma membranes. The isolated cotton FLAs could be divided into four groups. Real-time quantitative reverse transcriptase polymerase chain reaction results indicated that the GhFLA genes are differentially expressed in cotton tissues. Three genes (GhFLA1/2/4) were specifically or predominantly expressed in 10 days post-anthesis fibers, and the transcripts of the other four genes (GhFLA6/14/15/18) were accumulated at relatively high levels in cotton fibers. Furthermore, expressions of the GhFLA genes are regulated in fiber development and in response to phytohormones and NaCl. The identification of cotton FLAs will facilitate the study of their roles in cotton fiber development and cell wall biogenesis. PMID- 18507811 TI - Trafficking of vesicular neurotransmitter transporters. AB - Vesicular neurotransmitter transporters are required for the storage of all classical and amino acid neurotransmitters in secretory vesicles. Transporter expression can influence neurotransmitter storage and release, and trafficking targets the transporters to different types of secretory vesicles. Vesicular transporters traffic to synaptic vesicles (SVs) as well as large dense core vesicles and are recycled to SVs at the nerve terminal. Some of the intrinsic signals for these trafficking events have been defined and include a dileucine motif present in multiple transporter subtypes, an acidic cluster in the neural isoform of the vesicular monoamine transporter (VMAT) 2 and a polyproline motif in the vesicular glutamate transporter (VGLUT) 1. The sorting of VMAT2 and the vesicular acetylcholine transporter to secretory vesicles is regulated by phosphorylation. In addition, VGLUT1 uses alternative endocytic pathways for recycling back to SVs following exocytosis. Regulation of these sorting events has the potential to influence synaptic transmission and behavior. PMID- 18507813 TI - Adjustments of water use efficiency by stomatal regulation during drought and recovery in the drought-adapted Vitis hybrid Richter-110 (V. berlandieri x V. rupestris). AB - The hybrid Richter-110 (Vitis berlandieri x Vitis rupestris) (R-110) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to water withholding followed by re-watering. The goal was to analyze how stomatal conductance (g(s)) is regulated with respect to different physiological variables under water stress and recovery, as well as how water stress affects adjustments of water use efficiency (WUE) at the leaf level. Water stress induced a substantial stomatal closure and an increase in WUE, which persisted many days after re-watering. The g(s) during water stress was mainly related to the content of ABA in the xylem and partly related to plant hydraulic conductivity but not to leaf water potential. By contrast, low g(s) during re watering did not correlate with ABA contents and was only related to a sustained decreased hydraulic conductivity. In addition to a complex physiological regulation of stomatal closure, g(s) and rate of transpiration (E) were strongly affected by leaf-to-air vapor pressure deficit (VPD) in a way dependent of the treatment. Interestingly, E increased with increasing VPD in control plants, but decreased with increasing VPD in severely stressed plants. All together, the fine stomatal regulation in R-110 resulted in very high WUE at the leaf level. This genotype is revealed to be very interesting for further studies on the physiological mechanisms leading to regulation of stomatal responsiveness and WUE in response to drought. PMID- 18507814 TI - Methane emission associated with anatomical and morphophysiological characteristics of rice (Oryza sativa) plant. AB - Plant-mediated transport is the primary route of methane (CH(4)) emission from the reduced paddy field to the aboveground atmosphere. Experiments were conducted at North Bank Plain Agro-climatic Zone of Assam, India, during monsoon rice growing season (July to December 2006) to elucidate the influences of anatomical and morphophysiological characteristics of rice (Oryza sativa L.) cultivars on methane emission from submerged agroecosystem. Ten rice cultivars were grown in light-textured loamy soil under rainfed uniform field condition. Among the 10 cultivars, 5 were traditional rice genotypes commonly grown in the agroclimatic zone and the other 5 were improved high-yielding varieties. Wide variation in CH(4) flux was recorded among the rice cultivars, which may be regulated by the difference in anatomical and morphophysiological characteristics of rice plant. Microscopic analysis of stem portion showed that high- and medium-CH(4)-emitting cultivars recorded higher size of the medullary cavity. Leaf area and transpirational rates were also found to be higher in high-CH(4)-emitting varieties. Scanning electron microscopic analysis revealed higher stomatal frequencies in high-methane-emitting cultivars. Data presented in this study suggest that variation in anatomical and morphophysiological characteristics among different rice genotypes may influence CH(4) emission from paddy fields. PMID- 18507815 TI - Impact of climate change on crop nutrient and water use efficiencies. AB - Implicit in discussions of plant nutrition and climate change is the assumption that we know what to do relative to nutrient management here and now but that these strategies might not apply in a changed climate. We review existing knowledge on interactive influences of atmospheric carbon dioxide concentration, temperature and soil moisture on plant growth, development and yield as well as on plant water use efficiency (WUE) and physiological and uptake efficiencies of soil-immobile nutrients. Elevated atmospheric CO(2) will increase leaf and canopy photosynthesis, especially in C3 plants, with minor changes in dark respiration. Additional CO(2) will increase biomass without marked alteration in dry matter partitioning, reduce transpiration of most plants and improve WUE. However, spatiotemporal variation in these attributes will impact agronomic performance and crop water use in a site-specific manner. Nutrient acquisition is closely associated with overall biomass and strongly influenced by root surface area. When climate change alters soil factors to restrict root growth, nutrient stress will occur. Plant size may also change but nutrient concentration will remain relatively unchanged; therefore, nutrient removal will scale with growth. Changes in regional nutrient requirements will be most remarkable where we alter cropping systems to accommodate shifts in ecozones or alter farming systems to capture new uses from existing systems. For regions and systems where we currently do an adequate job managing nutrients, we stand a good chance of continued optimization under a changed climate. If we can and should do better, climate change will not help us. PMID- 18507817 TI - Randomised controlled trial of a new palliative care service: Compliance, recruitment and completeness of follow-up. AB - BACKGROUND: Palliative care has been proposed for progressive non-cancer conditions but there have been few evaluations of service developments. We analysed recruitment, compliance and follow-up data of a fast track (or wait list control) randomised controlled trial of a new palliative care service - a design not previously used to assess palliative care. METHODS/DESIGN: An innovative palliative care service (comprising a consultant in palliative medicine, a clinical nurse specialist, an administrator and a psychosocial worker) was delivered to people severely affected by multiple sclerosis (MS), and their carers, in southeast London. Our design followed the MRC Framework for the Evaluation of Complex Interventions. In phase II we conducted randomised controlled trial, of immediate referral to the service (fast-track) versus a 12 week wait (standard best practice). Main outcome measures were: compliance (the extent the trial protocol was adhered to), recruitment (target 50 patients), attrition and missing data rates; trial outcomes were Palliative Care Outcome Scale and MS Impact Scale. RESULTS: 69 patients were referred, 52 entered the trial (26 randomised to each arm), 5 refused consent and 12 were excluded from the trial for other reasons, usually illness or urgent needs, achieving our target numbers. 25/26 fast track and 21/26 standard best practice patients completed the trial, resulting in 217/225 (96%) of possible interviews completed, 87% of which took place in the patient's home. Main reasons for failure to interview and/or attrition were death or illness. There were three deaths in the standard best practice group and one in the fast-track group during the trial. At baseline there were no differences between groups. Missing data for individual questionnaire items were small (median 0, mean 1-5 items out of 56+ items per interview), not associated with any patient or carer characteristics or with individual questionnaires, but were associated with interviewer. CONCLUSION: This is the first time a fast track (or wait list) randomised trial has been reported in palliative care. We found it achieved good recruitment and is a feasible method to evaluate palliative care services when patients are expected to live longer than 3-6 months. Home interviews are needed for a trial of this kind; interviewers need careful recruitment, training and supervision; and there should be careful separation from the clinical service of the control patients to prevent accidental contamination. TRIAL REGISTRATION: Clinical Trials.Gov NCT00364963. PMID- 18507818 TI - Long-term potentiation at C-fibre synapses by low-level presynaptic activity in vivo. AB - Inflammation, trauma or nerve injury trigger low-level activity in C-fibres and may cause long-lasting hyperalgesia. Long-term potentiation (LTP) at synapses of primary afferent C-fibres is considered to underlie some forms of hyperalgesia. In previous studies, high- but not low-frequency conditioning stimulation of C fibres has, however, been used to induce LTP in pain pathways. Recently we could show that also conditioning low-frequency stimulation (LFS) at C-fibre intensity induces LTP in vitro as well as in the intact animal, i.e. with tonic descending inhibition fully active. In the slice preparation, this form of LTP requires a rise in postsynaptic Ca2+-concentration and activation of Ca2+-dependent signalling pathways. Here, we investigated the signalling mechanisms underlying this novel form of LTP in vivo. We found that the signal transduction pathways causing LFS-induced LTP in vivo include activation of neurokinin 1 and N-methyl-D aspartate receptors, rise of [Ca2+]i from intracellular stores and via T-type voltage-dependent Ca2+ channels, activation of phospholipase C, protein kinase C and Ca2+-calmodulin dependent kinase II. These pathways match those leading to hyperalgesia in behaving animals and humans. We thus propose that LTP induced by low-level activity in C-fibres may underlie some forms of hyperalgesia. PMID- 18507819 TI - Do postage stamps versus pre-paid envelopes increase responses to patient mail surveys? A randomised controlled trial. AB - BACKGROUND: Studies largely from the market research field suggest that the inclusion of a stamped addressed envelope, rather than a pre-paid business reply, increases the response rate to mail surveys. The evidence that this is also the case regarding patient mail surveys is limited. METHODS: The aim of this study is to investigate whether stamped addressed envelopes increase response rates to patient mail surveys compared to pre-paid business reply envelopes and compare the relative costs. A sample of 477 initial non-responders to a mail survey of patients attending breast clinics in Greater Manchester between 1/10/2002 - 31/7/2003 were entered into the trial: 239 were randomly allocated to receive a stamped envelope and 238 to receive a pre-paid envelope in with their reminder surveys. Overall cost and per item returned were calculated. RESULTS: The response to the stamped envelope group was 31.8% (95% CI: 25.9% - 37.7%) compared to 26.9% (21.3% - 32.5%) for the pre-paid group. The difference (4.9% 95% CI: 3.3% - 13.1%) is not significant at alpha = 0.05 (chi2 = 1.39; 2 tailed test, d.f. = 1; P = 0.239). The stamped envelopes were cheaper in terms of cost per returned item (1.20 pounds) than the pre-paid envelopes (1.67 pounds). However if the set up cost for the licence to use the pre-paid service is excluded, the cost of the stamped envelopes is more expensive than pre-paid returns (1.20 pounds versus 0.73 pounds). CONCLUSION: Compared with pre-paid business replies, stamped envelopes did not produce a statistically significant increase in response rate to this patient survey. However, the response gain of the stamped strategy (4.9%) is similar to that demonstrated in a Cochrane review (5.3%) of strategies to increase response to general mail surveys. Further studies and meta analyses of patient responses to mail surveys via stamped versus pre-paid envelopes are needed with sufficient power to detect response gains of this magnitude in a patient population. PMID- 18507820 TI - The chromosomal passenger complex and the spindle assembly checkpoint: kinetochore-microtubule error correction and beyond. AB - During mitosis, correct bipolar chromosome attachment to the mitotic spindle is an essential prerequisite for the equal segregation of chromosomes. The spindle assembly checkpoint can prevent chromosome segregation as long as not all chromosome pairs have obtained bipolar attachment to the spindle. The chromosomal passenger complex plays a crucial role during chromosome alignment by correcting faulty chromosome-spindle interactions (e.g. attachments that do not generate tension). In the process of doing so, the chromosomal passenger complex generates unattached chromosomes, a specific situation that is known to promote checkpoint activity. However, several studies have implicated an additional, more direct role for the chromosomal passenger complex in enforcing the mitotic arrest imposed by the spindle assembly checkpoint. In this review, we discuss the different roles played by the chromosomal passenger complex in ensuring proper mitotic checkpoint function. Additionally, we discuss the possibility that besides monitoring the presence of unattached kinetochores, the spindle assembly checkpoint may also be capable of responding to chromosome-microtubule interactions that do not generate tension and we propose experimental set-ups to study this. PMID- 18507821 TI - Androgen receptor expresion in breast cancer: relationship with clinicopathological characteristics of the tumors, prognosis, and expression of metalloproteases and their inhibitors. AB - BACKGROUND: In the present study we analyze, in patients with breast cancer, the tumor expression of androgen receptors (AR), its relationship with clinicopathological characteristics and with the expression of several matrix metalloproteases (MMPs) and their inhibitors (TIMPs), as well as with prognosis. METHODS: An immunohistochemical study was performed using tissue microarrays and specific antibodies against AR, MMPs -1, -2, -7, -9, -11, -13, -14, and TIMPs -1, -2 and -3. More than 2,800 determinations on tumor specimens from 111 patients with primary invasive ductal carcinoma of the breast (52 with axillary lymph node metastases and 59 without them) and controls were performed. Staining results were categorized using a score based on the intensity of the staining and a specific software program calculated the percentage of immunostained cells automatically. RESULTS: A total of 83 cases (74.8%) showed a positive immunostaining for AR, but with a wide variation in the staining score values. There were no significant associations between the total immunostaining scores for AR and any clinicopathological parameters. However, score values for MMP-1, 7 and -13, were significantly higher in AR-positive tumors than in AR-negative tumors. Likewise, when we considered the cellular type expressing each factor, we found that AR-positive tumors had a higher percentage of cases positive for MMP 1, -7, -11, and TIMP-2 in their malignant cells, as well as for MMP-1 in intratumoral fibroblasts. On the other hand, multivariate analysis demonstrated that patients with AR-positive tumors have a significant longer overall survival than those with AR-negative breast carcinomas (p = 0.03). CONCLUSION: Our results confirm that AR are commonly expressed in breast cancer, and are correlated with the expression of some MMPs and TIMP-2. Although we found a specific value of AR expression to be a prognostic indicator in breast cancer, the functional role of AR in these neoplasms is still unclear and further data are needed in order to clarify their biological signification in breast cancer. PMID- 18507822 TI - Unraveling the genomic mosaic of a ubiquitous genus of marine cyanobacteria. AB - BACKGROUND: The picocyanobacterial genus Synechococcus occurs over wide oceanic expanses, having colonized most available niches in the photic zone. Large scale distribution patterns of the different Synechococcus clades (based on 16S rRNA gene markers) suggest the occurrence of two major lifestyles ('opportunists'/'specialists'), corresponding to two distinct broad habitats ('coastal'/'open ocean'). Yet, the genetic basis of niche partitioning is still poorly understood in this ecologically important group. RESULTS: Here, we compare the genomes of 11 marine Synechococcus isolates, representing 10 distinct lineages. Phylogenies inferred from the core genome allowed us to refine the taxonomic relationships between clades by revealing a clear dichotomy within the main subcluster, reminiscent of the two aforementioned lifestyles. Genome size is strongly correlated with the cumulative lengths of hypervariable regions (or 'islands'). One of these, encompassing most genes encoding the light-harvesting phycobilisome rod complexes, is involved in adaptation to changes in light quality and has clearly been transferred between members of different Synechococcus lineages. Furthermore, we observed that two strains (RS9917 and WH5701) that have similar pigmentation and physiology have an unusually high number of genes in common, given their phylogenetic distance. CONCLUSION: We propose that while members of a given marine Synechococcus lineage may have the same broad geographical distribution, local niche occupancy is facilitated by lateral gene transfers, a process in which genomic islands play a key role as a repository for transferred genes. Our work also highlights the need for developing picocyanobacterial systematics based on genome-derived parameters combined with ecological and physiological data. PMID- 18507823 TI - Patients with rheumatoid arthritis have an altered circulatory aggrecan profile. AB - BACKGROUND: Rheumatoid arthritis (RA) is a chronic auto-immune disease with extensive articular cartilage destruction. Aggrecan depletion, mediated by aggrecanases is one of the first signs of early cartilage erosion. We investigated, whether measurement of aggrecan and fragments thereof in serum, could be used as biomarkers for joint-disease in RA patients and furthermore characterized the fragments found in the circulation. METHODS: The study consisted of 38 patients, 12 males (62.2 +/- 16.0 years) and 26 females (59.8 +/- 20.7 years) diagnosed with RA: 41.5 +/- 27.5 mm/h erythrocyte sedimentation rate (ESR), 38.4 +/- 34.7 mg/ml C-reactive protein (CRP) and 4.8 +/- 1.7 disease activity score (DAS) and 108 healthy age-matched controls. Aggrecan levels were measured using two immunoassays, i.e. the (374)ARGSVI-G2 sandwich ELISA measuring aggrecanase-mediated aggrecan degradation and the G1/G2 sandwich assay, detecting aggrecan molecules containing G1 and/or G2 (total aggrecan) We further characterized serum samples by western blots, by using monoclonal antibodies F 78, binding to G1 and G2, or by BC-3, detecting the aggrecanase-generated N terminal 374ARGSVI neo-epitope. RESULTS: Total aggrecan levels in RA patients were significantly decreased from 824.8 +/- 31 ng/ml in healthy controls to 570.5 +/- 30 ng/ml (31% decrease, P < 0.0001), as measured by the G1/G2 ELISA. Western blot analysis with F-78 showed one strong band at 10 kDa, and weaker bands at 25 and 45 kDa in both healthy controls and RA patients. In contrast, staining for aggrecanase-activity revealed only one strong band in RA patients of 45 kDa. CONCLUSION: This is the first study, which characterizes different aggrecan fragments in human serum. The data strongly suggests that total aggrecan levels, i.e. aggrecan molecules containing G1 and/or G2 are lower in RA patients, and that RA patients have at least one specific subpopulation of aggrecan fragments, namely aggrecanse generated 374ARGSVI fragments. Further clinical studies are needed to investigate the potential of G1/G2 as a structure-related biochemical marker in destructive joint-diseases. PMID- 18507824 TI - Control over the morphology and segregation of Zebrafish germ cell granules during embryonic development. AB - BACKGROUND: Zebrafish germ cells contain granular-like structures, organized around the cell nucleus. These structures share common features with polar granules in Drosophila, germinal granules in Xenopus and chromatoid bodies in mice germ cells, such as the localization of the zebrafish Vasa, Piwi and Nanos proteins, among others. Little is known about the structure of these granules as well as their segregation in mitosis during early germ-cell development. RESULTS: Using transgenic fish expressing a fluorescently labeled novel component of Zebrafish germ cell granules termed Granulito, we followed the morphology and distribution of the granules. We show that whereas these granules initially exhibit a wide size variation, by the end of the first day of development they become a homogeneous population of medium size granules. We investigated this resizing event and demonstrated the role of microtubules and the minus-end microtubule dependent motor protein Dynein in the process. Last, we show that the function of the germ cell granule resident protein the Tudor domain containing protein-7 (Tdrd7) is required for determination of granule morphology and number. CONCLUSION: Our results suggest that Zebrafish germ cell granules undergo a transformation process, which involves germ cell specific proteins as well as the microtubular network. PMID- 18507825 TI - Interactions of biotic and abiotic environmental factors in an ectomycorrhizal symbiosis, and the potential for selection mosaics. AB - BACKGROUND: Geographic selection mosaics, in which species exert different evolutionary impacts on each other in different environments, may drive diversification in coevolving species. We studied the potential for geographic selection mosaics in plant-mycorrhizal interactions by testing whether the interaction between bishop pine (Pinus muricata D. Don) and one of its common ectomycorrhizal fungi (Rhizopogon occidentalis Zeller and Dodge) varies in outcome, when different combinations of plant and fungal genotypes are tested under a range of different abiotic and biotic conditions. RESULTS: We used a 2 x 2 x 2 x 2 factorial experiment to test the main and interactive effects of plant lineage (two maternal seed families), fungal lineage (two spore collections), soil type (lab mix or field soil), and non-mycorrhizal microbes (with or without) on the performance of plants and fungi. Ecological outcomes, as assessed by plant and fungal performance, varied widely across experimental environments, including interactions between plant or fungal lineages and soil environmental factors. CONCLUSION: These results show the potential for selection mosaics in plant mycorrhizal interactions, and indicate that these interactions are likely to coevolve in different ways in different environments, even when initially the genotypes of the interacting species are the same across all environments. Hence, selection mosaics may be equally as effective as genetic differences among populations in driving divergent coevolution among populations of interacting species. PMID- 18507826 TI - Organelle-specific isoenzymes of plant V-ATPase as revealed by in vivo-FRET analysis. AB - BACKGROUND: The V-ATPase (VHA) is a protein complex of 13 different VHA-subunits. It functions as an ATP driven rotary-motor that electrogenically translocates H+ into endomembrane compartments. In Arabidopsis thaliana V-ATPase is encoded by 23 genes posing the question of specific versus redundant function of multigene encoded isoforms. RESULTS: The transmembrane topology and stoichiometry of the proteolipid VHA-c" as well as the stoichiometry of the membrane integral subunit VHA-e within the V-ATPase complex were investigated by in vivo fluorescence resonance energy transfer (FRET). VHA-c", VHA-e1 and VHA-e2, VHA-a, VHA-c3, truncated variants of VHA-c3 and a chimeric VHA-c/VHA-c" hybrid were fused to cyan (CFP) and yellow fluorescent protein (YFP), respectively. The constructs were employed for transfection experiments with Arabidopsis thaliana mesophyll protoplasts. Subcellular localization and FRET analysis by confocal laser scanning microscopy (CLSM) demonstrated that (i.) the N- and C-termini of VHA-c" are localised in the vacuolar lumen, (ii.) one copy of VHA-c" is present within the VHA-complex, and (iii.) VHA-c" is localised at the ER and associated Golgi bodies. (iv.) A similar localisation was observed for VHA-e2, whereas (v.) the subcellular localisation of VHA-e1 indicated the trans Golgi network (TGN) specifity of this subunit. CONCLUSION: The plant proteolipid ring is a highly flexible protein subcomplex, tolerating the incorporation of truncated and hybrid proteolipid subunits, respectively. Whereas the membrane integral subunit VHA-e is present in two copies within the complex, the proteolipid subunit VHA-c" takes part in complex formation with only one copy. However, neither VHA-c" isoform 1 nor any of the two VHA-e isoforms were identified at the tonoplast. This suggest a function in endomembrane specific VHA-assembly or targeting rather than proton transport. PMID- 18507827 TI - Overexpression of Pax6 results in microphthalmia, retinal dysplasia and defective retinal ganglion cell axon guidance. AB - BACKGROUND: The transcription factor Pax6 is expressed by many cell types in the developing eye. Eyes do not form in homozygous loss-of-function mouse mutants (Pax6Sey/Sey) and are abnormally small in Pax6Sey/+ mutants. Eyes are also abnormally small in PAX77 mice expressing multiple copies of human PAX6 in addition to endogenous Pax6; protein sequences are identical in the two species. The developmental events that lead to microphthalmia in PAX77 mice are not well characterised, so it is not clear whether over- and under-expression of Pax6/PAX6 cause microphthalmia through similar mechanisms. Here, we examined the consequences of over-expression for the eye and its axonal connections. RESULTS: Eyes form in PAX77+/+ embryos but subsequently degenerate. At E12.5, we found no abnormalities in ocular morphology, retinal cell cycle parameters and the incidence of retinal cell death. From E14.5 on, we observed malformations of the optic disc. From E16.5 into postnatal life there is progressively more severe retinal dysplasia and microphthalmia. Analyses of patterns of gene expression indicated that PAX77+/+ retinae produce a normal range of cell types, including retinal ganglion cells (RGCs). At E14.5 and E16.5, quantitative RT-PCR with probes for a range of molecules associated with retinal development showed only one significant change: a slight reduction in levels of mRNA encoding the secreted morphogen Shh at E16.5. At E16.5, tract-tracing with carbocyanine dyes in PAX77+/+ embryos revealed errors in intraretinal navigation by RGC axons, a decrease in the number of RGC axons reaching the thalamus and an increase in the proportion of ipsilateral projections among those RGC axons that do reach the thalamus. A survey of embryos with different Pax6/PAX6 gene dosage (Pax6Sey/+, Pax6+/+, PAX77+ and PAX77+/+) showed that (1) the total number of RGC axons projected by the retina and (2) the proportions that are sorted into the ipsilateral and contralateral optic tracts at the optic chiasm vary differently with gene dosage. Increasing dosage increases the proportion projecting ipsilaterally regardless of the size of the total projection. CONCLUSION: Pax6 overexpression does not obviously impair the initial formation of the eye and its major cell-types but prevents normal development of the retina from about E14.5, leading eventually to severe retinal degeneration in postnatal life. This sequence is different to that underlying microphthalmia in Pax6+/- heterozygotes, which is due primarily to defects in the initial stages of lens formation. Before the onset of severe retinal dysplasia, Pax6 overexpression causes defects of retinal axons, preventing their normal growth and navigation through the optic chiasm. PMID- 18507828 TI - Development and preliminary evaluation of the participation in life activities scale for children and adolescents with asthma: an instrument development study. AB - BACKGROUND: Being able to do things other kids do is the desire of school-age children and adolescents with asthma. In a phenomenology study, adolescents identified participation in life activities as the outcome variable and primary motivator for behavioral changes in coming to accept asthma as a chronic condition. In preparation for testing an acceptance model for older school-age children and early adolescents diagnosed with asthma, the Participation in Life Activities Scale was developed. The purposes of this paper are to describe development, and report on face and content validity of the scale designed to measure one aspect of quality of life defined as level of unrestricted involvement in chosen pursuits. METHODS: Items generated for the instrument evolved from statements and themes extracted from qualitative interviews. Face and content validity were evaluated by eight lay reviewers and 10 expert reviewers. Rate of accurate completion was computed using a convenience, cross section sample consisting of 313 children and adolescents with asthma, ages 9-15 years, drawn from three studies. Preliminary cross-group comparisons of scores were assessed using t-tests and analysis of variance. RESULTS: Face and content validity were determined to be highly acceptable and relevant, respectively. Completion rate across all three studies was 97%. Although cross-group comparisons revealed no significant differences in overall participation scores based on age, race or residence groupings (p > .05), significant difference were indicated between males and females (p = .02), as well as the highest and lowest socioeconomic groups (p = .002). CONCLUSION: Assessing content validity was the first step in evaluating properties of this newly developed instrument. Once face and content validity were established, psychometric evaluation related to internal consistency reliability and construct validity using factor analysis procedures was begun. Results will be reported elsewhere. PMID- 18507829 TI - Optimization in computational systems biology. AB - Optimization aims to make a system or design as effective or functional as possible. Mathematical optimization methods are widely used in engineering, economics and science. This commentary is focused on applications of mathematical optimization in computational systems biology. Examples are given where optimization methods are used for topics ranging from model building and optimal experimental design to metabolic engineering and synthetic biology. Finally, several perspectives for future research are outlined. PMID- 18507830 TI - Could a defective epithelial sodium channel lead to bronchiectasis. AB - BACKGROUND: Bronchiectasis is defined as a permanent dilation of the airways arising from chronic bronchial inflammation/infection. In 50% of cases, no etiology can be identified. Recently, the role of the epithelial sodium channel ENaC has been pointed out in the pathophysiology of cystic fibrosis, a disease due to mutations in the CFTR gene and causing bronchiectasis in the airways. Moreover, it was found that transgenic mice overexpressing ENaCbeta present cystic fibrosis-like lung disease symptoms. Our aim was to evaluate if a defective ENaC protein could be involved in the development of bronchiectasis. METHODS: We extensively analysed ENaCbeta and gamma genes in 55 patients with idiopathic bronchiectasis and without two mutations in the coding regions of CFTR. Thirty-eight patients presented functional abnormalities suggesting impaired sodium transport (abnormal sweat chloride concentration or nasal potential difference measurement), and 17 had no such evidence. RESULTS: Sequencing of the exons and flanking introns of the ENaCbeta and gamma gene identified five different amino-acid changes (p.Ser82Cys, p.Pro369Thr, p.Asn288Ser in ENaCbeta ; and p.Gly183Ser, p.Glu197Lys in ENaCgamma) in heterozygous state in 8 patients. The p.Ser82Cys amino-acid change was found in 3 unrelated patients who were also heterozygous for a CFTR mutation or variant (1 p.F508del, 1 IVS8-5T, and 1 IVS8-5T:1716G>A (p.E528E)). The other mutations were found in patients without CFTR mutation, the p.Glu197Lys mutation in 2 patients and the other variants in single patients. Among the 8 patients bearing an ENaC mutation, 5 had functional abnormalities suggesting impaired sodium transport. CONCLUSION: Our results suggest that several variants in ENaCbeta and gamma genes might be deleterious for ENaC function and lead to bronchiectasis, especially in patients who are trans-heterozygotes for ENaCbeta/CFTR mutations or variants. PMID- 18507831 TI - Senescence marker protein 30 in acute liver failure: validation of a mass spectrometry proteomics assay. AB - BACKGROUND: Our previous proteomic study showed that the senescence marker protein (SMP30) is selectively present in the plasma of a murine model of acute liver failure (ALF). The aim of this study was to validate this SMP30 expression in the plasma and liver tissues of mice and humans with ALF. METHODS: After the proteomic analysis of plasma from a murine model of D galactosamine/lipopolysaccharide (GalN/LPS)-induced ALF by two-dimensional electrophoresis (2-DE) and mass spectrometry, the expression levels of SMP30 in the plasma and liver tissues were validated by western blot and RT-PCR analyses. These results were then confirmed in plasma samples from humans. RESULTS: These data validate the results of 2-DE, and western blot showed that SMP30 protein levels were only elevated in the plasma of ALF mice. Further analysis revealed that GalN/LPS induced the downregulation of SMP30 protein levels in liver tissues (by approximately 25% and 16% in the GalN/LPS-treated mice and in the treated mice that survived, respectively; P < 0.01). Hepatic SMP30 mRNA levels decreased by about 90% only in the mice that survived the GalN/LPS treatment. Importantly, plasma obtained from patients with ALF also contained higher levels of SMP30, about (3.65 +/- 0.34) times those observed in healthy volunteers. CONCLUSION: This study shows that SMP30 is not only a potential biomarker for the diagnosis and even prognosis of ALF. It also plays a very important role in a self protective mechanism in survival and participates in the pathophysiological processes of ALF. PMID- 18507832 TI - Prevalence of A2143G mutation of H. pylori-23S rRNA in Chinese subjects with and without clarithromycin use history. AB - BACKGROUND: A2143G mutation of 23S rRNA gene of H. pylori results in clarithromycin (CLR) resistance. To investigate the prevalence of the CLR resistance-related A2143G mutation of the H. pylori-specific 23S rRNA gene in Chinese subjects with and without CLR use history, 307 subjects received the treatment with amoxicillin and omeprazole (OA) and 310 subjects received a placebo in 1995, and 153 subjects received a triple therapy with OA and CLR (OAC) in 2000. DNA was extracted from fasting gastric juice at the end of the intervention trial in 2003. H. pylori infection was determined by H. pylori specific 23S rRNA PCR, ELISA, and13C-urea breath test assays. Mutations of the 23S rRNA gene were detected by RFLP assays. RESULTS: The presence of 23S rRNA due to H. pylori infection in the OA group remained lower than that in the placebo group 7.3 yrs after OA-therapy [51.1% (157/307) vs. 83.9% (260/310), p = 0.0000]. In the OAC group, the 23S rRNA detection rate was 26.8% (41/153) three yrs after OAC-treatment. The A2143G mutation rate among the 23S rRNA-positive subjects in the OAC group [31.7% (13/41)] was significantly higher than that in the OA group [10.2% (16/157)] and the placebo group [13.8% (36/260)]. The frequency of the AAGGG --> CTTCA (2222-2226) and AACC --> GAAG (2081-2084) sequence alterations in the OAC group was also significantly higher than those in the OA group and the placebo group. CONCLUSION: Primary prevalence of the A2143G mutation was 10~14% among Chinese population without history of CLR therapy. Administration of CLR to eliminate H. pylori infection increased the prevalence of the A2143G mutation in Chinese subjects (32%) significantly. PMID- 18507833 TI - Use of residential wood heating in a context of climate change: a population survey in Quebec (Canada). AB - BACKGROUND: Wood heating is recommended in several countries as a climate change (CC) adaptation measure, mainly to increase the autonomy of households during power outages due to extreme climatic events. The aim of this study was to examine various perceptions and individual characteristics associated with wood heating through a survey about CC adaptations. METHODS: A telephone survey (n = 2,545) of adults living in the southern part of the province of Quebec (Canada) was conducted in the early fall season of 2005. The questionnaire used closed questions and measured the respondents' beliefs and current adaptations about CC. Calibration weighting was used to adjust the data analysis for the respondent's age and language under stratified sampling based on health regions. RESULTS: More than three out of four respondents had access to a single source of energy at home, which was mainly electricity; 22.2% combined two sources or more; 18.5% heated with wood occasionally or daily during the winter. The prevalence of wood heating was higher in the peripheral regions than in the more urban regions, where there was a higher proportion of respondents living in apartments. The prevalence was also higher with participants completely disagreeing (38.5%) with the eventual prohibition of wood heating when there is smog in winter, compared to respondents somewhat disagreeing (24.2%) or agreeing (somewhat: 17.5%; completely: 10.4%) with the adoption of this strategy. It appears that the perception of living in a region susceptible to winter smog, smog warnings in the media, or the belief in the human contribution to CC, did not influence significantly wood heating practices. CONCLUSION: Increased residential wood heating could very well become a maladaptation to climate change, given its known consequences on winter smog and respiratory health. It would thus be appropriate to implement a long-term national program on improved and controlled residential wood heating. This would constitute a "no-regrets" adaptation to climate change, while reducing air pollution and its associated health impacts. PMID- 18507834 TI - Enhancement of in vitro interleukin-2 production in normal subjects following a single spinal manipulative treatment. AB - BACKGROUND: Increasing evidence supports somato-visceral effects of manual therapies. We have previously demonstrated that a single spinal manipulative treatment (SMT) accompanied by audible release has an inhibitory effect on the production of proinflammatory cytokines in asymptomatic subjects. The purpose of this study is to report on SMT-related changes in the production of the immunoregulatory cytokine interleukin 2 (IL-2) and to investigate whether such changes might differ with respect to the treatment approach related to the presence or absence of an audible release (joint cavitation). METHODS: Of 76 asymptomatic subjects, 29 received SMT with cavitation (SMT-C), 23 were treated with SMT without cavitation (SMT-NC) and 24 comprised the venipuncture control (VC) group. The SMT-C and SMT-NC subjects received a single, similar force high velocity low amplitude manipulation, in the upper thoracic spine. However, in SMT NC subjects, positioning and line of drive were not conducive to cavitation. Blood and serum samples were obtained before and then at 20 and 120 min post intervention. The production of IL-2 in peripheral blood mononuclear cell cultures was induced by activation for 48 hr with Staphylococcal protein A (SPA) and, in parallel preparations, with the combination of phorbol ester (TPA) and calcium ionophore. The levels of IL-2 in culture supernatants and serum were assessed by specific immunoassays. RESULTS: Compared with VC and their respective baselines, SPA-induced secretion of IL-2 increased significantly in cultures established from both SMT-C and SMT-NC subjects at 20 min post-intervention. At 2 hr post-treatment, significant elevation of IL-2 synthesis was still apparent in preparations from SMT-treated groups though it became somewhat attenuated in SMT NC subjects. Conversely, IL-2 synthesis induced by TPA and calcium ionophore was unaltered by either type of SMT and was comparable to that in VC group at all time points. No significant alterations in serum-associated IL-2 levels were observed in any of the study groups. CONCLUSION: The present study demonstrates that, the in vitro T lymphocyte response to a conventional mitogen (SPA), as measured by IL-2 synthesis, can become enhanced following SMT. Furthermore, within a period of time following the manipulative intervention, this effect may be independent of joint cavitation. Thus the results of this study suggest that, under certain physiological conditions, SMT might influence IL-2-regulated biological responses. PMID- 18507835 TI - Assessment of decorin-binding protein A to the infectivity of Borrelia burgdorferi in the murine models of needle and tick infection. AB - BACKGROUND: Decorin-binding proteins (Dbps) A and B of Borrelia burgdorferi, the agent of Lyme disease, are surface-exposed lipoproteins that presumably bind to the extracellular matrix proteoglycan, decorin. B. burgdorferi infects various tissues including the bladder, heart, joints, skin and the central nervous system, and the ability of B. burgdorferi to bind decorin has been hypothesized to be important for this disseminatory pathogenic strategy. RESULTS: To determine the role of DbpBA in the infectious lifecycle of B. burgdorferi, we created a DbpBA-deficient mutant of B. burgdorferi strain 297 and compared the infectious phenotype of the mutant to the wild-type strain in the experimental murine model of Lyme borreliosis. The mutant strain exhibited a 4-log decrease in infectivity, relative to the wild-type strain, when needle inoculated into mice. Upon complementation of the DbpBA-mutant strain with DbpA, the wild-type level of infectivity was restored. In addition, we demonstrated that the DbpBA-deficient mutant was able to colonize Ixodes scapularis larval ticks after feeding on infected mice and persist within the ticks during the molt to the nymphal state. Moreover, surprisingly, the DbpBA-mutant strain was capable of being transmitted to naive mice via tick bite, giving rise to infected mice. CONCLUSION: These results suggest that DbpBA is not required for the natural tick-transmission process to mammals, despite inferences from needle-inoculation experiments implying a requirement for DbpBA during mammalian infection. The combined findings also send a cautionary note regarding how results from needle inoculation experiments with mice should be interpreted. PMID- 18507836 TI - Identification and characteristics of microRNAs from Bombyx mori. AB - BACKGROUND: MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression by targeting messenger RNAs (mRNAs) and causing mRNA cleavage or translation blockage. Of the 355 Arthropod miRNAs that have been identified, only 21 are B. mori miRNAs that were predicted computationally; of these, only let-7 has been confirmed by Northern blotting. RESULTS: Combining a computational method based on sequence homology searches with experimental identification based on microarray assays and Northern blotting, we identified 46 miRNAs, an additional 21 plausible miRNAs, and a novel small RNA in B. mori. The latter, bmo miR-100-like, was identified using the known miRNA aga-miR-100 as a probe; bmo miR-100-like was detected by microarray assay and Northern blotting, but its precursor sequences did not fold into a hairpin structure. Among these identified miRNAs, we found 12 pairs of miRNAs and miRNA*s. Northern blotting revealed that some B. mori miRNA genes were expressed only during specific stages, indicating that B. mori miRNA genes (e.g., bmo-miR-277) have developmentally regulated patterns of expression. We identified two miRNA gene clusters in the B. mori genome. bmo-miR-2b, which is found in the gene cluster bmo-miR-2a-1/bmo-miR-2a 1*/bmo-miR-2a-2/bmo-miR-2b/bmo-miR-13a*/bmo-miR-13b, encodes a newly identified member of the mir-2 family. Moreover, we found that methylation can increase the sensitivity of a DNA probe used to detect a miRNA by Northern blotting. Functional analysis revealed that 11 miRNAs may regulate 13 B. mori orthologs of the 25 known Drosophila miRNA-targeted genes according to the functional conservation. We predicted the binding sites on the 1671 3'UTR of B. mori genes; 547 targeted genes, including 986 target sites, were predicted. Of these target sites, 338 had perfect base pairing to the seed region of 43 miRNAs. From the predicted genes, 61 genes, each of them with multiple predicted target sites, should be considered excellent candidates for future functional studies. Biological classification of predicted miRNA targets showed that "binding", "catalytic activity" and "physiological process" were over-represented for the predicted genes. CONCLUSION: Combining computational predictions with microarray assays, we identified 46 B. mori miRNAs, 13 of which were miRNA*s. We identified a novel small RNA and 21 plausible B. mori miRNAs that could not be located in the available B. mori genome, but which could be detected by microarray. Thirteen and 547 target genes were predicted according to the functional conservation and binding sites, respectively. Identification of miRNAs in B. mori, particularly those that are developmentally regulated, provides a foundation for subsequent functional studies. PMID- 18507837 TI - Effects of common germline genetic variation in cell cycle control genes on breast cancer survival: results from a population-based cohort. AB - INTRODUCTION: Somatic alterations have been shown to correlate with breast cancer prognosis and survival, but less is known about the effects of common inherited genetic variation. Of particular interest are genes involved in cell cycle pathways, which regulate cell division. METHODS: We examined associations between common germline genetic variation in 13 genes involved in cell cycle control (CCND1, CCND2, CCND3, CCNE1, CDK2 [p33], CDK4, CDK6, CDKN1A [p21, Cip1], CDKN1B [p27, Kip1], CDKN2A [p16], CDKN2B [p15], CDKN2C [p18], and CDKN2D [p19]) and survival among women diagnosed with invasive breast cancer participating in the SEARCH (Studies of Epidemiology and Risk factors in Cancer Heredity) breast cancer study. DNA from up to 4,470 women was genotyped for 85 polymorphisms that tag the known common polymorphisms (minor allele frequency > 0.05) in the genes. The genotypes of each polymorphism were tested for association with survival using Cox regression analysis. RESULTS: The rare allele of the tagging single nucleotide polymorphism (SNP) rs2479717 is associated with an increased risk of death (hazard ratio = 1.26 per rare allele carried, 95% confidence interval: 1.12 to 1.42; P = 0.0001), which was not attenuated after adjusting for tumour stage, grade, and treatment. This SNP is part of a large linkage disequilibrium block, which contains CCND3, BYSL, TRFP, USP49, C6ofr49, FRS3, and PGC. We evaluated the association of survival and somatic expression of these genes in breast tumours using expression microarray data from seven published datasets. Elevated expression of the C6orf49 transcript was associated with breast cancer survival, adding biological interest to the finding. CONCLUSION: It is possible that CCND3 rs2479717, or another variant it tags, is associated with prognosis after a diagnosis of breast cancer. Further study is required to validate this finding. PMID- 18507838 TI - Dynamic instability of the major urinary protein gene family revealed by genomic and phenotypic comparisons between C57 and 129 strain mice. AB - BACKGROUND: The major urinary proteins (MUPs) of Mus musculus domesticus are deposited in urine in large quantities, where they bind and release pheromones and also provide an individual 'recognition signal' via their phenotypic polymorphism. Whilst important information about MUP functionality has been gained in recent years, the gene cluster is poorly studied in terms of structure, genic polymorphism and evolution. RESULTS: We combine targeted sequencing, manual genome annotation and phylogenetic analysis to compare the Mup clusters of C57BL/6J and 129 strains of mice. We describe organizational heterogeneity within both clusters: a central array of cassettes containing Mup genes highly similar at the protein level, flanked by regions containing Mup genes displaying significantly elevated divergence. Observed genomic rearrangements in all regions have likely been mediated by endogenous retroviral elements. Mup loci with coding sequences that differ between the strains are identified--including a gene/pseudogene pair--suggesting that these inbred lineages exhibit variation that exists in wild populations. We have characterized the distinct MUP profiles in the urine of both strains by mass spectrometry. The total MUP phenotype data is reconciled with our genomic sequence data, matching all proteins identified in urine to annotated genes. CONCLUSION: Our observations indicate that the MUP phenotypic polymorphism observed in wild populations results from a combination of Mup gene turnover coupled with currently unidentified mechanisms regulating gene expression patterns. We propose that the structural heterogeneity described within the cluster reflects functional divergence within the Mup gene family. PMID- 18507839 TI - Squamosamide derivative FLZ protects dopaminergic neurons against inflammation mediated neurodegeneration through the inhibition of NADPH oxidase activity. AB - BACKGROUND: Inflammation plays an important role in the pathogenesis of Parkinson's disease (PD) through over-activation of microglia, which consequently causes the excessive production of proinflammatory and neurotoxic factors, and impacts surrounding neurons and eventually induces neurodegeneration. Hence, prevention of microglial over-activation has been shown to be a prime target for the development of therapeutic agents for inflammation-mediated neurodegenerative diseases. METHODS: For in vitro studies, mesencephalic neuron-glia cultures and reconstituted cultures were used to investigate the molecular mechanism by which FLZ, a squamosamide derivative, mediates anti-inflammatory and neuroprotective effects in both lipopolysaccharide-(LPS)- and 1-methyl-4-phenylpyridinium-(MPP+) mediated models of PD. For in vivo studies, a 1-methyl-4-phenyl-1, 2, 3, 6 tetrahydropyridine-(MPTP-) induced PD mouse model was used. RESULTS: FLZ showed potent efficacy in protecting dopaminergic (DA) neurons against LPS-induced neurotoxicity, as shown in rat and mouse primary mesencephalic neuronal-glial cultures by DA uptake and tyrosine hydroxylase (TH) immunohistochemical results. The neuroprotective effect of FLZ was attributed to a reduction in LPS-induced microglial production of proinflammatory factors such as superoxide, tumor necrosis factor-alpha (TNF-alpha), nitric oxide (NO) and prostaglandin E2 (PGE2). Mechanistic studies revealed that the anti-inflammatory properties of FLZ were mediated through inhibition of NADPH oxidase (PHOX), the key microglial superoxide-producing enzyme. A critical role for PHOX in FLZ-elicited neuroprotection was further supported by the findings that 1) FLZ's protective effect was reduced in cultures from PHOX-/- mice, and 2) FLZ inhibited LPS induced translocation of the cytosolic subunit of p47PHOX to the membrane and thus inhibited the activation of PHOX. The neuroprotective effect of FLZ demonstrated in primary neuronal-glial cultures was further substantiated by an in vivo study, which showed that FLZ significantly protected against MPTP-induced DA neuronal loss, microglial activation and behavioral changes. CONCLUSION: Taken together, our results clearly demonstrate that FLZ is effective in protecting against LPS- and MPTP-induced neurotoxicity, and the mechanism of this protection appears to be due, at least in part, to inhibition of PHOX activity and to prevention of microglial activation. PMID- 18507840 TI - Growing old at home - a randomized controlled trial to investigate the effectiveness and cost-effectiveness of preventive home visits to reduce nursing home admissions: study protocol [NCT00644826]. AB - BACKGROUND: Regarding demographic changes in Germany it can be assumed that the number of elderly and the resulting need for long term care is increasing in the near future. It is not only an individual's interest but also of public concern to avoid a nursing home admission. Current evidence indicates that preventive home visits can be an effective way to reduce the admission rate in this way making it possible for elderly people to stay longer at home than without home visits. As the effectiveness and cost-effectiveness of preventive home visits strongly depends on existing services in the social and health system existing international results cannot be merely transferred to Germany. Therefore it is necessary to investigate the effectiveness and cost-effectiveness of such an intervention in Germany by a randomized controlled trial. METHODS: The trial is designed as a prospective multi-center randomized controlled trial in the cities of Halle and Leipzig. The trial includes an intervention and a control group. The control group receives usual care. The intervention group receives three additional home visits by non-physician health professionals (1) geriatric assessment, (2) consultation, (3) booster session. The nursing home admission rate after 18 months will be defined as the primary outcome. An absolute risk reduction from a 20% in the control-group to a 7% admission rate in the intervention group including an assumed drop out rate of 30% resulted in a required sample size of N = 320 (n = 160 vs. n = 160). Parallel to the clinical outcome measurement the intervention will be evaluated economically. The economic evaluation will be performed from a society perspective. DISCUSSION: To the authors' knowledge for the first time a trial will investigate the effectiveness and cost-effectiveness of preventive home visits for people aged 80 and over in Germany using the design of a randomized controlled trial. Thus, the trial will contribute to the existing evidence on preventive home visits especially in Germany. PMID- 18507841 TI - SOCS3 inhibiting migration of A549 cells correlates with PYK2 signaling in vitro. AB - BACKGROUND: Suppressor of cytokine signaling 3 (SOCS3) is considered to inhibit cytokine responses and play a negative role in migration of various cells. Proline-rich tyrosine kinase 2 (PYK2) is a non-receptor kinase and has been found crucial to cell motility. However, little is known about whether SOCS3 could regulate PYK2 pro-migratory function in lung cancer. METHODS: The methylation status of SOCS3 was investigated in HBE and A549 cell lines by methylation specific PCR. A549 cells were either treated with a demethylation agent 5-aza-2' deoxycytidine or transfected with three SOCS3 mutants with various functional domains deleted. Besides, cells were pretreated with a proteasome inhibitor beta lactacystin where indicated. The effects of SOCS3 up-regulation on PYK2 expression, PYK2 and ERK1/2 phosphorylations were assessed by western blot using indicated antibodies. RT-PCR was used to estimate PYK2 mRNA levels. Transwell experiments were performed to evaluate cell migration. RESULTS: SOCS3 expression was found impaired in A549 cells and higher PYK2 activity was correlated with enhanced cell migration. We identified that SOCS3 was aberrantly methylated in the exon 2, and 5-aza-2'-deoxycytidine restored SOCS3 expression. Reactivation of SOCS3 attenuated PYK2 expression and phosphorylation, cell migration was inhibited as well. Transfection studies indicated that exogenous SOCS3 interacted with PYK2, and both the Src homology 2 (SH2) and the kinase inhibitory region (KIR) domains of SOCS3 contributed to PYK2 binding. Furthermore, SOCS3 was found to inhibit PYK2-associated ERK1/2 activity in A549 cells. SOCS3 possibly promoted degradation of PYK2 in a SOCS-box-dependent manner and interfered with PYK2 related signaling events, such as cell migration. CONCLUSION: These data indicate that SOCS3 negatively regulates cell motility and decreased SOCS3 induced by methylation may confer a migration advantage to A549 cells. These results also suggest a negative role of SOCS3 in PYK2 signaling, and a previously unidentified regulatory mechanism for PYK2 function. PMID- 18507842 TI - Humoral immunoreactivity to gliadin and to tissue transglutaminase is present in some patients with multiple myeloma. AB - BACKGROUND: Multiple myeloma (MM) is a clonal B-cell disorder with many immunological disturbances. The aim of this work was to assess whether some of food antigens contribute to the imbalance of immune response by screening the sera of MM patients for their immunoreactivity to food constituent gliadin, to tissue transglutaminase-2 (tTG-2) and to Ro/SSA antigen.Sera from 61 patients with MM in various stages of disease, before, or after some cycles of conventional therapy were analyzed by commercial Binding Site ELISA tests. The control group consisted of 50 healthy volunteers. Statistical analysis of data obtained was performed by Mann Whitney Test. RESULTS: The higher serum IgA immunoreactivity to gliadin was found in 14/56 patients and in one of control people. The enhanced serum IgG immunoreactivity to gliadin was found in only two of tested patients and in two controls. The enhanced IgA immunoreactivity to tTG 2 was found in 10/49 patients' sera, while 4/45 patients had higher serum IgG immunoreactivity. The enhanced serum IgG immunoreactivity to RoSSA antigen was found in 9/47 analyzed MM patients' sera. Statistical analysis of data obtained revealed that only the levels of anti-tTG-2 IgA immunoreactivity in patients with MM were significantly higher than these obtained in healthy controls (P < 0.02) CONCLUSION: Data obtained showed the existence of the enhanced serum immunoreactivity to gliadin, tTG-2 and Ro/SSA antigens in some patients with MM. These at least partially could contribute to the immunological imbalance frequently found in this disease. PMID- 18507843 TI - Absence of gastrointestinal infections in a cohort of patients with Zollinger Ellison syndrome and other acid hypersecretors receiving long-term acid suppression with lansoprazole. AB - BACKGROUND: The relationship between proton pump inhibitor therapy and other acid suppressing medications and the risk of gastrointestinal infections remains controversial. METHODS: Patients enrolled in a long-term trial of lansoprazole for Zollinger-Ellison syndrome and other acid hypersecretory states had interval histories taken every six months regarding hospitalizations or other intercurrent medical conditions. All medications taken were also reviewed at each visit. In addition, available patients were specifically queried during the study period 2006-2007 regarding the development of any gastrointestinal infections, hospitalizations, and prescriptions for antibiotics. RESULTS: Ninety patients were enrolled in our long-term study and 81 were available for review. The median basal gastric pH for the cohort after stabilization on therapy was 2.9 and ranged from 1.1 - 8.4 with a median pentagastrin stimulated gastric pH of 1.60 (range 1.0 - 8.2). No patient developed a clinically significant gastrointestinal infection during the study. The median patient years of follow-up were 6.25 years. CONCLUSION: In a cohort of patients with gastric acid hypersecretion in whom acid secretion status was monitored on lansoprazole, all were free of significant gastrointestinal infections on long-term follow-up. TRIAL REGISTRATION: NCT00204373. PMID- 18507844 TI - Experimental transmission of bovine spongiform encephalopathy to European red deer (Cervus elaphus elaphus). AB - BACKGROUND: Bovine spongiform encephalopathy (BSE), a member of the transmissible spongiform encephalopathies (TSE), primarily affects cattle. Transmission is via concentrate feed rations contaminated with infected meat and bone meal (MBM). In addition to cattle, other food animal species are susceptible to BSE and also pose a potential threat to human health as consumption of infected meat products is the cause of variant Creutzfeldt-Jakob disease in humans, which is invariably fatal. In the UK, farmed and free ranging deer were almost certainly exposed to BSE infected MBM in proprietary feeds prior to legislation banning its inclusion. Therefore, although BSE has never been diagnosed in any deer species, a possible risk to human health remains via ingestion of cervine products. Chronic wasting disease (CWD), also a TSE, naturally infects several cervid species in North America and is spreading rapidly in both captive and free-ranging populations. RESULTS: Here we show that European red deer (Cervus elaphus elaphus) are susceptible to intra-cerebral (i/c) challenge with BSE positive cattle brain pool material resulting in clinical neurological disease and weight loss by 794-1290 days and the clinical signs are indistinguishable to those reported in deer with CWD. Spongiform changes typical of TSE infections were present in brain and accumulation of the disease-associated abnormal prion protein (PrPd) was present in the central and peripheral nervous systems, but not in lymphoid or other tissues. Western immunoblot analysis of brain material showed a similar glycosylation pattern to that of BSE derived from infected cattle and experimentally infected sheep with respect to protease-resistant PrP isoforms. However, the di-, mono- and unglycosylated bands migrated significantly (p < 0.001) further in the samples from the clinically affected deer when compared to BSE infected brains of cattle and sheep. CONCLUSION: This study shows that deer are susceptible to BSE by intra-cerebral inoculation and display clinical signs and vacuolar pathology that are similar to those of CWD. These findings highlight the importance of preventing the spread to Europe of CWD from North America as this may necessitate even more extensive testing of animal tissues destined for human consumption within the EU. Although the absence of PrPd in lymphoid and other non-neurological tissues potentially limits the risk of transmission to humans, the replication of TSE agents in peripheral tissues following intra cerebral challenge is often limited. Thus the assessment of risk posed by cervine BSE as a human pathogen or for environmental contamination should await the outcome of ongoing oral challenge experiments. PMID- 18507845 TI - DREAM is reduced in synovial fibroblasts of patients with chronic arthritic pain: is it a suitable target for peripheral pain management? AB - INTRODUCTION: The endogenous pain-relieving system depends in part on the regulation of nociceptive signals through binding of opioids to the corresponding opioid receptor. Interfering with the trans-repression effect of downstream regulatory element antagonist modulator (DREAM) on the transcription of the opioid dynorphin-encoding prodynorphin (pdyn) gene might enhance pain relief in the periphery. METHODS: Expression levels were measured in osteoarthritis (OA) synovial fibroblast-like cells (SFLCs) (n = 8) and in peripheral blood mononuclear cells (PBMCs) from OA patients (n = 53) and healthy controls (n = 26) by real-time polymerase chain reaction. Lysed OA SFLCs were analyzed by immunoprecipitation. Translation of DREAM mRNA was inhibited by small interfering RNAs (siRNAs). Expressions of DREAM, pdyn, and c-fos mRNAs were measured at 24, 48, and 72 hours after transfection. RESULTS: The expression of DREAM mRNA was shown in both healthy and OA SFLCs as well as PBMCs. Inhibiting transcription using siRNAs led to a marked reduction in DREAM expression after 24, 48, and 72 hours. However, no significant changes in c-fos and pdyn expression occurred. In addition, DREAM mRNA expression was significantly reduced in OA patients with chronic pain (pain intensity as measured by a visual analog scale scale of greater than 40), but no pdyn expression was detectable. CONCLUSION: To our knowledge, this is the first report showing the expression of DREAM in SFLCs and PBMCs on the mRNA level. However, DREAM protein was not detectable. Since repression of pdyn transcription persists after inhibiting DREAM translation, DREAM appears to play no functional role in the kappa opioid receptor system in OA SFLCs. Therefore, our data suggest that DREAM appears not to qualify as a target in peripheral pain management. PMID- 18507846 TI - Regulation of survival in adult hippocampal and glioblastoma stem cell lineages by the homeodomain-only protein HOP. AB - BACKGROUND: Homeodomain proteins play critical roles in shaping the development of the embryonic central nervous system in mammals. After birth, neurogenic activities are relegated to stem cell niches, which include the subgranular layer of the dentate gyrus of the hippocampus. Here, we have analyzed the function of HOP (Homeodomain only protein) in this stem cell niche and in human glioblastomas. RESULTS: We find that HOP is strongly expressed by radial astrocytes of the dentate gyrus in mice, which are stem cells that give rise to hippocampal granular neurons throughout adulthood. Deletion or down-regulation of HOP results in a decrease of apoptosis of these stem cells without changes in proliferation, and in an increase in the number of newly formed granule neurons. We also find that human glioblastomas largely lack HOP expression and that reintroduction of HOP function in glioma cells cultured as gliomaspheres leads to enhanced apoptosis in a subset of cases. In these cells, HOP function decreases clonogenicity. CONCLUSION: These data suggest that HOP participates in the regulation of the adult mouse hippocampal stem cell niche by negatively affecting cell survival. In addition, HOP may work as a tumor suppressor in a subset of glioblastomas. HOP function thus appears to be critical in the adult brain in a region of continued plasticity, and its deregulation may contribute to disease. PMID- 18507847 TI - Global impression of perceived difficulties in children and adolescents with attention-deficit/hyperactivity disorder: reliability and validity of a new instrument assessing perceived difficulties from a patient, parent and physician perspective over the day. AB - BACKGROUND: The objective of this analysis was to evaluate the psychometric properties of a brief scale developed to assess the degree of difficulties in children with Attention-Deficit/Hyperactivity Disorder (ADHD). The Global Impression of Perceived Difficulties (GIPD) scale reflects overall impairment, psychosocial functioning and Quality of Life (QoL) as rated by patient, parents and physician at various times of the day. METHODS: In two open-label studies, ADHD-patients aged 6-17 years were treated with atomoxetine (target-dose 0.5-1.2 mg/kg/day). ADHD-related difficulties were assessed up to week 24 using the GIPD. Data from both studies were combined to validate the scale. RESULTS: Overall, 421 patients received atomoxetine. GIPD scores improved over time. All three GIPD versions (patient, parent, physician) were internally consistent; all items showed at least moderate item-total correlation. The scale showed good test retest reliability over a two-week period from all three perspectives. Good convergent and discriminant validity was shown. CONCLUSION: GIPD is an internally consistent, reliable and valid measure to assess difficulties in children with ADHD at various times of the day and can be used as indicator for psychosocial impairment and QoL. The scale is sensitive to treatment-related change. PMID- 18507848 TI - Emotional well-being in children and adolescents treated with atomoxetine for attention-deficit/hyperactivity disorder: findings from a patient, parent and physician perspective using items from the pediatric adverse event rating scale (PAERS). AB - BACKGROUND: The objective of this analysis was to measure changes in items on the Pediatric Adverse Event Rating Scale (PAERS) that relate to emotional well-being of children and adolescents with Attention-Deficit/Hyperactivity Disorder (ADHD) during treatment with atomoxetine for up to 24 weeks from the perspective of the patient, the parent, and the physician. METHODS: Patients aged 6-17 years with ADHD were treated with atomoxetine (target dose 1.2 mg/kg/day). In the two studies on which this secondary analysis is based the PAERS was used to assess the tolerability of atomoxetine in children and adolescents. This scale has a total of 48 items. The ten items that reflect emotional well-being were selected to measure changes over time from a patient, parent, and physician perspective. RESULTS: 421 patients were treated with atomoxetine. 355 patients completed the 8 week treatment period, and 260 patients completed the 24-week treatment period. The ten items that reflect emotional well-being were grouped in five dimensions: depressed mood, self-harm, irritability/agitation, drowsiness, and euphoria. The scores of these dimensions decreased over time, both from a patient as well as from a parent and physician perspective. Only the dimension self-harm was extremely low at baseline and stayed low over time. The mean scores for the ten items depended on the rater perspective. CONCLUSION: The emotional well-being of children and adolescents with ADHD improved in terms of depressed mood, irritability/agitation, drowsiness, and euphoria during treatment with atomoxetine for up to 24 weeks. PMID- 18507849 TI - Measurement of left ventricular dimensions with contrast-enhanced three dimensional cine imaging facilitated by k-t SENSE. AB - AIM: To compare three-dimensional (3D) k-t sensitivity encoded (k-t SENSE) cine cardiovascular magnetic resonance (CMR), before and after contrast administration, against standard 2D imaging for the assessment of left ventricular volumes and mass. METHOD: Twenty-six subjects (14 volunteers, 12 patients) underwent multiple breathhold 2D balanced turbo-field echo cine CMR in addition to k-t SENSE accelerated 3D imaging (acceleration factor 5; 5x k-t SENSE), performed before and after administration of a high-relaxivity gadolinium based contrast agent (Gadobutrolum). k-t acceleration factors of 7 and 10 were also assessed in six volunteers. Left ventricular end diastolic volume (EDV), end systolic volume (ESV), mass, and ejection fraction (EF) were calculated for each method. RESULTS: There was at least moderate agreement between the EDV, ESV, mass and EF calculated by 2D and 3D 5x k-t SENSE methods before contrast (concordance coefficients 0.92, 0.95, 0.97, 0.92, respectively). Agreement improved following contrast (concordance coefficients 0.97, 0.99, 0.98, 0.93, respectively). The 3D method underestimated all parameters compared to 2D (mean bias pre-contrast 6.1 ml, 0.6 ml, 3.5 g, 2.0% respectively). 3D image quality scores were significantly poorer than 2D, showing a non-significant trend to improvement following contrast administration. Parameters derived with k-t acceleration factors of 7 and 10 showed poorer agreement with 2D values. CONCLUSION: Left ventricular volumes and mass are reliably assessed using 3D 5x k-t SENSE accelerated CMR. Contrast administration further improves agreement between 5x k-t SENSE and 2D-derived measurements. k-t acceleration factors greater than 5, though feasible, produce poorer agreement with 2D values. PMID- 18507850 TI - Exploring general practitioners' experience of informing women about prenatal screening tests for foetal abnormalities: a qualitative focus group study. AB - BACKGROUND: Recent developments have made screening tests for foetal abnormalities available earlier in pregnancy and women have a range of testing options accessible to them. It is now recommended that all women, regardless of their age, are provided with information on prenatal screening tests. General Practitioners (GPs) are often the first health professionals a woman consults in pregnancy. As such, GPs are well positioned to inform women of the increasing range of prenatal screening tests available. The aim of this study was to explore GPs experience of informing women of prenatal genetic screening tests for foetal abnormality. METHODS: A qualitative study consisting of four focus groups was conducted in metropolitan and rural Victoria, Australia. A discussion guide was used and the audio-taped transcripts were independently coded by two researchers using thematic analysis. Multiple coders and analysts and informant feedback were employed to reduce the potential for researcher bias and increase the validity of the findings. RESULTS: Six themes were identified and classified as 'intrinsic' if they occurred within the context of the consultation or 'extrinsic' if they consisted of elements that impacted on the GP beyond the scope of the consultation. The three intrinsic themes were the way GPs explained the limitations of screening, the extent to which GPs provided information selectively and the time pressures at play. The three extrinsic factors were GPs' attitudes and values towards screening, the conflict they experienced in offering screening information and the sense of powerlessness within the screening test process and the health care system generally. Extrinsic themes reveal GPs' attitudes and values to screening and to disability, as well as raising questions about the fundamental premise of testing. CONCLUSION: The increasing availability and utilisation of screening tests, in particular first trimester tests, has expanded GPs' role in facilitating women's informed decision-making. Recognition of the importance of providing this complex information warrants longer consultations to respond to the time pressures that GPs experience. Understanding the intrinsic and extrinsic factors that impact on GPs may serve to shape educational resources to be more appropriate, relevant and supportive. PMID- 18507851 TI - Stabilization of the genome of the mismatch repair deficient Mycobacterium tuberculosis by context-dependent codon choice. AB - BACKGROUND: The rate at which a stretch of DNA mutates is determined by the cellular systems for DNA replication and repair, and by the nucleotide sequence of the stretch itself. One sequence feature with a particularly strong influence on the mutation rate are nucleotide repeats. Some microbial pathogens use nucleotide repeats in their genome to stochastically vary phenotypic traits and thereby evade host defense. However, such unstable sequences also come at a cost, as mutations are often deleterious. Here, we analyzed how these opposing forces shaped genome stability in the human pathogen Mycobacterium tuberculosis. M. tuberculosis lacks a mismatch repair system, and this renders nucleotide repeats particularly unstable. RESULTS: We found that proteins of M. tuberculosis are encoded by using codons in a context-dependent manner that prevents the emergence of nucleotide repeats. This context-dependent codon choice leads to a strong decrease in the estimated frame-shift mutation rate and thus to an increase in genome stability. CONCLUSION: These results indicate that a context-specific codon choice can partially compensate for the lack of a mismatch repair system, and helps to maintain genome integrity in this pathogen. PMID- 18507852 TI - Strategies to improve global influenza surveillance: a decision tool for policymakers. AB - BACKGROUND: Global pandemic influenza preparedness relies heavily on public health surveillance, but it is unclear that current surveillance fully meets pandemic preparedness needs. METHODS: We first developed a conceptual framework to help systematically identify strategies to improve the detection of an early case or cluster of novel human influenza disease during the pre-pandemic period. We then developed a process model (flow diagram) depicting nine major pathways through which a case in the community could be detected and confirmed, and mapped the improvement strategies onto this model. Finally, we developed an interactive decision tool by building quantitative measures of probability and time into each step of the process model and programming it to calculate the net probability and time required for case detection through each detection pathway. Input values for each step can be varied by users to assess the effects of different improvement strategies, alone or in combination. We illustrate application of the tool using hypothetical input data reflecting baseline and 12-month follow-up scenarios, following concurrent implementation of multiple improvement strategies. RESULTS: We compared outputs from the tool across detection pathways and across time, at baseline and 12-month follow up. The process model and outputs from the tool suggest that traditional efforts to build epidemiology and laboratory capacity are efficient strategies, as are more focused strategies within these, such as targeted laboratory testing; expedited specimen transport; use of technologies to streamline data flow; and improved reporting compliance. Other promising strategies stem from community detection - better harnessing of electronic data mining and establishment of community-based monitoring. CONCLUSION: Our practical tool allows policymakers to use their own realistic baseline values and program projections to assess the relative impact of different interventions to improve the probability and timeliness of detecting early human cases or clusters caused by a novel influenza virus, a possible harbinger of a new pandemic. Policymakers can use results to target investments to improve their surveillance infrastructure. Multi-national planners can also use the tool to help guide directions in surveillance system improvements more globally. Finally, our systematic approach can also be tailored to help improve surveillance for other diseases. PMID- 18507853 TI - Trends in prevalence of substance use among Icelandic adolescents, 1995-2006. AB - BACKGROUND: Adolescent substance use continues to be of great global public health concern in many countries with advanced economies. Previous research has shown that substance use among 15-16 year-old-youth has increased in many European countries in recent years. The aim of this study was to examine trends in prevalence of daily smoking, alcohol intoxication, and illicit substance use among Icelandic adolescents. METHODS: Repeated-measures, population-based cross sectional surveys of between 3,100 and 3,900 10th-grade students who participated in the annual Youth of Iceland studies were analyzed, with response rates of between 80% and 90%. RESULTS: The prevalence of daily smoking, alcohol intoxication, and illicit substance use was at a peak in 1998, with almost 23% having reported daily smoking, 42% having reported becoming intoxicated at least once during the last 30 days, and over 17% having used hashish once or more often in their lifetime. By 2006, daily smoking had declined to 12%, having become intoxicated once or more often during the last 30 days to 25%, and having ever used hashish declined to 9%. CONCLUSION: The prevalence of substance use among Icelandic 10th graders declined substantially from 1995 to 2006. Proportions of adolescents who smoke cigarettes, had become intoxicated during the last 30 days, as well as those admitting to hashish use all decreased to a great deal during the period under study. The decline in prevalence of adolescent substance use in Iceland is plausibly the result of local community collaboration where researchers, policy makers and practitioners who work with young people have combined their efforts. PMID- 18507854 TI - Analysis of the effects of exposure to acute hypoxia on oxidative lesions and tumour progression in a transgenic mouse breast cancer model. AB - BACKGROUND: Tumour hypoxia is known to be a poor prognostic indicator, predictive of increased risk of metastatic disease and reduced survival. Genomic instability has been proposed as one of the potential mechanisms for hypoxic tumour progression. Both of these features are commonly found in many cancer types, but their relationship and association with tumour progression has not been examined in the same model. METHODS: To address this issue, we determined the effects of 6 week in vivo acute hypoxic exposure on the levels of mutagenic lipid peroxidation product, malondialdehyde, and 8-oxo-7,8-dihydro-2'-deoxyguanosine DNA (8-oxo-dG) lesions in the transgenic polyomavirus middle T (PyMT) breast cancer mouse model. RESULTS: We observed significantly increased plasma lipid peroxidation and 8-oxo dG lesion levels in the hypoxia-exposed mice. Consumption of malondialdehyde also induced a significant increase in the PyMT tumour DNA lesion levels, however, these increases did not translate into enhanced tumour progression. We further showed that the in vivo exposure to acute hypoxia induced accumulation of F4/80 positive tumour-associated macrophages (TAMs), demonstrating a relationship between hypoxia and macrophages in an experimental model. CONCLUSION: These data suggest that although exposure to acute hypoxia causes an increase in 8-oxo-dG lesions and TAMs in the PyMT tumours, these increases do not translate into significant changes in tumour progression at the primary or metastatic levels in this strong viral oncogene-driven breast cancer model. PMID- 18507855 TI - Reproductive toxicity of seafood contaminants: prospective comparisons of Swedish east and west coast fishermen's families. AB - Cohorts comprising fishermen's families on the east coast of Sweden have been found to have a high consumption of contaminated fish as well as high body burdens of persistent organochlorine pollutants (POPs). Their west coast correspondents are socio-economically similar, but with considerably lower POP exposure since the fish caught on the west coast is far less contaminated. The rationale for this was that the cohorts residing on the east coast of Sweden have been found to have a high consumption of contaminated fish as well as high body burdens of POPs, whereas their west coast correspondents are socio-economically similar, but with considerably lower POP exposure since the fish caught on the west coast is far less contaminated. Among the reproductive outcomes investigated are included both male and female parameters, as well as couple fertility and effects on the fetus. A range of exposure measures, including both questionnaire assessments of fish consumption and biomarkers, have been used. The most consistent findings of the studies are those related to the fetus, where a decreased birth weight was found across all measures of exposure, which is in agreement with studies from other populations. Some markers for male reproduction function, i.e. sperm motility, sperm chromatin integrity, and Y:X chromosome ratio, were associated with POP exposure, whereas others, such as sperm concentration and semen volume, were not. With respect to couple fertility and female reproductive parameters, no support was given for associations with POP exposure. Although some associations may have been affected by beneficial effects of essential nutrients in seafood, the overall findings are meaningful in the context of reproductive toxicity and support the usefulness of the epidemiological design. PMID- 18507856 TI - TileQC: a system for tile-based quality control of Solexa data. AB - BACKGROUND: Next-generation DNA sequencing technologies such as Illumina's Solexa platform and Roche's 454 approach provide new avenues for investigating genome scale questions. However, they also present novel analytical challenges that must be met for their effective application to biological questions. RESULTS: Here we report the availability of tileQC, a tile-based quality control system for Solexa data written in the R language. TileQC provides a means of recognizing bias and error in Solexa output by graphically representing data generated by flow cell tiles. The data represented in the images is then made available in the R environment for further analysis and automation of error detection. CONCLUSION: TileQC offers a highly adaptable and powerful tool for the quality control of Solexa-based DNA sequence data. PMID- 18507857 TI - Altered splicing of CEACAM1 in breast cancer: identification of regulatory sequences that control splicing of CEACAM1 into long or short cytoplasmic domain isoforms. AB - BACKGROUND: Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), a cell adhesion molecule expressed in a variety of cell types is a putative tumor suppressor gene. Alternative splicing of CEACAM1 generates 11 different splice variants, which include 1-4 ectodomains with either short or long cytoplasmic domain generated by the exclusion (CEACAM1-S) or inclusion (CEACAM1-L) of exon 7. Studies in rodents indicate that optimal ratios of CEACAM1 splice variants are required to inhibit colonic tumor cell growth. RESULTS: We show that CEACAM1 is expressed in a tissue specific manner with significant differences in the ratios of its short (CEACAM1-S) and long (CEACAM1-L) cytoplasmic domain splice variants. Importantly, we find dramatic differences between the ratios of S:L isoforms in normal breast tissues versus breast cancer specimens, suggesting that altered splicing of CEACAM1 may play an important role in tumorogenesis. Furthermore, we have identified two regulatory cis-acting elements required for the alternative splicing of CEACAM1. Replacement of these regulatory elements by human beta globin exon sequences resulted in exon 7-skipped mRNA as the predominant product. Interestingly, while insertion of exon 7 in a beta-globin reporter gene resulted in its skipping, exon 7 along with the flanking intron sequences recapitulated the alternative splicing of CEACAM1. CONCLUSION: Our results indicate that a network of regulatory elements control the alternative splicing of CEACAM1. These findings may have important implications in therapeutic modalities of CEACAM1 linked human diseases. PMID- 18507858 TI - Guyon tunnel syndrome secondary to excessive healing tissue in a child: a case report. AB - We describe a case of an 8-year-old boy who developed a combined motor and sensory neuropathy of the distal ulnar nerve, after sustaining a superficial injury to the right flexor carpi ulnaris tendon at the level of the distal wrist crease. Guyon's canal syndrome is a very rare entity during childhood. We have noted only one prior description of this syndrome in the pediatric age group in a review of the English literature. PMID- 18507859 TI - Expression of novel extracellular sulfatases Sulf-1 and Sulf-2 in normal and osteoarthritic articular cartilage. AB - INTRODUCTION: Changes in sulfation of cartilage glycosaminoglycans as mediated by sulfatases can regulate growth factor signaling. The aim of this study was to analyze expression patterns of recently identified extracellular sulfatases Sulf 1 and Sulf-2 in articular cartilage and chondrocytes. METHODS: Sulf-1 and Sulf-2 expressions in human articular cartilage from normal donors and patients with osteoarthritis (OA) and in normal and aged mouse joints were analyzed by real time polymerase chain reaction, immunohistochemistry, and Western blotting. RESULTS: In normal articular cartilage, Sulf-1 and Sulf-2 mRNAs and proteins were expressed predominantly in the superficial zone. OA cartilage showed significantly higher Sulf-1 and Sulf-2 mRNA expression as compared with normal human articular cartilage. Sulf protein expression in OA cartilage was prominent in the cell clusters. Western blotting revealed a profound increase in Sulf protein levels in human OA cartilage. In normal mouse joints, Sulf expression was similar to human cartilage, and with increasing age, there was a marked upregulation of Sulf. CONCLUSION: The results show low levels of Sulf expression, restricted to the superficial zone in normal articular cartilage. Sulf mRNA and protein levels are increased in aging and OA cartilage. This increased Sulf expression may change the sulfation patterns of heparan sulfate proteoglycans and growth factor activities and thus contribute to abnormal chondrocyte activation and cartilage degradation in OA. PMID- 18507861 TI - Reproducibility and geometric accuracy of the Fixster system during hypofractionated stereotactic radiotherapy. AB - BACKGROUND: Hypofractionated radiotherapy has been used for the treatment of AVMs and brain metastases. Hypofractionation necessitates the use of a relocatable stereotactic frame that has to be applied on several occasions. The stereotactic frame needs to have a high degree of reproducibility, and patient positioning is crucial to achieve a high accuracy of the treatment. METHODS: In this study we have, by radiological means, evaluated the reproducibility of the isocenter in consecutive treatment sessions using the Fixster frame. Deviations in the X, Y and Z-axis were measured in 10 patients treated with hypofractionated radiotherapy. RESULTS: The mean deviation in the X-axis was 0.4 mm (range -2.1 - 2.1, median 0.7 mm) and in the Y-axis -0.3 mm (range -1.4 - 0.7, median -0.2 mm). The mean deviation in the Z-axis was -0.6 (range -1.4 - 1.4, median 0.0 mm). CONCLUSION: There is a high degree of reproducibility of the isocenter during successive treatment sessions with HCSRT using the Fixster frame for stereotactic targeting. The high reducibility enables a safe treatment using hypofractionated stereotactic radiotherapy. PMID- 18507860 TI - Characterization of three new serous epithelial ovarian cancer cell lines. AB - BACKGROUND: Cell lines constitute a powerful model to study cancer, and here we describe three new epithelial ovarian cancer (EOC) cell lines derived from poorly differentiated serous solid tumors (TOV-1946, and TOV-2223G), as well as the matched ascites for one case (OV-1946). METHODS: In addition to growth parameters, the cell lines were characterized for anchorage independent growth, migration and invasion potential, ability to form spheroids and xenografts in SCID mice. RESULTS: While all cell lines were capable of anchorage independent growth, only the TOV-1946 and OV-1946 cell lines were able to form spheroid and produce tumors. Profiling of keratins, p53 and Her2 protein expression was assessed by immunohistochemistry and western blot analyses. Somatic TP53 mutations were found in all cell lines, with TOV-1946 and OV-1946 harboring the same mutation, and none harbored the commonly observed somatic mutations in BRAF, KRAS or germline BRCA1/2 mutations found to recur in the French Canadian population. Conventional cytogenetics and spectral karyotype (SKY) analyses revealed complex karyotypes often observed in ovarian disease. CONCLUSION: This is the first report of the establishment of matched EOC cell lines derived from both solid tumor and ascites of the same patient. PMID- 18507862 TI - Elevated serum levels of soluble CD154 in children with juvenile idiopathic arthritis. AB - OBJECTIVE: Cytokines play important roles in mediating inflammation in autoimmunity. Several cytokines are elevated in serum and synovial fluid samples from children with Juvenile Idiopathic Arthritis (JIA). Soluble CD154 (sCD154) is elevated in other autoimmune disorders, but has not been characterized in JIA. Our objectives were to determine if sCD154 is elevated in JIA, and to examine correlations between sCD154 and other inflammatory cytokines. METHODS: Serum from 77 children with JIA and 81 pediatric controls was analyzed for interleukin (IL)1beta, IL2, IL4, IL5, IL6, IL8, IL10, IL12, IL13, sCD154, interferon-gamma (IFNgamma), soluble IL2 receptor (sIL2R), and tumor necrosis factor-alpha (TNFalpha), using the Luminex Multi-Analyte Profiling system. Differences in levels of cytokines between cases and controls were analyzed. Logistic regression was also performed. RESULTS: sCD154 was significantly elevated in cases compared to controls (p < 0.0001). IL1beta, IL5, IL6, IL8, IL13, IFNgamma, sIL2R, and TNFalpha were also significantly elevated in JIA. Levels of sCD154 were highly correlated with IL1beta, IL6, IL8, and TNFalpha (p < 0.0001). Logistic regression analysis suggested that IL6 (odds ratio (OR): 1.4, p < 0.0001), sCD154 (OR: 1.1, p < 0.0001), and TNFalpha (OR: 1.1, p < 0.005) were positively associated with JIA, while IL10 (OR: 0.5, p < 0.002) was protective. sCD154 was elevated in all JIA subtypes, with highest levels among more severe subtypes. IL1beta, IL6, IL8, sIL2R and TNFalpha were also elevated in several JIA subtypes. CONCLUSION: Serum levels of sCD154, IL1beta, IL6, IL8, sIL2R and TNFalpha are elevated in most JIA subtypes, suggesting a major role for sCD154, and these cytokines and cytokine receptors in the pathogenesis of JIA. PMID- 18507863 TI - A procedure to characterize geographic distributions of rare disorders in cohorts. AB - BACKGROUND: Individual point data can be analyzed against an entire cohort instead of only sampled controls to accurately picture the geographic distribution of populations at risk for low prevalence diseases. Analyzed as individual points, many smaller clusters with high relative risks (RR) and low empirical p values are indistinguishable from a random distribution. When points are aggregated into areal units, small clusters may result in a larger cluster with a low RR or be lost if divided into pieces included in units of larger populations that show no increased prevalence. Previous simulation studies showed lowered validity of spatial scan tests for true clusters with low RR. Using simulations, this study explored the effects of low cluster RR and areal unit size on local area clustering test (LACT) results, proposing a procedure to improve accuracy of cohort spatial analysis for rare events. RESULTS: Our simulations demonstrated the relationship of true RR to observed RR and p values with various, randomly located, cluster shapes, areal unit sizes and scanning window shapes in a diverse population distribution. Clusters with RR < 1.7 had elevated observed RRs and high p values. We propose a cluster identification procedure that applies parallel multiple LACTs, one on point data and three on two distinct sets of areal units created with varying population parameters that minimize the range of population sizes among units. By accepting only clusters identified by all LACTs, having a minimum population size, a minimum relative risk and a maximum p value, this procedure improves the specificity achieved by any one of these tests alone on a cohort study of low prevalence data while retaining sensitivity for small clusters. The procedure is demonstrated on two study regions, each with a five-year cohort of births and cases of a rare developmental disorder. CONCLUSION: For truly exploratory research on a rare disorder, false positive clusters can cause costly diverted research efforts. By limiting false positives, this procedure identifies 'crude' clusters that can then be analyzed for known demographic risk factors to focus exploration for geographically-based environmental exposure on areas of otherwise unexplained raised incidence. PMID- 18507864 TI - Measurement properties of the Inventory of Cognitive Bias in Medicine (ICBM). AB - BACKGROUND: Understanding how doctors think may inform both undergraduate and postgraduate medical education. Developing such an understanding requires valid and reliable measurement tools. We examined the measurement properties of the Inventory of Cognitive Bias in Medicine (ICBM), designed to tap this domain with specific reference to medicine, but with previously questionable measurement properties. METHODS: First year postgraduate entry medical students at Flinders University, and trainees (postgraduate doctors in any specialty) and consultants (N = 348) based at two teaching hospitals in Adelaide, Australia, completed the ICBM and a questionnaire measuring thinking styles (Rational Experiential Inventory). RESULTS: Questions with the lowest item-total correlation were deleted from the original 22 item ICBM, although the resultant 17 item scale only marginally improved internal consistency (Cronbach's alpha = 0.61 compared with 0.57). A factor analysis identified two scales, both achieving only alpha = 0.58. Construct validity was assessed by correlating Rational Experiential Inventory scores with the ICBM, with some positive correlations noted for students only, suggesting that those who are naive to the knowledge base required to "successfully" respond to the ICBM may profit by a thinking style in tune with logical reasoning. CONCLUSION: The ICBM failed to demonstrate adequate content validity, internal consistency and construct validity. It is unlikely that improvements can be achieved without considered attention to both the audience for which it is designed and its item content. The latter may need to involve both removal of some items deemed to measure multiple biases and the addition of new items in the attempt to survey the range of biases that may compromise medical decision making. PMID- 18507866 TI - Australia's insurance crisis and the inequitable treatment of self-employed midwives. AB - Based upon a review of articles published in Australia's major newspapers over the period January 2001 to December 2005, a case study approach has been used to investigate why, when compared with other small business operators, including medical specialists, Australian governments have appeared reluctant to protect the economic viability of the businesses of self-employed midwives. Theories of agenda setting and structuralism have been used to explore that inequity. What has emerged is a picture of the complex of factors that may have operated, and may be continuing to operate, to shape the policy agenda and thus prevent solutions to the insurance problems of self-employed midwives being found. PMID- 18507865 TI - Oil palm (Elaeis guineensis Jacq.) tissue culture ESTs: identifying genes associated with callogenesis and embryogenesis. AB - BACKGROUND: Oil palm (Elaeis guineensis Jacq.) is one of the most important oil bearing crops in the world. However, genetic improvement of oil palm through conventional breeding is extremely slow and costly, as the breeding cycle can take up to 10 years. This has brought about interest in vegetative propagation of oil palm. Since the introduction of oil palm tissue culture in the 1970s, clonal propagation has proven to be useful, not only in producing uniform planting materials, but also in the development of the genetic engineering programme. Despite considerable progress in improving the tissue culture techniques, the callusing and embryogenesis rates from proliferating callus cultures remain very low. Thus, understanding the gene diversity and expression profiles in oil palm tissue culture is critical in increasing the efficiency of these processes. RESULTS: A total of 12 standard cDNA libraries, representing three main developmental stages in oil palm tissue culture, were generated in this study. Random sequencing of clones from these cDNA libraries generated 17,599 expressed sequence tags (ESTs). The ESTs were analysed, annotated and assembled to generate 9,584 putative unigenes distributed in 3,268 consensi and 6,316 singletons. These unigenes were assigned putative functions based on similarity and gene ontology annotations. Cluster analysis, which surveyed the relatedness of each library based on the abundance of ESTs in each consensus, revealed that lipid transfer proteins were highly expressed in embryogenic tissues. A glutathione S transferase was found to be highly expressed in non-embryogenic callus. Further analysis of the unigenes identified 648 non-redundant simple sequence repeats and 211 putative full-length open reading frames. CONCLUSION: This study has provided an overview of genes expressed during oil palm tissue culture. Candidate genes with expression that are modulated during tissue culture were identified. However, in order to confirm whether these genes are suitable as early markers for embryogenesis, the genes need to be tested on earlier stages of tissue culture and a wider range of genotypes. This collection of ESTs is an important resource for genetic and genome analyses of the oil palm, particularly during tissue culture development. PMID- 18507867 TI - The global workforce shortages and the migration of medical professions: the Australian policy response. AB - Medical migration sees the providers of medical services (in particular medical practitioners) moving from one region or country to another. This creates problems for the provision of public health and medical services and poses challenges for laws in the nation state and for laws in the global community.There exists a global shortage of healthcare professionals. Nation states and health rights movements have been both responsible for, and responsive to, this global community shortage through a variety of health policy, regulation and legislation which directly affects the migration of medical providers. The microcosm responses adopted by individual nation states, such as Australia, to this workforce shortage further impact on the global workforce shortage through active recruitment of overseas-trained healthcare professionals. "Push" and "pull" factors exist which encourage medical migration of healthcare professionals. A nation state's approach to health policy, regulation and legislation dramatically helps to create these "push factors" and "pull factors". A co-ordinated global response is required with individual nation states being cognisant of the impact of their health policy, regulations and legislation on the global community through the medical migration of healthcare professionals. PMID- 18507868 TI - Prospective randomized study for optimal insulin therapy in type 2 diabetic patients with secondary failure. AB - BACKGROUND: The large clinical trials proved that Basal-Bolus (BB) insulin therapy was effective in the prevention of diabetic complications and their progression. However, BB therapy needs multiple insulin injections per a day. In this regard, a biphasic insulin analogue needs only twice-daily injections, and is able to correct postprandial hyperglycemia. Therefore it may achieve the blood glucose control as same as that of BB therapy and prevent the diabetic complications including macroangiopathy. METHODS: In PROBE (Prospective, Randomized, Open, Blinded-Endpoint) design, forty-two type 2 diabetic patients (male: 73.8%, median(inter quartile range) age: 64.5(56.8-71.0)years) with secondary failure of sulfonylurea (SU) were randomly assigned to BB therapy with a thrice-daily insulin aspart and once-daily basal insulin (BB group) or to conventional therapy with a twice-daily biphasic insulin analogue (30 Mix group), and were followed up for 6 months to compare changes in HbA1c, daily glycemic profile, intima-media thickness (IMT) of carotid artery, adiponectin levels, amounts of insulin used, and QOL between the two groups. RESULTS: After 6 months, HbA1c was significantly reduced in both groups compared to baseline (30 Mix; 9.3(8.1-11.3) --> 7.4(6.9-8.7)%, p < 0.01, vs BB;8.9(7.7-10.0) --> 6.9(6.2-7.3)%, p < 0.01), with no significant difference between the groups in percentage change in HbA1c (30 Mix; -14.7(-32.5- (-)7.5)% vs BB -17.8(-30.1- (-)11.1)%, p = 0.32). There was a significant decrease in daily glycemic profile at all points except dinner time in both groups compared to baseline. There was a significant increase in the amount of insulin used in the 30 Mix group after treatment compared to baseline (30 Mix;0.30(0.17-0.44) --> 0.39(0.31-0.42) IU/kg, p = 0.01). There was no significant difference in IMT, BMI, QOL or adiponectin levels in either group compared to baseline. CONCLUSION: Both BB and 30 mix group produced comparable reductions in HbA1c in type 2 diabetic patients with secondary failure. There was no significant change in IMT as an indicator of early atherosclerotic changes between the two groups. The basal-bolus insulin therapy may not be necessarily needed if the type 2 diabetic patients have become secondary failure. TRIAL REGISTRATION: Current Controlled Trials number, NCT00348231. PMID- 18507869 TI - Epigenetic modification of histone 3 at lysine 9 in sheep zygotes and its relationship with DNA methylation. AB - BACKGROUND: Previous studies indicated that, unlike mouse zygotes, sheep zygotes lacked the paternal DNA demethylation event. Another epigenetic mark, histone modification, especially at lysine 9 of histone 3 (H3K9), has been suggested to be mechanically linked to DNA methylation. In mouse zygotes, the absence of methylated H3K9 from the paternal pronucleus has been thought to attribute to the paternal DNA demethylation. RESULTS: By using the immunofluorescence staining approach, we show that, despite the difference in DNA methylation, modification of H3K9 is similar between the sheep and mouse zygotes. In both species, H3K9 is hyperacetylated or hypomethylated in paternal pronucleus relative to maternal pronucleus. In fact, sheep zygotes can also undergo paternal DNA demethylation, although to a less extent than the mouse. Further examinations of individual zygotes by double immunostaining revealed that, the paternal levels of DNA methylation were not closely associated with that of H3K9 acetylation or tri methylation. Treatment of either 5-azacytidine or Trichostatin A did not induce a significant decrease of paternal DNA methylation levels. CONCLUSION: Our results suggest that in sheep lower DNA demethylation of paternal genomes is not due to the H3K9 modification and the methylated DNA sustaining in paternal pronucleus does not come from DNA de novo methylation. PMID- 18507870 TI - JNK pathway is involved in the inhibition of inflammatory target gene expression and NF-kappaB activation by melittin. AB - BACKGROUND: Bee venom therapy has been used to treat inflammatory diseases including rheumatoid arthritis in humans and in experimental animals. We previously found that bee venom and melittin (a major component of bee venom) have anti-inflammatory effect by reacting with the sulfhydryl group of p50 of nuclear factor-kappa B (NF-kappaB) and IkappaB kinases (IKKs). Since mitogen activated protein (MAP) kinase family is implicated in the NF-kappaB activation and inflammatory reaction, we further investigated whether activation of MAP kinase may be also involved in the anti-inflammatory effect of melittin and bee venom. METHODS: The anti-inflammatory effects of melittin and bee venom were investigated in cultured Raw 264.7 cells, THP-1 human monocytic cells and Synoviocytes. The activation of NF-kappaB was investigated by electrophoretic mobility shift assay. Nitric oxide (NO) and prostaglandin E2 (PGE2) were determined either by Enzyme Linked Immuno Sorbent Assay or by biochemical assay. Expression of IkappaB, p50, p65, inducible nitric oxide synthetase (iNOS), cyclooxygenase-2 (COX-2) as well as phosphorylation of MAP kinase family was determined by Western blot. RESULTS: Melittin (0.5-5 mug/ml) and bee venom (5 and 10 mug/ml) inhibited lipopolysaccharide (LPS, 1 mug/ml) and sodium nitroprusside (SNP, 200 muM)-induced activation of c-Jun NH2-terminal kinase (JNK) in RAW 264.7 cells in a dose dependent manner. However, JNK inhibitor, anthra [1,9-cd]pyrazole 6 (2H)-one (SP600215, 10-50 muM) dose dependently suppressed the inhibitory effects of melittin and bee venom on NF-kappaB dependent luciferase and DNA binding activity via suppression of the inhibitory effect of melittin and bee venom on the LPS and SNP-induced translocation of p65 and p50 into nucleus as well as cytosolic release of IkappaB. Moreover, JNK inhibitor suppressed the inhibitory effects of melittin and bee venom on iNOS and COX-2 expression, and on NO and PGE2 generation. CONCLUSION: These data show that melittin and bee venom prevent LPS and SNP-induced NO and PGE2 production via JNK pathway dependent inactivation of NF-kappaB, and suggest that inactivation of JNK pathways may also contribute to the anti-inflammatory and anti-arthritis effects of melittin and bee venom. PMID- 18507871 TI - Safety of intramuscular influenza vaccine in patients receiving oral anticoagulation therapy: a single blinded multi-centre randomized controlled clinical trial. AB - BACKGROUND: Influenza vaccines are recommended for administration by the intramuscular route. However, many physicians use the subcutaneous route for patients receiving an oral anticoagulant because this route is thought to induce fewer hemorrhagic side effects. Our aim is to assess the safety of intramuscular administration of influenza vaccine in patients on oral anticoagulation therapy. METHODS: DESIGN: Randomised, controlled, single blinded, multi-centre clinical trial. SETTING: 4 primary care practices in Barcelona, Spain. PARTICIPANTS: 229 patients on oral anticoagulation therapy eligible for influenza vaccine during the 2003-2004 season. INTERVENTIONS: intramuscular administration of influenza vaccine in the experimental group (129 patients) compared to subcutaneous administration in the control group (100 patients). PRIMARY OUTCOME: change in the circumference of the arm at the site of injection at 24 hours. SECONDARY OUTCOMES: appearance of local reactions and pain at 24 hours and at 10 days; change in INR (International Normalized Ratio) at 24 hours and at 10 days. Analysis was by intention to treat using the 95% confidence intervals of the proportions or mean differences. RESULTS: Baseline variables in the two groups were similar. No major side effects or major haemorrhage during the follow-up period were reported. No significant differences were observed in the primary outcome between the two groups. The appearance of local adverse reactions was more frequent in the subcutaneous administration group (37,4% vs. 17,4%, 95% confidence interval of the difference 8,2% to 31,8%). CONCLUSION: This study shows that the intramuscular administration route of influenza vaccine in patients on anticoagulant therapy does not have more side effects than the subcutaneous administration route. REGISTRATION NUMBER: NCT00137579 at clinicaltrials.gov. PMID- 18507872 TI - Calling on a million minds for community annotation in WikiProteins. AB - WikiProteins enables community annotation in a Wiki-based system. Extracts of major data sources have been fused into an editable environment that links out to the original sources. Data from community edits create automatic copies of the original data. Semantic technology captures concepts co-occurring in one sentence and thus potential factual statements. In addition, indirect associations between concepts have been calculated. We call on a 'million minds' to annotate a 'million concepts' and to collect facts from the literature with the reward of collaborative knowledge discovery. The system is available for beta testing at http://www.wikiprofessional.org. PMID- 18507873 TI - Do mental health problems in childhood predict chronic physical conditions among males in early adulthood? Evidence from a community-based prospective study. AB - BACKGROUND: Previous studies have documented associations between mental and physical health problems in cross-sectional studies, yet little is known about these relationships over time or the specificity of these associations. The aim of the current study was to examine the relationship between mental health problems in childhood at age 8 years and physical disorders in adulthood at ages 18-23 years. METHOD: Multiple logistic regression analyses were used to examine the relationship between childhood mental health problems, reported by child, parent and teacher, and physical disorders diagnosed by a physician in early adulthood. RESULTS: Significant linkages emerged between childhood mental health problems and obesity, atopic eczema, epilepsy and asthma in early adulthood. Specifically, conduct problems in childhood were associated with a significantly increased likelihood of obesity and atopic eczema; emotional problems were associated with an increased likelihood of epilepsy and asthma; and depression symptoms at age 8 were associated with an increased risk of asthma in early adulthood. CONCLUSIONS: Our findings provide the first evidence of an association between mental health problems during childhood and increased risk of specific physical health problems, mainly asthma and obesity, during early adulthood, in a representative sample of males over time. These data suggest that behavioral and emotional problems in childhood may signal vulnerability to chronic physical health problems during early adulthood. PMID- 18507874 TI - Psychological treatment of social anxiety disorder: a meta-analysis. AB - BACKGROUND: Older meta-analyses of the effects of psychological treatments of social anxiety disorder have found that these treatments have moderate to large effects. However, these earlier meta-analyses also included non-randomized studies, and there are many featured studies in this area which were published after the recent meta-analysis. METHOD: We conducted a systematic literature search and identified 29 randomized studies examining the effects of psychological treatments, with a total of 1628 subjects. The quality of studies varied. For the analyses, we used the computer program comprehensive meta analysis (version 2.2.021; Biostat, Englewood, NJ, USA). RESULTS: The mean effect size on social anxiety measures (47 contrast groups) was 0.70, 0.80 on cognitive measures (26 contrast groups) and 0.70 both on depression (19 contrast groups) and general anxiety measures (16 contrast groups). We found some heterogeneity, so we conducted a series of subgroup analyses for different variables of the studies. Studies with waiting-list control groups had significantly larger effect sizes than studies with placebo and treatment-as-usual control groups. Studies aimed at subjects who met Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria for social anxiety disorder had smaller effect sizes than studies in which other inclusion criteria were used. CONCLUSIONS: This study once more makes it clear that psychological treatments of social anxiety disorder are effective in adults, but that they may be less effective in more severe disorders and in studies in which care-as-usual and placebo control groups are used. PMID- 18507875 TI - Mental disorders in young adulthood. AB - BACKGROUND: The effect of mental disorders may be particularly detrimental in early adulthood, and information on mental disorders and their correlates in this age group is important. METHOD: A questionnaire focusing on mental health was sent to a nationally representative two-stage cluster sample of 1863 Finns aged 19 to 34 years. Based on a mental health screen, all screen-positives and a random sample of screen-negatives were asked to participate in a mental health assessment, consisting of the Structured Clinical Interview for DSM-IV (SCID-I) interview and neuropsychological assessment. We also obtained case-notes from all lifetime mental health treatments. This paper presents prevalences, sociodemographic associations and treatment contacts for current and lifetime mental disorders. RESULTS: Forty percent of these young Finnish adults had at least one lifetime DSM-IV Axis I disorder, and 15% had a current disorder. The most common lifetime disorders were depressive disorders (17.7%) followed by substance abuse or dependence (14.2%) and anxiety disorders (12.6%). Of persons with any lifetime Axis I disorder, 59.2% had more than one disorder. Lower education and unemployment were strongly associated with current and lifetime disorders, particularly involving substance use. Although 58.3% of persons with a current Axis I disorder had received treatment at some point, only 24.2% had current treatment contact. However, 77.1% of persons with a current Axis I disorder who felt in need of treatment for mental health problems had current treatment contact. CONCLUSIONS: Mental disorders in young adulthood are common and often co-morbid, and they may be particularly harmful for education and employment in this age group. PMID- 18507876 TI - The heritability of hedonic capacity and perceived stress: a twin study evaluation of candidate depressive phenotypes. AB - BACKGROUND: Anhedonia and stress sensitivity have been identified as promising depressive phenotypes. Research suggests that stress-induced anhedonia is a possible mechanism underlying the association between stress and depression. The present proof-of-concept study assessed whether hedonic capacity and stress perception are heritable and whether their genetic and environmental contributions are shared. METHOD: Twenty monozygotic (MZ) and 15 dizygotic (DZ) twin pairs completed a probabilistic reward task that provides an objective behavioral measure of hedonic capacity (reward responsiveness) and completed several questionnaires including the Perceived Stress Scale (PSS). Bivariate Cholesky models were used to investigate whether covariation between (1) depressive symptoms and hedonic capacity, (2) depressive symptoms and perceived stress, and (3) perceived stress and hedonic capacity resulted from shared or residual genetic and environmental factors. RESULTS: Additive genetic (A) and individual-specific environment (E) factors contributed to 46% and 54% of the variance in hedonic capacity, respectively. For perceived stress, 44% and 56% of the variance was accounted for by A and E factors, respectively. The genetic correlation between depression and hedonic capacity was moderate (ra=0.29), whereas the correlation between depression and stress perception was large (ra=0.67). Genetic and environmental correlations between hedonic capacity and stress perception were large (ra=0.72 and re=-0.43). CONCLUSIONS: The present study provides initial feasibility for using a twin approach to investigate genetic contributions of a laboratory-based anhedonic phenotype. Although these preliminary findings indicate that hedonic capacity and perceived stress are heritable, with substantial shared additive genetic contributions, replications in larger samples will be needed. PMID- 18507877 TI - Suicide in recently discharged psychiatric patients: a case-control study. AB - BACKGROUND: Few controlled studies have specifically investigated aspects of mental health care in relation to suicide risk among recently discharged psychiatric patients. We aimed to identify risk factors, including variation in healthcare received, for suicide within 3 months of discharge. METHOD: We conducted a national population-based case-control study of 238 psychiatric patients dying by suicide within 3 months of hospital discharge, matched on date of discharge to 238 living controls. RESULTS: Forty-three per cent of suicides occurred within a month of discharge, 47% of whom died before their first follow up appointment. The first week and the first day after discharge were particular high-risk periods. Risk factors for suicide included a history of self-harm, a primary diagnosis of affective disorder, recent last contact with services and expressing clinical symptoms at last contact with staff. Suicide cases were more likely to have initiated their own discharge and to have missed their last appointment with services. Patients who were detained for compulsory treatment at last admission, or who were subject to enhanced levels of aftercare, were less likely to die by suicide. CONCLUSIONS: The weeks after discharge from psychiatric care represent a critical period for suicide risk. Measures that could reduce risk include intensive and early community follow-up. Assessment of risk should include established risk factors as well as current mental state and there should be clear follow-up procedures for those who have self-discharged. Recent detention under the Mental Health Act and current use of enhanced levels of aftercare may be protective. PMID- 18507878 TI - Supplementation with orange and blackcurrant juice, but not vitamin E, improves inflammatory markers in patients with peripheral arterial disease. AB - Inflammation and endothelial activation are associated with an increased risk of CVD and epidemiological evidence suggests an association between levels of markers of inflammation or endothelial activation and the intake of fruit. Also, vitamin E, a fat-soluble antioxidant, has anti-inflammatory properties. We performed a randomised 2 x 2 factorial, crossover trial to determine the effect of orange and blackcurrant juice (500 ml/d) and vitamin E (15 mg RRR-alpha tocopherol/d) supplementation on markers of inflammation and endothelial activation in forty-eight patients with peripheral arterial disease. Patients were randomly allocated to two dietary supplements from the four possible combinations of juice and vitamin E: juice+vitamin E; juice+placebo; reference beverage (sugar drink)+vitamin E; and reference beverage+placebo. The supplementations were given for 28 d, separated by a 4-week wash-out period. Analysis of main effects showed that juice decreased C-reactive protein (CRP) by 11% and fibrinogen by 3% while the reference drink increased CRP by 13% and fibrinogen by 2% (P<0.008 and P<0.002, respectively). No significant differences were measured for IL-6 and the endothelial activation markers von Willebrand factor, tissue-plasminogen activator and plasmin activator inhibitor-1. Vitamin E supplementation had no significant effects on the various markers. We observed no significant interaction between juice and vitamin E. In this study, orange and blackcurrant juice reduced markers of inflammation, but not markers of endothelial activation, in patients with peripheral arterial disease, relative to sugar drinks. PMID- 18507879 TI - Re: Accurate consent for insertion and later removal of grommets. PMID- 18507880 TI - Trans-10, cis-12-conjugated linoleic acid does not increase body fat loss induced by energy restriction. AB - Very little evidence exists concerning the effects of conjugated linoleic acid (CLA) on body fat reduction induced by energy restriction. Moreover, although an effect of trans-10, cis-12-CLA on lipolysis has been suggested, it has not been consistently shown. The aims of the present study were to determine whether trans 10, cis-12-CLA increases the reduction of body fat induced by energy restriction, and to analyse its effect on lipolysis and adipose tissue lipase expression (hormone-sensitive lipase (HSL) and adipose tissue TAG lipase (ATGL)). Male Syrian Golden hamsters were fed a high-fat diet during 7 weeks in order to make them fatter. Then they were submitted to a mild energy restriction (25 %) without or with supplementation of 0.5 % trans-10, cis-12-CLA for 3 weeks. Basal glycerol release and lipolysis stimulated by several drugs acting at different levels of the lipolytic cascade were measured in epididymal adipose tissue. The expression of HSL and ATGL was assessed by real-time RT-PCR. No differences were found in adipose tissues size between the experimental groups. Medium adipocyte size and total number of adipocytes were similar in both experimental groups. Animals fed the CLA-enriched diet showed similar lipolytic rates as well as HSL and ATGL expressions to the controls. In conclusion, trans-10, cis-12-CLA does not promote adipose tissue lipid mobilisation nor does it heighten body fat reduction induced by energy restriction. Consequently, this CLA isomer does not seem to be a useful tool to be included in body weight-loss strategies followed in obesity treatment. PMID- 18507881 TI - Cognitive and mood effects in healthy children during 12 weeks' supplementation with multi-vitamin/minerals. AB - Adequate levels of vitamins and minerals are essential for optimal neural functioning. A high proportion of individuals, including children, suffer from deficiencies in one or more vitamins or minerals. This study investigated whether daily supplementation with vitamins/minerals could modulate cognitive performance and mood in healthy children. In this randomised, double-blind, placebo controlled, parallel groups investigation, eighty-one healthy children aged from 8 to 14 years underwent laboratory assessments of their cognitive performance and mood pre-dose and at 1 and 3 h post-dose on the first and last days of 12 weeks' supplementation with a commercially available vitamins/mineral product (Pharmaton Kiddi). Interim assessments were also completed at home after 4 and 8 weeks at 3 h post-dose. Each assessment comprised completion of a cognitive battery, delivered over the Internet, which included tasks assessing mood and the speed and accuracy of attention and aspects of memory (secondary, semantic and spatial working memory). The vitamin/mineral group performed more accurately on two attention tasks: 'Arrows' choice reaction time task at 4 and 8 weeks; 'Arrow Flankers' choice reaction time task at 4, 8 and 12 weeks. A single task outcome (Picture Recognition errors) evinced significant decrements at 12 weeks. Mood was not modulated in any interpretable manner. Whilst it is possible that the significant improvements following treatment were due to non-significant numerical differences in performance at baseline, these results would seem to suggest that vitamin/mineral supplementation has the potential to improve brain function in healthy children. This proposition requires further investigation. PMID- 18507882 TI - Total plasma carotenoids and mortality in the elderly: results of the Epidemiology of Vascular Ageing (EVA) study. AB - Carotenoids are pigments found in fruits and vegetables. While high intakes of fruits and vegetables have been found to be associated with lower mortality, our objective is to investigate if total plasma carotenoids, via their antioxidant properties, are associated with mortality risk in a free-living elderly population. The 'Epidemiology of Vascular Ageing' (EVA) study (n 1389; 59-71 years) is a 9-year longitudinal study with six waves of follow-up. The association between baseline total plasma carotenoids and mortality was determined by Cox proportional hazards regression analyses. Low total plasma carotenoid level was significantly associated with all-cause mortality in men but not in women. After controlling for potential confounding factors, mortality risk increased significantly in men (P = 0.03) with plasma carotenoids in the lowest quintile compared with men with plasma carotenoids in the highest (relative risk 2.94 (95% CI 1.21, 7.17)). A significant association between mortality by cancer and low plasma carotenoid level variable was also found in men (unit = 1 micromol/l; relative risk 1.72 (95% CI 1.02, 2.86); P = 0.01). Associations between total plasma carotenoids and mortality risk remained statistically significant after taking into account: (1) plasma Se level, which previously was found associated with mortality in this population and (2) thiobarbituric acid reactive substances level considered as an indicator of oxidative stress. By showing, prospectively, in a general healthy elderly population, that total plasma carotenoid levels were independently associated with mortality risk in men, the present study suggests that total plasma carotenoid levels could be a health indicator in elderly populations. PMID- 18507883 TI - Twenty years' demographic change in sedentes and migrants of an international migrant-sending community in Tonga. AB - In the Kingdom of Tonga, migration to overseas developed countries has prevailed. To elucidate the effects of migration on population dynamics, an interview survey was conducted in the migrant-sending community of Kolovai, in the outer region of Tongatapu Island. All births, deaths, marriages and in- and out-migrations that took place between 1983 and 2002 were recorded for all members of the 'Kolovai population', consisting of persons who had lived in this community for at least a one-year period during this 20 years. The 'Kolovai population' members, numbering 1184 (564 males and 620 females), were divided into three groups based on residence at the end of each year, i.e. Kolovai (called KK), other places in Tonga (KT) or overseas countries (KO). The KK population decreased from 774 in 1982 to 570 in 2002, owing mostly to an increase of 167 persons as the natural balance and a decrease of 324 persons as the balance of international migration. Comparison of total fertility rate (TFR) between KK and KO women revealed that the mean TFR of the former decreased from 3.460 in the earlier 10-year period (1983-1992) to 2.240 in the later 10-year period (1993-2002), while that of the latter was more than 3.5 in both 10-year periods. This difference was largely due to the decrease in the proportion married among KK women. If the current trends of international migration and fertility continue, the population of Kolovai will be reduced and its age composition will become cylinder-shaped in the near future. PMID- 18507884 TI - Localization of carbohydrate determinants common to Biomphalaria glabrata as well as to sporocysts and miracidia of Schistosoma mansoni. AB - The presence of antigenic carbohydrate epitopes shared by Biomphalaria glabrata as well as by the sporocysts and miracidia representing snail-pathogenic larval stages of Schistosoma mansoni was assayed by immunohistochemical staining of paraformaldehyde-fixed tissues. To this end, both polyclonal rabbit antiserum raised against soluble egg antigens (SEA) of S. mansoni and monoclonal antibodies recognizing the carbohydrate epitopes LDN [GalNAc(beta1-4)GlcNAc(beta1-)], F-LDN [Fuc(alpha1-3)GalNAc(beta1-4)GlcNAc(beta1-)], LDN-F [GalNAc(beta1-4)[Fuc(alpha1 3)]GlcNAc(beta1-)], LDN-DF [GalNAc(beta1-4)[Fuc(alpha1-2)Fuc(alpha1 3)]GlcNAc(beta1-)] and Lewis X [Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-)] were used. Intriguingly, anti-SEA serum as well as anti-F-LDN antibodies displayed significant binding in the foot region, anterior tissue and the hepatopancreas of uninfected snails, whereas the Lewis X epitope was only weakly detectable in the latter tissue. In contrast, increased binding of antibodies recognizing LDN, LDN F and LDN-DF was observed in infected snail tissue, in particular in regions involved in sporocystogenesis, in addition to an enhanced binding of anti-SEA serum and antibodies reacting with F-LDN. A pronounced expression of most of these carbohydrate antigens was also observed at the surface of miracidia. Hence, the detection of shared carbohydrate determinants in uninfected snail tissue, sporocysts and miracidia may support the hypothesis of carbohydrate-based molecular mimicry as a survival strategy of S. mansoni. PMID- 18507885 TI - Failure to deactivate in the prefrontal cortex in schizophrenia: dysfunction of the default mode network? AB - BACKGROUND: Functional imaging studies using working memory tasks have documented both prefrontal cortex (PFC) hypo- and hyperactivation in schizophrenia. However, these studies have often failed to consider the potential role of task-related deactivation. METHOD: Thirty-two patients with chronic schizophrenia and 32 age- and sex-matched normal controls underwent functional magnetic resonance imaging (fMRI) scanning while performing baseline, 1-back and 2-back versions of the n back task. Linear models were used to obtain maps of activations and deactivations in the groups. RESULTS: The controls showed activation in the expected frontal regions. There were also clusters of deactivation, particularly in the anterior cingulate/ventromedial PFC and the posterior cingulate cortex/precuneus. Compared to the controls, the schizophrenic patients showed reduced activation in the right dorsolateral prefrontal cortex (DLPFC) and other frontal areas. There was also an area in the anterior cingulate/ventromedial PFC where the patients showed apparently greater activation than the controls. This represented a failure of deactivation in the schizophrenic patients. Failure to activate was a function of the patients' impaired performance on the n-back task, whereas the failure to deactivate was less performance dependent. CONCLUSIONS: Patients with schizophrenia show both failure to activate and failure to deactivate during performance of a working memory task. The area of failure of deactivation is in the anterior prefrontal/anterior cingulate cortex and corresponds to one of the two midline components of the 'default mode network' implicated in functions related to maintaining one's sense of self. PMID- 18507886 TI - Women's awareness of the importance of long-chain omega-3 polyunsaturated fatty acid consumption during pregnancy: knowledge of risks, benefits and information accessibility. AB - OBJECTIVE: To evaluate pregnant women's knowledge regarding the importance of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) consumption during pregnancy and assess their views on current information availability. DESIGN: A 27-item demographic and food safety/behaviour questionnaire was administered to pregnant women during their antenatal clinic visits. chi2 tests were performed using SPSS. SETTING: Antenatal clinics at two regional hospitals in New South Wales, Australia. SUBJECTS: One hundred and ninety (n 190) pregnant women. RESULTS: Three quarters of the women had not received information regarding LC n 3 PUFA. Approximately half of the women were aware of issues relating to LC n-3 PUFA; however, their knowledge was limited, with most obtaining their knowledge from books and magazines. Women generally had low (30 %, 29 %) to moderate (28 %, 24 %) levels of concern about LC n-3 PUFA and mercury, respectively. CONCLUSIONS: Pregnant women lack knowledge of LC n-3 PUFA and health-care services do not provide pregnant women with adequate information on the importance of eating foods high in LC n-3 PUFA during pregnancy. PMID- 18507887 TI - The predictive value of different socio-economic indicators for overweight in nine European countries. AB - OBJECTIVES: To assess which socio-economic indicator best predicts overweight in the European Union: educational attainment, occupational class or household income. SETTING: The prevalence of overweight is strongly related to socio economic position. The relative importance of different socio-economic dimensions is uncertain, and might vary between countries. DESIGN AND SUBJECTS: Cross sectional self-report data of the European Community Household Panel were obtained from nine countries (n 52,855; age 25-64 years). Uni- and multivariate regression analyses were employed to predict overweight (BMI >or= 25 kg/m2) in relationship to socio-economic indicators. Occupational class was measured using the new European Socioeconomic Classification. RESULTS: Large socio-economic differences in overweight were observed in all countries, especially for women. For both sexes, a low educational attainment was the strongest predictor of overweight. After controlling for education, overweight was negatively related to household income in women, but positively in men. Similar patterns were found for occupational class. For women, but not for men, educational inequalities in overweight were generally greater in Southern European countries. A similar pattern of inequalities in overweight was observed for all ages between 25 and 64 years. CONCLUSIONS: Across Europe, overweight was more strongly and more consistently related to educational attainment than to occupational class or household income. People with lower educational attainment should be a specific target group for programmes and policies that aim to prevent overweight. PMID- 18507888 TI - The Asian enigma: predisposition for low adult BMI among people of South Asian descent. AB - OBJECTIVE: To investigate the Asian enigma, the phenomenon of relatively high levels of undernutrition among children and adult women in South Asia, despite more favourable records with respect to infant mortality, women's education, food availability or other aspects of living conditions in comparison with, for example, sub-Saharan Africa. DESIGN: Literature has been explored to identify countries outside South Asia that are home to sizeable population segments from different ethnic backgrounds, including people of South Asian and African descent, and to compare prevalence rates of undernutrition in combination with indicators of standard of living between these various population segments. RESULTS: Data on adult undernutrition prevalence rates among population groups of different ethnic descent living in the same country (South Africa, Fiji and the USA) generally reveal the highest prevalence rates of low BMI among adults, males and females, from South Asian background. The relatively high rates of low BMI among adults from South Asian background cannot be explained by less favourable socio-economic characteristics, such as lower income or less access to food. CONCLUSION: It is hypothesized that there exists among adults of South Asian descent an ethnic predisposition for a low BMI. Other factors that may contribute to high levels of undernutrition in South Asia are discrimination of women and a poor dietary quality of poor households' food composition pattern. The question needs to be addressed whether the commonly used cut-off point for adult underweight (BMI < 18.5 kg/m2) is universally applicable or whether ethnic differences should be taken into account. PMID- 18507889 TI - Why do low-income women not use food stamps? Findings from the California Women's Health Survey. AB - OBJECTIVE: To determine factors associated with Food Stamp Program (FSP) participation in a potentially eligible California population. DESIGN: The California Women's Health Survey is an on-going annual telephone survey that collects data about health-related attitudes and behaviours from a randomly selected sample of women. Statistical procedures included chi2 and logistic regression. SETTING: California, USA, from 2002 to 2004. SUBJECTS: A total of 527 FSP female participants and 1405 potentially eligible non-participant females, aged 18 years and older. RESULTS: The following characteristics remained independently and positively related to FSP participation: single mother with children; unemployed; on welfare; on WIC (the Special Supplemental Nutrition Program for Women, Infants, and Children); and US-born. Women under 25 years and those over 54 years were less likely to participate than women aged 25-54 years. Hispanic/Latino ethnicity was also negatively related to participation. Over 42 % of potentially eligible non-participants cited 'don't need them' as the reason for not applying for FSP, but 34.9 % either do not think they are eligible or do not know how to apply. Potentially eligible non-participants who cite 'don't need them' as a reason for not applying are less likely to be food insecure, to have inadequate income and to use alternative emergency aid, compared to women citing all other reasons. The highest level of need is among those who cite worry about citizenship or stigma as reasons for not applying. CONCLUSION: Strategies to increase participation in FSP should incorporate messages that change the public's perception of the programme, in addition to simplifying the application process, raising awareness of eligibility criteria and improving customer service. PMID- 18507891 TI - Costs, quality of life, treatment satisfaction and compliance in patients with beta-thalassemia major undergoing iron chelation therapy: the ITHACA study. AB - OBJECTIVES: Iron chelation treatment (ICT) in beta-thalassemia major (beta-TM) patients undergoing blood transfusions can cause low satisfaction, low compliance, with possible negative consequences on treatment success, patients' wellbeing, and costs. The purpose was to estimate the societal burden attributable to beta-TM in terms of direct and indirect costs, health-related quality-of-life (HRQoL), satisfaction and compliance with ICT in patients undergoing transfusions and ICT. RESEARCH DESIGN AND METHODS: The naturalistic, multicenter, longitudinal Italian-THAlassemia-Cost-&-Outcomes-Assessment (ITHACA) cost-of-illness study was conducted involving patients of any age, on ICT for at least 3 years, who were enrolled at 8 Italian Thalassemia Care Centers. Costs were estimated from the societal perspective, quantified with tariffs, prices, or net earnings valid in 2006. RESULTS: One-hundred and thirty-seven patients were enrolled (median age = 28.3, 3-48 years, 49.6% male) and retrospectively observed for a median of 11.6 months. Mean direct costs were euro1242/patient/month, 55.5% attributable to ICT, 33.2% attributable to transfusions. Relevant quantity and quality of productivity was lost. Both physical and mental components of HRQoL were compromised. Little difficulties remembering to take ICT and positive satisfaction with the perceived effectiveness of therapy were declared, but not good levels of satisfaction with acceptance, perception of side effects and burden of ICT. CONCLUSIONS: The management of beta-TM patients undergoing transfusions and ICT is efficacious, although costly, but overall benefits were not always perceived as optimal by patients. Efforts must be focused to improve patients' acceptance and satisfaction with their therapy; this would contribute to a better compliance and hence an increase in treatment effectiveness and patients' overall wellbeing, with expected improved allocation of human and economic resources. PMID- 18507892 TI - Integrating evidence from multiple sources to evaluate post-approval safety: an example of sildenafil citrate and cardiovascular events. AB - OBJECTIVE: Recent high-profile medicine withdrawals have highlighted the complex decision-making process that regulators, pharmaceutical companies, prescribers, and patients must undertake in determining whether a drug has an appropriate benefit-risk balance. Our objective was to analyze the utility of different drug safety data sources and methods, using the experience of sildenafil citrate (Viagra) and post-approval concerns about its potential association with cardiovascular (CV) events (i.e., myocardial infarction [MI] and death) as a case study. METHODS: We evaluated safety data from three sources: the standard passive surveillance system (i.e., spontaneous reports filed to Pfizer Inc), pooled clinical trial data, and a prospective observational cohort study, the International Men's Health Study (IMHS). RESULTS: More than 28 000 spontaneous reports were received in the first 7 years after approval. Between 2001 and 2005, the proportion filed by persons other than healthcare professionals (61%) was approximately double the proportion averaged across five other drugs from the manufacturer's safety database. CV events and/or deaths represented 22.0% of reports, and 23% of reported deaths were medically unconfirmed reports made by persons other than healthcare professionals. In contrast, MI and all-cause mortality rates for sildenafil from both the pooled clinical trial data and the IMHS were similar to placebo, despite differences in methods and populations. CONCLUSIONS: These results suggest that passive surveillance may generate apparent signals of risk, as was the case with sildenafil and CV events. However, to adequately assess the benefit-risk profile of a drug, these signals must be evaluated via other data sources such as clinical trial and epidemiologic studies, as the apparent signal was not supported by more rigorously collected data. Our post-marketing analysis was unable to examine all potential influences of spontaneous reports, and the study data sources (although large for erectile dysfunction studies) were not designed to exclude small CV risks. PMID- 18507893 TI - Patient-physician interactions during early breast-cancer treatment: results from an international online survey. AB - OBJECTIVE: To examine, via an international survey, the impact of positive and negative interactions on the patient-physician relationship. RESEARCH DESIGN AND METHODS: This online survey was conducted in five countries (USA, UK, Germany, Italy and France) in two populations: (1) postmenopausal women diagnosed with early breast cancer (EBC) in the past 5 years, who had undergone surgery; (2) breast-cancer physicians. The survey covered several topics: patient-physician interactions, EBC treatment options and sources of information. RESULTS: In total, 462 physicians and 600 patients responded. Most (85%) physicians considered having a good relationship with their patient the most rewarding part of their job. Although 60% of physicians were satisfied with the consultation time (average 17.9 min), 30% considered it insufficient, whilst 49% of patients would prefer more time. Patients reported that physicians were a primary source of information, with 81% indicating that trust in their physician was a vital component of their care. Many physicians (63%) felt that patients are overwhelmed by the amount of information available, but only 16% of patients felt overwhelmed. Most physicians (78%) consider that telling a patient she has EBC is easier than talking about recurrence; 44% rated talking about recurrence as the most stressful part of their job. Most physicians (91%) considered availability of clinical trial data to be crucial for building trust and 74% believed that treatments that minimise recurrence enable more positive conversations. CONCLUSIONS: Good-quality patient care extends beyond effective treatment to include good communication about therapeutic options, side effects, and the development of trust and confidence. The survey revealed some disparities in physicians' and patients' views, but demonstrated that a strong patient-physician relationship is highly valued by both. Patients need access to accurate information and adequate consultation time. Providing effective and well tolerated treatments that minimise recurrence may help promote positive interactions. PMID- 18507894 TI - MRI total sagittal abdominal diameter as a predictor of metabolic syndrome compared to visceral fat at L4-L5 level. AB - OBJECTIVE: To compare associations between anteroposterior (AP) diameter or sagittal abdominal diameter - a measure of total central fat, and visceral fat alone with the metabolic syndrome as defined by ATPIII criteria. RESEARCH DESIGN AND METHODS: Twenty-four Caucasian male with type 2 diabetes and 24 non-diabetic Caucasian male subjects [body mass index (BMI) (+/-SD): 32.23 +/- 7.52 kg/m(2), age (+/-SD): 51.35 +/- 13.80 years] were studied by magnetic resonance imaging (MRI) scan to measure central fat at L4-L5 level. The visceral and total central adipose tissue was calculated in cm(2) and total sagittal MRI diameter and visceral sagittal MRI diameters in cm. Components of the ATPIII definition of the metabolic syndrome and circulating adipocytokine concentrations were also measured. RESULTS: MRI total sagittal abdominal diameter was positively associated with waist circumference in controls (r=0.62, p=0.007) and in diabetic subjects (r=0.81, p<0.001). Binary logistic regression analysis showed that MRI calculated total sagittal diameter (r=0.61, p=0.002) was a more significant predictor of the adverse metabolic profile of the metabolic syndrome than MRI assessed visceral fat. Receiver operating characteristic curves revealed that MRI calculated total sagittal diameter most effectively identified subjects with the metabolic syndrome. CONCLUSIONS: MRI-calculated total sagittal abdominal diameter is a non-validated MRI method that predicts the adverse metabolic profile of the ATPIII definition of the metabolic syndrome. Antero-posterior fat is a dimension of central fat that seems to be more closely associated with cardiovascular risk compared to visceral fat. PMID- 18507895 TI - Efficacy, safety, and tolerability of desvenlafaxine 50 mg/day and 100 mg/day in outpatients with major depressive disorder. AB - OBJECTIVE: To assess the efficacy, safety, and tolerability of 50- and 100-mg/day doses of desvenlafaxine (administered as desvenlafaxine succinate), a serotonin norepinephrine reuptake inhibitor, for the treatment of major depressive disorder (MDD). RESEARCH DESIGN AND METHODS: Patients with Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) MDD and 17-item Hamilton Rating Scale for Depression (HAM-D(17)) scores > or =20 were randomly assigned to double-blind placebo or desvenlafaxine treatment (fixed dose of 50 mg/day or 100 mg/day) for 8 weeks. The primary efficacy measure was the HAM-D(17). Changes from baseline in HAM-D(17) scores were analyzed using analysis of covariance. The final on-therapy evaluation was the primary endpoint for efficacy analyses, using last-observation-carried-forward data. MAIN OUTCOMES MEASURES AND RESULTS: The intent-to-treat population included 447 patients. Desvenlafaxine 50 mg was associated with a significantly greater adjusted mean change from baseline on the HAM-D(17) (-11.5) compared with placebo (-9.5, p=0.018); the 100-mg dose group ( 11.0) did not achieve statistical significance (p=0.065). The 100-mg dose group experienced significant improvements compared with placebo on several secondary efficacy measures, including the 6-item Hamilton Depression Rating Scale (p=0.038) and the Visual Analog Scale-Pain Intensity total score (p=0.041). Both desvenlafaxine doses were generally well-tolerated. The most common adverse events (incidence > or =10% in either desvenlafaxine group and twice the rate of placebo) were dry mouth, constipation, insomnia, decreased appetite, hyperhidrosis, and dizziness. CONCLUSIONS: These results demonstrate efficacy, safety, and tolerability of desvenlafaxine 50 mg/day for treating MDD. The significant findings on secondary measures support the efficacy of desvenlafaxine 100 mg, as seen in other trials. Conclusions may be limited by the exclusion of MDD patients with comorbid conditions and the short-term desvenlafaxine treatment duration. PMID- 18507896 TI - Major issues and challenges of influenza pandemic preparedness in developing countries. AB - Better preparedness for an influenza pandemic mitigates its impact. Many countries have started developing and implementing national influenza pandemic preparedness plans. However, the level of preparedness varies among countries. Developing countries encounter unique and difficult issues and challenges in preparing for a pandemic. Deaths attributable to an influenza pandemic could be substantially higher in developing countries than in industrialized countries. Pharmaceutical interventions such as vaccines and antiviral agents are less likely to be available in developing countries. The public health and clinical infrastructure of developing countries are often inadequate to deal with a widespread health crisis such as an influenza pandemic. Such an event will inevitably have a global effect. Therefore, improving pandemic preparedness in every country, particularly developing ones, is urgently needed. PMID- 18507897 TI - Managing potential laboratory exposure to ebola virus by using a patient biocontainment care unit. AB - In 2004, a scientist from the US Army Medical Research Institute of Infectious Diseases (USAMRIID) was potentially exposed to a mouse-adapted variant of the Zaire species of Ebola virus. The circumstances surrounding the case are presented, in addition to an update on historical admissions to the medical containment suite at USAMRIID. Research facilities contemplating work with pathogens requiring Biosafety Level 4 laboratory precautions should be mindful of the occupational health issues highlighted in this article. PMID- 18507898 TI - Transmission of human papillomavirus in heterosexual couples. AB - We examined the transmission of human papillomavirus (HPV) in 25 heterosexual, monogamous couples (25 men, 25 women), followed up over an average of 7.5 months. A total of 53 heterosexual transmission events were observed among 16 couples (14 male-to-female and 39 female-to male). Sexual transmission involved 13 different oncogenic and nononcogenic HPV types; 8% were vaccine-covered types transmitted between partners. The overall rate of HPV transmission from the penis to the cervix was 4.9/100 person-months, which was substantially lower than that from the cervix to the penis (17.4/100 person-months). Transmission between the hands and genitals, as well as apparent self-inoculation events (primarily in men), were also observed. Couples who transmitted HPV were more sexually active and used condoms less frequently. These results have implications for HPV prevention and control strategies, including the targeting of prophylactic vaccines. PMID- 18507899 TI - Population-attributable risk estimates for risk factors associated with Campylobacter infection, australia. AB - In 2001-2002, a multicenter, prospective case-control study involving 1,714 participants > or =5 years of age was conducted in Australia to identify risk factors for Campylobacter infection. Adjusted population-attributable risks (PARs) were derived for each independent risk factor contained within the final multivariable logistic regression model. Estimated PARs were combined with adjusted (for the > or =5 years of age eligibility criterion) notifiable disease surveillance data to estimate annual Australian Campylobacter case numbers attributable to each risk factor. Simulated distributions of "credible values" were then generated to model the uncertainty associated with each case number estimate. Among foodborne risk factors, an estimated 50,500 (95% credible interval 10,000-105,500) cases of Campylobacter infection in persons > or =5 years of age could be directly attributed each year to consumption of chicken in Australia. Our statistical technique could be applied more widely to other communicable diseases that are subject to routine surveillance. PMID- 18507900 TI - Influenza A virus (H3N8) in dogs with respiratory disease, Florida. AB - In 2004, canine influenza virus subtype H3N8 emerged in greyhounds in the United States. Subsequent serologic evidence indicated virus circulation in dog breeds other than greyhounds, but the virus had not been isolated from affected animals. In 2005, we conducted virologic investigation of 7 nongreyhound dogs that died from respiratory disease in Florida and isolated influenza subtype H3N8 virus. Antigenic and genetic analysis of A/canine/Jacksonville/2005 (H3N8) and A/canine/Miami/2005 (H3N8) found similarity to earlier isolates from greyhounds, which indicates that canine influenza viruses are not restricted to greyhounds. The hemagglutinin contained 5 conserved amino acid differences that distinguish canine from equine lineages. The antigenic homogeneity of the canine viruses suggests that measurable antigenic drift has not yet occurred. Continued surveillance and antigenic analyses should monitor possible emergence of antigenic variants of canine influenza virus. PMID- 18507901 TI - Tuberculosis from Mycobacterium bovis in binational communities, United States. AB - The epidemiology of tuberculosis (TB) in the United States is changing as the incidence of disease becomes more concentrated in foreign-born persons. Mycobacterium bovis appears to be contributing substantially to the TB incidence in some binational communities with ties to Mexico. We conducted a retrospective analysis of TB case surveillance data from the San Diego, California, region from 1994 through 2005 to estimate incidence trends, identify correlates of M. bovis disease, and evaluate risk factors for deaths during treatment. M. bovis accounted for 45% (62/138) of all culture-positive TB cases in children (<15 years of age) and 6% (203/3,153) of adult cases. M. bovis incidence increased significantly (p = 0.002) while M. tuberculosis incidence declined (p<0.001). Almost all M. bovis cases from 2001 through 2005 were in persons of Hispanic ethnicity. Persons with M. bovis were 2.55x (p = 0.01) as likely to die during treatment than those with M. tuberculosis. PMID- 18507902 TI - Validation of syndromic surveillance for respiratory pathogen activity. AB - Syndromic surveillance is increasingly used to signal unusual illness events. To validate data-source selection, we retrospectively investigated the extent to which 6 respiratory syndromes (based on different medical registries) reflected respiratory pathogen activity. These syndromes showed higher levels in winter, which corresponded with higher laboratory counts of Streptococcus pneumoniae, respiratory syncytial virus, and influenza virus. Multiple linear regression models indicated that most syndrome variations (up to 86%) can be explained by counts of respiratory pathogens. Absenteeism and pharmacy syndromes might reflect nonrespiratory conditions as well. We also observed systematic syndrome elevations in the fall, which were unexplained by pathogen counts but likely reflected rhinovirus activity. Earliest syndrome elevations were observed in absenteeism data, followed by hospital data (+1 week), pharmacy/general practitioner consultations (+2 weeks), and deaths/laboratory submissions (test requests) (+3 weeks). We conclude that these syndromes can be used for respiratory syndromic surveillance, since they reflect patterns in respiratory pathogen activity. PMID- 18507903 TI - Antibodies against Lagos bat virus in megachiroptera from West Africa. AB - To investigate the presence of Lagos bat virus (LBV)-specific antibodies in megachiroptera from West Africa, we conducted fluorescent antibody virus neutralization tests. Neutralizing antibodies were detected in Eidolon helvum (37%), Epomophorus gambianus (3%), and Epomops buettikoferi (33%, 2/6) from Ghana. These findings confirm the presence of LBV in West Africa. PMID- 18507904 TI - Increase in adult Clostridium difficile-related hospitalizations and case fatality rate, United States, 2000-2005. AB - Virulence of and deaths from Clostridium difficile-associated disease (CDAD) are on the rise in the United States. The incidence of adult CDAD hospitalizations doubled from 5.5 cases per 10,000 population in 2000 to 11.2 in 2005, and the CDAD-related age-adjusted case-fatality rate rose from 1.2% in 2000 to 2.2% in 2004. PMID- 18507905 TI - Syphilis and hepatitis B Co-infection among HIV-infected, sex-trafficked women and girls, Nepal. AB - Sex trafficking may play a major role in spread of HIV across South Asia. We investigated co-infection with HIV and other sexually transmitted diseases among 246 sex-trafficked women and girls from Nepal. Those who were HIV positive were more likely than those who were HIV negative to be infected with syphilis and/or hepatitis B. PMID- 18507906 TI - Underreporting of human alveolar echinococcosis, Germany. AB - We estimated the total number of human alveolar echinococcosis cases in Germany from 2003 through 2005 using the multiple source capture-recapture method. We found a 3-fold higher incidence of the disease than that shown by national surveillance data. We propose a revision of the reporting system to increase case ascertainment. PMID- 18507907 TI - Rickettsial Seroepidemiology among farm workers, Tianjin, People's Republic of China. AB - High seroprevalence rates for Anaplasma phagocytophilum (8.8%), Coxiella burnetii (6.4%), Bartonella henselae (9.6%), and Rickettsia typhi (4.1%) in 365 farm workers near Tianjin, People's Republic of China, suggest that human infections with these zoonotic bacteria are frequent and largely unrecognized. Demographic features of seropositive persons suggest distinct epidemiology, ecology, and risks. PMID- 18507908 TI - Persistence of Yersinia pestis in soil under natural conditions. AB - As part of a fatal human plague case investigation, we showed that the plague bacterium, Yersinia pestis, can survive for at least 24 days in contaminated soil under natural conditions. These results have implications for defining plague foci, persistence, transmission, and bioremediation after a natural or intentional exposure to Y. pestis. PMID- 18507910 TI - Global distribution of novel rhinovirus genotype. AB - Global surveillance for a novel rhinovirus genotype indicated its association with community outbreaks and pediatric respiratory disease in Africa, Asia, Australia, Europe, and North America. Molecular dating indicates that these viruses have been circulating for at least 250 years. PMID- 18507911 TI - Prevalence of Cryptosporidium spp. and Giardia intestinalis in swimming pools, Atlanta, Georgia. AB - Cryptosporidium spp. and Giardia intestinalis have been found in swimming pool filter backwash during outbreaks. To determine baseline prevalence, we sampled pools not associated with outbreaks and found that of 160 sampled pools, 13 (8.1%) were positive for 1 or both parasites; 10 (6.2%) for Giardia sp., 2 (1.2%) for Cryptosporidium spp., and 1 (0.6%) for both. PMID- 18507912 TI - Spatial and temporal evolution of bluetongue virus in wild ruminants, Spain. AB - We confirmed the emergence of bluetongue virus (BTV) in 5 wild ruminant species in Spain. BTV seroprevalence was high and dispersed with time, with a south-to north gradient. Our results suggest a complex epidemiology of BTV and underline the need for additional research on wildlife in Europe. PMID- 18507913 TI - Ceftriaxone-resistant salmonella enterica serotype Newport, France. AB - The multidrug-resistant (MDR) Salmonella enterica serotype Newport strain that produces CMY-2 beta-lactamase (Newport MDR-AmpC) was the source of sporadic cases and outbreaks in humans in France during 2000-2005. Because this strain was not detected in food animals, it was most likely introduced into France through imported food products. PMID- 18507914 TI - Avian influenza A virus (H5N1) outbreaks, Kuwait, 2007. AB - Phylogenetic analysis of influenza A viruses (H5N1) isolated from Kuwait in 2007 show that (H5N1) sublineage clade 2.2 viruses continue to spread across Europe, Africa, and the Middle East. Virus isolates were most closely related to isolates from central Asia and were likely vectored by migratory birds. PMID- 18507915 TI - Land use and west nile virus seroprevalence in wild mammals. AB - We examined West Nile virus (WNV) seroprevalence in wild mammals along a forest to-urban gradient in the US mid-Atlantic region. WNV antibody prevalence increased with age, urbanization, and date of capture for juveniles and varied significantly between species. These findings suggest several requirements for using mammals as indicators of transmission. PMID- 18507916 TI - High failure rates of melarsoprol for sleeping sickness, Democratic Republic of Congo. AB - A retrospective chart review of 4,925 human African trypanosomiasis patients treated with melarsoprol in 2001-2003 in Equateur Nord Province of the Democratic Republic of Congo showed a treatment failure rate of 19.5%. This rate increased over the 3 years. Relapse rates were highest in the central part of the province. PMID- 18507917 TI - Nosocomial Outbreaks Caused by Leuconostoc mesenteroides subsp. mesenteroides. AB - From July 2003 through October 2004, 42 patients became infected by strains of Leuconostoc mesenteroides subsp. mesenteroides (genotype 1) in different departments of Juan Canalejo Hospital in northwest Spain. During 2006, 6 inpatients, also in different departments of the hospital, became infected (genotypes 2-4). Parenteral nutrition was the likely source. PMID- 18507918 TI - Anaplasma phagocytophilum infection in Ixodes ricinus, Bavaria, Germany. AB - Anaplasma phagocytophilum DNA was detected by real-time PCR, which targeted the msp2 gene, in 2.9% of questing Ixodes ricinus ticks (adults and nymphs; n = 2,862), collected systematically from selected locations in Bavaria, Germany, in 2006. Prevalence was significantly higher in urban public parks in Munich than in natural forests. PMID- 18507919 TI - Internet- versus telephone-based local outbreak investigations. AB - We compared 5 locally conducted, Internet-based outbreak investigations with 5 telephone-based investigations. Internet-based surveys required less completion time, and response rates were similar for both investigation methods. Participant satisfaction with Internet-based surveys was high. PMID- 18507920 TI - Lack of serologic evidence of Neospora caninum in humans, England. AB - Retrospective testing of 3,232 serum samples from the general population and 518 serum samples from a high-risk group showed no evidence of human exposure to Neospora caninum in England. Results were obtained by using immunofluorescence antibody testing and ELISA to analyze frequency distribution. PMID- 18507922 TI - In memoriam: Joshua Lederberg (1925-2008). PMID- 18507923 TI - Lethal Mycobacterium massiliense sepsis, Italy. PMID- 18507924 TI - Bovine kobuviruses from cattle with diarrhea. PMID- 18507925 TI - Vibrio cholerae O1 hybrid El Tor strains, Asia and Africa. PMID- 18507926 TI - Mycobacterium avium subsp. hominissuis infection in 2 pet dogs, Germany. PMID- 18507927 TI - Serogroup Y meningococcal disease, Colombia. PMID- 18507928 TI - Inquilinus limosus and cystic fibrosis. PMID- 18507929 TI - Splenic rupture and malignant Mediterranean spotted fever. PMID- 18507930 TI - Acetobacter indonesiensis pneumonia after lung transplant. PMID- 18507931 TI - Coronavirus antibodies in bat biologists. PMID- 18507932 TI - Chagas disease in ancient hunter-gatherer population, Brazil. PMID- 18507933 TI - Coxiella burnetii in wild-caught filth flies. PMID- 18507934 TI - Conflict and emerging infectious diseases. PMID- 18507939 TI - "As the dew is dried up by the morning sun, so are mankind's sins at the sight of Himalaya". PMID- 18507940 TI - Comparison of road traffic fatalities and injuries in Iran with other countries. AB - OBJECTIVE: To compare fatalities and injuries in road traffic crashes (RTC) in Iran with other countries. METHODS: Data were obtained from national health sources of Iran. These data included population number, registered motor-vehicles number, number of RTCs and consequent fatalities and injuries from 1997 to 2006. RESULTS: RTC fatality and injury rates increased from 1997 to 2005, but decreased in 2006. The overall men/women ratio in the RTC fatalities was 4.2:1. High RTC fatality rate of 39 per 100 000 population in Iran was almost the same as some other developing countries. In Iran, RTC fatalities in recent years were almost twice as much as the highest rate among the European countries. CONCLUSIONS: This investigation shows that in spite of reduction of RTC fatality in Iran in 2006, it is still one of the highest in the world. Moreover, this paper describes the state of RTC-related parameters in a developing country in comparison with the developed countries. PMID- 18507941 TI - Radiographic predictors of residual low back pain after laminectomy for lumbar canal stenosis: a minimum of 6-year follow-up. AB - OBJECTIVE: To identify radiographic predictors of residual low back pain (LBP) after laminectomy for lumbar canal stenosis (LCS). METHODS: Clinical results and radiographic findings in 69 patients who underwent single level laminectomy for LCS were retrospectively reviewed. Patients who had an improvement in LBP scores evaluated by Japanese Orthopaedic Association (JOA) scoring system during the follow-up periods were classified as the recovery group, and others were classified as the non-recovery group. Patients'clinical data and radiographic parameters like lordosis angle, range of motion and intervertebral rotational angle were analyzed using binary logistic regression analysis to detect factors significantly related with the occurrence of residual LBP. RESULTS: The average preoperative JOA score of 14.8+/-5.05 improved to 21.59+/-5.51 at the final follow-up. Binary logistic regression analysis revealed that significant predictors of residual LBP were preoperative lumbar lordosis angle and range of motion. CONCLUSIONS: Our results suggest that patients with flat back and limited lumbar mobility before surgery tend to have poor results in terms of LBP. Therefore, these sagittal radiographic parameters should be taken into account when choosing laminectomy as the surgical option for LCS. PMID- 18507942 TI - Immunological study on the transplantation of an improved deproteinized heterogeneous bone scaffold material in tissue engineering. AB - OBJECTIVE: To observe the immune response after the transplantation of a deproteinized heterogeneous bone scaffold and provides the theoretic reference for clinical practice. METHODS: The fresh pig bone and deproteinized bone were transplanted respectively to establish BABL/C thigh muscle pouches model of male mice and take the samples for detection at 1, 2, 4, 6 weeks after operation. Lymphocyte stimulation index, subset analysis, serum specific antibody IgG, cytokine detection and topographic histologic reaction after implantation were investigated. RESULTS: After the transplantation of deproteinized bone, lymphocyte stimulation index, CD(4)(+) and CD(8)(+) T-lymphocyte subsets, serum specific antibody IgG and cytokines in deproteinized bone group were significantly lower than those in fresh pig bone group at each time point (P<0.05). The histological examination found that in fresh bone group at each time point, a large quantity of inflammatory cells infiltrated in the surrounding of bone graft, and they were mainly lymphocytes, including macrophages and monocytes. In deproteinized bone group, there were few inflammatory cells infiltration around bone graft one week after operation. The lymphocytes were decreased as time went by. At 6 weeks, fibroblasts and fibrous tissue grew into the graft, and osteoclasts and osteoprogenitor cells appeared on the verge. CONCLUSIONS: The established heterogeneous deproteinized bone has low immunogenicity and is a potentially ideal scaffold material for bone tissue engineering. PMID- 18507943 TI - Clinical study of a new approach to thoracolumbar surgery. AB - OBJECTIVE: The conventional approaches for treatment of thoracolumbar diseases require extensive surgical exposure, often leading to postoperative pain and morbidity. Thoracoscopic-assisted surgery in these regions usually requires an extended recovery period due to the placement of drainage. We developed an innovative retroperitoneal-extrapleural approach to thoracolumbar involvement by an extra-diaphragmatic technique using dedicated instruments. Neither incision nor reconstruction of the diaphragm was necessary. Exposure to the lateral part of the thoracolumbar vertebrae could be achieved without crus resection. This study is aimed to evaluate the clinical outcomes of this new surgical procedure. METHODS: A total of 9 cases (5 cases of thoracolumbar fracture-dislocation, 1 each of spinal infection, tumor, thoracolumbar scoliosis and ossification of posterior longitudinal ligament) were subjected to the study. The average age of the patients was 52.3 years. The results were compared with the control group consisting of thoracoscopic surgery subgroup (5 patients, mean age 52.1 years) and conventional surgery subgroup (12 patients, mean age 61.3 years). RESULTS: Compared with the control group, the average period of bed confinement and mean intra- and postoperative blood loss decreased significantly. Pulmonary complications were avoided in all cases. The surgical time was shortened, postoperative pain was reduced, and early postoperative ambulation became possible. CONCLUSION: The diaphragm-preserving retroperitoneal-extrapleural approach that we developed is a valid minimally invasive alternative for the treatment of thoracolumbar diseases. PMID- 18507944 TI - Properties of deproteinized bone for reparation of big segmental defect in long bone. AB - OBJECTIVE: To explore suitable scaffold material for big segmental long bone defect by studying the properties of the prepared deproteinized bone. METHODS: Cancellated bone were made as 30 mm x mm x 3 mm bone blocks from inferior extremity of pig femur along bone trabecula. The deproteinized bone was prepared with an improved method. Their morphological features, components, cell compatibility, mechanical and immunological properties were investigated respectively. RESULTS: Deproteinized bone maintained natural reticular pore system. The main organic material is collagen I and inorganic composition is hydroxyapatite. It has good mechanical properties, cell adhesion rate and histocompatibility. CONCLUSION: This deproteinized bone can be applicable as scaffold for reparation of big segmental defect in long bone. PMID- 18507945 TI - Application of damage control orthopedics in 41 patients with severe multiple injuries. AB - OBJECTIVE: To probe the feasibility and efficacy of damage control orthopedics (DCO) in treating severe multiple injuries. METHODS: A retrospective analysis was made on the clinical data of 41 patients (31 males and 10 females, aged 18-71 years, mean: 36.4) with multiple injuries admitted to our department and treated by DCO from January 1995 to December 2005. RESULTS: As a first-stage therapy, devascularization of internal iliac arteries was performed in 29 patients with pelvic fractures combined with massive bleeding, including ligation of bilateral internal iliac arteries in 21 patients and embolization of bilateral internal iliac arteries in 8. And early external fixation of pelvis was performed in 10 patients. Ten patients with severe multiple injuries combined with femoral fractures were managed with primary debridement and temporal external fixation and 2 patients with spinal fractures combined with spinal cord compression received simple laminectomy. Thirty-one patients received definite internal fixation after resuscitation in intensive care unit. The overall mortality rate was 12.1% (5/41) with an average injury severity score of 41.4. The main causes of death were hemorrhagic shock and associated injuries. Complications occurred in 7 patients including acute respiratory distress syndrome in 3 cases, thrombosis of right common iliac artery in 1, subphernic abscess in 2 and infection of deep wound in lower extremity in 1. After treatment, all the patients got cured. CONCLUSIONS: Prompt diagnosis and integrated treatment are keys to higher survival rate in patients with severe multiple injuries. In this condition, DCO is an effective and safe option. PMID- 18507946 TI - Vascular endothelial growth factor and its receptor expression during the process of fracture healing. AB - OBJECTIVE: To study the expression regularity of vascular endothelial growth factor (VEGF) during the process of fracture healing, and the type of VEGF receptor expressed in the vascular endothelial cells of the fracture site. METHODS: The fracture model was made in the middle part of left radius in 35 rabbits. The specimens from the fracture site were harvested at 8, 24, 72 hours and 1, 3, 5, 8 weeks, and then fixed, decalcified, and sectioned frozenly to detect the expression of VEGF and its receptor at the fracture site by in situ hybridization and immunochemical assays. RESULTS: VEGF mRNA and VEGF expression was detected in many kinds of cells at the fracture site during 8 hours to 8 weeks after fracture. Flt1 receptor of VEGF was found in the vascular endothelial cells at the fracture site during 8 hours to 8 weeks after fracture, and strong expression of flk1 receptor was detected from 3 days to 3 weeks after fracture. CONCLUSIONS: The expression of VEGF and flt1 receptor appears during the whole course of fracture healing, especially from 1 to 3 weeks. Flk1 receptor is highly expressed in a definite period after fracture. VEGF is proved to be involved in the vascular reconstruction and fracture healing. PMID- 18507947 TI - Treatment of osteonecrosis of femoral head with BMSCs-seeded bio-derived bone materials combined with rhBMP-2 in rabbits. AB - OBJECTIVE: To evaluate the effect of autologous bone marrow mesenchymal stem cells (BMSCs) seeded bio-derived bone materials (BBM) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in repairing defect of osteonecrosis of femoral head (ONFH). METHODS: Early-stage osteonecrosis in the left hip was induced in 36 adult New Zealand white rabbits (provided by the Animal Center of Guangxi Medical University, Nanning, China) after core decompression and delivery of liquid nitrogen into the femoral head. Then the animals were divided into three groups according to the type of implants for bone repair: 12 rabbits with nothing (Group I, the blank control group), 12 with BBM combined with rhBMP-2 (Group II), and 12 with BMSCs-seeded BBM combined with rhBMP-2 (Group III). At 4, 8, and 12 weeks after surgery, X-ray of the femoral head of every 4 rabbits in each group was taken, and then they were killed and the femoral heads were collected at each time point, respectively. Gross observation was made on the femoral heads. After hematoxylin and eosin staining, Lane-sandhu scores of X-ray and bone densitometry were calculated and the histomorphometric measurements were made for the new bone trabeculae. RESULTS: At 12 weeks after surgery, two femoral heads collapsed in Group I, but none in Group II or Group III. X-ray examination showed that the femoral heads in Group I had defect shadow or collapsed while those in Group II had a low density and those in Group III presented with a normal density. Histologically, the defects of femoral heads were primarily filled with no new bone but fibrous tissues in Group I. In contrast, new bone regeneration and fibrous tissues occurred in Group II and only new bone regeneration occurrd in Group III. Lane-sandhu scores of X-ray, bone mineral density and rate of new bone in trabecular area in Group III were higher significantly than those of the other two groups. CONCLUSIONS: Our findings indicate a superior choice of repairing the experimental defect of ONFH with BMSCs- seeded BBM combined with rhBMP-2. PMID- 18507948 TI - Treatment of ipsilateral femoral neck and shaft fractures. AB - OBJECTIVE: To investigate the clinical characteristics, treatment options and causes of misdiagnosis of ipsilateral femoral neck and shaft fractures. METHODS: Among 20 patients with ipsilateral femoral neck and shaft fractures, 19 were treated operatively and 1 was treated conservatively. Sixteen cases of femoral shaft fractures were treated by open reduction and internal fixation with compressive plate, and 2 cases were treated with interlocking intramedullary nailing. Eighteen femoral neck fractures were treated with cannulated screws. Another patient was treated with proximal femoral nail to fix both the neck and shaft. Delayed diagnosis for femoral neck fractures occurred in 2 cases preoperatively. RESULTS: A total of 19 patients were followed up. The follow up period ranged from 5 to 48 months with an average of 15 months. All the fractures were healed. CONCLUSION: For case of femoral shaft fracture caused by high energy injury, an AP pelvic film should be routinely taken. Once the femoral neck fracture is recognized, operative reduction and fixation should be performed in time. Femoral neck and shaft fractures should be fixed separately. PMID- 18507949 TI - An experimental study of nerve bypass graft. AB - OBJECTIVE: To study the use of a nerve ''bypass'' graft as a possible alternative to neurolysis or segmental resection with interposition grafting in the treatment of neuroma-in-continuity. METHODS: A sciatic nerve crush injury model was established in the Sprague-Dawley rat by compression with a straight hemostatic forceps. Epineurial windows were created proximal and distal to the injury site. An 8-mm segment of radial nerve was harvested and coaptated to the sciatic nerve at the epineurial window sites proximal and distal to the compressed segment (bypass group). A sciatic nerve crush injury without bypass served as a control. Nerve conduction studies were performed over an 8-week period. Sciatic nerves were then harvested and studied under transmission electron microscopy. Myelinated axon counts were obtained. RESULTS: Nerve conduction velocity was significantly faster in the bypass group than in the control group at 8 weeks (63.57 m/s+/-5.83 m/s vs. 54.88 m/s+/-4.79 m/s, P<0.01). Myelinated axon counts in distal segments were found more in the experimental sciatic nerve than in the control sciatic nerve. Significant axonal growth was noted in the bypass nerve segment itself. CONCLUSION: Nerve bypass may serve to augment peripheral axonal growth while avoiding further loss of the native nerve. PMID- 18507950 TI - Implanting hydroxyapatite-coated porous titanium with bone morphogenetic protein 2 and hyaluronic acid into distal femoral metaphysis of rabbits. AB - OBJECTIVE: To assess the osseointegration capability of hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 (BMP-2) and hyaluronic acid to repair defects in the distal femur metaphysis in rabbits. METHODS: Porous titanium implants were made by sintering titanium powder at high temperature, which were coated with hydroxyapatite by alkali and heat treatment and with BMP-2 combined with bone regeneration materials. And hyaluronic acid was further used as delivery system to prolong the effect of BMP-2. The implants were inserted into the metaphysis of the distal femur of rabbits. The animals were killed at 6, 12 and 24 weeks to accomplish histological and biomechanical analyses. RESULTS: According to the result of histological analysis, the osseointegration in BMP-2 group was better than that of the HA-coated porous titanium group. In push-out test, all the samples had bigger shear stress as time passed by. There was statistical difference between the two groups in 6 and 12 weeks but not in 24 weeks. CONCLUSION: Hydroxyapatite-coated porous titanium with BMP-2 and hyaluronic acid has a good effect in repairing defects of distal femur in rabbits, which is a fine biotechnology for future clinical application. PMID- 18507951 TI - Evaluation of multislice computed tomographic perfusion imaging and computed tomographic angiography on traumatic cerebral infarction. AB - OBJECTIVE: To evaluate the application value of multislice computed tomographic perfusion imaging (MSCTPI) and multislice computed tomographic angiography (MSCTA) on traumatic cerebral infarction. METHODS: MSCTA was performed on 10 patients who were initially diagnosed as traumatic cerebral infarction by normal conventional computed tomography (NCCT), among whom, 3 patients were examined by MSCTPI simultaneously. Reconstructed images of the intracranial artery were made with techniques of maximum intensity projection (MIP) and volume rendering (VR) from MSCTA scanning data. Then the graph of function of four parameters, regional cerebral blood flow (rCBF), regional cerebral blood volume (rCBV), mean transit time (MTT), and time to peak (TTP), acquired by the perfusing analysis software was obtained. RESULTS: Among the 10 patients with traumatic cerebral infarction, 6 showed complex type on NCCT, which depicted abnormality on MSCTA, and 4 showed simple type on NCCT, which had negative results on MSCTA. Among the 4 patients with abnormal great vessels, 2 suffered from stenosis or occlusion of the middle cerebral artery, 1 from spasm of the anterior cerebral artery, and 1 from spasm of the vertebral-basal artery. The image of MSCTPI of 1 patient with massive cerebral infarction on the right cerebral hemisphere confirmed by CT was smaller than those of the other patients, which showed occlusion of the ipsilateral middle cerebral artery on MSCTA. Among the 6 patients whose MSCTA showed no abnormality, 4 showed simple infarction and 2 showed complex infarction. The infarction focus of 5 patients occurred in the basal ganglia and 1 in the splenium of corpus callosum. Among the 2 cases of small cerebral infarction volume on NCCT, one was normal, the other showed hypoperfusion on MSCTPI and was normal on MSCTA. CONCLUSION: The combination of MSCTPI and MSCTA is very useful for evaluating the change of intracranial artery in ischemic regions and assessing the cerebral hemodynamic information of traumatic cerebral infarction. PMID- 18507952 TI - Coverage of soft tissue defect in palm with prefabricated flap. PMID- 18507953 TI - [Advantages of neoadjuvant therapy in rectal neoplasm]. PMID- 18507954 TI - [Neuroendocrine carcinoma of the breast. Our experience and a proposal of a therapeutic algorithm for a rare tumor]. AB - The neuroendocrine carcinoma of the breast is a very rare tumor. In this paper we describe our experience in 10 cases of neuroendocrine carcinoma of the breast, and an integrated diagnostic-therapeutic proposal for this tumor. Since no positive association has been shown between neuroendocrine differentiation and tumor size, staging, grading, survival and therefore prognosis, we consider that surgical therapy for neuroendocrine tumors of the breast should be the same as that performed in common invasive histotypes. Due to the presence of specific cellular receptors in neuroendocrine tumors of the breast, somatostatin has been claimed as a useful tool both for diagnostic (Octreoscan) and therapy (for metastatic disease). As for therapy, synthetic analogs show advantages versus native somatostatin, because of a longer half-life, and data from literature report encouraging results obtained by using radiolabelled somatostatin analogs. One of these is 90 Y-Dotatoc; we have already used it in patients with neuroendocrine tumors of the lung. Our algorithm for neuroendocrine tumors of the breast includes diagnostic scintigraphy with Octreoscan and receptor-mediated radiolabelled therapy with 90 Y-Dotatoc in patients with confirmed scintigraphic expression of somatostatin receptors in tumoral tissue. PMID- 18507955 TI - [Primary duodenal adenocarcinoma: report of three cases, prognostic factors and therapeutic approach]. AB - Three cases of histologically proven primary non-ampullary adenocarcinoma of the duodenum, observed in our Department from 2001 to 2004, are described. The cases were treated by pancreaticoduodenectomy, duodenal resection and transduodenal excision, respectively. The rarity of this pathology is documented by few retrospective studies and justifies discussion about the main prognostic factors and the best therapeutic approach. We analyze diagnostic, therapeutic and prognostic factors after a revision of literature. PMID- 18507956 TI - [New therapeutic strategies for the treatment of difficult wounds]. AB - BACKGROUND: The medical-surgical treatment of the difficult wounds represents a socio-sanitary problem in continuous growth, currently involving in our Country around 2,000,000 people. The "difficult wound" is a loss of cutaneous substances, usually due to multifactorial pathogenesis, that do not spontaneously lead to a complete recovery. Numerous studies in the literature have evidenced that the use of the advanced wound dressings allows to reach the best clinical and economic results in the process of recovery of the difficult wounds. The advanced would dressing assures a longer period of permanence on the injury and shorten the time of treatment and, as a consequence, it is required a smaller number of applications in comparison with the traditional medications. The Wound Bed Preparation (WBP) can be defined as the global and coordinate management of the cutaneous injury, enabling to chip off the local barriers to the recovery, or promoting the effectiveness of the innovative therapeutic instruments. The term advanced wound dressing indicates the dressing material having biocompatibility characteristics. The purpose of the advanced wound dressings is the one to create the ideal environment for the cicatrization process and isolate the wound from traumas and external infections. PATIENTS AND METHODS: The "Difficult Wounds" Unit of the Department of Plastic and Reconstructive Surgery of the Policlinico Umberto I in Rome, from January to December 2006, treated 570 patients (308 men and 262 women), whose age was between 2 days and 85 years, affected by ulcers of various nature. Among our cases, 200 patients were selected and randomly separated in two different groups: group A consisting of 100 patients entirely treated with traditional medications; group B composed by 100 patients treated with advanced dressings. Every patient has locally been treated with periodic and specific medications, according to the type of difficult wound, and subsequently they proceeded to find out how to treat the systemic factors causing ulcer. The patients underwent 3 times a week to medications in those cases presenting infection signs and 2 times a week in those cases where no infection signs were shown, for period varying from 1 month up to one year for the chronic forms. RESULTS: The results showed a higher percentage of recovery reached by using the advanced dressings. Group A showed the followings results: the 53% of patients recovered from wounds; the remaining 47% patients did'nt not recover but in 17% cases medications showed to be of some help in the preparation of the vascular bed for the execution of a definitive operation (application of grafts or local edges), while the remaining 30% has shown a scarce improvement of the injury and they are still under treatment. Group B showed the 65% of patients recovered from wounds; as for the remaining 35% not recovered patients, medications represented an auxiliary aid to the preparation of the vascular bed for the execution of a definitive operation (application of grafts or local edges) for the 15% of patients, while the remaining 20%, even if not completely recovered, showed a notable improvement of the injury (reduction of the dimensions and disappearance of the infection and improvement of the patient quality of life). CONCLUSIONS: In synthesis, it emerges that the advanced dressings, if correctly used, offer advantages in terms of clinical effectiveness (rapid recovery from the injury), patient quality of the life and cheapness. It has also to be considered that the difficult wound is often the epiphenomenon of a systemic illness. The difficult wound requires, therefore, a multidisciplinary treatment. PMID- 18507957 TI - [Integrated treatment of breast cancer: review of our experience]. AB - In the industrialized West countries the breast cancer represents the most frequent malignant neoplasia in the women. From over 30 years we have a progressive increase of frequency of breast carcinoma. Mortality is substantially unchanged. The Authors, on the base of casuistry accumulated in five years of activity, trace a diagnostic-therapeutic run to the purpose to favour a line of behavior, practice and actual, for the management of breast cancer. PMID- 18507958 TI - Primary breast lymphoma: case reports and review of the literature. AB - As the prevalence of breast cancer has increased over recent years many cases with unusual presentation are emerging. This review attempts to identify different types of primary malignancies that can affect the breast. A five years review was undertaken at the Breast Unit of Azienda Ospedaliera "Santa Maria", Terni (Italy). All cases of breast malignancy admitted to the Breast Unit between 2001 and 2005 were reviewed: 750 patients were diagnosed with malignant breast disease. Primary breast lymphoma (was found only in two cases accounting for an incidence of 0,26%. The pre-operative diagnosis was infiltrating carcinoma in both cases and the patients were treated with surgery and systemic chemotherapy; currently they are disease- free. PMID- 18507959 TI - Intracerebral haemorrhage following surgical evacuation of chronic subdural haematoma: case report. AB - Intracerebral haemorrhages occurring after removal of chronic subdural haematoma have been reported as a rare but nearly uniformly devastating postoperative occurrence with incidence ranges between 0,7 and 5%. The absence of a particular attention paid to such complication in literature prompted us to present this report adding our personal experience to the cases already described in the literature. PMID- 18507960 TI - [A case of lymphoepithelial cyst (branchial cyst) in an elderly patient: diagnosis differential, treatment and literature review]. AB - We present a case of lateral cervical cyst stressing the difficulties about the diagnosis. The international guidelines for the management of lateral neck cysts in the over 40s' age group are taken in consideration. Our case is a 74 years old male patient with a 6 month history of a cervical swelling. We consider this case rare for the age of patient and the absence of malignancy. PMID- 18507961 TI - [Retroperitoneal giant mixed sarcoma. Case report]. AB - The Authors present a rare case of voluminous retroperitoneal tumor in a 41 years old woman. CT scan showed a solid abdominal expansive mass, with compression and displacement of the left lobe of the liver, spleen, stomach, pancreas, and left kidney. The patient underwent surgery for excision of a giant retroperitoneal mass. Surgery was uneventful. The patients recovered well. Histology showed a mixed liposarcoma and angiosarcoma with high grade of malignancy and positivity for vimentin, factor VIII, CD34, CD31 and negativity for S-100, CD68, AMS, AML. The prognosis of these tumours is closely related to local recurrence, histological type, size and radical surgery. A low-grade malignancy tumor, small sized and completely resected leads to a good prognosis. Radiation therapy and chemotherapy do not seem to have a strong influence on the prognosis. An aggressive surgical approach is the first choice for the treatment of such tumors. The resection of adjacent organs may be required for radical surgery. PMID- 18507962 TI - Usefullness of the ultrasonically activated scalpel in laparoscopic cholecystectomy: our experience and review of literature. AB - Laparoscopic cholecystectomy (LC) actually represents the most used and proper treatment for gallbladder lithiasis, because its many and known advantages in comparison with 'open' abdominal surgery. But there are some problems during and after LC due to the use of the electric scalpel and these have brought to the search of an alternative system of dissection and coagulation. The ultrasonically activated scalpel (Harmonic Scalpel, HS) allows to perform dissection and coagulation with a minimal thermal side effect for surrounding tissues, unlike the electrocoagulation. Furthermore, the use of the HS brings a series of advantages in comparison to the other electromagnetic forms of energy (electro scalpel, laser). HS cuts and coagulates with the same effectiveness of the electro-scalpel but, unlike this, it doesn't introduce risks of wandering currents. Moreover, HS contributes to have a more clean and clear (smokes-free) field of operation and it reduces the operative time, the bleeding and the costs of the operation without an increase of the complications and of the percentages of 'open' conversion, and perhaps leads to a less negative influence on the postoperative systemic immune response. The Authors report their experience that confirm these observations, according also with results reported in a brief review of the recent scientific literature, and support wider diffusion and technical development of this ultrasonically-operating surgical team. PMID- 18507963 TI - [Surgical treatment of well differentiated neuroendocrine tumours of the lung]. AB - The bronchial tree represents the most frequent site of origin of carcinoids (around 25% of the total). The spectrum of differentiation of lung neuroendocrine tumors ranges from low-malignancy (carcinoids) to highly aggressive forms (small cell lung carcinoma) Diagnostic and therapeutic strategies therefore vary greatly. In well differentiated tumors (carcinoids) signs and symptoms are related to the airways obstruction in central forms, while peripheral forms are mostly discovered accidentally if asymptomatic. Clinical or subclinical paraneoplastic syndromes are associated in a minority of cases. Diagnostic work up includes CT multislice, bronchial endoscopy and Octreoscan with chest Single Photon Emission Computed Tomography (SPECT). Further contribute may be added by the (68), Ga-DOTA-D-Phe(1)-Tyr(3)-ocreotide (DOTATOC) and 5-hydroxytryptophan (5 HTP) PET-CT, at present available only in a few centres, and by endobronchial ultrasound (EBUS), fluorescence bronchoscopy and virtual bronchoscopy. Surgery is the treatment of choice, while medical therapy is useful to treat the hypersecretion in paraneoplastic syndromes and to control tumor proliferation in metastatic or/and inoperable disease. PMID- 18507965 TI - The Greater Southern Area Health Service Tobacco Control Plan 2006-2009. AB - In response to the NSW Tobacco Action Plan 2005-2009, Greater Southern Area Health Service (GSAHS) has developed a local plan. This short report describes how activities promoted in the state plan were prioritised and six outcomes identified as the focus for the GSAHS Tobacco Control Plan 2006-2009. PMID- 18507966 TI - Provision of smoking care in NSW hospitals: opportunities for further enhancement. AB - The provision of smoking care, including the management of nicotine withdrawal and assistance with a quitting attempt, is identified as an important part of the overall care of hospitalised patients. Levels of smoking care delivery in hospitals have been less than optimal. Increasing this care across multiple facilities and units within NSW Health represents a significant challenge. This article examines levels of smoking care delivery in NSW hospitals, and research evidence and best practice recommendations to inform potential strategies to increase such care. It also reviews statewide initiatives implemented by NSW Health to enhance the delivery of smoking care and suggests further strategies that could facilitate this. PMID- 18507967 TI - Enabling the NSW health workforce to provide evidence-based smoking-cessation advice through competency-based training delivered via video conferencing. AB - Tobacco-related disease is estimated to cost the NSW health system more than $476 million in direct health care costs annually. Population-based smoking-cessation interventions, including brief intervention by health professionals, are effective and cost effective. As the prevalence of smoking in the general community declines, more highly dependent 'treatment-resistant' smokers may present a challenge to the health system. International guidelines recommend that health systems invest in training for health professionals in best practice smoking cessation. As part of the NSW Tobacco Action Plan 2005-2009, NSW Department of Health developed national competency standards in smoking cessation, designed learning and assessment materials and delivered training to more than 300 health professionals via video conference. Building the capacity of the NSW Health workforce to address smoking cessation as part of their routine practice is essential for addressing future challenges in tobacco control. PMID- 18507968 TI - Smoke free health care: an organisational change to increase effective intervention for tobacco. AB - In 1999, the NSW Health Smoke Free Workplace Policy directed that grounds of health sites would become smoke free, in addition to the existing policy requiring smoke-free buildings. This was one of the first attempts by any health service to exclude tobacco entirely from health sites. This task required the adoption of evidence-based management of tobacco dependence and changing the culture of smoking in the health service. There were many barriers to implementation. PMID- 18507969 TI - Tobacco and aboriginal people in NSW. AB - Tobacco use is a major cause of morbidity and mortality for Aboriginal people in NSW. Few interventions to reduce the harm resulting from tobacco use have been developed specifically for this population. However, brief interventions for smoking cessation, pharmacotherapies such as nicotine replacement therapy, bupropion and varenicline, quit groups and interventions aimed at reducing smoking by pregnant women and hospital inpatients are likely to be effective. Broader population interventions such as anti-tobacco advertising, price rises for tobacco products and prevention of sales to minors are also likely to be effective in reducing the harm resulting from tobacco use. PMID- 18507970 TI - Smokers respond to anti-tobacco mass media campaigns in NSW by calling the Quitline. AB - OBJECTIVE: One aim of the anti-tobacco mass media campaigns in NSW is to increase the numbers of smokers calling the Quitline to seek assistance to quit. METHODS: Investigation of the relationship between the weight of television advertising (Target Audience Rating Points: TARPs) and number of calls to the Quitline. RESULTS: There is a high correlation between weekly Quitline calls and TARPs. CONCLUSIONS: Anti-tobacco mass media campaign advertising that promotes the Quitline number encourages smokers to seek assistance to quit. PMID- 18507971 TI - The Mull Hypothesis: is cannabis use contributing to high tobacco use prevalence among young North Coast males? AB - Men aged 25-34 years, in North Coast NSW, have higher documented smoking rates than elsewhere in the state. The present paper explores potential causes of elevated smoking rates in this population and proposes that tobacco dependence resulting from 'mulling' (mixing) cannabis with tobacco may be contributing. PMID- 18507972 TI - Communicable Diseases Report, NSW, January and February 2008. AB - For updated information, including data and facts on specific diseases, visit (http://www.health.nsw.gov.au) and click on Infectious Diseases. PMID- 18507984 TI - Laudatio: professor Igor Buchwalow on the occasion of his 70th birthday (Gerhard Domagk Institute for Pathology, University of Munster, Germany). PMID- 18508008 TI - Retraction notice to "Simple, selective and sensitive spectrophotometric determination of fenitrothion using novel chromogenic reagent" [J. Hazard. Mater. 146 (1/2) (2007) 51-57]. PMID- 18508017 TI - Looking at health and teens. PMID- 18508021 TI - ODs find important role in care for patients with neurologic conditions. PMID- 18508022 TI - Highlights: 2007 American Optometric Association New Technology Survey. PMID- 18508023 TI - The 2007 Eyeglasses and Eye Care Services Index. PMID- 18508024 TI - Branding can help set a practice apart. PMID- 18508025 TI - The National Provider Identifier is here. PMID- 18508026 TI - Three steps to IACS-PC registration. PMID- 18508027 TI - Resistance to pyrethroid insecticides in Culex pipiens pipiens (Diptera: Culicidae) from Tunisia. AB - Resistance toward 2 pyrethroid insecticides (permethrin and deltamethrin), and an organochloric one (DDT) was analyzed in 18 samples of Culex pipiens pipiens common mosquitoes collected from different Tunisian areas between March 2002 and November 2005. Bioassays were performed over different larvae samples. The recorded mortalities, after 24h exposure to increased doses of insecticides, were compared to those obtained on a susceptible reference strain (S-LAB). All samples were resistant to permethrin. However, a large variation in the tolerance to this insecticide was observed between samples. Resistance ratio levels at LC50 (RR50) for the analyzed samples were ranged from 1.5 to 9092 folds. For deltamethrin, 14 among 17 studied samples were resistant. RR50, in the resistant samples, were ranged from 3 to 453 folds. Resistance to the two tested pyrethroids and DDT were correlated. The use of synergists showed that the cytochrome P450-dependent monooxygenases had a major contribution to the permethrin and deltamethrin resistance and that the esterases (and/or glutathione-S-transferases) had only a minor contribution. Results were discussed in relation to resistance mechanisms and mosquito's control. PMID- 18508028 TI - Static and dynamic postural loadings during computer work in females: Sitting on an office chair versus sitting on an exercise ball. AB - Seated computer work results in prolonged static loading, which has been associated with the development of musculoskeletal disorders. A popular alternative to sitting on an office chair while performing computer work is to sit on an exercise ball. Sitting on an exercise ball might affect static and dynamic aspects of working posture. We monitored posture, muscle activation and spinal shrinkage in 10 females performing a 1-h typing task, while sitting on an office chair with armrests and while sitting on an exercise ball. Sitting on an exercise ball resulted in 33% more trunk motion and in 66% more variation in lumbar EMG. Both of these findings can be considered to be an advantage for the exercise ball. However, the fifth percentile and average lumbar EMG were also higher when sitting on an exercise ball, with 38% and 78%, respectively. In addition, more spinal shrinkage occurred when sitting on an exercise ball than when sitting on an office chair. Arm flexion was reduced, but trapezius activation was unaffected when sitting on an exercise ball. It is concluded that the advantages with respect to physical loading of sitting on an exercise ball may not outweigh the disadvantages. PMID- 18508029 TI - Influence of arm and wrist support on forearm and back muscle activity in computer keyboard operation. AB - The effects of forearm and wrist supports on the upper extremity postures in computer keying tasks and associated EMG activity of arm and back muscles were examined (N = 8). Four positions were forearms unsupported (floating) and supported, wrists supported by bead packed (WR1) and gel-filled (WR2) wrist rest. The right and left elbow extensions were 65 degrees and 68 degrees , respectively, in unsupported forearms. Bilateral elbow extension increased with the forearm/wrist supports and mostly, the elbow was maintained at around 90 degrees or greater. The wrist extension decreased with forearm/wrist supports over the unsupported condition. The forearm support significantly reduced the activity of forearm extensor digitorum, i.e., right (F((1, 47)) = 12.19, p < 0.01) and left (F((1, 47)) = 5.38, p < 0.05) and upper trapezius muscles over the floating posture. Wrist rests, however, increased load on the upper trapezius; the activities of flexor digitorum superficialis and erector spinae were close to the resting EMG activity for both forearm and/or wrist support. The type of wrist rests was a concern and this study indicated that the gel filled wrist rest was advantageous in reducing the forearm muscle load, in comparison to the bead packed cushions. PMID- 18508030 TI - Expression of proinflammatory cytokines in osteoarthritis of the temporomandibular joint. AB - OBJECTIVE: This study reports the expression of proinflammatory cytokines in temporomandibular joint (TMJ) of patients affected with temporomandibular osteoarthritis (OA). DESIGN: In twelve OA of the TMJ (OA-TMJ) affected patients and in six healthy volunteer subjects studied as control, the expression of IL1beta (interleukin-1beta), IL2, IL4, IL5, IL6, IL10, IL12p35, IL12p40, IL17, IFNgamma (interferon-gamma), TNFalpha (tumor necrosis factor-alpha), and TNFbeta mRNAs was evaluated. Using quantitative real-time RT-PCR technique, the cytokine levels, reported as Ct (cycle threshold), DeltaCt (Ct cytokine-Ct 18S rRNA) and RQ (relative quantification), in patient and control groups were compared. RESULTS: Expression of IL1beta, IL2, IL12p35, IL12p40, IL17, TNFalpha, TNFbeta, and IFNgamma mRNAs was significantly higher in patients as compared with controls. In particular, IL12 was the predominant cytokine expressed in patients (IL12p35 RQ=30.2 and IL12p40 RQ=29.0). Conversely, IL10 mRNA levels were higher in controls (RQ=1.8). CONCLUSIONS: These data suggest that not only IL1beta, IFNgamma, and TNFalpha but also IL10, IL12, and IL17 are involved in the OA-TMJ pathogenesis. Furthermore, an inflammatory response characterised by the predominant expression of IL12 mRNA and down-regulated expression of IL10 mRNA is associated with the degenerative changes observed in OA-TMJ. PMID- 18508031 TI - Carnosic acid and carnosol potently inhibit human 5-lipoxygenase and suppress pro inflammatory responses of stimulated human polymorphonuclear leukocytes. AB - Carnosic acid (CA) and carnosol (CS) are phenolic diterpenes present in several labiate herbs like Rosmarinus officinalis (Rosemary) and Salvia officinalis (Sage). Extracts of these plants exhibit anti-inflammatory properties, but the underlying mechanisms are largely undefined. Recently, we found that CA and CS activate the peroxisome proliferator-activated receptor gamma, implying an anti inflammatory potential on the level of gene regulation. Here we address short term effects of CA and CS on typical functions of human polymorphonuclear leukocytes (PMNL). We found that (I), CA and CS inhibit the formation of pro inflammatory leukotrienes in intact PMNL (IC(50)=15-20 microM [CA] and 7 microM [CS], respectively) as well as purified recombinant 5-lipoxygenase (EC number 1.13.11.34, IC(50)=1 microM [CA] and 0.1 microM [CS], respectively), (II) both CA and CS potently antagonise intracellular Ca(2+) mobilisation induced by a chemotactic stimulus, and (III) CA and CS attenuate formation of reactive oxygen species and the secretion of human leukocyte elastase (EC number 3.4.21.37). Together, our findings provide a pharmacological basis for the anti-inflammatory properties reported for CS- and CA-containing extracts. PMID- 18508032 TI - Oxaliplatin activity in selected and unselected human ovarian and colorectal cancer cell lines. AB - Oxaliplatin is used for treatment of colon cancer in combination with 5 fluorouracil or irinotecan. Oxaliplatin has similar, but also different resistant mechanisms as cisplatin. We studied the activity of oxaliplatin in ovarian and colon cancer cells with different resistance patterns to cisplatin. The 40-fold cisplatin-resistant cell line ADDP was only 7.5-fold resistant to oxaliplatin. The gemcitabine-resistant AG6000 cell line, 9-fold resistant to cisplatin, was not cross-resistant. LoVo-175X2, with mutant p53 showed no resistance compared to the empty vector control. However, LoVo-Li, with inactive p53, was 3.6-fold resistant corresponding to decreased accumulation and Pt adducts. Accumulation and DNA adducts formation showed no significant correlation with oxaliplatin sensitivity. Cell cycle distribution after exposure to oxaliplatin showed arrest in G2/M (A2780) or in S-phase (LoVo-92) for wt-p53 cells. ADDP and LoVo-Li showed G1 arrest followed by S-phase arrest and no changes in distribution, respectively. The cell cycle related proteins Cyclins A and B1 and (p)CDC25C were marginally affected by oxaliplatin. Expression of hCTR1 was decreased in ADDP, LoVo-Li and AG6000, OCT1 decreased in ADDP and AG6000 and OCT3 in LoVo-175X2, compared to the parental cell lines. In ADDP and LoVo-175X2 ATP7A and B were decreased but were increased in AG6000. From this study it can be concluded that changes in cell cycle distribution were cell line dependent and not related to changes in expression of Cyclin A or B1. Oxaliplatin accumulation was related to hCTR1 and, at low concentration, ATP7A to DNA adducts formation while the retention was related to hCTR1, OCT2 and ATP7B. PMID- 18508033 TI - GSK3beta and endoplasmic reticulum stress mediate rotenone-induced death of SK-N MC neuroblastoma cells. AB - Rotenone, an environmental toxin that inhibits mitochondrial complex I, has been used to induce experimental Parkinsonism in animals and cell cultures. We investigated the mechanism underlying rotenone-induced death of SK-N-MC neuroblastoma cells. Rotenone-induced cell death preceded intracellular accumulation of reactive oxygen species, and antioxidants failed to protect cells, indicating that oxidative stress was minimally involved in rotenone induced death of SK-N-MC cells. Glycogen synthase kinase 3beta (GSK3beta), a multifunctional serine/threonine kinase, has been implicated in the pathogenesis of neurodegeneration. We showed that rotenone activated GSK3beta by enhancing its phosphorylation at tyrosine 216 while inhibiting phosphorylation at serine 9. Inhibitors of GSK3beta and dominant negative (kinase deficient) GSK3beta partially protected SK-N-MC cells against rotenone cytotoxicity. Rotenone also induced endoplasmic reticulum (ER) stress which was evident by an increase in phosphorylation of PERK, PKR, and eIF2alpha as well as the expression of GRP78. Rotenone had a modest effect on the expression of CHOP. An eIF2alpha siRNA significantly reduced rotenone cytotoxicity. ER stress was experimentally induced by tunicamycin and thapsigargin, but tunicamycin/thapsigargin did not activate GSK3beta in SK-N-MC cells. Down-regulation of eIF2alpha also offered partial protection against rotenone cytotoxicity. Combined treatment of GSK3beta inhibitors and eIF2alpha siRNA provided much greater protection than either treatment alone. Taken together, the results suggest that GSK3beta activation and ER stress contribute separately to rotenone cytotoxicity. PMID- 18508034 TI - Dimethylcelecoxib inhibits prostaglandin E2 production. AB - Dimethylcelecoxib (DMC), a derivative of celecoxib, has been developed to distinguish between the COX-dependent and COX-independent anti-carcinogenic effects of celecoxib. Although DMC has been shown to have no COX-inhibitory activity, it is important to ensure that DMC has no other influence on prostaglandin production. Interestingly, in this study we show that DMC inhibits PGE(2) production in vitro in the low micromolar range in different cancer cell lines. This effect can be at least partly explained by our findings that DMC inhibits microsomal prostaglandin E synthase-1 (mPGES-1) activity in a cell-free assay. Moreover, it prevents mPGES-1 up-regulation after stimulation of HeLa cells with IL-1beta and TNFalpha. Conversely, DMC has no effect on the expression levels of COX-1, COX-2, cytosolic PGES (cPGES) or mPGES-2 in these cells. However, in the cell-free assay DMC inhibits mPGES-1 to a maximum of 65% only and concentrations needed for inhibition of mPGES-1 activity are about 10-fold higher than needed for inhibition of PGE(2) production in cell culture. This suggests that DMC also has an impact on other proteins involved in PGE(2) production. In cell culture experiments the anti-proliferative effect of DMC, measured by the WST-1 assay, seems not to be dependent on PGE(2) inhibition, as DMC was equally effective in unstimulated HeLa cells as well as in stimulated HeLa cells, and the addition of external PGE(2) did not reverse the anti-proliferative effect of DMC in HCA-7 cells. We conclude that DMC is not a suitable non-prostaglandin inhibiting control substance for research purposes. PMID- 18508035 TI - Histone H2AX phosphorylation as a molecular pharmacological marker for DNA interstrand crosslink cancer chemotherapy. AB - The aims of this study were to investigate mechanisms of action involved in H2AX phosphorylation by DNA interstrand crosslinking (ICL) agents and determine whether gammaH2AX could be a suitable pharmacological marker for identifying potential ICL cellular chemosensitivity. In normal human fibroblasts, after treatment with nitrogen mustard (HN2) or cisplatin, the peak gammaH2AX response was detected 2-3 h after the peak of DNA ICLs measured using the comet assay, a validated method for detecting ICLs in vitro or in clinical samples. Detection of gammaH2AX foci by immunofluorescence microscopy could be routinely detected with 6-10 times lower concentrations of both drugs compared to detection of ICLs using the comet assay. A major pathway for repairing DNA ICLs is the initial unhooking of the ICL by the ERCC1-XPF endonuclease followed by homologous recombination. HN2 or cisplatin-induced gammaH2AX foci persisted significantly longer in both, ERCC1 or XRCC3 (homologous recombination) defective Chinese hamster cells that are highly sensitive to cell killing by ICL agents compared to wild type or ionising radiation sensitive XRCC5 cells. An advantage of using gammaH2AX immunofluorescence over the comet assay is that it appears to detect ICL chemosensitivity in both ERCC1 and HR defective cells. With HN2 and cisplatin, gammaH2AX foci also persisted in chemosensitive human ovarian cancer cells (A2780) compared to chemoresistant (A2780cisR) cells. These results show that gammaH2AX can act as a highly sensitive and general marker of DNA damage induced by HN2 or cisplatin and shows promise for predicting potential cellular chemosensitivity to ICL agents. PMID- 18508036 TI - Fate of rubrospinal neurons after unilateral section of the cervical spinal cord in adult macaque monkeys: effects of an antibody treatment neutralizing Nogo-A. AB - The present study describes in primates the effects of a spinal cord injury on the number and size of the neurons in the magnocellular part of the red nucleus (RNm), the origin of the rubrospinal tract, and evaluates whether a neutralization of Nogo-A reduces the lesioned-induced degenerative processes observed in RNm. Two groups of monkeys were subjected to unilateral section of the spinal cord affecting the rubrospinal tract; one group was subsequently treated with an antibody neutralizing Nogo-A; the second group received a control antibody. Intact animals were also included in the study. Counting neurons stained with a monoclonal antibody recognizing non-phosphorylated epitopes on neurofilaments (SMI-32) indicated that their number in the contralesional RNm was consistently inferior to that in the ipsilesional RNm, in a proportion amounting up to 35%. The lesion also induced shrinkage of the soma of the neurons detected in the contralesional RNm. Infusing an anti-Nogo-A antibody at the site of the lesion did not increase the proportion of SMI-32 positive rubrospinal neurons in the contralesional RNm nor prevent shrinkage. PMID- 18508037 TI - ATP-binding cassette transporter G4 is highly expressed in microglia in Alzheimer's brain. AB - Apolipoprotein E epsilon4 is an independent risk factor for Alzheimer's disease (AD) and is the main constituent of high-density lipoprotein (HDL) as a source of cholesterol in the brain. ATP-binding cassette transporter G4 (ABCG4) is one of the membrane cholesterol transporter which is implicated in HDL-mediated cholesterol efflux, but its precise localization and function in the brain has been unclear. In AD brain, ABCG4 protein was highly expressed in microglial cell that was closely located to senile plaques, whereas in non-neurological cases positive cells were not seen in cortical and nigral tissues. As well as the ABCG4 protein, ABCG4 mRNA signal was detected in microglial cell closely located to senile plaque of AD brain by in situ hybridization histochemistry. These results suggest that upregulated ABCG4 in microglia may accelerate the lipidation of apoE and HDL in the AD brain. This is the first report to show that ABCG4 is highly expressed in microglia on AD brain. PMID- 18508038 TI - Nuclear factor kappa-B mediates selective induction of neuronal nitric oxide synthase in astrocytes during low-level inflammatory stimulation with MPTP. AB - Recent advances in understanding the progression of Parkinson's disease (PD) implicate perturbations in astrocyte function and induction of constitutively expressed neuronal nitric oxide synthase (NOS1) in both human PD and in the MPTP model of the disease. Transcriptional regulation of NOS1 is complex but recent data suggest that nuclear factor kappa-B (NF-kappaB) is an important transcription factor involved in inducible expression of the gene. The data presented here demonstrate that mild activation of primary astrocytes with low or 'sub-optimal' concentrations of MPTP (1 microM) and the inflammatory cytokine tumor necrosis factor alpha (10 pg/ml) and interferon gamma (1 ng/ml) results in selective induction of Nos1 mRNA and protein, increased production of nitric oxide (NO), and a significant elevation in global protein nitration. This mild inflammatory stimulus also resulted in activation and recruitment of p65 to a putative NF-kappaB response element located in the Nos1 promoter region flanking exon 1. A role for NF-kappaB in MPTP-dependent induction of NOS1 was confirmed through overexpression of a mutant IkappaBalpha super repressor of NF-kappaB that prevented induction of NOS1. The data presented here thus demonstrate a role for NF-kappaB in selective induction of NOS1 during early inflammatory activation of astrocytes stimulated by low-dose MPTP and inflammatory cytokines. PMID- 18508039 TI - Chemo-enzymatic synthesis of C-9 acetylated sialosides. AB - A chemo-enzymatic synthesis of [(5-acetamido-9-O-acetyl-3,5-dideoxy-D-glycero alpha-D-galacto-2-nonulopyranosylonic acid)-(2-->3)-O-(beta-D-galactopyranosyl) (1-->3)-O-(2-acetamido-2-deoxy-alpha-D-galactopyranosyl)]-l-serine acetate (1) has been accomplished by a regioselective chemical acetylation of Neu5Ac (2) to give 9-O-acetylated sialic acid 3, which was enzymatically converted into CMP Neu5,9Ac(2) (4) employing a recombinant CMP-sialic acid synthetase from Neisseria meningitis [EC 2.7.7.43]. The resulting compound was then employed for the enzymatic glycosylation of the C-3' hydroxyl of chemically prepared glycosylated amino acid 10 using recombinant rat alpha-(2-->3)-O-sialyltransferase expressed in Spodooptera frugiperda [EC 2.4.99.4] to give, after deprotection of the N(alpha)-benzyloxycarbonyl (CBz)-protecting group of serine, target compound 1. The N(alpha)-CBz-protected intermediate 11 can be employed for the synthesis of glycolipopeptides for immunization purposes. PMID- 18508040 TI - A collaborative endeavor to design cholinesterase-based catalytic scavengers against toxic organophosphorus esters. AB - Wild-type human butyrylcholinesterase (BuChE) has proven to be an efficient bioscavenger for protection against nerve agent toxicity. Human acetylcholinesterase (AChE) has a similar potential. A limitation to their usefulness is that both cholinesterases (ChEs) react stoichiometrically with organophosphosphorus (OP) esters. Because OPs can be regarded as pseudo substrates for which the dephosphylation rate constant is almost zero, several strategies have been attempted to promote the dephosphylation reaction. Oxime mediated reactivation of phosphylated ChEs generates a turnover, but it is too slow to make pseudo-catalytic scavengers of pharmacological interest. Alternatively, it was hypothesized that ChEs could be converted into OP hydrolases by using rational site-directed mutagenesis based upon the crystal structure of ChEs. The idea was to introduce a nucleophile into the oxyanion hole, at an appropriate position to promote hydrolysis of the phospho-serine bond via a base catalysis mechanism. Such mutants, if they showed the desired catalytic and pharmacokinetic properties, could be used as catalytic scavengers. The first mutant of human BuChE that was capable of hydrolyzing OPs was G117H. It had a slow rate. Crystallographic study of the G117H mutant showed that hydrolysis likely occurs by activation of a water molecule rather than direct nucleophilic attack by H117. Numerous BuChE mutants were made later, but none of them was better than the G117H mutant at hydrolyzing OPs, with the exception of soman. Soman aged too rapidly to be hydrolyzed by G117H. Hydrolysis was however accomplished with the double mutant G117H/E197Q, which did not age after phosphonylation with soman. Multiple mutations in the active center of human and Bungarus AChE led to enzymes displaying low catalytic activity towards OPs and unwanted kinetic complexities. A new generation of human AChE mutants has been designed with the assistance of molecular modelling and computational methods. According to the putative water-activation mechanism of G117H BChE, a new histidine/aspartate dyad was introduced into the active center of human AChE at the optimum location for hydrolysis of the OP adduct. Additional mutations were made for optimizing activity of the new dyad. It is anticipated that these new mutants will have OP hydrolase activity. PMID- 18508041 TI - Coronary development is regulated by ATP-dependent SWI/SNF chromatin remodeling component BAF180. AB - Dissecting the molecular mechanisms that guide the proper development of epicardial cell lineages is critical for understanding the etiology of both congenital and adult forms of human cardiovascular disease. In this study, we describe the function of BAF180, a polybromo protein in ATP-dependent SWI/SNF chromatin remodeling complexes, in coronary development. Ablation of BAF180 leads to impaired epithelial-to-mesenchymal-transition (EMT) and arrested maturation of epicardium around E11.5. Three-dimensional collagen gel assays revealed that the BAF180 mutant epicardial cells indeed possess significantly compromised migrating and EMT potentials. Consequently, the mutant hearts form abnormal surface nodules and fail to develop the fine and continuous plexus of coronary vessels that cover the entire ventricle around E14. PECAM and *-SMA staining assays indicate that these nodules are defective structures resulting from the failure of endothelial and smooth muscle cells within them to form coronary vessels. PECAM staining also reveal that there are very few coronary vessels inside the myocardium of mutant hearts. Consistent with this, quantitative RT-PCR analysis indicate that the expression of genes involved in FGF, TGF, and VEGF pathways essential for coronary development are down-regulated in mutant hearts. Together, these data reveal for the first time that BAF180 is critical for coronary vessel formation. PMID- 18508042 TI - Identification of dkk4 as a target of Eda-A1/Edar pathway reveals an unexpected role of ectodysplasin as inhibitor of Wnt signalling in ectodermal placodes. AB - The development of epithelial appendages, including hairs, glands and teeth starts from ectodermal placodes, and is regulated by interplay of stimulatory and inhibitory signals. Ectodysplasin-A1 (Eda-A1) and Wnts are high in hierarchy of placode activators. To identify direct targets of ectodysplasin pathway, we performed microarray profiling of genes differentially regulated by short exposure to recombinant Eda-A1 in embryonic eda(-/-) skin explants. Surprisingly, there were only two genes with obvious involvement in Wnt pathway: dkk4 (most highly induced gene in the screen), and lrp4. Both genes colocalized with Eda-A1 receptor Edar in placodes of ectodermal organs. They were upregulated upon Edar activation while several other Wnt associated genes previously suggested as Edar targets were unaffected. However, low dkk4 and lrp4 expression was retained in the absence of NF-kappaB signalling in eda(-/-) hair placodes. We provide evidence that this expression was dependent on Wnt activity present prior to Eda A1/Edar signalling. Dkk4 was recently suggested as a key Wnt antagonist regulating lateral inhibition essential for correct patterning of hair follicles. Several pieces of evidence suggest Lrp4 as a Wnt inhibitor, as well. The finding that Eda-A1 induces placode inhibitors was unexpected, and underlines the importance of delicate fine-tuning of signalling during placode formation. PMID- 18508043 TI - Maternal Tgif is required for vascularization of the embryonic placenta. AB - The mammalian placenta is the site of exchange of nutrients and waste between mother and embryo. In humans, placental insufficiency can result in intrauterine growth retardation, perinatal death and spontaneous abortion. We show that in C57BL/6J mice a null mutation in the gene encoding the transcriptional corepressor, Tgif, causes placental defects. The major defects are decreased vascularization of the placenta, due to a decrease in the fetal blood vessels, and decreased expression of the gap junction protein Gjb2 (Cx26). These defects result in severe growth retardation in a proportion of Tgif null embryos in Tgif heterozygous mothers, and an overall growth delay in Tgif null animals. Placental defects are much more severe if the mother also completely lacks Tgif function, and placentas from heterozygous Tgif embryos are defective in a Tgif null mother. Embryo transfer experiments show that even the placenta from a wild type embryo is compromised in the absence of maternal Tgif. These results demonstrate that Tgif functions in the normal development of the placenta, and suggest a role for maternal factors in regulating the morphogenesis of embryonically-derived placental tissues. PMID- 18508044 TI - Donepezil attenuates excitotoxic damage induced by membrane depolarization of cortical neurons exposed to veratridine. AB - Long-lasting membrane depolarization in cerebral ischemia causes neurotoxicity via increases of intracellular sodium concentration ([Na+]i) and calcium concentration ([Ca2+]i). Donepezil has been shown to exert neuroprotective effects in an oxygen-glucose deprivation model. In the present study, we examined the effect of donepezil on depolarization-induced neuronal cell injury resulting from prolonged opening of Na+ channels with veratridine in rat primary-cultured cortical neurons. Veratridine (10 microM)-induced neuronal cell damage was completely prevented by 0.1 microM tetrodotoxin. Pretreatment with donepezil (0.1 10 microM) for 1 day significantly decreased cell death in a concentration dependent manner, and a potent NMDA receptor antagonist, dizocilpine (MK801), showed a neuroprotective effect at the concentration of 10 microM. The neuroprotective effect of donepezil was not affected by nicotinic or muscarinic acetylcholine receptor antagonists. We further characterized the neuroprotective properties of donepezil by measuring the effect on [Na+]i and [Ca2+]i in cells stimulated with veratridine. At 0.1-10 microM, donepezil significantly and concentration-dependently reduced the veratridine-induced increase of [Ca2+]i, whereas MK801 had no effect. At 10 microM, donepezil significantly decreased the veratridine-induced increase of [Na+]i. We also measured the effect on veratridine-induced release of the excitatory amino acids, glutamate and glycine. While donepezil decreased the release of glutamate and glycine, MK801 did not. In conclusion, our results indicate that donepezil has neuroprotective activity against depolarization-induced toxicity in rat cortical neurons via inhibition of the rapid influx of sodium and calcium ions, and via decrease of glutamate and glycine release, and also that this depolarization-induced toxicity is mediated by glutamate receptor activation. PMID- 18508045 TI - SA13353 (1-[2-(1-Adamantyl)ethyl]-1-pentyl-3-[3-(4-pyridyl)propyl]urea) inhibits TNF-alpha production through the activation of capsaicin-sensitive afferent neurons mediated via transient receptor potential vanilloid 1 in vivo. AB - Tumor necrosis factor-alpha (TNF-alpha) is known to play a crucial role in the pathogenesis of rheumatoid arthritis. In the present study, we demonstrate the effects of SA13353 (1-[2-(1-Adamantyl)ethyl]-1-pentyl-3-[3-(4 pyridyl)propyl]urea), a novel orally active inhibitor of TNF-alpha production, in animal models, and its mechanism of action on TNF-alpha production. SA13353 significantly inhibited lipopolysaccharide (LPS)-induced TNF-alpha production in a dose-dependent manner in rats. Moreover, SA13353 exhibited a binding affinity for the rat vanilloid receptor and increased neuropeptide release from the rat dorsal root ganglion neurons. However, its effects were blocked by pretreatment with the transient receptor potential vanilloid 1 (TRPV1) antagonist capsazepine. The ability of SA13353 and capsaicin to inhibit LPS-induced TNF-alpha production was eliminated by sensory denervation or capsazepine pretreatment in vivo. Although they inhibited LPS-induced TNF-alpha production in mice, these effects were not observed in TRPV1 knockout mice. SA13353 provoked the release of neuropeptides without nerve inactivation, even when chronically administered to rats. These results suggest that SA13353 inhibits TNF-alpha production through activation of capsaicin-sensitive afferent neurons mediated via TRPV1 in vivo. Post-onset treatment of SA13353 strongly reduced the hindpaw swelling and joint destruction associated with collagen-induced arthritis in rats. Thus, SA13353 is expected to be a novel anti-arthritic agent with a unique mechanism of action. PMID- 18508046 TI - Protective role of curcumin against nicotine-induced genotoxicity on rat liver under restricted dietary protein. AB - Nicotine, the well known addictive chemical of tobacco and active medication for several diseases, has proven to be a potential genotoxic compound. Although it is absorbed through lungs with smoking and mainly metabolized in liver, its effect on liver injuries is not clear. This study was designed to evaluate the genotoxicity of nicotine and corresponding the protective role of curcumin against nicotine on liver of female populations particularly who used tobacco but deprived of healthy diet. The effects were investigated by measurement of total DNA concentration of liver tissues and Comet assay of liver tissue DNA damage of female rats maintained under normal and restricted protein diets. Total DNA contents in the liver tissues were observed to decrease more significantly (P<0.001) by nicotine in both dietary conditions. Significant (P<0.01) increase of total DNA content in normal dietary condition and more significant (P<0.001) increase of total DNA content in protein restricted condition of the liver tissues were observed due to curcumin supplementations. Highly significant (P<0.001) DNA damages (37% in normal diet and 56% in protein restricted diet) of the liver tissues were observed due to nicotine treatment. Curcumin reduced the nicotine-induced DNA damage percentage of the liver tissues more significantly (P<0.001) in protein restricted condition. Curcumin proved its potential to function against genotoxic effect by reducing the DNA damage activity of nicotine and minimized the percentage of DNA damage (50-60%) in protein restricted dietary condition. The degree of nicotine-induced genotoxicity therefore can be effectively compensated by the protective effect of curcumin in protein stress condition. PMID- 18508047 TI - Both IKKalpha and IKKbeta are implicated in the arsenite-induced AP-1 transactivation correlating with cell apoptosis through NF-kappaB activity independent manner. AB - Arsenite has been well-proved to act as both an environmental carcinogen as well as a tumor therapeutic agent. AP-1 is one of the transcription factors that can be induced upon arsenite stimulation. However, the study on the mechanism and the function of the arsenite-induced AP-1 transactivation remains far complete. Here we demonstrated that high dose of arsenite induced apoptotic response in mouse fibroblasts correlating with AP-1 transactivation, which events were mediated by both IKKalpha and IKKbeta, two major protein kinases responsible for NF-kappaB activation. In addition, the regulatory effect of IKKs on the arsenite-induced AP 1 activation was delivered by sequential induction of GADD45alpha expression and the activation of MAPKK (MKK3/4/6) and MAPK (JNK and p38K)-dependent pathways. We further provided evidence that p50, but not p65 subunit of NF-kappaB, was involved in GADD45alpha induction and the subsequent MAPKK/MAPK/AP-1 activation under arsenite exposure, while functional NF-kappaB induced by arsenite stimulation consisted of p65 but not of p50 subunit. Therefore, we concluded that both IKKalpha and IKKbeta can mediate arsenite-induced AP-1 transactivation through NF-kappaB activity-independent manner. PMID- 18508048 TI - Expression and localisation of apical junctional complex proteins in lens epithelial cells. AB - The lens epithelium possesses an apical junctional complex (AJC) comprising adherens and tight junctions (AJs and TJs) and yet several key structural components and associated regulatory proteins have not been identified or localised in these cells. Here we determine the subcellular distribution of the archetypal TJ markers (ZO-1, claudin-1, and occludin) and TJ-associated cell polarity proteins (aPKC, Par3 and Par6beta) with AJ markers, E- and N-cadherin. As seen in other polarised epithelia, all these markers were located by confocal immunofluorescence microscopy to the apical ends of the lateral plasma membranes of bovine lens epithelial cells at sites of cell-cell interaction. Using immunoelectron microscopy, we show that ZO-1 concentrated at "kissing points" between neighbouring cells and these data, when taken in the context of our confocal immunofluorescence microscopy and blotting data, suggest the presence of TJs within the AJC. Likewise, immunogold labelling for E-cadherin identified AJs within these AJCs. We also report aPKC immunogold labelling localised to the AJC. These data show that the AJC of lens epithelial cells are a composite of TJs and AJs. PMID- 18508049 TI - Dietary restriction does not prevent nigrostriatal degeneration in the 6 hydroxydopamine model of Parkinson's disease. AB - Chronic reduction of the caloric intake is associated with extended lifespan, in rodents, and has been proposed to counteract neuronal loss in animal models of neurodegeneration. To test this hypothesis, we investigated the effect of dietary restriction (DR) in a rodent model of Parkinson's disease, based on the intrastriatal infusion of 6-hydroxydopamine. We could not confirm the neuroprotective effect of DR previously suggested: histological and behavioral measures indicated similar degrees of dopaminergic neuron loss in rats maintained on DR--for two or eight weeks prior to the lesion--or with free access to food. PMID- 18508050 TI - Echinococcus granulosus: cellular territories and morphological regions in mature protoscoleces. AB - Basic aspects of the generation, structure and function of Echinococcus granulosus protoscoleces are unknown. We review the work done on the structure and ultrastructure of the E. granulosus protoscolex and provide new data together with a comprehensive view of this form of the parasite. The surface, as observed by scanning electron microscopy, tightly correlates with five cellular territories characterized in the interior using light and transmission electron microscopy as well as a histochemical technique. Three of these territories are surrounded by a basal lamina that is also present in the internal side of the tegument, suggesting a complex internal organization. These cellular territories correlate with the expression of specific genes and the regionalization of DNA synthesis in protoscoleces. Additionally, a proposal to explain movements of the body of this form of the parasite in relation to the neck or to the germinal layer of the hydatid cyst is provided. PMID- 18508051 TI - The use of fluorescent fatty acid analogs as labels in trematode experimental infections. AB - We examined the utility of fluorescent fatty acid analog dyes for labeling larval trematodes to use in experimental infections. Our goals were to identify two dyes that label larval trematodes belonging to the species Maritrema novaezealandensis and Coitocaecum parvum, determine if the dyes influence survival and infectivity of larval trematodes and/or host mortality, and if larval trematodes labeled with alternative dyes could be distinguished post-infection. The two dyes tested, BODIPY FL C(12) and BODIPY 558/568 C(12), successfully labeled all treated larval trematodes, did not influence cercariae survival or infectivity, and did not influence host mortality in either host-parasite system. All larval parasites were fluorescent and distinguishable after 5 days in amphipod intermediate hosts. In addition, larval Acanthoparyphium sp. were strongly fluorescent with both dyes after 5 weeks within cockle hosts. This method should be extremely useful for experimental studies using trematode-host systems as models for addressing a range of ecological and evolutionary questions. PMID- 18508052 TI - Leishmania mexicana: participation of NF-kappaB in the differential production of IL-12 in dendritic cells and monocytes induced by lipophosphoglycan (LPG). AB - Dendritic cells (DC) and macrophages (Mphi) are well known as important effectors of the innate immune system and their ability to produce IL-12 indicates that they possess the potential of directing acquired immunity toward a Th1-biased response. Interestingly, the intracellular parasite Leishmania has been shown to selectively suppress Mphi IL-12 production and are DC the principal source of this cytokine. The molecular details of this phenomenon remain enigmatic. In the present study we examined the effect of Leishmania mexicana lipophosphoglycan (LPG) on the production of IL-12, TNF-alpha, and IL-10 and nuclear translocation of NF-kappaB. The results show that LPG induced more IL-12 in human DC than in monocytes. This difference was due in part to nuclear translocation of NF-kappaB, since LPG induced more translocation in DC than in monocytes. These results suggest that Leishmania LPG impairs nuclear translocation of NF-kappaB in monocytes with the subsequent decrease in IL-12 production. PMID- 18508053 TI - Intracellular signal transduction by the extracellular calcium-sensing receptor of Xenopus melanotrope cells. AB - The extracellular calcium-sensing receptor (CaR) is expressed in various types of endocrine pituitary cell, but the intracellular mechanism this G protein-coupled receptor uses in these cells is not known. In the present study we investigated possible intracellular signal transduction pathway(s) utilized by the CaR of the endocrine melanotrope cells in the intermediate pituitary lobe of the South African-clawed toad Xenopus laevis. For this purpose, the effects of various pharmacological agents on CaR-evoked secretion of radiolabeled secretory peptides from cultured melanotrope cells were assessed. CaR-evoked secretion, induced by the potent CaR agonist L-phenylalanine (L-Phe), could not be inhibited by cholera toxin, nor by NPC-15437 and PMA, indicating that neither G(s)/PKA nor G(q)/PKC pathways are involved. However, pertussis toxin (G(i/o) protein inhibitor), genistein (inhibitor of PTKs), wortmannin/LY-294002 (PI3-K inhibitor) and U-0126 (inhibitor of extracellular signal-regulated kinase, ERK) all substantially inhibited CaR-evoked secretion, indicating that the Xenopus melanotrope cell possesses a PI3-K/MAPK system that plays some role in CaR-signaling. Since no direct effect of L-Phe on ERK phosphorylation could be shown it is concluded that CaR must act primarily through another, still unknown, signaling pathway in Xenopus melanotropes. Our results indicate that the PI3-K/MAPK system has a facilitating effect on CaR-induced secretion, possibly by sensitizing the CaR. PMID- 18508054 TI - Industry-sponsored research. AB - Financial conflicts of interest are exceedingly common in biomedical research. Investigators with conflicts of interest are more likely to arrive at positive conclusions, perhaps as a result of biased study design, industry suppression of negative results, preferential funding by industry of projects that are likely to succeed, or biased interpretation of results on the part of investigators. Government and professional organisations have proposed guidelines for managing conflicts of interest, but in practice it is the policies of universities and medical journals that direct the actions of investigators. Academic researchers and the media have expressed concern about the influence of industry sponsorship on biomedical research, while industry is increasingly turning to private entities (such as contract research organisations) to conduct clinical trials. Research participants appear less concerned with conflicts of interest in biomedical research, perhaps due to a faith that such conflicts are being appropriately managed by institutions. After reviewing the literature, we provide recommendations for the ethical conduct of biomedical research in the presence of financial conflicts of interest. PMID- 18508055 TI - Translational research: from benchside to bedside. AB - Translation of the achievements of basic science into everyday clinical practice remains a major issue in contemporary medicine, and is addressed through a new discipline, translational research, which aims to bridge the gap between basic and clinical research. Translational research encompasses laboratory studies, clinical demands, public health and health management, policies and economics; it is crucial in the evolution of contemporary biomedical science; and its interventions follow the political-economic, ethical-social and educational scientific approaches. Translational research can progress through reorganisation of academic teams in a translational way. New academic posts translationally orientated are urgently needed, particularly in the field of trauma medicine, where lack of awareness of this new evolution is evident. PMID- 18508056 TI - Clinical research in trauma and orthopaedic surgery--call for action. PMID- 18508057 TI - Toxoplasma gondii infection in humans and animals in the United States. AB - This paper reviews clinical and asymptomatic Toxoplasma gondii infection in humans and other animals in the USA. Seroprevalence of T. gondii in humans and pigs is declining. Modes of transmission, epidemiology and environmental contamination with oocysts on land and sea are discussed. PMID- 18508059 TI - ApoE(-/-)/lysozyme M(EGFP/EGFP) mice as a versatile model to study monocyte and neutrophil trafficking in atherosclerosis. AB - OBJECTIVES: Intravital microscopy is a useful tool for studying leukocyte trafficking in atherosclerosis. However, distinction between various subclasses of leukocytes using this technology is lacking. Therefore, we generated ApoE(-/ )/Lysozyme M(EGFP/EGFP) mice and investigated whether targeted cell types could be visualized by in vivo microscopy and whether absence of lysozyme M will influence atherosclerosis. METHODS: We crossed male ApoE(-/-) mice with mice homozygous for a knock-in mutation of enhanced green fluorescent protein (EGFP) in the lysozyme M locus (Lys(EGFP/EGFP)) creating ApoE(-/-)/Lys(EGFP/EGFP) mice. Mice were sacrificed at the age of 26 weeks. Blood was collected for serum lipid analysis, differential white blood cell count and flow cytometry. Lesion area was determined on en face mounted aortas and sections from aortic roots were stained for immunohistochemistry. Atherosclerotic lesions were also studied by confocal- and intravital microscopy. RESULTS: Basic parameters, such as white blood cell count, cholesterol profile, lesion area and plaque composition was unaltered in ApoE(-/-)/Lys(EGFP/EGFP) mice compared to ApoE(-/-) mice. Fluorescent neutrophils and monocytes were clearly visualized by intravital fluorescence and confocal microscopy. Fluorescent cells were distributed primarily in the periphery of atherosclerotic lesions indicating a preference for recruitment in these areas. CONCLUSIONS: ApoE(-/-)/Lys(EGFP/EGFP) mice will serve as a useful model to study leukocyte trafficking in atherosclerosis and how different subsets of leukocytes influence atherogenesis. PMID- 18508060 TI - Increased expression of C-reactive protein and tissue factor in acute coronary syndrome lesions: Correlation with serum C-reactive protein, angioscopic findings, and modification by statins. AB - BACKGROUND: Serum C-reactive protein (CRP) is a strong risk predictor of cardiovascular events, and tissue factor (TF) plays a central role in thrombus formation of advanced atherosclerotic plaques. Aim of the present study was to quantify in situ CRP and TF in coronary atherectomy specimens associated with acute coronary syndromes (ACS) or stable angina (SA). In addition, the effect of statin treatment on both intimal determinants was analyzed. METHODS AND RESULTS: Serial sections from atherectomy probes retrieved from coronary primary target lesions of 42 ACS and 70 SA patients were examined for CRP and TF expression using immunostaining. CRP and TF intimal expression was consistently higher in ACS lesions and a positive correlation between both determinants was detected. In both subgroups intimal staining intensity of CRP but not TF was strongly associated with serum CRP levels. Using angioscopy, complex plaques revealed a higher intimal CRP and TF expression than white/yellow plaques. Both CRP and TF were consistently lower expressed in target lesions of patients with pre-existing statin treatment. CONCLUSIONS: CRP and TF expression is markedly increased in plaques derived from patients with ACS as compared to SA patients. Statin treatment appears to reduce vascular expression of CRP and TF. PMID- 18508061 TI - Re: Botox produces functional weakness in non-injected muscles adjacent to the target muscle. PMID- 18508062 TI - Functional analysis of Fontan energy dissipation. AB - We formalize the hydrodynamic energy dissipation in the total cavopulmonary connection (TCPC) using dimensional analysis and examine the effect of governing flow variables; namely, cardiac output, flow split, body surface area, Reynolds number, and certain geometric characteristics. A simplistic and clinically useful mathematical model of the dependence of energy dissipation on the governing variables is developed. In vitro energy loss data corresponding to six patients' anatomies validated the predicted dependency of each variable and was used to develop a predictive, semi-empirical energy dissipation model of the TCPC. It is shown that energy dissipation is a cubic function of pulmonary flow split in the physiological range. Furthermore, non-dimensional energy dissipation, which is a measure of resistance of the connection, is dependent on Reynolds number and geometrical factors alone. Non-dimensional energy dissipation decreases with Reynolds number as Re(-0.25) (R(2)>0.95). In addition, for high Reynolds numbers, within physiological exercise limits, dissipation strongly correlates to minimum PA area as a power law decay with an exponent of -5/4 (R(2)>0.88). This study presents a simple analytical form of energy dissipation rate in complex patient specific TCPCs that accurately captures the effect of cardiac output, flow split, body surface area, Reynolds number, and pulmonary artery size within physiological limits. Further studies with larger sample sizes are necessary for incorporating finer geometrical parameters such as vessel curvatures and offsets. PMID- 18508063 TI - Time-related PDL: viscoelastic response during initial orthodontic tooth movement of a tooth with functioning interproximal contact-a mathematical model. AB - Most anteroposterior orthodontic movements of posterior teeth have to overcome the "resistance" of adjacent teeth with functioning interproximal contacts. The aim of this study was to develop a mathematical model describing initial posterior tooth movement associated with functioning interproximal contacts in relation to the viscoelastic mechanical behavior of the human periodontal ligament (PDL). A linear viscoelastic 2D mathematical model was modified to depict tipping movement around the center of rotation (C(rot)) of a premolar where tipping is restrained by adjacent teeth. Equilibrium equations were applied taking into account the sagittal moment developed around the C(rot). The constants of the model were analyzed and applied to a numerical model that can simulate short-term tooth creep movement caused by a tipping force. Changes in force magnitude (0.5-3N) and crown length (6-10mm) were analyzed until no movement was observed (steady state). Premolar displacement in contact with adjacent teeth showed a non-linear progression over time with an initial sharp tipping movement followed by a transient period of 2.6-7.1min. As tipping force increased the transient period increased. A similar but smaller effect was observed with an increase in crown length. The premolar initial displacement within the arch (3.2-19.5microm) is about seven-fold smaller than retraction/protraction movement of an incisor. These suggest reduction in tooth displacement when functioning interproximal contact is present and clinically recommend establishing a space in the direction of tooth displacement before tooth movement. PMID- 18508064 TI - Chromatographic performance of macroporous cellulose-tungsten carbide composite beads as anion-exchanger for expanded bed adsorption at high fluid velocity. AB - Macroporous cellulose-tungsten carbide composite beads was designed and prepared as an anion-exchanger for expanded bed adsorption (EBA). The wet density of composite beads was adjusted at the range of 1.2-2.4 g/ml with the control of tungsten carbide addition, and optimized for EBA at high fluid velocity. The results indicated that the wet density of composite beads could increase linearly with the increase of tungsten carbide addition, meanwhile other physical properties, such as size, porosity, specific surface area, mean pore diameter, etc., were hardly or slightly influenced. The composite beads were coupled with diethylaminoethyl (DEAE) as an anion-exchanger for EBA. The expansion characteristics in expanded bed were investigated and sensitively changed as the wet density of composite beads, corresponding to tungsten carbide addition in the preparation. The relation among the operation fluid velocity, the ratio of tungsten carbide to cellulose viscose in the preparation and the expansion factor was found, which could be used to predict the operation velocity of composite beads with varying tungsten carbide addition. The liquid mixing in expanded bed was also tested and showed good bed stability for EBA processes. With the adsorption equilibrium experiments, the saturated adsorption capacity of bovine serum albumin could reach 68.7 mg/g adsorbents (equal to 97.1 mg/ml adsorbents). The ratio of Q(10%) (the dynamic adsorption at 10% breakthrough) in expanded bed to packed bed could reach more than 90% for the fluid velocity of 500 cm/h, even 77.1% for the fluid velocity as high as 900 cm/h. The chromatographic results demonstrated that the composite beads prepared are suitable for EBA applications at high fluid velocity. PMID- 18508065 TI - Determination of free medium-chain fatty acids in beer by stir bar sorptive extraction. AB - Free medium-chain fatty acids in beer originate from raw materials, mainly from the fermentation activity of yeasts, and can influence beer taste, vitality of yeasts and also the foam stability of beer. This study presents the development of the method for the determination of free fatty medium-chain acids including caproic acid, caprylic acid, capric acid and lauric acid in beer or wort using stir bar sorptive extraction (SBSE). The combination of this extraction technique with solvent back extraction of the extracted analytes and subsequent gas chromatographic analysis with flame ionization detection was used for the determination of these compounds. The influences of different solvent back solutions, sampling time, solvent back extraction times and different contents of ethanol were studied. The method had high repeatability (RSD <6.7%), good linearity (the correlation coefficients were higher than 0.9963 for quadratic curves over the concentration range 0.5-8.0mg/l) and recoveries 57-89%. PMID- 18508066 TI - Comparative metabolomic study of transgenic versus conventional soybean using capillary electrophoresis-time-of-flight mass spectrometry. AB - In this work, capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF MS) is proposed to identify and quantify the main metabolites found in transgenic soybean and its corresponding non-transgenic parental line both grown under identical conditions. The procedure includes optimization of metabolites extraction, separation by CE, on-line electrospray-TOF-MS analysis and data evaluation. A large number of extraction procedures and background electrolytes are tested in order to obtain a highly reproducible and sensitive analytical methodology. Using this approach, a large number of metabolites were tentatively identified based on the high mass accuracy provided by TOF-MS analyzer, together with the isotopic pattern and expected electrophoretic mobility of these compounds. In general, the same metabolites and in similar amounts were found in the conventional and transgenic variety. However, significant differences were also observed in some specific cases when the conventional variety was compared with its corresponding transgenic line. The selection of these metabolites as possible biomarkers of transgenic soybean is discussed, although a larger number of samples need to be analyzed in order to validate this point. It is concluded that metabolomic procedures based on CE-MS can open new perspectives in the study of transgenic foods in order to corroborate (or not) the equivalence with their conventional counterparts. PMID- 18508067 TI - Enantiomer identification in the flavour and fragrance fields by "interactive" combination of linear retention indices from enantioselective gas chromatography and mass spectrometry. AB - This study describes the development of a gas chromatography-mass spectrometry (GC-MS) library to identify optically active compounds in the flavour and fragrance field using enantioselective GC with cyclodextrin derivatives (CDs) as chiral selectors in combination with MS. The library operates on the "interactive" combination of linear retention indices (I(T) values) in parallel to MS spectra, so as to identify enantiomers reliably and to measure EE and/or ER unequivocally. Since MS is not a selective probe to discriminate optical isomers, mass spectra (or diagnostic ions in SIM mode) are used to locate the enantiomer(s) in the chromatogram, and I(T) values to identify it(them) safely and reliably in particular in complex mixtures. The library has been built up through the following steps: Some applications of the library are also reported. (a) Selection of CD derivatives able to cover a wide range of racemate separations. Four cyclodextrin derivatives were used: 2,6-di-O-methyl-3-O-pentyl beta-CD, 2,3-di-O-methyl-6-O-tert-butyldimethylsilyl-beta-CD, 2,3-di-O-ethyl-6-AO tert-butyldimethylsilyl-beta-CD, and 2,3-di-O-acetyl-6-O-tert-butyldimethylsilyl beta-CD. (b) Determination of the analytes' I(T) values and evaluation of their stability and reliability at both intra- and inter-laboratory level. (c) Determination of the range within which the I(T) of an enantiomer has to be correctly identified, i.e. determination of a common retention index allowance (RIA). (d) Construction of the library, at the moment comprising the enantiomers of 134 racemates. A record has been attributed to each enantiomer including I(T) values determined on the four CD coated columns, mass spectrum, IUPAC chemical names, CAS numbers, molecular weight, and, when separated, racemate enantiomer resolution on the CD investigated. Some applications of the library are also reported. PMID- 18508068 TI - Ion-trap versus time-of-flight mass spectrometry coupled to capillary electrophoresis to analyze biogenic amines in wine. AB - In this work, two different capillary electrophoresis-mass spectrometry (CE-MS) methods, namely, capillary electrophoresis-ion-trap mass spectrometry (CE-IT-MS) and capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF-MS), applied to analyze biogenic amines in wine samples are investigated. A group of five amines was selected as case study (namely, putrescine, cadaverine, histamine, phenylethylamine and tyramine) since they are the most frequently biogenic amines found in wines. The possibilities of both instruments in terms of sensitivity, selectivity and quantitation during the determination of the mentioned biogenic amines in wine samples was studied and their results corroborated by HPLC. After optimization of the analytical conditions, CE-IT-MS and CE-TOF-MS allowed the identification of biogenic amines in wines without any previous treatment except diluting 1:1 with water and filtering. Biogenic amines were determined in three red wines and one white wine showing, as expected, a higher concentration in red wines. Moreover, CE-IT-MS and CE-TOF-MS were compared regarding their capacity to detect other biogenic amines different to the selected ones in wine samples, showing CE-TOF-MS a much better capability (i.e., putrescine, cadaverine, histamine, phenylethylamine, tyramine, triptamine, spermidine, spermine, ethanolamine and isoamylamine were identified by CE-TOF-MS in a single analysis). Moreover, CE-TOF-MS allowed the quantitation of biogenic amines with limits of detection as low as 10 ng/mL, comparable to those obtained using HPLC with fluorescence detection, without any previous derivatization step and with analysis times fivefold faster (40 min by HPLC and 8 min by CE-TOF-MS). PMID- 18508069 TI - Interaction of supercritical CO2 with alkyl-ammonium organoclays: changes in morphology. AB - This paper investigates the influence of supercritical carbon dioxide on the morphology and surface chemistry of three organic modified montmorillonite species. Alkyl based quaternary ammonium surfactants with differing numbers of chains attached, were chosen to vary the degree of CO(2)-philicity exhibited by the organoclay. In a high pressure batch vessel, the different organoclays were suspended in the supercritical solvent at temperatures of 50 and 200 degrees C and pressures of 7.6 and 9.6 MPa and then removed after de-pressurization at 0.2 or 4.8 MPa/s. The structures of these treated clays were characterized by X-ray diffraction (XRD), differential scanning microscopy (DSC), and thermogravimetric analysis (TGA), and their chemical properties were analyzed by various methods including atomic absorption spectroscopy, and water uptake measurement. Solute solvent interactions plasticized the organic medium while suspended in the supercritical fluid, which resulted in greater chain mobility and further cation exchange. The results indicate that intercalated surfactants exhibiting a paraffin complex arrangement were most likely to experience significant basal expansion, provided the tilt angle was not already close to being perpendicular to the silicate surface. At the lower processing temperature condition, the chemistry of the clay surface was notably altered by the CO(2) associations with the Lewis acid/base sites, which significantly reduced the moisture adsorption capacity of the material. For those organoclays demonstrating basal expansion, it was noted that the resulting particle size was increased due to enhanced porosity. PMID- 18508070 TI - Viscoelastic properties of adsorbed and cross-linked polypeptide and protein layers at a solid-liquid interface. AB - The real-time changes in viscoelasticity of adsorbed poly(L-lysine) (PLL) and adsorbed histone (lysine rich fraction) due to cross-linking by glutaraldehyde and corresponding release of associated water were investigated using a quartz crystal microbalance with dissipation monitoring (QCM-D) and attenuated total reflection Fourier transform infrared spectroscopy (ATR/FTIR). The kinetics of PLL and histone adsorption were measured through changes in mass adsorbed onto a gold-coated quartz surface from changes in frequency and dissipation and using the Voigt viscoelastic model. Prior to cross-linking, the shear viscosity and shear modulus of the adsorbed PLL layer were approximately 3.0 x 10(-3) Pas and approximately 2.5 x 10(5) Pa, respectively, while after cross-linking, they increased to approximately 17.5 x 10(-3) Pas and approximately 2.5 x 10(6) Pa, respectively. For the adsorbed histone layer, shear viscosity and shear modulus increased modestly from approximately 1.3 x 10(-3) to approximately 2.0 x 10(-3) Pas and from approximately 1.2 x 10(4) to approximately 1.6 x 10(4) Pa, respectively. The adsorbed mass estimated from the Sauerbrey equation (perfectly elastic) and the Voigt viscoelastic model differ appreciably prior to cross linking whereas after cross-linking they converged. This is because trapped water molecules were released during cross-linking. This was confirmed experimentally via ATR/FTIR measurements. The variation in viscoelastic properties increased substantially after cross-linking presumably due to fluctuation of the randomly cross-linked network structure. An increase in fluctuation of the viscoelastic properties and the loss of imbibed water could be used as a signature of the formation of a cross-linked network and the amount of cross-linking, respectively. PMID- 18508071 TI - Easy method for preparing nanocrystalline CdS hollow spheres using miniemulsion droplets as templates. AB - CdS hollow spheres with well-controlled morphology and uniform size were successfully prepared using a miniemulsion technique, in which miniemulsion droplets of isooctane prepared with dodecylmercaptane as a co-stabilizer were employed as templates. The SH groups of dodecylmercaptane generated S(-) ions under alkaline conditions, and further reacted with Cd(2+) ions to form cadmium thiolate around the droplets. The cadmium thiolate then reacted with S(2-) ions, stemming from Na(2)S9H(2)O, to directly form CdS hollow spheres owing to the evaporation of isooctane during the reaction and/or the subsequent drying process. No additional dissolution, calcination or additional surface modification of the templates was needed. The size of the hollow spheres could be tuned by altering the molar ratio of the anionic and non-ionic surfactants, while their shell thickness could be adjusted by changing the amount of co-stabilizer. PMID- 18508072 TI - Superhydrophobic cellulose nanocomposites. AB - Superhydrophobic cellulose nanocomposites were prepared using a multi-step nanoengineering process. The combination of different techniques made it possible to construct novel features at the ensuing surface, characterized by both an increase in its roughness induced by amorphous silica particles and a reduction in its energy insured by perfluoro moieties, giving rise to water contact angles approaching 150 degrees . The modification calls upon an aqueous LbL system followed by siloxane hydrolysis, both conducted at room temperature in air. Each modification was followed by scanning electron microscopy (SEM) and atomic force microscope (AFM). These original cellulose-silica-silane composite materials open the way to further valorisations of a ubiquitous renewable resource in applications such as water repellence and self-cleaning. PMID- 18508073 TI - Reduced aggregation and sedimentation of zero-valent iron nanoparticles in the presence of guar gum. AB - Injection of nanoscale zero-valent iron (NZVI) is potentially a promising technology for remediation of contaminated groundwaters. However, the efficiency of this process is significantly hindered by the rapid aggregation of the iron nanoparticles. The aim of this study was to enhance the colloidal stability of the nanoparticles through the addition of the "green" polymer guar gum. We evaluated the properties of guar gum and its influence on the surface properties, particle size, aggregation, and sedimentation of iron nanoparticles. Commercial iron nanoparticles were dispersed in guar gum solutions, and their aggregation and sedimentation behaviors were compared to those of bare iron nanoparticles and commercial nanoparticles modified with a biodegradable polymer (polyaspartate). High performance size exclusion chromatography, charge titration, and viscosity assessment showed that guar gum is a high molecular weight polymer which is nearly neutrally charged, rendering it suitable for steric stabilization of the iron nanoparticles. Electrophoretic mobility measurements demonstrated the ability of guar gum to adsorb on the nanoparticles, forming a slightly negatively charged layer. Dynamic light scattering experiments were conducted to estimate the particle size of the different nanoparticle suspensions and to determine the aggregation behavior at different ionic strengths. Guar gum effectively reduced the hydrodynamic radius of the bare nanoparticles from 500 nm to less than 200 nm and prevented aggregation of the nanoparticles even at very high salt concentrations (0.5 M NaCl and 3 mM CaCl(2)). Sedimentation profiles of the different nanoparticle suspensions confirmed the improved stability of the iron nanoparticles in the presence of guar gum. The results strongly suggest that guar gum can be used to effectively deliver stabilized zero-valent iron nanoparticles for remediation of contaminated groundwater aquifers. PMID- 18508074 TI - The ELISA Standard Save: calculation of sample concentrations in assays with a failed standard curve. AB - The enzyme-linked immunosorbent assay (ELISA) is often used to measure protein levels in plasma and other solutions. In order for the assay to be quantitative, a standard curve must be prepared for each assay. Technical blunders in preparing the standard curve can render otherwise representative sample values useless. In order to recover these data, a protocol has been developed whereby a new standard curve is generated using the DeltaOD values of the samples. This new standard is applied to the original plate in order to determine sample concentrations. To validate this method, sample concentrations obtained using an acceptable standard curve were plotted against those obtained using the new standard curve. Linear regression analysis showed a 1:1 correlation between concentrations, with r(2) values ranging from 0.98-0.99 and slopes ranging from 0.97-1.10. In this manner, data for these samples are preserved, saving the investigator the time and money involved in repeating experiments. PMID- 18508075 TI - Autoinhibition of human dicer by its internal helicase domain. AB - Dicer, a member of the ribonuclease III family of enzymes, processes double stranded RNA substrates into approximately 21- to 27-nt products that trigger sequence-directed gene silencing by RNA interference. Although the mechanism of RNA recognition and length-specific cleavage by Dicer has been established, the way in which dicing activity is regulated is unclear. Here, we show that the N terminal domain of human Dicer, which is homologous to DExD/H-box helicases, substantially attenuates the rate of substrate cleavage. Deletion or mutation of this domain activates human Dicer in both single- and multiple-turnover assays. The catalytic efficiency (k(cat)/K(m)) of the deletion construct is increased by 65-fold over that exhibited by the intact enzyme. Kinetic analysis shows that this activation is almost entirely due to an enhancement in k(cat). Modest stimulation of catalysis by the full-length Dicer enzyme was observed in the presence of the TAR-RNA binding protein, which physically interacts with the DExD/H-box domain. These results suggest that the DExD/H-box domain likely disrupts the functionality of the Dicer active site until a structural rearrangement occurs, perhaps upon assembly with its molecular partners. PMID- 18508076 TI - Kinetic partitioning between alternative protein-protein interactions controls a transcriptional switch. AB - Proteins can perform completely distinct functions in response to the particular partners that they bind to. Consequently, determination of the mechanism of functional regulation in such systems requires elucidation of the mechanism switching between binding partners. The central protein of the Escherichia coli biotin regulatory system, BirA, switches between its function as a metabolic enzyme or a transcriptional repressor in response to binding either the biotin carboxyl carrier protein subunit of acetyl-CoA carboxylase or a second BirA monomer. These two protein-protein interactions are structurally mutually exclusive. The results of earlier studies suggest that the system is regulated by kinetic partitioning between the two protein-protein interactions. In this work, sedimentation velocity was employed to monitor the partitioning directly. The results indicate similar equilibrium parameters governing formation of the two protein-protein interactions. Kinetic analysis of the sedimentation velocity data indicated that holoBirA dimerization is governed by very slow forward and reverse rate constants. The slow kinetics of holoBirA dimerization combined with fluctuations in the intracellular apoBCCP pool are critical determinants in partitioning BirA between its distinct biological functions. PMID- 18508077 TI - Recruitment of CREB1 and histone deacetylase 2 (HDAC2) to the mouse Ltbp-1 promoter regulates its constitutive expression in a dioxin receptor-dependent manner. AB - Latent TGFbeta-binding protein 1 (LTBP-1) is a key regulator of TGFbeta targeting and activation in the extracellular matrix. LTBP-1 is recognized as a major docking molecule to localize, and possibly to activate, TGFbeta in the extracellular matrix. Despite this relevant function, the molecular mechanisms regulating Ltbp-1 transcription remain largely unknown. Previous results from our laboratory revealed that mouse embryonic fibroblasts (MEF) lacking dioxin receptor (AhR) had increased Ltbp-1 mRNA expression and elevated TGFbeta activity, suggesting that AhR repressed Ltbp-1 transcription. Here, we have cloned the mouse Ltbp-1 gene promoter and analysed its mechanism of transcriptional repression by AhR. Reporter gene assays, AhR over-expression and site-directed mutagenesis showed that basal Ltbp-1 transcription is AhR dependent. Chromatin immunoprecipitation (ChIP) and RNA interference (RNAi) revealed that AhR regulates Ltbp-1 transcription by a mechanism involving recruitment of co-activators such as CREB1 and co-repressors such as HDAC2 to the Ltbp-1 promoter. In AhR-expressing (AhR+/+) MEF cells, the recruitment of HDAC1, 2 and 4 correlated with decreased K8H4 acetylation and impaired binding of pCREB(Ser133) to the Ltbp-1 promoter, likely maintaining a constitutive repressed state. AhR-/- MEF cells had the opposite pattern of HDACs and pCREB1(Ser133) binding to Ltbp-1 promoter, and therefore, over-expressed Ltbp-1 mRNA. In agreement, siRNA for HDAC2 increased Ltbp-1 expression and K8H4 acetylation in AhR+/+ but not in AhR-/- MEF cells. We suggest that HDAC2 binding keeps Ltbp-1 promoter repressed in AhR+/+ MEF cells, whereas in AhR-null MEF cells the absence of HDAC2 and the binding of pCREB(Ser133) allow Ltbp-1 transcription. Thus, epigenetics can contribute to constitutive Ltbp-1 repression by a mechanism requiring AhR activity. PMID- 18508079 TI - Immunogenic display of diverse peptides on virus-like particles of RNA phage MS2. AB - The high level of immunogenicity of peptides displayed in dense repetitive arrays on virus-like particles makes recombinant VLPs promising vaccine carriers. Here, we describe a platform for vaccine development based on the VLPs of RNA bacteriophage MS2. It serves for the engineered display of specific peptide sequences, but will also allow the construction of random peptide libraries from which specific binding activities can be recovered by affinity selection. Peptides representing the V3 loop of HIV gp120 and the ECL2 loop of the HIV coreceptor, CCR5, were inserted into a surface loop of MS2 coat protein. Both insertions disrupted coat VLP assembly, apparently by interfering with protein folding, but these defects were suppressed efficiently by genetically fusing coat protein's two identical polypeptides into a single-chain dimer. The resulting VLPs displayed the V3 and ECL2 peptides on their surfaces where they showed the potent immunogenicity that is the hallmark of VLP-displayed antigens. Experiments with random-sequence peptide libraries show the single-chain dimer to be highly tolerant of six, eight and ten amino acid insertions. MS2 VLPs support the display of a wide diversity of peptides in a highly immunogenic format, and they encapsidate the mRNAs that direct their synthesis, thus establishing the genotype/phenotype linkage necessary for recovery of affinity-selected sequences. The single-chain MS2 VLP therefore unites in a single structural platform the selective power of phage display with the high immunogenicity of VLPs. PMID- 18508078 TI - Ribonuclease A homologues of the zebrafish: polymorphism, crystal structures of two representatives and their evolutionary implications. AB - The widespread and functionally varied members of the ribonuclease A (RNase A) superfamily provide an excellent opportunity to study evolutionary forces at work on a conserved protein scaffold. Representatives from the zebrafish are of particular interest as the evolutionary distance from non-ichthyic homologues is large. We conducted an exhaustive survey of available zebrafish DNA sequences and found significant polymorphism among its four known homologues. In an extension of previous nomenclature, the variants have been named RNases ZF-1a-c,-2a-d,-3a-e and-4. We present the first X-ray crystal structures of zebrafish ribonucleases, RNases ZF-1a and-3e at 1.35-and 1.85 A resolution, respectively. Structure-based clustering with ten other ribonuclease structures indicates greatest similarity to mammalian angiogenins and amphibian ribonucleases, and supports the view that all present-day ribonucleases evolved from a progenitor with three disulphide bonds. In their details, the two structures are intriguing melting-pots of features present in ribonucleases from other vertebrate classes. Whereas in RNase ZF-1a the active site is obstructed by the C-terminal segment (as observed in angiogenin), in RNase ZF-3e the same region is open (as observed in more catalytically efficient homologues). The progenitor of present-day ribonucleases is more likely to have had an obstructive C terminus, and the relatively high similarity (late divergence) of RNases ZF-1 and-3 infers that the active site unblocking event has happened independently in different vertebrate lineages. PMID- 18508080 TI - New features of the ribosome and ribosomal inhibitors: non-enzymatic recycling, misreading and back-translocation. AB - We describe the optimization of a poly(Phe) synthesis system, the conditions of which have been applied for efficient translation of heteropolymeric mRNAs. Here we identify two parameters that are essential to obtain translation at efficiency and accuracy levels equivalent to those in vivo, viz., the fine-tuning of the energy-rich components with an acetyl-phosphate substrate for energy regeneration, as well as the ionic conditions. Applying this system revealed a number of new features: (i) 70S ribosomes are able to recycle within 300 s in a non-enzymatic fashion in the absence of tmRNA. This observation might explain the fact that a knockout of the tmRNA gene ssrA is not lethal for Escherichia coli cells in contrast to other bacterial strains, such as Bacillus subtilis. (ii) The high efficiency of the system was exploited to analyze the misincorporation of various amino acids (resolution limit=1:15,000). No misreading was observed at the middle codon position and only marginal effects were observed at the first one (even when misreading was artificially stimulated 20- to 30-fold), yielding an improved definition of the near-cognate and non-cognate aminoacyl-tRNAs. (iii) Aminoglycosides increase Phe and Lys incorporation about 2-fold in the presence of poly(U) or poly(UUC) and poly(A), respectively, and induce a back translocation (except hygromycin B) exclusively in the absence of EF-G*GTP, as do the non-related drugs viomycin and edeine. PMID- 18508081 TI - Cross-linking of phospholipid membranes is a conserved property of calcium sensitive synaptotagmins. AB - Synaptotagmins are vesicular proteins implicated in many membrane trafficking events. They are highly conserved in evolution and the mammalian family contains 16 isoforms. We now show that the tandem C2 domains of several calcium-sensitive synaptotagmin isoforms tested, including Drosophila synaptotagmin, rapidly cross link phospholipid membranes. In contrast to the tandem structure, individual C2 domains failed to trigger membrane cross-linking in several novel assays. Large scale liposomal aggregation driven by tandem C2 domains in response to calcium was confirmed by the following techniques: turbidity assay, dynamic light scattering and both confocal and negative stain electron microscopy. Firm cross linking of membranes was evident from laser trap experiments. High-resolution cryo-electron microscopy revealed that membrane cross-linking by tandem C2 domains results in a constant distance of approximately 9 nm between the apposed membranes. Our findings show the conserved nature of this important property of synaptotagmin, demonstrate the significance of the tandem C2 domain structure and provide a plausible explanation for the accelerating effect of synaptotagmins on membrane fusion. PMID- 18508082 TI - A betacellulin mutant promotes differentiation of pancreatic acinar AR42J cells into insulin-producing cells with low affinity of binding to ErbB1. AB - Betacellulin (BTC) is one of the members of the epidermal growth factor (EGF) ligand family of ErbB receptor tyrosine kinases. It is a differentiation factor as well as a potent mitogen. BTC promotes the differentiation of pancreatic acinar-derived AR42J cells into insulin-producing cells. It independently and preferentially binds to two type I tyrosine kinase receptors, the EGF receptor (ErbB1) and ErbB4. However, the physiochemical characteristics of BTC that are responsible for its preferential binding to these two receptors have not been fully defined. In this study, to investigate the essential amino acid residues of BTC for binding to the two receptors, we introduced point mutations into the EGF domain of BTC employing error-prone PCR. The receptor binding abilities of 190 mutants expressed in Escherichia coli were assessed by enzyme immunoassay. Replacement of the glutamic acid residue at position 88 with a lysine residue in BTC was found to produce a significant loss of affinity for binding to ErbB1, while the affinity of binding to ErbB4 was unchanged. In addition, the mutant of BTC-E/88/K showed less growth-promoting activity on BALB/c 3T3 cells compared with that of the wild-type BTC protein. Interestingly, the BTC mutant protein promoted differentiation of pancreatic acinar AR42J cells at a high frequency into insulin-producing cells compared with AR42J cells that were treated with wild-type BTC protein. These results indicate the possibility of designing BTC mutants, which have an activity of inducing differentiation only, without facilitating growth promotion. PMID- 18508083 TI - Structural basis of the honey bee PBP pheromone and pH-induced conformational change. AB - The behavior of insects and their perception of their surroundings are driven, in a large part, by odorants and pheromones. This is especially true for social insects, such as the honey bee, where the queen controls the development and the caste status of the other individuals. Pheromone perception is a complex phenomenon relying on a cascade of recognition events, initiated in antennae by pheromone recognition by a pheromone-binding protein and finishing with signal transduction at the axon membrane level. With to the objective of deciphering this initial step, we have determined the structures of the bee antennal pheromone-binding protein (ASP1) in the apo form and in complex with the main component of the queen mandibular pheromonal mixture, 9-keto-2(E)-decenoic acid (9-ODA) and with nonpheromonal components. In the apo protein, the C terminus obstructs the binding site. In contrast, ASP1 complexes have different open conformations, depending on the ligand shape, leading to different volumes of the binding cavity. The binding site integrity depends on the C terminus (111-119) conformation, which involves the interplay of two factors; i.e. the presence of a ligand and a low pH. Ligand binding to ASP1 is favored by low pH, opposite to what is observed with other pheromone-binding proteins, such as those of Bombyx mori and Anopheles gambiae. PMID- 18508084 TI - The CRF-like peptide urocortin produces a long-lasting recovery in rats made hemiparkinsonian by 6-hydroxydopamine or lipopolysaccharide. AB - We have recently observed that the corticotropin releasing factor related peptide urocortin (UCN) reverses key features of nigrostriatal neurodegeneration following intracerebral injection of either 6-hydroxydopamine (6-OHDA) or lipopolysaccharide (LPS). To determine the potential therapeutic utility of UCN here we have studied whether these effects are sustained for several weeks following peptide injection. In addition we have studied whether UCN still shows efficacy in rats with more pronounced nigrostriatal lesions. Rats were lesioned using 6-OHDA or LPS and injected with UCN either 7 or 14 days later. At different time points animals were tested for rotational behaviour (apomorphine, 0.5 mg/kg) and subsequently implanted with bilateral dialysis probes into the striata. The following day rats were dialysed to estimate extracellular striatal dopamine (DA) and then sacrificed for estimation of striatal tissue DA and subsequent immunohistochemistry of TH(+) cells in the substantia nigra (SN). Toxin treated rats given UCN 7 days later showed clear evidence of reduced nigrostriatal damage both 28 and 84 days following UCN compared with saline injection. In rats given UCN 14 days after toxin injection, by which time deficits were maximal, a restoration of nigrostriatal damage was observed. This suggests that UCN is able to elicit a sustained restoration of functional nigrostriatal integrity and has the ability to produce a recovery in severely lesioned rats. These findings suggest that stimulation of CRF (probably CRF(1)) receptors could have therapeutic utility in PD. PMID- 18508085 TI - Sleep disorders in Chinese patients with Parkinson's disease: validation study of a Chinese version of Parkinson's disease sleep scale. AB - To evaluate the Chinese version of the Parkinson's disease sleep scale (PDSS) as an instrument for measuring sleep disorders in Chinese patients with Parkinson's disease (PD). The objective of the present study was to carry out a metric analysis of a Chinese version of PDSS using a cross-sectional study of 126 patients with PD who participated in the study. Usual measures for PD patients including the Pittsburgh sleep quality index (PSQI), the Epworth sleepiness scale (ESS), the Geriatric Depression Scale (GDS), and the Hamilton Anxiety Scale (HAMA) were applied by neurologists. The intra-class correlation coefficient was 0.880, and test-retest reliability for total PDSS score was 0.914. The Mean total PDSS score was 118.38+/-26.07. There was a significant correlation between the PDSS and PSQI, between the PDSS and ESS, between the PDSS and GDS, between the PDSS and HAMA, between the PDSS and the disease durations, and between the PDSS and the LDE, respectively. The Chinese version of PDSS met some basic standards required for sleep disorders measures. It could lead to better understanding the sleep disorders of PD of China in future studies. PMID- 18508086 TI - Protective effect of apigenin on ovariectomy-induced bone loss in rats. AB - Recent studies have shown that apigenin not only inhibits bone resorption by osteoclasts but also induces osteoclast apoptosis. However, the influence of apigenin on osteoporosis in animals is relatively unknown. The purpose of this study was to examine the bone-protective effects of apigenin in estrogen deficient ovariectomized rats. Three-month-old female Sprague-Dawley rats were either sham-operated or ovariectomized and fed AIN-93G diet for 7 weeks to induce bone loss. To confirm bone loss, we used a newly developed non-invasive technique involving zoom-in micro-computed tomography. Apigenin was administered at a dose of 10 mg/kg three times a week for 15 weeks. Our results indicate that apigenin not only increased the mineral content and density of the trabecular bone at the neck of the left femur, but also decreased body weight and dietary consumption. Moreover, our biochemical results indicate that apigenin has a positive effect on bone turnover. The present data suggest that apigenin should be considered for use in the treatment of osteoporosis. PMID- 18508087 TI - 'Our shared responsibility': participation in ecological projects as a means of empowering communities to contribute to coastal management processes. AB - The objective of this project was to empower people of Blyth Valley in Northumberland (northeast England) to contribute to the management of their coastal heritage. A group of volunteers, the Links Conservation Group, working in collaboration with the environmental managers (Blyth Valley Borough Council), scientists, local specialists and other volunteers, surveyed the biodiversity, social, cultural and industrial history and profiles of the dunes, as well as their perceived value to local residents. The Group used this information to develop management plans for the dunes. The recommendations have contributed to planning processes at three levels: (i) national coastal management plans; (ii) regional plans; and (iii) plans for further action at the local community level. PMID- 18508088 TI - Ecological status assessment in the lower Eo estuary (Spain). The challenge of habitat heterogeneity integration: a benthic perspective. AB - The implementation of the European Water Framework Directive (WFD) requires evaluation of the ecological status (ES) of benthic communities in coastal and transitional water systems, and the intercalibration of assessment methodologies therefore becomes a research challenge. Our aim was to test the suitability of applying the M-AMBI index to assess the status of the Eo estuary (northern coast of Spain). Our results showed that M-AMBI was influenced by the natural variability of benthic communities, and presented an apparent dependence on habitat characteristics. Consequently, the definition of homogeneous areas in transitional water systems should be based on the salinity gradient combined with other factors. To achieve an accurate ES assessment, habitat-specific reference conditions should then be defined prior to the application of M-AMBI; this necessitates dividing an estuary into several sections, which may be classified as different ESs. From this perspective, a novel approach to integrate habitat heterogeneity in a global ES assessment was tested. PMID- 18508089 TI - Is there any seasonal variation in marine nematodes within the sediments of the intertidal zone? AB - The sediment parameters and nematode assemblages in the intertidal zone of the Hichirippu shallow lagoon, Hokkaido, Japan, were investigated. The objectives of this study were to observe the seasonal variation in the nematodes in the sediment, and to investigate the relationships between the nematodes and environmental factors. Samples were collected bi-monthly from five stations on the tidal flat from April 2003 to February 2004. It was found that the sediment parameters (Chl a concentration, AVS, TOC and TN contents) varied throughout the 10-month study. Fifty-four species of nematodes were found in the study area. The density and biomass of the nematodes varied in accordance with the sediment temperature during the sampling period. In this study, there was a seasonal variation in the nematode assemblage found in the intertidal zone of this shallow lagoon. The important factors affecting this variation were sediment temperature, and food competition among the nematodes themselves. The seasonal variation of the nematode also showed a relationship with the Chl a concentration in the sediment during the sampling period. PMID- 18508090 TI - Presence and origin of polycyclic aromatic hydrocarbon in sediments of nine coastal lagoons in central Vietnam. PMID- 18508091 TI - Environmental hypoxia as a metabolic constraint on fish: the case of Atlantic cod, Gadus morhua. AB - Hypoxia is known to provoke a wide range of effects on aquatic animals. Here we use laboratory and field data on Atlantic cod, Gadus morhua, to illustrate that many of these responses can be explained within the metabolic scope (MS) framework, i.e. taking into account the directive and limiting effects of dissolved oxygen (DO) on the ability of animals to acquire energy for growth and activity. A MS model for cod shows that scope for activity (swimming, feeding, etc.) is proportional to DO and becomes nil, jeopardising survival, when DO is < approximately 20% air saturation. Laboratory studies have confirmed this lethal threshold and demonstrated that growth and food ingestion were significantly reduced below 70% sat. This loss of appetite has been linked to a reduction of the peak value and an increase in duration of postprandial metabolism, in agreement with the MS model. Dwindling MS during hypoxia imposes an upper limit to swimming performance. Cod may also opt to reduce spontaneous swimming activity to spare oxygen for other activities such as digestion. In the Kattegat, the Baltic Sea, and the Gulf of St. Lawrence, eastern Canada, cod completely avoid waters where their MS is near zero. Furthermore, cod density increases exponentially with DO up to approximately 70% sat in the Gulf of St. Lawrence. Although hypoxia results in other direct and indirect effects as well, the MS framework allows modelling of many of the responses to hypoxia for individual cod that ought to be reflected at the population and community levels. The MS framework is also useful to compare species responses. We show that the impact of hypoxia on MS is similar, when expressed as a proportion of MS in normoxia, in cod, European sea bass (Dicentrarchus labrax), the common sole (Solea solea) and turbot (Psetta maxima). Data are required for other species to evaluate how general these findings are. PMID- 18508092 TI - Subtropical meiobenthic nematode communities in Victoria Harbour, Hong Kong. PMID- 18508093 TI - Radiation-induced bystander effects in vivo are sex specific. AB - Ionizing radiation (IR) effects span beyond the area of direct exposure and can be observed in neighboring and distant naive cells and organs. This phenomenon is termed a 'bystander effect'. IR effects in directly exposed tissue in vivo are epigenetically mediated and distinct in males and females. Yet, IR-induced bystander effects have never been explored in a sex-specificity domain. We used an in vivo mouse model, whereby the bystander effects are studied in spleen of male and female animals subjected to head exposure when the rest of the body is protected by a medical-grade lead shield. We analyzed the induction of DNA damage and alterations in global DNA methylation. Molecular parameters were correlated with cellular proliferation and apoptosis levels. The changes observed in bystander organs are compared to the changes in unexposed animals and animals exposed to predicted and measured scatter doses. We have found the selective induction of DNA damage levels, global DNA methylation, cell proliferation and apoptosis in exposed and bystander spleen tissue of male and female mice. Sex differences were significantly diminished in animals subjected to a surgical removal of gonads. These data constitute the first evidence of sex differences in radiation-induced bystander effects in mouse spleen in vivo. We show the role of sex hormones in spleen bystander responses and discuss implications of the observed changes. PMID- 18508094 TI - [Imaging for the management of cavernous sinus meningiomas]. AB - The treatment of cavernous sinus meningiomas remains controversial. However, at present, the general consensus is that only tumors with extracavernous extensions should be operated on and that resection is performed only on the extracavernous portions. Since these tumors are deep-seated and in the vicinity of vital and highly functional neurovascular structures, surgery remains difficult and dangerous. To achieve the gross total removal of the extracavernous extensions with minimal sequelae, the operative strategy must take into account the location of the tumor extension, not only in the basal cisterns but also into the skull base structures, the vascularization of the tumor with its feeders, as well as the venous drainage pathways. These anatomical and surgical data are collected from preoperative imaging, which includes CT-scan with bony windows and consecutive fine sections, MRI, angio-MR, and DSA angiography performed with selective arteriography including late venous phases. The operative plan must define the type of craniotomy and complementary osteotomy, the usefulness of an anterior clinoidectomy with skeletonization of the optic nerve, and determine whether extensive exposure of the cerebral middle fossa is necessary to ensure substantial devascularization of the tumor and proximal control of the internal carotid artery at its intrapetrosal portion. In addition, the possible need for an extra-intracranial bypass prior to tumor removal should be evaluated. The high number of neuroimaging investigations is the guarantee of surgical accuracy and safety. PMID- 18508095 TI - Calcium release via activation of presynaptic IP3 receptors contributes to kainate-induced IPSC facilitation in rat neocortex. AB - We examined the mechanisms of kainate (KA) induced modulation of GABA release in rat prefrontal cortex. Pharmacologically isolated IPSCs were recorded from visually identified layer II/III pyramidal cells using whole-cell patch clamp techniques. KA produced an increase in evoked IPSC amplitude at low nanomolar concentrations (100-500 nM). The frequency but not the amplitude of miniature (m) IPSCs was also increased. The GluR5 subunit selective agonist (RS)-2-amino-3-(3 hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA) caused an increase in mIPSC frequency whereas (3S,4aR,6S,8aR)-6-(4-carboxyphenyl)methyl 1,2,3,4,4a,5,6,7,8,8a-decahydroisoquinoline-3-carboxylic acid (LY382884), a selective GluR5 subunit antagonist, inhibited this facilitation. Philanthotoxin 433 (PhTx) blocked the effect of KA, indicating involvement of Ca(2+)-permeable GluR5 receptors. No IPSC facilitation was seen when Ca(2+) was omitted from the bathing solution. Facilitation was observed when slices were preincubated in ruthenium red or high concentrations of ryanodine, but was inhibited with application of thapsigargin. The IP3 receptor (IP3R) antagonists diphenylboric acid 2-amino-ethyl ester (2-APB) (15 microM) and Xestospongin C (XeC) blocked IPSC facilitation. These results show that activation of KA receptors (KARs) on GABAergic nerve terminals results is linked to intracellular Ca(2+) release via activation of IP3, but not ryanodine, receptors. This represents a new mechanism of presynaptic modulation whereby Ca(2+) entry through Ca(2+)-permeable GluR5 subunit containing KARs activates IP3Rs receptors leading to an increase in GABA release. PMID- 18508096 TI - Implication of neuropeptide-Y Y2 receptors in the effects of immune stress on emotional, locomotor and social behavior of mice. AB - Neuropeptide Y (NPY) is involved in the regulation of emotional behavior, and there is indirect evidence for a role of NPY in the cerebral responses to peripheral immune challenge. Since the NPY receptors involved in these reactions are not known, we investigated the effect of Escherichia coli lipopolysaccharide (LPS) on emotional, locomotor and social behavior, body temperature and circulating corticosterone in female Y2 (Y2-/-) and Y4 (Y4-/-) receptor knockout mice. LPS (0.1mg/kg injected IP 2.5h before testing) increased rectal temperature in control and Y4-/- mice to a larger degree than in Y2-/- animals. Both Y2-/- and Y4-/- mice exhibited reduced anxiety-related and depression-like behavior in the open field, elevated plus-maze and tail suspension test, respectively. While depression-like behavior was not changed by LPS, anxiety-related behavior was enhanced by LPS in Y2-/-, but not control and Y4-/- animals. Y2-/- mice were also particularly susceptible to the effect of LPS to attenuate locomotor behavior and social interaction with another mouse. The corticosterone response to LPS was blunted in Y2-/- mice which presented elevated levels of circulating corticosterone following vehicle treatment. These data show that Y2-/- mice are particularly sensitive to the effects of LPS-evoked immune stress to attenuate locomotion and social interaction and to increase anxiety-like behavior, while the LPS-induced rise of temperature and circulating corticosterone is suppressed by Y2 receptor knockout. Our observations attest to an important role of endogenous NPY acting via Y2 receptors in the cerebral response to peripheral immune challenge. PMID- 18508098 TI - Effects of neonatal oxytocin treatment on aggression and neural activities in mandarin voles. AB - Neonatal manipulation of oxytocin (OT) has long-term effects on behavior and physiology. Here we test the hypothesis that neonatal OT treatment can affect the subsequent expression of intrasexual aggression partly by reprogramming the neural activities of relevant brain regions. To test this hypothesis, mandarin voles (Lasiopodomys mandarinus) received OT or isotonic saline treatment within 24 h of birth. At about 75 days of age, aggressive behaviors and Fos expression in different brain regions were tested. The results indicate that the (1) level of intrasexual aggression was higher and other social contact was lower in SAL treated sexually naive males than in females and; (2) OT-treated females showed a greater increase in aggressive behaviors and Fos expression only after exposure to a male than SAL-treated females, but there were no significant changes in aggressive behaviors in males. These results demonstrate a sexual difference in aggression, and that neonatal exposure to OT may increase aggression in female mandarin voles. These effects may be based on changes in neural activities of relevant brain regions including the bed nucleus of the stria terminalis (BNST), lateral septal nucleus (LS), medial preoptic area (MPOA), the paraventricular nucleus of the hypothalamus (PVN), supraoptic nucleus (SON), mediodorsal thalamic nucleus (MD), ventromedial nucleus of hypothalamic (VMH), the medial amygdala (MeA) and central amygdala (CeA). PMID- 18508097 TI - Leptin resistance and the response to positive energy balance. AB - Animals readily reduce food intake and normalize body weight following a period of involuntary overfeeding, suggesting that regulatory systems are engaged to defend against excess weight gain. However, these data exist in the background of an ongoing obesity epidemic, where the ready availability of palatable, energy dense foods often leads to obesity. Currently we know very little about the mechanisms underlying the normalization of body weight following involuntary overfeeding, nor do we fully understand why select individuals successfully remain lean despite living in an obesigenic environment. Recent progress in the study of leptin signaling indicates that manipulations which enhance leptin sensitivity reduce food intake and attenuate diet-induced obesity, while reductions in leptin signaling predispose to obesity. While it remains unclear whether a failure or insufficiency in the weight regulatory system contributes to obesity, this work highlights the importance of this system for the regulation of body weight and its potential value for the treatment of obesity. Nonetheless, it is necessary to more clearly identify those mechanisms that protect lean individuals from weight gain and mediate the normalization of body weight that follows involuntary overfeeding, because it is only with this knowledge that we can clearly determine whether obesity is dependent on, or independent of, a failure in the weight regulatory system. PMID- 18508099 TI - Cognitive dietary restraint: impact on bone, menstrual and metabolic status in young women. AB - We compared bone mineral density (BMD) and content (BMC), menstrual and metabolic status between physically active women with 1) high cognitive dietary restraint (High-CDR) (score > or = 9, n=38) and Normal-CDR (score<9, n=46) and 2) across quartiles of CDR scores. Eighty-four physically active (500+/-35 min wk(-1)) premenopausal women participated and were categorized according to their CDR score. Primary outcomes included, BMD, BMC, menstrual status, estrone-3 glucuronide (E1G) and pregnanediol-3-glucuronide (PdG) area under the curve (AUC). Secondary outcomes included resting energy expenditure (REE), total triiodothyronine, and ghrelin. Measures of body mass (59.2+/-1.1 vs. 58.5+/-1.0 kg) and percent body fat (24.7+/-1.2 vs. 23.7+/-0.7%) were similar between women with Normal-CDR and High-CDR, however the High-CDR group had lower total body (1.140+/-0.011 vs. 1.179+/-0.010 g cm(-2); p=0.015) and lumbar spine (1.114+/ 0.019 vs. 1.223+/-0.022 g cm(-2); p=0.001) BMD. The prevalence of oligo amenorrhea was higher in the High-CDR group and became increasingly greater across the CDR quartiles. There were no differences in metabolic characteristics between the High-CDR and Normal-CDR groups, however REE and the ratio of measured to predicted REE were lower in the fourth quartile (CDR scores > or = 13) compared to the second and third quartiles. Our results provide evidence that high CDR scores are associated with reduced lumbar spine and total body BMD in physically active premenopausal women. A greater frequency of menstrual disturbances in women with higher CDR scores likely played a role in the reduced total body and lumbar spine BMD. PMID- 18508101 TI - Transmission-virulence trade-offs in vector-borne diseases. AB - Though it is commonly supposed that there is a trade-off between virulence and transmission, there is little data and little insight into what it should look like. Here, we consider the specific case of vector-borne parasites (inspired by human malaria) and analyse an embedded model to understand how specific life cycle aspects may affect this trade-off. First, we find that, for such parasites, the transmission function may have an S-shape. Second, we find that the trade-off obtained for vector-borne parasites is less sensitive to parameter variations than the trade-off obtained for directly transmitted parasites. Third, we find that other parasite traits, such as the conversion from replicative to infective stages, could have important epidemiological implications. Finally, we compare the effect of treatments targeting either the asexual or the sexual parasite life stage. PMID- 18508100 TI - CNS GLP-1 regulation of peripheral glucose homeostasis. AB - Current models hold that peripheral and CNS GLP-1 signaling operate as distinct systems whereby CNS GLP-1 regulates food intake and circulating GLP-1 regulates glucose homeostasis. There is accumulating evidence that the arcuate nucleus, an area of the CNS that regulates energy homeostasis, responds to hormones and nutrients to regulate glucose homeostasis as well. Recent data suggest that GLP-1 may be another signal acting on the arcuate to regulate glucose homeostasis challenging the conventional model of GLP-1 physiology. This review discusses the peripheral and central GLP-1 systems and presents a model whereby these systems are integrated in regulation of glucose homeostasis. PMID- 18508102 TI - Halothane-induced alterations in cellular structure and proliferation of A549 cells. AB - Genotoxicity, cytotoxicity or teratogenicity are among the well-known detrimental effects of the volatile anaesthetics. The aim of the present work was to study the structural changes, proliferative activity and the possibility of alveolar A549 cells to recover after in vitro exposure to halothane at 1.5 and 2.1mM concentrations. Our data indicated significant reduction of viability, suppression of mitotic activity more than 60%, and that these alterations were accompanied by disturbances of nuclear and nucleolar structures. The most prominent negative effect was the destruction of the lamellar bodies, the main storage organelles of pulmonary surfactant, substantial for the lung physiology. In conclusion, halothane applied at clinically relevant concentrations exerts genotoxic and cytotoxic effect on the alveolar cells in vitro, most likely as a consequence of stress-induced apoptosis, thus modulating the respiratory function. PMID- 18508103 TI - A structure-activity analysis of the variation in oxime efficacy against nerve agents. AB - A structure-activity analysis was used to evaluate the variation in oxime efficacy of 2-PAM, obidoxime, HI-6 and ICD585 against nerve agents. In vivo oxime protection and in vitro oxime reactivation were used as indicators of oxime efficacy against VX, sarin, VR and cyclosarin. Analysis of in vivo oxime protection was conducted with oxime protective ratios (PR) from guinea pigs receiving oxime and atropine therapy after sc administration of nerve agent. Analysis of in vitro reactivation was conducted with second-order rate contants (k(r2)) for oxime reactivation of agent-inhibited acetylcholinesterase (AChE) from guinea pig erythrocytes. In vivo oxime PR and in vitro k(r2) decreased as the volume of the alkylmethylphosphonate moiety of nerve agents increased from VX to cyclosarin. This effect was greater with 2-PAM and obidoxime (>14-fold decrease in PR) than with HI-6 and ICD585 (<3.7-fold decrease in PR). The decrease in oxime PR and k(r2) as the volume of the agent moiety conjugated to AChE increased was consistent with a steric hindrance mechanism. Linear regression of log (PR-1) against log (k(r2)[oxime dose]) produced two offset parallel regression lines that delineated a significant difference between the coupling of oxime reactivation and oxime protection for HI-6 and ICD585 compared to 2-PAM and obidoxime. HI-6 and ICD585 appeared to be 6.8-fold more effective than 2-PAM and obidoxime at coupling oxime reactivation to oxime protection, which suggested that the isonicotinamide group that is common to both of these oximes, but absent from 2-PAM and obidoxime, is important for oxime efficacy. PMID- 18508104 TI - The role of areas MT and MST in coding of visual motion underlying the execution of smooth pursuit. AB - What is the main purpose of visual motion processing? One very important aspect of motion processing is definitively the generation of smooth pursuit eye movements. These eye movements avoid motion blur of moving objects which would obstruct the analysis of the objects' visual details. However, these eye movements can only be executed if there is a moving target. So there is a very close and inseparable relationship between smooth pursuit and motion processing. The hub for visual motion processing is situated in the middle temporal (MT) and medial superior temporal (MST) area. Despite the undoubted importance of these areas for the generation of smooth pursuit or goal-directed behavior in general, it is important to keep in mind that motion processing in addition serves perceptual purposes such as object recognition, structure-from-motion detection, scene segmentation, self-motion estimation and depth perception. This review focuses at the beginning on pursuit-related activity recorded from MT and MST, subsequently extends the view to goal-directed hand movements, and finally addresses the possible contributions of these areas to motion perception. PMID- 18508105 TI - Effects of acute ethyl alcohol consumption on a psychophysical measure of lateral inhibition in human vision. AB - Acute consumption of ethyl alcohol affects a variety of visual functions. However, there have been few systematic attempts to investigate the neural mechanisms underlying these effects. Here, we employed the Westheimer paradigm to investigate the hypothesis that alcohol reduces lateral inhibition within human "perceptive fields", the psychophysical analogue of physiological receptive fields. Westheimer functions obtained under alcohol and no-alcohol conditions at photopic, mesopic, and scotopic levels of adaptation showed changes consistent with an alcohol-induced decrease in lateral inhibition. We conclude that this decrease in lateral inhibition may be responsible for some of the changes in visual perception that result from alcohol consumption. PMID- 18508106 TI - An overall isotherm for activated carbon adsorption of dissolved natural organic matter in water. AB - Design and analysis of activated carbon processes in water treatment often requires the adsorption isotherm for dissolved natural organic matter (NOM). Of the isotherm models available, the Summers and Roberts (SR) equation, capable of describing the adsorbent dose effect with the fewest parameters, has been successfully used to normalize NOM isotherm data. In this study, we show that the adsorbent dose in the SR equation can be eliminated as an intermediate variable and the initial concentration effect on NOM adsorption is then described explicitly. Comparing with the original SR equation, the derived isotherm equation is in a form more amenable to analysis. To ensure that the prediction is physically attainable, we introduced the limiting adsorption capacity by taking the adsorbent pore volume and size exclusion into consideration. Subsequently, we develop a simple relationship that can be used to determine the minimum adsorbent usage required for any desirable level of treatment. By comparing with extensive isotherm data previously published by Li et al. [2003a. Polydisperse adsorbability composition of several natural and synthetic organic matrices. J. Colloid Interface Sci. 265(2), 265-275], we demonstrated that the isotherm equation derived herein yields predictions that agree with the much more complicated fictive component-ideal adsorbed solution theory (IAST)-based model for NOM from different sources and over a range of initial concentrations. PMID- 18508107 TI - Arylstibonic acids: novel inhibitors and activators of human topoisomerase IB. AB - Human topoisomerase IB (hTopo) forms a covalent phosphotyrosyl linkage with the DNA backbone, and controls genomic DNA topology by relaxing DNA supercoils during the processes of DNA replication, transcription, chromosome condensation and decondensation. The essential role of hTopo in these processes has made it a preeminent anticancer drug target. We have screened a small library of arylstibonic acids for their effects on plasmid supercoil relaxation catalyzed by hTopo. Despite the similar structures of the library compounds, some compounds were found to be effective competitive inhibitors, and others, nonessential activators. Some arylstibonic acids show selectivity in their action against hTopo and the related enzyme from poxvirus (vTopo). Structure-activity relationships and structural modeling suggest that competitive inhibition may result from positioning of the negatively charged stibonic acid and carboxylate groups of the inhibitors into DNA phosphate binding pockets on hTopo. The hTopo activators act by a surprising allosteric mechanism without interfering with DNA binding or binding of the widely used hTopo poison camptothecin. Arylstibonic acid competitive inhibitors may become useful small molecules for elucidating the cellular functions of hTopo. PMID- 18508108 TI - Quantitative structure-property relationships for octanol-water partition coefficients of polybrominated diphenyl ethers. AB - Theoretical molecular descriptors were tested against logK(OW) values for polybrominated diphenyl ethers (PBDEs) using the Partial Least-Squares Regression method which can be used to analyze data with many variables and few observations. A quantitative structure-property relationship (QSPR) model was successfully developed with a high cross-validated value (Q(cum)(2)) of 0.961, indicating a good predictive ability and stability of the model. The predictive power of the QSPR model was further cross-validated. The values of logK(OW) for PBDEs are mainly governed by molecular surface area, energy of the lowest unoccupied molecular orbital and the net atomic charges on the oxygen atom. All these descriptors have been discussed to interpret the partitioning mechanism of PBDE chemicals. The bulk property of the molecules represented by molecular surface area is the leading factor, and K(OW) values increase with the increase of molecular surface area. Higher energy of the lowest unoccupied molecular orbital and higher net atomic charge on the oxygen atom of PBDEs result in smaller K(OW). The energy of the lowest unoccupied molecular orbital and the net atomic charge on PBDEs oxygen also play important roles in affecting the partition of PBDEs between octanol and water by influencing the interactions between PBDEs and solvent molecules. PMID- 18508109 TI - Nuclear factor-kappaB activation is associated with somatic and germ line RET mutations in medullary thyroid carcinoma. AB - The nuclear factor-kappaB (NF-kappaB) family of transcription factors regulates a wide variety of cellular processes including cell growth, differentiation, and apoptosis. NF-kappaB has been shown to be activated through several signaling pathways that involve growth factor receptors. The aim of the study was to assess the immunohistochemical expression of members of the NF-kappaB family and the putative targets of NF-kappaB in a series of medullary thyroid carcinomas (MTCs), in correlation with RET mutational status. A tissue microarray was constructed from paraffin-embedded blocks of 48 MTCs (13 familial, 35 sporadic) previously evaluated for germ line and somatic RET mutations. Immunohistochemical evaluation included members of the NF-kappaB (p50, p65, p52, c-Rel, RelB) family, as well as putative targets of NF-kappaB such as Flip, Bcl-xL, and cyclin D1. Nuclear immunostaining for members of NF-kappaB was frequent in MTCs (p50, 19%; p65, 68%; p52, 86.6%; c-Rel, 75%; RelB, 36%). MTCs with germ line or somatic RET mutations (29 cases) showed NF-kappaB nuclear translocation (particularly of p65, P = .035) more frequently than MTCs without RET mutations (19 cases). Immunostaining for putative targets of NF-kappaB showed a significant statistical association between p65 and Bcl-xL (P = .024). In addition, Bcl-xL expression was statistically higher in the tumors with exon 16 RET mutation in comparison with those with exon 10 and 11 RET mutations or wild-type RET (P = .002). Moreover, the significance of RETsignaling in NF-kappaB activation was evaluated in the RET mutated TT cell line. TT cells were infected with lentiviruses carrying short hairpin RNA to knock down RET expression, and NF-kappaB activity was assessed by luciferase reporter assays. Silencing of RET in the TT cell line produced a significant decrease in NF-kappaB activation and reduction in ERK1/2. The results suggest that the NF-kappaB is frequently activated in MTCs. The results also support the hypothesis that RET activation by somatic or germ line mutations may be responsible for NF-kappaB activation in MTCs. PMID- 18508110 TI - Hamartomas of mature cardiac myocytes: report of 7 new cases and review of literature. AB - Only 8 cases of hamartomas of mature cardiac myocytes have been reported. The aim of the study was to describe 7 new cases and provide clinicopathologic correlation. Our anatomical pathology database was searched for all cases of cardiac hamartoma, of which 7 represented mature myocyte type. Medical records were reviewed for clinical information, and microscopic slides were evaluated for extent of characteristics. Five males and 2 females ranged in age from 6 months to 74 years (mean, 23 years). There were 11 ventricular hamartomas (8 left free wall, 2 right free wall, 1 septum). Death in 3 infants was unrelated to incidental hamartomas discovered at autopsy. A 10- and 16-year-old were asymptomatic but had abnormal electrocardiogram (ECG) results, which led to detection of cardiac masses by imaging studies. Two adult males had only mild coronary disease angiographically. The 57-year-old, who died suddenly, had a 7 year history of abnormal ECG results. The 74-year-old, who died after aortic surgery, had a 3-year history of chest discomfort. Their hamartomas were identified at autopsy and contributed to sudden death in 1. Microscopically, all hamartomas were involved by myocyte hypertrophy and disarray, without inflammation or calcification. Myocyte vacuolization and venular dilatation occurred only in the pediatric cases, and interstitial adipose tissue only affected 1 adult. In conclusion, hamartomas of mature cardiac myocytes may be detected at any age. They primarily affect males, arise predominantly in the left ventricle, are asymptomatic, may have nonspecific ECG findings, and rarely may be associated with sudden death. Microscopic findings in infants differ from older patients. PMID- 18508111 TI - The exposure of temple workers to polycyclic aromatic hydrocarbons. AB - Five temples, each a different size, were selected for this study. Two of the temples were located in Taichung City (in central Taiwan), and three were in Tainan City (in Southern Taiwan). Aerosols smaller than 10 microm aerodynamic diameter (PM(10)) were collected by using personal collection samplers during pilgrim days (the first and fifteenth day of each lunar month) and normal days (all other days). Regression analysis showed that about 1.6 microg/m(3) of PM(10) contributed to the workers' exposure in the temples for each joss stick increase in the censer. The concentrations of total polycyclic aromatic hydrocarbons (t PAHs) and carcinogenic PAHs (car-PAHs) on pilgrim days were higher than those on normal days. Mean concentrations of urinary 1-hydroxypyrene (1-OHP) in the pre shift and post-shift workers of the five temples on normal days were 1.20 and 1.61 microg/g creatinine, respectively. Furthermore, the post-shift concentrations of 1-OHP in the workers of temples on pilgrim days were all higher than those of the workers of corresponding temples on normal days. Significant relationships between the urinary concentrations of 1-OHP and the exposure concentrations of pyrene, total PAHs and BaP(eq) were found in the workers of the temples. Results of linear regression showed that the increase of unit concentration (1 ng/m(3)) of pyrene led to a 0.05 microg/g creatinine increase of urinary 1-OHP, while the increase of unit concentration (1 ng/m(3)) of BaP(eq) resulted in an increase of 0.03 microg/g creatinine of urinary 1-OHP. PMID- 18508112 TI - Damage effects induced by electrically generated negative air ions in Caenorhabditis elegans. AB - Electrically generated negative air ions (ENIs) have been widely used to improve indoor air quality. However, the effects of ENIs reported so far were inconsistent due to the variance in test systems, end points detected and the exposure methods. In this study, a simple model organism, the nematode Caenorhabditis elegans, was used as an in vivo system to assess the biological effects of continuous ENIs exposure. The worms were exposed to ENIs in a 10(5) 10(5) ions/cm(3) chamber and their development period, lifespan, brood size and germline cell apoptotisis were examined. The results showed that ENIs decreased the development period, shortened the lifespan, increased the germline cell apoptosis and reduced the brood size, suggesting that persistent ENIs exposure might induce damage in C. elegans. To further scrutinize the mechanisms underlying these damage effects, a reactive oxygen species (ROS) sensitive C. elegans, mev-1(kn1) mutant, and sod-3::gfp transgenic strains were used. The results showed that the persistent ENIs exposure significantly shortened the lifespan of mev-1(kn1) mutant compared to the wild type. Moreover, levels of SOD 3 were increased in an exposure time-dependent manner. Treatment with either DMSO or l-ascorbic acid, effective ROS scavengers, could rescue the upregulation of germline cell apoptosis and SOD-3 level induced by ENIs exposure, indicating that ROS may be involved in ENIs exposure-induced damaging effects. PMID- 18508113 TI - The Water Framework Directive: total environment or political compromise? AB - The European Water Framework Directive (2000/60/EC) is potentially ground breaking legislation. It seeks to bring about improvement of aquatic habitats in Europe to 'good ecological status', defined as slightly different from 'high ecological status', with no or minimal human impact. The characteristics of pristine ecological status include nutrient parsimony, a defined characteristic structure of the system (including geomorphological structure and hydrology, biological and food web structure) and the connectivity and extent of the system that are essential for resilience to change. This modern ecological understanding is being ignored by government agencies charged with enacting the Directive. Schemes are being devised that measure secondary characteristics of habitats using approaches drawn from traditional water quality management. Typologies, indicated by the Directive to give a geographical basis within which to determine ecological status, are also being corrupted with different typologies used for different determinands. The ecological reality of reasonably distinctive, integrated systems (an erosive upland river versus a floodplain system, for example) is being avoided. Emphasis is being placed on precision of measurement of specific determinands rather than accuracy in what is being measured and proposed schemes are complex and expensive when accurate assessment could be carried out much more cheaply. Many are also likely to become redundant as effects of climate change take hold. The current approach will lead to some improvement in water quality but not to the fundamental change in ecological quality intended by the Directive and has partly been encouraged by lack of definition and contradictions within the Directive itself. Documented details currently available from the UK agencies are used to illustrate how the intentions of the Directive are being undermined for ostensibly political convenience through processes of redefinition and limitation of characteristics measured. There appears to be a parallel concern among official and non governmental European bodies. PMID- 18508114 TI - Heterogeneous glycosylation patterns of human PAI-1 may reveal its cellular origin. AB - The main inhibitor of intravascular fibrinolysis is plasminogen activator inhibitor 1 (PAI-1) which binds to and irreversibly inhibits tissue plasminogen activator (tPA). PAI-1 is present in blood, both in platelets and in plasma, and PAI-1 levels are associated with risk of atherothrombosis. Several tissues express PAI-1 but the source of plasma PAI-1 is not known. We recently found that platelets can de novo synthesize PAI-1 and the amount synthesized in vitro in 24 hours is 35-fold higher than required to maintain normal plasma levels. Recombinant human PAI-1 expressed in different cell types or secreted naturally by human cell lines, exhibit heterogeneous glycosylation patterns. The aim of this study was to investigate the hypothesis that platelets might be the source of plasma PAI-1 and that the cellular source of PAI-1 can be determined by its tissue-specific glycosylation pattern. PAI-1 was isolated from platelets, macrophages, endothelial cells, adipose tissue, as well as plasma from lean and obese subjects. The glycosylation was analyzed by nanoLC-MS/MS. PAI-1 isolated from cell lysates and conditioned media from macrophages, endothelial cells, and adipose tissue expressed heterogeneous glycosylation patterns. By contrast, no glycans were detected on PAI-1 isolated from plasma or platelets from healthy lean individuals. Hence, our data suggest that platelets may be the main source of plasma PAI-1 in lean individuals. Interestingly, plasma PAI-1 from obese subjects had a glycan composition similar to that of adipose tissue suggesting that obese subjects with elevated PAI-1 levels may have a major contribution from other tissues. PMID- 18508115 TI - Frequency of maternal licking and grooming correlates negatively with vulnerability to cocaine and alcohol use in rats. AB - Because licking and grooming behavior of dams with pups can influence some behaviors of pups when they are adults, we tested if licking and grooming scores in a maternal separation protocol correlated with cocaine or ethanol self administration in the pups as adults. The protocol produced litters that were separated from dams for 0 (MS0), 15 (MS15) or 180 (MS180) min, and a nonhandled (NH) group as well. Self-administration of both drugs as shown in earlier studies was lowest in the MS15 group, highest in the NH group and intermediate in the other groups. Licking and grooming scores correlated negatively with drug intake and suggests that maternal care of pups can influence drug use in pups when they are adults. PMID- 18508116 TI - Prefrontal cortex-nucleus accumbens interaction: in vivo modulation by dopamine and glutamate in the prefrontal cortex. AB - Previous experimental studies have shown that the prefrontal cortex (PFC) regulates the activity of the nucleus accumbens (NAc), and in particular the release of dopamine in this area of the brain. In the present report we review recent microinjections/microdialysis studies from our laboratory on the effects of stimulation/blockade of dopamine and glutamate receptors in the PFC that modulate dopamine, and also acetylcholine release in the NAc. Stimulation of prefrontal D2 dopamine receptors, but not group I mGlu glutamate receptors, reduces the release of dopamine and acetylcholine in the NAc and spontaneous motor activity. This inhibitory role of prefrontal D2 receptors is not changed by acute systemic injections of the NMDA antagonist phencyclidine. On the other hand, the blockade of NMDA receptors in the PFC increases the release of dopamine and acetylcholine in the NAc as well as motor activity which suggests that the hypofunction of prefrontal NMDA receptors is able to produce the neurochemical and behavioural changes associated with a dysfunction of the corticolimbic circuit. We suggest here that dopamine and glutamate receptors are, in part, segregated in specific cellular circuits in the PFC. Thus, the stimulation/blockade of these receptors would have a different net impact on PFC output projections to regulate dopamine and acetylcholine release in the NAc and in guided behaviour. Finally, it is speculated that environmental enrichment might produce plastic changes that modify the functional interaction between the PFC and the NAc in both physiological and pathological conditions. PMID- 18508117 TI - Echo-Doppler ultrasonographic assessment of resistance and velocity of blood flow in the ductus venosus throughout gestation in fetal lambs. AB - It has been postulated that during fetal hypoxia, the blood flow shunted through the ductus venosus increases and may account for upto 70% of the total umbilical flow. The objectives of the present study were to use ultrasonography to determine the velocities and waveform indices of blood flow in the ductus venosus in the fetal lamb. The ductus venosus of 15 lamb fetuses was evaluated weekly from Days 45 to 143 of gestation (Day 0=day of AI). The Doppler indices measured were: S/D, the systole/diastole ratio; RI, the resistance index; and PI, the pulsatility index. The velocity waveforms studied were: SV, the peak velocity during ventricular systole; D, the peak velocity during ventricular diastole; aV, the lowest forward velocity during atrial contraction; and TAMV, the time averaged maximum velocity. Doppler indices from Days 52 to 143 were highly correlated: S/D versus PI (r=0.96, P<0.0001), and versus RI (r=0.93, P<0.0001); and PI versus RI (r=0.97, P<0.0001). Velocity indices were also positively correlated: velocity SV versus D (r=0.87, P<0.0001), versus aV (r=0.79, P<0.05), and versus TAMV (r=0.98, P<0.0001); D versus aV (r=0.88, P<0.05), and versus TAMV (r=0.87, P<0.05); and aV versus TAMV (r=0.82, P<0.05). Doppler indices were negatively correlated with SV (r=-0.22, P<0.03); D (r=-0.37, P<0.0001); TAMV (r= 0.32, P<0.05) and with aV (r=-0.67, P<0.05). The mean value of each Doppler index decreased 40% from Days 52 to 73 (e.g., PI from 0.82+/-0.08 to 0.51+/-0.10; P<0.05), with no significant changes thereafter. Mean (+/-S.E.M.) values of velocity indices SV, D, aV, and TAMV rose twofold from Days 60 to 115 of gestation (e.g., SV from 54.4+/-8.8cm/s to 104.9+/-19.7 and aV from 24.8+/-6.9 to 54.9+/-5.9; P<0.05). In conclusion, Doppler ultrasonography facilitated assessment of the blood flow pattern in the ductus venosus in lamb fetuses between Days 52 and 143. PMID- 18508118 TI - Development of photocrosslinked sialic acid containing polymers for use in Abeta toxicity attenuation. AB - beta-Amyloid peptide (Abeta), the primary protein component in senile plaques associated with Alzheimer's disease (AD), has been implicated in neurotoxicity associated with AD. Previous studies have shown that the Abeta-neuronal membrane interaction plays a crucial role in Abeta toxicity. More specifically, it is thought that Abeta interacts with ganglioside rich and sialic acid rich regions of cell surfaces. In light of such evidence, we have hypothesized that the Abeta membrane sialic acid interaction could be inhibited through use of a biomimic multivalent sialic acid compound that would compete with the cell surface for Abeta binding. To explore this hypothesis, we synthesized a series of photocrosslinked sialic acid containing oligosaccharides and tested their ability to bind Abeta and attenuate Abeta toxicity in cell culture assays. We show that a polymer prepared via the photocrosslinking of disialyllacto-N-tetraose (DSLNT) was able to attenuate Abeta toxicity at low micromolar concentrations without adversely affecting the cell viability. Polymers prepared from mono-sialyl oligosaccharides were less effective at Abeta toxicity attenuation. These results demonstrate the feasibility of using photocrosslinked sialyl-oligosaccharides for prevention of Abeta toxicity in vitro and may provide insight into the design of new materials for use in attenuation of Abeta toxicity associated with AD. PMID- 18508119 TI - Dissociation of the anxiolytic-like effects of Avpr1a and Avpr1b receptor antagonists in the dorsal and ventral hippocampus. AB - Arginine-vasopressin (AVP) is synthesized and released centrally in several brain structures. AVP is thought to mediate anxiety-related behavior through two central receptor subtypes, Avpr1a and Avpr1b. Although these AVP receptor subtypes are expressed in several brain regions, including the hippocampus, little is known about their explicit role in unconditioned fear or anxiety. This experiment assessed the anxiety-related effects of a selective Avpr1a antagonist ([beta-Mercapto-beta,beta-cyclopentamethylenepropionyl1, O-me-Tyr2, Arg8]-AVP) and a selective Avpr1b antagonist ((2S,4R)-1-[5-chloro-1-[(2,4 dimethoxyphenyl)sulfonyl]-3-(2-methoxy-phenyl)-2-oxo-2,3-dihydro-1H-indol-3-yl]-4 hydroxy-N,N-dimethyl-2-pyrrolidine carboxamide; SSR 149415) microinfused into either the dorsal or ventral sub-regions of the rat hippocampus. Avpr1a antagonism in the ventral, but not the dorsal hippocampus reduced rats' anxiety like behavior in the elevated plus-maze test. Conversely, Avpr1b antagonism in the dorsal, but not the ventral, hippocampus reduced anxiety in the plus-maze test. Neither antagonist reduced anxiety-like behavior in the shock-probe burying test. Overall, the results show that both receptor subtypes of AVP are involved in anxiety-related responses, but their specific contributions depend on three variables: (1) the anxiety-related response (shock-probe avoidance versus open arm avoidance), (2) the receptor subtype antagonized (Avpr1a versus Avpr1b), and (3) the area of hippocampus (dorsal versus ventral) into which these antagonists are infused. These dissociations suggest that different fear responses are under the control of specific AVP receptor systems within discrete parts of the hippocampus. PMID- 18508120 TI - Central nervous system involvement in adult acute lymphoblastic leukemia at diagnosis and/or at first relapse: results from the GET-LALA group. AB - Outcome of adult acute lymphoblastic leukemia (ALL) with central nervous system (CNS) involvement is not clearly defined. We studied 104 patients presenting with CNS involvement at diagnosis among 1493 patients (7%) included into the LALA trials, and 109 patients presenting CNS disease at the time of first relapse among the 709 relapsing patients (15%). Eighty-seven patients (84%) with CNS leukemia at diagnosis achieved complete remission (CR). Fifty-three patients underwent stem cell transplantation (SCT): 25 allogeneic SCT, 28 autologous SCT, while 34 continued with chemotherapy alone. Seven-year overall survival (OS) and disease-free survival (DFS) were 34% and 35%, respectively. There were no significant differences in terms of CR, OS and DFS among patients with CNS involvement at diagnosis and those without CNS disease. There were also no differences among the two groups regarding T lineage ALL, B lineage ALL, and among those who underwent SCT. After a first relapse, 38 patients with CNS recurrence (35%) achieved a second CR. The median OS was 6.3 months. Outcome was similar to that of relapsing patients without CNS disease. CNS leukemia in adult ALL is uncommon at diagnosis as well as at the time of first relapse. With intensification therapy, patients with CNS leukemia at diagnosis have a similar outcome than those who did not present with CNS involvement. CNS leukemia at first relapse remains of similar poor prognosis than all other adult ALL in first relapse. PMID- 18508121 TI - Comparing the mutagenicity of toxaphene after aging in anoxic soils and accumulating in fish. AB - A test program was conducted to evaluate the mutagenicity of toxaphene residuals extracted from aged soils and from fish collected in creeks near a toxaphene contaminated site. The ultimate objective was to determine if the residual toxaphene congeners were more or less mutagenic than those in technical-grade toxaphene. The study showed that the mutagenicity of the bioaccumulated toxaphene congeners in fish, expressed as colony revertants per microg of residual toxaphene, was no greater than that of technical-grade toxaphene. The mutagenic impact of the toxaphene residuals in aged soil statistically was less than that for technical-grade toxaphene. Two specific congeners, a hexachlorobornane (labeled Hx-Sd) and a heptachlorobornane (labeled Hp-Sd), were found to accumulate over time in both soil and fish extracts, but did not show increased mutagenic impacts relative to that produced by technical-grade toxaphene. PMID- 18508122 TI - Predictive validity of the Structured Assessment of Violence Risk in Youth (SAVRY) during residential treatment. AB - This prospective study examines the predictive validity of the Dutch version of the Structured Assessment of Violence Risk in Youth (SAVRY) by examining relationships between SAVRY scores and various types of disruptive behavior during residential treatment. The SAVRY, a risk assessment instrument, was coded for 66 male adolescents on the basis of file information and interviews. The adolescents were referred to Rentray, a juvenile correctional and treatment facility, by the Dutch juvenile courts because of severe behavioral problems or serious offenses. Institutional infractions were retrieved from incident registration files, which included acts of physical violence, verbal threat, verbal abuse, and violation of institutional rules. The interrater reliability of the SAVRY scores was good. The predictive validity of the SAVRY for physical violence against persons was excellent (Risk Total: AUC=.80, r =.33; Summery Risk Rating: AUC =.86, r =.48). The SAVRY also had good predictive validity for violence against objects, verbal threats and violations of rules, but not for verbal abuse. Implications for assessment and management of violent behavior among adolescents in residential treatment are discussed. PMID- 18508123 TI - Occurrence and distribution of organochlorine pesticides - lindane, p,p'-DDT, and heptachlor epoxide - in surface water of China. AB - Persistent organochlorine pesticides pollutants (OCPs) have been reported to occur at relatively high concentrations in some Chinese waters. In order to map the distribution of organochlorine pesticides in the surface water throughout China, samples were collected from over 600 sites in seven major river basins and three main internal rivers drainage areas during 2003 and 2004. The surface water samples were analyzed for the representative organochlorine pesticides contaminants including lindane (gamma-HCH), p,p'-DDT and heptachlor epoxide. In general, the most frequently detected compound was lindane, being detected in 83.9% of samples (mean=31.3 ng/l; range <0.17-860 ng/l), and the highest concentration was present in the Yellow River basin. p,p'-DDT was detected in 63.1% of the samples collected (mean=14.6 ng/l; range <0.14-368 ng/l) with the highest concentration present in the Huaihe River basin. Heptachlor epoxide was detected in only 9.3% of water samples (range <0.11-10 ng/l). Measured concentrations for the three compounds were low and rarely exceed the environment quality standard for surface water of China. Lindane was more frequently detected at much higher concentrations in the rivers of northern China compared with those of southern China. The sites with higher concentration of lindane and p,p'-DDT mainly occurred in the Yellow River and Huaihe River basins, so the results of this investigation indicate that the organochlorine pesticide contamination of Yellow River and Huaihe River basins should be of particular concern relative to the other basins. When compared with other regions of the world, it appears that the Chinese surface water is moderately polluted by lindane and p,p'-DDT. PMID- 18508124 TI - The crystal structure of halofantrine-ferriprotoporphyrin IX and the mechanism of action of arylmethanol antimalarials. AB - The crystal structure of the complex formed between the antimalarial drug halofantrine and ferriprotoporphyrin IX (Fe(III)PPIX) has been determined by single crystal X-ray diffraction. The structure shows that halofantrine coordinates to the Fe(III) center through its alcohol functionality in addition to pi-stacking of the phenanthrene ring over the porphyrin. The length of the Fe(III)-O bond is consistent with an alkoxide and not an alcohol coordinating group. The iron porphyrin is five coordinate and monomeric. Changes in the electronic spectrum of Fe(III)PPIX upon addition of halofantrine base in acetonitrile solution are almost identical to those observed upon addition of quinidine free base in the same solvent. This suggests homologous binding. Molecular mechanics modeling of Fe(III)PPIX complexes of quinidine, quinine, 9 epiquinine and 9-epiquinidine based on this homology suggests that the antimalarially active quinidine and quinine can readily adopt conformations that permit formation of an intramolecular salt bridge between the protonated quinuclidine tertiary amino group and unprotonated heme propionate group, while the inactive epimers 9-epiquinidine and 9-epiquinine have to adopt high energy conformations in order to accommodate such salt bridge formation. We propose that salt bridge formation may interrupt formation of the hemozoin precursor dimer formed during the heme detoxification pathway and so account for the strong activity of the two active isomers. PMID- 18508125 TI - TRP channel and cardiovascular disease. AB - The transient receptor potential (TRP) channel superfamily consists of 28 mammalian cation channels and is expressed in almost every tissue, including the heart and vasculature; most TRP channels are permeable to Ca(2+) and are prime molecular candidates for store-operated channels (SOCs), receptor-operated channels (ROCs), ligand-gated channels (LGCs) and stretch-activated channels (SACs). As these channels act as multifunctional cellular sensors and are involved in several fundamental cell functions such as contraction, proliferation and cell death, investigation of their roles in human disease is very important. This review presents an overview of current knowledge about the pathological role of TRP channels in cardiovascular diseases and highlights some TRP channels for which a role in the diseases can be anticipated. Evidences suggest that up regulation of TRPC channels is involved in the development of cardiac hypertrophy and heart failure; TRPM4 participates in some features of cardiac arrhythmias; increased expression of TRPC channels is associated with vascular remodeling and pulmonary hypertension; reduced expression or activity of TRPV4 impairs endothelium-dependent vasorelaxation; TRPC3/C4 and TRPM2 act as endothelial redox sensors; and TRPC1, -C4, -C6, -V4, and -M2, have been implicated in endothelial barrier dysfunction. Ultimately, TRP channels will become important novel pharmacological targets for the treatment of human cardiovascular diseases. PMID- 18508126 TI - Diacylglycerol acyltransferases: Potential roles as pharmacological targets. AB - Triglyceride (TG) synthesis occurs in many cell-types, but only the adipocyte is specialised for TG storage. The increased incidence of obesity and its attendant pathologies have increased interest in pharmacological strategies aimed at inhibition of triglyceride synthesis. In the liver this would also appear to offer the advantages of the prevention of steatosis and/or dyslipidaemia. The two major enzymes that have DGAT activity appear to have specialised functions, that are most evident in triglyceride-secreting tissues. The presence of triglyceride in non-adipose cells can lead to (through lipolysis), or be a marker for, undesirable complications such as insulin resistance, or can be indicative of simultaneously high capacities for triglyceride synthesis, lipolysis and oxidation of fatty acids as in highly aerobic, trained muscle. Consequently, inhibition of triglyceride synthesis may not be a straightforward strategy, either in terms of its achievement pharmacologically or in its anticipated outcomes. The metabolic complexities of triglyceride synthesis, with particular reference to the diacylglycerol acyltransferases (DGATs) are considered in this short review. PMID- 18508127 TI - Detection of neuronal spikes using an adaptive threshold based on the max-min spread sorting method. AB - Neuronal spike information can be used to correlate neuronal activity to various stimuli, to find target neural areas for deep brain stimulation, and to decode intended motor command for brain-machine interface. Typically, spike detection is performed based on the adaptive thresholds determined by running root-mean-square (RMS) value of the signal. Yet conventional detection methods are susceptible to threshold fluctuations caused by neuronal spike intensity. In the present study we propose a novel adaptive threshold based on the max-min spread sorting method. On the basis of microelectrode recording signals and simulated signals with Gaussian noises and colored noises, the novel method had the smallest threshold variations, and similar or better spike detection performance than either the RMS based method or other improved methods. Moreover, the detection method described in this paper uses the reduced features of raw signal to determine the threshold, thereby giving a simple data manipulation that is beneficial for reducing the computational load when dealing with very large amounts of data (as multi electrode recordings). PMID- 18508128 TI - Partial Granger causality--eliminating exogenous inputs and latent variables. AB - Attempts to identify causal interactions in multivariable biological time series (e.g., gene data, protein data, physiological data) can be undermined by the confounding influence of environmental (exogenous) inputs. Compounding this problem, we are commonly only able to record a subset of all related variables in a system. These recorded variables are likely to be influenced by unrecorded (latent) variables. To address this problem, we introduce a novel variant of a widely used statistical measure of causality--Granger causality--that is inspired by the definition of partial correlation. Our 'partial Granger causality' measure is extensively tested with toy models, both linear and nonlinear, and is applied to experimental data: in vivo multielectrode array (MEA) local field potentials (LFPs) recorded from the inferotemporal cortex of sheep. Our results demonstrate that partial Granger causality can reveal the underlying interactions among elements in a network in the presence of exogenous inputs and latent variables in many cases where the existing conditional Granger causality fails. PMID- 18508129 TI - Minor physical anomalies in affective disorders. A review of the literature. AB - BACKGROUND: The increased frequency of MPAs may be external markers of abnormal brain development in affective disorders. METHODS: A MEDLINE, psychInfo and Web of Science search was evaluated to collect all publications on the prevalence of minor physical anomalies in bipolar affective disorder and unipolar major depression. AIMS: As reports on the prevalence of MPAs in affective disorders were controversial, were based on highly different number of patients and were evaluated by the use of scales with different sensitivities, we considered as important to review the current state of knowledge and to recommend directions to further research. RESULTS: 14 publications on 12 studies were found after a careful literature search. 5 studies have dealt with the prevalence of MPAs in bipolar affective disorder, 3 have reported on examinations among patients with unipolar major depression, while 5 publications on 3 studies combined patients with bipolar affective disorder, schizoaffective disorder and unipolar major depression. 1 study was published on the prevalence of MPAs among mood disorders, without the differentiation of the data of patients with bipolar affective disorder and unipolar major depression. LIMITATIONS: Few studies with relatively small size were published, there is no data on the distinction between bipolar I and bipolar II disorders. CONCLUSION: The reviewed data suggest a higher probability of the role of an aberrant neurodevelopment in bipolar affective disorder and a smaller in unipolar major depression. PMID- 18508130 TI - Chemokine CXCL8 modulates GluR1 phosphorylation. AB - The chemokine interleukin 8/CXCL8 induces the phosphorylation of the GluR1 subunit of the AMPA-type glutamate receptor in neurons and transfected HEK cells, on both serine 845 (S845) and 831 (S831) residues. We previously described that CXCL8 receptor CXCR2 and GluR1 co-precipitate and that GluR1/CXCR2 co-expression both in HEK cells and neurons impairs CXCL8-induced cell migration. Here we show that replacement of S845 with Ala (A), but not with Glu (E), strongly reduces GluR1/CXCR2 interaction and abolishes the impairment of CXCL8-induced cell migration. Considered together our findings point to the phosphorylated state of S845GluR1 as a determinant of GluR1-CXCR2 physical coupling. PMID- 18508131 TI - Role of the chemokine receptor CX3CR1 in the mobilization of phagocytic retinal microglial cells. AB - We recently showed that subretinal CX3CR1-dependent microglial cell (MC) accumulation may lead to age-related macular degeneration. The fate of MC after engulfing retinal debris is poorly understood. Severe photoreceptor degeneration was observed 40days after exposure to bright light in CX3CR1-deficient but not control mice, and more MCs accumulated in the subretinal space of the former than the latter. To study the fate of subretinal MCs in CX3CR1 competent animals, we used a dystrophic rat model in which abundant subretinal MC accumulation is observed secondary to retinal degeneration. In dystrophic rats, MCs containing rhodopsin or rod outer segment (ROS) debris were found outside the outer retina at sites suggesting choroidal and ciliary egress. In conclusion, our data indicate that MC accumulation at injury sites is independent of CX3CR1 and precedes photoreceptor degeneration. The ectopic presence of rhodopsin-positive MCs suggests that CX3CR1 participates in MC egress from the outer retina. PMID- 18508132 TI - CXCR4 signaling in the regulation of stem cell migration and development. AB - The regulated migration of stem cells is a feature of the development of all tissues and also of a number of pathologies. In the former situation the migration of stem cells over large distances is required for the correct formation of the embryo. In addition, stem cells are deposited in niche like regions in adult tissues where they can be called upon for tissue regeneration and repair. The migration of cancer stem cells is a feature of the metastatic nature of this disease. In this article we discuss observations that have demonstrated the important role of chemokine signaling in the regulation of stem cell migration in both normal and pathological situations. It has been demonstrated that the chemokine receptor CXCR4 is expressed in numerous types of embryonic and adult stem cells and the chemokine SDF-1/CXCL12 has chemoattractant effects on these cells. Animals in which SDF-1/CXCR4 signaling has been interrupted exhibit numerous phenotypes that can be explained as resulting from inhibition of SDF-1 mediated chemoattraction of stem cells. Hence, CXCR4 signaling is a key element in understanding the functions of stem cells in normal development and in diverse pathological situations. PMID- 18508133 TI - CX3CL1 and CX3CR1 in the GL261 murine model of glioma: CX3CR1 deficiency does not impact tumor growth or infiltration of microglia and lymphocytes. AB - Human glioblastoma multiforme (GBM) is the most malignant form of human brain tumors. A characteristic of GBM is the marked presence of tumor infiltrated microglia/macrophages and lymphocytes. The goal of this study was directed toward understanding the role of the chemokine system CX3CL1 and its receptor CX3CR1 in the GL261 murine model of malignant glioma. In situ hybridization analysis identified CX3CL1 and CX3CR1 expression in GL261 tumors. The impact of CX3CR1 deletion on the growth of intracranial GL261 gliomas and associated immune cell infiltration was evaluated in CX3CR1 gene-disrupted C57BL/6 mice. A slight increase in the tumor growth rate in CX3CR1-/- mice was evident with similar numbers of microglia and CD4+, CD8+, FoxP3+, or Ly49G2+ lymphocytes within tumors established in CX3CR1 +/- and -/- mice. These data indicate that CX3CR1 has little or no effects on either gliomagenesis or the migration of microglia and lymphocytes into GL261 tumors. PMID- 18508134 TI - Establishment of reverse genetics system of betanodavirus for the efficient recovery of infectious particles. AB - Betanodaviruses, a member of the family Nodaviridae, have small positive-stranded bipartite RNA genomes and are the causal agent of viral nervous necrosis in marine-farmed fish. To facilitate the study of betanodavirus, infectious cDNA clones of its two genomic RNAs were generated. The full-length cDNA of the new Redspotted grouper nervous necrosis virus strain (SG2001Nag) RNA1 and RNA2 were co-transcribed by T7 RNA polymerase in baby hamster kidney cells expressing T7 RNA polymerase. The transcription of precise viral RNAs from cDNAs neither lead to viral protein synthesis nor the production of infectious particles. However, the additional two guanine residues following T7 promoter increased the transcription of viral RNAs from cDNAs, and 1.0 x 10(6)TCID(50)/ml of infectious particles was collected from the transfected cells. The ability to reproduce the entire life cycle of betanodavirus from cDNA clones by this reverse genetics system would therefore facilitate a further analysis of the mechanism of betanodavirus RNA replication, structure, and assembly. These findings may thus help in the future development of a betanodavirus vaccine. PMID- 18508135 TI - A practicable laboratory flow-through exposure system for assessing the health effects of effluents in fish. AB - The knowledge that exposure to estrogenic wastewater treatment work (WwTW) effluents induces a range of reproductive abnormalities in fish has highlighted the need to understand the wider health effects of effluents. Access to laboratory-based testing systems for WwTW effluents could greatly facilitate this endeavour. In this investigation, a laboratory-based test system was developed and applied for WwTW effluents using fathead minnows (Pimephales promelas). Sexually maturing fathead minnows were exposed, under flow-through conditions in the laboratory, for up to 21 days to graded concentrations of effluent from three different UK (temperate) WwTWs. The stability of the estrogenic component within the test system was assessed via measurements for estradiol and estrone concentrations in the effluent, and through determining estrogenic responses in an in vitro recombinant yeast estrogen screen (rYES) and in fish (plasma vitellogenin induction). The estrogen component of the effluents was stable within the holding system used (chilled <10 degrees C) for up to 7 days and measured concentrations of estradiol and estrone were shown to differ by less than 20% between the first and final day of use for each batch of effluent. Total estrogenic activity as measured in the rYES was found to be more variable (up to 66% variance between measurements for the two time points) but there was no consistent trend for a reduction in estrogenic activity. Vitellogenin was induced in males in a concentration-dependent manner and the magnitude of the response observed was proportional to the average measured concentrations of estradiol and estrone in the exposure effluent. The system described, thus, provides a robust test method for evaluating the estrogenic effects of temperate WwTW effluents that could be further applied to assess wider health effects, including population-relevant endpoints such as reproduction, using model OECD warm-water fish species such as the fathead minnow. PMID- 18508136 TI - Molecular characterization of cytochrome P450 1A and 3A and the effects of perfluorooctanoic acid on their mRNA levels in rare minnow (Gobiocypris rarus) gills. AB - Perfluorooctanoic acid (PFOA), a potentially toxic perfluorinated compound (PFC), has been widely disseminated in the environment. In the present study, rare minnows (Gobiocypris rarus) exposed to PFOA exhibited histopathological gill damage, including epithelial hyperplasia of the lamellae, inflammatory cell infiltration, and lamellar fusion. Cytochrome P450s (CYPs) play a central role in the metabolism and biotransformation of a wide range of endogenous substrates and foreign compounds. Thus, we studied the CYPs and the effects of waterborne PFOA on their corresponding mRNA levels in the gills of rare minnows. Two novel CYP cDNAs (CYP1A and CYP3A) were identified in rare minnow and their mRNAs were ubiquitously expressed in all tissues examined. Upregulation of CYP3A mRNA was observed in the gills of male rare minnows exposed to 30mg/L PFOA, while no significant changes occurred in exposed females. In contrast, down regulation of CYP1A mRNA was detected in the gills of male and female minnows exposed to PFOA. However, the effect of PFOA on gill mRNA levels of their potential regulators, aryl hydrocarbon receptor (AhR) for CYP1A, and pregnane X receptor (PXR) for CYP3A, were not consistent with the observed effects of PFOA on the corresponding CYP mRNA concentrations. This suggests a different or more complex transcriptional regulation of CYP expression following PFOA exposure. PMID- 18508137 TI - The RhopH complex is transferred to the host cell cytoplasm following red blood cell invasion by Plasmodium falciparum. AB - The high-molecular mass rhoptry protein complex (PfRhopH), which comprises three distinct gene products, RhopH1, RhopH2, and RhopH3, is known to be secreted and transferred to the parasitophorous vacuole membrane upon invasion of a red blood cell by the malaria parasite Plasmodium falciparum. Here we show that the merozoite-acquired RhopH complex is also transferred to defined domains of the red blood cell cytoplasm, and possibly transiently associated with Maurer's clefts. This is the first report of trafficking in the host cell cytoplasm for P. falciparum rhoptry proteins secreted upon red blood cell invasion. Based on its newly identified sub-cellular location and the phenotype of RhopH1 mutants, we propose that the RhopH complex participate in the assembly of the cytoadherence complex. PMID- 18508138 TI - Bed-exit alarm effectiveness. AB - This study describes the accuracy of two types of bed-exit alarms to detect bed exiting body movements: pressure-sensitive and a pressure-sensitive combined with infrared (IR) beam detectors (dual sensor system). We also evaluated the occurrence of nuisance alarms, or alarms that are activated when a participant does not attempt to get out of bed. Fourteen nursing home residents were directly observed for a total of 256 nights or 1636.5h; an average of 18.3+/-22.3 (+/ S.D.) nights/participant for an average of 6.4+/-1.2 h/night. After adjusting for body movements via repeated measures, Poisson regression modeling, the least squares adjusted means (LSM) show a marginally significant difference between the type of alarm groups on the number of true positives (NTP) (mean/S.E.M.=0.086/1.617) for pressure-sensitive versus dual sensor alarm (0.593/1.238; p=0.0599) indicating that the dual sensor alarm may have a higher NTP. While the dual sensor bed-exit alarm was more accurate than the pressure sensitive alarm in identifying bed-exiting body movements and reducing the incidence of false alarms, false alarms were not eliminated altogether. Alarms are not a substitute for staff; adequate staff availability is still necessary when residents need or wish to exit bed. PMID- 18508139 TI - Asynchronous normal regional left ventricular function assessed by speckle tracking echocardiography: appearances can be deceptive. AB - BACKGROUND: Speckle tracking echocardiography (STE) is an angle independent method with high temporal resolution, which offers quantification of regional left ventricular (LV) wall motion. We studied radial and longitudinal LV wall motion by STE in healthy subjects with normal wall motion analysis (WMA) by eye balling. MATERIALS AND METHODS: Eighteen healthy subjects were studied. We acquired parasternal short and apical long axis projections to determine the basal, mid and apical radial and longitudinal functions. At each level we measured; (I) radial and longitudinal peak displacement and displacement at aortic valve closure (AVC) and (II) the time interval from the Q-wave to the AVC and peak displacement. RESULTS: WMA indicated normal wall motion in all subjects. The mean peak radial displacement varied in different segments (range 3.9-9.8 mm) with highest values in the mid-level (6.9+/-1.5 mm), compared to basal level (5.9+/-1.0 mm, p<0.01) and apical level (5.4+/-1.0 mm, p<0.001). The time from Q wave to AVC was 393 ms and in 89% of the analysed segments peak radial displacement occurred after AVC, thus mean peak radial displacement occurred 60 ms after AVC. The peak longitudinal amplitude was more synchronous with respect to AVC and with the highest amplitudes found in the two basal segments. CONCLUSIONS: In normal LV function, significant differences in peak displacement exist between segments at various LV levels using STE. In addition, in early diastole, significant discrepancy occurs between radial and longitudinal time of peak displacement, suggesting a shape change. Finally, while radial displacement was highest at mid-cavity level longitudinal displacement was highest at basal level. PMID- 18508140 TI - Incidence and predictors of drug-eluting stent fractures in long coronary disease. AB - BACKGROUND: Stent fractures after drug-eluting stent (DES) implantation have not been evaluated sufficiently in patients with long coronary artery disease. METHODS: This study comprised of 415 patients, who were enrolled in the Long-DES II study and had a complete serial angiography both before and after procedure and also at follow-up. The lesions were > or =25 mm in length and were randomly treated with sirolimus-eluting stents (SES, 210 lesions) or paclitaxel-eluting stent (205 lesions). RESULTS: DES fracture was identified in 7 lesions (1.7%): 1 minor, 3 moderate, and 3 severe fractures. Most of the fractures occurred in patients who received SES (85.7%) and in the right coronary artery (RCA) lesions (71.4%). Lesions with fracture had a smaller minimal lumen diameter before procedure than lesions without fracture (0.38+/-0.55 vs. 0.71+/-0.46 mm, p=0.043). However, acute gain (2.28+/-0.39 vs. 1.44+/-0.60 mm, p=0.001) and late loss (0.81+/-0.49 vs. 0.42+/-0.50 mm, p=0.033) in analysis segment were greater in lesions with fracture. By multivariate analysis, the independent predictor of fracture was the RCA lesion (Odds ratio, 7.81; 95% CI, 1.45 approximately 42.04; p=0.017). Although one patient had an intermediate angiographic narrowing at the fracture site, there was no adverse cardiac event related with fracture. CONCLUSIONS: The incidence of stent fracture in long DES implantation was not common and was associated with SES implantation or RCA lesions. Fortunately, the clinical prognosis of DES fracture was somewhat benign. PMID- 18508141 TI - Percutaneous revascularization in patients with previous coronary artery bypass graft surgery. Immediate and 1-year clinical outcomes. AB - BACKGROUND: An increasing number of patients undergoing percutaneous interventions (PI) have experienced previous coronary artery bypass graft surgery (CABG). However, the impact of PI on outcomes in such patients is currently unclear. We evaluated the immediate and 1-year clinical outcomes of post-CABG patients who underwent PI in a tertiary center. METHODS: From January-2005 to September-2006, 91 consecutive post-CABG patients underwent 197 stent implantations (84% drug-eluting stents) for 154 lesions. 58% were treated in the native coronary arteries, 34% in the grafts and 8% in both type of vessels. Major adverse cardiac events (MACE) were recorded in-hospital and at 1-year follow-up. RESULTS: Procedural success rate was 95.6%. In-hospital MACE rate was 3.3%. At 1 year, the incidence of MACE was 18.6%: death occurred in 5.4% of the patients, myocardial infarction in 2.2%, and 10.9% of the patients underwent repeat revascularization (target lesion revascularization was required in 5.4%). Multivariate analysis revealed left ventricular ejection fraction <50% (OR 4.6, 95% CI 1.8 to 7.5, p=0.01) and multivessel intervention (OR 2.7, 95% CI 1.2 to 4.5, p=0.03) to be independent predictors of MACE at 1 year. CONCLUSIONS: Immediate results showed the safety and efficacy of percutaneous revascularization in post-CABG patients. The relatively low risk need for target lesion revascularization obtained is encouraging. Independent predictors of MACE at 1 year were left ventricular disfunction and multivessel intervention. PMID- 18508142 TI - Cardiac implications of Lyme disease, diagnosis and therapeutic approach. AB - Lyme is a tick-borne disease. The genetic diversity of Borreliae its distribution worldwide and its epidemiology have been related to different clinical manifestations. Carditis is a rare manifestation of Lyme disease. The commonest abnormality is atrioventricular block of various degrees, though other rhythm abnormalities have been reported. Pericarditis, myocarditis, cardiomyopathy and degenerative valvular disease have been associated with B. burgdorferi. Temporary pacing might be required in unstable patients. The majority of the conduction disturbances have a benign prognosis, if the infectious agent is identified and treated appropriately. PMID- 18508143 TI - Left ventricular apical ballooning syndrome: prevalence, clinical characteristics and pathogenetic mechanisms in a European population. AB - BACKGROUND: Left ventricular apical ballooning syndrome (LVABS) is a cardiac syndrome mimicking acute myocardial infarction, whose prevalence in western populations and pathogenesis are not yet well defined. The aim of the study was to assess its prevalence, clinical characteristics and pathophysiological mechanisms in a European population of myocardial infarction patients. METHODS: Of a series of 1457 patients with acute myocardial infarction 18 fulfilled the diagnostic criteria for LVABS. To evaluate the pathogenetic mechanisms we studied coronary blood flow with TIMI flow grade and corrected TIMI frame count (CTFC) in all patients and performed provocative testing with ergonovine and dobutamine echocardiography in 14. RESULTS: All patients were women aged 72+/-9 years. A triggering event was identifiable in 39% of cases. LV ejection fraction in the acute phase was 46+/-5%. No deaths or major complications occurred during hospitalization. Response to ergonovine was negative in all 14 patients and dobutamine induced a dynamic LV obstruction in 4/14 (28%). Mean CTFC was abnormally prolonged in all 3 major coronary arteries and 16/18 patients (89%) had an abnormal CTFC in >/=1 coronary vessel. No cardiac deaths occurred during follow-up and 1 patient only had a recurrence. CONCLUSIONS: The prevalence of LVABS is 1.2% among all patients with acute myocardial infarction, but rises to 4.9% in women. Short- and long-term prognosis is good. Abnormal CTFC suggests the presence of a coronary microvascular dysfunction, while dynamic LV obstruction can contribute to the development of LVABS in a minority of patients. PMID- 18508144 TI - A review of geospatial and ecological factors affecting disease spread in wild pigs: considerations for models of foot-and-mouth disease spread. AB - Around the world, wild boar or feral pigs are infected by a range of infectious organisms with important, productivity, public health or economic consequences. Consequently, the potential role of wild pigs in outbreaks of important exotic diseases, like foot-and-mouth disease (FMD), has been a significant consideration in many countries. Disease modelling is one means to study the epidemiology of disease and has been used to assess the potential role of wild pigs in FMD incursions. Many of these models have been strategic in nature. They have contributed to a broad understanding of disease control in wild pigs (e.g. the concept of threshold densities and the need to cull pigs below this density for disease fadeout to occur), but have not incorporated many of the key drivers affecting disease behaviour. Some of these drivers include important ecological, behavioural and geospatial relationships, such as interaction between different host species and the distribution, density and connectivity of pig populations. New approaches to modelling disease spread such as spatial simulation models use spatial data and explicitly incorporate geospatial relationships. These approaches can provide useful quantitative models that can be used to explore mitigation strategies under specific disease outbreak conditions. However, to date, most studies have been limited by inadequate data, and computational issues or have not explored mitigation strategies. To inform management strategies for emergency epidemics such as FMD in wild pigs, there is scope to further develop and use models to explore a range of incursion scenarios and investigate the efficacy of different mitigation strategies. PMID- 18508145 TI - Acute dextro-amphetamine administration does not alter brain myo-inositol levels in humans and animals: MRS investigations at 3 and 18.8 T. AB - The pathophysiological underpinnings of bipolar disorder are not fully understood. However, they may be due in part to changes in the phosphatidylinositol second messenger system (PI-cycle) generally, or changes in myo-inositol concentrations more specifically. Dextro-amphetamine has been used as a model for mania in several human studies as it causes similar subjective and physiological symptoms. We wanted to determine if dextro-amphetamine altered myo inositol concentrations in vivo as it would clearly define a mechanism linking putative changes in the PI-cycle to the subjective psychological changes seen with dextro-amphetamine administration. Fifteen healthy human volunteers received a baseline scan, followed by second scan 75 min after receiving a 25 mg oral dose of dextro-amphetamine. Stimulated echo proton magnetic resonance spectroscopy (MRS) scans were preformed at 3.0 Tesla (T) in the dorsal medial prefrontal cortex (DMPFC). Metabolite data were adjusted for tissue composition and analyzed using LCModel. Twelve adult male rats were treated acutely with a 5-mg/kg intraperitoneal dose of dextro-amphetamine. After 1 h rats were decapitated and the brains were rapidly removed and frozen until dissection. Rat brains were dissected into frontal, temporal, and occipital cortical areas, as well as hippocampus. Tissue was analyzed using a Varian 18.8 T spectrometer. Metabolites were identified and quantified using Chenomx Profiler software. The main finding in the present study was that myo-inositol concentrations in the DMPFC of human volunteers and in the four rat brain regions were not altered by acute dextro amphetamine. While it remains possible that the PI-cycle may be involved in the pathophysiology of bipolar disorder, it is not likely that the subjective and physiological of dextro-amphetamine are mediated, directly or indirectly, via alternations in myo-inositol concentrations. PMID- 18508146 TI - Occurrence and genotypes of Campylobacter in broiler flocks, other farm animals, and the environment during several rearing periods on selected poultry farms. AB - On 15 Swiss poultry farms, broiler flocks, other farm animals, and the environment were examined during consecutive rearing periods to investigate the occurrence and genetic diversity of Campylobacter. Of the 5154 collected samples, 311 (6%) from 14 farms were Campylobacter positive by culture. Amongst the positive samples, 228 tested positive for Campylobacter jejuni and 92 for Campylobacter coli. Positive samples originated from broilers, the broiler houses, cattle, pigs, bantams, laying hens, a horse, and a mouse. Feed, litter, flies, and the supply air to the broiler house tested negative. By flagellin gene typing (fla-RFLP) and pulsed-field gel electrophoresis (PFGE), 917 Campylobacter isolates were genotyped. Additionally, amplified fragment length polymorphism (AFLP) analysis was performed on 15 assorted strains. On eight farms, matching genotypes were isolated from broiler flocks and other farm animals: Certain genotypes from cattle (farms H, K, L, and M), pigs (farms D and P), or laying hens (farm L) were subsequently found in the broiler flocks, whereas other genotypes initially present in the broiler flocks turned up in cattle (farms A, D, and O). These results emphasize the importance of other farm animals on poultry farms for broiler flock colonization. Indications of persistent contamination of the broiler house were evident on four farms (C, D, I, and L) where matching genotypes were detected in consecutive broiler flocks, but not concurrently in other samples. By fla-RFLP, PFGE, and confirmed by AFLP, some genotypes proofed to be identical across different farms. PMID- 18508147 TI - Coating didodecyldimethylammonium bromide onto Au nanoparticles increases the stability of its complex with DNA. AB - The stability of the complex of cationic lipid with nucleic acid, especially when facing serum, is crucial for the efficiency of gene delivery. Here, we demonstrated that the stability of the complex of didodecyldimethylammonium bromide (DDAB, a cationic lipid) with DNA in the presence of serum dramatically increased after coating DDAB onto the surface of the gold nanoparticles. The stability of the complex was demonstrated with dye intercalation assay, and agarose gel electrophoresis. The process of the interaction was characterized with UV-vis spectra and the morphology of the complex was observed with atomic force microscope (AFM). Cell viability assays demonstrated that the cytotoxicity of DDAB was also decreased. PMID- 18508149 TI - Experimental diabetes enhances Ca2+ mobilization and glutamate exocytosis in cerebral synaptosomes from mice. AB - The present study was conducted to investigate the effects of the diabetic condition on the Ca(2+) mobilization and glutamate release in cerebral nerve terminals (synaptosomes). Diabetes was induced in male mice by intraperitoneal injection of streptozotocin. Cytosolic free Ca(2+) concentration ([Ca(2+)](i)) and glutamate release in synaptosomes were determined using fura-2 and enzyme linked fluorometric assay, respectively. Diabetes significantly enhanced the ability of the depolarizing agents K(+) and 4-aminopyridine (4-AP) to increase [Ca(2+)](i). In addition, diabetes significantly enhanced K(+)- and 4-AP-evoked Ca(2+)-dependent glutamate release. The pretreatment of synaptosomes with a combination of omega-agatoxin IVA (a P-type Ca(2+) channel blocker) and omega conotoxin GVIA (an N-type Ca(2+) channel blocker) inhibited K(+)- or 4-AP-induced increases in [Ca(2+)](i) and Ca(2+)-dependent glutamate release in synaptosomes from the control and diabetic mice to a similar extent, respectively. These results indicate that diabetes enhances a K(+)- or 4-AP-evoked Ca(2+)-dependent glutamate release by increasing [Ca(2+)](i) via stimulation of Ca(2+) entry through both P- and N-type Ca(2+) channels. PMID- 18508150 TI - Coated-platelet levels in patients with Type 1 and with Type 2 diabetes mellitus. AB - Coated-platelet levels were quantified in 58 people with Type 1 diabetes, 90 with Type 2 diabetes, and 54 non-diabetic controls. In diabetes high coated-platelet levels were related to smoking and glucose control drugs, but not to glycaemia or other drugs. Prospective studies should evaluate coated-platelets and complications and drug effects. PMID- 18508151 TI - Drug formulary decision-making in two regional health authorities in British Columbia, Canada. AB - OBJECTIVES: Growing pharmaceutical demands challenge healthcare organizations to set drug funding priorities (i.e. establish a formulary list). This responsibility typically rests with pharmacy and therapeutics (P&T) committees, yet how the process transpires within regional health authorities is unclear. The purpose of this study was to construct an explanatory model of drug formulary priority-setting as it occurs within regional health authorities. METHODS: A grounded theory approach was employed to study the practices of two regional health authority P&T committees in British Columbia, Canada. Data sources spanned committee documents, meeting observations (n=4), and semi-structured interviews with committee members (n=15). Data analysis involved coding using the constant comparative technique and writing analytic memos. RESULTS: Regional P&T committees engaged in two activities related to drug formulary priority-setting: developing auto-substitution policies and reviewing drug addition requests. Four processes were central to decision-making: (i) negotiating margins of therapeutic advantage; (ii) seeking value for the resources allocated; (iii) interfacing between community and institutional settings; (iv) situating decisions within an organizational context. CONCLUSIONS: Findings highlight opportunities for institutions to improve the fairness of agenda-setting practices, and for additional collaboration between policy-makers who prioritize drugs for publicly funded formularies applicable to institutional versus community settings. PMID- 18508152 TI - Artificial neural network prediction of clozapine response with combined pharmacogenetic and clinical data. AB - Although one third to one half of refractory schizophrenic patients responds to clozapine, however, there are few evidences currently that could predict clozapine response before the use of the medication. The present study aimed to train and validate artificial neural networks (ANN), using clinical and pharmacogenetic data, to predict clozapine response in schizophrenic patients. Five pharmacogenetic variables and five clinical variables were collated from 93 schizophrenic patients taking clozapine, including 26 responders. ANN analysis was carried out by training the network with data from 75% of cases and subsequently testing with data from 25% of unseen cases to determine the optimal ANN architecture. Then the leave-one-out method was used to examine the generalization of the models. The optimal ANN architecture was found to be a standard feed-forward, fully-connected, back-propagation multilayer perceptron. The overall accuracy rate of ANN was 83.3%, which is higher than that of logistic regression (LR) (70.8%). By using the area under the receiver operating characteristics curve as a measure of performance, the ANN outperformed the LR (0.821+/-0.054 versus 0.579+/-0.068; p<0.001). The ANN with only genetic variables outperformed the ANN with only clinical variables (0.805+/-0.056 versus 0.647+/-0.066; p=0.046). The gene polymorphisms should play an important role in the prediction. Further validation of ANN analysis is likely to provide decision support for predicting individual response. PMID- 18508153 TI - Development of novel statistical reconstruction algorithms for poly-energetic X ray computed tomography. AB - A beam-hardening effect is a common problem affecting the quantitative aspects of X-ray computed tomography (CT). We have developed two statistical reconstruction algorithms for poly-energetic X-ray CT that can effectively reduce the beam hardening effect. Phantom tests were used to evaluate our approach in comparison with traditional correction methods. Unlike previous methods, our algorithm utilizes multiple energy-corresponding blank scans to estimate the attenuation map for a particular energy spectrum. Therefore, our algorithm is an energy selective reconstruction. In addition to benefits over other statistical algorithms for poly-energetic reconstruction, our algorithm has the advantage of not requiring prior knowledge of the object material, the energy spectrum of the source and the energy sensitivity of the detector. The results showed an improvement in coefficient of variation, uniformity and signal-to-noise ratio; overall, this novel approach produces a better beam-hardening correction. PMID- 18508154 TI - Porous silicon in drug delivery devices and materials. AB - Porous Si exhibits a number of properties that make it an attractive material for controlled drug delivery applications: The electrochemical synthesis allows construction of tailored pore sizes and volumes that are controllable from the scale of microns to nanometers; a number of convenient chemistries exist for the modification of porous Si surfaces that can be used to control the amount, identity, and in vivo release rate of drug payloads and the resorption rate of the porous host matrix; the material can be used as a template for organic and biopolymers, to prepare composites with a designed nanostructure; and finally, the optical properties of photonic structures prepared from this material provide a self-reporting feature that can be monitored in vivo. This paper reviews the preparation, chemistry, and properties of electrochemically prepared porous Si or SiO2 hosts relevant to drug delivery applications. PMID- 18508156 TI - Renal uptake and metabolism of radiopharmaceuticals derived from peptides and proteins. AB - Radiolabeled anti-CD20 antibodies have demonstrated impressive efficacy in the treatment of relapsed non-Hodgkin lymphoma. This encourages the treatment of solid tumor with radiolabeled antibody fragments and peptides. However, both preclinical and clinical studies revealed that persistent localization of radioactivity in the kidney constitutes a major obstacle that compromises therapeutic efficacy. Recent extensive studies show that long residence times of radiolabeled end products from lysosomes are responsible for the renal radioactivity levels. Recent studies have also elucidated the involvement of megalin-cubilin in renal tubular reabsorption of radiolabeled antibody fragments and peptides. In light of these findings, efforts are being made to block tubular reabsorption of radiolabeled antibody fragments and peptides by competitive inhibitors, charge modification, and PEGylation. An interposition of an enzyme cleavable linkage between antibody fragments and radiolabels would constitute an alternative approach to reduce renal radioactivity levels. Recent findings of these studies will be described. PMID- 18508155 TI - Use of antibodies and immunoconjugates for the therapy of more accessible cancers. AB - There are currently 6 unconjugated antibodies and 3 immunoconjugates approved for use in the United States in a variety of cancers, with a considerable number of new agents in clinical testing and preclinical development. Unconjugated antibodies alone can be effective, but more often, antibodies need to be combined with chemotherapy, which enhances the efficacy of the standard treatment. Immunoconjugates tend to be more effective than their unconjugated counterparts, but their increased toxicity often restricts when and how they are used. In order to improve efficacy, a number of immunoconjugates are being examined in settings where the disease is more easily accessible, such as leukemias, or within compartments that allow easier and more direct access to the tumor, such as in the peritoneal cavity or brain, or both locally and systemically, in adjuvant situations, where the disease burden has been reduced by some other means, and with the main goal of these treatments being to kill residual disease. PMID- 18508157 TI - Multifunctional magnetic nanoparticles for targeted imaging and therapy. AB - Magnetic nanoparticles have become important tools for the imaging of prevalent diseases, such as cancer, atherosclerosis, diabetes, and others. While first generation nanoparticles were fairly nonspecific, newer generations have been targeted to specific cell types and molecular targets via affinity ligands. Commonly, these ligands emerge from phage or small molecule screens, or are based on antibodies or aptamers. Secondary reporters and combined therapeutic molecules have further opened potential clinical applications of these materials. This review summarizes some of the recent biomedical applications of these newer magnetic nanomaterials. PMID- 18508158 TI - Docetaxel plus gemcitabine as front-line chemotherapy in elderly patients with lung adenocarcinomas: a multicenter phase II study. AB - BACKGROUND: The docetaxel/gemcitabine (DG) combination is an active and well tolerated regimen against non-small cell lung cancer (NSCLC). A phase II study was conducted in order to evaluate its efficacy in elderly patients with lung adenocarcinomas. METHODS: Chemotherapy-naive patients, aged > or =70 years, with locally advanced or metastatic lung adenocarcinomas and performance status (PS) < or =2 (ECOG) received gemcitabine 1100 mg/m(2) (days 1+8) and docetaxel 100 mg/m(2) (day 8) with rhG-CSF support. RESULTS: Seventy-seven patients were enrolled. One (1.3%) complete and 23 (29.9%) partial responses were achieved (intention to treat analysis: ORR 31.2%; 95% CI 20.82-41.51%) whereas tumor growth control was achieved in 53.3% of patients. The median TTP was 4.1 months, the median overall survival 9.4 months and the 1- and 2-year survival rate 37.9% and 10.7%, respectively. Grade 3-4 neutropenia occurred in 18.2% and febrile neutropenia in 3 (3.9%) patients. Non-haematological toxicity was mild with grade 2-3 asthenia occurring in 22.1% patients. CONCLUSIONS: The DG regimen is an active and well-tolerated front-line chemotherapy for elderly patients with lung adenocarcinomas and merits further evaluation in prospective randomized trials. PMID- 18508159 TI - Influence of Na2S on the degradation kinetics of CCl4 in the presence of very pure iron. AB - This paper presents the results of kinetic studies to investigate the effect of FeS film formation on the degradation rate of CCl(4) by 99.99% pure metallic iron. The film was formed by submersing metallic iron grains in an oxygen free HCO(3)(-)/CO(3)(2-) electrolyte solution. When the grains had reached a quasi steady-state value of the corrosion potential, Na(2)S((aq)) was injected. Upon injection, a microm thick poorly crystalline FeS film formed immediately on the iron surface. Over time, the iron became strongly corroded and both the FeS film and the metallic iron grains began to crack leading to exposure of bare metallic iron to the solution. The effect of the surface film on the degradation rate of CCl(4) was investigated following four periods of aging, 1, 10, 30, and 60 days. Relative to the controls, the 1-day sulfide-aged iron showed a substantial decrease in rate of degradation of CCl(4.) However, over time, the rate of degradation increased and surpassed the degradation rate obtained in the controls. It has been proposed that CCl(4) is reduced to HCCl(3) by metallic iron by electron transfer. The FeS film is substantially less conducting than the bulk iron metal or non-stoichiometric magnetite and from the results of this study, greatly decreases the rate of CCl(4) degradation relative to iron that has not been exposed to Na(2)S. However, continued aging of the FeS film results in breakdown and stress-induced cracking of the film, followed by dissolution and cracking of the iron itself. The cracking of the bulk iron is believed to be a consequence of hydrogen embrittlement, which is promoted by sulfide. The increase in CCl(4) degradation rate, as the FeS films age, suggests that the process of hydrogen cracking increases the surface area available for charge transfer. PMID- 18508160 TI - Metabolic effects of chronic obestatin infusion in rats. AB - Obestatin is purported to be a peptide hormone encoded in preproghrelin. We studied the metabolic effects of continuous infusion of obestatin via subcutaneously implanted osmotic mini-pumps. Administration of up to 500nmol/kg body weight/day obestatin did not change 24h cumulative food intake or body weight in rats. Similarly, no effects were observed when obestatin was infused at 1000nmol/kg body weight/day for seven days. This dose of obestatin infused during a 24h fast did not alter weight loss, suggesting that obestatin has no effect on energy expenditure, and this dose did not alter glucose or insulin responses during an IPGTT. Obestatin was originally proposed to interact with GPR39 and subsequently the receptor for GLP-1. While both receptors are expressed in pancreatic islets, incubation with obestatin did not alter insulin release from islets in vitro. Moreover, obestatin did not bind to INS-1 beta-cells or HEK cells overexpressing GLP-1 receptors or displace GLP-1 binding to these cells. Our findings do not support the concept that obestatin is a hormone with metabolic actions. PMID- 18508161 TI - Synthesis and antimicrobial activities of some new triazolothiadiazoles bearing 4 methylthiobenzyl moiety. AB - A series of substituted triazolothiadiazoles have been synthesized by condensing 4-amino-3-[4-methylthiobenzyl]-4H-1,2,4-triazole-5-thiol (5) with substituted aryl furoic acids/aromatic acids in the presence of POCl3. The triazole (5) was obtained by the fusion of 4-methylthiophenyl acetic acid with thiocarbohydrazide. The structures of newly synthesized compounds are characterized by elemental analysis, IR, 1H NMR and mass spectroscopic studies and were screened for their antimicrobial activities. The preliminary results revealed that some of the compounds exhibited promising antimicrobial activities. PMID- 18508162 TI - [Autoimmune myelofibrosis with dermatomyositis]. AB - We report a second observation of autoimmune myelofibrosis associated with an inflammatory myositis in a 30-year-old female. The links between myelofibrosis and autoimmunity are discussed. PMID- 18508163 TI - Neurovirulence evaluation of Venezuelan equine encephalitis (VEE) vaccine candidate V3526 in nonhuman primates. AB - Assessment of neurovirulence is a standard test for vaccines derived from virulent neurotropic viruses. This study evaluated the potential neurovirulence of V3526, a live attenuated vaccine derived from a full-length infectious clone of Venezuelan equine encephalitis virus (VEEV) Trinidad donkey strain (TrD), a comparator VEEV vaccine (TC-83), TrD, and process control material (PCM) in juvenile rhesus macaques. Following intrathalamic/intraspinal (i.t./i.s. ) or subcutaneous (s.c.) inoculations, animals were observed for periods of 18, 91 or 181 days for paresis, paralysis, neurological disorders and other signs of clinical illness. Blood was collected for measurement of viremia, VEEV neutralizing antibodies, hematologic parameters, and liver enzymes. Gross necropsies and histopathological examinations were conducted with emphasis on detecting lesions in the brain and spinal cord. Elevated temperatures (1-2 degrees C) were noted in several of the TrD and vaccine inoculated animals on Day 6 following inoculation and mean temperatures for the V3526 i.t./i.s. and TC-83 groups were higher than PCM group throughout the study Day 18. No significant differences were seen for weight or clinical chemistry results between vaccine and PCM inoculated groups. Clinically significant signs (Grades 3 or 4) were noted in three of 21 V3526 i.t./i.s. and three of 12 TC-83 inoculated animals, however, these signs resolved within 3 weeks for all V3526 i.t./i.s. and for two of three TC-83 inoculated animals. At Day 18 extensive lesions indicative of a viral infection were seen in brain sections of all four TrD inoculated animals and one of seven V3526 i.t./i.s. inoculated animals. Only scattered lesions, characterized by foci of gliosis and vessels with perivascular inflammation, were found in the sections from four TC-83 and six V3526 i.t./i.s. inoculated animals. The minimal histological changes observed at Day 18 resolved to baseline levels by Day 181 comparable to the PCM group. V3526 was immunogenic and essentially nonneurovirulent when administered via the clinically relevant subcutaneous route. PMID- 18508164 TI - Addressing uncertainties in the ERICA Integrated Approach. AB - Like any complex environmental problem, ecological risk assessment of the impacts of ionising radiation is confounded by uncertainty. At all stages, from problem formulation through to risk characterisation, the assessment is dependent on models, scenarios, assumptions and extrapolations. These include technical uncertainties related to the data used, conceptual uncertainties associated with models and scenarios, as well as social uncertainties such as economic impacts, the interpretation of legislation, and the acceptability of the assessment results to stakeholders. The ERICA Integrated Approach has been developed to allow an assessment of the risks of ionising radiation, and includes a number of methods that are intended to make the uncertainties and assumptions inherent in the assessment more transparent to users and stakeholders. Throughout its development, ERICA has recommended that assessors deal openly with the deeper dimensions of uncertainty and acknowledge that uncertainty is intrinsic to complex systems. Since the tool is based on a tiered approach, the approaches to dealing with uncertainty vary between the tiers, ranging from a simple, but highly conservative screening to a full probabilistic risk assessment including sensitivity analysis. This paper gives on overview of types of uncertainty that are manifest in ecological risk assessment and the ERICA Integrated Approach to dealing with some of these uncertainties. PMID- 18508165 TI - Influence of solar UV radiation on the nitrogen metabolism in needles of Scots pine (Pinus sylvestris L.). AB - Needles of 20-year-old Scots pine (Pinus sylvestris L.) saplings were studied in an ultraviolet (UV) exclusion field experiment (from 2000 to 2002) in northern Finland (67 degrees N). The chambers held filters that excluded both UV-B and UV A, excluded UV-B only, transmitted all UV (control), or lacked filters (ambient). UV-B/UV-A exclusion decreased nitrate reductase (NR) activity of 1-year-old needles of Scots pines compared to the controls. The proportion of free amino acids varied in the range 1.08-1.94% of total proteins, and was significantly higher in needles of saplings grown under UV-B/UV-A exclusion compared to the controls or UV-B exclusion. NR activity correlated with air temperature, indicating a "chamber effect". The study showed that both UV irradiance and increasing temperature are significant modulators of nitrogen (N) metabolism in Scots pine needles. PMID- 18508166 TI - Mercury methylation by a microbial community from sediments of the Adour Estuary (Bay of Biscay, France). AB - In order to study the influence of microorganisms on the mercury biogeochemistry, the metal content and the structure of microbial communities were determined in sediments from stations along the Adour Estuary. The comparison of the bacterial communities and their distribution in function of the environmental parameters by Canonical Correspondence Analysis (CCA) revealed the influence of metals on the bacterial communities structure. Sediments where the bacterial communities are mostly influenced by methylmercury were incubated in slurries with or without mercury, under oxic and anoxic conditions. Methylmercury production was detected in the anoxic biotic slurries with a net methylation yield of 0.3% after 24 h. CCA based on T-RFLP profiles revealed the impact of mercury addition on the bacterial communities structure. In addition, 17 bacterial strains, mainly sulphate-reducing bacteria involved in mercury methylation, were isolated and identified. PMID- 18508167 TI - Sorption of phenanthrene by dissolved organic matter and its complex with aluminum oxide nanoparticles. AB - Intent of this study was to explore the potential application of polymerin, the polymeric, dissolved organic matter fraction from olive oil wastewaters, in technologies aimed at remediating hydrophobic organic compounds (HOCs) point source pollution. Phenanthrene binding with polymerin was investigated. Moreover, the effect of addition of micro and nanoscale aluminum oxides (Al2O3) was studied, as well as sorption of polymerin on the oxides. Phenanthrene binding capacity by polymerin was notably higher than the sorption capacities for both types of Al2O3 particles. Polymerin sorption on nanoparticles was nearly 100 times higher than microparticles. In a three-phase system, using microparticles, higher phenanthrene sorption was found by adding into water polymerin, oxides and phenanthrene simultaneously. In contrast, using nanoparticles, a considerable enhancement of phenanthrene sorption was shown by adding phenanthrene to a pre formed and dried polymerin-oxide complex. These findings support the application of polymerin, especially associated with Al2O3 nanoparticles, in remediation of water contaminated with HOCs. This work highlights the significant role of nanoparticles. PMID- 18508168 TI - Reductions in the tobacco specific nitrosamine (TSNA) content of tobaccos taken from commercial Canadian cigarettes and corresponding reductions in TSNA deliveries in mainstream smoke from such cigarettes. AB - Tobacco specific nitrosamines (TSNAs) are suspected to cause smoking-related neoplastic diseases. The change from direct-fired to indirect-fired barns (aka kilns) for curing bright (aka Virginia, flue-cured) tobaccos was made to reduce the TSNA concentrations. The effectiveness of such processes in reducing the deliveries of TSNAs to the users of the products should be monitored. However, it is difficult to assess the effects of this reduction on the TSNA levels in mainstream smoke when cigarette blends contain burley tobaccos and other blend components that can increase smoke TSNA concentrations. Canadian cigarettes made prior to and in the few years just after the conversion to indirect-fired curing should not be subject to such interferences. Thus, the TSNA content of tobaccos and mainstream smoke from six brands of Canadian cigarettes produced in 2003, 2004, and 2005 were determined. Reductions in NNK [4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone], the most important TSNA in flue-cured tobaccos, levels in the tobacco blends ranged from 60% to 85%. The corresponding reductions in mainstream smoke TSNA levels ranged from 59% to 72% (ISO smoking conditions) and 58-76% (Health Canada Intensive smoking conditions). These results show that other factors (microorganisms, nitrite levels) may be negating the TSNA reductions achieved by indirect-fired curing. PMID- 18508169 TI - Differences in influence patterns between groups predicting the adoption of a solar disinfection technology for drinking water in Bolivia. AB - The lack of safe drinking water is one of the major problems faced by developing countries. The consequences of contaminated water are diseases such as diarrhea, one of the main causes of infant mortality. Because of its simplicity, solar water-disinfection technology provides a good way of treating water at the household level. Despite its obvious advantages and considerable promotional activities, this innovation has had rather a slow uptake. We conducted a field survey in which 644 households in Bolivia were interviewed in order to gain insights on motivations that resulted in adopting the technology. The aim was to examine possible differences in the predictors for adopting this technology during the diffusion process using the theory of innovation diffusion. Our findings indicate that early adoption was predicted by increased involvement in the topic of drinking water and that adoption in the middle of the diffusion process was predicted by increased involvement by opinion leaders and by recognition of a majority who supported the technology. Finally, late adoption was predicted by recognition that a majority had already adopted. Suggestions for future promotional strategies are outlined. PMID- 18508170 TI - Interaction of working conditions, job satisfaction, and sickness absences: evidence from a representative sample of employees. AB - We study the predictors of sickness absences among 2800 Finnish workers responding to the cross-sectional Quality of Work Life Survey in 1997. The data contain detailed information on the prevalence of adverse working conditions at the workplace from a representative sample of wage and salary earners. We show by using recursive multivariate models that the prevalence of harms at the workplace is associated with job dissatisfaction and dissatisfaction with sickness absences. The policy lesson is that the improvement of working conditions should be an integral part of any scheme aimed at decreasing sickness absence. PMID- 18508171 TI - Disparities and distrust: the implications of psychological processes for understanding racial disparities in health and health care. AB - This paper explores the role of racial bias toward Blacks in interracial relations, and in racial disparities in health care in the United States. Our analyses of these issues focuses primarily on studies of prejudice published in the past 10 years and on health disparity research published since the report of the US Institute of Medicine (IOM) Panel on Racial and Ethnic Disparities in Health Care in 2003. Recent social psychological research reveals that racial biases occur implicitly, without intention or awareness, as well as explicitly, and these implicit biases have implications for understanding how interracial interactions frequently produce mistrust. We further illustrate how this perspective can illuminate and integrate findings from research on disparities and biases in health care, addressing the orientations of both providers and patients. We conclude by considering future directions for research and intervention. PMID- 18508172 TI - Longitudinal studies to detect genexenvironment interactions in common disease- bang for your buck? A commentary on Chaufan's "how much can a large population study on genes, environments, their interactions and common diseases contribute to the health of the American people?" (65:8, 1730-1741(2007)). PMID- 18508173 TI - Perceptual grouping and distance estimates in typical and atypical development: comparing performance across perception, drawing and construction tasks. AB - Perceptual grouping is a pre-attentive process which serves to group local elements into global wholes, based on shared properties. One effect of perceptual grouping is to distort the ability to estimate the distance between two elements. In this study, biases in distance estimates, caused by four types of perceptual grouping, were measured across three tasks, a perception, a drawing and a construction task in both typical development (TD: Experiment 1) and in individuals with Williams syndrome (WS: Experiment 2). In Experiment 1, perceptual grouping distorted distance estimates across all three tasks. Interestingly, the effect of grouping by luminance was in the opposite direction to the effects of the remaining grouping types. We relate this to differences in the ability to inhibit perceptual grouping effects on distance estimates. Additive distorting influences were also observed in the drawing and the construction task, which are explained in terms of the points of reference employed in each task. Experiment 2 demonstrated that the above distortion effects are also observed in WS. Given the known deficit in the ability to use perceptual grouping in WS, this suggests a dissociation between the pre-attentive influence of and the attentive deployment of perceptual grouping in WS. The typical distortion in relation to drawing and construction points towards the presence of some typical location coding strategies in WS. The performance of the WS group differed from the TD participants on two counts. First, the pattern of overall distance estimates (averaged across interior and exterior distances) across the four perceptual grouping types, differed between groups. Second, the distorting influence of perceptual grouping was strongest for grouping by shape similarity in WS, which contrasts to a strength in grouping by proximity observed in the TD participants. PMID- 18508174 TI - Different effects of PCB101, PCB118, PCB138 and PCB153 alone or mixed in MCF-7 breast cancer cells. AB - Polychlorinated biphenyls (PCBs) are ubiquitous, persistent environmental contaminants that can be a potential health hazard. In the present study we analyzed the potential estrogenic effect in MCF-7 cells of four biologically relevant PCB congeners, alone or in mixtures, present in dairy products, vegetable oil and fish: PCB101, PCB118, PCB138 and PCB153. The mixture of four PCB was tested at seven different concentrations. We investigated the ability of these PCBs, alone or mixed, to induce cell proliferation, and the level of estrogen-regulated protein pS2, in human MCF-7 breast cancer cells. PCB153 (35 microM) stimulated cell proliferation from 48 h up to day 6, PCB118 (40 microM) only at 48 h, but PCB101 (45 microM) and PCB138 (15 microM) applied to the cells for 6 days had no effect. In contrast, the various concentrations of mixtures significantly reduced cell proliferation at different times. No change in pS2 levels was seen after treatment with the PCBs alone or mixed. In exploring the mechanism of these events, we found that PCB153 induced mitogen-activated protein kinase (MAPK) ERK1/2 at 4, 8 and 12 h, while the antiproliferative effect seemed to be related to an apoptotic action beginning at 12 h and ending at 48 h. These findings indicate that these PCBs alone or mixed have no estrogenic effect in MCF 7 cells, although PCB153 induce an ERK1/2-mediated mitogenic effect. On the contrary the mixture of PCBs induces an antiproliferative effect, ascribable to an apoptotic action. PMID- 18508175 TI - A 90-day toxicity study of L-asparagine, a food additive, in F344 rats. AB - L-asparagine is an amino acid listed as an existing food additive in Japan. The present 90-day toxicity study in F344/DuCrlCrj rats was conducted for safety assessment and to determine a no observed adverse effect level (NOAEL) of L asparagine. Groups of 10 males and 10 females were given the material at dose levels of 0%, 1.25%, 2.5% or 5% in diet for 90 days. During the experiment, there were no remarkable changes in general conditions and no deaths occurred in any group. Final body weights of male 5% and 1.25% groups were significantly decreased. There were also significant increases in relative organ weights of the brain, kidney and testis in 5% males. On serological examination, GLU, PL, K and ALT were increased significantly in 5% females, and GLU was increased significantly and CRN was decreased significantly in the female 1.25% group. However, histopathological examination did not reveal any significant variation in development of lesions among the groups. Changes in body and organ weights, as well as other parameters, were concluded to be due to treatment with 5% L asparagine. The NOAEL was determined to be 2.5% in the diet (males, 1.65 g/kg body weight/day; females, 1.73 g/kg body weight/day). PMID- 18508176 TI - The BIOSAFEPAPER project for in vitro toxicity assessments: preparation, detailed chemical characterisation and testing of extracts from paper and board samples. AB - Nineteen food contact papers and boards and one non-food contact board were extracted following test protocols developed within European Union funded project BIOSAFEPAPER. The extraction media were either hot or cold water, 95% ethanol or Tenax, according to the end use of the sample. The extractable dry matter content of the samples varied from 1200 to 11,800 mg/kg (0.8-35.5 mg/dm2). According to GC-MS the main substances extracted into water were pulp-derived natural products such as fatty acids, resin acids, natural wood sterols and alkanols. Substances extracted into ethanol particularly, were diisopropylnaphthalenes, alkanes and phthalic acid esters. The non-food contact board showed the greatest number and highest concentrations of GC-MS detectable compounds. The extracts were subjected to a battery of in vitro toxicity tests measuring both acute and sublethal cytotoxicity and genotoxic effects. None of the water or Tenax extracts was positive in cytotoxicity or genotoxicity assays. The ethanol extract of the non food contact board gave a positive response in the genotoxicity assays, and all four ethanol extracts gave positive response(s) in the cytotoxicity assays to some extent. These responses could not be pinpointed to any specific compound, although there appeared a correlation between the total amount of extractables and toxicity. PMID- 18508177 TI - Different effects of 26-week dietary intake of rapeseed oil and soybean oil on plasma lipid levels, glucose-6-phosphate dehydrogenase activity and cyclooxygenase-2 expression in spontaneously hypertensive rats. AB - We intended to determine whether or not dietary canola oil (CO) elevates plasma lipids and oxidative stress, since both of these are, possibly, related to the CO induced life shortening through exacerbation of hypertension-associated vascular lesions found in stroke-prone spontaneously hypertensive rats (SHRSP). Spontaneously hypertensive rats (SHR) were used in this study to avoid a potential bias in the results due to the irregular death by stroke seen in SHRSP. SHR were fed for 26 weeks on a chow containing either, 10 wt/wt% of CO or soybean oil (SO), i.e., the control. Elevated plasma lipids and glucose-6-phosphate dehydrogenase (G6PD) activation in the liver and erythrocyte were found in SHR fed CO compared to that fed SO, while anti-oxidative enzymes other than G6PD were not activated. The CO diet brought about significant vascular lesions in the kidney, in which abundant cyclooxygenase-2 (COX-2) positive foci were immunochemically located in the juxtaglomerular apparatus. These results suggest that dietary CO induces a hyperlipidemic condition, in which G6PD may serve as an NADPH provider, and aggravates genetic diseases in SHR (also, probably, in SHRSP). The increased COX-2 expression indicates a role of renin-angiotensin aldosterone system activation in the increased vascular lesions, whereas the effects of oxidative stress remain unclear. PMID- 18508178 TI - The effect of vitamin C on bisphenol A, nonylphenol and octylphenol induced brain damages of male rats. AB - Bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP) are endocrine-disrupting chemicals that has been shown to exert both toxic and estrogenic effects on mammalian cells. The aim of this study was to investigate if BPA, NP and OP induce oxidative stress on the brain tissue of male rats and if co-administration of vitamin C, an antioxidant, can prevent any possible oxidative stress. The male rats were divided into seven groups as control (vehicle), BPA, NP, OP, BPA+C, NP+C, OP+C. BPA, OP and NP (25 mg/(kg day)) were administrated orally to male Wistar rats for 45 days. In vitamin C co-administration groups (BPA+C, NP+C, OP+C), vitamin C (60 mg/(kg day)) were administrated orally along with BPA, OP and NP (25 mg/(kg day)) treatments. The rats in the control group received olive oil orally. The final body and absolute organ weights of treated rats did not show any significant difference when compared with the control group. Also, there were no significant difference in relative organ weights of BPA, NP, OP, BPA+C and NP+C groups when compared with control group. Only, relative organ weights were increased significantly in OP+C group compared with control group. Decreased levels of reduced glutathione (GSH) were found in the brains of BPA, NP, OP treated rats. The end product of lipid peroxidation, malondialdehyde (MDA), appeared at significantly higher concentrations in the BPA, NP, and OP treated groups when compared to the control group. On the other hand, there were no changes in the brain MDA and GSH levels of BPA+C, NP+C and OP+C groups compared with BPA, NP and OP treatment groups, respectively. In histopathologic examination, the vitamin C co-administrated groups had much more hyperchromatic cells in the brain cortex than that observed in the groups treated with only BPA, NP, and OP. The results of this study demonstrate that BPA, NP and OP generate reactive oxygen species that caused oxidative damage in the brain of male rats. In addition, vitamin C co-administration along with BPA, NP, and OP aggravates this oxidative damage in the brain of rats. PMID- 18508179 TI - Burning issues: early cooling of the brain after resuscitation using burn dressings. A proof of concept observation. AB - AIM OF THE STUDY: Early cooling of resuscitated patients improves neurological outcome. Out-of hospital initiation of cooling is uncommon however for mainly practical reasons. Using burn dressings in the out-of-hospital care could initiate brain cooling in an early stage and therefore be of value; the method is easily adaptable by ambulance crews. The influence of burn dressings on brain temperature is however unknown. We determined tympanic temperature changes as proxy for brain temperature in healthy volunteers after the application of cooling dressings to face and neck as a proof of concept study. METHOD: In 10 healthy human volunteers tympanic temperatures were measured in 30s intervals before, during and after the application of burn-dressings, special trauma burn care dressings that are designed for the acute treatment of skin burns (Burnshield emergency burn care sterile trauma burn dressings, Burnshield Ltd., Wadefield, South Africa) for the duration of 20min for each episode. RESULTS: In all study subjects the tympanic temperature was significantly lowered after 20min of the application of the burnshields. The mean difference between baseline and 2nd half of the exposure period was 0.43 degrees C (p<0.0001), ranging from 0.10 to 1.18 degrees C. CONCLUSION: Burn dressings could be of value in the early initiation of brain cooling in resuscitated patients. This study warrants further research to the effect of burnshield dressings on neurological activity and the effect on outcome after resuscitation. PMID- 18508180 TI - Should age be included as a component of track and trigger systems used to identify sick adult patients? AB - AIM OF STUDY: Few published "track and trigger systems" used to identify sick adult patients incorporate patient age as a variable. We investigated the relationship between vital signs, patient age and in-hospital mortality and investigated the impact of patient age on the function as predictors of in hospital mortality of the two most commonly used track and trigger systems. MATERIALS AND METHODS: Using a database of 9987 vital signs datasets, we studied the relationship between admission vital signs and in-hospital mortality for a range of selected vital signs, grouped by patient age. We also used the vital signs data set to study the impact of patient age on the relationship between patient triggers using the "MET criteria" and "MEWS", and in-hospital mortality. RESULTS: At hospital discharge, there were 9152 (91.6%) survivors and 835 (8.4%) non-survivors. As admission vital signs worsened, mortality increased for each age range. Where groups of patients had triggered a certain MET criterion, mortality was higher as patient age increased. Mortality varied significantly with age (p<0.05; Fishers exact test) for breathing rate >36breathsmin(-1), systolic BP<90mmHg and decreased conscious level. For each age group, mortality also increased as total MEWS score increased. As the number of simultaneously occurring MEWS abnormalities, or simultaneously occurring MET criteria, increased, mortality increased for each age range. CONCLUSIONS: Age has a significant impact on in-hospital mortality. Our data suggest that the inclusion of age as a component of these systems could be advantageous in improving their function. PMID- 18508181 TI - Use of the Laryngeal Mask Airway Supreme in pre-hospital difficult airway management. PMID- 18508182 TI - Cardiopulmonary resuscitation for neonates: evidence for compressions? PMID- 18508183 TI - "Win with your chin": an alternative to the "sniffing position" analogy for teaching optimal head-positioning with intubation. PMID- 18508184 TI - Cardiac arrest in the Emergency Department: a report from the National Registry of Cardiopulmonary Resuscitation. AB - BACKGROUND: Little is known about cardiac arrests (CA) in the Emergency Department (ED). The objective of this study was to determine the characteristics of ED CAs. METHODS: 60,852 adult, in-patient CA events in the National Registry of Cardiopulmonary Resuscitation were included. Multiple regression analysis compared ED CA with those occurring in the ICU, telemetry, or general floors. Subgroup analysis examined traumatic vs. non-traumatic ED CA and ED CA occurring after a successful pre-hospital resuscitation (recurrent) vs. primary ED event. RESULTS: In multivariate analysis, ED location predicted improved survival to discharge (OR 0.74, 95%CI [0.67-0.82]). ED CAs had higher survival to discharge rates (ED 22.2, ICU 15.5, Tele 19.8, Floor 10.8, p<0.0001), better cerebral performance category scores (ED 1.59, ICU 1.73, Tele 1.96, Floor 1.69, p<0.0001), and shorter post-event length of stays (ED 8.6, ICU 17.5, Tele 16.5, Floor 14.2 days, p<0.0001) than other locations. Recurrent ED CAs were less likely to survive to discharge (10.1% vs. 24.6%, p<0.0001) than primary events. Trauma related ED CAs had a lower survival to discharge rate (7.5% vs. 23.8%, p<0.0001), were less likely to be caused by an arrhythmia (23.6% vs. 32.5%, p<0.0008), and more likely to be preceded by hypotension or shock (41.6% vs. 29.0%, p<0.0001) than non-trauma ED events. CONCLUSIONS: ED CAs have unique characteristics, and better survival and neurologic outcomes compared to other hospital locations. Primary ED CAs have a better chance of survival to discharge than recurrent events. Traumatic ED CAs have worse outcomes than non-traumatic CA. PMID- 18508186 TI - Interaction of 9,10-anthraquinone with adenine and 2'-deoxyadenosine. AB - Laser flash photolysis has been used for the study of the interaction of 9,10 anthraquinone (AQ) with the DNA base, adenine (A) and its corresponding nucleoside, 2'-deoxyadenosine (dA). This study has provided two very important observations. AQ has been found to support electron transfer in different categories of media, acetonitrile/water on one hand and SDS micelles on other. While in our earlier work 2-methyl 1,4-naphthoquinone was found to undergo a switchover in reactivity (J. Am. Chem. Soc. 126 (2004) 10589-10593). Again A and dA are found to behave differently on account of an extra sugar unit, which not only affects the rate of reaction but the reaction pathway has been found to be modified too. PMID- 18508185 TI - Kinetic analysis of acetylation-dependent Pb1 bromodomain-histone interactions. AB - Stopped-flow fluorescence anisotropy was used to determine the kinetic parameters that define acetylation-dependent bromodomain-histone interactions. Bromodomains are acetyllysine binding motifs found in many chromatin associated proteins. Individual bromodomains were derived from the polybromo-1 protein, which is a subunit of the PBAF chromatin-remodeling complex that has six tandem bromodomains in the amino-terminal region. The average k(on) and k(off) values for the formation of high-affinity complexes are 275 M(-1) s(-1) and 0.41 x 10(-3) s(-1), respectively. The average k(on) and k(off) values for the formation of low affinity complexes are 119 M(-1) s(-1) and 1.42 x 10(-3) s(-1), respectively. Analysis of the on- and off-rates yields acetylation site-dependent equilibrium dissociation constants averaging 1.4 and 12.9 microM for high- and low-affinity complexes, respectively. This work represents the first examination of kinetic mechanisms of acetylation-dependent bromodomain-histone interactions. PMID- 18508187 TI - Fetal-maternal microchimerism in normal parous females and parous female cancer patients. AB - OBJECTIVE: The prevalence of male microchimerism (MC) in parous females, nonparous females, and parous female cancer patients was examined. MATERIALS AND METHODS: DNA extracted from peripheral blood leukocytes and male Y-chromosomal DNA was amplified using a sensitive two-stage polymerase chain reaction technique. Controls prepared by mixing human male and female cell lines demonstrated the sensitivity of the technique to be in the range of 1 male cell per 1 million female cells. RESULTS: Findings of this study showed that the percentage of MC-positive females was highly dependent on the amount of DNA analyzed; 57% of normal parous females who bore at least one son were found to have male cells in their blood when 25 mug DNA or more from the samples was analyzed. This frequency is much higher than previous reports indicating a prevalence of 33% for normal parous females. Analysis of samples obtained from 200 parous female cancer patients revealed an incidence of 34% MC(+); 7.4% of normal nonparous female controls had evidence of MC. CONCLUSION: The long-term persistence of male cells in the maternal circulation could indicate maternal immune tolerance of paternally inherited fetal antigens. This maternal tolerance might be exploited in female patients with malignant disease to deliver immune cellular therapy from their sons. PMID- 18508188 TI - Mid-term results demonstrate salvage high-intensity focused ultrasound (HIFU) as an effective and acceptably morbid salvage treatment option for locally radiorecurrent prostate cancer. AB - BACKGROUND: Local occurrence of prostate cancer (PCa) after external beam radiation (EBRT) may benefit from definitive local therapy. OBJECTIVE: To evaluate the safety and efficacy of salvage high-intensity focal ultrasound (HIFU) in local PCa recurrence after EBRT and to determine prognostic factors for optimal patient selection. DESIGN, SETTING, AND PARTICIPANTS: Between 1995 and 2006, patients with a local PCa recurrence after EBRT were retrospectively included. INTERVENTION: All patients received salvage HIFU with the Ablatherm device. MEASUREMENTS: Prognostic factors (pre-EBRT risk group, androgen deprivation [AD] use, pre-HIFU prostate-specific antigen [PSA], Gleason score and positive biopsy percentage) were studied in univariate and multivariate analyses. Progression was defined as positive biopsy and/or last PSA > nadir + 2 ng/ml and/or adjuvant therapy introduction. All complications were recorded. RESULTS AND LIMITATIONS: Some 194 HIFU sessions for 167 patients were performed. Local cancer control was achieved with negative biopsy results in 122 (73%) patients. The median PSA nadir was 0.19 ng/ml. The mean follow-up period was 18.1 mo (range: 3-121 mo). Seventy-four patients required no hormone therapy. The actuarial 5-yr overall survival rate was 84%. The actuarial 3-yr progression-free survival rate was significantly lower in three circumstances: (1) worsening of the pre-EBRT stage with 53%, 42%, and 25% for low-, intermediate-, and high-risk patients, respectively, (2) increase in the pre-HIFU PSA, and (3) use of AD during PCa management. In multivariate analyses, the risk ratio for intermediate- and high-risk patients were 1.32 and 1.96, respectively. The risk ratio was 2.8 if patients had received AD. No rectal complications were observed. Urinary incontinence accounted for 49.5% of the urinary sphincter implantations required in 11% of patients. This is a retrospective study in which the role of the PSA doubling time and the time until recurrence was not evaluated. CONCLUSIONS: Salvage HIFU is a curative treatment option for local relapse after EBRT with acceptable morbidity. Careful patient selection is imperative depending upon the aforementioned prognostic factors. PMID- 18508189 TI - Editorial comment on: Mid-term results demonstrate salvage high-intensity focused ultrasound (HIFU) as an effective and acceptably morbid salvage treatment option for locally radiorecurrent prostate cancer. PMID- 18508190 TI - Editorial comment on: Mid-term results demonstrate salvage high-intensity focused ultrasound (HIFU) as an effective and acceptably morbid salvage treatment option for locally radiorecurrent prostate cancer. PMID- 18508191 TI - Re: Marco Roscigno, Cesare Cozzarini, Roberto Bertini, et al. Prognostic value of lymph node dissection in patients with muscle-invasive transitional cell carcinoma of the upper urinary tract. Eur Urol 2008;53:794-802. PMID- 18508192 TI - AKR1C2 and AKR1C3 mediated prostaglandin D2 metabolism augments the PI3K/Akt proliferative signaling pathway in human prostate cancer cells. AB - Members of the aldo-keto reductase (AKR) superfamily have been implicated in prostaglandin (PG) metabolism and prostate cancer. AKR1C3 possesses 11 ketoprostaglandin reductase activity and is capable of converting PGD2 to 9alpha, 11beta-PGF2alpha, whereas AKR1C2-mediated PG metabolism remains unclear. The accumulation of PGF2alpha may generate proliferative signals to promote prostate cell growth. Levels of AKR1C2 and AKR1C3 expression are elevated in localized and advanced prostate cancer. To study the significance of AKR1C2- and AKR1C3 mediated PGD2 conversion in human prostate cell proliferation, we stably transfected androgen insensitive human prostate cancer PC-3 cells with AKR1C2 or AKR1C3 cDNA. PC-3 cells overexpressing AKR1C2 and AKR1C3 had elevated cell proliferation in response to PGD2 stimulation as compared to mock transfectants. Overexpression of AKR1C2 or AKR1C3 did not alter levels of PGF receptor (FP) expression. Inclusion of an FP antagonist (AL8810) significantly suppressed PGD2 stimulated PC-3 cell proliferation in these stable transfectants. In addition, PGD2 significantly elevated levels of total Akt protein expression and Akt Ser473 phosphorylation in AKR1C2 and AKR1C3 stable transfectants; and inclusion of a phosphatidylinositol 3-kinase (PI3K) chemical inhibitor (LY294002) attenuated PGD2-stimulated cell proliferation in these transfectants. Our results suggested that both AKR1C2 and AKR1C3 mediate similar PGD2 conversion toward the accumulation of proliferative signals through FP and PI3K/Akt signaling pathways to promote prostate cell proliferation. PMID- 18508193 TI - Think globally: act locally. New insights into the local regulation of thyroid hormone availability challenge long accepted dogmas. AB - Recent evidence derived from transgenic mouse models and findings in humans with mutations affecting thyroid hormone (TH) metabolism have convincingly supported a model of TH signalling in which regulated local adjustment of active TH concentrations is far more important than circulating plasma hormone levels. Although this theory was put forward several years ago and has been supported by significant, but inherently indirect evidence, recent insights from targeted deletion of the genes encoding deiodinase (Dio) isozymes have revived this model and greatly increased our understanding of TH metabolism. However, gene targeting proved to be a double edged sword, since the overall model was supported, but several predictions are apparently not consistent with the new experimental evidence. Human genetics further provided additional exciting data on the physiological role of Dio isozymes that need to be incorporated into any model of TH biology. The recent identification of mutations in the T3 plasma membrane transporter MCT8 has sparked new interest in the role of TH in brain function, since affected patients suffered from psychomotor retardation. Moreover, selenium (Se) and TH physiology have finally been unequivocally connected by newly identified inherited defects in a gene involved in selenoprotein biosynthesis. Finally, a link between Dio expression and energy metabolism has been delineated in mice that may hold great promise for the management of the adiposity pandemic. PMID- 18508194 TI - Adsorption of lead(II) ions onto 8-hydroxy quinoline-immobilized bentonite. AB - In this study, the immobilization of 8-hydroxy quinoline onto bentonite was carried out and it was then used to investigate the adsorption behavior of lead(II) ions from aqueous solutions. The changes of the parameters of pH, contact time, initial lead(II) ions concentration and temperature were tested in the adsorption experiments. The XRD, FTIR, elemental and thermal analyses were done to observe the immobilization of 8-hydroxy quinoline onto natural bentonite. The adsorption was well described by the Langmuir adsorption isotherm model at all studied temperatures. The maximum adsorption capacity was 142.94mgg(-1) from the Langmuir isotherm model at 50 degrees C. The thermodynamic parameters implied that the adsorption process is spontaneous and endothermic. The kinetic data indicate that the adsorption fits well with the pseudo-second-order kinetic model. 8-Hydroxy quinoline-immobilized bentonite can be used as well respective adsorbent for the removal of the heavy metal pollutants according to the results. PMID- 18508195 TI - Effect of synthesis parameters on the compressive strength of low-calcium ferronickel slag inorganic polymers. AB - The wide range of physical and chemical properties of inorganic polymers, also known as geopolymers, commonly formed by alkali activation of aluminosilicates, makes these materials useful for a variety of applications. In the present experimental study inorganic polymers are synthesised from low-Ca electric arc ferronickel slag. The effect of experimental conditions on the compressive strength of the final products is assessed. A number of techniques, namely XRD, FTIR and TG-DTG were used to identify new phases and subsequently elucidate to some degree the mechanisms involved. Finally, the paper discusses briefly the potential of inorganic polymer technology as a feasible option for the utilisation of certain potentially hazardous mining and metallurgical wastes towards an increased sustainability of the wider minerals sector. PMID- 18508196 TI - Removal of ammonium ion from produced waters in petroleum offshore exploitation by a batch single-stage electrolytic process. AB - This work describes a batch single-stage electrochemical process to remove quantitatively the ammonium ion from produced waters from petroleum exploration of the Campos' Basin, seeking to fulfil the directories of the National Brazilian Environmental Council. The anode was made out of titanium covered by a layer of RuO(2)+TiO(2) oxides (Dimensionally Stable Anode), whereas the cathode was made out of pure titanium. Anodic and cathodic compartments were separated by a membrane. The applied current varied from 0.3 to 1.5A. As the current increased NH(4)(+) removal was faster and pH was rapidly decreased to 3. The pH of the anodic compartment increased to approximately 10. When the current was 0.92 A chlorine evolution was observed after 40 min or only 15 min when that current was 1.50 A. In this voltage a deposit containing alkali-earth metal hydroxides/sulphates was formed on the membrane surface of the cathode side, thus suggesting a diffusion process from the anodic to the cathodic compartment. The maximum current applied to the cell must not exceed approximately 0.70 A in order to avoid chlorine evolution. Ammonia removal was over 99.9 wt% at 0.68 A in about 75 min. PMID- 18508197 TI - Kinetic and thermodynamic aspects of Cu(II) and Cr(III) removal from aqueous solutions using rose waste biomass. AB - Distillation waste of rose petals was used to remove Cu(II) and Cr(III) from aqueous solutions. The results demonstrated the dependency of metal sorption on pH, sorbent dose, sorbent size, initial bulk concentration, time and temperature. A dosage of 1g/L of rose waste biomass was found to be effective for maximum uptake of Cu(II) and Cr(III). Optimum sorption temperature and pH for Cu(II) and Cr(III) were 303+/-1K and 5, respectively. The Freundlich regression model and pseudo-second-order kinetic model were resulted in high correlation coefficients and described well the sorption of Cu(II) and Cr(III) on rose waste biomass. At equilibrium q(max) (mg/g) of Cu(II) and Cr(III) was 55.79 and 67.34, respectively. The free energy change (DeltaG degrees ) for Cu(II) and Cr(III) sorption process was found to be -0.829 kJ/mol and -1.85 kJ/mol, respectively, which indicates the spontaneous nature of sorption process. Other thermodynamic parameters such as entropy change (DeltaS degrees ), enthalpy (DeltaH degrees )and activation energy (DeltaE) were found to be 0.604 J mol(-1)K(-1), -186.95 kJ/mol and 68.53 kJ/mol, respectively for Cu(II) and 0.397 J mol(-1)K(-1), 119.79 kJ/mol and 114.45 kJ/mol, respectively for Cr(III). The main novelty of this work was the determination of shortest possible sorption time for Cu(II) and Cr(III) in comparison to earlier studies. Almost over 98% of Cu(II) and Cr(III) were removed in only first 20 min at an initial concentration of 100 mg/L. PMID- 18508198 TI - Stimulation of myofascial trigger points with ultrasound induces segmental antinociceptive effects: a randomized controlled study. AB - Musculoskeletal pain affects a significant proportion of the general population. The myofascial trigger point is recognized as a key factor in the pathophysiology of musculoskeletal pain. Ultrasound is commonly employed in the treatment and management of soft tissue pain and, in this study, we set out to investigate the segmental antinociceptive effect of ultrasound. Subjects (n=50) with identifiable myofascial trigger points in the supraspinatus, infraspinatus and gluteus medius muscles were selected from an outpatient rehabilitation clinic and randomly assigned to test or control groups. Test subjects received a therapeutic dose of ultrasound to the right supraspinatus trigger point while control groups received a sham (null) exposure. Baseline pain pressure threshold (PPT) readings were recorded at the ipsilateral infraspinatus and gluteus medius trigger-point sites prior to ultrasound exposure. The infraspinatus point was chosen due to its segmental neurologic link with the supraspinatus point; the gluteus medius acted as a segmental control point. Following the ultrasound intervention, PPT readings were recorded at 1, 3, 5, 10 and 15 min intervals at both infraspinatus and gluteus medius trigger points; the difference between infraspinatus and gluteus medius PPT values, PPT seg, represents the segmental influence on the PPT. The ultrasound test group demonstrated statistically significant increases in PPT seg (decreased infraspinatus sensitivity) at 1, 3 and 5 min, when compared with PPT seg in the sham ultrasound group. These results establish that low-dose ultrasound evokes short-term segmental antinociceptive effects on trigger points which may have applications in the management of musculoskeletal pain. PMID- 18508199 TI - Restoration of weak phase-contrast images recorded with a high degree of defocus: the "twin image" problem associated with CTF correction. AB - Relatively large values of objective-lens defocus must normally be used to produce detectable levels of image contrast for unstained biological specimens, which are generally weak phase objects. As a result, a subsequent restoration operation must be used to correct for oscillations in the contrast transfer function (CTF) at higher resolution. Currently used methods of CTF correction assume the ideal case in which Friedel mates in the scattered wave have contributed pairs of Fourier components that overlap with one another in the image plane. This "ideal" situation may be only poorly satisfied, or not satisfied at all, as the particle size gets smaller, the defocus value gets larger, and the resolution gets higher. We have therefore investigated whether currently used methods of CTF correction are also effective in restoring the single-sideband image information that becomes displaced (delocalized) by half (or more) the diameter of a particle of finite size. Computer simulations are used to show that restoration either by "phase flipping" or by multiplying by the CTF recovers only about half of the delocalized information. The other half of the delocalized information goes into a doubly defocused "twin" image of the type produced during optical reconstruction of an in-line hologram. Restoration with a Wiener filter is effective in recovering the delocalized information only when the signal-to-noise ratio (S/N) is orders of magnitude higher than that which exists in low-dose images of biological specimens, in which case the Wiener filter approaches division by the CTF (i.e. the formal inverse). For realistic values of the S/N, however, the "twin image" problem seen with a Wiener filter is very similar to that seen when either phase flipping or multiplying by the CTF is used for restoration. The results of these simulations suggest that CTF correction is a poor alternative to using a Zernike-type phase plate when imaging biological specimens, in which case the images can be recorded in a close-to focus condition, and delocalization of high-resolution information is thus minimized. PMID- 18508200 TI - The impact of recent legislation on paediatric fireworks injuries in the Newcastle upon Tyne region. AB - Despite the changes to the UK fireworks laws and considerable efforts in prevention, children are still being injured by fireworks. The UK is one of many countries that have altered their firework laws in recent years. We reviewed 54 firework-injured children over the last 10 years and assessed the impact of the two recent UK law changes. Our study outlines past British firework legislation and reviews the literature. In November 1996, there were three deaths in England, Wales and Scotland due to fireworks. The British Government introduced the Fireworks (Safety) Regulations of 1996/1997, primarily banning banger fireworks (known as bangers). We have not seen banger injuries in Newcastle since then. The Fireworks Act 2003 and the Fireworks Regulations 2004 limited the sale of fireworks to the 3 weeks surrounding bonfire night, and banned under 18s from purchasing or possessing fireworks. In our series, we noticed that, in 2004, 83% of children's firework injuries happened in the 3 weeks surrounding Bonfire Night. We conclude that legislation has had an impact, but stricter enforcement of the existing laws and further education of children and the general public into the dangers of fireworks is needed, as children are still being injured. PMID- 18508201 TI - Cellular recovery from electroporation using synchronisation modulation as a rescue model for electrically injured cells. AB - Electroporation of the plasma membrane resulting in a decrement in transmembrane potential is offered as a model in the study of the rescuing effects of the synchronisation modulation technique by electrically activating sodium potassium adenosine triphosphatase. Living cells were first electrically damaged by a pulsed intensive electric field, resulting in cell membrane electroporation, ion leakages and membrane potential depolarisation. Their recovery rate in natural conditions was compared with that of cells in a synchronisation modulation electric field. Fluorescence readings were taken using confocal microscopy and a potentiometric dye. Significantly more rapid recovery was observed after synchronisation modulation, with cell membranes actually polarised to levels higher than the original resting potential, a feature never seen in naturally recovering cells. PMID- 18508202 TI - Glitazones and skin aging: may they stop the ticking clock? PMID- 18508203 TI - Efficacy and factors affecting outcome of gemcitabine concurrent chemoradiotherapy in patients with locally advanced pancreatic cancer. AB - PURPOSE: To evaluate the efficacy and prognostic factors of gemcitabine (GEM) concurrent chemoradiotherapy (CCRT) in patients with locally advanced pancreatic cancer. METHODS AND MATERIALS: Between January 2002 and December 2005, 55 patients with locally advanced pancreatic cancer treated with GEM (400 mg/m(2)/wk) concurrently with radiotherapy (median dose, 50.4 Gy; range, 26-61.2) at Taipei Veterans General Hospital were enrolled. GEM (1,000 mg/m(2)) was continued after CCRT as maintenance therapy once weekly for 3 weeks and repeated every 4 weeks. The response, survival, toxicity, and prognostic factors were evaluated. RESULTS: With a median follow-up of 10.8 months, the 1- and 2-year survival rate was 52% and 19%, respectively. The median overall survival (OS) and median time to progression (TTP) was 12.4 and 5.9 months, respectively. The response rate was 42% (2 complete responses and 21 partial responses). The major Grade 3-4 toxicities were neutropenia (22%) and anorexia (19%). The median OS and TTP was 15.8 and 9.5 months in the GEM CCRT responders compared with 7.5 and 3.5 months in the nonresponders, respectively (both p < 0.001). The responders had a better Karnofsky performance status (KPS) (86 +/- 2 vs. 77 +/- 2, p = 0.002) and had received a greater GEM dose intensity (347 +/- 13 mg/m(2)/wk vs. 296 +/- 15 mg/m(2)/wk, p = 0.02) than the nonresponders. KPS and serum carbohydrate antigen 19-9 were the most significant prognostic factors of OS and TTP. CONCLUSION: The results of our study have shown that GEM CCRT is effective and tolerable for patients with locally advanced pancreatic cancer. The KPS and GEM dose correlated with response. Also, the KPS and CA 19-9 level were the most important factors affecting OS and TTP. PMID- 18508204 TI - Beneficial effect of Embelia ribes ethanolic extract on blood pressure and glycosylated hemoglobin in streptozotocin-induced diabetes in rats. AB - Embelia ribes (common name, Vidanga) is extensively used in traditional system of medicine for treatment of various disorders. It is described in Ayurveda, as a powerful anthelmintic, antifertility and antihyperlipidemic agent. The present study was undertaken to investigate modulatory effect of 6 weeks' chronic oral administration of E. ribes ethanolic extract on diabetes mellitus induced by a diabetogen, streptozotocin (STZ) with special reference to changes in glucose levels, glycated haemoglobin status and cardiac toxicity. STZ treatment (40 mg/kg iv) resulted in significant increase in blood glucose levels, glycated haemoglobin levels, heart rate (HR) and systolic blood pressure (SBP). Oral administration of E. ribes ethanolic extract in dose of 100 mg/kg and 200 mg/kg significantly reduced the levels of blood glucose, glycated haemoglobin, heart rate (HR) and systolic blood pressure (SBP) in animals when compared with diabetic rats. PMID- 18508205 TI - Alpha-glucosidase inhibitor from Chinese aloes. AB - Activity-guided fractionation of the methanol extract of Chinese aloes led to the isolation of aloeresin A, which demonstrated significant dose dependent alpha glucosidase inhibitory activities, with IC(50) values of 11.94 and 2.16 mM, against rat intestinal sucrase and maltase, respectively. PMID- 18508206 TI - [Psychological impact of miscarriage: a review]. AB - Miscarriage is a frequent but painful event for women. This review aims to underscore psychological morbidity after miscarriage and some characteristics liable to influence women's experience. Then, the last part of the article concerns importance of psychological counselling and different counselling interventions, which have been tested. This review has for objective to provide informations about psychological experience of miscarriage for practitioners and medical teams who take care of women. PMID- 18508207 TI - A Phase 3 placebo-controlled, double-blind, multi-site trial of the alpha-2 adrenergic agonist, lofexidine, for opioid withdrawal. AB - CONTEXT: Lofexidine is an alpha-2-adrenergic receptor agonist that is approved in the United Kingdom for the treatment of opioid withdrawal symptoms. Lofexidine has been reported to have more significant effects on decreasing opioid withdrawal symptoms with less hypotension than clonidine. OBJECTIVE: To demonstrate that lofexidine is well tolerated and effective in the alleviation of observationally defined opioid withdrawal symptoms in opioid dependent individuals undergoing medically supervised opioid detoxification as compared to placebo. DESIGN: An inpatient, Phase 3, placebo-controlled, double-blind, randomized multi-site trial with three phases: (1) opioid agonist stabilization phase (days 1-3), (2) detoxification/medication or placebo phase (days 4-8), and (3) post detoxification/medication phase (days 9-11). SUBJECTS: Sixty-eight opioid dependent subjects were enrolled at three sites with 35 randomized to lofexidine and 33 to placebo. MAIN OUTCOME MEASURE: Modified Himmelsbach Opiate Withdrawal Scale (MHOWS) on study day 5 (second opioid detoxification treatment day). RESULTS: Due to significant findings, the study was terminated early. On the study day 5 MHOWS, subjects treated with lofexidine had significantly lower scores (equating to fewer/less severe withdrawal symptoms) than placebo subjects (least squares means 19.5+/-2.1 versus 30.9+/-2.7; p=0.0019). Lofexidine subjects had significantly better retention in treatment than placebo subjects (38.2% versus 15.2%; Log rank test p=0.01). CONCLUSIONS: Lofexidine is well tolerated and more efficacious than placebo for reducing opioid withdrawal symptoms in inpatients undergoing medically supervised opioid detoxification. PMID- 18508208 TI - Efficacy of natural propolis extract in the control of American Foulbrood. AB - Paenibacillus larvae is the causative agent of American Foulbrood (AFB), a severe disease that affects larvae of the honeybees. Due to the serious effects associated with AFB and the problems related to the use of antibiotics, it is necessary to develop alternative strategies for the control of the disease. The aim of the present work was to evaluate the effect of a propolis ethanolic extract (PEE) against P. larvae and its potential for the control of AFB. In vitro activity of PEE against P. larvae isolates was evaluated by the disk diffusion method and the minimum inhibitory concentration (MIC) was determined. Toxicity for honeybees was evaluated by oral administration of PEE and its lethal concentration was assessed. Lastly, colonies from an apiary with episodes of AFB on previous years were divided into different groups and treated with sugar syrup supplemented with PEE by aspersion (group one), sugar syrup by aspersion (group two), fed with sugar syrup supplemented with PEE (group three) and fed with sugar syrup only (group four). All isolates were sensitive to PEE and the MIC median was 0.52% (range 0.32-0.64). PEE was not toxic for bees at least at 50%. Field assays showed that 21 and 42 days after the application of the treatments, the number of P. larvae spores/g of honey was significantly lower in colonies treated with PEE compared to the colonies that were not treated with PEE. To our knowledge, this is the first report about the use of propolis for the treatment of beehives affected with P. larvae spores. PMID- 18508209 TI - Cellular responses induced by silver nanoparticles: In vitro studies. AB - A systematic study on the in vitro interactions of 7-20 nm spherical silver nanoparticles (SNP) with HT-1080 and A431 cells was undertaken as a part of an on going program in our laboratory to develop a topical antimicrobial agent for the treatment of burn wound infections. Upon exposure to SNP (up to 6.25 microg/mL), morphology of both the cell types remained unaltered. However, at higher concentrations (6.25-50 microg/mL) cells became less polyhedral, more fusiform, shrunken and rounded. IC(50) values for HT-1080 and A431 as revealed by XTT assay were 10.6 and 11.6 microg/mL, respectively. When the cells were challenged with approximately 1/2 IC(50) concentration of SNP (6.25 microg/mL), clear signs of oxidative stress, i.e. decreased GSH ( approximately 2.5-folds in HT-1080, approximately 2-folds in A431) and SOD ( approximately 1.6-folds in HT-1080, 3 folds in A431) as well as increased lipid peroxidation ( approximately 2.5-folds in HT-1080, approximately 2-folds in A431) were seen. Changes in the levels of catalase and GPx in A431 cells were statistically insignificant in both cell types. DNA fragmentation in SNP-exposed cells suggested apoptosis. When the apoptotic thresholds of SNP were monitored with caspase-3 assay the concentrations required for the onset of apoptosis were found to be much lower (0.78 microg/mL in HT-1080, 1.56 microg/mL in A431) than the necrotic concentration (12.5 microg/mL in both cell types). These results can be used to define a safe range of SNP for the intended application as a topical antimicrobial agent after appropriate in vivo studies. PMID- 18508210 TI - The validity and reliability of the Malaysian Osteoporosis Knowledge Tool in postmenopausal women. AB - OBJECTIVES: To design a valid and reliable questionnaire, the Malaysian Osteoporosis Knowledge Tool (MOKT), to assess the knowledge of postmenopausal osteoporotic women in Malaysia, on osteoporosis and its treatment. METHODS: A 40 item instrument was designed and tested in this study. The scores ranged from 1 to 40, which were then converted into percentage. This was administered to 88 postmenopausal osteoporotic women who were taking alendronate (patient group) and 43 pharmacists (professional group). The MOKT was administered again to the patient group after 1 month. RESULTS: Flesch reading ease was 57, which is satisfactory, while the mean difficulty factor+/-S.D. was 0.73+/-0.25, indicating that the MOKT is moderately easy. Internal consistency of the instrument was good with Cronbach's alpha=0.82. The test and retest scores showed no significant difference for 38 out of the 40 items, indicating that the questionnaire has achieved a stable reliability. Level of knowledge on osteoporosis and its treatment was good with a mean score+/-S.D. of 69.0+/-13.9 and 81.6+/-9.5 in the patient and professional group, respectively. The difference in knowledge scores between the two groups was statistically significant (p<0.001). CONCLUSIONS: In conclusion, the 40-item questionnaire (MOKT) is a reliable and valid instrument for measuring knowledge on osteoporosis in the Malaysian setting. Therefore, it can be used to identify individuals in need of educational interventions and to assess the effectiveness of education efforts as part of osteoporosis management. PMID- 18508211 TI - Solid lipid nanoparticles for retinal gene therapy: transfection and intracellular trafficking in RPE cells. AB - Retinal pigment epithelial (RPE) cells are usually employed to study DNA systems for diseases related to problems in the retina. Solid lipid nanoparticles (SLNs) have been shown to be useful non-viral vectors for gene therapy. The objective of this work was to evaluate the transfection capacity of SLNs in the human retinal pigment epithelial established cell line (ARPE-19) in order to elucidate the potential application of this vector in the treatment of retinal diseases. Results showed a lower transfection level of SLNs in ARPE-19 cells than in HEK293 (2.5% vs. 14.9% EGFP positive cells at 72h post-transfection). Trafficking studies revealed a delay in cell uptake of the vectors in ARPE-19 cells. Differences in internalization process into the two cell lines studied explain, in part, the difference in the gene expression. The clathrin-mediated endocytosis in ARPE-19 cells directs the solid lipid nanoparticles to lysosomes; moreover, the low division rate of this cell line hampers the entrance of DNA into the nucleus. The knowledge of intracellular trafficking is very useful in order to design more efficient vectors taking into account the characteristics of the specific cell line to be transfected. PMID- 18508212 TI - Pharmacokinetics and pharmacodynamics of chlorambucil delivered in parenteral emulsion. AB - The aim was to assess the pharmacokinetics and anticancer activity of chlorambucil (CHL) incorporated in a parenteral emulsion (PE). A chlorambucil loaded PE was prepared by a high energy ultrasonication method. Soybean oil was chosen as a triglyceride oil core and egg phosphatidylcholine as an emulsifier in the formulation. The particle size distribution and zeta potential were measured using Zetasizer. The results showed that the average encapsulation efficiency of chlorambucil-loaded parenteral emulsion (CHL-PE) was 98.6+/-3.2% with a particle size of 182.7+/-0.8 nm, and a zeta-potential of -37.2+/-1.1 mV. Osmolality and pH of the formulation were 305.6+/-2.3 mOsm/kg and 7.4, respectively. The chlorambucil was stable in the PE for at least 6 months stored at 4-8 degrees C. The pharmacokinetics, tissue distribution, and anticancer activity of CHL-PE and chlorambucil solution were studied after intravenous administration to C57 BL/6 male mice. CHL-PE exhibited a significantly greater AUC 0-infinity (32.4+/-0.1 microg/ml h vs. 16.9+/-0.1 microg/ml h), mean residence time (MRT) (1.32+/-0.01 h vs. 0.30+/-0.01 h), volume of distribution (409+/-15 ml/kg vs. 180+/-7 ml/kg) and elimination half-life (1.83+/-0.1h vs. 0.27+/-0.02 h) (all P<0.01), and a significantly reduced plasma clearance (309+/-16 ml/(h kg) vs. 591+/-4 ml/(h kg), P<0.01) compared to the CHL. In addition CHL-PE treatment caused significantly greater tumour growth suppression rate (% T/C) of the colon-38 adenocarcinoma in the mouse compared to CHL treatment (% T/C, 75+/-3.4% vs. 49+/-7.4%, P<0.01). These results suggest that CHL-PE could be an effective parenteral carrier for chlorambucil delivery in cancer treatment. PMID- 18508213 TI - Age-related hearing loss in the Fischer 344/NHsd rat substrain. AB - Studies of the F344 rat have shown a variety of age-related auditory anatomy and physiology changes. The current study was undertaken to clarify the ARHL in the F344 rat, by examining the auditory pathway of the F344/NHsd substrain that is distributed by Harlan Laboratories for research in the United States. The F344/NHsd rat begins to lose its hearing at about 12 months, and by 24 months, there are 50-60 dB auditory brainstem response threshold shifts at 20 and 40 kHz and 20 dB losses at 5-10 kHz. Distortion product otoacoustic emissions (DPOAE) amplitudes at 1.8-12 kHz stimuli were depressed in the older (18-24 months) rats. Amplitude input-output functions of the compound action potential (CAP) were also depressed across frequency. The endocochlear potential (EP) was 90-100 mV in the 3 month old rats. All but one of the 24 month old rats' EPs were in the +75-85 mV range. Tympanometry revealed no differences in middle ear function between the young and older rats. Collectively, these findings suggest damage to the outer hair cells, but anatomical examination of the outer hair cells revealed a relative lack of cell loss compared to the magnitude of the hearing and DPOAE loss. PMID- 18508214 TI - Cytotoxic effects of compounds from Iris tectorum on human cancer cell lines. AB - In the course of searching for novel cytotoxic compounds which can be used in chemotherapy, several Traditional Chinese Medicines (TCM) have been screened by bioassay-guided fractionation and isolation. An extract of rhizomes of Iris tectorum Maxim., a TCM used to treat cancer, exhibited highest potency and led to the isolation of two flavonoids, 7-O-methylaromadendrin and tectorigenin, and four iridal-type triterpenes, iritectols A and B, isoiridogermanal and iridobelamal A. The cytotoxicities of the isolated compounds against four human cancer cell lines were evaluated by the SRB assay. Iritectol B, isoiridogermanal and iridobelamal A showed similar cytotoxicity with IG(50) around 11 microM and 23 microM against MCF-7 and C32 cell lines, respectively. Cell cycle-specific inhibition and apoptosis induced by the isolated compounds were determined using flow cytometry with two sets of co-labelling systems: annexin V-FITC/propidium iodide and fluorescein diacetate/propidium iodide. Iritectol B demonstrated dose dependent apoptotic effect against COR-L23 cells with an apoptotic rate of 33% at 100 microM. Tectorigenin (an analogue of genistein) showed cell cycle specific inhibition and arrested cells at G(2)/M phase up to 400 microM, but did not demonstrate apoptotic effect against COR-L23 cells up to 1 mM. The overall activities of isolated compounds observed in the present study support the traditional use of Iris tectorum Maxim. in the treatment of cancer. PMID- 18508215 TI - Chemical markers from the peracid oxidation of isosafrole. AB - In this work, isomers of 2,4-dimethyl-3,5-bis(3,4 methylenedioxyphenyl)tetrahydrofuran (11) are presented as chemical markers formed during the peracid oxidation of isosafrole. The stereochemical configurations of the major and next most abundant diastereoisomer are presented. Also described is the detection of isomers of (11) in samples from a clandestine laboratory uncovered in South Australia in February 2004. PMID- 18508216 TI - High-resolution intracranial MRA at 7T using autocalibrating parallel imaging: initial experience in vascular disease patients. AB - PURPOSE: Greater spatial resolution in intracranial three-dimensional time-of flight (TOF) magnetic resonance angiography (MRA) is possible at higher field strengths, due to the increased contrast-to-noise ratio (CNR) from the higher signal-to-noise ratio and the improved background suppression. However, at very high fields, spatial resolution is limited in practice by the acquisition time required for sequential phase encoding. In this study, we applied parallel imaging to 7T TOF MRA studies of normal volunteers and patients with vascular disease, in order to obtain very high resolution (0.12 mm(3)) images within a reasonable scan time. MATERIALS AND METHODS: Custom parallel imaging acquisition and reconstruction methods were developed for 7T MRA, based on generalized autocalibrating partially parallel acquisition (GRAPPA). The techniques were compared and applied to studies of seven normal volunteers and three patients with cerebrovascular disease. RESULTS: The technique produced high resolution studies free from discernible reconstruction artifacts in all subjects and provided excellent depiction of vascular pathology in patients. CONCLUSIONS: 7T TOF MRA with parallel imaging is a valuable noninvasive angiographic technique that can attain very high spatial resolution. PMID- 18508217 TI - Exploring the feasibility of simultaneous electroencephalography/functional magnetic resonance imaging at 7 T. AB - The increased blood oxygenation level-dependent contrast available at high field makes the implementation of combined EEG/fMRI experiments at 7 T highly worthwhile from the point of view of fMRI data quality, but the higher field poses greater technical challenges for achieving good quality EEG data. A study of the feasibility of recording EEG signals from human subjects at 7 T using a commercially available, MR-compatible EEG system has therefore been carried out. This involved systematic measurement of the sources of noise in EEG recordings made in the 7 T scanner and measurement of RF heating effects on a gel phantom in the presence of a 32-electrode EEG cap. Having found no significant safety concerns and identified a set-up (involving switching off the magnet's cryo cooler pumps and mounting the EEG amplifier on a cantilever) that limited scanner induced noise, combined EEG/fMRI experiments employing visual stimulation were then successfully carried out on two human subjects. With the use of beamformer based analysis of the EEG data, driven responses and alpha-band, event-related desynchronisation were identified in both subjects. PMID- 18508218 TI - Multivariate analysis of diffusion tensor imaging data improves the detection of microstructural damage in young professional boxers. AB - In this study, we present two different methods of multivariate analysis of voxel based diffusion tensor imaging (DTI) data, using as an example data derived from 59 professional boxers and 12 age-matched controls. Conventional univariate analysis ignores much of the diffusion information contained in the tensor. Our first multivariate method uses the Hotelling's T(2) statistic and the second uses linear discriminant analysis to generate the linear discriminant function at each voxel to form a separability metric. Both multivariate methods confirm the findings from the individual metrics of large-scale changes in the bilateral inferior temporal gyri of the boxers, but they also reveal greater sensitivity as well as identifying major subcortical changes that had not been evident in the univariate analyses. Linear discriminant analysis has the added strength of providing a quantitative measure of the relative contribution of each metric to any differences between the two subject groups. This novel adaptation of statistical and mathematical techniques to neuroimaging analysis is important for two reasons. Clinically, it develops the findings of a previous mild head injury study, and, methodologically, it could equally well be applied to multivariate studies of other pathologies. PMID- 18508219 TI - Multivariate analysis of fMRI time series: classification and regression of brain responses using machine learning. AB - Machine learning and pattern recognition techniques are being increasingly employed in functional magnetic resonance imaging (fMRI) data analysis. By taking into account the full spatial pattern of brain activity measured simultaneously at many locations, these methods allow detecting subtle, non-strictly localized effects that may remain invisible to the conventional analysis with univariate statistical methods. In typical fMRI applications, pattern recognition algorithms "learn" a functional relationship between brain response patterns and a perceptual, cognitive or behavioral state of a subject expressed in terms of a label, which may assume discrete (classification) or continuous (regression) values. This learned functional relationship is then used to predict the unseen labels from a new data set ("brain reading"). In this article, we describe the mathematical foundations of machine learning applications in fMRI. We focus on two methods, support vector machines and relevance vector machines, which are respectively suited for the classification and regression of fMRI patterns. Furthermore, by means of several examples and applications, we illustrate and discuss the methodological challenges of using machine learning algorithms in the context of fMRI data analysis. PMID- 18508220 TI - Spatial heterogeneity in the muscle functional MRI signal intensity time course: effect of exercise intensity. AB - It has previously been observed that during isometric dorsiflexion exercise, the time course of T2-weighted signal intensity (SI) changes is spatially heterogeneous. The purpose of this study was to test the hypothesis that this spatial heterogeneity would increase at higher contraction intensities. Eight subjects performed 90-s isometric dorsiflexion contractions at 30% and 60% of maximum voluntary contraction (MVC) while T2-weighted (repetition time/echo time=4000/35 ms) images were acquired. SI was measured before, during and after the contractions in regions of interest (ROIs) in the extensor digitorum longus (EDL) muscle and the deep and superficial compartments of the tibialis anterior (D-TA and S-TA, respectively). For all ROIs at 30% MVC, SI changes were similar. The maximum postcontraction SI was greater than the SI during exercise. At 60% MVC, SI changes during contraction were greater in the S-TA than in the D-TA and EDL. For the EDL and D-TA, the maximum postcontraction SI was greater than those during exercise. For the S-TA, the maximum postcontraction change was greater than the changes at t=8, 20 and 56 s but not the end-exercise value. We conclude that spatial heterogeneity increases during more intense dorsiflexion contractions, possibly reflecting regional differences in perfusion or neural activation of the muscle. PMID- 18508221 TI - Dynamic contrast-enhanced MRI and MR diffusion imaging to distinguish between glandular and stromal prostatic tissues. AB - PURPOSE: To compare peak enhancement (PE), determined from dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI) and the magnetic resonance (MR) directionally-averaged apparent diffusion coefficient () in glandular versus stromal prostatic tissues and, with this comparison, to infer if the hypothesis that gadolinium-DTPA (Gd-DTPA) does not enter healthy glands or ducts is plausible. MATERIALS AND METHODS: MRI, MR spectroscopic imaging, DCE MRI and MR diffusion were evaluated in 17 untreated subjects with suspected or proven prostate cancer. PE and were compared in glandular-ductal tissues [normal peripheral zone and glandular benign prostatic hyperplasia (BPH)] and stromal-low ductal tissues (central gland/mixed BPH and stromal BPH). RESULTS: The glandular ductal tissues had lower PE [125+/-6.4 (% baseline)] and higher [1.57+/-0.15 (s/10(-3) mm2)] than the stromal-low ductal tissues [PE=132+/-5.5 (% baseline) (P< .0008), =1.18+/-0.20 (s/10(-3) mm2) (P< 1 x 10(-8))]. A statistical model based upon stepwise regression was generated and completely separated the tissue types: ductal Measure = 448+669 x (s/10(-3) mm2)-10.7 x PE (1/%), R2=1.0 and P<8 x 10(-10). CONCLUSIONS: The very different MR results in the glandular-ductal versus stromal-low ductal tissues suggest that these tissues have different underlying structure. These results support the hypothesis that Gd-DTPA does not enter healthy prostatic glands or ducts. This may explain the higher PE and lower that previously have been reported in prostate cancer versus healthy tissue. PMID- 18508222 TI - Development and validation of an inductively coupled plasma mass spectrometry method with optimized microwave-assisted sample digestion for the determination of platinum at ultratrace levels in plasma and ultrafiltrate plasma. AB - A highly sensitive inductively coupled plasma mass spectrometry (ICP-MS) method with microwave-assisted sample digestion for the determination of total platinum in rat whole and ultrafiltrate plasma was developed and validated. A first step of this study concerned the optimization of the mineralization procedure, in order to obtain good extraction recovery (higher than 90%) and repeatability (less than 6%) and the absence of matrix effect. ICP-MS analysis was then performed using the "hot plasma/protective ion extraction" mode, achieving high sensitivity and very high signal/noise ratio. Iridium was added as internal standard. The method was then submitted to validation, performed according to the FDA Bioanalytical Validation Methods guidelines and to the Eurachem guide. Validation was carried out in terms of limit of detection (LOD), limit of quantitation (LOQ), linearity, precision, accuracy and stability. An instrumental LOQ of 1.9ngL(-1), corresponding to a concentration of 955ngL(-1) in matrix under the adopted conditions, was obtained, allowing the quantitative analysis of Pt ultratraces. Instrumental linearity was verified in the range 1.9-14,000ngL(-1), corresponding to a concentration range from 955ngL(-1) to 6825microgL(-1) in matrix. Accuracy was evaluated by analyzing control samples for both matrices at different concentration levels; a good agreement (<15%) was obtained. Sample stability was tested by analyzing control samples maintained for 4h at room temperature or submitted to three freezing-thawing cycles. Finally, the developed method was applied to the analysis of plasma and ultrafiltrate plasma of rats treated with oxaliplatin-base drug, thus demonstrating its reliability in pharmacokinetic studies. PMID- 18508223 TI - Development of a method for the determination of caffeine anhydrate in various designed intact tablets [correction of tables] by near-infrared spectroscopy: a comparison between reflectance and transmittance technique. AB - Using near-infrared (NIR) spectroscopy, an assay method which is not affected by such elements of tablet design as thickness, shape, embossing and scored line was developed. Tablets containing caffeine anhydrate were prepared by direct compression at various compression force levels using different shaped punches. NIR spectra were obtained from these intact tablets using the reflectance and transmittance techniques. A reference assay was performed by high-performance liquid chromatography (HPLC). Calibration models were generated by the partial least-squares (PLS) regression. Changes in the tablet thickness, shape, embossing and scored line caused NIR spectral changes in different ways, depending on the technique used. As a result, noticeable errors in drug content prediction occurred using calibration models generated according to the conventional method. On the other hand, when the various tablet design elements which caused the NIR spectral changes were included in the model, the prediction of the drug content in the tablets was scarcely affected by those elements when using either of the techniques. A comparison of these techniques resulted in higher predictability under the tablet design variations using the transmittance technique with preferable linearity and accuracy. This is probably attributed to the transmittance spectra which sensitively reflect the differences in tablet thickness or shape as a result of obtaining information inside the tablets. PMID- 18508224 TI - Unusual form of oxacillin resistance in methicillin-resistant Staphylococcus aureus clinical strains. AB - The mechanisms by which there is differential expression of resistance to oxacillin within the populations of a single strain remains to be fully understood. The purpose of this study was to evaluate and characterize 25 GOA48 methicillin-resistant Staphylococcus aureus (MRSA) oxacillin-susceptible mecA positive strains, which were obtained by screening consecutively 832 S. aureus isolates. These 25 isolates (3% of the total strains investigated) were uniformly detected by extending the 24-h oxacillin agar screen plate to 48 h (namely, GOA48 MRSA). Twenty-two isolates tested positive for penicillin-binding protein 2a, whereas the remaining 3 isolates were inconsistently mecA positive. Inconsistent detection of mecA by polymerase chain reaction (PCR) in the mentioned 3 isolates was investigated by colony hybridization using a mecA probe (> or = 80% of colonies hybridized poorly to the probe). A PCR product that amplified the empty SCCmec insertion site (attB), present only if the element was excised, resulted positive in all 3 isolates before oxacillin exposure, whereas integrated elements were positive only for oxacillin-grown isolates. The remaining 22 strains did not reveal excision demonstrating stable mecA. We concluded that resistance to beta lactams in MRSA-positive mecA strains susceptible to oxacillin is associated to an extreme heterogeneous expression of resistance combined in some cases to oxacillin SCCmec excision. PMID- 18508225 TI - The burden of vancomycin-resistant enterococcal infections in US hospitals, 2003 to 2004. AB - Despite significant concern in the health care community regarding vancomycin resistant enterococci (VRE), there are no estimates of the total number of VRE infections that occur each year in US hospitals. Using data from a national survey of hospital discharges and a national antimicrobial resistance surveillance system, we estimated the annual number of US hospitalization with VRE bloodstream, urinary tract, and wound or intra-abdominal infections. Because of the inexact nature of hospital discharge diagnosis coding, we made both a conservative and liberal estimate of hospitalization with VRE infection by using a variety of data sources. For the years 2003 and 2004, we conservatively estimated that there were 20777 and 20931 VRE infections, respectively; for those same years, the liberal estimates were 78330 and 85586, respectively. Because there are such a large number of hospital discharges for which an infection is coded without an organism code, it is likely that the conservative estimate is an underestimate of the true burden. These estimates highlight the importance of controlling VRE and the need to develop improved methods for tracking the burden of such infections. PMID- 18508227 TI - Etiology of childhood diarrhea in the northeast of Brazil: significant emergent diarrheal pathogens. AB - In a study conducted in Joao Pessoa, northeast of Brazil, 2344 Escherichia coli isolated from 290 infants with diarrhea and 290 healthy matched controls were analyzed for virulence traits. Enteroaggregative E. coli (EAEC) was the most prevalent pathogen associated to acute diarrhea. Based on the results of colony blot hybridization, serotyping, and HEp-2 cell adherence assays, strains were separated in categories as typical enteropathogenic E. coli (EPEC) (1.7%), atypical EPEC (a-EPEC) (9.3%), EAEC (25%), enterotoxigenic E. coli (10%), and enteroinvasive E. coli (EIEC) (1.4%). No enterohemorrhagic E. coli strains were isolated. Other enteropathogens were found, including Salmonella (7.9%), Shigella spp. (4.1%), thermophilic Campylobacter spp. (2.4%), Giardia lamblia (9.3%), and Entamoeba histolytica (5.8%). All enteropathogens were associated with diarrhea (P < 0.01). However, the association was lower for EPEC and EIEC (P < 0.03). Different pathogens associated with diarrhea may have been changing in Brazil where EAEC and a-EPEC seem to be the most prevalent pathogens among them. PMID- 18508226 TI - Integrated results of 2 phase 3 studies comparing tigecycline and levofloxacin in community-acquired pneumonia. AB - Tigecycline (TGC), a glycylcycline, has expanded activity against Gram-positive and Gram-negative, anaerobic, and atypical bacteria. Two phase 3 studies were conducted. Hospitalized patients with community-acquired pneumonia (CAP) were randomized to intravenous (IV) TGC (100 mg followed by 50 mg bid) or IV levofloxacin (LEV) (500 mg bid). In 1 study, patients could be switched to oral LEV after at least 3 days intravenously. The coprimary efficacy end points were as follows: clinical response in clinically evaluable (CE) and clinical modified intent-to-treat (c-mITT) populations at test-of-cure (TOC). The secondary end points were as follows: microbiologic efficacy and susceptibility to TGC for CAP bacteria. Safety evaluations were included. Eight hundred ninety-one were patients screened: 846 mITT (TGC 424, LEV 422), 574 CE (TGC 282, LEV 292). Most patients had Fine Pneumonia Severity Index II to IV (80.7% TGC, 74.4% LEV, mITT). At TOC (CE), TGC cured 253/282 patients (89.7%) and LEV cured 252/292 patients (86.3%); the absolute difference of TGC-LEV was 3.4% (95% confidence interval [CI], -2.2 to 9.1, noninferior [P < 0.001]). In c-mITT, TGC cured 319/394 patients (81.0%) and LEV cured 321/403 patients (79.7%); the absolute difference of TGC-LEV was 1.3% (95% CI -4.5 to 7.1, noninferior [P < 0.001]). The drug related adverse events (AEs) of nausea (20.8% TGC versus 6.6% LEV) and vomiting (13.2% TGC versus 3.3% LEV) were significantly higher in TGC; elevated alanine aminotransferase (2.8% TGC versus 7.3% LEV) and aspartate aminotransferase (2.6% TGC versus 6.9% LEV) were significantly higher in LEV. Discontinuations for AEs were low (TGC, 26 patients [6.1%]; LEV, 34 patients [8.1%]). TGC appeared safe and achieved cure rates similar to LEV in hospitalized patients with CAP. PMID- 18508228 TI - Cellular genesis in the postnatal piglet. AB - Because of the anatomical and developmental similarity of the piglet brain to the human brain we were interested in characterizing the areas of cellular genesis which occur postnatally to validate the model for subsequent neurobiological research. In this study, four piglets were injected with 5-bromodeoxyuridine (BrdU) at 6, 7 and 8 days of age. The animals were sacrificed at 13 days of age and the brains were analyzed to characterize areas of cellular genesis. BrdU was seen throughout the brain and found to be most abundant in the subventricular zone (SVZ); doublecortin (DCX) expressing cells were found throughout the white matter-with an extensive DCX network in the SVZ. Here we describe for the first time the use of immunohistochemistry for BrdU and DCX to study cellular genesis in the piglet brain. PMID- 18508229 TI - [Risk factors for cancers of the oral cavity, pharynx (cavity excluded) and larynx]. AB - OBJECTIVE: To review the risk factors for squamous cell carcinoma of the oral cavity, pharynx, and larynx. METHODS: Review of the literature using the Medline digital database (1980-2007). Previously published studies or studies not found in the database were included if relevant. Four types of studies were selected: (1) epidemiological, (2) toxicologic, (3) clinical, and (4) fundamental research. Publications concerning cancer of the nasopharynx were excluded. This work is based upon the ANAES guide for analysis of the literature and rating of guidelines, published in January 2000. RESULTS: The principal risk factors are tobacco and alcohol. Other risk factors, particularly infectious (viral) or environmental (nutritional and occupational), are also involved. From this analysis we conclude that: (1) most clinical and fundamental publications concern smoking and alcohol use; (2) studies of other risk factors are relatively old, especially those concerning nutritional and occupational factors; (3) most publications have a low level of scientific proof (grade C, levels 3 and 4). These 3 points explain the delay in the analysis of risk factors for upper aerodigestive tract (UADT) cancers. CONCLUSIONS: We must make up for this delay by prospective studies that include very large samples and use thorough and multivariate statistical analyses to estimate the impact of various toxic substances on the incidence of UADT cancer. This demands: (1) awareness on the part of all physicians who manage this type of cancer of the need to ask questions about exposure to risk factors besides than tobacco and alcohol; (2) collaboration between these physicians as well as with general practitioners, epidemiologists, nutritionists, and occupational physicians. PMID- 18508230 TI - [Mortality and prognostic factors in 146 patients with systemic lupus erythematosus in southern Tunisia]. AB - OBJECTIVES: The objective of this study was to analyze the survival rate, causes of death, and prognostic factors in patients with systemic lupus erythematosus (SLE), mainly from southern Tunisia. METHODS: This retrospective study examined the records of 146 Tunisian patients with SLE, all meeting at least 4 of the American College of Rheumatology criteria for this diagnosis. RESULTS: The mean age at presentation of these patients (126 women and 20 men) was 29.2 years (range: 6-55), and the mean duration of follow up 62 months (range: 3 days-374 months). Eleven patients (7.5%) died during follow-up. The 5- and 10-year survival rates in this cohort were both 92.1%. The most frequent causes of death were infections and disease activity, each accounting for 4 deaths. Logistic regression showed that nephrotic syndrome was an independent prognostic factor for death (p=0.032, OR=5.64). CONCLUSIONS: Our results confirmed the improvement in recent years of survival rates among SLE patients. Infections and disease activity remain the most common causes of death. Nephrotic syndrome is independent risk factor for mortality. PMID- 18508231 TI - Introduction of a self-report version of the Prescription Drug Use Questionnaire and relationship to medication agreement noncompliance. AB - The Prescription Drug Use Questionnaire (PDUQ) is one of several published tools developed to help clinicians better identify the presence of opioid abuse or dependence in patients with chronic pain. This paper introduces a patient version of the PDUQ (PDUQp), a 31-item questionnaire derived from the items of the original tool designed for self-administration, and describes evidence for its validity and reliability in a sample of patients with chronic nonmalignant pain and on opioid therapy. Further, this study examines instances of discontinuation from opioid medication treatment related to violation of the medication agreement in this population, and the relationship of these with problematic opioid misuse behaviors, PDUQ and PDUQp scores. A sample of 135 consecutive patients with chronic nonmalignant pain was recruited from a multidisciplinary Veterans Affairs chronic pain clinic, and prospectively followed over one year of opioid therapy. Using the PDUQ as a criterion measure, moderate to good concurrent and predictive validity data for the PDUQp are presented, as well as item-by-item comparison of the two formats. Reliability data indicate moderate test stability over time. Of those patients whose opioid treatment was discontinued due to medication agreement violation-related discontinuation (MAVRD) (n=38 or 28% of sample), 40% of these (n=11) were due to specific problematic opioid misuse behaviors. Based upon specificity and sensitivity analyses, a suggested cutoff PDUQp score for predicting MAVRD is provided. This study supports the PDUQp as a useful tool for assessing and predicting problematic opioid medication use in a chronic pain patient sample. PMID- 18508232 TI - Comparative cognitive profiles of obsessive-compulsive disorder and schizophrenia. AB - A body of neuropsychological research revealed cognitive impairments in patients suffering from obsessive-compulsive disorder (OCD). Only few investigations addressed the question of how specific these impairments are. The present study compared the performances of 19 subjects with OCD to 19 subjects with schizophrenia and 19 healthy controls on neuropsychological tasks across the main cognitive domains (memory, attention, visual spatial and executive functioning). For purposes of data-reduction, single test measures of the test battery applied were aggregated into eight cognitive domain scores. Contrary to our expectation we found comparable performance profiles of obsessive-compulsive (OC) and schizophrenia subjects across domains with impairments primarily affecting simple attentional skills and memory skills. However, deficits of subjects with schizophrenia were greater in magnitude than those of subjects with OCD on all domains assessed. Elevated depression scores exerted a relevant impact on performance deficits in the OC but not in the schizophrenia sample. PMID- 18508234 TI - Movement disorder and neuromuscular change in zebrafish embryos after exposure to caffeine. AB - Though caffeine is broadly distributed in many plants and foods, little is known about the teratogenic effects of caffeine during early embryonic development. Here, we used zebrafish as a model to test toxicity and teratogenicity since they have transparent eggs, making the organogenesis of zebrafish embryos easier to observe. When the exposure doses of caffeine were less than 150 ppm (17.5, 35, 50, 100 and 150 ppm), the zebrafish embryos exhibited no significant differences in survival rates after comparison with vehicle-control (0 ppm) group. As the exposure dosages increased, the survival rates decreased. No embryos survived after treatment with 300 ppm caffeine or higher dosages. The most evident change in embryos treated with caffeine was a shorter body length (vehicle-control: 3.26+/-0.01 mm, n=49; vs 150 ppm of caffeine: 2.67+/-0.03 mm, n=50). In addition, caffeine-treated embryos exhibited significantly reduced tactile sensitivity frequencies of touch-induced movement (vehicle-control: 9.93+/-0.77 vs 17.5-150 ppm caffeine: 5.37+/-0.52-0.10+/-0.06). Subtle changes are easily observed by staining with specific monoclonal antibodies F59, Znp1 and Zn5 to detect morphological changes in muscle fibers, primary motor axons and secondary motor axon projections, respectively. Our data show that the treatment of caffeine leads to misalignment of muscle fibers and motor neuron defects, especially secondary motor neuron axonal growth defects. PMID- 18508233 TI - Cocaine increases stimulated dopamine release more in periadolescent than adult rats. AB - The neural mechanisms responsible for the enhanced adolescent vulnerability for initiating drug abuse are unclear. We investigated whether age differences in dopamine neurotransmission could explain cocaine's enhanced psychomotor effects in the periadolescent rat. Electrical stimulation of the medial forebrain bundle of anesthetized post-natal age 28 days (PN28) and PN65 rats elicited dopamine release in caudate nucleus and nucleus accumbens core before and after 15 mg/kg cocaine i.p. Extracellular dopamine concentrations were greater in PN65 than PN28 caudate following 20 and 60 Hz stimulations and in the PN65 nucleus accumbens following 60 Hz stimulations. Cocaine increased dopamine concentrations elicited by 20 Hz stimulations 3-fold in the adult, but almost 9-fold in periadolescent caudate. Dopamine release rate was lower in the periadolescent caudate although total dopamine clearance was similar to that of adults. The periadolescent caudate achieved adult levels of clearance by compensating for a lower V(max) with higher uptake affinity. Tighter regulation of extracellular dopamine by the higher uptake/release ratio in periadolescents led to greater increases after cocaine. In nucleus accumbens, dopamine release and V(max) were lower in periadolescents than adults, but uptake affinity and cocaine effects were similar. Immaturity of dopamine neurotransmission in dorsal striatum may underlie enhanced acute responses to psychostimulants in adolescent rats and suggests a mechanism for the greater vulnerability of adolescent humans to drug addiction. PMID- 18508236 TI - Nerve growth factor-induced stimulation of p38 mitogen-activated protein kinase in PC12 cells is partially mediated via G(i/o) proteins. AB - Differentiation of PC12 cells by nerve growth factor (NGF) requires the activation of various mitogen-activated protein kinases (MAPKs) including p38 MAPK. Accumulating evidence has suggested cross-talk regulation of NGF-induced responses by G protein-coupled receptors, thus we examined whether NGF utilizes G(i/o) proteins to regulate p38 MAPK in PC12 cells. Induction of p38 MAPK phosphorylation by NGF occurred in a time- and dose-dependent manner and was partially inhibited by pertussis toxin (PTX). NGF-dependent p38 MAPK phosphorylation became insensitive to PTX treatment upon transient expressions of Galpha(z) or the PTX-resistant mutants of Galpha(i2) and Galpha(oA). Moreover, Galpha(i2) was co-immunoprecipitated with the TrkA receptor from PC12 cell lysates. To discern the participation of various signaling intermediates, PC12 cells were treated with a panel of specific inhibitors prior to the NGF challenge. NGF-induced p38 MAPK phosphorylation was abolished by inhibitors of Src (PP1, PP2, and SU6656) and MEK1/2 (U0126). Inhibition of the p38 MAPK pathway also suppressed NGF-induced PC12 cell differentiation. In contrast, inhibitors of JAK2, phospholipase C, protein kinase C and Ca(2+)/calmodulin-dependent kinase II did not affect the ability of NGF to activate p38 MAPK. Collectively, these studies indicate that NGF-dependent p38 MAPK activity may be mediated via G(i2) protein, Src, and the MEK/ERK cascade. PMID- 18508237 TI - 125I seed implant brachytherapy-assisted surgery with preservation of the facial nerve for treatment of malignant parotid gland tumors. AB - The surgical treatment of malignant parotid gland tumors combined with (125)I seed implant brachytherapy and preservation of the facial nerve is described. Tumor and parotid gland resection with preservation of the facial nerve was carried out in 12 patients with malignant parotid gland tumors. (125)I seeds were implanted into the target area intra- or postoperatively. The extent of regional control of the tumor was followed up, and facial nerve function was evaluated. None of the patients had tumor recurrence during the follow-up period of 50-74 months (median follow-up period, 66 months). Facial nerve function had recovered to normal by 6 months postoperatively in all patients. A limited surgical resection combined with (125)I seed implant brachytherapy is therefore considered to be an alternative treatment for local control of malignant parotid gland tumors with preservation of the facial nerve. PMID- 18508238 TI - Familial epilepsy and developmental dysphasia: description of an Italian pedigree with autosomal dominant inheritance and screening of candidate loci. AB - PURPOSE: To describe a familial epileptic condition combining a peculiar electro clinical pattern with developmental language dysfunction in a large Italian kindred. METHODS: We studied the clinical and neurophysiological features of a 4 generation family with 10 affected members (3 deceased). We also analysed in 7 affected and 7 healthy members microsatellite markers for 51 candidate loci for epilepsy, including 42 loci containing ion channel genes expressed in the brain, as well as the SPCH1 and SRPX2 loci. RESULTS: Five of the seven living affected members (aged 20-58 years) had the full phenotype (seizures, EEG epileptiform abnormalities and dysphasia). The language dysfunction was the first symptom, becoming evident since the period of language development and mainly consisting of phonemic and syntactic paraphasias, difficulty of expression and reduced verbal fluency. The seizures had their onset between 2 and 23 years and were reported as epileptic falls (4) associated or not with myoclonic features, absences (3), tonic-clonic (1) and complex partial seizures (1). The seizures were easily controlled by antiepileptic treatment in all patients except one. In the five patients with a good response of seizures to treatment, the EEG tracings showed the coexistence of focal and generalized epileptiform abnormalities; in the refractory patient the interictal EEG demonstrated bilateral asynchronous fronto-temporal paroxysms with left predominance and ictal SEEG recording suggested a multifocal origin of the discharges. MRI of the brain was normal in all patients. Linkage analysis provided negative LOD scores for all the investigated loci. CONCLUSION: We have described a novel familial pattern of epilepsy and developmental dysphasia which is not genetically linked to epilepsy or speech disorder loci, as documented by a candidate-gene linkage approach. PMID- 18508239 TI - The effect of epileptic seizures on proton MRS visible neurochemical concentrations. AB - PURPOSE: To investigate post-ictal changes in cerebral metabolites. METHODS: We performed a longitudinal quantitative proton magnetic resonance spectroscopy (MRS) study in 10 patients with epilepsy and 10 control subjects. The patients were studied on two occasions: immediately following a seizure, and on a second occasion at least 7h after the most recent seizure. Each study measured N-acetyl aspartate plus N-acetyl aspartyl glutamate (NAAt), Creatine plus phosphocreatine (Cr), Choline containing compounds (Cho) and glutamate plus glutamine (GLX) concentrations using a short-echo time sequence (TE=30ms), and NAAt, Cr and lactate using a second sequence with longer echo time (TE=144ms). The control group was studied on two occasions using the same sequences. RESULTS: No inter scan differences were observed for the control group. NAAt and NAAt/Cr levels were lower in the patient group at both measured TEs but did not change significantly between studies. The ratio of Cr at TE 144ms to TE 30ms (Cr(144)/Cr(30)) and GLX/Cr were higher and Cho lower in the post-ictal scan compared to the inter-ictal study. Change in Cr(144)/Cr(30) and NAAt(144)/Cr(144) correlated with the post-ictal interval. Lactate measurement at longer TE was not informative. DISCUSSION: Proton MRS is sensitive to metabolite changes following epileptic seizures within the immediate post-ictal period. The ratio Cr(144)/Cr(30) is the most sensitive measure of metabolic disturbance and is highest in the post-ictal period but appears to normalise within 2h of the most recent seizure. PMID- 18508240 TI - Glial cell number and nuclear size in the mediodorsal thalamic nucleus (MDNT) in schizophrenia. PMID- 18508242 TI - Association between interleukin-6 receptor polymorphism and patients with schizophrenia. PMID- 18508241 TI - Analysis of protocadherin alpha gene enhancer polymorphism in bipolar disorder and schizophrenia. AB - Cadherins and protocadherins are cell adhesion proteins that play an important role in neuronal migration, differentiation and synaptogenesis, properties that make them targets to consider in schizophrenia (SZ) and bipolar disorder (BD) pathogenesis. Consequently, allelic variation occurring in protocadherin and cadherin encoding genes that map to regions of the genome targeted in SZ and BD linkage studies are particularly strong candidates to consider. One such set of candidate genes is the 5q31-linked PCDH family, which consists of more than 50 exons encoding three related, though distinct family members--alpha, beta, and gamma--which can generate thousands of different protocadherin proteins through alternative promoter usage and cis-alternative splicing. In this study, we focused on a SNP, rs31745, which is located in a putative PCDHalpha enhancer mapped by ChIP-chip using antibodies to covalently modified histone H3. A striking increase in homozygotes for the minor allele at this locus was detected in patients with BD. Molecular analysis revealed that the SNP causes allele specific changes in binding to a brain protein. The findings suggest that the 5q31-linked PCDH locus should be more thoroughly considered as a disease susceptibility locus in psychiatric disorders. PMID- 18508243 TI - An MRI and proton spectroscopy study of the thalamus in children with autism. AB - Thalamic alterations have been reported in autism, but the relationships between these abnormalities and clinical symptoms, specifically sensory features, have not been elucidated. The goal of this investigation is to combine two neuroimaging methods to examine further the pathophysiology of thalamic anomalies in autism and to identify any association with sensory deficits. Structural MRI and multi-voxel, short echo-time proton magnetic resonance spectroscopy ((1)H MRS) measurements were collected from 18 male children with autism and 16 healthy children. Anatomical measurements of thalamic nuclei and absolute concentration levels of key (1)H MRS metabolites were obtained. Sensory abnormalities were assessed using a sensory profile questionnaire. Lower levels of N-acetylaspartate (NAA), phosphocreatine and creatine, and choline-containing metabolites were observed on the left side in the autism group compared with controls. No differences in thalamic volumes were observed between the two groups. Relationships, although limited, were observed between measures of sensory abnormalities and (1)H MRS metabolites. Findings from this study support the role of the thalamus in the pathophysiology of autism and more specifically in the sensory abnormalities observed in this disorder. Further investigations of this structure are warranted, since it plays an important role in information processing as part of the cortico-thalamo-cortical pathways. PMID- 18508245 TI - Automated gray level index measurements reveal only minor cytoarchitectonic changes of Brodmann area 9 in schizophrenia. AB - Using an automatized gray level index (GLI) method, we recently found cytoarchitectonic abnormalities in schizophrenia in Brodmann area 10 (BA10) [Vogeley, K., Tepest, R., Schneider-Axmann, T., Hutte, H., Zilles, K., Honer, W.G., Falkai, P., 2003. Automated image analysis of disturbed cytoarchitecture in Brodmann area 10 in schizophrenia, Schizophrenia Research 62, 133-140]. As another potential key region involved in the pathophysiology of schizophrenia, we have now investigated BA9 in the same sample consisting of 20 schizophrenic cases and 20 controls. The GLI value represents the area-percentage covered by perikarya in measuring fields of microscopic images. BA9 was analyzed with respect to the factors diagnosis and gender for six different compartments approximately corresponding to the neocortical layers. The main result in BA9 was a significant interaction of diagnosis and gender for GLI in layers IV and V on the left side. Subsequent analyses separately performed concerning gender revealed a significant GLI increase in layer V on the left side in male patients compared with controls. However, after an adjustment of error probabilities for multiple testing, differences did not reach significance. No GLI difference was observed in the sample between diagnostic groups for females and between the diagnostic groups in general. Comparisons with our BA10 results suggest that cytoarchitectural changes relevant to schizophrenia appear different in various Brodmann areas. Since increases in GLI were found only in selected layers (V and VI) of BA9, these findings do not support a generalized neuropil reduction across all cortical layers. PMID- 18508244 TI - Hippocampal volumes and depression subtypes. AB - Studies of depression and hippocampal volume have yielded inconsistent results. This inconsistency could stem from the heterogeneity of depressive disorders. We conducted cross-sectional and longitudinal analyses of hippocampal volumes in atypical depressive, melancholic depressive, and control subjects. We found no effect of depression subtype on hippocampal volume or memory performance. PMID- 18508246 TI - Voltammetric detection of magnolol in Chinese medicine based on the enhancement effect of mesoporous Al/SiO(2)-modified electrode. AB - The enhancement effect of mesoporous Al-doped silica (Al/SiO(2))-modified electrode was investigated. Due to the properties such as large surface area, strong adsorptive ability and numerous active sites, mesoporous Al/SiO(2) alters the structure and property of electrode/solution interface, then greatly improves the electrochemical response of magnolol. The electrochemical behavior of magnolol was examined in detail. It is found that the oxidation peak current of magnolol remarkably increases at the mesoporous Al/SiO(2)-modified electrode. Based on this, a sensitive and convenient electrochemical method was developed for the determination of magnolol. The linear range is over the range from 7.5 x 10(-8) to 2.0 x 10(-5) mol L(-1). The limit of detection (S/N=3) is as low as 2.5 x 10(-8) mol L(-1). Finally, this novel method was successfully used to determine the magnolol in Chinese traditional medicines. PMID- 18508247 TI - Antibacterial mechanism of chitosan microspheres in a solid dispersing system against E. coli. AB - In this study, we investigated the antibacterial mechanism through the interfacial contacting inhibition behaviors of chitosan antimicrobials against Escherichia coli in solid dispersing state. Chitosan microspheres (CMs) were prepared by emulsification cross-linking reaction, and oleoyl-CMs (OCMs) were obtained by introduction of oleoyl groups to the chitosan. The CMs were with smooth surface and spherical shape of diameter of about 124 microm. The antibacterial activity was directly proportional to the concentration and the hydrophobic property of CMs. The fluorescence experiments indicated CMs had influenced the structure of membrane, especially the OCMs were speculated to interact with proteins on the cell membrane. SEM photographs showed E. coli adhered to the surface of the CMs and provided evidences for the disruption of the cells, while the bacterium conglomerated on the surface of the OCMs. The CMs changed the permeability of membrane and caused cellular leakage that correlated with the hydrophobic interaction between CMs and cytoplasmic membrane phospholipids of Gram-negative bacteria. Solid dispersing system makes the antibacterial activities of CMs counted as a sequent event-driven to study the antibacterial mechanism of chitosan originally. PMID- 18508248 TI - [Moyamoya and Down syndrome: about 2 cases]. AB - Moyamoya syndrome has rarely been reported in association with Down syndrome. We report on 2 cases in 3-year-old and 6-year-old female children with Down syndrome, who presented with neurological deficit. Imaging (magnetic-resonance angiography and digital-subtraction angiography) revealed the classical Moyamoya pattern. The neurological deficits persisted in both cases. One patient has developed epilepsy. PMID- 18508249 TI - [Hypertension in the absence of renal involvement during childhood Henoch Schonlein purpura]. AB - Henoch-Schonlein purpura (HSP) is an IgA mediated vasculitis that affects small vessels. In this condition, arterial hypertension is most often linked with renal involvement, but it can also occur in the absence of urinary abnormalities. We report the case of a 12-year-old girl who presented with HSP and hypertension in the absence of renal involvement. Hypertension responded to oral therapy with nicardipine, and disappeared after healing of the disease. In the case of hypertension during HSP in the absence of renal involvement, other causes of childhood's arterial hypertension have to be ruled out. PMID- 18508250 TI - [Answer to the article: "comparison of prescriptions by pediatricians and general practitioners: a population-based study in Franche-Comte from the database of Regional Health Insurance Fund" Arch Pediatr 2005;12:1688-9]. PMID- 18508251 TI - Regulatory T cells and treatment of cancer. AB - CD4+CD25+FOXP3+ regulatory T cells (Tregs) are elevated in cancers and can thwart protective antitumor immunity. Recent human cancer trials suggest that depleting Tregs can be clinically beneficial. Additional types of deleterious regulatory cells are also increased in cancer. Tregs also play unanticipated roles in cancer therapy in that some drugs unexpectedly increase (e.g. cancer vaccines or IL-2 treatment) or decrease (e.g. antineoangiogenesis agents or receptor tyrosine kinase inhibitors) their numbers or function. Managing deleterious effects of regulatory cells represents a novel and potentially effective way to give immunotherapy for cancer. New insights into molecular mechanisms governing trafficking, differentiation, and function of these cells suggest novel approaches to manipulating them as treatment strategies. PMID- 18508252 TI - New twists in X-chromosome inactivation. AB - Dosage compensation, the mechanism by which organisms equalize the relative gene expression of dimorphic sex chromosomes, requires action of a diverse range of epigenetic mechanisms. The mammalian form, 'named X-chromosome inactivation' (XCI), involves silencing of one X chromosome in the female cell and regulation by genes that make noncoding RNAs (ncRNA). With large-scale genomic and transcriptome studies pointing to a crucial role for noncoding elements in organizing the epigenome, XCI emerges as a major paradigm and a focus of active research worldwide. With more surprising twists, recent advances point to the significance of RNA-directed chromatin change, chromosomal trans-interactions, nuclear organization, and evolutionary change. These findings have impacted our understanding of general gene regulation and are discussed herein. PMID- 18508253 TI - Molecular implementation and physiological roles for histone H3 lysine 4 (H3K4) methylation. AB - Chromosomal surfaces are ornamented with a variety of post-translational modifications of histones, which are required for the regulation of many of the DNA-templated processes. Such histone modifications include acetylation, sumoylation, phosphorylation, ubiquitination, and methylation. Histone modifications can either function by disrupting chromosomal contacts or by regulating non-histone protein interactions with chromatin. In this review, recent findings will be discussed regarding the regulation of the implementation and physiological significance for one such histone modification, histone H3 lysine 4 (H3K4) methylation by the yeast COMPASS and mammalian COMPASS-like complexes. PMID- 18508255 TI - Phytoglycoprotein (24 kDa) inhibits expression of PCNA via PKCalpha and MAPKs in oxygen radical-stimulated Chang liver cells. AB - The purpose of this study was to investigate the inhibitory effect of 24-kDa glycoprotein isolated from Zanthoxylum piperitum DC fruit (ZPDC glycoprotein) on glucose/glucose oxidase (G/GO)- or hypoxanthine/xanthine oxidase (HX/XO)-induced cell proliferation in Chang liver cells. We found that ZPDC glycoprotein has significant scavenging effect on the production of intracellular H2O2 without cytotoxicity in G/GO- or HX/XO-treated in Chang liver cells. In the G/GO or HX/XO stimulated protein kinases activity, ZPDC glycoprotein inhibited translocation of protein kinase C alpha (PKCalpha) to membrane and phosphorylation of extracellular signal-regulated kinase, p38 MAP kinase and c-Jun N-terminal kinase, respectively. In the G/GO or HX/XO-stimulated transcriptional activity, ZPDC glycoprotein also blocked the DNA binding activities of nuclear factor-kappa B and activator protein-1 and attenuated the activities of p50, p65, c-Jun and c Fos, respectively. Finally, in the G/GO or HX/XO-stimulated cell proliferation, the activity of proliferating cell nuclear antigen was significantly blocked by treatment with ZPDC glycoprotein as well as protein kinase C inhibitor and mitogen-activated protein kinase inhibitors. On the basis of these results, we speculate that this glycoprotein is one of the natural antioxidants and of the modulators on abnormal activation of cell proliferation-related molecules in Chang liver cells. PMID- 18508256 TI - Fentanyl use among street drug users in Toronto, Canada: behavioural dynamics and public health implications. AB - Prescription opioids (POs) are playing an increasingly central role in street drug use and related harms in North America. One distinct PO substance of interest is Fentanyl (Duragesic), a potent opioid analgesic designed for transdermal time-release application. Studies from Europe and North America have documented the sizeable overdose and mortality burden associated with the non medical use of this drug. This study explores practices and risk dynamics associated with Fentanyl abuse, also considering public health implications. Semi structured interviews were conducted with 25 regular street-entrenched illicit PO users in Toronto, Canada, a sub-sample of which were recent Fentanyl users. Results showed that while relatively rare on the illicit PO market in Toronto, Fentanyl is a highly desired, sought after and relatively expensive PO drug among street users. In addition, the new 'matrix' patch technology implemented for Fentanyl since 2005 is a limited safeguard against abuse as simple extraction methods are utilized by street users. Finally, distinct risk behaviours relevant for public health emerge due to the high black market costs of Fentanyl and the extraction techniques applied, potentially facilitating high risks for infectious disease (e.g., HCV, HIV) transmission and/or overdose. Consequently, prevalence and practices of Fentanyl use by street users require closer monitoring, targeted interventions and further research regarding risks and outcomes. PMID- 18508257 TI - New tools for the study and direct surveillance of viral pathogens in water. AB - Half a century ago scientists attempted the detection of poliovirus in water. Since then other enteric viruses responsible for gastroenteritis and hepatitis have replaced enteroviruses as the main target for detection. However, most viral outbreaks are restricted to norovirus and hepatitis A virus, making them the main targets in water. The inclusion of virus analysis in regulatory standards for viruses in water samples must overcome several shortcomings such as the technical difficulties and high costs of virus monitoring, the lack of harmonised and standardised assays and the challenge posed by the ever-changing nature of viruses. However, new tools are nowadays available for the study and direct surveillance of viral pathogens in water that may contribute to fulfil these requirements. PMID- 18508258 TI - Ena/VASP: proteins at the tip of the nervous system. AB - The emergence of neurites from a symmetrical cell body is an essential feature of nervous system development. Neurites are the precursors of axons and dendrites and are tipped by growth cones, motile structures that guide elongating axons in the developing nervous system. Growth cones steer the axon along a defined path to its appropriate target in response to guidance cues. This navigation involves the dynamic extension and withdrawal of actin-filled finger-like protrusions called filopodia that continuously sample their environment. Ena/VASP proteins, a conserved family of actin-regulatory proteins, are crucial for filopodia formation and function downstream of several guidance cues. Here we review recent findings into Ena/VASP function in neurite initiation, axon outgrowth and guidance. PMID- 18508259 TI - Function and regulation of local axonal translation. AB - An increasing body of evidence indicates that local axonal translation is required for growing axons to respond appropriately to guidance cues and other stimuli. Recent studies suggest that asymmetrical synthesis of cytoskeletal proteins mediates growth cone turning and that local translation and retrograde transport of transcription factors mediate neuronal survival. Axonal translation is regulated partly by selective axonal localization of mRNAs and by translation initiation factors and RNA-binding proteins. We discuss possible rationales for local axonal translation, including distinct properties of nascent proteins, precise localization, and axonal autonomy. PMID- 18508260 TI - The determination of projection neuron identity in the developing cerebral cortex. AB - Here we review the mechanisms that determine projection neuron identity during cortical development. Pyramidal neurons in the mammalian cerebral cortex can be classified into two major classes: corticocortical projection neurons, which are concentrated in the upper layers of the cortex, and subcortical projection neurons, which are found in the deep layers. Early progenitor cells in the ventricular zone produce deep layer neurons that express transcription factors including Sox5, Fezf2, and Ctip2, which play important roles in the specification of subcortically projecting axons. Upper layer neurons are produced from progenitors in the subventricular zone, and the expression of Satb2 in these differentiating neurons is required for the formation of axonal projections that connect the two cerebral hemispheres. The Fezf2/Ctip2 and Satb2 pathways appear to be mutually repressive, thus ensuring that individual neurons adopt either a subcortical or callosal projection neuron identity at early times during development. The molecular mechanisms by which Satb2 regulates gene expression involves long-term epigenetic changes in chromatin configuration, which may enable cell fate decisions to be maintained during development. PMID- 18508261 TI - Agonist-like SERM effects on ERalpha-mediated repression of MMP1 promoter activity predict in vivo effects on bone and uterus. AB - Estradiol receptors (ER), ERalpha and ERbeta, are ligand-dependent transcription factors that regulate gene expression. Human and murine genetics suggest that ERalpha is the key target for estradiol action on bone, uterus and breast. To date, the molecular mode of action of estradiol and selective estradiol receptor modulators (SERMs) on bone is not fully understood. This is exemplified by a lack of in vitro assays that reliably predict SERM agonist activities in vivo. We hypothesized that ligand-dependent ERalpha transrepression, via protein-protein interactions at AP1, may predict estrogenic effects on bone. We modeled this using the MMP1 promoter, which encodes an AP1 binding site. We show that ICI 182780, raloxifene, 4-hydroxytamoxifen and estradiol all exhibit differential agonistic activities on the MMP1 promoter by suppressing activity by 20-80%. Transrepression efficacy and potency correlated with both uterotrophic (R(2)=0.98) and osteoprotective (R(2)=0.80) potential in the ovariectomized rat. This identifies MMP1 promoter transrepression as an agonist activity commonly shared by AF2 agonists and "antagonists" alike. Mutation analysis showed that the repression by estradiol and SERMs required correct amino acid sequences in the AF 2 domain. For instance, L540Q AF2 mutation did not alter responses to raloxifene, although it greatly increased responses to ICI-182780 (threefold) and reduced estradiol's effect by 20%. Furthermore, all tested ligands repressed the MMP1 promoter through the L540Q mutant with identical efficacy. Together, these data suggest that estradiol and SERMs share common agonist transcriptional activity via protein-protein interactions at AP1. PMID- 18508262 TI - Soil contamination by heavy metals: measurements from a closed unlined landfill. AB - The aim of the present study was the characterization of soil samples of a closed unlined landfill located northwest of Thessaloniki, North Greece, in relation to heavy metals values. Samples were obtained by drilling in different depths (2.5 17.5m). Then they were analyzed by atomic absorption spectrophotometry for Cd, Cr, Cu, Ni, Pb and Zn investigation. The chemical analysis showed that the metal values varied over a wide range: from 0.50 to 18.75mg/kg for Cd, 3.88-171.88mg/kg for Cr, 8.13-356.25mg/kg for Cu, 5.63-63.75mg/kg for Ni, 2.50-92.50mg/kg for Pb and 6.38-343.75mg/kg for Zn. The highest values found in three of the six drillings, in depths over 2.5m. Although the area is heavily industrialized, the presented results indicated that local industries have not constituted an extensive metal pollution source for the site. Finally, after all necessary preparatory operations of site cleaning and flattening, surface planting selected and applied as a phytoremediation rehabilitation method of the site. PMID- 18508263 TI - Pretreatment of woody and herbaceous biomass for enzymatic saccharification using sulfuric acid-free ethanol cooking. AB - A sulfuric acid-free ethanol cooking (SFEC) treatment was developed to achieve complete saccharification of the cellulosic component of eucalyptus and baggase flour, thereby avoiding the problems associated with the use of strong acid catalysts. Cutter-milled flours were exposed to an ethanol (EtOH)/water/acetic acid mixture in an autoclave. Enzymatic hydrolysis experiments of the pretreated samples demonstrated that almost complete conversion of the cellulosic components to glucose was achieved under optimal conditions. A large-scale trial revealed that there was little consumption of in-feed EtOH during SFEC; therefore, it is considered that most part EtOH used can be essentially recovered and reused. Field emission scanning electron microscopy showed that SFEC induced the formation of pores ranging in size from approximately 10 to several 100nm. It can be assumed that the porous surface was due to the partial removals of lignin and hemicellulose, which improved the accessibility of the enzyme onto the substrate. PMID- 18508264 TI - Discovery of novel 4-amino-6-arylaminopyrimidine-5-carbaldehyde oximes as dual inhibitors of EGFR and ErbB-2 protein tyrosine kinases. AB - We herein disclose a novel series of 4-aminopyrimidine-5-carbaldehyde oximes that are potent and selective inhibitors of both EGFR and ErbB-2 tyrosine kinases, with IC(50) values in the nanomolar range. Structure-activity relationship (SAR) studies elucidated a critical role for the 4-amino and C-6 arylamino moieties. The X-ray co-crystal structure of EGFR with 37 was determined and validated our design rationale. PMID- 18508265 TI - TrkA kinase inhibitors from a library of modified and isosteric Staurosporine aglycone. AB - An immobilized Staurosporine aglycone isostere where one of the indole nitrogen atoms was replaced by carbon has been sequentially functionalized to generate compounds inhibiting TrkA kinase. In the first phase, initial screening of a library of C13-hydroxymethyl-7-oxo-indenopyrrolocarbazoles resulted in several potent compounds, one of which was further optimized to generate the corresponding carbamates on solid phase. Some of the major carbamate diastereomers were found to be several-fold more potent than their alcohol parents. Synthesis, SAR analysis, kinase selectivity, and anti-tumor properties of a TrkA inhibitor (12a) are discussed. PMID- 18508266 TI - Novel mutations in the TK2 gene associated with fatal mitochondrial DNA depletion myopathy. AB - Mitochondrial DNA depletion syndromes are a heterogeneous group of childhood neurological disorders characterised by a quantitative abnormality of mitochondrial DNA. We describe two siblings who presented at 8 months and 14 months with myopathy, which rapidly progressed and resulted in death by respiratory failure at age 14 and 18 months, respectively. Muscle biopsy revealed marked respiratory chain defects, with real-time PCR confirming a dramatic depletion of mitochondrial DNA. Sequencing of the thymidine kinase 2 (TK2) gene revealed two, novel heterozygous mutations (p.Q87X and p.N100S) with parental DNA analysis confirming the transmission of mutated alleles. PMID- 18508267 TI - Eight years experience with enzyme replacement therapy in two children and one adult with Pompe disease. AB - Pompe disease (type 2 glycogenosis, acid maltase deficiency) is a disorder affecting skeletal and cardiac muscle, caused by deficiency of acid alpha glucosidase. In 2006 enzyme therapy with recombinant human alpha-glucosidase received marketing approval based on studies in infants. Results in older children and adults are awaited. Earlier we reported on the 3-year follow-up data of enzyme therapy in two adolescents and one adult. In the present study these patients were followed for another 5 years. Two severely affected patients, wheelchair and ventilator dependent, who had shown stabilization of pulmonary and muscle function in the first 3 years, maintained this stabilization over the 5 year extension period. In addition patients became more independent in daily life activities and quality of life improved. The third moderately affected patient had shown a remarkable improvement in muscle strength and regained the ability to walk over the first period. He showed further improvement of strength and reached normal values for age during the extension phase. The results indicate that both long-term follow-up and timing of treatment are important topics for future studies. PMID- 18508268 TI - Vascular endothelial growth factor is a strong predictor of early distant recurrences in a prospective study of premenopausal women with lymph-node negative breast cancer. AB - We investigate the prognostic significance of the pro-angiogenic cytokine vascular endothelial growth factor (VEGF), urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor 1 (PAI-1), and S-phase fraction (SPF) for distant disease free survival (DDFS) in 219 premenopausal patients with node negative breast cancer (NNBC). In univariate analysis significantly shorter DDFS was observed for patients with high VEGF (p=0.006), high uPA (p=0.001), and high SPF (p<0.001). The prognostic significance of VEGF varied over time being very strong for early relapses (0-2.25 years follow-up) (HR=7.9; p=0.006) while no difference was seen in the subsequent follow-up period (HR=1.3; p=0.62). In a series of bivariate analyses VEGF provided prognostic information during the whole observation period (0-72 months) in addition to age, tumour size, oestrogen receptor (ER), progesterone receptor (PgR), and uPA. Also this effect was more pronounced during the first follow-up period suggesting VEGF as a marker of early recurrences. PMID- 18508269 TI - Lost to translation: when autophagy targets mature ribosomes. AB - Autophagy and the ubiquitin proteasome system (UPS) mediate the degradation of cellular proteins. However, we are now realizing that autophagy can also be a selective process that degrades various organelles. Peter and co-workers discovered a selective autophagic pathway that targets ribosomes in Saccharomyces cerevisiae. This pathway, which they termed ribophagy, depends on Ubp3 ubiquitin protease and its partner Bre5. This is an important finding, because it suggests that the number of ribosomes can be adjusted to match the needs of the cell. PMID- 18508270 TI - Characterising the clinical and biomechanical features of severely deformed feet in rheumatoid arthritis. AB - PURPOSE: Foot deformity is a well-recognised impairment in patients with rheumatoid arthritis (RA) which results in functional disability. Deformity can occur at the rearfoot, midfoot, forefoot or in combination and the impact that site-specific foot deformities has on functional disability is largely unknown. The aim of this study was to describe the clinical and biomechanical characteristics of patients with severe rearfoot, forefoot or combined deformities and determine localised disease impact. METHODS: Twenty-eight RA patients with severe forefoot (FF group n=12), rearfoot (RF group n=10) or combined deformities (COMB group n=6) were recruited. Each patient underwent 3D gait analysis and plantar pressure measurements. Localised disease impact and foot-specific disease activity were determined using the Leeds Foot Impact Scale and clinical examination respectively. Comparison was made against a normative control group (n=53). RESULTS: Patients in the COMB group walked slowest and the double-support time was longer in the RF and COMB groups compared to those in the FF group. Patients in the RF and COMB group had higher levels of foot-related disability and demonstrated excessive rearfoot eversion and midfoot collapse compared to those in the FF group. Forefoot deformity was associated with reduced toe contact, high forefoot pressures and delayed heel lift. CONCLUSIONS: Abnormal gait patterns were identified and were distinguishable among those patients with predominantly forefoot, rearfoot or combined foot deformity. PMID- 18508271 TI - The role of complex carbohydrate catabolism in the pathogenesis of invasive streptococci. AB - Historically, the study of bacterial catabolism of complex carbohydrates has contributed to understanding basic bacterial physiology. Recently, however, genome-wide screens of streptococcal pathogenesis have identified genes encoding proteins involved in complex carbohydrate catabolism as participating in pathogen infectivity. Subsequent studies have focused on specific mechanisms by which carbohydrate utilization proteins might contribute to the ability of streptococci to colonize and infect the host. Moreover, transcriptome and biochemical analyses have uncovered novel regulatory pathways by which streptococci link environmental carbohydrate availability to virulence factor production. Herein we review new insights into the role of complex carbohydrates in streptococcal host-pathogen interaction. PMID- 18508272 TI - Synthetic and pharmacological studies on new simplified analogues of the potent actin-targeting Jaspamide. AB - In the recent years, we focused our attention on the cyclodepsipeptide Jaspamide 1, an interesting marine metabolite, possessing a potent inhibitory activity against breast and prostate cancer, as a consequence of its ability to disrupt actin cytoskeleton dynamics. Although its biological profile has been well determined, many mechanistic details are still missing in terms of molecular target identification. For this reason, we decided to synthetically modify the natural metabolite, obtaining small arrays of unnatural variants useful to illuminate the structural requirements essential for the activity. Here, we report the synthesis of seven new Jaspamide analogues 2-8, containing, as the parent compound, a beta-amino acid in the cyclopeptide backbone. Their biological profile is also described. PMID- 18508273 TI - Synthesis and antitumor activity evaluations of albumin-binding prodrugs of CC 1065 analog. AB - An albumin-binding prodrug of the extremely potent CC-1065 analog, (+)-FDI-CBI, has been synthesized. This analog, (+)-FDI-CBIM, formed an albumin conjugate when added to human albumin in vitro. A greater amount (>3-fold) of the prodrug can be administered to animals compared to the free drug. The prodrug had significantly improved antitumor efficacy compared to the free drug in animal models using syngeneic animal tumors and human ovarian xenografted tumor cells. Antitumor drug delivery by in situ formation of drug-albumin conjugate is a promising strategy to improve antitumor efficacy. PMID- 18508274 TI - Exposures from environmental radioactivity: international safety standards. AB - The paper presents the current international system for the protection of the public against environmental radioactivity. The protection system applies to all the three human exposure situations, i.e., planned, emergency and existing exposures. Radiation protection is a developing scientific and practical discipline and some of the areas in public radiation protection and protection of the environment that are in need of further elaboration are identified in the paper. PMID- 18508275 TI - Setting radon-specific release criteria and demonstrating compliance for land affected by NORM. AB - Residues from industrial activities involving naturally occurring radioactive materials (NORMs) may cause radiation exposures to members of the public, particularly when NORM-affected land is brought into residential use. To provide an adequate protection against radiation in such situations, the following limiting criteria are currently required in Spain for releasing NORM-affected land: (i) no more than a 300 microSv yr(-1) increase (excluding radon doses) over the natural background; (ii) (222)Rn concentrations in hypothetical future dwellings lower than 200 Bq m(-3); and (iii) reduction of all radiation exposures to as low as reasonable achievable. This paper addresses some of the problems encountered in translating the (222)Rn criterion into site-specific release limits and in demonstrating compliance with them. PMID- 18508276 TI - How to introduce radioactive chlorine into a benzene ring using [*Cl]Cl-? AB - The aim of this study was how to introduce radioactive chlorine (*Cl) isotope into a benzene ring to afford *Cl-labeled chlorobenzene ([*Cl]1). Using tributylphenylstannyl compound (2) as a starting material, we determined two routes for introducing *Cl into the benzene ring: reaction of [*Cl]Cl(-) with 2 in the presence of chloramine-T and nucleophilic reaction of [*Cl]Cl(-) with diphenyliodonium tosylate (3). The two routes provide suitable tools to prepare [*Cl]1 without electron-abstracting group in the benzene ring. PMID- 18508277 TI - Endoscopic vs open saphenous vein harvest for coronary artery bypass grafting: a prospective randomized trial. AB - OBJECTIVE: Endoscopic saphenous vein harvesting (EVH) for coronary artery bypass grafting (CABG) has been developed to reduce leg wound morbidity and improve patient satisfaction. Choosing between EVH of a short vein segment from the thigh and open venous harvesting (OVH) of a short segment from the calf represents a clinical dilemma as EVH is easiest to perform from the thigh and OVH is easiest to perform from the calf. The purpose of this study was to investigate whether leg wound morbidity was reduced after EVH of a short vein segment from the thigh compared with OVH from the calf. Secondly we investigated whether EVH would reduce length of hospital stay and improve cosmetic results. METHODS: From April 2004 to June 2007, 132 patients undergoing elective isolated CABG were randomized to have a short segment of saphenous vein harvested either by the EVH or OVH technique. Clinical follow-up was scheduled at day 5 and at 1 month. Primary end points included wound morbidity. Secondary end-points included harvest time, length of hospital stay, cosmetic results and need for additional wound care after discharge. RESULTS: The groups were preoperative similar. Three patients in the OVH group were excluded from the study as it became apparent that it was necessary to extend the incision beyond the knee. Harvest time was longer for the EVH group, but these patients suffered from significantly fewer cases of infectious and non-infective wound complications, with a substantial reduction in the need for post-discharge leg wound care. The purulent infection rates in the EVH and OVH groups were 0% and 11%, respectively. The overall leg wound morbidity rates regarding cellulitis, purulent infection, dehiscence and skin necrosis were 3% and 27% in the EVH and OVH groups, respectively (p<0.001). The length of hospital stay was similar. The conversion rate from EVH to OVH was 14%. The EVH group experienced less pain and better cosmetic results. CONCLUSIONS: EVH of a short vein segment from the thigh results in less wound morbidity and better cosmetic results compared with OVH of a short vein segment from the calf. PMID- 18508278 TI - New insights of dimethyl sulphoxide effects (DMSO) on experimental in vivo models of nociception and inflammation. AB - DMSO is one of the most common solvents used experimentally to dissolve hydrophobic substances for in vivo and in vitro purposes. A wide range of pharmacological effects exerted by DMSO has been documented in both animal and human experimental models. However, only a few and sometimes contrasting data about the effects of DMSO in animal models of nociception and inflammation are presently available. In this study, we evaluated the effects induced by DMSO and a DMSO-containing saline on thermal and chemical nociception, inflammation and locomotor activity in CD1 mice. We demonstrated that centrally or orally administered DMSO displayed anti-nociceptive effects to thermal (hot plate and tail-flick test) and chemical (formalin test) stimuli. Conversely, DMSO was able to increase both nociceptive phases in the formalin test when applied subcutaneously in the dorsal surface of the mouse hind paws 10 min before formalin administration. Oral administration of DMSO produced anti-inflammatory effects on zymosan-induced edema in the mouse paw, whereas local administration potentiated the inflammatory action exerted by zymosan. Oral and central, but not local, administration of DMSO improved the mouse locomotor activity. These results suggest that DMSO displayed opposite effects on nociception and inflammation, depending on the route of administration. New and helpful evidence about DMSO laboratory applications need to be considered in the in vivo studies to assess correctly the pharmacological properties of investigated drugs. PMID- 18508279 TI - Expression, purification, and initial characterization of human alanine aminotransferase (ALT) isoenzyme 1 and 2 in High-five insect cells. AB - Alanine aminotransferase (ALT) is a key enzyme for gluconeogenesis as well as a widely used serum marker for liver injury. We have identified two ALT isoenzymes, ALT1 and ALT2, which are encoded by separate genes. In this study, we described the expression, purification and initial characterization of human ALT1 and ALT2 proteins in High-five insect cells. Human ALT1 and ALT2 were expressed as His tagged fusion proteins by recombinant baculovirus in insect cells and purified into homogeneity in one step by using immobilized Ni2+-affinity chromatography. Tag-free ALT1 and ALT2 were obtained by cleavage of enterokinase digestion and used for initial characterization of the enzymes. The specific ALT activity of purified fusion or His-tag-removed ALT1 was about 15-fold higher than that of ALT2 and their enzymatic activities decreased quickly at 37 degrees C and -20 degrees C, but were well preserved at -80 degrees C. Nevertheless, the ALT1 and ALT2 activities remained stable in a buffer containing 25% glycerol. The pH profile was similar between hALT1 and hALT2 in that both enzymes remained fully active between pH 6.5 and 8.0. The purified ALT recombinant proteins can not only be used as a reference protein standard for the ALT assay in clinical chemistry, but also will be useful for understanding the biochemical and biological significance of the isoenzymes and for developing ALT isoform-specific assays for clinical or preclinical diagnostic use. PMID- 18508280 TI - The 32kDa enamelin undergoes conformational transitions upon calcium binding. AB - The 32 kDa hydrophilic and acidic enamelin, the most stable cleavage fragment of the enamel specific glycoprotein, is believed to play vital roles in controlling crystal nucleation or growth during enamel biomineralization. Circular dichroism and Fourier transform infrared spectra demonstrate that the secondary structure of the 32 kDa enamelin has a high content of alpha-helix (81.5%). Quantitative analysis on the circular dichroism data revealed that the 32 kDa enamelin undergoes conformational changes with a structural preference to beta-sheet with increasing concentration of calcium ions. We suggest that the increase of beta sheet conformation in the presence of Ca(2+) may allow preferable interaction of the 32 kDa enamelin with apatite crystal surfaces during enamel biomineralization. The calcium association constant (K(a)=1.55 (+/-0.13)x10(3)M( 1)) of the 32 kDa enamelin calculated from the fitting curve of ellipticity at 222 nm indicated a relatively low affinity. Our current biophysical studies on the 32 kDa enamelin structure provide novel insights towards understanding the enamelin-mineral interaction and subsequently the functions of enamelin during enamel formation. PMID- 18508281 TI - The leptospiral antigen Lp49 is a two-domain protein with putative protein binding function. AB - Pathogenic Leptospira is the etiological agent of leptospirosis, a life threatening disease that affects populations worldwide. Currently available vaccines have limited effectiveness and therapeutic interventions are complicated by the difficulty in making an early diagnosis of leptospirosis. The genome of Leptospira interrogans was recently sequenced and comparative genomic analysis contributed to the identification of surface antigens, potential candidates for development of new vaccines and serodiagnosis. Lp49 is a membrane-associated protein recognized by antibodies present in sera from early and convalescent phases of leptospirosis patients. Its crystal structure was determined by single wavelength anomalous diffraction using selenomethionine-labelled crystals and refined at 2.0 A resolution. Lp49 is composed of two domains and belongs to the all-beta-proteins class. The N-terminal domain folds in an immunoglobulin-like beta-sandwich structure, whereas the C-terminal domain presents a seven-bladed beta-propeller fold. Structural analysis of Lp49 indicates putative protein protein binding sites, suggesting a role in Leptospira-host interaction. This is the first crystal structure of a leptospiral antigen described to date. PMID- 18508282 TI - Grasping the diagonal: controlling attention to illusory stimuli for action and perception. AB - Since the pioneering work of [Aglioti, S., DeSouza, J. F., & Goodale, M. A. (1995). Size-contrast illusions deceive the eye but not the hand. Current Biology, 5(6), 679-685] visual illusions have been used to provide evidence for the functional division of labour within the visual system-one system for conscious perception and the other system for unconscious guidance of action. However, these studies were criticised for attentional mismatch between action and perception conditions and for the fact that grip size is not determined by the size of an object but also by surrounding obstacles. Stoettinger and Perner [Stoettinger, E., & Perner, J., (2006). Dissociating size representations for action and for conscious judgment: Grasping visual illusions without apparent obstacles. Consciousness and Cognition, 15, 269-284] used the diagonal illusion controlling for the influence of surrounding features on grip size and bimanual grasping to rule out attentional mismatch. Unfortunately, the latter objective was not fully achieved. In the present study, attentional mismatch was avoided by using only the dominant hand for action and for indicating perceived size. Results support the division of labour: Grip aperture follows actual size independent of illusory effects, while finger-thumb span indications of perceived length are clearly influenced by the illusion. PMID- 18508283 TI - Personality and mental time travel: a differential approach to autonoetic consciousness. AB - Recent research on autonoetic consciousness indicates that the ability to remember the past and the ability to project oneself into the future are closely related. The purpose of the present study was to confirm this proposition by examining whether the relationship observed between personality and episodic memory could be extended to episodic future thinking and, more generally, to investigate the influence of personality traits on self-information processing in the past and in the future. Results show that Neuroticism and Harm Avoidance predict more negative past memories and future projections. Other personality dimensions exhibit a more limited influence on mental time travel (MTT). Therefore, our study provide an additional evidence to the idea that MTT into the past and into the future rely on a common set of processes by which past experiences are used to envision the future. PMID- 18508284 TI - Pathology of the thoracic aorta: a morphologic review of 338 surgical specimens over a 7-year period. AB - A total of 338 surgical specimens of the thoracic aorta were studied morphologically. These were obtained from patients (238 males, 100 females) operated on for aortic aneurysm or dissection at the Careggi Hospital, Florence, Italy, between January 1999 and June 2005. Medial degeneration was diagnosed in 299 cases (138 aneurysms, 161 dissections), atherosclerosis with extensive medial destruction was seen in 32 cases (26 aneurysms, 6 dissections), and giant cell arteritis (GCA) was found in 7 cases (all aneurysms). These data show that medial degeneration was a common nonspecific histologic diagnosis in aortic resection after the occurrence of aneurysm or dissection. Considering that this diagnosis was made in as many as 118 patients over 70 years of age and in 175 patients over 65, normal advancing age would appear as the most frequent correlation with these aortic events. Ageing is also associated with the increasing number of cases of GCA with aortic involvement. Angiogenesis, always observed next to areas of medial damage whatever the underlying pathology, may contribute to the pathogenesis of aortic dissection and aneurysm. PMID- 18508285 TI - Hypertrophic cardiomyopathy presenting as restrictive cardiomyopathy: a case complicated by biventricular apical aneurysms and papillary fibroelastoma. AB - This report describes a 45-year-old male patient with nonobstructive hypertrophic cardiomyopathy, biventricular apical aneurysms, and papillary fibroelastoma. Histology revealed muscle fiber disarray, interstitial fibrosis, and mural vessel changes. The wall of the aneurysms showed fat infiltration, and loss and replacement of muscle fibers with fibrous tissues. A review of the literature suggests that this case is the first reported experience with the three pathologies occurring in combination. PMID- 18508286 TI - Up-regulation of a hydrogen peroxide-responsive pre-mRNA binding protein in atherosclerosis and intimal hyperplasia. AB - BACKGROUND: Multiple lines of investigation have implicated hydrogen peroxide (H(2)O(2)) as an important endogenous mediator of cell proliferation in the vessel wall. Heterogeneous nuclear ribonucleoprotein C (hnRNP-C), a nuclear pre mRNA binding protein that plays roles in vertebrate cell proliferation and differentiation, has been identified as a component of a vascular cell signaling pathway activated by low physiologic levels of H(2)O(2). The expression of hnRNP C in human arteries has not previously been assessed. METHODS: Segments of human proximal internal carotid arteries were evaluated for the expression of hnRNP-C by immunohistochemistry. RESULTS: In normal proximal internal carotid arteries, hnRNP-C is expressed predominantly by the endothelium, with significantly lower expression by medial smooth muscle. In preatherosclerotic intimal hyperplasia, hnRNP-C is up-regulated in the artery wall, due to the robust expression by the intimal smooth muscle cells, without up-regulation in the medial smooth muscle cells. In arteries with atherosclerotic lesions, there is strong expression of hnRNP-C not only by intimal cells but also by medial smooth muscle cells. CONCLUSIONS: The H(2)O(2) responsive pre-mRNA binding protein hnRNP-C is up regulated in atherosclerosis and in preatherosclerotic intimal hyperplasia in humans, supporting the hypothesis that H(2)O(2) is a regulator of vascular cell proliferation in these conditions. These data also suggest that hnRNP-C may be useful as a marker of vascular cell activation. PMID- 18508287 TI - Scientific review and recommendations on preclinical cardiovascular safety evaluation of biologics. AB - Biological therapeutic agents (biologicals), such as monoclonal antibodies (mAbs), are increasingly important in the treatment of human disease, and many types of biologicals are in clinical development. During preclinical drug development, cardiovascular safety pharmacology studies are performed to assess cardiac safety in accord with the ICH S7A and S7B regulations that guide these studies. The question arises, however, whether or not it is appropriate to apply these guidelines, which were devised primarily to standardize small molecule drug testing, to the cardiovascular evaluation of biologicals. We examined the scientific literature and formed a consensus of scientific opinion to determine if there is a rational basis for conducting an in vitro hERG assay as part of routine preclinical cardiovascular safety testing for biologicals. We conclude that mAb therapeutics have very low potential to interact with the extracellular or intracellular (pore) domains on hERG channel and, therefore, are highly unlikely to inhibit hERG channel activity based on their targeted, specific binding properties. Furthermore, mAb are large molecules (>140,000 Da) that cannot cross plasma membranes and therefore would be unable to access and block the promiscuous inner pore of the hERG channel, in contrast with typical small molecule drugs. Consequently, we recommend that it is not appropriate to conduct an in vitro hERG assay as part of a preclinical strategy for assessing the heart rate corrected QT interval (QTc) prolongation risk of mAbs and other types of biologicals. It is more appropriate to assess QTc risk by integrating cardiovascular endpoints into repeat-dose general toxicology studies performed in an appropriate non-rodent species. These recommendations should help shape future regulatory strategy and discussions for the cardiovascular safety pharmacology testing of mAbs as well as other biologicals and provide guidance for the preclinical cardiovascular evaluation of such agents. PMID- 18508288 TI - Advanced glycation end products in senile diabetic and nondiabetic patients with cataract. AB - BACKGROUND: Advanced glycation end products (AGE) have been reported to contribute to aging and cataract formation in the lens. In the present study, AGE immunoreactivity in human serum samples of normal senile subjects (n=31), senile diabetic patients without cataract (n=33), senile diabetic patients with cataract (n=30), senile nondiabetic with cataract (n=30), and normal young subjects (n=31) was investigated. METHODS: A noncompetitive ELISA with polyclonal anti-AGE antibody was performed. The patients were selected on clinical grounds from Eye Ward, Jinnah Postgraduate Medical Centre, Karachi, Pakistan. RESULTS: Fasting blood glucose, glycosylated hemoglobin, and serum fructosamine were estimated. Fasting blood glucose, HbA(1C), and serum fructosamine levels were significantly (P<.001) increased in senile diabetic patients with and without cataract as compared to nondiabetic senile patients with cataract and senile control subjects. However, the serum AGEs were found to be significantly (P<.001) increased in senile diabetic patients with cataract and senile nondiabetic patients with cataract followed by the diabetic patients without cataract as compared to senile control and young control subjects. In contrast to all four senile groups, the serum AGEs were significantly (P<.001) lower in young control subjects. CONCLUSIONS: The AGE distribution in the senile groups corroborates the hypothesis that the advanced glycation process might have a role in cataract formation, which in diabetic patients occurs vigorously as compared with nondiabetic cataract patients. PMID- 18508290 TI - Effects of satavaptan, a selective vasopressin V(2) receptor antagonist, on ascites and serum sodium in cirrhosis with hyponatremia: a randomized trial. AB - Hyponatremia in cirrhosis is associated with significant morbidity and mortality and complicates ascites management. Vasopressin receptor antagonists improve serum sodium concentration by increasing renal solute-free water excretion, but their effects on the management of ascites have not been assessed. Our aim was to investigate the effects of satavaptan, a highly selective vasopressin V(2) receptor antagonist, on ascites management and serum sodium in hyponatremic patients with cirrhosis. A total of 110 patients with cirrhosis, ascites, and hyponatremia (serum sodium < or =130 mmol/L) were included in a multicenter, double-blind, randomized, controlled study comparing three fixed doses of satavaptan (5 mg, 12.5 mg, or 25 mg once daily) versus placebo. Duration of treatment was 14 days and all patients received spironolactone at 100 mg/day. Satavaptan treatment was associated with improved control of ascites, as indicated by a reduction in body weight (mean change at Day 14 was +0.49 kg [+/ 4.99] for placebo versus +0.15 kg [+/-4.23], -1.59 kg [+/-4.60] and -1.68 kg [+/ 4.98] for the 5 mg, 12.5 mg, and 25 mg doses, respectively; P = 0.05 for a dose effect relationship overall) and a parallel reduction in abdominal girth. This beneficial effect on ascites was associated with improvements in serum sodium (mean change from baseline to day 5 was 1.3 +/- 4.2, 4.5 +/- 3.5, 4.5 +/- 4.8, and 6.6 +/- 4.3 mmol/L for the placebo group and the groups on satavaptan at 5 mg, 12.5 mg, and 25 mg/day, respectively; P < 0.01 for all compared to placebo). Thirst was significantly more common in patients treated with satavaptan compared to those treated with placebo, whereas the frequency of other adverse events was similar among groups. CONCLUSION: The V(2) receptor antagonist satavaptan improves the control of ascites and increases serum sodium in patients with cirrhosis, ascites, and hyponatremia under diuretic treatment. PMID- 18508291 TI - Host neutralizing responses and pathogenesis of hepatitis C virus infection. PMID- 18508292 TI - Diagnosis of hepatic nodules 20 mm or smaller in cirrhosis: prospective validation of the noninvasive diagnostic criteria for hepatocellular carcinoma. PMID- 18508295 TI - Interplay between anion-pi and hydrogen bonding interactions. AB - The interplay between two important noncovalent interactions involving aromatic rings is studied by means of high level ab initio calculations. They demonstrate that synergistic effects are present in complexes where anion-pi and hydrogen bonding interactions coexist. These synergistic effects have been studied using the "atoms-in-molecules" theory and the Molecular Interaction Potential with polarization partition scheme. The present study examines how these two interactions mutually influence each other. PMID- 18508296 TI - Rapid virological response and treatment duration for chronic hepatitis C genotype 1 patients: a randomized trial. AB - Recommended treatment for hepatitis C virus genotype 1 (HCV-1) patients is peginterferon plus ribavirin for 48 weeks. We assessed whether treatment duration of 24 weeks is as effective as standard treatment in HCV-1 patients with a rapid virological response (RVR; seronegative for hepatitis C virus [HCV] RNA at 4 weeks). Two hundred HCV-1 patients were randomized (1:1) to either 24 or 48 weeks of peginterferon-alpha-2a (180 microg/week) and ribavirin (1000-1200 mg/day) with a 24-week follow-up. The primary endpoint was a sustained virological response (SVR; seronegative for HCV RNA at 24-week follow-up). Overall, the 48-week arm had a significantly higher SVR rate (79%) than the 24-week arm (59%, P = 0.002). For 87 (43.5%) patients with an RVR, the 24-week arm had a lower SVR rate [88.9%; 95% confidence interval (CI): 80%-98%] than the 48-week arm (100%, P = 0.056). For 52 patients with low baseline viremia (<400,000 IU/mL) and an RVR, the 24 week arm had rates (CI) of relapse and SVR of 3.6% (-3%-11%) and 96.4% (89% 103%), respectively, which were comparable to those of the 48-week arm (0% and 100%) with difference (CI) of 3.6% (-7.2%-6.6%) and -3.6% (-14.3% to -0.6%), respectively. Multivariate analysis in all patients showed that RVR was the strongest independent factor associated with an SVR, followed by treatment duration, mean weight-based exposure of ribavirin, and baseline viral load. CONCLUSION: HCV-1 patients derive a significantly better SVR from 48 weeks versus 24 weeks of peginterferon/ribavirin even if they attain an RVR. Both 24 and 48 weeks of therapy can achieve high SVR rates (>96%) in HCV-1 patients with low viral loads and an RVR. PMID- 18508297 TI - Searching high and low: cancer stem cells in the eye. PMID- 18508298 TI - Insulin resistance in chronic hepatitis C, genotypes 1 and 4: the unfortunate reality. PMID- 18508299 TI - Hepatocellular carcinoma development requires hepatic stem cells with altered transforming growth factor and interleukin-6 signaling. PMID- 18508302 TI - Transient elastography, liver stiffness values, and acute hepatopathy. PMID- 18508304 TI - A grapefruit a day for patients infected with hepatitis C? PMID- 18508305 TI - Molecular adsorbents recirculating system (MARS) and the failing liver: a negative editorial for a positive trial? PMID- 18508306 TI - Biopsy diagnosis of hepatocellular carcinoma. PMID- 18508307 TI - On relativistic effects in ground state potential curves of Zn2, Cd2, and Hg2 dimers. A CCSD(T) study. AB - The ground state potential curves of the Zn2, Cd2, and Hg2 dimers calculated at different levels of theory are presented and compared with each other as well as with experimental and other theoretical studies. The calculations at the level of Dirac-Coulomb Hamiltonian (DCH), 4-component spin-free Hamiltonian, nonrelativistic Levy-Leblond Hamiltonian and at the level of simple Coulombic correction to DCH are presented. The potential curves are calculated in an all electron supermolecular approach including the correction to basis set superposition error (BSSE). Electron correlation is treated at the coupled cluster level including single and double excitations and noniterative triple corrections, CCSD(T). In addition, simulations of the temperature dependence of dynamic viscosities in the low-density limit using the obtained ground state potential curves are presented. PMID- 18508308 TI - End-stage chronic liver disease: time to define a good death. PMID- 18508309 TI - Workforce issues in hepatology: what is needed? PMID- 18508312 TI - What is idiosyncratic hepatotoxicity? What is it not? PMID- 18508313 TI - Single-nucleotide polymorphism-mass array reveals commonly deleted regions at 3p22 and 3p14.2 associate with poor clinical outcome in esophageal squamous cell carcinoma. AB - Esophageal squamous cell carcinoma (ESCC) is one of the most common solid tumors in the world with poor prognosis. Deletion of chromosome 3p is one of the most frequent chromosomal alterations in ESCC, suggesting the existence of one or more tumor suppressor genes (TSGs) at this region. In the present study, a recently developed high-throughput and high-resolution technology, single-nucleotide polymorphism (SNP)-mass array, was applied to investigate loss of heterozygosity on 3p in 100 primary ESCC cases with 386 SNP markers. Four commonly deleted regions (CDRs) at 3p26.3, 3p22, 3p21.3 and 3p14.2 were identified. Absent and down-regulated expression of several candidate TSGs, including CHL1, PCAF, RBMS3, PLCD1 and CACNA2D3, were detected in primary ESCC tumors and ESCC cell lines. Moreover, deletions of CDRs 2 and 4 were correlated with advanced tumor stage and deletion of CDR2 was associated with tumor metastasis in ESCC. Our findings provided evidence that minimal deleted regions at 3p26.3, 3p22, 3p21.3 and 3p14.2 containing potential TSGs may contribute to the pathogenesis of esophageal cancer. PMID- 18508314 TI - Risk of gastric cancer in asymptomatic, middle-aged Japanese subjects based on serum pepsinogen and Helicobacter pylori antibody levels. AB - A total of 5,209 asymptomatic, middle-aged subjects, whose serum pepsinogen (PG) and Helicobacter pylori antibody levels had been assessed, were followed for 10 years. Subjects with positive serum H. pylori antibodies (>50 U/mL) had an increased cancer risk (HR = 3.48, 95% CI = 1.26-9.64). Risk of gastric cancer increased as the antibody level increased; the H. pylori-positive group with antibody levels >500 U/mL had the highest incidence rate (325/100,000 person years). Cancer development also increased with a reduced serum PG I level or a reduced PG I/II ratio; the risk was significantly elevated with serum PG I level or=30 ng/mL (HR = 3.81, 95% CI = 1.10-13.21). Using H. pylori antibody and PG levels, subgroups with an especially high or low cancer incidence rate could be identified. H. pylori-negative or indeterminate subjects with low PG level (PG I 500 U/mL and a low PG level were among the subgroups with a high cancer incidence rate (over 400/100,000 person-years). In contrast, H. pylori negative subjects with a PG I level >70 ng/mL or a PG I/II ratio >3.0 had the lowest risk; none of these subjects developed cancer. Thus, serum PG levels and/or H. pylori antibody levels can be used to predict the risk of cancer in individuals with H. pylori-related gastritis from the general population. PMID- 18508315 TI - PKC alpha protein but not kinase activity is critical for glioma cell proliferation and survival. AB - Protein kinase C alpha (PKCalpha) has been implicated in tumor development with high levels of PKCalpha expression being associated with various malignancies including glioblastomas and tumors of the breast and prostate. To account for its upregulation in these cancers, studies have suggested that PKCalpha plays a role in promoting cell survival. Here we show by siRNA depletion in U87MG glioma cells that a critical threshold level of PKCalpha protein expression is essential for their growth in the presence of serum and for their survival following serum deprivation. Derivation of PKCalpha wt and KO mouse embryo fibroblast cell lines confirms a role for PKCalpha in protecting cells from apoptosis induced by serum deprivation. Notably, PKCalpha was found to mediate chemo-protection in these fibroblastic cell lines. In U87MG cells PKCalpha does not confer chemoprotection though this likely reflects growth arrest associated with its depletion. To determine the requirements for catalytic function, comparison was made between distinct classes of PKC inhibitors. In contrast to loss of PKCalpha protein, inhibition of PKC kinase activity in glioma cell lines does not significantly inhibit growth or survival. Conversely, inhibition with calphostin C, which targets the regulatory domain of PKC, potently inhibits proliferation and induces apoptosis. Evidence is presented that it is the fully phosphorylated, folded form of PKCalpha that confers this activity-independent behaviour. These results indicate an essential pro-proliferative and pro-survival role for PKCalpha in glioma but question the use of ATP competitive inhibitors as therapeutics, either alone, or in combination with chemotoxic agents. PMID- 18508316 TI - Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells. AB - Transforming growth factor-beta (TGF-beta) plays a dual and complex role in human cancer. In this report, we observe a specific set of MicroRNAs (miRNAs) changed in response to TGF-beta in human hepatocellular carcinoma (HCC) cells by miRNA microarray screening. A cluster of miRNA, miR-23a approximately 27a approximately 24, is induced in an early stage by TGF-beta in Huh-7 cells. Knockdown of Smad4, Smad2 or Smad3 expression by RNA interference can attenuate the response of miR 23a approximately 27a approximately 24 to TGF-beta addition, indicating that this induction is dependent on Smad pathway. We also explore that miR-23a approximately 27a approximately 24 can function as an antiapoptotic and proliferation-promoting factor in liver cancer cells. In addition, expression of this miRNA cluster is found to be remarkably upregulated in HCC tissues versus normal liver tissues. These findings suggest a novel, alternative mechanism through which TGF-beta could induce specific miRNA expression to escape from tumor-suppressive response in HCC cells. PMID- 18508317 TI - Enhanced expression of NADPH oxidase Nox4 in human gliomas and its roles in cell proliferation and survival. AB - Reactive oxygen species (ROS) have been attracting attention as mediators of various cell-signaling pathways. Nox-family NADPH oxidases have proven to be a major source of ROS production in various cell types and have crucial roles in various physiological and pathological processes. In this study, we show that Nox4, a member of Nox family, is prominently expressed in various neuroepithelial tumors by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical studies. We quantified Nox4 mRNA expression by real-time PCR in tumor specimens from 58 patients with astrocytomas and found that the expression levels of Nox4 mRNA in glioblastomas (WHO grade IV) were significantly higher than those in other astrocytomas (WHO grade II and III). In addition, we show that specific knockdown of Nox4 expression by RNA interference results in cell-growth inhibition and enhances induction of apoptosis by chemotherapeutic agents, such as cisplatin, in cultured glioma cell lines. Based on these observations, enhanced expression of Nox4 appears to be involved in cell proliferation and survival in glioma cells. PMID- 18508318 TI - Potential pathogenic role of Mycobacterium avium subspecies paratuberculosis in Crohn's disease. PMID- 18508319 TI - Complete remission in a case of severe multi-resistant idiopathic thrombocytopenic purpura after Helicobacter pylori eradication. PMID- 18508320 TI - Impact of genetic variation on perioperative bleeding. AB - Variation in bleeding in the perioperative period is a complex and multifactorial event associated with immediate and delayed consequences for the patient and health care resources. Little is known about the complex genetic influences on perioperative bleeding. With the discovery of multiple variations in the human genome and ever-growing databases of well-phenotyped surgical patients, better identification of patients at risk of bleeding is becoming a reality. In this review, polymorphisms in the platelet receptor genes, plasminogen activator inhibitor, and angiotensin genes among others will be discussed. We will explore the nature, effects, and implications of the genetics that influence perioperative bleeding above and beyond surgical bleeding, particularly in cardiac surgery. PMID- 18508321 TI - Reduced dose of lenograstim is as efficacious as standard dose of filgrastim for peripheral blood stem cell mobilization and transplantation: a randomized study in patients undergoing autologous peripheral stem cell transplantation. AB - In vitro studies have demonstrated a 27% increased efficacy of lenograstim over filgrastim. However, equal doses of 10 microg/kg/day of filgrastim and lenograstim have been recommended for mobilization of CD34+ cells without associated chemotherapy. In this study, we investigated whether a 25% reduced dose of lenograstim at 7.5 microg/kg/day is equavalent to 10 microg/kg/day filgrastim for autologous peripheral blood stem cell (PBSC) mobilization and transplantation. A total of 40 consecutive patients were randomized to either filgrastim (n = 20) or lenograstim (n = 20). The two cohorts were similar in regard to disease, sex, body weight, body surface area, conditioning regimens, previous chemotherapy cycles and radiotherapy. Each growth factor was administered for 4 consecutive days. The first PBSC apheresis was done on the 5th day. In the posttransplant period, the same G-CSF was given at 5 microg/kg/day until leukocyte engraftment. Successful mobilization was achieved in 95% of patients. Successful mobilization with the first apheresis, was achieved in 10/20 (50%) patients in the filgrastim group versus 9/20 (46%) patients in the lenograstim group. No significant difference was seen in the median number of CD34+cells mobilized, as well as the median number of apheresis, median volume of apheresis, percentage of CD34+ cells, and CD34+ cell number. Leukocyte and platelet engraftments, the number of days requiring G-CSF and parenteral antibiotics, the number of transfusions were similar in both groups in the posttransplant period. Lenograstim 7.5 microg/kg/day is as efficious as filgrastim 10 microg/kg/day for autologous PBSC mobilization and transplantation. PMID- 18508322 TI - Attenuated doses of rituximab for the treatment of adults with autoimmune cytopenias. PMID- 18508323 TI - Delayed hemolysis after intravenous anti-D immune globulin infusion in a patient with idiopathic thrombocytopenic purpura. PMID- 18508324 TI - Recent advances in affinity capillary electrophoresis (2007). AB - Papers detailing the use of affinity capillary electrophoresis (ACE) in examining binding parameters between biological species are summarized in this review. The works cited in this review were published during 2007 and were drawn from the major chemical and biochemical journals and especially the major analytical chemistry literature. PMID- 18508325 TI - Psoas muscle cysticercosis presenting as acute appendicitis. PMID- 18508326 TI - Blood doping in athletes--detection of allogeneic blood transfusions by flow cytofluorometry. AB - Athletes may undergo blood transfusion to increase their red cell mass and the oxygen carrying capacity of their blood in order to confer a competitive advantage. Allogeneic transfusions are normally mismatched at one or more minor blood group antigens. The most sensitive and accurate method known to detect this form of blood doping is flow cytometry. Low percentages of antigen-positive and antigen-negative red blood cells (RBCs) can be quantitated using suitable specific alloantibodies and careful analysis. By testing blood samples taken at various times, a reduction in the percentage of a minor population of RBCs will indicate transfusion has occurred. PMID- 18508327 TI - Conformation and absolute configuration of 2-naphthylalkylsulfoxides by combined use of dynamic NMR, ECD spectroscopy, DFT computations, and X-ray diffraction. AB - The E/Z conformer ratios of five 2-naphtylalkylsulfoxides were determined by means of dynamic NMR spectroscopy at very low temperatures and this information was used to predict, by means of DFT calculations, the ECD spectra of the R and S enantiomers. The latter were separated by enantioselective HPLC technique and the comparison of theoretical and experimental ECD spectra allowed the absolute configurations to be determined. In the case of 2-naphtyl tert-butylsulfoxide (1), the assignment was independently confirmed by anomalous dispersion using single crystal X-ray crystallography. PMID- 18508328 TI - A DNA transporter based on mesoporous silica nanospheres mediated with polycation poly(allylamine hydrochloride) coating on mesopore surface. AB - Mesoporous silica nanospheres (MPNSs) were synthesized by the sol-gel method and were investigated by X-ray diffraction, nitrogen sorption, and transmission electron microscopy. The analyses showed that the MPNSs were highly dispersed, with a diameter range of 50-250 nm. Further, the polycation poly(allylamine hydrochloride) was coated on silica MPNSs to form a positively charged surface and to facilitate the loading of DNA molecules. The composite was successfully used as a intercellular carrier of DNA molecules, and this was confirmed by agarose gel electrophoresis and confocal laser scanning microscopy. Evidently, this composite has potential to be used as a carrier for a DNA delivery system. PMID- 18508329 TI - PAX3/7 expression coincides with MyoD during chronic skeletal muscle overload. AB - Paired box (Pax) proteins 3 and 7 are key determinants for embryonic skeletal muscle development by initiating myogenic regulatory factor (MRF) gene expression. We show that Pax3 and 7 participate in adult skeletal muscle plasticity during the initial responses to chronic overload (< or =7 days) and appear to coordinate MyoD expression, a member of the MRF family of genes. Pax3 and 7 mRNA were higher than control within 12 h after initiation of overload, preceded the increase in MyoD mRNA on day 1, and peaked on day 2. On days 3 and 7, Pax7 mRNA remained higher than control, suggesting that satellite cell self renewal was occurring. Pax3 and 7 and MyoD protein levels were higher than control on days 2 and 3. These data indicate that Pax3 and 7 coordinate the recapitulation of developmental-like regulatory mechanisms in response to growth inducing stimuli in adult skeletal muscle, presumably through activation of satellite cells. PMID- 18508330 TI - A defence of rigorous method in clinical science. PMID- 18508331 TI - Satisfaction with urethral injection of botulinum toxin A for detrusor sphincter dyssynergia in patients with spinal cord lesion. AB - AIMS: To investigate satisfaction with urination quality of life (QoL) after treatment with urethral sphincter botulinum toxin A (BoNT-A) injection for difficult urination in patients with spinal cord lesions and detrusor sphincter dyssynergia (DSD). METHODS: Thirty-three patients with spinal cord lesion and DSD were treated with transurethral sphincter injection of 100 U of BoNT-A (BOTOX) for the main symptom of difficult urination. The urodynamic parameters, QoL scores obtained using UDI-6 and IIQ-7 and general satisfaction and dissatisfaction were assessed. RESULTS: An overall satisfactory result was perceived by 60.6% of patients. Urodynamic parameters showed significant improvement in voiding detrusor pressure (45.7 +/- 22.7 vs. 30.7 +/- 15.5 cmH(2)O, P = 0.016), maximum flow rate (6.8 +/- 5.7 vs. 9.2 +/- 7.7 ml/sec, P = 0.047) and postvoid residual volume (PVR, 160 +/- 124 vs. 75 +/- 105 ml, P = 0.025). IIQ-7 scores showed significant improvement (19.0 +/- 2.4 vs. 17.1 +/- 2.9, P = 0.001) but not in UDI-6 scores (8.6 +/- 2.7 vs. 7.9 +/- 3.7, P = 0.252). Less difficult urination and less PVR needing clean intermittent catheterization were the major reasons for satisfaction, whereas increase in incontinence grade was the major reason for dissatisfaction in patients receiving urethral BoNT-A injection. CONCLUSION: This study found that urethral sphincter BoNT-A injection for DSD does not prove convincing results in all patients and should be done only exceptionally in well-chosen patients with spinal cord lesions. Dissatisfaction in the patients was mainly due to an increased incontinence grade that was not anticipated before urethral BoNT-A injection. PMID- 18508332 TI - Management of neurogenic bladder patients in The Netherlands: do urologists follow guidelines? AB - AIMS: Preventive measurements are obligatory for optimal treatment of neurogenic bladder patients. We investigated Dutch urological practice in neurogenic bladder patients in relation to the available guidelines on neurogenic bladder. METHODS: A 12-point survey was sent to all 304 certified urologists of the Dutch Urological Association in July 2007. Management of urinary tract infections (UTI), follow-up of the lower and upper urinary tract and treatment techniques were topics of inquiry. RESULTS: The response rate was 46% of which 94% were involved in treatment of neurogenic bladder patients. Follow-up was performed every 6 or 12 months in 86%, urine analysis respectively in 85% and upper urinary tract imaging with ultrasound in 60%; only 12% routinely repeated urodynamics. Uncomplicated UTI was treated for 7.1 days in women and 9.2 days in males. Eighteen percent used the EAU guidelines on neurogenic bladder frequently, 35% did occasionally and 47% did not use them at all. Urologists, that used these guidelines frequently, treated asymptomatic UTIs significantly less frequent in patients on intermittent catheterization (5% vs. 25%; P = 0.008); no difference was found in frequency of follow-up, urine analysis, surveillance of upper urinary tract, use of routine urodynamics, and availability of treatment techniques. Video urodynamics, the golden standard, was used by only 11% of the adult urologists. CONCLUSION: The majority of Dutch urologists is involved in the treatment of neurogenic bladder, which suggests a less prominent role for specialized centers. Furthermore, urodynamics are not performed routinely. These observations are contrary to the recommendations of the available guidelines. PMID- 18508333 TI - Modeling the clinical assessment of men with suspected obstructed voiding using Bayes' Theorem. AB - AIMS: Pressure-flow studies (PFS) are the only reliable way to diagnose bladder outlet obstruction (BOO) in men with lower urinary tract symptoms (LUTS). However, in routine clinical practice, BOO is usually inferred by any of a number of tests (symptoms, flow rate, prostate size...). Bayes' Theorem provides a mathematical method, which may be similar to the process used by clinicians, for combining the results of multiple tests to reach a diagnosis. We have applied Bayes' Theorem to the results of several tests known weakly to predict BOO in men with LUTS to assess if they improve the diagnostic accuracy of a flow rate test which alone is known to predict obstruction moderately well. METHODS: We applied Bayes' Theorem to data from 50 patients using Q(max) alone and with the inclusion of additional variables (IPSS, PSA, and residual urine), to establish individual probabilities of BOO. The chi-squared statistic (with trend) was used to compare the relative diagnostic values, against the BOO index calculated from the results of subsequent PFS. RESULTS: The diagnostic value of Q(max) alone (chi-squared = 9.2, P = 0.002), was superior than that for the Bayesian model using the combination of tests available (chi-squared = 4.9, P = 0.026). CONCLUSIONS: Although in our sample relevant additional tests do not improve the diagnostic power of Q(max) as a predictor of BOO, we believe the Bayesian approach is conceptually suited to modeling clinical decision making but may be better tested for a more clinically relevant outcome such as treatment response. PMID- 18508334 TI - Neurogenic detrusor overactivity: comparison between complete and incomplete spinal cord injury patients. AB - AIMS: To compare leak-point intravesical pressure and cystometric capacity in complete and incomplete spinal cord injury (SCI) patients with neurogenic detrusor overactivity (NDO). METHODS: Retrospective study of filling cystometry at non-physiological filling rate in 80 SCI patients at rehabilitation or annual check-up using Dantec Etude urodynamic machine. RESULTS: Fifty neurologically complete (ASIA (American Spinal Injury Association) A) and 30 incomplete (ASIA B E) were diagnosed with neurogenic detrusor overactivity, all with suprasacral level of injury. Mean Pves leak-point pressure (Pves LPP) at cystometric capacity for ASIA A group was 79 +/- 30 cmH(2)O (range 26-140) and mean Pves LPP for ASIA B-E group was 70 +/- 29 cmH(2)O (range 25-130). There was no significant difference between groups (P = 0.234). Mean CC (cystometric capacity) for ASIA A group was 239 +/- 107 ml (range 47-526) and mean CC for ASIA B-E group was 227 +/ 125 ml (range 42-500). Again, no significant difference was found (P = 0.655). CONCLUSIONS: No difference in cystometric capacity and intravesical leak point pressure at terminal detrusor overactivity was shown between complete and incomplete spinal cord injury patients in our survey, that is, represented findings are equally unfavorable for both groups. Incomplete SCI patients with NDO should be tested with cystometry and observed with same caution as we proceed in complete SCI patients. PMID- 18508335 TI - Re: Results on function and safety of the Safyre-t, a hybrid transobturator vaginal sling for the treatment of stress urodynamic incontinence, Epub ahead, Neurourol Urodyn 2007. PMID- 18508336 TI - Novel "anchor modification" of polymeric biomaterial surfaces by the utilization of cyclodextrin inclusion complex supramolecules. AB - In this article, a novel approach for the surface modification of polymeric biomaterials by the utilization of supramolecules was studied. The supramolecules selected were cyclodextrin inclusion complexes (CICs). The biomaterial selected for surface modification was plasticized poly (vinyl chloride) (PVC-P). Results indicate that when the CICs were blended into PVC-P, they tend to migrate and "anchor" on the surface to achieve a remarkable protein-resistant surface, with improved blood compatibility. In comparison with a physical mixture of cyclodextrins and a "guest" molecule, such as poly(ethylene oxide) (PEO) poly(propylene oxide) (PPO)-PEO and PPO-PEO-PPO for PVC-P modification, CICs modified PVC-P are more consistent in processing and achieve reproducible surface characteristics. Based on this study, a novel "anchor modification" was proposed regarding CICs modified surface. This "anchor modification" is likely to reduce plasticizer extraction from PVC-P and also can be utilized for the modification of polymers other than PVC-P. PMID- 18508337 TI - Biocompatibility of total joint replacements: A review. AB - Total joint replacement is one of the most clinically successful, cost-effective surgical procedures. These operations have been shown to improve pain, function and mobility in patients with end-stage arthritis. However, joint replacements that will allow full, unrestricted, high impact activities and last the patient's lifetime have not yet been devised. This is due to biocompatibility issues related to material science, biomechanics, and host responses. In this review, three issues that are highly pertinent to biocompatibility of joint replacements are examined. These topics include how implants initially osseointegrate within bone, potential adverse effects of byproducts of wear that can lead to aseptic loosening and periprosthetic osteolysis, and the potential for new bearing surfaces to extend the lifetime of implants. A clear understanding of these important issues will facilitate the development of novel strategies to improve the longevity and function of implants for joint replacement in the future. PMID- 18508338 TI - Protein recognition onto silica particles using chitosan as intermedium substrate. AB - A novel molecular imprinting method was used to prepare twice-coated silica particles with specific recognition sites for hemoglobin. Chitosan was used as an intermedium to be coated on silica particles via phase inversion process, and the abundance of exposed amine groups (NH2) were active sites for introducing aldehyde groups. After hemoglobin was covalently immobilized by forming imine bonds with the aldehyde groups, acrylamide was then polymerized onto chitosan coated silica particles to form the recognition sites. The obtained hemoglobin imprinted [molecularly imprinted polymer (MIP)] beads were characterized by FTIR spectroscopy, thermogravimetric analysis (TG), and scanning electron microscopy (SEM). The MIP particles exhibited selectively adsorption for the imprinted protein compared to the nonselectively adsorption for most of proteins of the nonimprinted (NIP) beads. PMID- 18508339 TI - Novel poly(HEMA-co-METAC)/alginate semi-interpenetrating hydrogels for biomedical applications: synthesis and characterization. AB - Novel p(HEMA-co-METAC)/alginate semi-interpenetrating hydrogels (semi-IPNs) were developed in the attempt to improve poly(hydroxyethylmethacrylate) biological response, extending its applications in the biomedical field. Materials with different alginate contents were synthesized by copolymerization of 2 hydroxyethylmethacrylate and 2-methacryloxy ethyltrimethyl ammonium chloride monomers in the presence of aqueous solutions of alginate and characterized with respect to p(HEMA-co-METAC) synthesized in the presence of water. Swelling studies in water revealed high values of water uptake (>100%) with marked differences in the swelling degree at increasing polysaccharide content. The effect of ionic strength and of pH on the swelling behavior of hydrogels was also investigated. Higher ionic strengths resulted in a minor swelling degree accordingly with hydrogels polyelectrolyte nature. The introduction of the natural ionizable polysaccharide into the network made the semi-IPNs swelling depending on pH values of the solutions. A biological characterization was performed in terms of protein absorption on hydrogel surfaces, cytotoxicity (ISO 10993-5) and cell adhesion and proliferation studies using both murine 3T3 and human fibroblasts. Hydrogels proved noncytotoxic; moreover, semi-IPN surfaces allowed cell attachment and proliferation, thus supporting their potential biomedical use. PMID- 18508340 TI - Adolescent spinal muscular atrophy with calf hypertrophy and a deletion in the SMN gene. AB - Spinal muscular atrophy (SMA) is generally associated with proximal weakness and muscle wasting. An X-linked variant with calf hypertrophy has been reported. We describe a young man with SMA type 4 with prominent calf hypertrophy in whom DNA analysis showed a homozygous deletion of exons 7 and 8 in the telomeric copy of the survival motor neuron gene. Calf hypertrophy may be seen uncommonly in autosomally inherited SMA. PMID- 18508341 TI - Analysis of double discharges in amyotrophic lateral sclerosis. AB - Double discharges of motor units (MUs) occurring during sustained voluntary muscle contractions are observed occasionally in healthy muscles and more frequently in disorders of the neuromuscular system. In healthy subjects, double discharges are generated in motoneurons (MNs) and are considered to be a sign of their increased excitability. Therefore, an analysis of their firing pattern may provide information on the state of MNs in neuromuscular diseases, particularly in amyotrophic lateral sclerosis (ALS), whose etiology remains to be disclosed. Firing patterns of MUs capable of firing double discharges were analyzed in brachial biceps of 14 patients with ALS (184 MUs) and 8 healthy control subjects (102 MUs). The incidence of MUs capable of firing double discharges was significantly higher in ALS patients (28.8%) than in controls (3.9%). The majority of doublet interval durations (range 4-8 ms) as well as firing patterns of doubling MUs did not differ between subject groups. Although our data confirm the hyperexcitability of the MN pool in ALS, analysis of firing characteristics of doubling MUs indicates that doublet generation is governed by the same mechanism as in controls, that is, by delayed depolarization. Our findings may provide insight into MN function in ALS. PMID- 18508342 TI - Frequency of cyclic lumbar loading is a risk factor for cumulative trauma disorder. AB - Epidemiologic studies indicate that repetitive (cyclic) occupational activities lead to a cumulative trauma disorder (CTD), and the frequency or velocity of the movement is one of the risk factors. Experimental neurophysiological evidence to confirm the epidemiology is not available. The response of the multifidus muscles to cyclic loading in anterior lumbar flexion-extension was assessed to test the hypothesis that high-frequency loading may induce an acute neuromuscular disorder leading to CTD. Two groups of feline preparations were subjected to cyclic loading with a peak of 20 N: one at 0.25 HZ and the second at 0.5 HZ, with an equal number of cycles. Electromyogram (EMG), lumbar displacement and load were recorded throughout the loading periods and during single-cycle tests over a 7 hour rest period following the load-rest sessions. A model was developed to quantify the creep and neuromuscular responses, and analysis of variance (ANOVA) was applied to assess significance of the results. The group exposed to 0.5 HZ exhibited spontaneous spasms followed by sustained spasms during the loading periods. During the 7-hour recovery period, a significant (P < 0.001) delayed hyperexcitability as well as sustained spasms of the multifidi were present in the last 5 hours, confirming a significant (P < 0.024 to P < 0.042) acute neuromuscular disorder. High-frequency cyclic loading of the lumbar spine may trigger a severe acute neuromuscular disorder, as evidenced by the sustained spasms and delayed hyperexcitability, and should be considered as a risk factor. We suggest that workers avoid high-frequency exposure to cyclic activity in order to prevent the development of cumulative trauma disorder. PMID- 18508343 TI - Chronic inflammatory demyelinating polyneuropathy-like disorder associated with amyotrophic lateral sclerosis. AB - The association between demyelinating neuropathy and amyotrophic lateral sclerosis (ALS) has been reported rarely. We report four patients who presented with clinical features and investigations suggestive of a neuropathy but who later progressed and received a final diagnosis of ALS according to the original El Escorial criteria. Three patients met the European Federation of Neurological Societies/Peripheral Nerve Society (EFNS/PNS) 2006 definition for "definite" chronic inflammatory demyelinating polyneuropathy (CIDP), as well as the American Academy of Neurology (AAN) 1991 and Nicolas et al. [2002] electrodiagnostic criteria for CIDP. Two of them showed segmental demyelination on teased-fiber preparations and one had a raised cerebrospinal fluid protein level. Another patient met the Nicolas et al. CIDP criteria and presented with brachial plexus hyperintensity on T2-weighted magnetic resonance imaging, indicative of an inflammatory process. No significant objective response to immunomodulatory treatment was observed in any of these patients who all subsequently progressed unfavorably. PMID- 18508344 TI - Hormonal and nutritional control of L-carnitine uptake in myoblastic C2C12 cells. AB - L-Carnitine plays an important role in skeletal muscle bioenergetics, and its bioavailability and thus its import may be crucial for muscle function. We studied the effect of thyroid hormone, insulin, and iron overload, hormones and nutrients known to alter muscle metabolism, on L-carnitine import into C2C12 cells. We report here L-carnitine uptake is increased by thyroid hormones and decreased by iron. Insulin was found to be ineffective in altering the L carnitine uptake. PMID- 18508345 TI - Chronic sensorimotor polyradiculopathy with antibodies to P2: an electrophysiological and immunoproteomic analysis. AB - In this study we report a patient with chronic progressive sensory ataxia, proximal weakness, immunoglobulin M (IgM) monoclonal gammopathy, and elevated protein levels in the cerebrospinal fluid, who showed a good response to prednisone. Electrophysiological study disclosed abnormalities predominantly of late responses (F waves and H reflexes), with no evidence of demyelination in the peripheral nerves, suggesting motor and preganglionic sensory nerve roots as the site of the lesion. An immune-mediated pathogenesis was considered and, to identify possible target antigens, we performed bidimensional electrophoresis and a Western blot study. Based on the suspected lesion site, we used human anterior and posterior root extracts. We identified IgM reactivity against peripheral nerve myelin protein P2. Enzyme-linked immunosorbent assay confirmed IgM reactivity toward one synthetic peptide from P2. To our knowledge, reactivity against P2 has not been reported previously in a paraproteinemic neuropathy. Furthermore, we demonstrated that bidimensional electrophoresis and Western blot of the tissue involved, as determined by clinical and electrophysiological studies, may be useful to establish clinical-immunological correlations in paraproteinemic neuropathies. PMID- 18508346 TI - Nine-year case history of monofocal motor neuropathy. AB - A 36-year-old man had a 6-year history of progressive left hand and finger extension weakness. Extensive electrophysiological studies showed only a partial motor conduction block along the left radial nerve at the mid-arm. Several intravenous immunoglobulin treatments were clinically successful. A 3-year follow up did not show any clinical or electrophysiological involvement of other nerves. A monofocal motor neuropathy, as a variant of multifocal motor neuropathy, was diagnosed. PMID- 18508347 TI - Ross syndrome associated with cytomegalovirus infection. AB - We report a 40-year-old woman who developed Ross syndrome (impairment of sweating and thermoregulation, tonic pupils, and hyporeflexia) associated with cytomegalovirus (CMV) infection. Her serum CMV IgM and IgG antibody titer levels were elevated. Along with clinical improvement, a gradual decrease of her elevated CMV IgM antibody titer level was seen with a continued increase in her CMV IgG antibody titer level. The CMV IgM antibody titer was also positive in the cerebrospinal fluid. PMID- 18508348 TI - Obturator mononeuropathy caused by lipomatosis of the nerve: a case report. AB - We report a patient who presented with the clinical features of obturator mononeuropathy. Abdomino-pelvic computed tomography revealed a fusiform mass in the right perivesical space; magnetic resonance imaging (MRI) showed characteristic "coaxial-cable-like" appearance in cross-section and "spaghetti like" appearance in longitudinal section, pathognomonic of lipomatosis of the nerve. Nerve lipomatosis as the cause of obturator neuropathy has not been previously reported. MRI provides definite and graphic proof of the diagnosis. PMID- 18508350 TI - Reference data for quantitative motor unit potential analysis in the genioglossus muscle. PMID- 18508349 TI - Glial fibrillary acidic protein: a marker of axonal Guillain-Barre syndrome and outcome. AB - Glial fibrillary acid protein (GFAP) is increased in serum and cerebrospinal fluid of patients with dementia, traumatic brain injury, stroke, and multiple sclerosis. To determine whether GFAP is increased in Guillain-Barre syndrome (GBS) we evaluated serum GFAP in 30 controls, 20 patients with acute inflammatory demyelinating neuropathy (AIDP), and 17 with primary axonal GBS. Serum GFAP levels were increased in axonal GBS (median, 0.74) compared with controls (median, 0.41; P < 0.0001) and AIDP (median, 0.58; P = 0.0015). GFAP levels correlated with Hughes grades (serum r = 0.74; P < 0.0001) 6 months after neuropathy onset. Applying the cutoff value in serum of 0.63 to the diagnosis of axonal GBS, we obtained a sensitivity of 76.5% and a specificity of 86%. Thus, serum GFAP levels may be used in GBS as a diagnostic marker of the axonal variant and to predict outcome. PMID- 18508351 TI - Effect of small motor unit potentials on the motor unit number estimate. AB - Small surface motor unit potentials (S-MUPs) may have a negative influence on the variability of the motor unit number estimate (MUNE). According to published consensus criteria S-MUPs with a negative peak amplitude smaller than 10 muV should be omitted. The effect of omitting small S-MUPs on the MUNE was evaluated using a simulation model. The model incorporated a healthy and amyotrophic lateral sclerosis (ALS) distribution formed with real S-MUPs. Using a random drawing process the MUNE was calculated with and without small S-MUPs. In the healthy population 27% of all S-MUPs were small. MUNE determined without these S MUPs was marginally less variable. However, MUNE values dropped about 24% at a sample size of 20. In ALS, only 12% of the total population of 130 S-MUPs were small. MUNE dropped about 12% without the small S-MUPs. By omitting small S-MUPs the differences between the healthy and ALS distributions become smaller. Therefore, incorporating small S-MUPs in the estimate is suggested. PMID- 18508352 TI - Physico/chemical characterization and in vivo evaluation of nanothickness bioceramic depositions on alumina-blasted/acid-etched Ti-6Al-4V implant surfaces. AB - The objective of this study was to physico/chemically characterize and evaluate the in vivo performance of two nanothickness ion beam assisted depositions (IBAD) of bioceramic coatings on implants in a beagle model. Alumina-blasted/acid-etched (AB/AE) Ti-6Al-4V implants were subjected to two different IBAD depositions (IBAD I and IBAD II), which were physico/chemically characterized by SEM, EDS, XPS, XPS + ion-beam milling (depth profiling), XRD, AFM, and ToF-SIMS. A beagle dog tibia model was utilized for histomorphometric and biomechanical (torque) comparison between AB/AE, IBAD I, IBAD II, and plasma-sprayed hydroxyapatite (PSHA) coated implants that remained in vivo for 3 and 5 weeks. The coatings were characterized as amorphous Ca-P with high Ca/P stoichiometries with thicknesses of an order of magnitude difference (IBAD I = 30-50 nm and IBAD II = 300-500 nm). The histomorphometric and biomechanical testing results showed that the 300-500 nm thickness deposition (IBAD II) and PSHA positively modulated bone healing at early implantation times. PMID- 18508353 TI - Abrogation of the inflammatory response in LPS-stimulated RAW 264.7 murine macrophages by Zn- and Cu-doped bioactive sol-gel glasses. AB - The attenuation of an in vitro inflammatory response in RAW 264.7 murine macrophages stimulated with lipopolysaccharide (LPS) endotoxin was tested using sol-gel-derived bioactive glasses. Three general types of sol-gel-derived samples were evaluated: 58S, zinc-containing glasses, and copper-containing glasses. Distinct experimental procedures were used to test the potential of bioactive glasses to attenuate the inflammatory response in three situations: (1) therapeutically following LPS stimulation, (2) prophylactically before LPS stimulation of macrophages, and (3) indirectly via the glass dissolution products after stimulation with LPS. A sandwich enzyme-linked immunosorbent assay (ELISA) was used to monitor the concentration of tumor necrosis factor-alpha (TNF-alpha) secreted by macrophage cells. The strongest reduction in TNF-alpha concentration was observed when macrophage cells were first incubated with bioactive glass powder and then stimulated with LPS. This suggests a possible prophylactic application of these bioactive glasses for the prevention of inflammation. The 58S glass was capable of reducing the expression of TNF-alpha by macrophages, although the zinc- and copper-containing were more effective at suppressing the inflammatory response. The additional benefit of using zinc- and copper-doped bioactive glasses may be explained by the direct interactions of zinc and copper ions in key regulatory pathways for the inflammation response. PMID- 18508354 TI - The mechanism of deposition of calcium phosphate coatings from solution onto magnesium alloy AZ31. AB - In recent years, magnesium alloys have been proposed as a new class of metallic bioabsorbable implant material. Unfortunately, hydrogen gas evolution and an increase in alkalinity are both byproducts of the degradation process. This necessitates the development of magnesium alloys with controlled degradation rates. The development of biocompatible coatings that can delay the onset of corrosion is essential for improving the lifetime and performance of these materials in vivo. Calcium phosphate coatings have been shown to improve the biocompatibility of metallic implants for orthopedic applications. In this article, we report a solution chemistry technique for depositing calcium phosphate coatings on magnesium alloy surfaces. Our kinetic studies indicate that the deposition of the coating is related to the anodic dissolution of the substrate. Characterization of the coating by XPS, SEM/EDS, and XRD reveal that the coating produced is a poorly crystalline calcium magnesium hydroxyapatite material. PMID- 18508355 TI - Sinusoidal perfusion in the veno-occlusive region of living liver donors evaluated by indocyanine green and near-infrared spectroscopy. AB - Split liver transplantation and living donor liver transplantation (LDLT) commonly use a right liver graft without the middle hepatic vein (MHV). Although tributaries of the MHV are not reconstructed in the majority of cases, the alterations of the microcirculation and its regional functions remain unknown. We addressed these issues by assessing liver tissue indocyanine green (ICG) uptake with near-infrared spectroscopy (NIRS) in 21 donors. After graft procurement, visual inspection (before and after hepatic arterial clamping) and Doppler examination of the veno-occlusive region were performed. Bolus ICG (100 microg/kg) was then administered intravenously. Blood ICG at the finger tip was measured with pulse dye densitometry, whereas the liver ICG concentrations in the veno-occlusive and non-veno-occlusive regions were simultaneously measured for 15 minutes by NIRS. We estimated the hepatic ICG uptake rate constants in the veno occlusive region (Ku-oc) and non-veno-occlusive region (Ku-non). Changes in sinusoidal perfusion in the veno-occlusive region were expressed by the ratio of Ku-oc to Ku-non (Roc/non). The median value of Roc/non was 0.47, although it ranged from 0.13 to 0.94. Roc/non was related to the extent of liver surface discoloration before and after hepatic arterial clamping (P = 0.03 and 0.01, respectively). In conclusion, sinusoidal perfusion was impaired in the veno occlusive regions of living donor livers, but the magnitude of the effect varied greatly. Measurement of hepatic ICG uptake by NIRS could become a valuable tool for assessing the indication for venous reconstruction in LDLT and/or split donor liver transplantation. PMID- 18508356 TI - Ambient air pollution and risk for ischemic stroke and transient ischemic attack. AB - OBJECTIVE: Data on the association between air pollution and cerebrovascular disease in the United States are limited. The objective of this study was to investigate the association between short-term exposure to ambient air pollution and risk for ischemic cerebrovascular events in a US community. METHODS: Daily counts of ischemic strokes/transient ischemic attacks (TIAs) (2001-2005) were obtained from the population-based Brain Attack Surveillance in Corpus Christi (BASIC) Project. Daily particulate matter less than 2.5microm in diameter (PM(2.5)), ozone (O(3)), and meteorological data were obtained from Texas Commission on Environmental Quality. To examine the association between PM(2.5) and stroke/TIA risk, we used Poisson regression. Separate models included same day PM(2.5), PM(2.5) lagged 1 to 5 days, and an averaged lag effect. All models were adjusted for temperature, day of week, and temporal trends in stroke/TIA. The effects of O(3) were also investigated. RESULTS: Median PM(2.5) was 7.0microg/m(3) (interquartile range, 4.8-10.0microg/m(3)). There were borderline significant associations between same-day (relative risk [RR], 1.03; 95% confidence interval [CI], 0.99-1.07 for an interquartile range increase in PM(2.5)) and previous-day (RR, 1.03; 95% CI, 1.00-1.07) PM(2.5) and stroke/TIA risk. These associations were independent of O(3), which demonstrated similar associations with stroke/TIA risk (same-day RR, 1.02; 95% CI, 0.97-1.08; previous day RR, 1.04; 95% CI, 0.99-1.09). INTERPRETATION: We observed associations between recent PM(2.5) and O(3) exposure and ischemic stroke/TIA risk even in this community with relatively low pollutant levels. This study provides data on environmental exposures and stroke risk in the United States, and suggests future research on ambient air pollution and stroke is warranted. PMID- 18508357 TI - Liver transplantation for subacute hepatocellular failure due to massive steatohepatitis after bariatric surgery. AB - New therapeutic options for obesity include restrictive bowel surgery and surgery that promotes malabsorption, such as the Fobi-Capella (gastric bypass) and Scopinaro (biliopancreatic diversion) techniques. Complications associated with these procedures, such as hepatocellular failure, have been observed with increasing frequency. Reported here are 3 patients who, 7 to 24 months after bariatric surgery, developed hepatocellular failure, for which liver transplantation was considered to be indicated. Liver transplantation was undertaken in 2 of the patients; the third patient died while waiting for this procedure. We discuss the possible causes of this uncommon and poorly understood complication of surgery for obesity. One possibility is that it might arise as a result of progression of steatohepatitis. An alternative concept is that this complication may be secondary to rapid, massive loss of body weight. PMID- 18508358 TI - Effect of antibiotic prophylaxis on the risk of surgical site infection in orthotopic liver transplant. AB - Surgical site infections are common bacterial infections in orthotopic liver transplantation. The purpose of this study was to determine the incidence, timing, location, and risk factors, specifically antibiotic prophylaxis, for surgical site infections. A prospective study was performed that included a population of 1222 consecutive patients (73.0% males) who underwent liver transplantation in Spanish hospitals belonging to the Red de Estudio de la Infeccion en el Trasplante research network. One hundred seven patients developed surgical site infections. The predominant infection sites were incisional wound (53 episodes) and peritonitis (40 episodes). The timing of the organ/space surgical site infections was slightly delayed in comparison with incisional surgical site infections. Enterococcus spp., Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii were the predominant pathogens. Choledochojejunal or hepaticojejunal reconstruction (odds ratio, 4.2; 95% confidence interval, 1.6-10.7), previous liver or kidney transplant (odds ratio, 2.6; 95% confidence interval, 1.1-6.3), and more than 4 red blood cell units transfused (odds ratio, 2.0; 95% confidence interval, 1.1-3.4) were independently associated with the development of surgical site infections. Biliary reconstruction by choledochojejunostomy or hepaticojejunostomy increases the risk of surgical site infections. PMID- 18508359 TI - Glial fibrillary acidic protein as an early marker of hepatic stellate cell activation in chronic and posttransplant recurrent hepatitis C. AB - Activated alpha-smooth muscle actin (alpha-SMA)-positive hepatic stellate cells (HSCs) are pericytes responsible for fibrosis in chronic liver injury. The glial fibrillary acidic protein (GFAP), commonly expressed by astrocytes in the central nervous system, is expressed in vivo in the liver in a subpopulation of quiescent stellate cells. In the rat, increased GFAP expression in the acute response to injury and down-regulation in the chronic response have been observed, whereas reports concerning GFAP expression in human liver are still conflicting. We investigated the utility of GFAP compared to alpha-SMA as an immunohistochemical marker of early activated HSCs in chronic and posttransplant recurrent hepatitis C and correlated GFAP expression with vascular remodeling and fibrosis progression. With immunohistochemistry and a semiquantitative scoring system, the expression of GFAP and alpha-SMA in HSCs and the microvessel density were analyzed in biopsies from normal livers obtained from cadaveric donors [donor liver (DL); n = 21] and from livers from posttransplant hepatitis C virus recurrent hepatitis (HCV-PTR) patients (n = 19), hepatitis C virus chronic hepatitis (HCV-CH) patients, (n = 12), and hepatitis C virus cirrhosis (HCV-C) patients (n = 16). The percentage of alpha-SMA-positive HSCs was significantly higher in the HCV-PTR, HCV-CH, and HCV-C groups compared to the DL group (P < 0.01). The percentage of GFAP-positive HSCs was significantly higher in the HCV PTR group compared to the DL, HCV-C (P < 0.01), and HCV-CH (P < 0.05) groups and in the HCV-CH group compared to the DL group (P < 0.01), inversely correlating with the extent of fibrosis and microvessel density (P < 0.01). In the HCV-PTR group, the percentage of GFAP-positive HSCs correlated with fibrosis progression (P < 0.01). In conclusion, GFAP could represent a useful marker of early activation of HSCs in HCV-CH and seems to predict fibrosis progression in HCV PTR. PMID- 18508360 TI - Mouse toxicity of Anabaena flos-aquae from Lake Dianchi, China. AB - Some species of the genera Anabaena can produce various kinds of cyanotoxins, which may pose risks to environment and human health. Anabaena has frequently been observed in eutrophic freshwater of China in recent years, but its toxicity has been reported only in a few studies. In the present study, the toxicity of an Anabaena flos-aquae strain isolated from Lake Dianchi was investigated. Acute toxicity testing was performed by mouse bioassay using crude extracts from the lyophilized cultures. The mice exposed to crude extracts showed visible symptoms of toxicity and died within 10-24 h of the injection. Serum biochemical parameters were evaluated by the use of commercial diagnostic kits. Significant alterations were found in the serum biochemical parameters: alkaline phosphatase (AKP), gamma-glutamyl transpeptidase (gamma-GT), aspartate amino transferase (AST), alanine amino transferase (ALT), AST/ALT ratio, total protein content, albumin content, albumin/globulin (A/G) ratio, blood urea nitrogen (BUN), serum creatinine (Ssr), and total antioxidative capacity (T-AOC). Histopathological observations were carried out with hematoxylin and eosin (HE) stain under light microscope. Severe lesions were seen in the livers, kidneys, and lungs of the mice injected with crude extracts. The alterations of biochemical parameters were in a dose-dependent manner, and the severities of histological lesions were in the same manner. Based on biochemical and histological studies, this research firstly shows the presence of toxin-producing Anabaena species in Lake Dianchi and the toxic effects of its crude extracts on mammals. PMID- 18508361 TI - Oxidative stress, DNA damage, and antioxidant enzyme activity induced by hexavalent chromium in Sprague-Dawley rats. AB - Chromium is a widespread industrial compound. The soluble hexavalent chromium Cr (VI) is an environmental contaminant widely recognized as carcinogen, mutagen, and teratogen toward humans and animals. The fate of chromium in the environment is dependent on its oxidation state. The reduction of Cr (VI) to Cr (III) results in the formation of reactive intermediates leading to oxidative tissue damage and cellular injury. In the present investigation, Potassium dichromate was given intraperitoneally to Sprague-Dawley rats for 5 days with the doses of 2.5, 5.0, 7.5, and 10 mg/kg body weight per day. Oxidative stress including the level of reactive oxygen species (ROS), the extent of lipid peroxidation and the activity of antioxidant enzymes in both liver and kidney was determined. DNA damage in peripheral blood lymphocytes was determined by single-cell gel electrophoresis (comet assay). The results indicated that administration of Cr (VI) had caused a significant increase of ROS level in both liver and kidney after 5 days of exposure, accompanied with a dose-dependent increase in superoxide dismutase and catalase activities. The malondialdehyde content in liver and kidney was elevated as compared with the control animals. Dose- and time-dependent effects were observed on DNA damage after 24, 48, 72, and 96 h posttreatment. The results obtained from the present study showed that Cr (VI) could induce dose- and time dependent effects on DNA damage, both liver and kidney show defense against chromium-induced oxidative stress by enhancing their antioxidant enzyme activity. However, liver was found to exhibit more antioxidant defense than the kidney. PMID- 18508363 TI - Current diagnosis and management of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is an important liver disease with major morbidity and mortality. The diagnosis of PSC is confirmed by magnetic resonance cholangiopancreaticography, and endoscopic retrograde cholangiopancreaticography is performed in patients needing therapeutic endoscopy. As a result of the unknown cause of the disease, current medical therapies are unsatisfactory. Nevertheless, high-dose ursodeoxycholic acid should be recommended for treatment of PSC patients because there is a trend toward increased survival. Dominant bile duct stenoses should be treated endoscopically. However, liver transplantation continues to be the only therapeutic option for patients with advanced disease. Estimation of prognosis and timing of liver transplantation should be determined individually for each PSC patient on the basis of all results. The diagnosis and treatment of cholangiocarcinoma (CC) still remain a challenge in PSC patients. Early diagnosis of CC certainly is a prerequisite for successful treatment with surgical resection or innovative strategies such as neoadjuvant radiochemotherapy with subsequent orthotopic liver transplantation. Therefore, endoscopic techniques such as cholangioscopy and/or intraductal ultrasound may be useful diagnostic tools in patients with stenoses suspicious for malignancy. PMID- 18508364 TI - Foreword: ISTA 13 special issue. PMID- 18508365 TI - Regarding the risk for development of surgical site infections and bacterial prophylaxis in liver transplantation. PMID- 18508366 TI - Plasma cell hepatitis in liver allografts: Variant of rejection or autoimmune hepatitis? PMID- 18508367 TI - Identifying activated hepatic stellate cells in chronic and posttransplant recurrent hepatitis C. PMID- 18508368 TI - Biliary strictures following liver transplantation: past, present and preventive strategies. AB - Biliary complications are still the major source of morbidity for liver transplant recipients. The reported incidence of biliary strictures is 5%-15% after deceased donor liver transplantation and 28%-32% after right-lobe live donor surgery. Presentation is usually within the first year, but the incidence is known to increase with longer follow-up. The anastomotic variant is due to technical factors, whereas the nonanastomotic form is due to immunological and ischemic events, which later may lead to graft loss. Endoscopic management of anastomotic strictures achieves a success rate of 70%-100%; it drops to 50%-75% for nonanastomotic strictures with a higher recurrence rate. Results of endoscopic maneuvers are disappointing for biliary strictures after live donor liver transplantation, and the success rate is 60%-75% for anastomotic strictures and 25%-33% for the nonanastomotic variant. Preventive strategies in the cadaveric donor include the standardization of the type of anastomosis and maintenance of a vascularized ductal stump. In right-lobe live donor livers, donor liver duct harvesting also involves a major risk. The concept of high hilar intrahepatic Glissonian dissection, dissecting the artery and the duct as one unit, use of microsurgical techniques for smaller ducts, use of ductoplasty, and flexibility in the performance of double ductal anastomosis are the critical components of the preventive strategies in the recipient. In the case of live donors, judicious use of intraoperative cholangiograms, minimal dissection of the hilar plate, and perpendicular transection of the duct constitute the underlying principals for obtaining a vascularized duct. PMID- 18508369 TI - Prevention of hepatitis B recurrence after living donor liver transplantation: primary high-dose hepatitis B immunoglobulin monotherapy and rescue antiviral therapy. AB - The prevention of hepatitis B virus (HBV) recurrence is essential after liver transplantation in patients infected with HBV. We evaluated the efficacy of primary high-dose hepatitis B immunoglobulin (HBIG) monotherapy and rescue antiviral therapy in 639 HBV-infected adult patients who underwent living donor liver transplantation (LDLT) between February 1997 and December 2004. The overall 5-year survival rate was 80.7%, and recurrence of hepatocellular carcinoma was the most common cause of late mortality. Pretransplant HBV replication was observed in 392 (61.3%) patients. The interval of 10,000-IU HBIG administration to maintain antibody to hepatitis B surface antigen > 500 IU/L was 30 days in 11.4% patients, 40 to 50 days in 72.1%, and 60 days in 16.5%. At the last follow up, 3.9% of the patients without HBV recurrence were receiving combination therapy. Overall 1-year, 3-year, 5-year, and 10-year HBV recurrence rates were 1.4%, 5.5%, 7.3%, and 8.5%, respectively. HBV recurrence occurred after a mean of 25.7 +/- 16.4 months after LDLT. After HBV recurrence, 5 of 9 patients died from rapidly progressive liver failure before treatment with adefovir, and only 1 of 29 patients died after treatment with adefovir. Need for frequent HBIG infusions (< or =30 days), active pretransplant HBV replication, and hepatocellular carcinoma recurrence were significant risk factors for HBV recurrence and indications for combination therapy. Our posttransplant HBV prophylaxis regimen resulted in a 5-year HBV recurrence rate of 7.3% and a mortality rate of 13.2% after HBV recurrence, showing the effectiveness of high-dose HBIG monotherapy and rescue antiviral therapy. PMID- 18508370 TI - Hepatocellular carcinoma developed on compensated cirrhosis: resection as a selection tool for liver transplantation. AB - The objective of this study was to evaluate the histological profile obtained from primary resection of hepatocellular carcinoma (HCC) as a selection tool for liver transplantation (LT). The natural history of HCC depends on its histological features. The clinical effectiveness of resection as a selection tool for salvage or de principe LT has been previously advocated. Between 1987 and 2006, 20 patients underwent a resection prior to LT. Long-term survival of these 20 patients was compared to that of 73 patients who underwent primary LT. Histological features of the resected specimen were compared to those of the recurrences. Feasibility, morbidity, and mortality of LT following primary resection were also analyzed. Mean follow-up was 3.8 +/- 4.4 and 2.7 +/- 4.5 years from resection and LT, respectively; 6 patients died. The mean 1-, 3-, 5-, and 10-year overall survival rates were 71%, 61%, 55%, and 45% and 74%, 66%, 66%, and 40% after primary transplantation and primary resection, respectively (not significant). At LT, 14 patients had a recurrence, but histological study of the recurrence was not possible in 2 (complete necrosis). For 9 patients (75%), histological features of both primary and recurrent tumors were exactly the same. Four patients had recurrence following LT; in each case, primary and recurrent nodules shared the same histological markers of poor prognosis. De principe transplantation was proposed to 6 patients because of poor prognosis histological features on the resected specimen. All these patients are alive without recurrence with a mean follow-up of 55 months. In conclusion, the natural history of HCC can be predicted on the basis of the histological profile of the resected specimen, which may be used as a selection tool for LT. De principe LT in patients within Milan criteria with poor prognosis histological features may be an optimal strategy. PMID- 18508371 TI - Polysomnographic values in adolescents with ataxia telangiectasia. AB - Most adolescents with ataxia telangiectasia (A-T) develop progressive bulbar muscle weakness and decreased pulmonary reserve. The purpose of this study was to define the patterns of sleep and respiration during sleep, and to identify sleep related breathing problems in subjects with A-T. To address these issues, overnight polysomnography was performed on 12 adolescents with A-T. Eleven of the 12 subjects completed overnight polysomnography. The median age was 16 years (range, 13-20 years). All subjects in the study were wheelchair-bound and the median forced vital capacity (% predicted of normal) was 44% (range, 16-82%). The mean sleep efficiency was 72.6% with a mean apnea hypopnea index (AHI) of 0.7 events/hr (range, 0-2.2). The majority of apnea/hypopneas were REM related. The mean central apnea index was 0.1 events/hr (range, 0-0.2). The mean oxygen saturation nadir was 92.7% (range, 87-96) and the mean peak end-tidal carbon dioxide ET(CO(2) ) value was 53.8 mm Hg (range, 49-60). Two of 11 subjects had ET(CO(2) ) values >or=50 mm Hg for more than 50% of total sleep time. In this study, the majority of A-T adolescents had infrequent partial or complete upper airway obstructions during sleep and minimal nighttime hypoxemia. They did, however, have decreased sleep efficiency most likely, due in part, to their underlying neurological condition. This decrease in total sleep time may underestimate hypoventilation. Based on these findings, overnight polysomnography should be considered in adolescents with A-T, particularly in those in which there is a clinical suspicion of sleep related breathing abnormalities. PMID- 18508372 TI - Age, model for end-stage liver disease score, and organ functioning predict posttransplant tacrolimus neurotoxicity. AB - Calcineurin-inhibiting immunosuppressive medications are the mainstay of posttransplant immunosuppression. Although these highly beneficial drugs are critical for posttransplant survival, significant numbers of transplant recipients experience side effects, some requiring a switch to a different immunosuppressive regimen. Neurotoxicity is one of the most debilitating side effects because of its impact on mental status and cognition. As our center uses tacrolimus as the initial immunosuppressant for all liver transplant (LTX) recipients, we were interested in those patients who required a switch because of neurotoxic side effects. Over a 5-year period, 827 adult LTX recipients received their first graft at our center. Ninety-four patients were no longer on tacrolimus by 2 months post-LTX (86 switched because of concerns over neurotoxicity, and 8 switched because of renal function concerns). Of those experiencing neurotoxic side effects, the majority (64%) had altered mental status, and 26% had seizures (first onset post-LTX). On the basis of our prior work, we hypothesized that patients with a pre-LTX history of excessive alcohol use would be at higher risk for neurotoxic effects. We also hypothesized that the elderly and those who had more advanced illness (that is, higher Model for End Stage Liver Disease scores) at LTX would be at risk as well. We found that patients with a pre-LTX diagnosis of alcoholic liver disease were not more likely to be switched from tacrolimus. Furthermore, we found that in addition to older age and higher Model for End-Stage Liver Disease scores, poorer hepatic functioning was significantly associated with a switch from tacrolimus. We discuss the implications of these findings and the relevance for future clinical care in these high-risk patients. PMID- 18508373 TI - Preoperative dobutamine stress echocardiographic findings and subsequent short term adverse cardiac events after orthotopic liver transplantation. AB - Cardiovascular (CV) complications are the leading cause of non-graft-related death in orthotopic liver transplant (OLT) patients. Pretransplant cardiac evaluation using dobutamine stress echocardiography (DSE) is commonly utilized for risk stratification of OLT candidates. To determine if clinical and echocardiographic variables identify patients with increased CV risk, we performed a retrospective chart review of all 284 patients that underwent OLT at our institution between June 1999 and August 2005. Of these patients, 157 had a DSE prior to their OLT. Serious adverse CV events occurring during surgery and up to 4 months post-transplantation were defined as cardiac-related death, myocardial infarction (MI), new heart failure, or asystole or unstable ventricular arrhythmia requiring acute treatment. Sixteen of 157 patients (10%) had an adverse CV event with 2 deaths. These included ventricular tachycardia (n = 8), asystole (n = 2), MI (n = 5), and new heart failure (n = 1). Nine of the 16 CV events occurred at the time of surgery (including both deaths), 5 occurred postoperatively, and 3 occurred after hospital discharge. Variables that correlated with increased CV events were inability during DSE to achieve >82% of the maximum predicted heart rate (22% versus 6%, P = 0.01), a peak rate pressure product during DSE of <16,333 (17% versus 5%, P = 0.02), and a Model for End Stage Liver Disease (MELD) score of >24 at the time of OLT. A multivariate model calculated from the DSE maximum achieved heart rate (MAHR) and MELD score (result = 3.78 + 0.07 MELD - 0.05 MAHR) identified a 47% risk for a value > 0 versus a 6% risk for a value < 0 (P < 0.001). In conclusion, the maximum heart rate achieved during DSE together with the MELD score may be a predictor of adverse CV events up to 4 months post-OLT. A large prospective study is needed to more decisively support this conclusion. PMID- 18508374 TI - ImmuKnow: a new parameter in immune monitoring of pediatric liver transplantation recipients. AB - Lifelong immunosuppression is mandatory for optimal graft and patient survival following liver transplantation. Nevertheless, graft rejection or numerous adverse events associated with overimmunosuppression or underimmunosuppression cannot be completely avoided. The ImmuKnow assay measures cell-mediated immunity and is able to discern between conditions of overimmunosuppression and underimmunosuppression. The aim of this study was to evaluate the ImmuKnow assay in the evaluation of the immune function in pediatric liver transplant recipients and to assess its correlation with the patients' clinical and biochemical status. Eighty-nine whole blood samples were collected from 23 liver transplant recipients that were 1 to 18 years old. The net state of immune function was determined by the quantitative measurement of the intracellular adenosine 5 triphosphate level in CD4+ lymphocytes after phytohemagglutinin stimulation. Comprehensive clinical data were correlated with the ImmuKnow assay results. In 23 of the 28 samples collected during clinical quiescence, ImmuKnow results were correlated with the clinical status, expressing the patient's moderate immune function. However, a correlation between measured therapeutic drug levels and clinical quiescence was found in only 18 of the 28 samples. In 6 patients who suffered from clinical complications, ImmuKnow measurements showed a wide range of deviations, expressing the unstable immunological status of these patients. In conclusion, the ImmuKnow assay correlates with the clinical status of liver transplanted children. It serves as a reliable and unique parameter of the cellular immune function. We conclude that the ImmuKnow assay, together with existing clinical tools, may allow for the immune monitoring of pediatric liver recipients. PMID- 18508375 TI - When is a liver transplant futile? PMID- 18508376 TI - Role of Kupffer cells in the induction of tolerance of orthotopic liver transplantation in rats. AB - Because the role of Kupffer cells (KCs) in liver transplantation (LT) tolerance is not well understood, we investigated their role in liver allograft acceptance in rats. Male Sprague-Dawley rats were randomly assigned to either an LT group or a transplantation group pretreated with GdCl(3) (Gd group). The rats were postoperatively sacrificed at indicated times for histology and assessment of KC function, nuclear factor kappa B (NF-kappaB) activity, and cytokine production. KCs and T cells (TCs) were isolated from allografts to assess Fas/Fas ligand (FasL) expression. Cytotoxicity of KCs against TCs was monitored by coculturing of (3)H-thymidine TCs with KCs at various effector-to-target ratios. The results were as follows. First, grafts were spontaneously accepted in the LT group with evident apoptosis of TCs; however, inhibition of KCs by pretreatment with GdCl(3) decreased TC apoptosis and shortened the survival of allografts. Second, KCs in the LT group had increased levels of FasL messenger RNA and protein with respect to that in the Gd group. Third, by in vitro cocultivation assays, KCs induced TC apoptosis though elevated expression of FasL, and this process could be blocked by anti-FasL antibody. Fourth, there was a positive correlation between activation of NF-kappaB and FasL expression in KCs and interleukin-4 production in the LT group, and the activation of NF-kappaB was inhibited by pretreatment with GdCl(3). In conclusion, KC-induced depletion of TCs via the Fas/FasL pathway might play a critical role in LT tolerance. However, the tolerance is abrogated by suppression of FasL and IL-4 expression via inhibition of NF-kappaB activity by GdCl(3). PMID- 18508377 TI - Comparison of four model for end-stage liver disease-based prognostic systems for cirrhosis. AB - Serum sodium (Na) has been suggested for incorporation into the Model for End Stage Liver Disease (MELD) to enhance its prognostic ability for patients with cirrhosis. Three Na-containing models--the Model for End-Stage Liver Disease with the incorporation of serum sodium (MELD-Na), the integrated Model for End-Stage Liver Disease (iMELD), and the Model for End-Stage Liver Disease to sodium (MESO) index--were independently proposed for this purpose. This study investigated the accuracy of these 4 MELD-based models for outcome prediction. The c-statistic equivalent to the area under the receiver operating characteristic curve (AUC), used to predict 3- and 6-month mortality, was calculated and compared in 825 patients with cirrhosis. The MELD score tended to be lower with increasing Na level. At 3 months of enrollment, the iMELD had the highest AUC (0.807) and was followed by the MELD-Na (0.801), MESO (0.784), and MELD (0.773); the difference between the MESO and MELD was statistically significant (P = 0.013). At 6 months, the iMELD still had the highest AUC (0.797) and was followed by the MELD-Na (0.778), MESO (0.747), and MELD (0.735); all comparisons showed significant differences between each other (all P < 0.01), with the exception of iMELD and MELD-Na (P = 0.18). With the most discriminative cutoffs, the specificity and negative predictive value were 70%-85% and 89%-97%, respectively, at 3 and 6 months for the 4 models. Patients with spontaneous bacterial peritonitis (SBP) consistently had significantly higher MELD-derived scores in all 4 models compared to patients without SBP (all P < 0.01). Patients with hepatic encephalopathy also had higher scores in all 4 models, although the statistical significance was established only for the iMELD (41.0 +/- 11.5 versus 37.6 +/- 9.1, P = 0.037). In conclusion, the incorporation of Na into the MELD may enhance prognostic accuracy. Both the iMELD and MELD-Na are better prognostic models for outcome prediction in patients with cirrhosis. Patients with SBP have a higher MELD-derived score. Future studies are warranted to define the optimal MELD-based prognostic model for cirrhosis. PMID- 18508378 TI - The easiest way to improve the outcome of suboptimal liver grafts. PMID- 18508379 TI - The prevalence of a heparin-like effect shown on the thromboelastograph in patients undergoing liver transplantation. AB - It has been known for several decades that thromboelastographic analysis of the blood of patients undergoing liver transplantation may show a heparin-like effect (HLE) at the time of reperfusion. However, the prevalence of HLE and the origin of these heparin-like substances remain largely unstudied. The primary aim of this retrospective observational analysis was to determine the prevalence of the HLE in 211 consecutive patients having liver transplantation in our institution at various stages throughout the transplant. One of the secondary aims was to analyze the prevalence of HLE with respect to the various etiologies of liver disease. Paired Thromboelastograph traces (native and heparinase) were examined at 5 stages of the transplant: the baseline stage, dissection stage, anhepatic stage, reperfusion stage, and end of the case. HLE was defined as a reduction in the reaction and coagulation times of greater than 50% by the addition of heparinase to the sample. Thirty-one percent of patients had evidence of an HLE at baseline, and this increased to 75% after reperfusion of the donor graft. This HLE resolved spontaneously in 47% by the end of the case. Patients with fulminant liver failure were more likely to demonstrate HLE at baseline than those with chronic liver disease (45.8% compared to 29%). There was no difference in the prevalence of HLE after reperfusion. In conclusion, prior to transplantation, there is a significant difference in the prevalence of HLE with respect to etiology. However, this difference disappears after reperfusion as the majority of patients then develop HLE. Although it is clear that there are both endogenous and exogenous sources of heparin contributing to the HLE, the clinical significance of these findings remains unclear. PMID- 18508382 TI - Posttransplant plasma cell hepatitis (de novo autoimmune hepatitis) is a variant of rejection and may lead to a negative outcome in patients with hepatitis C virus. AB - De novo autoimmune hepatitis has been described in both pediatric and adult liver transplantation (LT) recipients. Studies of small numbers of patients have proposed it to be an alloimmune hepatitis or form of chronic rejection. We have recently noted an increasing number of patients with post-LT recurrent hepatitis C virus (HCV) developing this, with an apparent negative impact on outcome and survival. We term this entity posttransplant plasma cell hepatitis (PCH). A search of our institution's pathology database was performed with the terms "plasma cell(s)," "lymphoplasmacytic infiltrate," and "liver allograft." A histological scoring system was devised to more reliably diagnose PCH in the setting of recurrent HCV. Thirty-eight patients were identified, and their clinical data were analyzed. Sixty percent had a negative outcome as defined by the development of cirrhosis, need for retransplantation, or death. Eighty-two percent had recent lowering of immunosuppression or subtherapeutic calcineurin inhibitor levels; 58% developed PCH within 2 years post-LT. Histologic resolution of PCH was associated with good outcome (P < 0.001). Patients not receiving treatment had a negative outcome (P = 0.007) as did patients receiving corticosteroids as therapy (P = 0.02). Persistence (P = 0.007) or recurrence of PCH was associated with negative outcome. In conclusion, PCH is a histologic variant of rejection. On liver biopsy, PCH can at times be difficult to diagnose, and the use of a standardized scoring system is recommended to differentiate it from other forms of allograft dysfunction. Treatment by optimization of immunosuppression without the use of corticosteroids appears effective. The development of PCH in the setting of recurrent HCV is a negative prognostic factor for patient outcome and allograft failure. PMID- 18508383 TI - Effects of the nematode Phasmarhabditis hermaphrodita and of venom from the endoparasitic wasp Pimpla hypochondriaca on survival and food consumption of the pest slug Deroceras reticulatum; implications for novel biocontrol strategies. AB - BACKGROUND: Controlling pests through disruption of biochemical pathways by physiologically active compounds/factors from animals and plants represents an expanding field of research. The authors investigated whether such factors in venom from the wasp Pimpla hypochondriaca (Retzius) can affect the viability and food consumption of the slug Deroceras reticulatum (Muller), and whether they can improve the efficacy of nematode-induced slug mortality. RESULTS: Exposure of slugs to 4 mL of water containing 500, 1000 and 5000 Phasmarhabditis hermaphrodita (Schneider) resulted in significant increases in mortality (with hazard ratios of 3.5, 3.9 and 5.8 respectively) and significant reductions in total food consumption and mean food consumption each day for 21 days. Injection of slugs with 4, 8 or 12 microL of P. hypochondriaca venom resulted in significant increases in mortality (with hazard ratios of 3.3, 4.5 and 9.0 respectively) and significant reductions in total food consumption compared with the controls. However, there was no significant effect of venom on the mean food consumption on individual days of the 21 day assay period, although significant reductions occurred for the 8 and 12 microL doses up to day 10. Injecting slugs with 4 microL of venom prior to exposure to 500 nematodes had no synergistic effect on either mortality or food consumption compared with either of the individual treatments. CONCLUSIONS: Pimpla hypochondriaca venom contains factors capable of killing and reducing food consumption by D. reticulatum. The utilization of these factors as components of integrated pest management strategies is discussed. PMID- 18508384 TI - Nitric oxide-mediated activation of NADPH oxidase by salbutamol during acute asthma in children. AB - A wide variety of reactive oxygen species (ROS) such as superoxide anion, hydroxyl radical and hydrogen peroxide, and reactive nitrogen species such as nitric oxide (NO) and peroxynitrite are known to be involved in pathophysiology of bronchial asthma. We have investigated, in this study, the status of NADPH oxidase (NOX), a major source of superoxide anion production, in peripheral blood lymphocytes (PBL) from asthmatic patients in relation to salbutamol treatment. PBL isolated from patients with bronchial asthma were found to have a significantly increased activity of NOX. Plasma levels of malondialdehyde (MDA), an index of lipid peroxidation, and NO were also markedly elevated in asthmatic patients compared to control samples. A significantly decreased catalase activity observed in PBL from our patients underscored the severity of oxidative stress during asthma. Treatment of PBL with salbutamol (10 microg ml(-1)), prevented the attenuation of catalase activity but significantly increased the levels of NO and NOX activity. Levels of NOX-1 mRNA were significantly (p < 0.001) increased in PBL following treatment with NO donor (500 microM), S-nitroso-N-acetyl penicillamine (SNAP). Western blot analysis revealed that gp91phox protein was also significantly (twofold-threefold) increased following treatment with SNAP. The observed transcriptional regulation of NOX-1 and gp91phox by NO was observed to result in an increased NOX activity as well. This study concludes that salbutamol treatment enhances superoxide anion production in asthma patients through NO-mediated mechanisms, however it exerts beneficial antioxidant effects through activation of catalase and attenuation of lipid peroxidation. PMID- 18508385 TI - In vitro effects of 2-methoxyestradiol on MCF-12A and MCF-7 cell growth, morphology and mitotic spindle formation. AB - The influence of 2-methoxyestradiol (2ME) was investigated on cell growth, morphology and spindle formation in a tumorigenic (MCF-7) and non-tumorigenic (MCF-12A) epithelial breast cell line. Inhibition of cell growth was more pronounced in the MCF-7 cells compared to the MCF-12A cells following 2ME treatment. Dose-dependent studies (10(-5)-10(-9) M) revealed that 10(-6) M 2ME inhibited cell growth by 44% in MCF-12A cells and by 84% in MCF-7 cells (p-value < 0.05). 2ME-treated MCF-7 cells showed abnormal metaphase cells, membrane blebbing, apoptotic cells and disrupted spindle formation. These observations were either absent or less prominent in MCF-12A cells. 2ME had no effect on the length of the cell cycle between S-phase and the time a mitotic peak was reached in either cell line but MCF-7 cells were blocked in mitosis with no statistically significant alterations in the phosphorylation status of Cdc25C. Nevertheless, Cdc2 activity was significantly increased in MCF-7 cells compared to MCF-12A cells (p-value < 0.05). The results indicate that 2ME disrupts mitotic spindle formation and enhances Cdc2 kinase activity, leading to persistence of the spindle checkpoint and thus prolonged metaphase arrest that may result in the induction of apoptosis. The tumorigenic MCF-7 cells were especially sensitive to 2ME treatment compared to the normal MCF-12A cells. Therefore, differential mechanism(s) of growth inhibition are evident between the normal and tumorigenic cells. PMID- 18508386 TI - Effect of concanavalin A (Con-A) on the hormone production of the unicellular Tetrahymena and the immune cells of the rat. A comparative study. AB - Tetrahymena populations were treated with 10(-15) g ml(-1) or 10(-6) g ml(-1) concanavalin-A (Con-A) in tryptone-yeast medium for 1 h. Rat peritoneal immune cells (mast cells, lymphocytes, monocyte-granulocyte group) were also treated with 10(-6) g ml(-1) Con-A, for 1 h. The cells' hormone (ACTH, histamine, serotonin, endorphin, triiodothyronine (T(3))) content was measured by using immunocytochemistry and flow cytometry. The extremely low dose of Con-A universally and significantly elevated the hormone contents, while the result of higher dose was uncertain. In the immune cells, Con-A significantly decreased the ACTH level in each cell type and histamine level in mast cells. The results demonstrate the very high sensitivity of Tetrahymena receptors for a non-hormone (lectin) molecule, which can bind to the insulin receptors and mimics the effect of insulin. The results also show that Tetrahymena receptors are more sensitive to lower concentrations of molecules than to higher ones. The universal hormone production stimulating effect of Con-A-which is observed in Tetrahymena-is specified in rat. PMID- 18508387 TI - Effects of sodium fluoride (NaF) on the cilia and microtubular system of Tetrahymena. AB - The effect of the nucleophilic reagent NaF on the microtubular system of Tetrahymena was studied by using scanning electron microscopy (SEM), confocal microscopy, and flow cytometry. Treatments with 40 mM NaF significantly reduced the amount of alpha-tubulin while 80 mM treatment did not alter its quantity. One possible explanation for this alpha-tubulin overexpression is that the higher amount of alpha-tubulin enables this organism to carry out the appropriate function of the cytoskeleton under this undesirable influence of higher amounts of 80 nM NaF. However, the amount of acetylated tubulin increased in a dose dependent manner. The cilia became fragile under the effect of 80 mM NaF. Confocal microscopy revealed that after 40 mM NaF treatment transversal microtubule bands (TMs) and longitudinal microtubule bands (LMs) as well as basal bodies (BBs) were extremely strong decorated with anti-acetylated tubulin antibody and TM-localization abnormalities were visible. In the 80 mM NaF-treated cells, the deep fiber of oral apparatus was very strongly labeled, while the TMs and LMs were less decorated with anti-acetylated tubulin antibody, and LM deformities were visible. It is supposed that post-translational tubulin modifications (e.g., acetylation) defend the microtubules against the NaF-induced injury. NaF is able to influence the activity of several enzymes and G-proteins, therefore is capable to alter the structure, metabolism, and the dynamics of microtubular system. The possible connection of signaling and cytoskeletal system in Tetrahymena is discussed. PMID- 18508388 TI - Mitochondrial AIF protein involved in skeletal muscle regeneration. AB - The mitochondrial flavoprotein apoptosis-inducing factor (AIF) has proved to be either the main mediator of apoptosis or an anti-apoptotic factor via its putative oxidoreductase and peroxide scavenging activities. We report here that 100 muM hydrogen peroxide (H2O2) induced the proliferation of C2C12 myoblasts and over-expression of AIF simultaneously in vitro. Immunofluorescence showed that the over-expression of AIF was located in the cytoplasm. The immunopositive AIF was detected in nuclei 27 days after denervation of skeletal muscle, but in the cytoplasm it was detected 27 days after fiber-damaged skeletal muscle. AIF may be a factor involved in skeletal muscle regeneration. PMID- 18508389 TI - 4-Amino-3-acetylquinoline-induced apoptosis of murine L1210 leukemia cells involves ROS-mitochondrial-mediated death signaling and activation of p38 MAPK. AB - Quinolines are known to be multitarget agents with a broad spectrum of biological activity. In a previous study, we showed that newly prepared 4-amino-3 acetylquinoline (AAQ) possesses strong anticancer activities. In this study, we investigated whether AAQ has cytotoxicity in murine L1210 leukemia cells. Results from cell proliferation assays showed that AAQ caused significant decrease in cell number in a dose-dependent manner. The cell death induced by AAQ appeared to involve apoptosis, based on evidence from apoptotic DNA fragmentation, flow cytometry, fluorescence microscopy, and Western blot analyses. We found that AAQ treated cells had activated p38 MAPK and that apoptosis was processed through a reactive oxygen species (ROS)-dependent mitochondrial pathway. In summary, our results suggest that AAQ can induce apoptosis, at least in part, through the activation of the p38 MAPK pathway in L1210 leukemia cells. PMID- 18508390 TI - Mitochondria as the target for mildronate's protective effects in azidothymidine (AZT)-induced toxicity of isolated rat liver mitochondria. AB - Previously mildronate, an aza-butyrobetaine derivative, was shown to be a cytoprotective drug, through its mechanism of action of inhibition of carnitine palmitoyltransferase-1, thus protecting mitochondria from long-chain fatty acid accumulation and subsequent damage. Recently in an azidothymidine (AZT)-induced cardiotoxicity model in vivo (in mice), we have found mildronate's ability of protecting heart tissue from nuclear factor kappaB abnormal expression. Preliminary data also demonstrate cerebro- and hepatoprotecting properties of mildronate in AZT-toxicity models. We suggest that mildronate may target its action predominantly to mitochondria. The present study in isolated rat liver mitochondria was designed to clarify mitochondrial targets for mildronate by using AZT as a model compound. The aim of this study was to investigate: (1) whether mildronate may protect mitochondria from AZT-induced toxicity; and (2) which is the most critical target in mitochondrial processes that is responsible for mildronate's regulatory action. The results showed that mildronate protected mitochondria from AZT-induced damage predominantly at the level of complex I, mainly by reducing hydrogen peroxide generation. Significant protection of AZT caused inhibition of uncoupled respiration, ADP to oxygen ratio, and transmembrane potential were also observed. Mildronate per se had no effect on the bioenergetics, oxidative stress, or permeability transition of rat liver mitochondria. Since mitochondrial complex I is the first enzyme of the respiratory electron transport chain and its damage is considered to be responsible for different mitochondrial diseases, we may account for mildronate's effectiveness in the prevention of pathologies associated with mitochondrial dysfunctions. PMID- 18508392 TI - Impact of hip fracture, heart failure and weight loss on the risk of institutionalization of community-dwelling patients with dementia. AB - OBJECTIVES: This study sought to identify the influence of medical symptoms and diseases on the risk of nursing home placement in a prospective cohort of newly diagnosed community-dwelling patients with dementia. STUDY DESIGN AND SETTING: This study included 348 patients with dementia, consecutively diagnosed, recruited and followed at a geriatric outpatient center (mean age: 81 years, 65.5% with Alzheimer's disease, mean baseline MMSE score: 20.5, mean follow-up: 20.5 months). RESULTS: After adjustment for factors commonly associated with institutionalization in this population, hip fracture in the 3 years preceding diagnosis, acute congestive heart failure during follow-up and weight loss of more than 5% in any year during follow-up were independently associated with nursing home placement. CONCLUSION: This study confirms the independent contribution of specific medical symptoms and diseases to earlier institutionalization of patients with dementia. These results stress the importance of better knowledge of the specific characteristics of hip fracture, weight loss and congestive heart failure in the context of dementia, to make more effective prevention possible in this patient population. PMID- 18508393 TI - Prevalence of cholinesterase inhibitors in subjects with dementia in Europe. AB - PURPOSE: To evaluate the prevalence of cholinesterase inhibitor (ChI) treatment in subjects with dementia in European countries. METHODS: We studied the prevalence of treatment in subjects with dementia among European countries in 2004 (Belgium, France, Germany, Italy, the Netherlands, Poland, Portugal, Spain and the United Kingdom) by using estimates of prevalence of dementia and of ChI treatments according to sales and reimbursement data. RESULTS: In 2004, estimated prevalence of ChI use among subjects with dementia ranged from 3.0% in the Netherlands to 20.3% in France. It was 17.5% in Spain, 6.7% in the UK and 5.9% in Italy. Donepezil was used by more than 60% of patients using a single ChI and represented almost 50% of reimbursements for patients that had used at least two different ChIs during the year. Galantamine and rivastigmine were respectively used by 22 and 18% of subjects using a single drug and 27 and 23% of reimbursements for patients that had used at least two different ChIs. Nevertheless, different patterns of use were found for individual countries. CONCLUSIONS: Prevalence of treatment by ChIs among subjects with dementia remains weak and varies greatly across Europe. Differences in reimbursement rates and health policies could partly explain these variations, as ChIs could have failed to convince health authorities because the outcomes considered for trials are not used by clinicians in their everyday practice. If donepezil was highly predominant across countries, variations in rivastigmine and galantamine importance could reflect local market specificities. PMID- 18508394 TI - Involvement of hippocampal nitric oxide in spatial learning in the rat. AB - Nitric oxide (NO) is thought to be involved in synaptic plasticity contributing to learning and memory in several brain areas including the hippocampus. The hippocampus is believed to have a critical role in the processing of spatial information. But, data on the role of hippocampal NO in spatial learning are not consistent. So the effect of NO synthase (NOS) inhibition in the CA1 region of rat hippocampus on spatial localization was investigated in the Morris water maze (MWM). Male albino Wistar rats cannulated in their CA1 region received bilateral injections of vehicle (saline) or N(omega)-nitro-L-arginine methyl ester (L NAME), a NOS inhibitor (50, 100 and 200 microg/0.5 microl) through the cannulae 30 min before training each day. Animals were subjected to 5 days of training in the MWM; 4 days with the invisible platform to test spatial learning and the 5th day with the visible platform to test motivation and sensorimotor coordination. The results showed dose-dependent increases (p<0.001) in escape latency, traveled distance, heading angle, and dose-dependent decreases (p<0.01) in target quadrant entries in L-NAME-received groups as compared to the control group. This impairment was reversed by co-administration of mole-equivalent doses of L arginine (L-Arg), the NO precursor. L-Arg alone at the dose of 129.2 microg, increased heading angle (p<0.01) with no effect on other parameters. On the basis of the present data, it is concluded that processes mediated by NO synthesis in the hippocampus are essentially involved in spatial learning. PMID- 18508395 TI - Potential of enterococci isolated from horses. AB - Faecal samples of 122 horses (from farms in Slovakia) were examined to select enterococci to study their probiotic potential for their further use as additives. Each gram of faeces contained 1.0-5.0 cfu (log 10) of enterococci. Of the 43 isolates, 25 (58.1%) were identified as Enterococcus faecium, 3 strains were (6.9%) Enterococcus mundtii and one strain was identified as E. faecalis. Fourteen isolates were not characterized further. A significant proportion of the isolates were resistant to kanamycin, vancomycin and gentamicin. Low urease activity of enterococci dominated. The values of lactic acid ranged from 0.98 to 1.91 mmol/L. Porcine fibronectectin and bovine lactoferrin were bound weakly by tested enterococci, while bovine fibrinogen was bound more strongly. Enterococci from horses did not bind bovine apotransferrin. The isolates adhered with the same ability to human as well as to canine mucus. At least one enterocin gene was detected among 16 analyzed isolates. Ent B gene was detected in all strains tested (16, 100%), followed by the genes ent A, ent P and ent L50B. Three suitable candidates-the strains of E. faecium EF 412, EF 462 and EF 491 were selected for further detail studies and possibilities to be used as additives. PMID- 18508396 TI - Analysis of the proteins involved in the structure and synthesis of the cell wall of Ustilago maydis. AB - A study of the proteins involved in the synthesis and structure of the cell wall of Ustilago maydis was made by in silico analysis of the fungal genome, with reference to supporting experimental evidence. The composition of the cell wall of U. maydis shows similarities with the structural composition of the walls of Ascomycetes, but also shows important differential features. Accordingly, the enzymes involved in the synthesis of the U. maydis wall polysaccharides chitin and beta-1,6 glucans displayed some differential characteristics. The most salient difference in protein composition was the predicted absence of Pir proteins, an important class of proteins present in the Ascomycetes. Other classes of proteins that are covalently-linked to the wall in Ascomycetes, including those bound through disulfide linkages, joined by alkali-labile bonds, and GPI proteins, were predicted to be present in the U. maydis walls. The main characteristic of the exo-cellular, non-covalently-bound proteins was their relative low number, especially for hydrolytic enzymes. PMID- 18508397 TI - Rearing temperature enhances hepatic glucokinase but not glucose-6-phosphatase activities in European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) juveniles fed with the same level of glucose. AB - The aim of this work was to elucidate if the previous results observed in hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities in European sea bass and gilthead sea bream are due to temperature per se or to differences in feed intake at different water temperatures. For that purpose triplicate groups of fish (30 g initial body weight) were kept at 18 degrees C or 25 degrees C during two weeks and fed a fixed daily ration of a glucose-free or 20% glucose diet. At the end of the experimental period, plasma glucose levels in both species were not influenced by water temperature but were higher in fish fed the glucose diet. Higher hepatic GK activity was observed in the two fish species fed the glucose diet than the glucose-free diet. In the glucose fed groups, GK activity was higher at 25 degrees C than at 18 degrees C. Glucose-6-phosphatase activities in both species were not influenced by water temperature. In European sea bass and in contrast to gilthead sea bream it was observed an effect of dietary composition on G6Pase activities with surprising higher activities recorded in fish fed the glucose diet than in fish fed the glucose-free diet. Overall, our data strongly suggest that European sea bass and gilthead sea bream are apparently capable to strongly regulate glucose uptake by the liver but not glucose synthesis, which is even enhanced by dietary glucose in European sea bass. Within limits, increasing water temperature enhances liver GK but not G6Pase activities, suggesting that both species are more able to use dietary carbohydrates at higher rearing temperatures. PMID- 18508398 TI - An allometric study of fatty acids and sensitivity to lipid peroxidation of brain microsomes and mitochondria isolated from different bird species. AB - The objective of this investigation was to examine the relationship between body size, fatty acid composition and sensitivity to lipid peroxidation of mitochondria and microsomes isolated from the brain of different size bird species: manon, quail, pigeon, duck and goose, representing a 372-fold range of body mass. Fatty acids of total lipids were determined using gas chromatography and lipid peroxidation was evaluated using a chemiluminescence assay. The allometric study of the fatty acids present in brain mitochondria and microsomes of the different bird species showed a small number of significant allometric trends. In mitochondria the percentage of monounsaturated fatty acids, was significantly lower in the larger birds (r=-0.965; P<0.008). The significant allometric increase in 18:2 n-6; linoleic acid (r=0.986; P<0.0143), polyunsaturated (r=0.993; P<0.007) and total unsaturated (r=0.966; P<0.034) in brain microsomes but not in mitochondria may indicate a preferential incorporation of this fatty acid in the brain endoplasmic reticulum of the larger bird species. The brain of all birds studied had a high content of docosahexaenoic acid. However brain mitochondria but not microsomes isolated from all the birds analyzed showed a significant decrease of arachidonic and docosahexaenoic acids during lipid peroxidation. The allometric analyses of chemiluminescence were not statistically significant. In conclusion our results show absence of correlation between the sensitivity to lipid peroxidation of brain mitochondria and microsomes with body size and maximum life span. PMID- 18508399 TI - Parvalbumin characterization from the euryhaline stingray Dasyatis sabina. AB - The Atlantic stingray, Dasyatis sabina found along the Gulf of Mexico and southeastern Atlantic coasts, is a euryhaline species of elasmobranch. This species is able to osmotically compensate for changing environmental salinity by altering plasma and intracellular solutes, including urea and counteracting methylamines (betaine and TMAO). Parvalbumin (PV) is an intracellular protein that facilitates muscle relaxation by sequestering calcium. Determining the effects that in situ concentrations of urea (146 mM), betaine (62 mM), and TMAO (11 mM) have on PV function in marine and freshwater adapted populations of D. sabina could provide insight into intracellular correlates of euryhaline tolerance for this species. PV from marine and freshwater populations of D. sabina was identified and purified by SDS-PAGE, western blot analysis, and full amino acid sequence analysis. Both populations exhibited two PV isoforms, PV I (approximately 12.18 kDa mw) and PV II (11.96 kDa mw). PV dissociation constants (K(D)) were determined in the presence and absence of physiological concentrations of urea, betaine, and TMAO by fluorescence spectroscopy using the fluorescent Ca(2+) indicator fluo-3 which competes with PV for Ca(2+). Functional studies revealed PV I showed no significant changes in calcium binding from in situ muscle conditions, except in the presence of betaine. In contrast, PV II's ability to bind calcium was increased relative to physiological conditions in the presence of each osmolyte independently. Thus, it appears that organic osmolytes have isoform specific effects on PV function. PMID- 18508400 TI - An analysis of IGFBP evolution. AB - OBJECTIVES: To perform a synonymous, non-synonymous codon mutational analysis of the IGFBP gene family and identify mechanisms by which the IGFBP subfamilies diverged. METHODS: We identified 78 intact nucleotide sequences from 6 IGFBP subfamilies and 12 different species and used them for phylogenetic and synonymous, non-synonymous codon mutational analysis. Deletion and insertion comparisons were performed across subfamilies to determine if this might play a unique role in subfamily genesis. RESULTS: IGFBP-2 was identified by phylogenetic analysis to be the most related subfamily of the IGFBP progenitor, followed by IGFBP-4. Insertions and deletions within the variable domains were associated with divergence of each subfamily from its progenitor, suggesting a common motif for IGFBP evolution. Insertions unique to mammals were also found within the amino terminus of IGFBP-2. CONCLUSION: IGFBP subfamily divergence is associated with variable domain insertion or deletion and vigorous non-synonymous codon mutation. Our findings suggest strong selective pressure for IGFBP divergence in terrestrial vertebrates. PMID- 18508401 TI - Fungal malignant otitis externa treated with hyperbaric oxygen. AB - OBJECTIVE: To report a case of Aspergillus flavus malignant otitis externa, successfully treated with antifungal agents, surgical debridement, and hyperbaric oxygen treatment. PATIENT: The case was a 77-year-old man with non-insulin dependent diabetes mellitus, who presented with otalgia and purulent otorrhea. Intervention was with surgical debridement, antifungal agents, and hyperbaric oxygen treatment. The main outcome measures were radiological and histological findings. CONCLUSIONS: A. flavus is a rare cause of malignant otitis externa. Aggressive treatment should include surgical debridement, with appropriate antifungal agents and hyperbaric oxygen therapy. PMID- 18508402 TI - [Morphometric vectorial method of analysis of the frontal sinuses]. AB - The frontal sinuses are pneumatic cavities located in the thickness of the squama frontalis, which communicate with the nasal cavity through the frontonasal duct. These cavities develop by the pneumatisation extent of some anterior ethmoidal cells. Morphologically, there is a large variability of the frontal sinus shape, size and extent, the position of the intersinusal septum, the existence and number of intrasinusal septa. There exist morphologically atypical frontal sinuses as: uni- or bilateral frontal sinuses aplasia, supernumerary sinuses, great extent of the cavities. Paranasal sinuses can be explored by different methods, but the most accessible and easy to perform is conventional radiological imaging. The radiographs can be morphometrically assessed in order to prove the individuality of these air cavities. PMID- 18508403 TI - Immune regulation and control of regulatory T cells by OX40 and 4-1BB. AB - The TNFR family members OX40 (CD134) and 4-1BB (CD137) have been found to play major roles as costimulatory receptors for both CD4 and CD8 T cells. In particular, in many situations, they can control proliferation, survival, and cytokine production, and hence are thought to dictate accumulation of protective T cells during anti-viral and anti-tumor responses and pathogenic T cells during autoimmune reactions. As opposed to simply controlling the activity of naive, effector, and memory T cells, recent data have suggested that both molecules are also instrumental in controlling the generation and activity of so-called regulatory or suppressor T cells (Treg), perhaps in both positive and negative manners. Part of the action on Treg might function to further promote protective or pathogenic T cells, but alternate activities of OX40 and 4-1BB on Treg are also being described that suggest that there might be control by these molecules at multiple levels that will alter the biological outcome when these receptors are ligated. This review specifically focuses on recent studies of regulatory T cells, and regulatory or suppressive activity, that are modulated by OX40 or 4 1BB. PMID- 18508405 TI - Plant ARGONAUTES. AB - ARGONAUTE (AGO) proteins are integral players in all known small RNA-directed regulatory pathways. Eukaryotes produce numerous types of small RNAs, such as microRNAs (miRNA), small interfering RNAs (siRNA), PIWI-interacting RNAs (piRNAs), scanRNAs and 21U-RNAs, and these RNA species associate with different types of AGO family members, such as AGO, PIWI and group 3 proteins. Small RNA guided AGO proteins regulate gene expression at various levels, including internal genomic DNA sequence elimination (in ciliates), translational repression (animals), and RNA cleavage (all eukaryotes), which in some cases is followed by DNA methylation and chromatin remodeling. The plant model species Arabidopsis contains ten AGO proteins belonging to three phylogenetic clades. This review covers our current knowledge of plant AGO functions during miRNA- and siRNA mediated regulation of development and stress responses, siRNA-mediated antiviral immune response, and siRNA-mediated regulation of chromatin structure and transposons. PMID- 18508404 TI - Targeting tumors with LIGHT to generate metastasis-clearing immunity. AB - Metastatic diseases cause the majority of morbidity and mortality of cancer patients. Established tumors form both physical and immunological barriers to limit immune detection and destruction. Current immunotherapy of vaccination and adoptive transfer shows limited effect at least in part due to the existing barriers in the tumors and depending on the knowledge of tumor antigens. Tumor necrosis factor (TNF) superfamily (TNFSF) member 14 (TNFSF14) LIGHT interacts with stromal cells, dendritic cells (DCs), NK cells, naive and activated T cells and tumor cells inside the tumor tissues via its two functional receptors, HVEM and lymphotoxin beta receptor (LTbetaR). Targeting tumor tissues with LIGHT leads to augmentation of priming, recruitment, and retention of effector cells at tumor sites, directly or indirectly, to induce strong anti-tumor immunity to inhibit the growth of primary tumors as well as eradicate metastases. Intratumor treatment would break tumor barriers and allow strong immunity against various tumors without defining tumor antigens. This review summarizes recent findings to support that LIGHT is a promising candidate for an effective cancer immunotherapy. PMID- 18508406 TI - Prevalence survey of schistosomiasis in Mindanao and the Visayas, The Philippines. AB - The first two phases of a national prevalence survey of schistosomiasis in The Philippines were completed in Mindanao in 2005 and the Visayas in the first quarter of 2007. The design was a stratified two-step systematic cluster sampling, with two Kato-Katz thick smears examined from each participant. In Mindanao, a total of 22 provinces spread in six regions were covered by the survey with five barangays (equivalent to a village) per province for a total of 110 barangays. The response rate was 70.9% with a total of 21,390 individuals examined. The province of Maguindanao, a known endemic area for schistosomiasis japonica, failed to take part in the survey. In the Visayas, 10 out of 11 provinces, spread out in three regions, participated in the survey. There were 6321 respondents for an overall participation rate of 32.2%. Mindanao showed a wider coverage of the disease than the Visayas (60% versus 45%). By region, Caraga or Region 13 ranked first in Mindanao and Region 8 in the Visayas. By province, Agusan del Sur is first on the list, followed by Northern Samar and then Eastern Samar. Overall, the prevalence rate among males is higher than that of females suggesting the occupational hazard of farming and fishing among the males. The higher exposure among farmers and fishermen is also borne out by the age distribution of the disease. Prevalence remains consistently high among the adults compared with the younger age groups. The survey also covered other helminth infections that can be detected in a stool survey, notably soil transmitted helminthes and food-borne trematodes. PMID- 18508407 TI - Distribution of filarial elephantiasis and hydrocele in Matara district, Sri Lanka, as reported by local leaders, and an immunological survey in areas with relatively high clinical rates. AB - To eliminate lymphatic filariasis by means of mass drug administration, it is essential to have reliable data on the disease distribution and prevalence in targeted areas. In Matara district, Sri Lanka, self-administered questionnaires were mailed to 2105 local leaders questioning the presence and the numbers of elephantiasis and hydrocele cases. The information provided by them revealed that elephantiasis was clearly aggregated in the southern part of the district along the coast, while hydrocele was distributed rather evenly in the whole district, including Deniyaya region where no endemic filariasis had been known. To confirm active transmission of filariasis in Deniyaya, Wuchereria bancrofti antigen and filaria-specific urinary IgG4 antibody were measured with 2436 subjects. The positive rates for antigen and antibody were 0.6% and 4.3%, respectively. The titer analysis of IgG4 according to age revealed that the youngest IgG4 positive was 3 years old, and that in 10 years old or less, there were 16 positives out of 607 children examined (2.6%). It was concluded that filarial transmission at a low level was going on in the region. PMID- 18508408 TI - Non-attendees' attitudes to the design of a cardiac rehabilitation programme focused on information of risk factors and professional involvement. AB - BACKGROUND: Enhancing the accuracy of the content of cardiac rehabilitation programmes (CRPs) and providing the mediators preferred by patients can increase attendance rates in line with secondary prevention goals. The aim of this study was therefore to explore non-attendees' attitudes to the design of a CRP focused on information of risk factors and professional involvement. METHOD: Consecutive patients with coronary heart disease (n=100) who declined to participate in a CRP answered a questionnaire focusing on patients' attitudes to risk factors and cardiac rehabilitation. RESULTS: Non-attendees considered that information of hypertension and information of sedentary lifestyle constituted the most important content of a CRP. Physicians, nurses and social workers were considered the most suitable professional categories. Females preferred nurses when discussing smoking issues while males preferred physicians. More males compared to females preferred occupational therapists for dealing with stress, physicians for depression and social workers for social isolation. CONCLUSIONS: Non attendees' attitudes are similar to those of attendees and quite traditional in that they favour physician or nurse-led activities. There is a difference in attitude between male and female. PMID- 18508409 TI - T-cell responses in osteosarcoma patients vaccinated with an anti-idiotypic antibody, 105AD7, mimicking CD55. AB - We assessed T-cell responses in young osteosarcoma patients vaccinated with 105AD7, 1-6 months after having received chemotherapy. 105AD7 is a human anti idiotypic antibody mimicking CD55, a glycoprotein that protects from attack by complement and which is overexpressed on osteosarcoma cells. Seven out of 21 investigated patients made a IFN-gamma T-cell response against the vaccine, 105AD7 as assessed by ELISPOT. Cytokine secretion was analysed using Luminex assays and revealed TNF-alpha and GM-CSF responses not only to the vaccine but also towards the native antigen, CD55, in 5 / 14 (36%) of investigated patients. Importantly, the Luminex assay was found to be more sensitive than the more established T-cell assays (ELISPOT and proliferation assay), since responses towards the native antigen were recorded in this assay. Clinical responses and induction of immune responses to both the anti-idiotype and the native CD55 antigen support the use of CD55 as a target in cancer treatment. PMID- 18508410 TI - Demonstration of autoantibody binding to muscarinic acetylcholine receptors in the salivary gland in primary Sjogren's syndrome. AB - A significant pathogenetic role of antimuscarinic acetylcholine receptor-3 (anti m3AChR) autoantibodies in primary Sjogren's syndrome (pSS) has been suggested. However, the binding of these antibodies to the receptors in the target tissues has not yet been demonstrated. In this study, the binding characteristics of pSS sera and anti-m3AChR-monospecific sera affinity-purified from pSS patients to labial salivary gland samples from healthy subjects were studied with light- and electron microscopy. Furthermore, the ultrastructural localisation of in vivo deposited antibodies in pSS salivary glands was also investigated. Light microscopic immunohistochemistry revealed the binding of the anti-m3AChR-specific sera to the membrane of acinar cells. Similar reaction end-products were observed in the pSS salivary gland epithelial cell membranes. With electron microscopy, the autoantibody binding was observed to be localised to the junctions of epithelial cell membranes with nerve endings, both in normal and pSS glands. The results indicate that anti-m3AChR antibodies bind to the receptors in the salivary glands. PMID- 18508411 TI - Mouth breathing increases the pentylenetetrazole-induced seizure threshold in mice: a role for ATP-sensitive potassium channels. AB - Nasal obstruction and consequent mouth breathing have been shown to change the acid-base balance, producing respiratory acidosis. Additionally, there exists a large body of evidence maintaining that acidosis affects the activity of ATP sensitive potassium (K(ATP)) channels, which play a crucial role in the function of the central nervous system (CNS), for example, in modulating seizure threshold. Thus, in the study described here, we examined whether mouth breathing, induced by surgical ligation of nostrils, could affect the seizure threshold induced by pentylenetetrazole in male NMRI mice. Using the selective K(ATP) channel opener (diazoxide) and blocker (glibenclamide), we also evaluated the possible role of K(ATP) channels in this process. Our data revealed that seizure threshold was increased 6 to 72 hours after nasal obstruction, reaching a peak 48 hours afterward, compared with either control or sham-operated mice (P<0.01). There was a significant decrease in pH of arterial blood samples and increase in CO(2) partial pressure (PCO(2)) during this time. Systemic injection of glibenclamide (1 and 2mg/kg, ip, daily) significantly prevented the increase in seizure threshold in 48-hour bilaterally nasally obstructed mice, whereas it had no effect on seizure threshold in sham-operated mice. Systemic injection of diazoxide (25mg/kg, ip, daily) had no effect on seizure threshold in all groups, whereas higher doses (50 and 100mg/kg, ip, daily) significantly increased seizure threshold in both 48-hour-obstructed and sham-operated mice. The decrease in seizure threshold induced by glibenclamide (2mg/kg, ip, daily) was prevented by diazoxide (25mg/kg, ip, daily). These results demonstrate for the first time that mouth breathing, which could result in respiratory acidosis, increases seizure threshold in mice and K(ATP) channels may play a role in this effect. PMID- 18508412 TI - Purification and characterization of glutathione reductase from rainbow trout (Oncorhynchus mykiss) liver and inhibition effects of metal ions on enzyme activity. AB - Glutathione reductase (E C: 1.8.1.7; GR) was purified from rainbow trout (Oncorhynchus mykiss) liver, and some characteristics of the enzyme were investigated. The purification procedure consisted of four steps: preparation of homogenate, ammonium sulfate fractionation, affinity chromatography on 2',5'-ADP Sepharose-4B and gel filtration chromatography on Sephadex G-200. The enzyme, with a specific activity of 27.45 U/mg protein, was purified 1,654-fold with a yield of 41%. Optimal pH, stable pH, optimal temperature, optimum ionic strength, molecular mass, KM and Vmax values for GSSG and NADPH were also determined for the enzyme. In addition, Ki values and inhibition types were determined for GSH and NADP+. Additionally, inhibitory effects of metal ions (Cd+2, Cu+2, Pb+2, Hg+2, Fe+3 and Al+3) on glutathione reductase were investigated. Ki constants and IC50 values for metal ions were determined by Lineweaver-Burk graphs and plotting activity % vs. [I], respectively. IC50 values of Cd+2,Cu+2, Pb+2, Hg+2, Fe+3 and Al+3 were 0.0655, 0.082, 0.122, 0.509, 0.797 and 0.804 mM, and the Ki constants for Cd+2 and Cu+2 were 0.104+/-0.001, 0.117+/-0.001, respectively. PMID- 18508413 TI - Hypoxia inhibits fish spawning via LH-dependent final oocyte maturation. AB - To evaluate the effects of long term hypoxia exposure on fish spawning, mature common carp, Cyprinus carpio carpio (Linnaeus) were subjected to either normoxia (7.4+/-0.2 mgO(2)mg O(2) L(-1)) or hypoxia (1.0+/-0.2 mgO(2)O(2) L(-1)) for more than two months. Gonadosomatic index (GSI), and concentrations of serum luteinizing hormone (LH), testosterone (T), and estroldiol (E2) were measured and gonad histology examined. Hypoxia inhibits fish spawning even though the gonad and oocytes developed under hypoxia exposure. LH levels of female carp were significantly decreased upon chronic exposure to hypoxia, and the final oocyte maturation in hypoxic females was significantly retarded. The results indicated that hypoxia may inhibit fish spawning through LH-dependent final oocyte maturation. In addition, no courtship was observed in hypoxic males. In conclusion, hypoxia impairs fish ovulation and, therefore, spawning and reproduction. LH levels were reduced leading to a failure of oocyte maturation. This, along with a lack of courtship by males may be the major mechanisms involved in hypoxic inhibition of reproduction in carp. PMID- 18508414 TI - Uptake of oleoyl-chitosan nanoparticles by A549 cells. AB - The aim of the present study was to evaluate cellular uptake of oleoyl-chitosan (OCH) nanoparticles by using A549 cells, a human lung carcinoma cell line, for drug and gene delivery applications. In this study, self-assembled OCH nanoparticles encapsulating a fluorescent marker molecule, fluorescein isothiocyanate (FITC), were prepared and characterized. The effects of particle size, concentration, and incubation time on the cellular uptake of the nanoparticles (FITC-OCH nanoparticles) were quantified by spectrofluorometric measurement and confirmed using fluorescence microscopy studies. The nanoparticles were taken up by the cells, and levels of binding and uptake increased with the decrease of particle size and the increase of particle concentration and incubation time. These results implied that the OCH nanoparticles have great potential to be applied as a drug carrier system to deliver drugs into the cells. PMID- 18508415 TI - Variability of hemagglutinin-neuraminidase and nucleocapsid protein of vaccine and wild-type mumps virus strains. AB - The mumps virus (MuV) molecular evolution is characterized by the co-circulation of numerous distinct strains. Standardized phylogenetic analyses based on the nucleotide sequences of the SH gene are important for mumps surveillance, but lack the information regarding antigenic properties. So far, the location of antigenic epitopes has been determined for two MuV proteins, the hemagglutinin neuraminidase (HN) and the nucleocapsid (N) protein. We performed multiple sequence comparisons of putative HN and N protein sequences in order to describe their diversity and plasticity, and to determine the level of similarity between vaccine and wild-type strains. The results of full-length HN or N protein phylogeny showed that MuV strains form a number of differing clades which are in concordance with grouping obtained by standard MuV genotyping. When vaccine strains are compared to all wild-type strains, the highest mean percentage of amino acid differences in both HN and N protein analysis was found for Jeryl Lynn 5 and Jeryl Lynn 2 strains while the lowest value was obtained for Leningrad-3 and L-Zagreb strains. When only 3 antigenic regions of the HN protein, comprising 45 amino acids in total, were investigated, the diversity is considerably diminished: 51.5% of all putative HN proteins show identical sequences (including those of vaccine strains L-Zagreb, Leningrad-3, Hoshino and Urabe). Another 26.5% proteins (including Miyahara vaccine strain) differ in only one amino acid, while the others differ in two to five amino acids from the most common sequence. Jeryl Lynn 2 and Jeryl Lynn 5 strains differ in four amino acids each. N protein antigenic sites have been mapped within its hypervariable C-terminus. Our results indicate that there might be genotype-specific amino acids residing in this antigenic region. The results of our study present the background information for investigations of MuV heterogeneity and antigenic diversity. PMID- 18508416 TI - A simple bioanalytical assay for determination of montelukast in human plasma: application to a pharmacokinetic study. AB - An analytical method based on high-performance liquid chromatographic (HPLC) was developed for the determination of montelukast in human plasma using mefenamic acid as an internal standard. After precipitation of plasma proteins with acetonitrile, chromatographic separation was carried out using a Zorbax Eclipse XDB C8 (150 mm x 4.6 mm i.d., 5 microm) with mobile phase consisted of methanol acetonitrile-0.04M disodium hydrogen orthophosphate (22:22:56, v/v, pH 4.9). The wavelengths of fluorescence detection were set at 350 nm for excitation and 450 nm for emission. The linearity was confirmed in the concentration range of 5-1000 ng/ml in human plasma. Intra- and inter-day accuracy determined from quality control samples were 101.50 and 107.24%, and 97.15 and 100.37%, respectively. Intra- and inter-day precision measured as coefficient of variation were < or =4.72 and < or =9.00%, respectively. Extraction recoveries of drug from plasma were >48.14%. The protocol herein described was employed in a pharmacokinetic study of tablet formulation of montelukast in healthy Thai male volunteers. PMID- 18508417 TI - Quantification of erufosine, the first intravenously applicable alkylphosphocholine, in human plasma by isotope dilution liquid chromatography tandem mass spectrometry using a deuterated internal standard. AB - A sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of erucylphosphohomocholine (erufosine, ErPC(3)) in pharmacokinetic studies. Nine-fold deuterated ErPC(3) was used as the internal standard. Following protein precipitation, reversed phase chromatography was performed. For analyte detection, electrospray ionization in the positive mode was applied. The mass transition m/z 504.4>139.1 was recorded for ErPC(3), and the transition m/z 513.7>139.1 for the internal standard, respectively. Good linearity with a correlation coefficient >0.99 was found for the range of 0.48-15 mg/L ErPC(3) in plasma (0.93-29.8 microM), the important range for clinical pharmacokinetic analysis. Interassay coefficients (n=10) of variation between 4.2% and 5.5% were found for ErPC(3) pool samples with concentrations between 4.7 mg/L and 44.0mg/L, respectively. The method has been used for analyses during a phase I clinical trial of ErPC(3). PMID- 18508418 TI - The role of resuscitation drugs and placental transfusion in the delivery room management of newborn infants. AB - Medications are used rarely in newborn resuscitations and are probably justifiable in less than 0.1% of births. Doses used are mainly extrapolated from animal and adult data. Despite this, the drugs used, their order and route of administration have all been sources of controversy for many years. There have been polarised views, often focusing upon adrenaline and sodium bicarbonate and more recently new drugs such as vasopressin have been suggested, once again extrapolating from adult experience. This article examines the sparse data behind the use of any medication at birth and the poor outcome data available. The appropriate decline in the indiscriminate use of volume expansion is considered and balanced by the increasing evidence in favour of delayed clamping of the umbilical cord. Focusing on the basic steps of resuscitation, improving the quality of their application and avoiding relative hypovolaemia, must improve the quality of outcome data. The place of medications in newborn resuscitation should be regarded as experimental and still requires evidence to justify their use especially in premature babies. PMID- 18508419 TI - Expression of androgen, estrogen and progesterone receptors in mucinous carcinoma of the breast. AB - Hormone receptors play important roles in breast cancer. We investigated the expression of hormone receptors in breast cancer to evaluate the importance of hormone receptors in the clinicopathology of breast cancer. Androgen receptor (AR), estrogen receptor (ER) and progesterone receptor (PR) expression characteristics were evaluated using immunohistochemistry stain, comparing patient age, tumor size and axillary lymph node status for 23 pure mucinous and 105 non-mucinous infiltrating ductal carcinomas in the human female breast. Mucinous carcinoma with axillary lymph node metastasis occurred less frequently than non-mucinous carcinoma (11.8% vs. 55.2%; p = 0.01). Compared with the non mucinous type, mucinous carcinoma specimens showed less AR expression (21.7% vs. 51.4%; p = 0.01) but more ER expression (78.3% vs. 52.4%; p = 0.02). In addition, AR expression was also associated with ER and/or PR coexpression (37/74, 50%) in infiltrating ductal carcinoma. But only three of 20 (15%) mucinous carcinoma specimens with AR expression had associated ER and/or PR coexpression. Our findings revealed that mucinous carcinoma samples from the breast show distinct clinicopathologic and hormone receptor expression features compared to non mucinous carcinoma. PMID- 18508420 TI - Correlation between quadrant specific automatic visual field defect and retinal nerve fiber layer thickness as measured by scanning laser polarimetry in patients with primary open angle glaucoma. AB - The purpose of this study was to correlate quadrant specific Humphrey visual field mean deviation (MD) with retinal nerve fiber layer (RNFL) thickness as measured by scanning laser polarimetry (GDx), and to determine whether there is a difference in the correlation with visual field defect between the Asian normative database provided by GDx (GDx database) and our native normative database (KMU database). In an age-matched study, a control group of 240 normal eyes underwent GDx. Another 60 eyes with visual field defect due to primary angle glaucoma underwent autoperimetry and GDx examination. First, we compared four GDx measurements between the control and study groups. Next, we divided the visual field into four quadrants (superior, inferior, temporal, nasal) and calculated the MD of each quadrant. We correlated the MD of superior, inferior and overall visual field with RNFL thickness judged by two databases (the GDx Asian internal normative database and the database from our control group). GDx detected abnormal RNFL thickness significantly more accurately when using the KMU database (p = 0.0473 for superior quadrant; p = 0.0074 for inferior quadrant; p = 0.0011 for average thickness) than when using the GDx database. There was no significant difference in the specificity between these two databases. The normal ranges in the GDx internal normative database for Asians are too wide. By using our own GDx normative database, the correlations with MD of autoperimetry were significantly improved. We suggest that every laboratory and clinic establish its own normative database of GDx in Asia. PMID- 18508421 TI - Association among metabolic syndrome, testosterone level and severity of erectile dysfunction. AB - The purpose of this study was to determine the influence of metabolic syndrome (MS) and serum testosterone in patients with erectile dysfunction (ED) and their possible association. A total of 103 men with ED were enrolled. The International Index of Erectile Function (IIEF) questionnaire was used to assess erectile condition. MS was defined according to the criteria formulated by the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) and the International Diabetes Federation (IDF). The mean age of the study population was 57.5 +/- 10.7 years, with an average IIEF of 14.7 +/- 6.7. The age and prevalence of MS using the NCEP ATP III criteria, but not the IDF criteria, were significantly different between mild and moderate/severe ED patients (p = 0.031 and 0.009, respectively). The percentage of hypertension (78.6% vs. 36.2%; p < 0.001) and raised fasting glucose levels (46.4% vs. 19.1%; p = 0.004) were significantly higher in the moderate/severe ED group, and both differences remained significant in multivariate analysis (p = 0.001 and 0.042, respectively). In addition, serum testosterone levels were significantly lower in ED patients with MS (p = 0.002). In summary, the presence of MS is associated with more severe ED. Among the components of MS, elevated blood pressure and fasting blood glucose were independent risk factors. NCEP ATP III criteria seem to correlate better with the degree of ED than the IDF definition. Our results also indicate that MS is associated with a lower testosterone level in patients with ED. PMID- 18508422 TI - Characteristics of febrile patients with normal white blood cell counts and high C-reactive protein levels in an emergency department. AB - Fever is one of the more common chief complaints of patients who visit emergency departments (ED). Many febrile patients have markedly elevated C-reactive protein (CRP) levels and normal white blood cell (WBC) counts. Most of these patients have bacterial infection and no previous underlying disease of impaired WBC functioning. We reviewed patients who visited our ED between November 2003 and July 2004. The WBC count and CRP level of patients over 18 years of age who visited the ED because of or with fever were recorded. Patients who had normal WBC count (4,000-10,000/L) and high CRP level (> 100 mg/L) were included. The data, including gender, age and length of hospital stay, were reviewed. Underlying diseases, diagnosis of the febrile disease and final condition were recorded according to the chart. Within the study period, 54,078 patients visited our ED. Of 5,628 febrile adults, 214 (3.8%) had elevated CRP level and normal WBC count. The major cause of febrility was infection (82.24%). Most of these patients were admitted (92.99%). There were 32 patients with malignant neoplasm, nine with liver cirrhosis, 66 with diabetes mellitus and 11 with uremia. There were no significant differences in age and gender between patients with and those without neoplasm. However, a higher inhospital mortality rate and other causes of febrility were noted in patients with neoplasm. It was not rare in febrile patients who visited the ED to have a high CRP level but normal WBC count. These patients did not necessarily have an underlying malignant neoplasm or hematologic illness. Factors other than malignant neoplasm or hematologic illness may be associated with the WBC response, and CRP may be a better indicator of infection under such conditions. PMID- 18508423 TI - Endothelin-1 enhances corneal fibronectin deposition and promotes corneal epithelial wound healing after photorefractive keratectomy in rabbits. AB - The objective was to study the effects of endothelin-1 (ET1) on corneal wound healing after photorefractive keratectomy (PRK) in rabbit corneas. Following PRK, 18 New Zealand white rabbits were treated with ET1 in the right eyes and with phosphate-buffered salt solution (PBS) in the left eyes. Corneal epithelial wound size, corneal haze and corneal thickness were recorded. Corneal extracellular matrixes, including collagen types 3, 4 and 7, chondroitin sulfate and fibronectin, were investigated using immunohistochemistry study. ET1 increased the rate of healing of corneal epithelial wounds in rabbits. Anti-fibronectin fluorescence was present at week 12 and week 24 in ET1-treated eyes but not in the control eyes. There were no significant differences in corneal haze, corneal thickness and changes in other extracellular matrixes between ET1- and PBS treated eyes. ET1 can enhance the deposition of fibronectin in corneal stroma and promote corneal epithelial wound healing after PRK. The increase in fibronectin probably explains the increased healing rate of corneal epithelial wounds. PMID- 18508424 TI - Low-grade astrocytoma associated with abscess formation: case report and literature review. AB - A rare case of low-grade astrocytoma associated with abscess formation occurred in a 52-year-old man presenting with Broca's aphasia. He underwent craniotomy and tumor removal under the impression of brain tumor with necrotic cystic change. Abscess accumulation within the intra-axial tumor was found intraoperatively. Literature related to brain abscess with brain tumor is reviewed, with an emphasis on abscesses with astrocytoma. We discuss the common brain tumors that are associated with abscess, pathogens that coexist with brain tumor, and the pathogeneses of coexisting brain abscess and tumor. It is very important to know how to differentiate between and diagnose a brain abscess and tumor, or brain abscess with tumor, preoperatively from clinical presentation and through the use of computed tomography, conventional magnetic resonance imaging, diffusion weighted imaging or magnetic resonance spectroscopy. PMID- 18508425 TI - Delayed surgery in a complex trauma patient with aortic rupture, blunt head trauma and liver laceration: a case report. AB - Aortic rupture is a critical condition in trauma patients. Most patients are killed at the scene of the accident. A patient who survives long enough to reach hospital also has a high risk of rupture during management. We report a patient who was transferred from a municipal hospital with the emergency complex of blunt head trauma and chest and abdominal contusion. Chest computed tomography scan revealed aortic rupture. Liver laceration with hemodynamic stability and brain concussion were also diagnosed in the emergency room. She was admitted to the trauma intensive care unit without emergency surgery. She received aorta repair after 5 days of observation. After the operation, the patient recovered very well. Delayed surgery for aortic rupture as a treatment choice may be of benefit in selected complex trauma cases. PMID- 18508426 TI - Primary urethral plasmacytoma: a case report and literature review. AB - Plasmacytomas of the urethra are extremely rare neoplasias; they may occur as isolated tumors or concomitantly with generalized multiple myeloma. Herein, we describe the clinical presentation and characteristics of a patient with primary plasmacytoma of the urethra. A 51-year-old man presented with terminal hematuria and a palpable penile mass. Magnetic resonance urethrography revealed a 3-cm long stenotic segment along which the urethral mucosa was found to be irregular. On urethroscopy, papillary mucosal projections extending to the presphincteric area were noted. Lesions were found to be composed primarily of neoplastic plasma cells capable of producing mainly lambda light chain. Upon diagnosis, the patient received external beam radiation therapy targeting the pelvic region. The lesion diminished in size progressively during the treatment course. He was disease-free after 6 months. Although it is a relatively rare disease, primary urethral plasmacystoma should be considered in the differential diagnosis of urethral tumors and radiation therapy should be an integral part of the treatment strategy. PMID- 18508427 TI - Non-substrate peptides influencing dipeptidyl peptidase IV/CD26 activity and immune cell function. AB - Investigations using inhibitors of dipeptidyl peptidase IV (DP IV) activities and DP IV-/- mice indicated an immunoregulatory role of DP IV that could not be compensated by DP IV-like enzymes. The HIV-1 Tat protein was identified as the first natural inhibitor of DP IV and as immunosuppressor. This review summarizes our investigations on the identification of the amino acid motif of Tat responsible for DP IV inhibition and of endogenous DP IV-inhibitory ligands that suppress immune cell activation. Examinations on numerous peptides carrying the N terminal Xaa-Xaa-Pro motif of Tat revealed that tryptophan at position two strongly enhanced DP IV inhibition and immunosuppression. Here, we present evidence that the thromboxane A2 receptor exposing N-terminal Met-Trp-Pro at the cell surface could be a potential endogenous, inhibitory DP IV ligand. Moreover, our data suggest that the major envelope proteins p37k of the orthopoxviruses variola virus and vaccinia virus, as well as the B2L antigen of the parapoxvirus orf, that also carry N-terminal Met-Trp-Pro, could mediate immunosuppressive effects. Further examinations are in progress to identify new physiologic, inhibitory DP IV ligands and to enlighten the mechanism underlying the DP IV mediated effects. PMID- 18508428 TI - Immuno-gene therapy approaches for cancer: from in vitro studies to clinical trials. AB - Immunotherapy against cancer basically aims at either broadly stimulating the immune system or at engineering an immune response against a targeted tumor associated antigen (TAA). In this review, we focus on the translation of immuno gene therapy strategies into clinical trials for various cancers. Rather than being an exhaustive compendium of the literature, the focus of this article is to underline how anti-cancer immunotherapy strategies have evolved recently. Previously, studies have used different vectors to either express immuno stimulatory molecules or a targeted TAA. Investigators are now directing efforts to both target a TAA and to stimulate the immune system by direct or viral administration of cytokines or co-stimulatory molecules. Some groups have also tried to combine genetic immunotherapy with chemotherapy, and results have been encouraging. This novel concept might open new perspectives for the treatment of patients with advanced-stage cancer. PMID- 18508430 TI - Biochemistry of inositol lipids. AB - Nature has created an immense combinatorial and structural heterogeneity among lipids. It is becoming increasingly accepted that the vast range of unique chemical entities encodes for distinct functions within biological systems. A unique group of lipids which stands out in terms of diversity as well as biological activity are inositol-containing lipids. The most well characterized inositol lipids are the phosphoinositides, phosphorylated derivatives of glycerophosphoinositol, which play a wide variety of cellular roles in many eukaryotic cells. Less well understood are ceramides containing inositol in fungi, and inositol glycolipids in pathogens. Here we review biochemical aspects of inositol-containing lipids with a focus on novel analytical procedures for their characterization. PMID- 18508431 TI - Diagnostics of pathogenic splicing mutations: does bioinformatics cover all bases? AB - Pathogenic splicing alterations caused by point mutations in both splice sites and auxiliary cis-regulatory elements are increasingly recognized as an important mechanism through which gene mutations cause human disease. Unfortunately, in routine genetic diagnostic settings, splicing mutations may escape identification, due to the lack of RNA samples. Since most patients are genotyped only, any computational prediction of mutation effects on splicing can be beneficial for the human geneticist. Here, we review common techniques to identify human point mutations and delineate the molecular basis for splice site recognition. Moreover, this article provides basic insights into web-tools predicting splice sites and cis-regulatory elements and discusses their benefits for judgment of clinically identified sequence variants of disease-specific genes. PMID- 18508429 TI - Gingipain-dependent interactions with the host are important for survival of Porphyromonas gingivalis. AB - Porphyromonas gingivalis, a major periodontal pathogen, must acquire nutrients from host derived substrates, overcome oxidative stress and subvert the immune system. These activities can be coordinated via the gingipains which represent the most significant virulence factor produced by this organism. In the context of our contribution to this field, we will review the current understanding of gingipain biogenesis, glycosylation, and regulation, as well as discuss their role in oxidative stress resistance and apoptosis. We can postulate a model, in which gingipains may be part of the mechanism for P. gingivalis virulence. PMID- 18508432 TI - The type I insulin-like growth factor receptor pathway: a key player in cancer therapeutic resistance. AB - The insulin-like growth factor (IGF) ligands stimulate cellular proliferation and survival by activating the type I insulin-like growth factor receptor (IGF-IR). As a result, the IGF signaling system is implicated in a number of cancers, including those of the breast, prostate, and lung. In addition to mitogenic and anti-apoptotic roles that may directly influence tumor development, IGF-IR also appears to be a critical determinant of response to numerous cancer therapies. This review describes the role of the IGF-IR pathway in mediating resistance to both general cytotoxic therapies, such as radiation and chemotherapy, and targeted therapies, such as tamoxifen and trastuzumab. It concludes with a description of approaches to target IGF-IR and argues that inhibition of IGF signaling, in conjunction with standard therapies, may enhance the response of cancer cells to multiple modalities. PMID- 18508433 TI - Oxidative stress in neurodegeneration and available means of protection. AB - Substantial pieces of direct and indirect evidence have mounted over the years linking the induction of oxidative stress to a plethora of disease conditions, not least those associated with the death of neurons. The causal relationship between oxidative damage and neurodegeneration is, however, not yet clear and still a subject of intense investigation. Nevertheless, the phenomenon of oxidative neuronal death has received considerable attention in a frantic search for efficacious therapies for the management of neurological and neurodegenerative conditions. The redox-active nature of reactive oxygen species (ROS), which in their excessive levels induce oxidative stress, the prevalence of ROS production in biological systems, the complexity of interrelationships among these species, and the context-dependent adequacy and resilience of the antioxidant defense systems are some of the challenges that basic research has to grapple with to advance successfully to the translational stage. Much still has to be understood for research efforts in this field to yield enduring therapies. In this review, we examine the nature (chemistry) of ROS, the relationships between them, their physiological functions, the roles of oxidative stress in neurodegeneration, the mechanisms of cell death induced by oxidant species, and the available means of protecting neurons against oxidative damage. PMID- 18508434 TI - Chemokines in lupus nephritis. AB - Lupus nephritis is a common solid organ manifestation of systemic lupus erythematosus (SLE). The disease is tightly linked to the production of autoantibodies and circulating immune complexes, i.e. immune complex glomerulonephritis. In this process chemokines mediate multiple biological effects, e.g. orchestrating proinflammatory microenvironments, the recruitment of immune cell subsets into the kidney, as well as the local activation of such immune effector cells. Autoimmune mice with targeted deletions of certain single chemokines or chemokine receptors are protected from renal autoimmune tissue injury. Interventional studies with specific antagonists against certain chemokines and chemokine receptors further support these findings. In this review we summarise the available experimental and human data on the expression and functional role of chemokines and chemokine receptors in lupus nephritis. PMID- 18508435 TI - Heterotrimeric G-proteins in plant development. AB - Signaling through heterotrimeric G-proteins (G-proteins) is a conserved mechanism found in all eukaryotes. In plants, the repertoire of G-protein signaling complex is much simpler than in metazoans. Specifically, the genome of the model plant, Arabidopsis, encodes only one canonical Galpha, one Gbeta, and two Ggamma subunits. Similarly, only one Regulator of G-protein Signaling (RGS) protein is encoded by the Arabidopsis genome, and no bona fide G-protein-coupled receptor (GPCR) together with its ligand has been unequivocally identified. Nonetheless, several proteins, including AtPIRIN1, PLDa 1, PD1, and THF1, have been shown to physically interact with the Arabidopsis heterotrimeric G-protein alpha subunit (GPA1), and are potential downstream effectors for GPA1. The smaller repertoire of the heterotrimeric G-protein complex in plants offers a unique advantage over its counterpart in mammals for dissecting their roles in development. The analyses of loss-of-function alleles and gain-of-function transgenic lines of G protein subunits and signaling components suggest that the G-proteins play regulatory roles in multiple developmental processes ranging from seed germination and early seedling development to root development and organ shape determination. Future studies are expected to reveal more components of the heterotrimeric G-protein signal transduction pathways, and to identify the mechanisms by which G-proteins regulate phenotypic and developmental plasticity. PMID- 18508436 TI - Protective and pathologic immune responses against Candida albicans infection. AB - Candida albicans is an important opportunistic fungal pathogen. Clinical observations have indicated that both innate and adaptive immune responses are involved in recovery from initial infection, but analysis in murine models has shown that the contribution of the two arms of the cellular immune response differ in oral, vaginal, and systemic infections. The relative contributions of T cells and phagocytic cells, and the cytokines that mediate their interactions are discussed for each of the different manifestations of the disease, and the consequences of infection, in terms of protection and pathology, are evaluated. PMID- 18508437 TI - G-protein coupled chemoattractant receptors and cancer. AB - Chemoattractant receptors are a group of seven transmembrane, G protein coupled receptors (GPCRs). They were initially identified mainly on leukocytes to mediate cell migration in response to pathogen or host-derived chemotactic factors. During the past decade, chemoattractant GPCRs have been discovered not only to mediate leukocyte chemotaxis thus promoting innate and adaptive host immune responses, but also to play essential roles in development, homeostasis, HIV infection, angiogenesis and wound healing. A growing body of evidence further indicates that chemoattractant GPCRs contribute to tumor growth, invasion, angiogenesis/angiostasis and metastasis. The diverse properties of GPCRs in the progression of malignant tumors have attracted intense interest in their potential as novel anti-tumor pharmacological targets. PMID- 18508438 TI - Gene therapy for esophageal squamous cell carcinoma. AB - Despite improvement of surgical treatment and application of multi-modality therapies to advanced esophageal cancer, the prognosis is extremely poor in patients with T4 tumors. Based on the genetic background of esophageal cancer, we have developed various gene therapy strategies against human esophageal cancer cells. In this article, we reviewed molecular events of esophageal cancer and gene therapy approaches for its treatment. First, we analyzed p53 genetic alterations and angiogenesis in esophageal cancer. Second, we evaluated an impact of p53 recombinant adenoviral vector (Ad5CMV-p53) on esophageal cancer cells. Significant growth suppression was observed following infection with Ad5CMV-p53 in human esophageal squamous cell carcinoma cell lines. This observation suggests that Ad5CMV-p53 may be a potentially effective therapeutic agent for locally advanced esophageal cancer. Promising avenues for investigation include double gene therapy and adjuvant use of gene therapy with radiation therapy. Third, we have performed a clinical study for p53 gene therapy for un-resectable advanced esophageal cancer. This clinical trial was planned to evaluate vector tolerability and efficacy. A total of 10 patients were enrolled into this phase I/II trial. PMID- 18508439 TI - Strategies to expand the living donor pool for kidney transplantation. AB - Structural shortage of deceased donor kidneys for transplantation has resulted in the expansion of living donation programs. A number of possibilities are now being explored, since it became clear that donors do not need to be genetically related to their recipients. Apart from classical direct donation we now conduct paired exchange, list exchange, altruistic donation and domino paired exchange programs. Other alternative programs are desensitization and transplantation across the blood type barrier. The purpose of this article is to give a general view of all optimizing living donation programs by reviewing the literature. First we describe logistic solutions, thereafter the more intensive medical treatments. We observed a wide variation in clinical experiences with living donation dependent on local jurisdiction, culture and customs. Professionals disagree on various ethical issues inherent to alternative programs. In our opinion logistic solutions like paired exchange, list exchange and altruistic donation programs are to be preferred over the more medical demanding programs e.g. desensitization and transplantation across the blood type barrier. PMID- 18508440 TI - Binocular phasic coactivation does not prevent ocular dominance segregation. AB - The segregation of geniculo-cortical afferents into ocular dominance columns is an activity-dependent process. It was hypothesized that this process is susceptible to the temporal patterning of the retinal input. Accordingly, asynchronous activation of the two eyes should enhance ocular dominance segregation but synchronous activation should decrease or prevent it. In order to test the second part of the hypothesis, kitten were raised in strobe light which phasically coactivated the retinal inputs during 10 microsecond flashes at 8 Hz. Strobe rearing prevents retinal motion signals but allows vision of stationary contours. At the age of 10-14 weeks, ocular dominance columns were labeled either transneuronally by (3H)-proline or by (14C)-2-deoxyglucose autoradiography. Contrary to the hypothesis, ocular dominance columns were very well segregated and the pattern closely resembled the pattern observed in squinting cats. We conclude that the light flashes were sufficient to enable binocular competition and that ocular dominance segregation was supported by the mismatch of the stationary contours. Our result thus emphasizes a feature-selective mechanism over mere global temporal patterning of retinal signals. PMID- 18508441 TI - From protein-disease associations to disease informatics. AB - Advancements in high-throughput technology and computational power have brought about significant progress in our understanding of cellular processes, including an increased appreciation of the intricacies of disease. The computational biology community has made strides in characterizing human disease and implementing algorithms that will be used in translational medicine. Despite this progress, most of the identified biomarkers and proposed methodologies have still not achieved the sensitivity and specificity to be effectively used, for example, in population screening against various diseases. Here we review the current progress in computational methodology developed to exploit major high-throughput experimental platforms towards improved understanding of disease, and argue that an integrated model for biomarker discovery, predictive medicine and treatment is likely to be data-driven and personalized. In such an approach, major data collection is yet to be done and comprehensive computational models are yet to be developed. PMID- 18508442 TI - Gene and cell therapy for relapsed leukemia after allo-stem cell transplantation. AB - To control severe GvHD while maintaining strong GvL effects in the context of allo-stem cell transplantation (allo-SCT), a phase I/II clinical trial of infusions of donor lymphocytes transduced with the herpes simplex virus thymidine kinase (TK-DLI) started at the Tsukuba University Hospital. To date, five (2 AML, 2 ALL, and 1 MDS) out of eight patients enrolled in the trial received approximately 7x10(7) transduced cells per kilogram of body weight and four patients showed some clinical responses such inhibition of the leukemic cell proliferation or mitigation of lymph node swelling. Especially, one MDS patient achieved complete remission and has remained in CR for 2 years after the treatment. GvHD developed in two patients (1 acute and 1 chronic) and the acute (grade III) was successfully controlled by administration of ganciclovir without any immunosuppressive drugs. Since HSV-TK as a strong antigen induced CTLs against transduced cells in patients, however, TK-DLI is expected to provide a more effective adoptive immune cell therapy by performance just after allo-SCT where the patient's immune function is severely damaged. PMID- 18508443 TI - Mesenchymal stem cells transplantation protects against rat pulmonary emphysema. AB - Pulmonary emphysema is characterized by loss of alveolar structure. Bone marrow mesenchymal stem cells (MSCs) have been shown to differentiate into alveolar epithelial cells. However, the effect of MSCs transplantation on pulmonary emphysema is unknown. To address this question, cultured bone marrow MSCs from male donor rats were infused into female recipients treated with irradiation and instillation of papain. We found that the emphysematous changes in rats received MSCs transplantation were ameliorated when compared with the rats without MSCs transplantation. Y chromosome fluorescent in situ hybridization (FISH) and immunohistochemical staining for SP-C, confirmed that MSCs engrafted in recipient lungs and differentiated into type II alveolar epithelial cells. Additionally, MSCs transplantation reduced the extent of irradiation and papain-induced alveolar cell apoptosis, likely due to the up-regulation of the expression of Bcl 2 and Bax gene. We conclude that MSCs transplantation protects against the irradiation and papain-induced pulmonary emphysema. The mechanisms of protection may involve the engraftment of MSCs in the lungs, differentiation of MSCs into type II alveolar epithelial cells and suppression of alveolar cell apoptosis. PMID- 18508444 TI - Microglial degeneration in the aging brain--bad news for neurons? AB - We have long promulgated the idea that microglial cells serve an entirely beneficial role in the central nervous system (CNS), not only as immunological sentinels to fend off potentially dangerous infections, but also as constitutively neuroprotective glia that help sustain neuronal function in the normal and especially in the injured CNS when microglia become activated. In recent years, we have reported on the presence of degenerating microglial cells, which are prominent in the brains of aged humans and humans with neurodegenerative diseases, and this has led us to propose a hypothesis stating that loss of microglia and microglial neuroprotective functions could, at least in part, account for aging-related neurodegeneration. In the current review, we sum up the many aspects that characterize microglial activation and compare them to those that characterize microglial senescence and degeneration. We also consider the possible role of oxidative stress as a cause of microglial degeneration. We finish up by discussing the role microglial cells play in terms of amyloid clearance and degradation with the underlying idea that removal of amyloid constitutes a microglial neuroprotective function, which may become compromised during aging. PMID- 18508445 TI - ADR1 interacts with a down-stream positive element to activate PS1 transcription. AB - We have identified downstream promoter sequence of the PS1 gene that may be regulated by novel transcription factors. 3' deletion from +178 to +165 had no effect on PS1 transcription. 3' deletion from +178 to +140 decreased promoter activity by 50%. Further 3' deletion from +178 to +114 decreased promoter activity by 80%. Therefore, a crucial element controlling over 80% of the promoter activity in SK-N-SH cell line is located between +114 and +165. Electrophoretic mobility shift assays suggested that zinc finger proteins Sp1 and ADR1 interacted with the PS1 promoter sequence (+114 to +140) and promoter region (+140 to +165) respectively. A three base pair substitution within the core sequence (GGCGGGGA to GGCGactA) of the ADR1 consensus in the element (+140 to +165) that abolished ADR1-DNA interaction, reduced PS1 transcription by 50%. The substitution mutation in the sequence (+114 to +140) that abolished Sp1-DNA interaction had no effect on PS1 expression. These data suggest that a novel mammalian trans-activator protein ADR1 binds to the downstream element (+140 to +165) to activate PS1 transcription. PMID- 18508446 TI - Soluble NKG2D ligands: prevalence, release, and functional impact. AB - Natural Killer (NK) cells are capable to recognize and eliminate malignant cells. Anti-tumor responses of NK cells are promoted by the tumor-associated expression of cell stress-inducible ligands of the activating NK receptor NKG2D. Current evidence suggests that established tumors subvert NKG2D-mediated tumor immunosurveillance by releasing NKG2D ligands (NKG2DL). Release of NKG2DL has been observed in a broad variety of human tumor entities and is thought to interfere with NKG2D-mediated tumor immunity in several ways. Further, levels of soluble NKG2DL (sNKG2DL) were also found to be elevated under various non malignant conditions, although the functional implications remain largely unclear. Here we review and discuss the available data on the prevalence, release, functional impact, and potential clinical value of sNKG2DL. PMID- 18508448 TI - Involvement of cystatin C in pathophysiology of CNS diseases. AB - Cystatin C Leu68Gln variant is known to induce amyloid deposition in cerebral arterioles, resulting in Icelandic type cerebral amyloid angiopathy (CAA). Wild type cystatin C is also observed in solitary CAA involving amyloid beta protein (Abeta), and accelerates the amyloidogenicity of Abeta in vitro. In neurological inflammatory diseases and leptomeningeal metastasis, low cystatin C levels are accompanied with high activities of cathepsins in the cerebrospinal fluid. Among the cells in CNS, astrocytes appear to secrete cystatin C in response to various proteases and cytokines. Co-localization of Abeta and cystatin C in the brains of Alzheimer's disease (AD) led to the hypothesis that cystatin C is involved in the disease process. We demonstrated that cystatin C microinjection into rat hippocampus induced neuronal cell death in dentate gyrus. Furthermore, apoptotic cell death was observed in neuronal cells treated with cystatin C in vitro. Up regulation of cystatin C was observed in glial cells with neuronal cell death in vivo. These findings indicate the involvement of cystatin C in the process of neuronal cell death. PMID- 18508447 TI - Dietary terpenoids and prostate cancer chemoprevention. AB - Cancer chemoprevention by phytochemicals may be one of the most feasible approaches for cancer control. Phytochemicals obtained from vegetables, fruits, spices, teas, herbs and medicinal plants, such as terpenoids and other phenolic compounds, have been proven to suppress experimental carcinogenesis in various organs in pre-clinical models. Recent studies have indicated that mechanisms underlying chemopreventive potential may be a combination of antioxidant, anti inflammatory, immune-enhancing, and hormone modulation effects, with modification of drug metabolizing enzymes, influence on cell cycle and cell differentiation, induction of apoptosis, suppression of proliferation and angiogenesis playing roles in the initiation and secondary modification stages of neoplastic development. Specific features of prostate cancer, such as high prevalence and long latency period provides ample opportunities for chemopreventive agents to work at various stages of disease progression. Finally, suitable populations with appropriate risk factors, including the presence of pre-malignant lesions and genetic predispositions, need to be well characterized for future chemopreventive interventions. Here we review naturally occurring dietary terpenoids as useful agents for prostate cancer chemoprevention with reference to their classes and sources. PMID- 18508449 TI - Soluble epoxide hydrolase inhibitor, AUDA, prevents early salt-sensitive hypertension. AB - In stroke-prone spontaneously hypertensive rats (SHRSP) end-organ damage is markedly accelerated by high-salt (HS) intake. Since epoxyeicosatrienoic acids (EETs) possess vasodepressor and natriuretic activities, we examined whether a soluble epoxide hydrolase (sEH) inhibitor, 12-(3-adamantan-1-yl-ureido) dodecanoic acid (AUDA), to inhibit the metabolism of EETs, would protect against pathologic changes in SHRSP. Seven-week-old male SHRSP were treated as follows: normal salt (NS), NS + AUDA, HS and HS + AUDA. Systolic blood pressure (SBP) (205 +/- 4 v 187 +/- 7 mmHg) and proteinuria (3.7 +/- 0.2 v 2.6 +/- 0.2 mg/6 h), but not plasma EETs (11.0 +/- 0.9 v 9.7 +/- 1.1 ng/ml), were significantly increased at 9 weeks of age in HS v NS SHRSP. HS was associated with fibrinoid degeneration and hypertrophy of arterioles in the kidney and perivascular fibrosis and contraction band necrosis in the heart. AUDA ameliorated these early salt dependent changes in saline-drinking SHRSP and increased plasma levels of EETs but did not affect water and electrolyte excretion. sEH inhibition may provide a therapeutic strategy for treating salt-sensitive hypertension and its sequelae. PMID- 18508450 TI - Vector-based delivery of siRNAs: in vitro and in vivo challenges. AB - RNA interference (RNAi) induced by small interfering RNAs (siRNAs) has recently become a powerful tool to knock-down gene expression in a sequence-specific manner. In addition to chemically made siRNAs, stable expression of siRNA in the form of short hairpin RNAs (shRNAs) expressed from an RNA polymerase III (pol III) promoter is now widely used approach for the application of RNAi in mammalian cells. However, long-term suppression using constitutive promoters can be problematic and emerging evidence indicates that siRNAs can cause several side effects in human cells. Here we review the recent advances in developing controllable expression vectors in order to accelerate the therapeutic applications of RNAi. PMID- 18508451 TI - Macrophages and cancer. AB - Macrophages are ubiquitous cells physiologically involved in a variety of processes including pathogen destruction, inflammation, tissue repair and remodeling. They have a highly plastic phenotype and their functional polarization is determined by cytokines and factors found within local microenvironments. The role of macrophages during tumor development is ambiguous. At late stages, tumor-associated macrophages are known to produce molecules directly promoting tumor growth, invasion, and metastasis; the so called "myeloid derived suppressor cells" also suppress the adaptive anti-tumor immune response. However, if properly activated, macrophages may control initial tumor development, and pilot studies in cancer patients suggest that adoptive transfers could be beneficial as adjuvant treatment in patients with minimal residual disease. Indeed, a limited tumor mass will probably be insufficient to educate macrophages into a suppressive phenotype. Thus, the macrophage effect in vivo may be determined by a variety of factors including the tumor type and stage, the degree of macrophage infiltration and their functional polarization. Unfortunately, the in vivo mechanisms responsible for the anti-tumor activity of macrophages are still unclear. Current promising strategies to target tumor macrophages in vivo include pharmacological agents capable to re-polarize them towards a classically activated phenotype or to inhibit their suppressive properties. PMID- 18508452 TI - Neurotoxic effects of antineoplastic drugs: the lesson of pre-clinical studies. AB - Several antineoplastic drugs induce severe toxic damage of the peripheral nervous system and chemotherapy-induced peripheral neurotoxicity (CIPN) can be dose limiting. Moreover, CIPN signs and symptoms can be permanent and severely impair the patients' quality of life even after drug withdrawal. Despite extensive investigation, the exact mechanisms of neurotoxic action at the basis of CIPN are not completely known and it is likely that they can be at least in part different from the mechanisms of antineoplastic action of the drugs. A possible instrument to investigate on this important issue is represented by the evaluation of the effect of compounds used to reduce the toxicity of antineoplastic drugs in pre clinical and clinical settings. This review will be focused on the most clinically-relevant neurotoxic antineoplastic drugs and on the results obtained with several different classes of putative neuroprotectants. PMID- 18508453 TI - Platelets and wound healing. AB - Platelets help prevent blood loss at sites of vascular injury. To do this, they adhere, aggregate and form a procoagulant surface favoring thrombin generation and fibrin formation. In addition, platelets express and release substances that promote tissue repair and influence processes such as angiogenesis, inflammation and the immune response. They contain large secretable pools of biologically active proteins, while newly synthesized active metabolites are also released. Although anucleate, activated platelets possess a spliceosome and can synthesize tissue factor and interleukin-1beta. The binding of secreted proteins within a developing fibrin mesh or to the extracellular matrix can create chemotactic gradients favoring the recruitment of stem cells, stimulating cell migration and differentiation, and promoting repair. The therapeutic use of platelets in a fibrin clot has a positive influence in clinical situations requiring rapid healing. Dental implant surgery, orthopaedic surgery, muscle and tendon repair, skin ulcers, hole repair in eye surgery and cardiac surgery are situations where the use of autologous platelets accelerates healing. We now review the ways in which platelets participate in these processes. PMID- 18508454 TI - Endothelial connexins are down-regulated by atherogenic factors. AB - Connexins are single polypeptides that assemble to form paired connexon hexamers participating in gap-junctional intercellular communication. In addition, unpaired connexons at cell membrane also act as channels connecting cytosols and extracellular space. These channels' properties plus other unique functions of connexins give the molecules significant roles in endothelial cells, which mainly express connexin43 (Cx43), Cx40, and Cx37. In vitro studies have shown that expression of endothelial connexins are regulated by both physiological and pathological factors, a majority of which are involved in atherogenesis. In vascular disorders, endothelial connexins are differentially regulated. However, down-regulation of gap junctions is a common phenomenon. These findings suggest that reduced expression of endothelial gap junctions is a potential indicator of endothelial dysfunction and warrant investigators to explore the molecular mechanisms as well as therapeutic implications. PMID- 18508455 TI - Purification and characterization of dipeptidyl peptidase IV-like enzymes from bovine testes. AB - Until now, only recombinant forms of dipeptidyl peptidase (DPP) 8 and 9 have been characterized. We purified non DPPII-non DPPIV enzymes from a natural source. A first DPP8/9-like enzyme was enriched 1160-fold from bovine testes and identified as 'DPP9-like enzyme' by using an anti-DPP9 antibody. A second 576-fold enriched preparation ('DPP enriched peak 3') also showed DPP8/9-like activity. SDS-PAGE analysis showed that the DPP9-like enzyme had a monomeric molecular mass of approx. 100 kDa. Size exclusion chromatography generated a native molecular mass of 164 kDa for the DPP9-like enzyme and one of 234 kDa for the DPP enriched peak 3, suggesting that both proteins appeared to be dimeric. Both enriched preparations and rDPP8 showed roughly similar substrate specificity and inhibitor profiles. The DPP9-like enzyme and the DPP enriched peak 3 possessed a neutral pH optimum and were stable at -80 degrees C. We can conclude that the natural DPP9 like enzyme and the DPP enriched peak 3 are closely related to the recombinant forms of human DPP9 and DPP8. PMID- 18508456 TI - The central attentional limitation and executive control. AB - A central attentional limitation is assumed to be one reason why processing costs emerge in situations in which people do two things at once. This limitation causes that processes in two tasks are processed in serial order, if they require simultaneous access to the capacity-limited resource, which is called bottleneck interference. The present article links together recent knowledge about the psychological mechanisms and about the neural implementation of bottleneck interference. First, new findings are reviewed about the location of bottleneck interference in the processing chain, about its relation to the content of the processed information and its dependence on practice. In addition, further new evidence is reviewed that suggests that the bottleneck does not result from a passive occupation of the attention-limited resource by some process. Instead it is suggested that the serial order of processes at a bottleneck results from the involvement of control processes regulating the order of access to the capacity limited resource. Neuroimaging research suggests that these control processes are associated with activation in regions of the lateral prefrontal cortex, which can be dissociated from the neuro-anatomical implementation of other control functions during dual-task processing. PMID- 18508458 TI - Tremorolytic effects of adenosine A2A antagonists: implications for parkinsonism. AB - Drug-induced tremulous jaw movements in rats have been used as a model of parkinsonian tremor. Because adenosine A2A antagonists have antiparkinsonian effects, the present experiments were conducted to study the ability of adenosine A2A antagonism to reverse the tremulous jaw movements produced by the antipsychotic drugs pimozide, haloperidol and reserpine. In one group of studies, rats received daily injections of the dopamine antagonist pimozide, and on day 8 they received injections of pimozide plus various doses of the A2A antagonists KW 6002 or MSX-3. KW 6002 and MSX-3 suppressed pimozide-induced tremulous jaw movements, reduced catalepsy, and increased locomotion. MSX-3 also suppressed the jaw movements induced by haloperidol and reserpine. In addition, local injections of MSX-3 into the ventrolateral neostriatum suppressed pimozide-induced tremulous jaw movements. Thus, adenosine A2A antagonism can reverse the tremulous movements induced by antipsychotic drugs, which is consistent with the hypothesis that antagonism of adenosine A2A receptors can result in antiparkinsonian effects. Adenosine A2A antagonists may be useful for their tremorolytic effects, and may help in treating both idiopathic and antipsychotic-induced parkinsonian symptoms. PMID- 18508457 TI - The p38 MAPK stress pathway as a tumor suppressor or more? AB - p38 mitogen-activated protein kinases (p38 MAPKs) are a group of serine/threonine protein kinases that together with ERK (extracellular signal-regulated kinases) and JNK (c-Jun N-terminal kinases) MAPKs act to convert different extracellular signals into specific cellular responses through interacting with and phosphorylating downstream targets. In contrast to the mitogenic ERK pathway, mammalian p38 MAPK family proteins (alpha, beta, gamma, and delta), with and without JNK participation, predominantly regulate inflammatory and stress response. Recent emerging evidence suggests that the p38 stress MAPK pathway may function as a tumor suppressor through regulating Ras-dependent and -independent proliferation, transformation, invasion and cell death by isoform-specific mechanisms. A selective activation of a stress pathway to block tumorigenesis may be a novel strategy to control human malignancies. PMID- 18508459 TI - The kinetochore and spindle checkpoint in mammals. AB - The two daughter cells that result from each and every cell division receive an identical set of chromosomes. This is accomplished by pulling each copy of a pair of duplicated sister chromatids to opposite poles during mitosis. Inaccuracies in this process lead to aneuploidy, which is a major cause of birth defects and can facilitate the rise of malignancies. Such inaccuracies are prevented in normal cells by the mitotic checkpoint (also known as the spindle assembly checkpoint) that halts cell cycle progression in mitosis when as little as a single chromosome is not properly attached to the mitotic spindle. This review focuses on molecular aspects of mitotic checkpoint signaling in mammals, including sensing improper attachments and transducing this information to the cell-cycle machinery. PMID- 18508461 TI - The roles of chemokines in leukocyte recruitment and fibrosis in systemic sclerosis. AB - Systemic sclerosis (SSc, scleroderma) is an autoimmune disease characterized by excessive extracellular matrix deposition and vascular injury in the skin and other visceral organs. Although the pathogenesis remains unclear, interactions among leukocytes, endothelial cells, and fibroblasts are likely to be central to the pathogenesis of the disease. Chemokines mediate the leukocyte chemotaxis and migration through endothelia into the organ tissues, leading to the interaction between leukocytes and fibroblasts. While amounts of literatures reported chemokine abnormalities in SSc, which might explain the altered accumulation of effector leukocyte subsets in the affected tissues. Among various chemokines, monocyte chemoattractant protein-1 (MCP-1/CCL2) likely has the most critical role for tissue fibrosis in SSc. Although therapeutic effect for targeting MCP-1 has been demonstrated in mouse models of SSc or fibrotic disorders, it is unknown whether this strategy is effective in human clinical trials. Here recent data will be reviewed on the pathogenic role of chemokines and their receptors in SSc. PMID- 18508460 TI - Atherosclerosis as a disease of failed endogenous repair. AB - As coronary artery disease (CAD) continues to be the primary cause of mortality, a more in-depth understanding of pathophysiology and novel treatments are being sought. The past two decades have established inflammation as a driving force behind CAD--from endothelial dysfunction to heart failure. Recent advances in stem/progenitor cell biology have led to initial applications of progenitor cells in CAD continuum and have revealed that atherosclerosis is, at least in part, a disease of failed endogenous vascular repair. Several key progenitor cell populations including endothelial progenitor cells (AC133+/CD34+ population), vascular progenitors (CD31+/CD45(low) population), KDR+ cells and other bone marrow subtypes are mobilized for vascular repair. However, age and risk factors negatively impact these cells even prior to clinical CAD. Sex-based differences in progenitor cell capacity for repair have emerged as a new research focus that may offer mechanistic insights into clinical CAD discrepancies between men and women. Quantifying injury and cell-based repair and better defining their interactions should enable us to halt or even prevent CAD by enhancing the repair side of the repair/injury equation. PMID- 18508462 TI - Dipeptidyl peptidase IV (DPP IV) and related molecules in type 2 diabetes. AB - Dipeptidyl peptidase IV (DPP IV) is a widely distributed physiological enzyme that can be found solubilized in blood, or membrane-anchored in tissues. DPP IV and related dipeptidase enzymes cleave a wide range of physiological peptides and have been associated with several disease processes including Crohn's disease, chronic liver disease, osteoporosis, multiple sclerosis, eating disorders, rheumatoid arthritis, cancer, and of direct relevance to this review, type 2 diabetes. Here, we place particular emphasis on two peptide substrates of DPP IV with insulin-releasing and antidiabetic actions namely, glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). The rationale for inhibiting DPP IV activity in type 2 diabetes is that it decreases peptide cleavage and thereby enhances endogenous incretin hormone activity. A multitude of novel DPP IV inhibitor compounds have now been developed and tested. Here we examine the information available on DPP IV and related enzymes, review recent preclinical and clinical data for DPP IV inhibitors, and assess their clinical significance. PMID- 18508463 TI - MtDNA mutations, functional decline and turnover of mitochondria in aging. AB - Aging is a complex biological process that involves gradual function deterioration in various tissues and organs of an individual. Mitochondrial function decline can lead to cellular overproduction of reactive oxygen species (ROS) and increase in oxidative damage to biological molecules in the aging process. We have hypothesized that increased production of ROS by the mitochondria in affected tissues in patients with mitochondrial diseases and elderly subject results in increased oxidative stress and oxidative damage. Due to the similarity of human aging process to diseases related to bioenergetic function decline and mitochondrial DNA (mtDNA) alterations, aging is sometimes viewed as a "chronic" version of such diseases. Recent studies have also established that the expression profiles of several clusters of genes are altered, oxidative modification of proteins are increased and their turnover are decreased in tissues of old human subjects and animals. Accumulating evidence has suggested that mtDNA mutations, oxidative stress, defective disposal of dysfunctional proteins and a slower turnover of mitochondria are associated with aging. PMID- 18508464 TI - Perioperative cell-mediated immune response. AB - Innate and acquired immunity play a pivotal role in the host defense response. Pain, surgical stress, tissue injury and invasive micro-organisms are known to modulate complex immune responses in patients undergoing major surgery, which can lead to subsequent increased susceptibility to postoperative infections. Anesthetics may influence the immune response indirectly through modulation of the neurohumoral response or directly by acting on immune competent cells. In particular, cell-mediated immune balance seems to be affected by anesthetics and this might account for anesthetic-dependant risk of postoperative infections. Consequently, in order to fully understand the immunomodulating properties and ensuing clinical relevance of anesthetics it will be necessary to investigate each agent individually and in a variety of clinical settings. The existing research in this field, most of which is experimental, has yielded different results. The goal of further clinical studies must be to establish the immunomodulating properties of individual anesthetic agents so that selection can be tailored to the individual patient's pre-operative immune status and intraoperative course. PMID- 18508465 TI - Potassium channels in the basal ganglia: promising new targets for the treatment of Parkinson's disease. AB - A large number studies indicate that potassium (K+) channels play important roles in cellular signaling in both excitable and nonexcitable cells. Moreover, a considerable number of K+ channels within the nervous system appear to mediate diverse cellular signaling, including regulation of neurotransmitter release, neuronal excitability, and cell volume. Recent studies on the K+ channel gene expression in the basal ganglia reveal dysfunctions of various K+ channels (e.g., Kv, K(ATP), Kir2 and SKCa), which may be involved in the pathogenesis of Parkinson's disease (PD). This review aims to provide an overview of our current understanding of the molecular mechanisms involved in K+ channel functions in the basal ganglia, and an insight on how to exploit K+ channels as therapeutic targets in the treatment of PD. PMID- 18508466 TI - Inflammatory bowel diseases: multiple benefits from therapy with dipeptidyl- and alanyl-aminopeptidase inhibitors. AB - Inflammatory bowel diseases (IBD) are driven by imbalances in innate and acquired immune response. In IBD two dysregulated T cell subsets are in the focus of interest: activated effector T cells and regulatory T cells. These T cell subsets are characterized by a strong expression of the ectopeptidases dipeptidyl peptidase IV (DPIV /CD26) and aminopeptidase N (APN/CD13), which are thought to a role in the control of immune activation and in regulating cellular communication by hydrolyzing bioactive polypeptides. Since inhibitors of both enzymes were shown to be effective in limiting immune activation processes in vitro as well as in vivo, they emerged as new drug candidates for the treatment of diseases associated with an imbalanced T cell response, such as IBD. In this review we intent to throw light on the putative role of DPIV, APN and related enzymes in the regulation of immune and non-immune processes in inflammatory bowel diseases, on possible benefits from peptidase inhibitor therapy in these diseases as well on the gaps of knowledge in this field. PMID- 18508468 TI - Pharmacokinetic aspects of reduced nicotinamide adenine dinucleotide (NADH) in rats. AB - Nicotinamide adenine dinucleotide (NADH) plays a major role in cellular metabolism and mitochondrial dysfunction and is thought that NAD+/NADH decrease neuronal degeneration and improve behavioral deficits. This potential use of NAD+ or NADH as neuroprotective drugs requires an insight on the pharmacokinetic properties of these compounds. For this reason, we assessed the absorption of NADH in the small intestine in vitro using the everted gut sac technique. We show an enteral absorption of the intact NADH molecule. In the gut sac, NADH had a concentration-independent absorption rate of about 5 percent and the in vivo laser-induced fluorescence spectroscopy revealed a relatively quick absorption of NADH starting after a few minute reaching a plateau (about 5 percent ) after 20 30 minutes. Theses results show that, should NADH be protected against the acidic conditions of the stomach, NADH is absorbed principally in the small intestine. PMID- 18508467 TI - Molecular analysis of early host cell infection by Trypanosoma cruzi. AB - Trypanosoma cruzi, the causative agent of Chagas heart disease, infects heart and other cells leading to cardiac arrest frequently followed by death. The disease affects millions of individuals in the Americas and is posing health problems because of blood transmission in the US due to large Latin American immigration. Since the current drugs present serious side effects and do not cure the chronic infection, it is critically important to understand the early process of cellular infection at the molecular and structural levels to design novel inhibitors to block T. cruzi infection. In this review, the authors critically analyze the molecular and cellular basis of early T. cruzi infection and discuss the future directions in this area. The candidate T. cruzi invasive genes and host genes involved in the process of early infection are just beginning to be understood. The trypanosome invasive proteins are excellent targets for intervention. The progress made in the cell biology of T. cruzi infection will also facilitate the development of novel cell-based therapies to ameliorate the disease. PMID- 18508469 TI - Spatial integration and its moderation by attention and acetylcholine. AB - Attention is often regarded as a mechanism by which attended objects become perceptually more salient, akin to increasing their contrast. We demonstrate by means of human psychophysics that attention is better described as a mechanism that reduces contextual integration, thereby ensuring that task irrelevant information is prevented from influencing the processing of task relevant information. To investigate possible neuronal bases of this phenomenon we studied the effects of attention on spatial integration in V1 of the macaque monkey. In line with our psychophysical results, attention directed to parafoveal locations reduced spatial integration by reducing the summation area of V1 neurons. Additionally we measured length tuning in V1 in the presence and absence of externally applied acetylcholine in V1 of the marmoset monkey. The effects of acetylcholine application and attention were largely similar. Acetylcholine reduced spatial integration by reducing the neuron's summation area. These data demonstrate that attention can alter perceptual and neuronal spatial integration, and that acetylcholine might contribute to task dependent receptive field dynamics. PMID- 18508470 TI - The role of the multifunctional peptide LL-37 in host defense. AB - Neutrophil granules contain several antimicrobial peptides (AMPs) that are important effector molecules of innate immunity. In mammals, the main families of these peptides are the cathelicidins and defensins. Several defensins have been characterized in humans, while there is only one human cathelicidin, designated LL-37. This peptide is stored in specific granules of neutrophils in an inactive proform, which is processed extracellularly to the mature active peptide LL-37 and the propart cathelin after neutrophil degranulation. Apart from exhibiting a broad antimicrobial spectra, it is now evident that LL-37 possesses several additional functions that are related to host defense. Examples of such functions are chemotactic, endotoxin neutralizing, angiogenic and wound healing activities. These effects of LL-37 reveal a role as a mediator between innate and adaptive immunity. This review is giving an overview of the different immunological effects exerted by LL-37 and the physiological significance of these functions in immunity. PMID- 18508471 TI - Voltammetric investigation of surface-confined proteins. AB - Development of sensitive and selective methods to detect proteins at trace levels is of great biological importance. Via derivatization with a bifunctional cross linker 4-maleimidobutyric acid N-hydroxysuccinimide ester (GMBS) and an electrochemical marker 11-ferrocenyl-1-undecanethiol (Fc-SH), voltammetric determination of surface-confined proteins electrostatically adsorbed onto the polyelectrolyte of poly (sodium 4-styrenesulfonate) (PSS) or poly(allylamine hydrochloride) (PAH)-covered surfaces could be realized. The utilization of PSS or PAH was anticipated to reduce the nonspecific adsorption of the proteins on the surface. Two kinds of proteins with no redox activity or exhibiting complex or ill-defined voltammetric peak(s), i.e. the positively charged lysozyme and negatively charged metallothionein (MT) were demonstrated. Due to the incorporation of the bifunctional reagent GMBS and the redox active Fc groups onto the protein-modified electrodes, well-defined voltammetric peaks of high signal intensity were obtained. The anodic peak heights were found to be dependent on the surface density of the proteins electrostatically binded to the polyelectrolyte-coated surface. The present method can measure lysozyme concentration as low as 0.1 nM. PMID- 18508472 TI - Being a mouse in a man's world: what TMEV has taught us about human disease. AB - Choosing an appropriate animal model to study a disease is guided by a variety of factors including but not limited to the questions being asked, availability of reagents, knowledge of the animal species, personal biases of the researcher, and in some cases, cost and availability of facilities to effectively investigate the model. The validity of an animal model can be further complicated when the etiology of the disease is incompletely defined. Examples of these diseases include multiple sclerosis (MS) and type 1 diabetes (T1D). In addition to host genetics, epidemiological studies have implicated infectious agents, in particular viruses as triggers of these diseases. Thus many studies of these diseases have focused on modeling the interactions of viruses and the host immune response in vivo in small animals. Theiler's murine encephalomyelitis virus (TMEV) infection of mice has been used for over 30 years as a model of virus induced demyelination. TMEV induces a MS-like disease in susceptible strains of mice but does not cause pathology in humans. While some researchers may question the rationale for using a non-human pathogen to model human disease, the TMEV model of central nervous system (CNS) demyelination has permitted study of some aspects of human MS which would have been difficult to address in other models of the disease. Despite being 'merely a disease of mice,' many of the findings in the Theiler's virus model are directly applicable to the human condition, and studies from the model are responsible for our current understanding of mechanisms of pathology and clinical disability in human MS. In this review we will present some of the key findings from the TMEV model in the context of human disease. PMID- 18508473 TI - Apelin-induced vascular smooth muscle cell proliferation: the regulation of cyclin D1. AB - Apelin is the endogenous ligand of the G protein-coupled receptor, APJ. Vascular smooth muscle cells express both apelin and APJ, which are important regulatory factors in the cardiovascular and nervous systems. Importantly, APJ is also involved in the pathogenesis if HIV-1 infection. We investigated whether vascular smooth muscle cell proliferation was regulated through an apelin-pERK1/2-cyclin D1 signal transduction pathway. Apelin-13 significantly stimulated vascular smooth muscle cell proliferation and increased cell cycle progression. Apelin-13 a decreased the proportion of cell in the G0/G1 phase while increasing the number of cells in S phase. Apelin-13 also increased the levels of cyclin D1, cyclin E and pERK1/2. Treatment of cells with the MEK inhibitor PD98059 attenuated the apelin-3-induced pERK1/2 activation. Similarly, treatment with PD98059 partially diminished the apelin-13-induced expression of cyclin D1 and vascular smooth muscle cell proliferation. Taken together, these data established that apelin-13 stimulates vascular smooth muscle cell proliferation by promoting the G1-S phase transition, and that this effect is mediated in part by an apelin-pERK1/2-cyclin D1 signal cascade. PMID- 18508474 TI - Surgical brain injury: prevention is better than cure. AB - Neurosurgical procedures can cause inevitable brain damage resulting from the procedure itself. Unavoidable cortical and parenchymal incisions, intraoperative hemorrhage, brain lobe retraction and thermal injuries from electrocautery can cause brain injuries attributable exclusively to the neurosurgical operations and collectively referred to as surgical brain injury (SBI). This particular brain damage cannot be demarcated from the underlying brain pathology and has not been studied previously. Recently, we developed rat and mouse models to study SBI and the underlying cellular mechanisms. The animal modeling mimics a neurosurgical operation and causes commonly encountered postoperative complications such as brain edema following blood brain barrier (BBB) disruption, and neuronal cell death. Furthermore, the SBI animal model allows screening of known experimental neuroprotective agents and therapeutic agents being tried in clinical trials as possible pretreatments before neurosurgical procedures. In the present review, we elaborate on SBI and its clinical impact, the SBI animal models and their clinical relevance, and the importance of blanket neuroprotection before neurosurgical procedures. PMID- 18508475 TI - The Epstein-Barr virus (EBV) protein EB is an mRNA export factor essential for virus production. AB - The EBV early protein EB2 (aka Mta, SM and BMLF1) shares properties with mRNA export factors. It shuttles between the cytoplasm and the nucleus, and interacts with RNA both in vitro and in vivo but with no apparent sequence specificity. EB2 induces the cytoplasmic accumulation of mRNAs generated from intronless and intron-containing genes, likely through interactions with cellular export factors of the TAP/p15 pathway. Using a cell line carrying a viral genome with the EB2 gene deleted, it has been shown that EB2 is essential for the production of infectious virions by facilitating the nuclear export of a subset of early and late viral mRNAs, a function regulated by CK2 phosphorylation of EB2. There are docking sites for both CK2 subunits and for the heterotetrameric enzyme in the EB2 N- and C-terminal domains. Accordingly, EB2 and CK2 co-purify as a complex in which CK2 phosphorylates EB2. CK2 phosphorylation of EB2 at one of the Ser-55, Ser-56 and ser-57 is critical for its mRNA export function and as a consequence, for infectious virus production. PMID- 18508476 TI - Genotypic variation of Candida albicans during orthodontic therapy. AB - Candida is a common oral, opportunistic pathogen. The aim of this longitudinal study was to analyze the genotypes of sequential isolates of Candida albicans in a cohort of healthy consistent candidal carriers ('carriers') during fixed orthodontic appliance therapy. 11 of 97 subjects were 'carriers'. Candida isolates from baseline samples (T0) and sequential visits after insertion (T1 to T10) were speciated and RAPD fingerprint patterns of 101 sequential Candida albicans isolates were analyzed with PCR and gel electrophoresis. A similarity coefficient (SAB) for each pair of strains was calculated and clusters of similar strains grouped using Dendrogram analysis of the RAPD gel profiles. The composite dendrogram of all isolates indicated that the Candida populations in this cohort are genotypically dissimilar although collected from the same geographic locale and, from a similar healthy, age group. Genotypes of a majority of the cohort (6 of 11 consistent carriers) show minor evolutionary variations in profile and genotypic 'shuffling', implying this may be a survival mechanism of this common, human, opportunist pathogen residing in a hostile oral environment. PMID- 18508477 TI - Potassium channels in the basal ganglia: promising new targets for the treatment of Parkinson's disease. AB - A large number studies indicate that potassium (K+) channels play important roles in cellular signaling in both excitable and nonexcitable cells. Moreover, a considerable number of K+ channels within the nervous system appear to mediate diverse cellular signaling, including regulation of neurotransmitter release, neuronal excitability, and cell volume. Recent studies on the K+ channel gene expression in the basal ganglia reveal dysfunctions of various K+ channels (e.g., Kv, K(ATP), Kir2 and SKCa), which may be involved in the pathogenesis of Parkinson's disease (PD). This review aims to provide an overview of our current understanding of the molecular mechanisms involved in K+ channel functions in the basal ganglia, and an insight on how to exploit K+ channels as therapeutic targets in the treatment of PD. PMID- 18508478 TI - A ground truth based comparative study on clustering of gene expression data. AB - Given the variety of available clustering methods for gene expression data analysis, it is important to develop an appropriate and rigorous validation scheme to assess the performance and limitations of the most widely used clustering algorithms. In this paper, we present a ground truth based comparative study on the functionality, accuracy, and stability of five data clustering methods, namely hierarchical clustering, K-means clustering, self-organizing maps, standard finite normal mixture fitting, and a caBIG toolkit (VIsual Statistical Data Analyzer--VISDA), tested on sample clustering of seven published microarray gene expression datasets and one synthetic dataset. We examined the performance of these algorithms in both data-sufficient and data-insufficient cases using quantitative performance measures, including cluster number detection accuracy and mean and standard deviation of partition accuracy. The experimental results showed that VISDA, an interactive coarse-to-fine maximum likelihood fitting algorithm, is a solid performer on most of the datasets, while K-means clustering and self-organizing maps optimized by the mean squared compactness criterion generally produce more stable solutions than the other methods. PMID- 18508479 TI - Proteomic identification of novel proteins associated with Lewy bodies. AB - The manifestation of Lewy bodies (LB) in the brain is a hallmark of Parkinson's disease. Here, we present a comprehensive analysis of protein elements in Lewy bodies by comparative mass spectrometry. Cortical LB inclusions were enriched by sucrose gradient centrifugation from postmortem brains, and a negative control sample was prepared from specimen without LB pathology. Whereas approximately 550 proteins were identified in the LB-enriched sample by mass spectrometry, quantitative comparison with the control sample revealed that approximately 40 proteins were co-enriched with alpha-synuclein, the major component in Lewy bodies. As expected, the list of proteins included previously reported constituents, such as those involved in protein folding, membrane trafficking and oxidative stress. More interestingly, we discovered in the LB-enriched sample several kinases (MAPKK1/MEK1, protein kinase C, and doublecortin-like kinase), a novel deubiquitinating enzyme (otubain 1), and numerous ubiquitin ligases (KPC and SCF). The proteomic studies provide enzyme candidates to investigate the regulation of alpha-synuclein and/or other LB proteins, which may contribute to the formation of Lewy bodies and the toxicity of alpha-synuclein in the related neurodegenerative disorders. PMID- 18508480 TI - 4-Thiazolidinones: a novel class of hepatitis C virus NS5B polymerase inhibitors. AB - In a quest to identify novel compounds targeting HCV viral replicase, we evaluated a new series of 4-thiazolidinone derivatives (18 compounds). Our in vitro NS5B RdRp inhibition analysis with a series of 2',4'-difluoro-4 hydroxybiphenyl-3-carboxylic acid (2-(5-nitro-2-furyl/substituted phenyl)-4 thiazolidinone-3-yl) amides (1-7) yielded IC50 values ranging between 45-75 microM. Of these, lead compound 6: 2',4'-difluoro-4-hydroxybiphenyl-3-carboxylic acid(2-(2-fluorophenyl)-4-thiazolidinone-3-yl)amide exhibited an IC50 value of 48 microM and inhibited NS5B non-competitively with respect to UTP and exhibited a mixed mode of inhibition with respect to RNA. Molecular docking of thiazolidinone derivatives within the allosteric site of NS5B yielded significant correlation between their calculated binding affinity and IC50 values. Taken together, these data suggest that the 4-thiazolidinone scaffold may be optimized for generating new analogues with improved anti-NS5B potency. PMID- 18508481 TI - RNA processing control in avian retroviruses. AB - Upon integration into the host chromosome, retroviral gene expression requires transcription by the host RNA polymerase II, and viral messages are subject RNA processing events including 5'-end capping, pre-mRNA splicing, and polyadenylation. At a minimum, RNA splicing is required to generate the env mRNA, but viral replication requires substantial amounts of unspliced RNA to serve as mRNA and for incorporation into progeny virions as genomic RNA. Therefore, splicing has to be controlled to preserve the large unspliced RNA pool. Considering the current view that splicing and polyadenylation are coupled, the question arises as to how genome-length viral RNA is efficiently polyadenylated in the absence of splicing. Polyadenylation of many retroviral mRNAs is inefficient; in avian retroviruses, approximately 15 percent of viral transcripts extend into and are polyadenylated at downstream host genes, which often has profound biological consequences. Retroviruses have served as important models to study RNA processing and this review summarizes a body of work using avian retroviruses that has led to the discovery of novel RNA splicing and polyadenylation control mechanisms. PMID- 18508482 TI - Pathologic roles of prorenin and (pro)renin receptor in the eye. AB - Recent reports indicated that tissue renin-angiotensin system (RAS) was upregulated and angiotensin II type 1 receptor signaling plays crucial roles in ocular inflammation and neovascularization; however, the precise mechanism for activating tissue RAS had not been defined until recently. (Pro)renin receptor, a recently identified molecule existing in the major organs but not in the circulation, has attracted growing attention as an activator of tissue RAS. When the handle region of the prorenin prosegment binds to (pro)renin receptor, prorenin undergoes a conformational change to its enzymatically active state without the conventional proteolysis of the prorenin prosegment. Systemic treatment with a peptide with the structure of the handle region (handle region peptide; HRP), which competitively binds to (pro)renin receptor as a decoy peptide and inhibit the nonproteolytic activation of prorenin, resulted in the suppression of retinal inflammation and neovascularizaion in the rodent models. Retinal expression of RAS-related inflammatory and angiogenic molecules, such as intercellular adhesion molecule-1, monocyte chemotactic protein-1, and vascular endothelial growth factor, was also suppressed with application of HRP. These findings demonstrate that nonproteolytically activated prorenin plays a significant role in the ocular inflammation and neovascularization. PMID- 18508483 TI - Centromeric chromatin in fission yeast. AB - A fundamental requirement for life is the ability of cells to divide properly and to pass on to their daughters a full complement of genetic material. The centromere of the chromosome is essential for this process, as it provides the DNA sequences on which the kinetochore (the proteinaceous structure that links centromeric DNA to the spindle microtubules) assembles to allow segregation of the chromosomes during mitosis. It has long been recognized that kinetochore assembly is subject to epigenetic control, and deciphering how centromeres promote faithful chromosome segregation provides a fascinating intellectual challenge. This challenge is made more difficult by the scale and complexity of DNA sequences in metazoan centromeres, thus much research has focused on dissecting centromere function in the single celled eukaryotic yeasts. Interestingly, in spite of similarities in the genome size of budding and fission yeasts, they seem to have adopted some striking differences in their strategy for passing on their chromosomes. Budding yeast have "point" centromeres, where a 125 base sequence is sufficient for mitotic propagation, whereas fission yeast centromeres are more reminiscent of the large repetitive centromeres of metazoans. In addition, the centromeric heterochromatin which coats centromeric domains of fission yeast and metazoan centromeres and is critical for their function, is largely absent from budding yeast centromeres. This review focuses on the assembly and maintenance of centromeric chromatin in the fission yeast. PMID- 18508484 TI - Her2 cross talk and therapeutic resistance in breast cancer. AB - The HER2 receptor tyrosine kinase is amplified and/or overexpressed in approximately 30 percent of metastatic breast cancers. Interactions and cross signaling from the HER2 receptor to other growth factor receptors may potentially contribute to therapeutic resistance. In this review, we discuss HER2 receptor cross talk with the estrogen receptor and implications toward resistance to endocrine therapies. We also review mechanisms of resistance to the HER2-targeted antibody trastuzumab, including signaling from other members of the HER family, increased signaling through the PI3-kinase pathway, and cross talk from the insulin-like growth factor-I receptor to HER2. Finally, we will provide perspective on how HER2 receptor cross talk may provide critical information for developing novel therapeutic options for HER2-overexpressing breast cancers. PMID- 18508485 TI - Roles of MCP-1 in development of HIV-dementia. AB - The encephalopathy caused by HIV, known clinically as HIV-associated dementia (HAD) and pathologically as HIV encephalitis (HIVE), results from intense infiltration of mononuclear cells, productive replication of the virus in monocyte-derived macrophages/microglia, abortive replication in astrocytes and activation of macrophages/microglia and astrocytes leading to neuronal degeneration in the brains of infected persons. Recent findings have suggested that development of HAD is based more on the activation process than on direct evidence of virus replication in the brain. Since HAD is based on the encephalitic process, major studies have been directed to the mechanisms regulating the inflammatory process. Monocyte chemoattractant protein 1, MCP-1, is a chemokine that is implicated in this process and also in the development of activation in the brain. In this review, we have attempted to identify mechanisms that induce expression of MCP-1 in the brain and the role that it plays in recruitment of mononuclear cells from blood to brain and in the activation processes of inflammatory and neural cells that lead to development of degenerative changes in the neuronal population. PMID- 18508486 TI - Molecular dynamics simulations of Alzheimer Abeta40 elongation and lateral association. AB - Amyloid-beta (Abeta) peptides can elongate in the fibril axis and associate in the lateral direction. We present detailed atomic Abeta models with different in register intermolecular beta-sheet-beta-sheet associations. We probe structural stability, conformational dynamics, and association force of Abeta oligomers with various sizes and structures for both wild-type and mutated sequences using molecular dynamics (MD) simulations. MD simulations show that an Abeta oligomer that is laterally associated through the C-terminal-C-terminal interface is energetically more favorable than other oligomers with the N-terminal-N-terminal and C-terminal-N-terminal interfaces. We further develop a simple numerical model to describe the kinetics of Abeta aggregation process by considering fibril elongation and lateral association using a Monte Carlo algorithm. Kinetic data suggest that fibril elongation and lateral association are mutually competitive. Single-point mutations of Glu22 or Met35 at the interfaces have profound negative effects on intermolecular beta-sheet-beta-sheet association. These disease related mutants (E22K, E22Q, and M35O) display more flexible structures, weaker lateral association, and stronger elongation tendencies than the wild type, suggesting that amyloid oligomerization and neurotoxicity might be linked to fibril longitudinal growth. PMID- 18508487 TI - Renin and prorenin disappearance in humans post-nephrectomy: evidence for binding? AB - To study the distribution of kidney-derived renin-angiotensin system (RAS) components in humans, we monitored the decline in plasma prorenin, renin, angiotensin (Ang) I and Ang II post-nephrectomy. Prorenin and renin decreased biphasically, prorenin displaying a slower elimination. The distribution half life was similar for both. Angiotensins followed the disappearance of renin. One two days post-nephrectomy, stable plasma concentrations at 5-10% (renin and angiotensins) and 25-30% (prorenin) of pre-nephrectomy levels were reached. The total amount of kidney-derived renin and prorenin in the body was approximately 10 times as much as the amount in blood. Prorenin also originated at extrarenal sites. The renin levels in anephrics corresponded with the percentage of prorenin that in vitro has a so-called 'open conformation' (i.e., displays enzymatic activity), suggesting that renin in anephrics is in fact 'open' prorenin. Haemodialysis nor captopril significantly affected the level of any RAS component in anephrics. In conclusion, renal renin/prorenin enter tissue sites in humans, and renal renin is the main determinant of plasma angiotensins. Whether prorenin contributes to tissue angiotensin generation in humans remains to be determined. PMID- 18508488 TI - Mammary epithelial stem and progenitor cells and the prolactin pathway. AB - Prolactin is a pleiotropic peptide hormone and cytokine that is secreted from the pituitary gland and locally within various tissues of the body for autocrine and paracrine signal transduction. It controls proliferation and differentiation in a number of body tissues and increasing evidence indicates that it controls these functions in undifferentiated stem and progenitor cells of adult tissues, such as mesenchymal stem cells, hematopoietic progenitors, neural stem cells, oligodendrocyte precursor cells and possibly in mammary gland stem/progenitor cells. These roles in these undifferentiated cell types also implicate prolactin in the stem cell theory of cancer, supporting its known roles in cancer formation and progression. PMID- 18508489 TI - In silico study on the effect of F19T mutation on amyloid-beta peptide (10-35). AB - The wild type (WT) amyloid-beta (10-35) peptide, Abeta (10-35), and its F19T mutant have been studied by molecular dynamics simulations at 340 K in explicit water solvent each for over 3.4 ms. The WT peptide has a strong preference to form an E22-K28 loop (44% of total conformations) and a reasonable stability for a strand-loop-strand (SLS, L17-M35) (9%). The F19T mutant has a significantly lower population of E22-K28 loop (14%) and SLS structure (1.7%), but has a high population of Q15-D23 loop (48%). A specific interaction pattern among D23, V24, E22 and K28 was found to stabilize the E22-K28 loop in WT. Our results are in agreement with several experimental observations including: (1) the NOE constraints for the Abeta are reproduced; (2) the regions (15-23) and (22-28) can form loops; (3) the WT peptide is more structured than the F19T mutant. The current results also support our early proposal that the SLS structure might be important intermediate for monomer deposition to fibril, which explains the experimental fact that F19T mutant resists deposition to fibril. PMID- 18508490 TI - The role of chemokines in glomerulonephritis. AB - Leukocyte infiltration to glomeruli plays an essential role in the pathogenesis of glomerulonephritis. Pathophysiological roles of chemokines and their cognate receptors have shed light on the detailed molecular mechanisms of leukocyte trafficking and activation both in clinical and experimental settings of glomerulonephritis. Infiltrating leukocytes and glomerular resident cells interact to promote and exacerbate glomerular injury, eventually leading to glomerulosclerosis. Further, recent studies on chemokines have expanded their universe beyond leukocyte migration to glomeruli, to include homeostasis, development and protection of resident cells in glomeruli. New insights into proteinuria have been uncovered by the regulation of chemokine system. The intervention of chemokines and their cognate receptors may have therapeutic potential to slow the progression of glomerulonephritis. PMID- 18508491 TI - Microenvironmental regulation of E-cadherin-mediated adherens junctions. AB - The interaction between tumor cells and the microenvironment has substantial effects on tumor cell behavior by influencing cell-cell as well as cell-matrix contacts. The underlying molecular mechanisms are only partially unraveled. In this review we focus on the influence of the stromal microenvironment, especially collagen type I and type III on cellular adhesion and epithelial to mesenchymal transition (EMT). Extensive studies have emphasized that components of the microenvironment such as fibrillar collagen or growth factors like transforming growth factor beta are involved in induction of dedifferentiation of epithelial cells accompanied by disruption of the E-cadherin adhesion complex and reduced E cadherin concentrations. On the molecular level many different proteins have been identified which are involved in the regulation of EMT, such as activation of integrins, intracellular kinases such as Src, focal adhesion kinase (FAK) or phosphatidylinositol-3 kinase (PI3-kinase) and alteration of catenin phosphorylation. The reduced cellular adhesion influences the tissue integrity and allows tumor cells to disseminate from the primary tumor representing an early step in cancer metastasis. PMID- 18508492 TI - Roles of CD4+CD25(high) FOXP3+ Tregs in lymphomas and tumors are complex. AB - CD4+CD25(high)FOXP3+ regulatory T cells (Tregs) play an important role in the maintenance of immunological self-tolerance by suppressing autoimmune responses and anti-tumor immune responses. The current model suggests that epithelial tumor cells recruit Tregs to inhibit anti-tumor immunity in the tumor microenvironment, which thus limits the efficiency of anti-tumor immune responses and immunotherapy. However, recent findings on Tregs in lymphomas have complicated this working model. The biopsy specimens of some lymphomas have significantly higher percentages of Tregs than that in tumor-free lymph nodes and normal peripheral mononuclear cells. Higher Tregs numbers in these lymphomas predict improved survival and prognosis of patients. In this brief review, we summarize the progress in following topics: (1) Tregs; (2) Tregs and T cell co-stimulation; (3) Tregs in lymphomas; and (4) Tregs in other Tumors. Further characterization of Tregs in lymphomas and other tumors will provide insight on the differential regulation of Tregs' function and survival, and define the potentials of Tregs based immunotherapeutics. PMID- 18508493 TI - Iron oxide-loaded liposomes for MR imaging. AB - In this study a liposome cell labeling system was developed for non-target specific labeling of glioma cells with superparamagnetic iron oxide nanoparticles for magnetic resonance imaging (MRI). A high non-target-specific uptake is ideal for in vitro labeling of cells and subsequently for cell tracking and visualization of phagocytic cells in vivo. The preparation of iron oxide-loaded liposomes was optimized and the biological properties of the liposomes were investigated. Cytotoxicity and cell viability were examined and showed limited cytotoxic effects. Non-target-specific labeling of glioma cells in vitro for subsequent specific labeling of molecules for MR imaging was tested by T2* weighted MRI at 3T. The glioma cells showed a strong initial uptake of the iron oxide liposomes and the uptake was not saturable within 24 h exposure. The uptake of liposomes was superior to non-coated magnetite nanoparticles. Using PEG-ylated liposomes, the non-specific uptake could be decreased fundamentally (86% lower) in comparison to conventional liposomes. Furthermore, the ability of liposomes as contrast agents for MR imaging was investigated. Cells labeled with iron oxide nanoparticles by treatment with liposomes showed a negative contrast in MRI and consequently successful cellular labeling. Thus, iron oxide-loaded liposomes are well suited for non-target-specific cell labeling for MR imaging. PMID- 18508494 TI - Anthrax-associated shock. AB - Recent events have brought attention to the potential of Bacillus anthracis as an agent of bioterrorism. The shock like state of anthrax is invariably associated with high mortality, despite anti-microbial and supportive therapy. Multi-system dysfunction is typical, including: enhanced vascular permeability, hemorrhage and inflammation. Important questions concerning the pathophysiology of anthrax associated shock remain unanswered, including the effects of B. anthracis infection on cardiac function. This review discusses the current state of knowledge regarding the pathophysiology of anthrax-associated shock. PMID- 18508495 TI - Effects of n-3 fatty acids on autoimmunity and osteoporosis. AB - Decreased consumption of n-3 fatty acids (FA) and diets rich in animal proteins, saturated fats and n-6 vegetable oils are associated with a higher incidence of cardiovascular disease (CVD), certain malignancies and autoimmune disorders such as rheumatoid arthritis and Systemic Lupus Erythematosus (SLE), and renal disease. Recent studies show that reduced calorie intake and supplementation of diet with n-3 FA delays the onset of autoimmune renal disease, primarily, due to increased antioxidant enzyme activities, decreased NF-kappaB activation and decreased IL-1, IL-6 and TNF-alpha mRNA expression in the kidney tissue. Studies in rodents show that addition of n-3 FA and soy protein to diet affords protection against bone loss induced by ovariectomy in mice due to NF-kappaB expression and decreased activation of osteoclasts. Together, the available evidence show that increased daily intake of dietary n-3 FA decreases the severity of autoimmune disorders, lessens the chance of developing CVD, and protects against bone loss during post-menopause. PMID- 18508496 TI - Roles of chemokines in renal ischemia/reperfusion injury. AB - Ischemia-reperfusion injury (IR) is a common and an important clinical cause of renal disease, such as renal transplantation, renal artery stenosis and following shock from any cause. Inflammatory reaction after IR is regulated by various kinds of mediators. Chemokines are major mediators of the inflammation, and regulate pro-inflammatory cytokine and adhesion molecule expression, and leukocyte infiltration and activation. Chemokines are the key players of inflammation, angiogenesis and fibrosis. These inflammatory processes mediated by chemokines were observed in not only experimental animal models, but also in human renal diseases with ischemic injury. A number of challenges of chemokine targeted therapy is trying to prevent the ischemic injury, and will give some beneficial effect on the injury. PMID- 18508497 TI - The possible biological role of metallothionein in apoptosis. AB - Metallothioneins (MTs) are a family of low molecular weight proteins with a high affinity for divalent metals. Metallothionein has been shown to have a protective role in apoptosis. Specifically, it controls the cellular zinc ion levels. The proper intracellular Zn2+ level maintains the fragmentation of DNA associated with caspase-3 activity. In cancer nests, MT has been identified in the peripheral regions and associated with higher cell proliferation rates and fewer positive apoptotic cells. The expression of MT in the stroma has been linked with tumor spread. The tumor stroma may respond to tumor growth and aggressiveness by the expression of MT, thus protecting itself from and developing a resistance to apoptosis. Multistep carcinogenesis is related to a breakdown in immune response that enables tumor progression and dissemination. This breakdown is a crucial for tumor survival. The evaluation of MT expression in cancer and its stroma seems to correlate with the level of immune system inhibition in cancer-adjacent stroma. PMID- 18508498 TI - Towards modeling of amyloid fibril structures. AB - Amyloid fibrils are associated with a number of debilitating diseases, including Alzheimer's disease and variant Creutzfeldt-Jakob disease. The elucidation of the structure of amyloid fibrils is an important step toward understanding the mechanism of amyloid formation and developing therapeutic agents for amyloid diseases. Despite great interests and substantial efforts from various research communities, deriving high-resolution structures of amyloid fibrils remains a challenging problem, due to the insolubility and non-crystalline nature of the fibrils. An array of experimental methods, such as electron microscopy, fiber diffraction, hydrogen-deuterium exchange, solid-state NMR, electron paramagnetic resonance spectroscopy and biochemical approaches, have been explored to study the problem, having yielded considerable amount of, though still partial, information about the fibril conformation. Computational modeling techniques can be used to predict and build structural models of amyloid fibrils, utilizing the available experimental data. Here, we describe a few computational methods for modeling of aggregate and fibril structures with a focus on protein threading based approaches and discuss the challenging issues ahead. PMID- 18508499 TI - Non-synaptic neuronal mechanisms of learning and memory in gastropod molluscs. AB - Gastropod molluscs provide important model systems for investigating the behavioral and neural basis of associative and non-associative learning. Habituation, sensitization, classical and operant conditioning are studied in motor reflex and central pattern generator circuits. Although synaptic plasticity has long been recognized as playing a key role in molluscan learning circuits, non-synaptic changes resulting in alterations in the excitability of neurons are increasingly recognized as an essential component of the memory trace. PMID- 18508500 TI - Is there a major role for adenosine A2A receptors in anxiety? AB - Clinical investigations, pharmacological studies and models of genetically modified rodents have implicated adenosine in the etiology and modulation of different types of anxiety. Caffeine, a non-selective adenosine antagonist, has been involved in many of them. Adenosine seems to interact with other neurotransmitter systems and with some substances like alcohol, which elevate the basal levels of adenosine. A growing body of data describes the role of adenosine A1 and A2A receptors on anxiety. However, a differential role of adenosine receptors is not very clear. A1 receptor antagonists seem to be anxiogenic, but the absence of any effect of some of them and the opposite effects of others does not strongly support this conclusion. Human studies suggest that there is a susceptibility locus for panic disorder and agoraphobia within the receptor A2A gene. On the other hand, pharmacological data do not advocate for a clear implication of the A2A receptor. More research in this area is needed. PMID- 18508501 TI - Potential role of adenosine A2A receptors in the treatment of schizophrenia. AB - Schizophrenia is a complex neuropsychiatric disorder characterized by cognitive deficits, and positive and negative symptoms. All antipsychotics currently used in clinical practice are dopamine D2 receptor antagonists. The idea that adenosine A2A receptor agonists might be of interest for the treatment of schizophrenia derived from studies showing the existence of antagonistic intramembrane interaction between A2A and D2 receptors. Based on results obtained in animal models, a putative antipsychotic-like profile of A2A agonists was put forward. However, A2A agonists were shown to have detrimental effects in animal models of learning and memory. Moreover, these compounds produce many peripheral side-effects which limits their use in clinical trials. On the other hand, The results concerning the influence of A2A receptor antagonists in animal models used in schizophrenia studies such as locomotor activity and prepulse inhibition are fairly controversial. Some cognitive enhancing properties of A2A receptor antagonists have also been found in rats. Recent results showing the existence of heteromeric A2A/D3 and A2A/mGlu5 receptor complexes seem to open up new perspectives on the search for novel therapies of schizophrenia. PMID- 18508502 TI - The GTPase Ran: regulation of cell life and potential roles in cell transformation. AB - The GTPase Ran plays a crucial role in nucleo-cytoplasmic transport of tumor suppressors, proto-oncogenes, signaling molecules and transcription factors. It also plays direct roles in mitosis, through which it regulates faithful chromosome segregation and hence the generation of genetically stable cells. Ran operates through a group of effector proteins. In this review we summarize growing evidence suggesting that deregulated activity of Ran or its effectors can contribute to pathways of cell transformation and facilitate tumor progression. PMID- 18508503 TI - Therapeutic implications of the nuclear factor-kappaB/nuclear receptor cross talk. AB - More and more evidence reveals that the transcription factor NF-kappaB plays a critical role in tumor development and progression and that it may constitute the missing link between inflammation and cancer. It turned out that many of the well known cancer drugs exert their anti-tumoral effect at least in part through modulating the activity of NF-kappaB. The potential of nuclear receptors to modulate the activity of this widespread transcription factor has repeatedly been reported and illustrates their enormous therapeutic potential. However, the efficacy of these liganded receptors is overshadowed by the occurrence of unwanted effects owing to their broad range of actions. Accordingly, researchers pursue the ambition to improve the specificity of nuclear receptor modulators. In this review we have explored the molecular mechanisms by which nuclear receptors interfere with NF-kappaB signalling and quoted the therapeutic implications of their cross-coupling. Strategies that are explored at the moment and that may hold great potential for the future are extensively reviewed. PMID- 18508504 TI - The roles of sPLA2-IIA (Pla2g2a) in cancer of the small and large intestine. AB - The mouse secretory phospholipase A2 group IIA (sPLA2-IIA) gene Pla2g2a has been identified as a susceptibility gene for cancer of the small and large intestine. Interestingly, unlike most previously identified tumor susceptibility genes, Pla2g2a does not behave like a classical oncogene or tumor suppressor gene. Hence, identification of its biological functions in tumor development may shed new light on general mechanisms that modulate colon cancer risk. So far, sPLA2 IIA has been proposed to play a role in anti-bacterial defense, inflammation and eicosanoid generation, in clearance of apoptotic cells, and in the Wnt signaling pathway. More recently, comparison of RNA expression profiles of colon from Pla2g2a-transgenic to Pla2g2a-deficient mice confirmed and even extended sPLA2 IIA's diverse biological effects. In this review we aim to summarize current knowledge about the various links of sPLA2-IIA to cancer of the gastro-intestinal tract, and propose several models to illustrate its putative biological effects on tumor development. PMID- 18508505 TI - Clinical manifestations of hereditary cystic kidney disease. AB - Genetic mutations of discrete loci are the cause of a diverse array of polycystic kidney disease syndromes which present in distinct, as well as overlapping, phenotypic and hereditary patterns. Since molecular diagnostics are not currently a feasible clinical tool for the diagnosis of most cystic kidney diseases, physicians must rely upon their clinical acumen and knowledge base in order to identify these patients. The goal of this manuscript is to review the hereditary patterns, basic epidemiology, and phenotypic features of the most common of the cystic renal diseases so as to increase the awareness of these renal diseases among practicing physicians. Specifically, the genetic and phenotypic features of autosomal dominant polycystic kidney disease, autosomal recessive polycystic kidney disease, nephronopthisis-medullary cystic kidney disease complex, Bardet Biedl syndrome, and oral-facial-digital syndrome type 1 will be reviewed. PMID- 18508506 TI - Nitric oxide the gatekeeper of endothelial vasomotor control. AB - The endothelium can elicit relaxations and contractions of the underlying smooth muscle cells. It does so by releasing vasodilator (EDRF) and vasoconstrictor (EDCF) mediators. Among the diffusible endothelial factors nitric oxide (NO) plays a key role, particularly in large blood vessels. This chapter briefly reviews the interactions between NO and the other vasomotor signals released by the endothelial cells. PMID- 18508507 TI - Transcription factors in autoimmune diseases. AB - Studies on the pathogenesis of autoimmune diseases have begun to delve into the molecular and cellular mechanisms, and transcription factors, as key regulators of immune effector cell development and function, have received growing attention. Their involvement has been investigated in immune cells, such as T and B lymphocytes, macrophages and neutrophils, but also end-organ tissues, such as synoviocytes, keratinocytes and epithelial cells, and has revealed particularly dominant roles for NF-kappaB, STAT and AP-1 family members. This review summarizes recent findings and current knowledge regarding the roles of transcription factors in autoimmunity, focusing on their role in pathogenesis, as evidenced by both biological and genetic studies, as well as the implications of these findings for anti-inflammatory therapies. PMID- 18508508 TI - Viral infections in the pathogenesis of autoimmune diseases: focus on type 1 diabetes. AB - Susceptibility to autoimmune diseases is dictated by a complex interplay of genetic determinants and environmental factors. Viral infections have long been suspected to be involved in the etiology of several autoimmune disorders. In particular, the incidence of type 1 diabetes worldwide is increasing at a yearly rate that cannot be solely attributed to genetic changes in the population and environmental factors certainly play an important role in the pathology of this autoimmune disease. In this review, we will focus our discussion on the evidence supporting a role for viral infections in the pathology of viral induced autoimmunity based on the example of type 1 diabetes. We will place a particular emphasis on the major candidate viruses and on the current state of knowledge regarding the mechanisms responsible for the induction of autoimmunity following viral infections. The lessons learned from type 1 diabetes provide a great framework of knowledge that can be applied to most viral-induced autoimmune diseases. PMID- 18508509 TI - The RSK factors of activating the Ras/MAPK signaling cascade. AB - The p90 ribosomal S6 kinase (RSK) constitute a family of serine/threonine kinases activated downstream of the Ras/mitogen-activated protein kinase (MAPK) pathway. In mammals, four RSK genes have been identified (RSK1, RSK2, RSK3 and RSK4), and RSK orthologues have also been described in D. melanogaster and C. elegans, but not in yeast or plants. The RSK isoforms are composed of two distinct and functional kinase domains that are activated in a sequential manner by a series of phosphorylation events. These enzymes were among the first substrates of extracellular signal-regulated kinase (ERK) to be discovered and have proven to be ubiquitous and multifunctional mediators of ERK signal transduction. While the RSK isoforms promote cell survival though the inactivation of several apoptotic effectors, they also appear to mediate cell growth and proliferation by simultaneously regulating substrates involved in gene transcription and mRNA translation. RSK1-4 are ubiquitously expressed in cell lines and tissues, and at present, little is known about specific and overlapping functions of individual RSK isoforms. The upregulation of RSK1 and RSK2 expression in different types of cancer suggest that they may be involved in oncogenesis and could potentially be targeted in anti-cancer therapies. The recent identification of specific RSK inhibitors will likely help addressing the biological functions of the RSK isoforms and their contributions in pathological conditions. PMID- 18508510 TI - Role of osteopontin in cellular signaling and metastatic phenotype. AB - Osteopontin (opn) is a glyco-phosphoprotein that is expressed and secreted by numerous human cancers. Opn has pivotal role in cell adhesion, chemotaxis, prevention of apoptosis, invasion, migration and anchorage-independent growth of tumor cells. Extensive research has demonstrated the pivotal role of opn in regulation of cellular signaling, which controls neoplastic and malignant transformation. The elevated expression of opn has been observed in a variety of cancers. Recently, substantial evidence has linked opn with tumor metastasis and poor prognosis. However, the understanding of the molecular mechanisms that define the role of opn in cell invasion and metastasis is incomplete. The following review will discuss the molecular structure of opn, its function role in tumor cell metastasis and its downstream target genes that activate invasive mechanisms. Understanding of the role of opn in neoplastic transformation and its cellular target genes may enable development of novel anti-cancer therapy approaches. PMID- 18508511 TI - Proteasome inhibitors: a therapeutic strategy for haematological malignancy. AB - The proteasome is a multicatalytic enzyme complex responsible for the regulated degradation of intracellular proteins. In recent years, inhibition of proteasome function has emerged as a novel anti-cancer therapy. Proteasome inhibition is now established as an effective treatment for relapsed and refractory multiple myeloma and offers great promise for the treatment of other haematological malignancies, when used in combination with conventional therapeutic agents. Bortezomib is the first proteasome inhibitor to be used clinically and a second generation of proteasome inhibitors with differential pharmacological properties are currently in early clinical trials. This review summarises the development of proteasome inhibitors as therapeutic agents and describes how novel assays for measuring proteasome activity and inhibition may help to further delineate the mechanisms of action of different proteasome inhibitors. This will allow for the optimized use of proteasome inhibitors in combination therapies and provide the opportunity to design more potent and therapeutically efficacious proteasome inhibitors. PMID- 18508512 TI - Grappling with the HOX network in hematopoiesis and leukemia. AB - The mammalian HOX gene network encodes a family of proteins which act as master regulators of developmental processes such as embryogenesis and hematopoiesis. The complex arrangement, regulation and co-factor association of HOX has been an area of intense research, particularly in cancer biology, for over a decade. The concept of redeployment of embryonic regulators in the neoplastic arena has received support from many quarters. Observations of altered HOX gene expression in various solid tumours and leukemia appear to support the thesis that 'oncology recapitulates ontogeny' but the identification of critical HOX subsets and their functional role in cancer onset and maintenance requires further investigation. The application of novel techniques and model systems will continue to enhance our understanding of the HOX network in the years to come. Better understanding of the intricacy of the complex as well as identification of functional pathways and direct targets of the encoded proteins will permit harnessing of this family of genes for clinical application. PMID- 18508513 TI - The heparanome and regulation of cell function: structures, functions and challenges. AB - The cell-extracellular matrix interface is a crowded space whose structure is dependent on macromolecular assemblies that are dynamic in time, molecular composition and location. Signals travel from one cell to another (or to the same cell) by the regulated assembly/disassembly of molecular complexes. These signals can evoke relatively simple biological responses such cell proliferation and migration, but once integrated, they guide cell fate in complex biological phenomena such as embryonic development and organism homeostasis. Heparan sulfate proteoglycans are ubiquitous components of this space and important actors of these processes in all tissue-organized life forms. A key feature of heparan sulfate is its size, 40 nm to 160 nm, which enables it to integrate self assembling macromolecular structures over substantial length scales. What is the structure of heparan sulfate? Why do we think heparan sulfate is so important? How do we try to explain its activity? What do we know about its interactions? These questions together with a final look to the future are the "menu" of this review. PMID- 18508514 TI - Neuropilins: a versatile partner of extracellular molecules that regulate development and disease. AB - Neuropilins are a vertebrate-specific family of membrane multidomain proteins. They are crucial for the embryonic development of neural and vascular systems, whereas in the adult organism they are implicated in many processes, such as angiogenesis and the immune response. Additionally, it has been shown that they are overexpressed in numerous types of tumours, which results in higher microvessel density and correlates with poor prognosis. Their functions have been linked to their binding partners: semaphorins/collapsins, vascular endothelial growth factors (VEGFs), fibroblast growth factors (FGFs), hepatocyte growth factor/scatter factor and heparin/heparan sulfate (HS). Multiplicity of ligands alongside complex formation with several membrane receptors makes neuropilins potential 'hub' proteins, which act as a scaffold for multimeric associations. This review focuses on the structural features of neuropilins that underpin their multiple molecular interactions and hence their function. PMID- 18508515 TI - Osteopontin as a target for cancer therapy. AB - Osteopontin (OPN) is a glycophosphoprotein cytokine that has multiple functions. OPN is expressed and secreted by various cells, and has a role in cell adhesion, chemotaxis, prevention of apoptosis, invasion, migration and anchorage independent growth of tumor cells. Extensive research has demonstrated the pivotal participation of OPN in the regulation of cell signaling which controls neoplastic and malignant transformation. The elevated expression of OPN has been observed in a variety of cancers. OPN has been linked with tumor metastasis and signifies a poor prognosis for the patient. This review details the mechanisms by which OPN facilitates these pathological events. It will also show that gaining an understanding of the mechanism of OPN's action at a cellular level has led to the development of a number of therapeutic strategies against the cytokine. These include inhibiting its expression, antagonizing cell surface receptor activation and blocking downstream cell signaling pathways. In addition to the potential of these therapies, serum levels of OPN could be used as a diagnostic and prognostic marker. The authors propose that with further research and development, osteopontin directed treatment could greatly enhance outcomes for cancer patients. PMID- 18508516 TI - Evolutionary constraints imposed by gene dosage balance. AB - The gene dosage balance hypothesis states that a concentration imbalance among components of macromolecules is often deleterious. Thus this notion potentially provides a mechanistic explanation for understanding genetic dominance and gene duplicability. Accumulating evidence emerged from recent genomic data has strongly supported this hypothesis. Further efforts are needed to understand dosage sensitivity in the context of organismal complexity and co-regulation of genes under dosage-balance constraints. PMID- 18508517 TI - Does the understanding of immune activation by RNA predict the design of safe siRNAs? AB - The innate immune system uses pattern recognition receptors (PRRs) such as Toll like receptors (TLRs) to recognize conserved pathogen-associated molecular patterns (PAMPs) expressed by microbes and to activate the initial phase of immune response. Both bacterial and viral nucleic acids activate TLRs resulting in cytokines and interferon production. Recent studies indicated that unmodified small interfering RNAs (siRNAs) corresponding to either mammalian (self) or microbial (non-self) RNA sequences can activate immunity through TLR7/8 in a sequence-dependent manner. Hence, the use siRNAs in humans will require understanding the mechanisms involved in the discrimination between self and non self RNAs. In the case where immunostimulation is not wanted, chemical modifications can prevent immune activation and reduce off-target effects, while preserving siRNA silencing potency. Interestingly, 2'-O-methyl modified RNAs not only evade immune sensing, but antagonize with immunostimulatory single- or double-stranded siRNAs to activate innate immunity. This review highlights the recent progress in understanding the siRNA sensing by innate immunity and presents a range of strategies allowing either the design of siRNAs with minimal or maximal immunostimulatory potency for therapeutic applications. PMID- 18508519 TI - CCL3 protects mice from corneal pathology during recurrent HSV-1 infection. AB - CCL2 and CCL3 are proinflammatory chemokines that are produced during the early stages of inflammation and are known to stimulate the migration of mononuclear cells to the site of inflammation,. Previous studies addressing the role of these chemokines during primary herpetic stromal keratitis (HSK), have suggested that CCL2 is involved in reducing corneal disease and that CCL3 is involved in promoting this disease. We addressed the role that these chemokines play in a recurrent model of HSK. Results from these studies did not demonstrate a significant role for CCL2 except for very early time points following reactivation of virus. Surprisingly, mice deficient in CCL3 did not have significantly reduced recurrent disease, , but in fact showed significantly enhanced disease. This argues that CCL3 might play an ameliorative role during recurrent HSK. In addition, we observed that these same CCL3 deficient mice showed increased resistance to viral-induced mortality following infection with HSV-1. Taken together, these results suggest that CCL3 plays a significant protective role during recurrent HSK and is involved in enhancing lethality. PMID- 18508518 TI - Mouse hepatitis virus infection of the CNS: a model for defense, disease, and repair. AB - Viral infection of the central nervous system (CNS) results in varied outcomes ranging from encephalitis, paralytic poliomyelitis or other serious consequences. One of the principal factors that directs the outcome of infection is the localized innate immune response, which is proceeded by the adaptive immune response against the invading viral pathogen. The role of the immune system is to contain and control the spread of virus within the CNS, and paradoxically, this response may also be pathological. Studies with a neurotropic murine coronavirus, mouse hepatitis virus (MHV) have provided important insights into how the immune system combats neuroinvasive viruses, and have identified molecular and cellular mechanisms contributing to chronic disease in persistently infected mice. PMID- 18508520 TI - Subcellular proteomics in neuroscience. AB - The brain is the most complex and dynamically organized organ of the human body, with a high degree of computation capability enabling the execution of a wide spectrum of physiological processes and behaviors. In the past decades a large number of genomics studies have been undertaken to investigate brain function and brain disorders, but despite these efforts many of the underlying molecular mechanisms still remain largely unknown. The implementation of mass spectrometry based quantitative proteomics in recent years enabled to tap into condition specific protein trafficking and protein interaction that are the key to organelle proteome (dys)function. The technology for neuroproteomics is still evolving; currently there are no standardized protocols. In this review we describe the most commonly used methods to prepare brain subcellular fractions suitable for proteomics analysis, and highlight the various approaches for quantitative neuroproteomics. PMID- 18508523 TI - Tandem repeats in human disorders: mechanisms and evolution. AB - One of the most compelling reasons for the study of repetitive DNA sequence in the human genome has been the instability of simple repeat sequences associating with a growing and an interesting group of disorders affecting the neurological, neuromuscular or developmental processes. As a result, the molecular processes that underlie this unique form of mutation and the pathological pathways that lead to the disorders are being uncovered rapidly and are being intensively investigated. Genes with expanded repeats exhibit either loss-of-function or gain of-function effect at the protein and/or RNA level. In this review, we aim to provide an overview of the recent advances in molecular pathology of disorders associated with heritable changes in the length of the repeat sequences, and examine how dynamism in these repeats is regulated. PMID- 18508521 TI - The diverse functions of Src family kinases in macrophages. AB - Macrophages are key components of the innate immune response. These cells possess a diverse repertoire of receptors that allow them to respond to a host of external stimuli including cytokines, chemokines, and pathogen-associated molecules. Signals resulting from these stimuli activate a number of macrophage functional responses such as adhesion, migration, phagocytosis, proliferation, survival, cytokine release and production of reactive oxygen and nitrogen species. The cytoplasmic tyrosine kinase Src and its family members (SFKs) have been implicated in many intracellular signaling pathways in macrophages, initiated by a diverse set of receptors ranging from integrins to Toll-like receptors. However, it has been difficult to implicate any given member of the family in any specific pathway. SFKs appear to have overlapping and complementary functions in many pathways. Perhaps the function of these enzymes is to modulate the overall intracellular signaling network in macrophages, rather than operating as exclusive signaling switches for defined pathways. In general, SFKs may function more like rheostats, influencing the amplitude of many pathways. PMID- 18508522 TI - Ciliary dysfunction in polycystic kidney disease: an emerging model with polarizing potential. AB - The majority of different cell types in the human body have a cilium, a thin rod like structure of uniquely arranged microtubules that are encapsulated by the surface plasma membrane. The cilium originates from a basal body, a mature centriole that has migrated and docked to the cell surface. The non-motile cilia are microtubule-based organelles that are generally considered sensory structures. The purpose of this review is to discuss the practicality of the ciliary hypothesis as a unifying concept for polycystic kidney disease and to review current literature in the field of cilium biology, as it relates to mechanosensation and planar cell polarity. The polycystins and fibrocystin localization at the cilium and other subcellular localizations are discussed, followed by a hypothetical model for the cilium's role in mechanosensing, planar cell polarity, and cystogenesis. PMID- 18508524 TI - Cdt1 and Geminin in cancer: markers or triggers of malignant transformation? AB - Cdt1 and its inhibitor Geminin are important regulators of replication licensing. In normal cells, a critical balance between these two proteins ensures that firing of each origin along the genome will take place only once per cell cycle. Cdt1 overexpression in cell lines and animals leads to aberrant replication, activates DNA damage checkpoints and predisposes for malignant transformation. Geminin inactivation mimics the effects of Cdt1 overexpression in cells and generates mitotic defects and abnormal chromosome segregation. Aberrant expression of Cdt1 and Geminin is thus linked to DNA replication defects, aneuploidy and genomic instability. These traits are considered integral to precancerous states and essential elements for malignant transformation. Moreover, Cdt1 and Geminin expression is deregulated in human tumor specimens and Cdt1 and Geminin may represent novel markers useful for cancer diagnosis and prognosis. PMID- 18508525 TI - Exploring energy landscapes of protein folding and aggregation. AB - Human diseases, such as Alzheimer's and Creutzfeldt-Jakob's are associated with misfolding and aggregation of specific proteins into amyloid fibrils sharing a generic cross-beta structure. The self-assembly process is complex, but once a nucleus is formed, rapid fibril formation occurs. Insight into the structures of the oligomers during the lag phase, varying between hours and days, is very difficult experimentally because these species are transient, and numerically using all-atom molecular dynamics because the time scale explored is on the order of 10-100 ns. It is therefore important to develop simplified protein models and alternative methods to sample more efficiently the conformational space. In the past few years, we have developed the activation-relaxation technique (ART nouveau) coupled to the OPEP coarse-grained force field. This review reports the application of ART-OPEP on protein folding and aggregation. PMID- 18508526 TI - Chondrogenesis of mesenchymal stem cells in gel-like biomaterials in vitro and in vivo. AB - Gel-like carrier materials were introduced into cell therapy of cartilage lesions to improve chondrocyte retention and distribution in the defect. Mesenchymal stem cells (MSC) are now discussed as an alternative cell source for repair. We here asked whether distinct gel-like carriers can support chondrogenesis of MSC in vitro and lead to stable cartilage-like transplants in vivo. Chondrogenesis of MSC embedded in collagen type I gel, fibrin glue, Matrigel and PuraMatrix peptide hydrogel was assessed and gene expression analysis, proteoglycan content, and collagen synthesis were quantified. Differentiated constructs were transplanted subcutaneously into SCID mice. All carriers supported chondrogenesis in vitro, but displayed material-dependent differences on COL2A1 gene expression, total collagen synthesis and proteoglycan deposition. The undesired calcification and microossicle formation in ectopic transplants in vivo was consistently suppressed by Matrigel. In sum, gel-like biomaterials were suitable carriers for MSC and promoted chondrogenesis. Suppression of calcification by particular gel-like materials makes their use even more attractive for MSC-based tissue engineering approaches in cartilage repair. PMID- 18508527 TI - Lymphocytic choriomeningitis infection of the central nervous system. AB - Viral infection of the central nervous system (CNS) can result in a multitude of responses including pathology, persistence or immune clearance. Lymphocytic choriomeningitis virus (LCMV) is a powerful model system to explore these potential outcomes of CNS infection due to the diversity of responses that can be achieved after viral inoculation. Several factors including tropism, timing, dose and variant of LCMV in combination with the development or suppression of the corresponding immune response dictates whether lethal meningitis, chronic infection or clearance of LCMV in the CNS will occur. Importantly, the functionality and positioning of the LCMV-specific CD8+ T cell response are critical in directing the subsequent outcome of CNS LCMV infection. Although a basic understanding of LCMV and immune interactions in the brain exists, the molecular machinery that shapes the balance between pathogenesis and clearance in the LCMV-infected CNS remains to be elucidated. This review covers the various outcomes of LCMV infection in the CNS and what is currently known about the impact of the virus itself versus the immune response in the development of disease or clearance. PMID- 18508530 TI - Production systems for recombinant antibodies. AB - Recombinant antibodies are the fastest growing class of therapeutic proteins. Furthermore, antibodies are key detection reagents in research and diagnostics. The increasing demand for antibodies with regards to amount and quality resulted in the development of a variety of recombinant production systems employing gram negative and gram-positive bacteria, yeast and filamentous fungi, insect cell lines as well as mammalian cell lines. More recently, antibodies were also successfully produced in transgenic plants and animals. Currently, the production of recombinant antibodies for therapy is performed in mammalian cell lines to reduce the risk of immunogenicity caused by non-human post-translational modifications, in particular glycosylation. However, novel strategies already allow human-like glycosylation patterns in yeast, insect cell lines and transgenic plants. Furthermore, therapeutic strategies not requiring glycosylation of the Fc portion have been conceived, most prominently using bispecific antibodies or scFv fusion proteins, which can be produced in bacteria. Here, we review all current antibody production systems considering their advantages and limitations with respect to intended applications. PMID- 18508528 TI - Regulation of neurite outgrowth by G(i/o) signaling pathways. AB - Neurogenesis is a long and winding journey. A neural progenitor cell migrates long distances, differentiates by forming a single axon and multiple dendrites, undergoes maturation, and ultimately survives. The initial formation of neurites during neuronal differentiation, commonly referred to as "neurite outgrowth," can be induced by a large repertoire of signals that stimulate an array of receptors and downstream signaling pathways. The G(i/o) family of heterotrimeric G-proteins are abundantly expressed in the brain and enriched at neuronal growth cones. Recent evidence has uncovered several G(i/o)-coupled receptors that induce neurite outgrowth and has begun to elucidate the underlying molecular mechanisms. Emerging data suggests that signals from several G(i/o)-coupled receptors converge at the transcription factor STAT3 to regulate neurite outgrowth and at Rac1 and Cdc42 to regulate cytoskeletal reorganization. Physiologically, signaling through G(i/o)-coupled cannabinoid receptors is critical for pro percentral nervous system development. As the mechanisms by which G(i/o)-coupled receptors regulate neurite outgrowth are clarified, it is becoming evident that modulating signals from G(i/o) and their receptors has great potential for the treatment of neurodegenerative diseases. PMID- 18508529 TI - Advances in molecular imaging of pancreatic beta cells. AB - The development of non-invasive imaging methods for early diagnosis of beta cell associated metabolic diseases, including type 1 and type 2 diabetes (T1D and T2D), has recently drawn interest from the molecular imaging community and clinical investigators. Due to the challenges imposed by the location of the pancreas, the sparsely dispersed beta cell population within the pancreas, and the poor understanding of the pathogenesis of the diseases, clinical diagnosis of beta cell abnormalities is still limited. Current diagnostic methods are invasive, often inaccurate, and usually performed post-onset of the disease. Advances in imaging techniques for probing beta cell mass and function are needed to address this critical health care problem. A variety of imaging techniques have been tested for the assessment of pancreatic beta cell islets. Here we discuss current advances in magnetic resonance imaging (MRI), bioluminescence imaging (BLI), and nuclear imaging for the study of beta cell diseases. Spurred by early successes in nuclear imaging techniques for beta cells, especially positron emission tomography (PET), the need for beta cell specific ligands has expanded. Progress for obtaining such ligands is presented. We report our preliminary efforts of developing such a peptidic ligand for PET imaging of pancreatic beta cells. PMID- 18508531 TI - Multi-faceted roles for CXC-chemokines in prostate cancer progression. AB - CXC-chemokines play an essential role in co-ordinating the function of the immune system. Increasingly, these small signaling molecules are recognized in facilitating communication between multiple cell types within the tumor microenvironment. This review will summarize the role of two members of this family, CXCL12 (stromal cell derived factor-1) and CXCL8 (interleukin-8) in promoting the disease progression of prostate cancer, the most prevalent non cutaneous cancer in men in western society and the second leading cause of death from cancer in men. Evidence for a role of these chemokines in underpinning the development and progression of this disease is supported by examination of prostate tissue and serum samples from prostate cancer patients, from biochemical and molecular investigations conducted on representative cell-based models of this disease and from observation of CXC-chemokine promoted growth and systemic dissemination of human prostate tumors in experimental in vivo models. The future potential of employing strategies to attenuate chemokine expression or alternatively to selectively block chemokine receptor signaling to effect greater long-term control or enhanced therapeutic response in this disease is also discussed. PMID- 18508532 TI - Gene expression profiling for the diagnosis and prognosis of acute myeloid leukaemia. AB - Acute myeloid leukaemia (AML) is a heterogeneous disease covering a range of morphological lineages and differentiation stages, but also has number of recurrent chromosomal abnormalities and mutations associated with prognosis. Because of the defined molecular and cytogenetic features, AML has been a focus of gene expression profiling studies and identified differentially expressed genes in the different diagnostic and cytogenetic sub-groups. These studies have lead to the development of expression based classifications and diagnostic chips with further studies aiding prognosis and therapeutic choices. PMID- 18508533 TI - Does MK5 reconcile classical and atypical MAP kinases? AB - MAP kinase-activated protein kinase 5 (MK5) was originally described as a protein kinase activated downstream of the p38 MAP kinase and is also named p38 regulated/activated protein kinase (PRAK). However, while MK5 is most similar in sequence to the two p38 regulated MAPKAP kinases MK2 and MK3, recent data has shown that in contrast to these enzymes MK5 is not activated in response to either cellular stress or pro-inflammatory cytokines. This lack of response to stimuli which cause robust activation of p38 MAP kinase in vivo is supported by data obtained using transgenic mice lacking MK5. Unlike animals lacking MK2 and MK3, MK5 null mice respond normally to endotoxic shock and display an unchanged pattern of cytokine expression in response to LPS. Clues as to the physiological function of MK5 have come from the recent observation that MK5 is uniquely regulated and activated following complex formation with the atypical MAP kinases ERK3 and ERK4. Thus, it is possible that MK5 is unique amongst the MAPKAP kinases in being regulated downstream of signaling pathways other than the classical MAP kinases p38 and ERK1/2. PMID- 18508534 TI - Cystatin protease inhibitors and immune functions. AB - Cystatins are natural tight-binding reversible inhibitors of cysteine proteases. They are wide spread in all living organisms (mammals, nematodes, arthropods etc.) and are involved in various biological processes where they regulate normal proteolysis and also take part in disease pathology. Many cystatins show changes in expression and/or localization, as well as changes in secretion, following certain stimuli acting on immune cells. In immune cells, cystatins interfere with antigen processing and presentation, phagocytosis, expression of cytokines and nitric oxide and these ways modify the immune response. Further, it has been suggested that cystatin-type molecules secreted from parasites down-modulate the host immune response. Precise understanding of the regulatory roles on proteolytic enzymes of endogenous and exogenous cystatins, such as those from parasites, will provide us with valuable insight into how immune response could be modulated to treat a specific disease. This review covers some specific functions of individual cystatins, with a particular focus on the relevance of cystatins to the immune response. PMID- 18508535 TI - The evolution of deep brain stimulation for neuropsychiatric disorders. AB - Deep brain stimulation (DBS) is the most focal method for stimulating the human brain. In contrast to lesions, DBS is nonablative, with the advantages of reversibility and adjustability. Thus, therapeutic effectiveness can be enhanced and stimulation-related side effects minimized during long-term patient management. While DBS is an approved adjunct therapy for severe, medication refractory movement disorders, it remains investigational in neuropsychiatry. However, experience to date, though limited, suggests that DBS may offer a degree of hope to patients with severe and treatment-resistant neuropsychiatric illness. Thus far, work in obsessive-compulsive disorder (OCD), the first psychiatric condition studied using modern DBS devices, has shown consistently positive results across multiple small-scale studies. Work in treatment-resistant Major Depressive Disorder (MDD) also suggests therapeutic potential in preliminary studies, generating cautious optimism for this indication. With the increase in potential applications, a number of clinical and preclinical research efforts have now focused on understanding the mechanisms of action of DBS. Further development of DBS for these and other illnesses with primarily behavioral symptoms will require thoughtful collaboration among multiple disciplines. PMID- 18508536 TI - Cytoskeletal thermal ratchets and cytoskeletal tensegrity: determinants of brain asymmetry and symmetry? AB - Evolution can be viewed as a dynamic process that leads to increased complexity. This process appears to be driven by the interplay between a breaking of symmetry in biological organisms that leads to increased differentiation and complexity on one hand, and the intrinsic tendency of physical systems to maintain symmetry on the other. Thus thermal ratchets act to break symmetry, suggesting they may have played an important role is the evolution of complexity, while physical systems, including biological ones, have a tendency to maintain symmetry. We propose that, in the brain, development is driven by a combination of asymmetry-creating properties of cytoskeletal thermal ratchets and by the symmetry-maintaining properties of cytoskeletal tensegrity architecture. PMID- 18508537 TI - Gene content of LUCA, the last universal common ancestor. AB - Comparative genomics and modern phylogenetic approaches allow us to infer the gene content of LUCA, the Last Universal Common Ancestor of all known currently living cellular organisms. Most of the estimates produce a putative LUCA with 500 1000 protein-coding genes and biochemically coherent metabolism, if the average rates of gene gains (gene emergence plus horizontal gene transfer) and gene losses per family are allowed to be close to each other. This estimate is not strongly sensitive to the topology of the Tree of Life, but the identity of the genes that are placed in LUCA may depend on the position of the deep branches and the root of the tree. PMID- 18508538 TI - The plasminogen activation system in inflammation. AB - Inflammation is an adaptive response to damage of vascularized tissues, which develops according to a stereotyped sequence governed by the local production of the so-called "chemical mediators of inflammation". Here we review the evidences indicating a role of the plasminogen activation system in the regulation of all the phases of the inflammation process. Plasminogen activation controls the formation of complement anaphylotoxins (responsible for vasodilatation, increase of venular permeability and leukocyte chemotaxis) and of bradykinin (which accounts for vasodilatation, increase of venular permeability and pain) by regulating the plasma contact system. The urokinase plasminogen activator and its cellular receptor, expressed on the surface of human leukocytes, provide a functional unit that, by regulating interaction of leukocytes with extracellular matrix, as well as its degradation, is critical for the migration of leukocytes and for their movement in the damaged tissues. By preventing excess fibrin accumulation in inflamed tissues, the plasminogen activation system also governs the proper evolution of the inflammatory exudates and prevents the possibility of a shift from acute to chronic inflammation. PMID- 18508539 TI - Genome evolution and speciation genetics of clawed frogs (Xenopus and Silurana). AB - Speciation of clawed frogs occurred through bifurcation and reticulation of evolutionary lineages, and resulted in extant species with different ploidy levels. Duplicate gene evolution and expression in these animals provides a unique perspective into the earliest genomic transformations after vertebrate whole genome duplication (WGD) and suggests that functional constraints are relaxed compared to before duplication but still consistently strong for millions of years following WGD. Additionally, extensive quantitative expression divergence between duplicate genes occurred after WGD. Diversification of clawed frogs was potentially catalyzed by transposition and divergent resolution- processes that occur through different genetic mechanisms but that have analogous implications for genome structure. How sex determination is maintained after genome duplication is fundamental to our understanding of why allopolyploidization is so prevalent in this group, and why clawed frogs violate Haldane's Rule for hybrid sterility. Future studies of expression subfunctionalization in polyploids will shed light on the role and purviews of cis- and trans-regulatory elements in gene regulation. PMID- 18508540 TI - Antisense DNA and RNA agents against picornaviruses. AB - Anti-picornaviral antisense agents are part of a broader group of nucleic acid based molecules developed for sequence-specific inhibition of translation and/or transcription of the target sequence through induced nuclease activity or physical hindrance. Three types of nucleic acid-based gene silencing molecules can be distinguished, including DNA-base antisense oligonucleotides (ASO), nucleic acid enzymes (ribozyme and DNAzyme) and double-stranded small interfering RNA (siRNA or microRNA). These antisense DNA and RNA molecules have been widely studied for gene functional studies and therapeutic purposes. In this review, we focus on drug development using ASO and siRNA strategies to inhibit picornavirus infections. The picornavirus genome organization and life cycle is described, followed by discussion of design considerations, chemical modifications and drug delivery approaches. Recent studies using antisense against picornavirus are reviewed. Finally, we compare the advantages and disadvantages of the antisense agents with those of other therapeutics, taking into consideration their limitations which need to be overcome to achieve the final goal of clinical application. PMID- 18508541 TI - Bone morphogenetic protein-7 (BMP7) in chronic kidney disease. AB - Bone morphogenetic protein-7 (BMP7) is a member of the BMP-subfamily of perhaps a dozen proteins within the TGFbeta-superfamily of cysteine-knot fold cytokine growth factors. BMP7 has pivotal functions during renal and eye development. In adult organisms, BMP7 is heavily expressed in kidney, specifically in podocytes, distal tubules and collecting ducts. The activity of BMP7 is reduced by inhibitors including some members of the dan-cerberus group and CTGF but can be enhanced by endoglin and KCP. Renal BMP7 disappears early in fibrogenic renal diseases which may facilitate progression. Exogenous administration of rhBMP7 or transgenic overexpression reduces renal fibrogenesis and apoptosis as well as transdifferentiation of epithelial cells. BMP7 improves maintenance of nephron function and structural integrity. These antifibrogenic activities result from inhibition of the nuclear translocation of TGFbeta-activated smad3 by smad6 downstream of BMP7-activated smad5. Although at present the beneficial effects of BMP7 have only been studied in rodent models of chronic renal diseases, there is promise for therapeutic utility of rhBMP7 or small molecule BMP7 agonists in patients. PMID- 18508542 TI - Trimeric G protein-dependent signaling by Frizzled receptors in animal development. AB - Receptors of the Frizzled family transduce important signals during animal development and are conserved from sponges to humans. Frizzled receptors belong to the superfamily of G protein-coupled receptors (GPCRs), but until recently were considered G protein-independent in their signaling. In the present article we review the extensive knowledge demonstrating the functions of trimeric G proteins in Frizzled signal transduction in vertebrates and lower animals. Other structural and functional similarities of Frizzled receptors and the GPCRs are also discussed. PMID- 18508543 TI - DNA detection using a radio frequency biosensor with gold nanoparticles. AB - This study presents a novel method for DNA detection with multi-layer AuNPs to enhance overall detection sensitivity. This essay achieves not only an innovative radio-frequency biosensor but also a critical signal amplification methodology. Results show that bandwidth change for multi-layer AuNP with hybridization of DNA exceeds that for the double-layer AuNP up to 0.5 GHz. Furthermore, the developed biosensor detection limit for the DNA set employed in this essay is currently 10 pM. A single base-pair mutation of the wild-type target DNA could be distinguished from the perfect match target DNA at the melting temperature of 47 degrees C with a temperature controlling system. Experimental results in this study indicate that the proposed biosensor and the developed amplification methodology are successful. As health care becomes much more essential in modern life, this biosensor has potential applications in a screening kit for recognizing, sensing, and quantifying biomolecules in real samples. PMID- 18508544 TI - Muscular response and adaptation to diabetes mellitus. AB - Diabetes mellitus (DM) is an epidemic medical challenge that threatens the health and life quality of people worldwide. DM impairs metabolic, neural and vascular function and thus has profound impacts on different systems and organs in the body. Though continuous endeavour has been made to study its etiology and mechanisms, no cure for DM has yet been found. DM development may be multi factorial. The skeletal muscle is one of the most important systems, involved in the development of DM, and affected by insulin. DM induces diverse functional, metabolic, and structural changes in the skeletal muscle. DM reduces the functional capacity of skeletal muscle leading to muscle weakness, causes metabolic disturbance characterized by reduced cellular glucose uptake and fatty acid oxidation, and structural changes with muscle atrophy, augmented lipid deposition, decreased mitochondria as well as muscle fiber transformation. DM induced changes in the skeletal muscle seem to be dependent on types and severity of DM as well as on muscle fibers. The central mechanism underlying these changes is impaired insulin action in the skeletal muscle. PMID- 18508545 TI - Probing protein aggregation using discrete molecular dynamics. AB - Understanding the role of biomolecular dynamics in cellular processes leading to human diseases and the ability to rationally manipulate these processes is of fundamental importance in scientific research. The last decade has witnessed significant progress in probing biophysical behavior of proteins. However, we are still limited in understanding how changes in protein dynamics and inter-protein interactions occurring in short length- and time-scales lead to aberrations in their biological function. Bridging this gap in biology probed using computer simulations marks a challenging frontier in computational biology. Here we examine hypothesis-driven simplified protein models in conjunction with discrete molecular dynamics in the study of protein aggregation, implicated in series of neurodegenerative diseases, such as Alzheimer's and Huntington's diseases. Discrete molecular dynamics simulations of simplified protein models have emerged as a powerful methodology with its ability to bridge the gap in time and length scales from protein dynamics to aggregation, and provide an indispensable tool for probing protein aggregation. PMID- 18508546 TI - The heterogeneity of diabetic neuropathy. AB - Diabetic neuropathy and its underlying pathogenesis are reviewed. It has been documented for some time that diabetic neuropathy differs in both human and experimental type 1 versus type 2 diabetes. Such differences are accounted for by impaired insulin action and signal transduction in type 1 diabetes, whereas hyperglycemia per se contributes equally to neuropathy in the two types of diabetes. Such differences in basic initiating factors and pathogenesis translate into differences in the functional and structural expressions of neuropathy in type 1 and type 2 diabetes. Type 1 neuropathy shows a more rapid progression with more severe functional and structural changes. Several experimental mono therapies have been tested over the last decades which unfortunately have not been efficacious. Therefore discrepancies in underlying pathogenetic mechanisms in the two types of diabetic neuropathy will have to be taken into account in the design of future therapies, which should target several key pathogenetic mechanisms. Therapies that meet these criteria include replacement of acetyl-L carnitine and replenishment of C-peptide in type 1 diabetic neuropathy. PMID- 18508547 TI - Role of Toll like receptor-activated dendritic cells in the development of autoimmunity. AB - The recognition of microbial stimuli by Toll-like receptors (TLRs) expressed on dendritic cells (DCs) is essential for the regulation of immune responses. DC activation via TLRs leads to the production of proinflammatory cytokines, chemokines and surface molecules that play a key role in the regulation and control of inflammatory reactions and adaptive immunity. Minor imbalances in the feedback control of TLR-activated innate immune cells have been associated with autoimmunity in genetically prone individuals. We review here recent studies indicating how TLR-mediated activation of innate immune cells, including DCs, may be involved in the development and/or maintenance of autoimmune responses in the presence of both endogenous and exogenous ligands. PMID- 18508548 TI - Immune regulation by invariant NKT cells in autoimmunity. AB - Invariant NKT cells are important regulators of T cell immunity and autoimmunity. In this review we describe evidence that supports their regulatory role in the prevention of autoimmune disease. Moreover, we will discuss the current knowledge on iNKT cell biology, antigen recognition, acquisition of a specific cytokine profile, and mechanism of action that suggest a key role for iNKT cells as negative regulators of autoimmune diseases. PMID- 18508549 TI - Recent acquisitions on the genetic basis of autoimmune disease. AB - In this review we will discuss recent progress from studies on the genetic basis of autoimmune disease and how this has advanced our understanding of the processes behind disease susceptibility and pathogenesis. We review the genetic associations with autoimmune and inflammatory disease discovered in the latest genome-wide association (GWA) scans, and discuss the importance of animal models both for generating candidates and for mechanistic studies. Investigating the natural variants of key immune regulatory molecules can give us an additional level of insight into their function and physiological regulation over gene knockouts. New data showing the association of multiple genes involved in pathogen defense highlights the potential role of infection in autoimmunity, and a more complete understanding of the pathways defective in genetically susceptible individuals will also give us a handle on how environmental and epigenetic factors may be impacting disease. PMID- 18508550 TI - B lymphocytes--chief players and therapeutic targets in autoimmune diseases. AB - For some time, B cells have been considered as passive actors, exclusively depending on T cell conductors that provide them with instructions to engage in antibody secretion. With further investigation, however, it became evident that B cells can exert a number of antibody-independent functions, capturing and concentrating antigen for presentation, producing cytokines, influencing T cell and dendritic cell responses, contributing distinct functions during the immune response, affecting lymphoid tissue structures, and, even participating in tissue repair. Because of their multiples functions, B cells are currently recognized to play a key role in a variety of antibody-, and T cell-mediated autoimmune diseases, including lupus, rheumatoid arthritis, type-1 diabetes and multiple sclerosis. This recent insight led to novel immuno-intervention strategies that target B cells, with beneficial effects in patients. While such novel therapeutic bio-drugs are being introduced into the clinical arena, research intensifies in order to identify novel targets and strategies whose ultimate goal is to knock out specifically pathogenic B cells, and to amplify the numbers and the activity of cells endowed with regulatory functions. PMID- 18508552 TI - The role of programmed-1 ribosomal frameshifting in coronavirus propagation. AB - Coronaviruses have the potential to cause significant economic, agricultural and health problems. The severe acute respiratory syndrome (SARS) associated coronavirus outbreak in late 2002, early 2003 called attention to the potential damage that coronaviruses could cause in the human population. The ensuing research has enlightened many to the molecular biology of coronaviruses. A programmed -1 ribosomal frameshift is required by coronaviruses for the production of the RNA dependent RNA polymerase which in turn is essential for viral replication. The frameshifting signal encoded in the viral genome has additional features that are not essential for frameshifting. Elucidation of the differences between coronavirus frameshift signals and signals from other viruses may help our understanding of these features. Here we summarize current knowledge and add additional insight regarding the function of the programmed -1 ribosomal frameshift signal in the coronavirus lifecycle. PMID- 18508551 TI - The role of chemokines during herpes simplex virus-1 infection. AB - Herpes simplex virus-type 1 is among the most prevalent and successful humans pathogens. Although infection is largely uncomplicated in the immunocompetent human host, HSV-1 infection can cause blinding corneal disease, and individuals with defects in innate or adaptive immunity are susceptible to herpes simplex encephalitis. Chemokines regulate leukocyte trafficking to inflamed tissues and play a crucial role in orchestrating the immune response to HSV-1 infection. In this review we will focus on the pathways that induce chemokine expression during HSV-1 infection and the implications of chemokine signaling on control of viral replication. PMID- 18508553 TI - Methods and protocols for the assessment of protein allergenicity and cross reactivity. AB - Allergy is a prevalent health problem in developed countries. With advances in genomic and proteomic technologies, there is a rapid increase in allergy-related data, including allergen sequences, allergic cross-reactivity, molecular structures, clinical measurements, and atmospheric concentrations. The more and more complex allergy data is fueling the need for advanced ways in information management and analysis. Computational methods and resources are increasingly the driving force in allergy research. For example, allergen-specific databases are important data sources for allergen characterization. T-cell and B-cell epitope prediction tools focus on identifying immunogenic regions on allergenic proteins. Allergenicity and cross-reactivity prediction tools are increasingly being applied to assess the potential allergenicity of proteins. This review provides an introduction to the growing literature in this area, with particular emphasis on recent developments in bioinformatics relevant to the study of allergens. PMID- 18508554 TI - Selectivity and invariance for visual object perception. AB - The sight of an object triggers a complex set of processes in the brain. Although it is already well established that object perception is performed by a hierarchical network, the so-called ventral visual pathway, we are only starting to understand how neurons along this pathway encode visual information at each processing stage. In this review, we discuss basic principles of neural coding for object perception and describe evidence showing that it mainly relies on two principles: selectivity and invariance. PMID- 18508555 TI - Test systems for measuring ocular parameters and visual function in mice. AB - New techniques are described to measure refractive state, pupil responses, corneal curvature, ocular dimensions and spatial vision in mice. These variables are important for studies on myopia development in mice, but they are also valuable for phenotyping mouse mutants and for pharmacological studies. PMID- 18508556 TI - Influenza vaccines. AB - Influenza viruses pose a major challenge to those concerned with global public health. Not only do influenza viruses cause yearly epidemics that are associated with slight changes in viral antigenicity, but occasionally new viruses cross from animal reservoirs into humans causing major pandemics. The most effective method to lower the mortality and morbidity associated with influenza is vaccination. In this review current and pending influenza vaccine technologies will be discussed in the context of both epidemic and pandemic influenza. PMID- 18508558 TI - Arterial remodeling in vascular disease: a key role for hyaluronan and versican. AB - Hyaluronan and versican are extracellular matrix (ECM) macromolecules that are present in low amounts in normal blood vessels, but increase dramatically in vascular disease. These ECM components are particularly enriched in intimal hyperplasia as seen in human restenotic lesions following balloon angioplasty and provide a permissive environment for arterial smooth muscle cell (ASMC) proliferation, migration, and macrophage adhesion. Interference with the association of hyaluronan and versican with the surface of ASMCs, either through short oliogosaccharides of hyaluronan or blocking antibodies to the hyaluronan receptor, CD44, blocks the proliferative and migratory response of these cells to growth factors, such as platelet derived growth factor (PDGF). Agents that interfere with the proliferative response of ASMCs and that are used in the treatment of restenosis, such as rapamycin, inhibit the synthesis of hyaluronan by these cells. Inhibition of versican by versican antisense blocks proliferation of SMCs. The synthesis of hyaluronan and versican is highly regulated and influenced by pro-inflammatory growth factors such as PDGF and transforming growth factor-beta (TGF-beta). PMID- 18508557 TI - Protein tyrosine phosphatases in the JAK/STAT pathway. AB - The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway is crucial in controlling cellular activities in response to extracellular cytokines. Dysfunctions of the JAK/STAT pathway result in various hematopoietic and immune disorders. The central events in regulating this pathway are tyrosine phosphorylation and dephosphorylation of the signaling components, which are carried out by protein tyrosine kinases and protein tyrosine phosphatases (PTP), respectively. Here, we review recent advances in the regulatory roles of PTPs, in particular, SHP2 phosphatase, in the JAK/STAT signaling pathway. PMID- 18508559 TI - The SH3 domain--a family of versatile peptide- and protein-recognition module. AB - Src homology 3 (SH3) domains were initially characterized as a prevalent protein module that recognizes proline-rich sequences, in particular those containing a PxxP motif. Recent studies have shown that the specificity and cellular function of SH3 domains are far more diverse than previously appreciated. Despite lacking distinguishing features, the ligand-binding surface of an SH3 domain can be molded to accommodate a variety of peptide ligands. Moreover, certain SH3 domains are capable of using surfaces distinct from the canonical ligand-binding site to engage a peptide or protein. The identification of novel motifs and domains recognized by the SH3 domain greatly expands the ligand pool and cellular function for this family. However, this also imposes the question as to how the specificity of the hundreds of human SH3 domains is regulated in a cell to ensure their proper functions. Here we review literature on the specificity of SH3 domains, with an emphasis on the structural basis of ligand recognition, and discuss mechanisms employed by SH3 domain-containing proteins to execute defined cellular functions through highly regulated SH3-ligand interactions. PMID- 18508560 TI - Immunostimulatory virotherapy using recombinant Sendai virus as a new cancer therapeutic regimen. AB - The utility of recombinant Sendai virus (rSeV) has been considerably examined over the last decade as a potent gene transfer candidate in a cytoplasmic gene expression system. Such risks as excessive immune responses associated with this virus administration in vivo however have limited its applicability in clinical settings as is the case with other viral vectors including adenoviruses. In consequence of extensive assessment on the mechanisms of immune responses against SeV, we found that ex vivo infection of immature dendritic cells (DCs) with SeV demonstrates their spontaneous maturation and activation. We applied this result to create a unique, representative, and powerful agent to activate DCs, namely rSeV-modified DCs (rSeV/DCs), for use in cancer immunotherapy. Use of this system in vivo resulted in the induction of efficient antitumor immunity against vascularized rodent tumors, including melanoma, hepatocellular carcinoma, neuroblastoma, squamous cell carcinoma, and prostatic cancer, and it even frequently associated with elimination of those tumors. These results indicate that rSeV could be a powerful immune booster for DC-based cancer immunotherapy that is worth investigating further. We propose a conceptual term "immunostimulatory virotherapy" to describe this new method of cancer therapy using the rSeV/DCs system. PMID- 18508561 TI - Knock-out mouse models of proprotein convertases: unique functions or redundancy? AB - The members of the proprotein convertase family play a central role in the processing and/or activation of various protein precursors involved in many physiological processes and various pathologies. The proteolysis of these precursors that occur at basic residues within the general motif (K/R)-(X)-(K/R) is mediated by the proprotein convertases PC1/3, PC2, Furin, PACE4, PC4, PC5 and PC7, whereas the proteolysis of precursors within hydrophobic residues performed by the convertase S1P/SKI-1 and the convertase NARC-1/PCSK9 seems to prefer cleavages at the motif LVFAQSIP. Here we provide a comprehensive overview of their remarkable complex roles as revealed by disruption of their genes individually using generalized or conditional approaches. PMID- 18508562 TI - Cochlear molecules and hereditary deafness. AB - Remarkable progress has been made in the past decade in identifying genes involved with deafness in man and mouse. The identification of these genes and functional analysis of the proteins they encode are paving the way towards a better understanding of the physiology and pathophysiology of the auditory system. Given the complexity of auditory transduction and diversity of cochlear structures, it is not surprising that an estimate of at least 1 percent of human protein-coding genes are involved in perception of sound. Over 400 distinct syndromes of which hearing loss is a component have been reported (www.ncbi.nlm.nih.gov/omim). Approximately 113 loci for monogenic disorders for which hearing loss is the only manifestation and therefore is nonsyndromic, have been mapped to the human genome (http:webhost.ua.ac.be/hhh/). As of August 2007, there are approximately 46 genes identified from these loci. Here, we review some of the major advances in our knowledge of auditory function within an evolving understanding of the structure and regulation of the machinery of hearing. PMID- 18508563 TI - Smad7 as a therapeutic agent for chronic kidney diseases. AB - Increasing evidence shows that transforming growth factor-beta TGF-beta1 (TGF beta1) is upregulated and plays a diverse role in renal fibrosis by stimulating extracellular matrix (ECM) production, while inhibiting renal inflammation. Recent studies have identified that TGF-beta1, once activated, signals through its downstream signaling pathway to exert its biological effects. It is now well accepted that TGF-beta regulates fibrosis positively by receptor-associated Smads including Smad2 and Smad3, but negatively by an inhibitory Smad, called Smad7. We and other investigators have shown that gene transfer of Smad7 is able to inhibit renal fibrosis in a number of experimental models of chronic kidney diseases, including obstructive nephropathy, remnant kidney disease, and autoimmune crescentic glomerulonephritis. Blockade of Smad2/3 activation is a major mechanism by which overexpression of Smad7 inhibits renal scarring. Furthermore, our recent findings also demonstrate that Smad7 plays a critical role in anti inflammation in chronic kidney diseases by blocking the NF.kappaB-dependent inflammatory pathway. Thus, Smad7 has a unique role in both anti-renal fibrosis and inflammation. These findings also indicate that targeting the TGF-beta/Smad signaling pathway by overexpressing Smad7 may provide a novel, specific, and effective therapy for chronic kidney diseases. PMID- 18508564 TI - Novel aspects of the renin-angiotensin-aldosterone-system. AB - The renin-angiotensin-aldosterone system (RAAS) play a pivotal role in the progression of renal disease. The RAAS has become much more complex in recent years with the identification of novel peptides that exhibit biological activity. There are novel pathways of angiotensin II (ANG II) generation independent of angiotensin converting enzyme (ACE). ANG II bind to at least two different receptors and prorenin/renin also exerts pathophysiological effects through binding to specific receptor. ANG II itself has emerged as a multifunctional cytokine exhibiting many non-hemodynamic properties such as acting as a growth factor and profibrogenic and proinflammatory cytokine. These profibrogenic and proinflammatory effects are mediated by other factors such as transforming growth factor beta (TGF-beta) and chemoattractants that are induced in the kidney by ANG II. Increased aldosterone levels contribute to renal injury, independent of blood pressure or ANG II. Numerous experimental and clinical studies have shown that ACE-inhibitors as well as AT1-receptor antagonists can prevent glomerulosclerosis and tubulointerstitial fibrosis. This review will highlight some of these novel insights into the RAAS in regards to renal injury. PMID- 18508565 TI - Temporal regulation of adenovirus major late alternative RNA splicing. AB - Adenovirus makes extensive use of alternative RNA splicing to produce a complex set of spliced mRNAs during replication. The accumulation of viral mRNAs is subjected to a temporal regulation, a mechanism that ensures that proteins that are needed at certain stages of the virus life cycle are produced in a timely fashion. The complex interactions between the virus and the host cell RNA splicing machinery has been studied in detail during the last decade. These studies have resulted in the characterization of two viral proteins, E4-ORF4 and L4-33K, that adenovirus uses to remodel the host cell RNA splicing machinery. Here I will review the current knowledge of how mRNA expression from the adenovirus major late transcription unit is controlled with a particular emphasis on how cis-acting sequence element, trans-acting factors and mechanisms regulating adenovirus major late L1 alternative RNA splicing is controlled. PMID- 18508566 TI - The multiple checkpoint functions of CHK1 and CHK2 in maintenance of genome stability. AB - Cell cycle checkpoints are pivotal mechanisms safeguarding genome stability. Cells that harbor defects in checkpoints are predisposed to genome instability and neoplastic transformation. Two structurally-unrelated protein kinases, CHK1 and CHK2, are implicated in several major checkpoints of the cell cycle, providing a crucial linkage between the upstream sensors of the checkpoints and the cell cycle engine. Variations of the ATM/ATR-CHK1/CHK2-CDC25-CDK axis underlie the molecular basis of the replication checkpoint, the intra-S phase checkpoint, and the G2 DNA damage checkpoint. Although some aspects of the pathway remain contentious, the ATM/ATR-CHK1/CHK2-p53-p21CIP1/WAF1-CDK axis is believed to play an important role in the G1 DNA damage checkpoint. Recent data also reveal that CHK1 may play a role in the spindle-assembly checkpoint. Finally, CHK1 and CHK2 are implicated in linking the cell cycle to diverse processes such as senescence and the circadian cycle. In this review article, we provide an overview of how the multi-tasking nature of CHK1 and CHK2 is achieved in vertebrate cells. PMID- 18508568 TI - EpCAM-targeted induction of apoptosis. AB - EpCAM is a well-established pancarcinoma-associated target antigen that has been used in a variety of therapeutic approaches. Of particular appeal are those strategies that aim to retarget and locally activate immune effector mechanisms involving apoptosis. Cancer cells typically employ various strategies to evade recognition and elimination by immune effector cells, including low or absent expression of MHCI molecules and active elimination of tumor infiltrating immune cells. In addition, cancer cells show an increased resistance towards endogenous pro-apoptotic stimuli due to aberrancies in their apoptotic machinery. However, compelling evidence indicates that cancer cells are often reliant on these molecular aberrations for continued cell survival. This pivotal role of immune evasion and apoptosis resistance has fueled the quest for therapeutic strategies that can selectively retarget and reactivate immune effector cells or molecules, whereby the balance of cellular fate of cancer cells is selectively tipped towards apoptosis. Here we review and discuss the perspectives for EpCAM-targeted apoptosis induction in cancer by EpCAM-selective bispecific antibodies and TRAIL fusion proteins. PMID- 18508567 TI - Mitochondrial mechanisms of sepsis-induced organ failure. AB - Sepsis is the leading cause of death in medical intensive care units. Though progress has been made in the early treatment of sepsis associated with hemodynamic collapse (septic shock), little is known about the pathogenesis of delayed organ dysfunction during sepsis. A growing body of data indicates that sepsis is associated with acute changes in cell metabolism, and that mitochondria are particularly susceptible. The severity of mitochondrial pathology varies according to host and pathogen factors, and appears to correlate with loss of organ dysfunction. In this regard, low levels of cell apoptosis and mitochondrial turnover are normally observed in all metabolically active tissues; however, these homeostatic mechanisms are frequently overwhelmed during sepsis and contribute to cell and tissue pathology. Thus, a better understanding of the mechanisms regulating mitochondrial damage and repair during severe sepsis may provide new treatment options and better outcomes for this deadly disease (30-60% mortality). Herein, we present compelling evidence linking mitochondrial apoptosis pathways to sepsis-induced cell and organ failure and discuss the implications in terms of future sepsis research. PMID- 18508569 TI - EpCAM in morphogenesis. AB - Embryonic development is one of the most complex biological phenomena that involves the appropriate expression and synchronized interactions of a plethora of proteins, including cell adhesion molecules (CAMs). Many members of the diverse family of CAMs have been shown to be critically involved in the correct execution of embryonic development. The Epithelial Cell Adhesion Molecule (EpCAM) is an atypical cell adhesion molecule originally identified as a marker for carcinoma. However, recent insights have revealed that EpCAM participates in not only cell adhesion, but also in proliferation, migration and differentiation of cells. All of these processes are known to be fundamental for morphogenesis. Here, we review the current literature that establishes EpCAM as a protein involved in morphogenesis, starting from the earliest stages of embryogenesis and ending in organogenesis. In addition, we provide directions for further elucidation of the role of EpCAM in embryogenesis. PMID- 18508570 TI - Oxidative stress in vascular senescence: lessons from successfully aging species. AB - Cardiovascular disease is a main cause of morbidity and a leading cause of death of elderly Americans. Studies identifying the pathophysiological mechanisms underlying cardiovascular aging hold promise to develop treatments to delay/prevent coronary artery disease and stroke in the elderly. Evidence supporting the roles of oxidative stress and inflammation in the cardiovascular aging process is presented in detail in this review. Mammalian lifespan ranges hundred-fold and we propose that long-living species may be useful models for successful cardiovascular aging in humans. Comparative studies exploiting the large differences in maximum lifespan potential and cardiovascular aging patterns may be particularly relevant. Comparisons of mechanisms related to oxidative stress, oxidative stress resistance and redox signaling between long-living species and shorter-living ones may elucidate key mechanisms for delaying cardiovascular aging. We discuss the potential use of three long-lived but mouse sized mammalian species, the naked mole-rat (Heterocephalus glaber), the white footed mouse (Peromyscus leucopus) and the little brown bat (Myotis lucifugus) to test predictions of the oxidative stress theory of aging and elucidate mechanisms by which cardiovascular aging can be delayed. PMID- 18508571 TI - The genetics of malignant melanoma. AB - Melanoma probably is the most aggressive cancer in humans and remains one of the leading causes of cancer death in developed countries. This review summarizes the most important alterations in protooncogenes and tumor suppressor genes that contribute to the pathogenesis of malignant melanoma, with a special emphasis on the involved signaling pathways. Our knowledge of the molecular biology of melanoma has been benefited from recent advances on high-throughput technologies analyzing wide genomic and gene expression profiles that have uncovered unknown candidate genes. To test the interactions between distinct pathways and of those with the environment a wealth of genetically modified animal models has been generated over the past years. Other studies have focused on the isolation of melanoma stem cells and on the characterization of signaling pathways that contribute to their survival and maintenance. A consequence of all these studies is the emergence of potential new strategies that could improve the still inadequate arsenal of therapeutic tools to fight against this fatal disease. PMID- 18508572 TI - TNF: a master switch for inflammation to cancer. AB - Chronic inflammation has long been associated with the development of cancer, ever since Rudolf Virchow's first observation that leukocytes were present in neoplastic tissue more than 130 years ago. Recent evidences have reignited the interest of cancer researchers in the exciting concept of an association between chronic inflammation and cancer. Tumor necrosis factor alpha (TNF-alpha), initially discovered as a result of its antitumor activity, has now been shown to be one of the major mediators of inflammation. Induced by a wide range of pathogenic stimuli, TNF-alpha induces other inflammatory mediators and proteases that orchestrate inflammatory responses. TNF-alpha is also produced by tumors and can act as an endogenous tumor promoter. The role of TNF-alpha has been linked to all steps involved in tumorigenesis, including cellular transformation, promotion, survival, proliferation, invasion, angiogenesis, and metastasis. How TNF-alpha acts as a masterswitch in establishing an intricate link between inflammation and cancer is the focus of this review. PMID- 18508573 TI - New insights in drug development for the non-small cell lung cancer therapy. AB - Non-small cell lung cancer (NSCLC) remains a major problem worldwide. Since most patients with NSCLC have advanced disease at diagnosis, to date, chemotherapy, with third-generation platinum-based doublets, represents the standard of care. However, a plateau has been reached with the use of cytotoxic chemotherapy in advanced NSCLC. Advances in the knowledge of tumour biology and mechanisms of oncogenesis have granted the singling out of several molecular targets for NSCLC treatment. To date, erlotinib and gefitinib, epidermal growth factor receptor tyrosine kinase (EGFR-TK) inhibitors have been licensed, erlotinib worldwide and gefitinib in Asian countries, for refractory NSCLC. Currently, bevacizumab, an anti-vascular endothelial growth factor (VEGF) monoclonal antibody, is the only clinically available antiangiogenic agent licensed, in combination with carboplatin plus paclitaxel, for first-line therapy of advanced NSCLC patients in the United States. Several new biologic agents are being evaluated in clinical research and some of them, such as ZD6474, sorafenib and sunitinib, due to the reported preliminary results and the oral administration seem to be promising targeted agents for the treatment of NSCLC. Aim of this review is to discuss about the new insights in targeted agents development for the treatment of NSCLC patients. PMID- 18508575 TI - The neuroprotective mechanism of action of the multimodal drug ladostigil. AB - The recent therapeutic approach in which drug candidates are designed to possess diverse pharmacological properties and act on multiple targets has stimulated the development of the multimodal drug, ladostigil (TV3326) ((N-propargyl-(3R) aminoindan-5yl)-ethyl methyl carbamate). Ladostigil combines neuroprotective effects with monoamine oxidase -A and -B and cholinesterase inhibitory activities in a single molecule, as a potential treatment for Alzheimer's disease (AD) and Lewy Body disease. Preclinical studies show that ladostigil has antidepressant and anti-AD activities and the clinical development is planned for these dementias. In this review, we discuss the multimodal effects of ladostigil in terms of neuroprotective molecular mechanism in vivo and in vitro, which include the amyloid precursor protein processing; activation of protein kinase C and mitogen-activated protein kinase pathways; regulation of the Bcl-2 family members; inhibition of cell death markers and up-regulation of neurotrophic factors. Altogether, these scientific findings make ladostigil a potentially valuable drug for the treatment of AD. PMID- 18508574 TI - Converging roles for sphingolipids and cell stress in the progression of neuro AIDS. AB - Sphingolipids are a class of lipids enriched in the central nervous system that have important roles in signal transduction. Recent advances in our understanding of how sphingolipids are involved in the control of life and death signaling have uncovered roles for these lipids in the neuropathogenesis of HIV-associated neurocognitive disorders (HAND). In this review we briefly summarize the molecular mechanisms involved in the pathological production of the toxic sphingolipid, ceramide and address questions of how cytokine and cellular stress pathways that are perturbed in HAND converge to deregulate ceramide-associated signaling. PMID- 18508576 TI - Resistance mechanisms to cancer chemotherapy. AB - Resistance to chemotherapy ('drug resistance') is a fundamental problem that limits the effectiveness of many chemotherapies currently used to treat cancer. Drug resistance can occur due to a variety of mechanisms, such as increased drug inactivation, drug efflux from cancer cells, enhanced repair of chemotherapy induced damage, activation of pro-survival pathways and inactivation of cell death pathways. In this article, we review some of the major mechanisms of drug resistance and discuss how new molecularly-targeted therapies are being increasingly used to overcome these resistance mechanisms. PMID- 18508577 TI - Genetic background in apolipoprotein A-I and cystathionine beta-synthase deficiency. AB - Double heterozygous mice lacking one allele of Cbs and Apoa1 develop hyperhomocysteinemia and hypoalphalipoproteinemia together with moderate hypertension. To study the influence of the genetic background into this specific phenotype, four groups of male mice were established: control and double heterozygous groups in C57BL/6J and in C57BL/6J x 129 backgrounds, respectively. Nitric oxide levels, systolic blood pressure, plasma lipid parameters, arylesterase activity and aorta histology were analyzed as well as oligonucleotide array hybridization of liver RNA. Results demonstrated that double heterozygous mice in C57BL/6J substrate had a milder phenotype showing lower increase in blood pressure compared to double heterozygous group in hybrid background. The severity of the phenotype in the latter group was associated with lower nitric oxide and arylesterase activity levels, and hyperplasia of the vascular media layer. Hepatic profiling of both genetic substrates showed profound differences in expression of contractile proteins that could explain these pathological findings. In summary, the phenotypic presentation of hypertension is associated with multiple processes from vascular bedside to liver as evidenced by nitric oxide production or paraoxonase levels. PMID- 18508578 TI - Bone marrow-derived progenitor cells and renal fibrosis. AB - Bone marrow-derived cells modulate solid organ diseases. For example, the different immune cell populations mediate tissue inflammation and damage, whereas progenitor cell populations are thought to enhance tissue regeneration. However, in the process of tissue fibrosis the contribution of either cell type is less clear. Here we discuss current concepts on how different bone marrow-derived progenitor cell populations may contribute to renal fibrosis. PMID- 18508579 TI - Novel actions of tissue-type plasminogen activator in chronic kidney disease. AB - Tissue-type plasminogen activator (tPA) is traditionally viewed as a simple serine protease whose main function is to convert plasminogen into biologically active plasmin. As a protease, tPA plays a crucial role in regulating blood fibrinolysis, in maintaining the homeostasis of extracellular matrix and in modulating the post-translational activation of growth factors. However, emerging evidence indicates that tPA also functions as a cytokine that transmits its signal across the cell membrane, initiates a diverse array of intracellular signaling, and dictates gene expression in the nuclei. tPA binds to the cell membrane LDL receptor-related protein 1 (LRP-1), triggers its tyrosine phosphorylation. As a cytokine, tPA plays a pivotal role in the pathogenesis of renal interstitial fibrosis through diverse mechanisms. It facilitates tubular epithelial to mesenchymal transition, potentiates myofibroblast activation, and protects renal interstitial fibroblasts/myofibroblasts from apoptosis. Together, growing evidence has implicated tPA as a fibrogenic cytokine that promotes the progression of kidney diseases. These new findings have radically changed our conception of tPA in renal fibrogenesis and represent a paradigm shift towards uncovering its cytokine function. PMID- 18508580 TI - The implication of proinflammatory cytokines in type 2 diabetes. AB - The incidence of type 2 diabetes (T2D) is rapidly expanding. Some of the more obvious pathologies associated with it include: defective glucose metabolism, obesity, cardiovascular disease and an inability to mount an effective immune response to infection by certain pathogenic organisms, leading to sepsis and death. A common tie linking these seemingly disparate complications is chronic inflammation. Today we know that inflammation is regulated locally and systemically by numerous biochemical signals. One of the most important of these signals is a class of molecules called cytokines. Cytokines can be generally classified as proinflammatory or anti-inflammatory and allow an organism to respond rapidly to an immune challenge by coordinating an appropriate immune response. In T2D, the balance between proinflammatory and anti-inflammatory cytokines is shifted toward proinflammation, potentially causing or exacerbating the health complications found in T2D. Over-nutrition has been shown to trigger the innate immune system but activation of the innate immune system, itself, induces hyperglycemia and insulin resistance. In all likelihood, diabetes and chronic inflammation are inseparable and act as a reciprocal feed-forward loop. PMID- 18508581 TI - Glycosylation is crucial for stability of tumour and cancer stem cell antigen EpCAM. AB - Epithelial cell adhesion molecule EpCAM is strongly over-expressed in a variety of carcinomas where it is involved in signalling events resulting in increased expression of target genes such as c-Myc, cyclins and others, eventually conferring cells an oncogenic phenotype. However, EpCAM is also expressed in a series of healthy epithelia, albeit generally to a far lesser extend. We have uncovered differential glycosylation of EpCAM as a means to discriminate normal from malignant tissues. EpCAM was hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. All three N-glycosylation consensus sequences within EpCAM's extracellular domain were used in human and murine cells. We show that glycosylation at asparagine198 is crucial for protein stability. Mutants of EpCAM that substitute asparagine198 for alanine showed a decreased overall expression and half-life of the molecule at the plasma membrane. This is of considerable importance with respect to EpCAM variants expressed in normal tissue, where it might reveal to be less stable and thus may have repercussions on functionality. PMID- 18508582 TI - The Arabidopsis checkpoint protein Bub3.1 is essential for gametophyte development. AB - Except for Mad2 homologues of maize and wheat, no spindle checkpoint proteins which regulate the onset of anaphase during nuclear divisions have been reported for plants so far. We found three homologues of the spindle checkpoint protein Bub3 in Arabidopsis thaliana. Bub3.1 and Bub3.2 are 88 percent identical at the amino acid level, while Bub3.3 is more distantly related. Bub3.1 and Bub3.2 mRNA appeared preferentially in mitotically active tissues. Bub3.1 but not Bub3.2 showed increased expression during mitosis. Nuclear but no centromeric localization was shown for EYFP-Bub3.1 and Bub3.1-ECFP fusion constructs. Three T DNA insertion mutants each were identified for Bub3.1 and Bub3.2 encoding genes. One bub3.1 mutant survived only in heterozygous state and displayed defects in development of male and female gametophytes, while two other homozygous bub3.1 mutants revealed transcripts and are therefore no null mutants. In contrast, all three homozygous bub3.2 mutants appeared to be viable and fertile without generating full-length transcripts. PMID- 18508583 TI - Origins and evolution of modern biochemistry: insights from genomes and molecular structure. AB - The survey of components in living systems at different levels of organization enables an evolutionary exploration of patterns and processes in macromolecules, networks, and genomic repertoires. Here we discuss how phylogenetic strategies that generate intrinsically rooted phylogenies impact the evolutionary study of RNA and protein components of the macromolecular machinery that is responsible for biological function. We used these methods to generate timelines of discovery of components in systems, such as substructures in RNA molecules, architectures in proteomes, domains in multi-domain proteins, enzymes in metabolic networks, and protein architectures in proteomes. These timelines unfolded remarkable patterns of origin and evolution of molecules, repertoires and networks, showing episodes of both functional specialization (e.g., rise of domains with specialized functions) and molecular simplification (e.g., reductive tendencies in molecules and proteomes). These observations have important evolutionary implications for origins of translation, the genetic code, modules in the protein world, and diversification of life, and suggest early evolution of modern biochemistry was driven by recruitment of both RNA and protein catalysts in an ancient community of complex organisms. PMID- 18508584 TI - The many roles of the regulatory protein ICP27 during herpes simplex virus infection. AB - Herpes simplex virus type 1 (HSV-1) protein ICP27 is a multifunctional regulator of gene expression that assumes different roles during the course of infection. Early in infection, ICP27 mediates the inhibition of cellular splicing, whereas, later it helps to recruit cellular RNA polymerase II (RNAP II) to viral replication sites and to facilitate viral RNA export. ICP27 has also been shown to stimulate translation of viral transcripts. ICP27 performs its activities by interacting with RNA and with an assortment of proteins. ICP27 binds viral RNAs in its role as an export adaptor. An ever increasing number of cellular proteins have been shown to interact with ICP27, including splicing factors, export proteins and RNAP II. A number of protein motifs within ICP27 have been predicted based upon sequence comparisons; however, detailed structural information is not yet available. Although much has been learned about the mechanisms by which ICP27 performs its roles, relatively little is known about how its activities are regulated. The roles and activities of ICP27 are the subject of this review. PMID- 18508585 TI - RAS and PKA pathways in cancer: new insight from transcriptional analysis. AB - Through its ability to regulate the activity of a large number of transcription factors, the Ras pathway is able to control several transcriptional programs leading to proliferation, differentiation, metabolism, cytoskeletal reorganization and immune response. Cyclic AMP (cAMP) is a ubiquitous intracellular second messenger whose major intracellular target in eukaryotes is protein kinase A (PKA). Wide evidence for cross talk between the Ras and cAMP-PKA pathways is available. After reviewing some features of Ras and PKA signalling that are relevant for cancer biology, we re-analyze available genome-wide expression data for genes encoding proteins of the downstream branch of the PKA pathway in human tumor cell lines as a function of the mutational state of the Ras pathway. The observed Ras-dependent pattern of regulation of the analyzed genes may contribute to explain how the cAMP-PKA axis is involved in oncogenic processes induced by Ras. PMID- 18508586 TI - The redundant-signals paradigm and preattentive visual processing. AB - Physiological and cognitive models of vision agree that the pre-attentive processing of visual stimuli is organized in a parallel and segregated fashion. However, several incompatible models have been proposed for the subsequent processing stages. They differ in their assumptions about architecture (serial, parallel, or coactive/integrative), stopping-rules (self-terminating, or exhaustive), spatial specificity of saliency signal coding (signal pooling across locations, or spatially distinct processing), and dependency of target detection on the prior allocation of attention (pre-attentive, or post-selective). We review how studies employing the redundant-signals paradigm in visual pop-out search contribute to discerning between the different assumptions. We find strong support for the notion of a saliency map, into which feature contrast signals are pooled, and especially the dimension weighting account (1) receives further support: Instead of a priming mechanism that could increase weights for several dimensions independently, evidence favors a weighting mechanism that effectively limits the total weight available for allocation to the various dimensions through competitive interactions, whereby increasing the weight for one dimension goes along with decreasing the weights for other dimensions. PMID- 18508587 TI - Mannose polymer induces vasodilation through a luminal mannose receptor in rat mesenteric arteries. AB - Several of the luminal endothelial glycocalyx functions are exerted via interactions with glycosidic components and sugar binding proteins with lectinic activity. One important example is the mannose receptor (MR). The MR has been detected in cell types that mediate the phagocytosis and pinocytosis of particles and solutes containing mannose. Using isolated constant pressurized rat mesenteric arteries (RMA), we evaluated the effects of a mannose polymer in the vascular tone. RMA were pre-contracted with 10 micromol/L phenylephrine and carbohydrates were perfused at 20 microliters/min. Perfusion of free D-mannose (1 nmol/L to 100 micromol/L) induced a concentration-dependent vasodilation of pre contracted RMA. Perfusion of mannose polymer (1 nmol/L to 100 micromol/L) induced a larger effect in a concentration-dependent vasodilation. Mannose polymer's maximum effect reached a 96 percent of basal diameter; this significant vasodilation was not nitric oxide (NO) or cyclooxygenase (COX) dependent effect. We corroborated the binding of the mannose polymer to the endothelial lumen, by perfusion of a fluorescently labeled mannose polymer; and also, we detected a significant level of MR mRNA in whole mesenteric arteries. With all these, we proposed a novel effect of a MR in the regulation of vascular tone. PMID- 18508588 TI - The modulatory capacity of interleukin-21 in the pathogenesis of autoimmune disease. AB - In this review, we discuss recent progress from studies on the biology of IL-21 and the role of this cytokine in the pathogenesis of autoimmunity. Recent studies have demonstrated that IL-21 plays an important and non-redundant role in a number of autoimmune animal models indicating that IL-21 could be a common modulator of the adaptive immune response towards self-tissue constituents in diseases such as systemic lupus erythematosus, models of rheumatoid arthritis, multiple sclerosis and type-1 diabetes. Also, the studies on the production of IL 21 in human autoimmune diseases and its behaviour on human cells in vitro are revealing the potential of IL-21 to exacerbate cellular processes that determine the course of autoimmune diseases. PMID- 18508589 TI - Does deep brain stimulation induce apathy in Parkinson's disease? AB - Chronic deep brain stimulation (DBS) is an important therapeutic advancement for the treatment of motor symptoms in Parkinson's disease (PD). However, its effects on non-motor symptoms are not well understood. Several studies have reported motivational disturbances and apathy after DBS surgery. Recent studies are beginning to more carefully examine the relationship between DBS and apathy. This review summarizes and evaluates the current state of the literature on apathy after DBS surgery, discusses methodological limitations in the literature, and makes suggestions for future research. PMID- 18508591 TI - Cellular and molecular mechanisms of resistance to oral Candida albicans infections. AB - Oral candidiasis is a significant health problem in terms of both morbidity and economic outlay. Infections are predominantly caused by the commensal C. albicans, and affect immunocompromised individuals, including HIV-positive and AIDS patients, organ transplant recipients and chemotherapy patients. The molecular and cellular immune mechanisms involved in protection from and responses to oral candidiasis are overlapping, but distinct from those associated with other manifestations of the disease, including systemic, vaginal and gastric candidiasis. In oral candidiasis, clinical observations and experimental mouse models suggest a critical role for cell-mediated immunity. In mice, CD4+ T-cells and the p40 subunit of interleukins 12 and 23 are strict prerequisites for resistance; however abrogation of IFN-gamma does not confer susceptibility. Here, we discuss this apparent inconsistency, and review the experimental evidence that clarifies which immune pathways are specifically involved in resistance and responses to candidiasis of the oral cavity. We also highlight deficiencies in the literature, particularly concerning the putative roles of some relatively new elements in immunobiology: interleukin-23, interleukin-17 and T helper (Th)17 cells. PMID- 18508592 TI - The Mnks: MAP kinase-interacting kinases (MAP kinase signal-integrating kinases). AB - The human MAP kinase-interacting kinases (or MAP kinase signal-integrating kinases), Mnks, comprise a group of four proteins derived from two genes (Gene symbols: MKNK1 and MKNK2) by alternative splicing. Mnk1a/b differ at their C termini, as do Mnk2a/2b: in each case, the a-form possesses a longer C-terminal region than the b-form, which lacks the MAP kinase-binding region. The N-termini of all forms contain a polybasic region which binds importin a and the translation factor scaffold protein eukaryotic initiation factor (eIF) 4G. The catalytic domains of Mnk1a/b and Mnk2a/b share three unusual features: two short inserts and a DFD feature where other kinases have DFG. Mnk isoforms differ markedly in their activity and regulation, and in subcellular localization. The best-characterised Mnk substrate is eIF4E. The cellular role of eIF4E phosphorylation remains unclear: it may promote export of certain mRNAs from the nucleus. Other Mnk substrates bind to AU-rich elements that modulate the stability/translation of specific mRNAs. Mnks may also control production of inflammatory mediators and signaling from tyrosine kinase receptors, as well as cell proliferation or survival. PMID- 18508593 TI - TNF: a moonlighting protein at the interface between cancer and infection. AB - The remarkable ability of TNF, especially in combination with Interferon-gamma or melphalan, to inhibit the growth of malignant tumor cells is so far unmatched. Unfortunately, its high systemic toxicity and hepatotoxicity prevent its systemic use in cancer patients. An elegant manner to circumvent this problem is the isolated limb and liver perfusion for the treatment of melanoma, soft tissue sarcoma and liver tumors, respectively, although the latter method can lead to a reversible hepatotoxicity. In order to allow also the treatment of other cancers with TNF, new strategies have to be developed that aim at sensitizing tumor cells to TNF and at reducing its systemic and liver toxicity, without losing its antitumor efficiency. Moreover, the lectin-like domain of TNF, which is spatially distinct from the receptor binding sites, could be useful in reducing cancer treatment-related pulmonary edema formation. This review will discuss some recent developments in these areas, which can lead to a renewed interest in TNF for the systemic treatment of cancer. PMID- 18508590 TI - Role of oxidative stress and nitric oxide in atherothrombosis. AB - During the last decade basic and clinical research has highlighted the central role of reactive oxygen species (ROS) in cardiovascular disease. Enhanced production or attenuated degradation of ROS leads to oxidative stress, a process that affects endothelial and vascular function, and contributes to vascular disease. Nitric oxide (NO), a product of the normal endothelium, is a principal determinant of normal endothelial and vascular function. In states of inflammation, NO production by the vasculature increases considerably and, in conjunction with other ROS, contributes to oxidative stress. This review examines the role of oxidative stress and NO in mechanisms of endothelial and vascular dysfunction with an emphasis on atherothrombosis. PMID- 18508594 TI - Apolipoprotein E may be a critical factor in hormone therapy neuroprotection. AB - In this review we examine the evidence for ovarian hormone neuroprotection in chronic neurological diseases, including stroke. We propose that neuroprotection may involve the ability of estrogens to modulate apolipoprotein E (apoE) and its receptor, the low density lipoprotein receptor related protein (LRP). Results from numerous studies have demonstrated that (1) nerve regeneration is severely delayed in apoE-gene knockout (KO) mice as compared to wild-type (WT) littermates; (2) 17beta estradiol replacement in ovariectomized mice resulted in a significant increase in levels of apoE and LRP, in the olfactory bulb (OB) and other brain areas; (3) estradiol treatment increased both apoE and neurite outgrowth in cortical and olfactory neuronal cultures; and (4) estradiol treatment had no effect on neurite outgrowth in cultures deprived of apoE or in the presence of apoE4. In essence these studies suggest that apoE is a critical intermediary for the beneficial effects of 17beta estradiol on nerve repair, which can lead to functional reorganization (plasticity). Future studies of HT should evaluate the effects of apoE genotype and production estradiol on neuroprotection. PMID- 18508595 TI - Cystatins: biochemical and structural properties, and medical relevance. AB - The cystatin superfamily comprises a large group of the cystatin domain containing proteins, present in a wide variety of organisms, including humans. Cystatin inhibitory activity is vital for the delicate regulation of normal physiological processes by limiting the potentially highly destructive activity of their target proteases such as the papain (C1) family, including cysteine cathepsins. Some of the cystatins also inhibit the legumain (C13) family of enzymes. Failures in biological mechanisms controlling protease activities result in many diseases such as neurodegeneration, cardiovascular diseases, osteoporosis, arthritis, and cancer. Cystatins have been classified into three types: the stefins, the cystatins and the kininogens, although other cystatin related proteins, such as CRES proteins, are emerging. The stefins are mainly intracellular proteins, whereas the cystatins and the kininogens are extracellular. The cystatins are tight binding and reversible inhibitors. The basic mechanism of interaction between cystatins and their target proteases has been established, based mainly on the crystal structures of various cathepsins, stefins and cystatins and their enzyme-inhibitor complexes. Cystatins, as rather non-selective inhibitors, discriminate only slightly between endo- and exopeptidases. They are also prone to form amyloids. The levels of some stefins and cystatins in tissue and body fluids can serve as relatively reliable markers for a variety of diseases. In this review we summarize present knowledge about cystatins and their role in some diseases. PMID- 18508596 TI - Involvement of endothelial Man and Gal-binding lectins in sensing the flow in coronary arteries. AB - The coronary endothelial luminal membrane (CELM) glycocalyx has diverse molecules involved in blood flow signal transduction. Evidence suggests that some of these structures may be lectinic. To test this, we synthesized two monosaccharide polymers (Mon-Pols) made of Mannose (Man-Pol) or Galactose (Gal-Pol) covalently coupled to Dextran (70 kDa) and used them as lectin affinity probes. In situ intracoronary infusion of both polymers resulted in CELM-binding but only Man-Pol caused a reduction in flow-induced positive inotropism and dromotropism. To demonstrate that our lectinic probes could bind to CELM lectins, a representative CELM protein fraction was isolated via intracoronary infusion of a cationic silica colloid and either Mannose- or Galactose-binding lectins were purified from the CELM protein fraction using the corresponding Mon-Pol affinity chromatography resin. Resin-bound CELM proteins were eluted with the corresponding monosaccharide. 2D-SDS-PAGE (pH 4-7) revealed 9 Mannose- and approximately 100 Galactose-selective CELM lectins. In summary, the CELM glycocalyx contains Mannose- and Galactose-binding lectins that may be involved in translating coronary flow into a cardiac parenchymal response. PMID- 18508597 TI - Rapid detection of botulinum neurotoxins A, B, E, and F by optical immunoassay. AB - Botulism is caused by the neurotoxins (BoNTs) produced from Clostridium botulinum. These neurotxins often lead to fatal neuroparalytic disease which is regarded a major threat to the public health. For this reason, rapid and reliable diagnosis of BoNTs in field settings and peripheral care centers is highly valuable. Here, we describe a multiplexed and sensitive optical immunoassay (OIA) for the rapid detection of four medically important BoNTs (A, B, E, and F). The assay is based on detection of physical changes in the thickness of molecular thin film resulting from specific immunobinding events on an optical silicon chip. The immunocomplex causes destructive interference of a particular wavelength of reflected white light from gold to purple-blue on an optical surface depending on the concentration of the analyte. This test allows simultaneous characterization of toxin type and semi-quantitative assessment of toxin level. The assay was four times more sensitive than ELISA when performed with the same reagents. The limit of detection (LOD) for the BoNTs was, respectively, 2.5-5 ng/mL, 5-10 ng/mL, and 10-20 ng/mL in experimentally spiked buffer, water and food matrices. The less logistic load and visual read-out of this method promises potential applicability of this assay in the field as well as in a clinical settings. PMID- 18508598 TI - Structure and ligand interactions of the urokinase receptor (uPAR). AB - The urokinase-type plasminogen activator receptor (uPAR or CD87) is a glycolipid anchored membrane glycoprotein, which is responsible for focalizing plasminogen activation to the cell surface through its high-affinity binding to the serine protease uPA. This tight interaction (KD less than 1 nM) is accomplished by an unusually large and hydrophobic binding cavity in uPAR that is created by a unique interdomain assembly involving all three homologous domains of the receptor. These domains belong to the Ly-6/uPAR (LU) protein domain family, which is defined by a consensus sequence predominantly based on disulfide connectivities, and they adopt a characteristic three-finger fold. Interestingly, the gene for uPAR is localized in a cluster of 6 homologous genes encoding proteins with multiple LU-domains. The structural biology of uPAR will be reviewed with special emphasis on its multidomain composition and the interaction with its natural protein ligands, i.e. the serine protease uPA and the matrix protein vitronectin. PMID- 18508599 TI - Urokinase and its receptors in chronic kidney disease. AB - This review focuses on the role of the serine protease urokinase-type plasminogen activator and its high affinity receptor uPAR/CD87 in chronic kidney disease (CKD) progression. An emerging theme is their organ- and site-specific effects. In addition to tubules, uPA is produced by macrophages and fibroblasts in CKD. By activating hepatocyte growth factor and degrading fibrinogen uPA may have anti fibrotic effects. However renal fibrosis was similar between uPA wild-type and knockout mice in experimental CKD. The uPAR is expressed by renal parenchymal cells and inflammatory cells in a variety of kidney diseases. Such expression appears anti-fibrotic based on studies in uPAR-deficient mice. In CKD uPAR expression is associated with higher uPA activity but its most important effect appears to be due to effects on cell recruitment and migration that involve interactions with a variety of co-receptors and chemoattractant effects of soluble uPAR. Vitronectin and high molecular weight kininogen are alternate uPAR ligands, and receptors in addition to uPAR may also bind directly to uPA and activate cell signaling pathways. PMID- 18508601 TI - MK2 and MK3--a pair of isoenzymes? AB - The MAPK-activated protein kinases MK2 and MK3 form a pair of structurally and functionally closely related enzymes present in mammals and birds. Both protein kinases can bind to p38alpha MAPK and are activated by p38alpha via multiple proline-directed phosphorylations in a stress-dependent manner. Although the expression level and activity of MK2 is always significantly higher than that of MK3, the substrate spectrum of both enzymes is indistinguishable and covers proteins involved in cytokines production, endocytosis, reorganization of the cytoskeleton, cell migration, cell cycle control, chromatin remodeling and transcriptional regulation. Functional differences between MK2 and MK3 could result from the more prominent proline-rich SH3-targeting region in MK2, but are not reported so far. Since MK2 and MK3 are the main downstream targets of p38alpha responsible for posttranscriptional stimulation of cytokine biosynthesis, both enzymes are promising targets for the development of small molecule inhibitors which can be used in anti-inflammatory therapy. MK2-knockout mice show decreased LPS-induced cytokine biosynthesis and increased protection against collagen-induced arthritis. Recently generated MK2/3 double knockout mice show further reduction of LPS-induced cytokine production. PMID- 18508602 TI - ATP-dependent chromatin remodeling enzymes and their various roles in cell cycle control. AB - The modification of chromatin structure by various mechanisms has emerged as a key regulatory component of nuclear programs. Cell cycle progression and exit are affected by the integrity of chromatin architecture as well as by regulatory cues that chromatin structure imposes on the expression of cell cycle genes. ATP dependent chromatin remodeling factors use the energy derived from ATP-hydrolysis to modulate histone-DNA contacts. These molecular machines play important roles in all aspects of chromosome biology and are thus intimately linked to cell cycle control. Regulation of complex activity by various signaling pathways has been a rising theme in recent years. Moreover, some chromatin remodeling factors have been characterized as potent tumor suppressor proteins. Thus, to understand the functions and activities of ATP-utilizing chromatin remodeling factors is an important goal towards their use as potential targets in cancer therapy. PMID- 18508603 TI - Posttranscriptional control of HBV gene expression. AB - Acute and chronic hepatitis B virus (HBV) infection is the cause of about 1 million deaths each year worldwide. Although a vaccine for HBV is available, new HBV infections are appearing at alarming rates and maternal-fetal transmission is a major cause of viral spread. Thus, new and effective antiviral strategies are necessary for the 300 million chronically HBV-infected individuals to reduce their morbidity and mortality. Precise processing of viral RNAs is essential for the hepatitis viral life cycle, and, to our knowledge, the processing, nuclear export, and stabilization/degradation of viral RNAs are exclusively mediated by host factors. Thus, identification of host factors required for viral RNA metabolism and subsequent molecular analysis of the interactions between viral RNAs and corresponding host factors might represent novel avenues for the development of new antiviral strategies. In this review, we summarize the current knowledge about the posttranscriptional control of HBV RNA metabolism. PMID- 18508604 TI - Novel methods for detecting amyloidogenic proteins in transthyretin related amyloidosis. AB - Transthyretin (TTR)-related familial amyloidotic polyneuropathy (FAP) is an autosomal dominant form of fatal hereditary amyloidosis. Until 25 years ago, tools for diagnosis of FAP were restricted to clinical manifestations and pathologic methods, and a small number of patients in the restricted endemic areas could be diagnosed with this disease. However, owing to progress in biochemical and molecular genetic analyses, this disease is now believed to occur worldwide. As of today, reports of about 100 different points of single or double mutations, or a deletion in the TTR gene have been published, and several different phenotypes of FAP have been documented, even for the same mutation in the TTR gene. Since liver transplantation has been established to halt the progression of FAP, rapid and reliable diagnostic system for FAP is needed. We present here a new diagnostic procedure for the disease using current methods of molecular genetics and protein chemistry. PMID- 18508605 TI - Control of granule exocytosis in neutrophils. AB - Neutrophils are granulocytes derived from bone marrow that circulate through the blood and become recruited to tissues during infection or inflammation. They are the most abundant white blood cell and comprise the first line of defence in the innate immune system. However, they are also capable of causing tissue damage in a wide range of diseases. Release of chemotactic signals from inflamed or infected tissues trigger neutrophil migration from the bloodstream to inflammatory foci, where they contribute to inflammation by undergoing receptor mediated respiratory burst and degranulation. Degranulation from neutrophils has been implicated as a major causative factor in numerous inflammatory diseases. However, the mechanisms that control neutrophil degranulation are not well understood. Recent observations indicate that receptor-mediated granule release from neutrophils depends on activation of distal signaling pathways that include the src family of tyrosine kinases, beta-arrestins, the tyrosine phosphatase MEG2, the kinase MARCK, Rabs and SNAREs, and the Rho GTPase, Rac2. Some of these pathways are specifically required for membrane fusion between the granule and plasma membrane, leading to exocytosis. This review focuses on the understanding of distal molecular mechanisms controlling exocytosis from neutrophils. PMID- 18508600 TI - Structural answers and persistent questions about how nicotinic receptors work. AB - The electron diffraction structure of nicotinic acetylcholine receptor (nAChR) from Torpedo marmorata and the X-ray crystallographic structure of acetylcholine binding protein (AChBP) are providing new answers to persistent questions about how nAChRs function as biophysical machines and as participants in cellular and systems physiology. New high-resolution information about nAChR structures might come from advances in crystallography and NMR, from extracellular domain nAChRs as high fidelity models, and from prokaryotic nicotinoid proteins. At the level of biophysics, structures of different nAChRs with different pharmacological profiles and kinetics will help describe how agonists and antagonists bind to orthosteric binding sites, how allosteric modulators affect function by binding outside these sites, how nAChRs control ion flow, and how large cytoplasmic domains affect function. At the level of cellular and systems physiology, structures of nAChRs will help characterize interactions with other cellular components, including lipids and trafficking and signaling proteins, and contribute to understanding the roles of nAChRs in addiction, neurodegeneration, and mental illness. Understanding nAChRs at an atomic level will be important for designing interventions for these pathologies. PMID- 18508606 TI - Host CD147 blockade by small interfering RNAs suppresses growth of human colon cancer xenografts. AB - Tumor cells can stimulate matrix metalloproteinase (MMP) production by stromal cells through cell-cell interactions mediated by cell adhesion molecules such as extracellular matrix metalloproteinase inducer (human CD147/EMMPRIN, mouse CD147/Basigin). This study sought to characterize whether specific tumor-stromal cell interactions mediated by CD147 promote colon cancer growth by utilizing small interfering (si)RNAs directed against human CD147/EMMPRIN or mouse CD147/Basigin in co-cultures of cancer cells with macrophages and fibroblasts and established human SW620 colon cancer xenograft models in immune deficient mice. We show that blockade of host (mouse) CD147/Basigin expression, but not cancer cell-derived CD147/EMMPRIN, suppresses tumor growth in human colon cancer xenografts. Experiments in vitro indicated that colon cancer cell-stromal cell interactions mediated by CD147 lead to increased MMP-2 expression in fibroblasts but not macrophages. Furthermore, expression of host VEGF-A in both fibroblasts and macrophages is independent of CD147 in vitro and in vivo. Interestingly, inhibition of cancer cell-derived EMMPRIN leads to increased MMP-9 levels in vivo. Our findings provide new insights into CD147-mediated tumor-host interactions mediating colon cancer growth. PMID- 18508607 TI - Modulation of tumor associated macrophages in solid tumors. AB - It is becoming increasingly clear that interactions between cancer cells, stroma cells and the extracellular matrix (ECM) are pivotal to the processes of neovascularization and tumorigenesis. As tumor stromal components known as tumor associated macrophages (TAMs), mononuclear phagocytes can play a key role in tumor type specific neoangiogenesis by promoting remodeling of the ECM through the production of matrix metalloproteases (MMPs), secreting pro-angiogenic growth factors and stabilizing the tumor vasculature. The growth factor colony stimulating factor-1 is produced by a wide variety of cancer cells and tumor stroma cells and influences the migration, survival and phenotype of TAMs. Thus, understanding the relationships of the cancer cell with the host environment is key for specifically exploiting tumor growth promoting tumor host interactions for new therapeutic strategies. This review outlines the strategies for targeting CSF-1 in malignancies to influence TAMs in tumor development. PMID- 18508608 TI - How is AMPK activity regulated in skeletal muscles during exercise? AB - AMPK is a metabolic "master" controller activated in skeletal muscle by exercise in a time and intensity dependent manner, and has been implicated in regulating metabolic pathways in muscle during physical exercise. AMPK signaling in skeletal muscle is regulated by several systemic and intracellular factors and the regulation of skeletal muscle AMPK in response to exercise is the focus of this review. Specifically, the role of LKB1 and phosphatase PP2C in nucleotide dependent activation of AMPK, and ionized calcium in CaMKK-dependent activation of AMPK in working muscle is discussed. We also discuss the influence of reactive oxygen species produced within the muscle as well as muscle glycogen and TAK1 in regulating AMPK during exercise. Currently, during intensive contraction, activation of alpha2-AMPK seems mainly to rely on AMP accumulating from ATP hydrolysis whereas calcium signaling may have some importance during more gentle contraction conditions. Factors that regulate alpha1-AMPK during exercise are less clear but it appears, at least to some extent, to rely on an adenine nucleotide-dependent mechanism. PMID- 18508609 TI - The genomics of LUCA. AB - To understand the nature and evolution of LUCA, or Last Universal Common Ancestor, the minimum genome of LUCA has been identified based on the genes common to the eight primitive Euryarchaea and Crenarchaea species Methanopyrus kandleri, Methanothermobacter thermautotrophicum, Methanococcus jannaschii, Pyrococcus abyssi, Pyrococcus furiosus, Pyrococcus horikoshii, Aeropyrum pernix and Pyrobaculum aerophilum, together with the methanogenesis genes of the primitive methanogens. The 424 protein encoding genes in the minimum LUCA genome exceed significantly the 150-340 genes estimated to be present in a minimal proteome compatible with life. Thus LUCA was not a minimal organism but the first modern organism equipped with a DNA genome and the universal genetic code. The hyperthermophilic, Methanopyrus-proximal LUCA is consistent with a Hot Cross Origin of life which proposes that early heterotrophic life forms in the cooler temperature zones invented methanogenesis and a DNA genome upon their adaptation to the hydrothermal vents, where life flourished massively on lithoautotrophy supported by carbon dioxide and hydrogen, thereby leading to the rise of LUCA. PMID- 18508610 TI - Calculation of the free energy barriers in the oligomerisation of Abeta peptide fragments. AB - Protein misfolding and aggregation are associated with a range of severe human neurodegenerative conditions. We use all-atom simulations to describe the process of assembly of the Abeta(16-22) and Abeta(25-35) fragments of Abeta, a peptide associated with Alzheimer's disease. Our results indicate that the pathways of aggregation of these two peptides depend predominantly on the relative strength of hydrophobic interactions and hydrogen bonding. In the Abeta(25-35) peptide, which is weakly hydrophobic, the tendency to form hydrogen bonds drives the crossing of a single major free energy barrier for the formation of a cross-beta structure. By contrast, in the more hydrophobic Abeta(16-22) peptide, the process of ordered assembly is preceded by an initial collapse into disordered oligomers. These results provide support for a recently proposed two-step mechanism of amyloid formation. We have also found that the barriers for reordering are lower for large oligomers than for small oligomers, a result that provides an explanation of the recent experimental observation that the efficiency of the seeding reaction depends on the size of the seeds themselves. PMID- 18508611 TI - Ca2+ signalling in the control of motility and guidance in mammalian sperm. AB - Ca2+ signalling in the sperm plays a key role in the regulation of events preceding fertilisation. Control of motility, including hyperactivation and chemotaxis, is particularly dependent upon [Ca2+]i signalling in the principal piece of the flagellum and the midpiece. Here we briefly review the processes that contribute to regulation of [Ca2+]i in mammalian sperm and then examine two areas: (i) the regulation of hyperactivation by [Ca2+]i and the pivotal roles played by CatSpers (sperm-specific, Ca2+-permeable membrane channels) and intracellular Ca2+ stores in this process and (ii) the elevation of [Ca2+]i and consequent modulation of motility caused by progesterone including the ability of progesterone at micromolar concentrations to cause sperm hyperactivation and/or accumulation and the recent discovery that progesterone, at picomolar concentrations, acts as a chemoattractant for mammalian sperm.. PMID- 18508612 TI - Reversal of effects of acidosis on contraction of rat heart myocytes by CGP 48506. AB - In experiments reported here, we tested the ability of CGP-48506 to reverse the depressed cardiac contractility associated with hypercapnic acidosis in isolated rat cardiac myocytes. CGP-48506 is a cardiotonic agent that directly and specifically promotes the actin-cross-bridge reaction. Myocytes superfused at pH 6.8 demonstrated a significantly reduced extent of cell shortening, but an increase in the peak amplitude of the Ca2+ transient. Moreover, cells in acidosis showed small, but significant, decreases in time to peak shortening to 50 percent relaxation. Superfusion of the cells with 3, 7, and 10 micro-molar CGP-48506 restored the inhibited contractility as a function of concentration with no significant effects on the Ca2+-transient. Moreover, 10 micro-molar CGP-48506 completely reversed the depressed myocyte contraction associated with an increase in time to peak shortening and time to 50 percent and 75 percent relaxation. Our results indicate that the depression of contractility associated with acidosis is due to a reduced myofilament response to Ca2+, which can be overcome by agents working downstream from troponin C through a direct effect on the actin-myosin interaction. PMID- 18508613 TI - Papillomavirus 3' UTR regulatory elements. AB - Papillomaviruses infect epithelial cells causing mainly benign lesions or warts. In some rare instances, these may progress to malignancy. For example, human papillomavirus type 16 (HPV16) is the causative agent of 60 percent of cases of cervical cancer. The replication cycle of papillomaviruses is intimately linked to epithelial differentiation. In particular, late gene expression is completed exclusively in the upper epithelial layers. Regulation of late gene expression is largely by post-transcriptional means. RNAs encoding the late proteins, the virus capsid proteins L1 and L2, can be detected in the lower layers of infected epithelia but protein is detected only in the upper layers. It is clear that cellular factors mediate this gene regulation. RNA regulatory elements that bind RNA processing factors that mediate post-transcriptional control have been identified in the 3' untranslated regions of a number of papillomaviruses. These elements, the proteins they bind and the mechanisms by which they are proposed to act are discussed. Further understanding of such host-virus molecular interactions may lead to development of novel strategies for abrogating virus infection. PMID- 18508615 TI - Self-assembly of amyloid-forming peptides by molecular dynamics simulations. AB - Protein aggregation is associated with many neurodegenerative diseases. Understanding the aggregation mechanisms is a fundamental step in order to design rational drugs interfering with the toxic intermediates. This self-assembly process is however difficult to observe experimentally, which gives simulations an important role in resolving this problem. This study shows how we can proceed to gain knowledge about the first steps of aggregation. We first start by characterizing the free energy surface of the Abeta (16-22) dimer, a well-studied system numerically, using molecular dynamics simulations with OPEP coarse-grained force field. We then turn to the study of the NHVTLSQ peptide in 4-mers and 16 mers, extracting information on the onset of aggregation. In particular, the simulations indicate that the peptides are mostly random coil at room temperature, but can visit diverse amyloid-competent topologies, albeit with a low probability. The fact that the 16-mers constantly move from one structure to another is consistent with the long lag phase measured experimentally, but the rare critical steps leading to the rapid formation of amyloid fibrils still remain to be determined. PMID- 18508616 TI - Modulating HIV-1 RNA processing and utilization. AB - Expression of the integrated HIV-1 provirus is achieved by overcoming multiple barriers to the processing, transport and utilization of the viral RNA. Some of the strategies involve viral encoded proteins (i.e. Rev, Gag). However, in large part it is host factors that play essential roles in the movement of HIV-1 RNA from the site of transcription to its ultimate encapsidation into new virions. Identifying these factors and their mechanism of action provides not only important insights into HIV-1 molecular biology but also that of the cell machinery itself. In this review, we highlight the viral and host factors regulating the splicing, polyadenylation, transport, and translation of HIV-1 RNA. The observations made underline the multiple fate decisions that must be made at each stage of the viral RNA metabolic pathway and highlight potential new avenues for controlling HIV-1 replication. PMID- 18508614 TI - Hyaluronan and hyaluronidase in genitourinary tumors. AB - Genitourinary cancers are the most frequently diagnosed cancers in men and the fifth most common in women. Management of disease through accurate and cost effective early diagnostic markers, as well as identification of valid prognostic indicators, has contributed significantly to improved treatment outcomes. In this review, we will discuss the function, regulation and clinical utility of hyaluronan (HA), genes encoding its metabolic enzymes and receptors that mediate its cellular effects. Specific HA synthase (HAS) and hyaluronidase (HAase) genes encode the enzymes that produce HA polymers and oligosaccharides, respectively. Differential effects of these enzymes in progression of genitourinary tumors are determined by the relative balance between HAS and HAase levels, as well as the distribution of receptors. The genes are regulated in a complex fashion at the transcriptional and post-translational levels, but also by epigenetic events, alternative mRNA splicing, and subcellular localization. Importantly, the major tumor-derived HAase enzyme, HYAL-1, either alone or together with HA, is an accurate diagnostic and prognostic marker for genitourinary tumors. PMID- 18508617 TI - Clinical review of deep brain stimulation and its effects on limbic basal ganglia circuitry. AB - This paper aims to provide an overview of factors that may contribute to cognitive and mood alterations following DBS (and lesion therapy). A PubMed search based on studies in the English-language was undertaken, and included all publications on the topics of mood and surgery for movement disorders. Information was collected on pre-operative and post-operative characteristics of each study, and study methodology was examined. One-hundred and forty published articles were selected and reviewed for mood and behavior changes following neurosurgery for movement disorders. There were a variety of mood and cognitive changes associated with DBS (and lesion therapy). These alterations of behavior and cognition were seen in all targets but not frequently reported with thalamic DBS. However methodological limitations, small sample sizes, lack of control groups, and the heterogeneity of data reported underscore why interpretation and comparison of limbic effects of DBS remains challenging. The collection and reporting of more standardized minimal data sets will allow for future comparisons, and improve the power required to answer many of the questions raised in this review. PMID- 18508618 TI - In vitro assays of molecular motors--impact of motor-surface interactions. AB - In many types of biophysical studies of both single molecules and ensembles of molecular motors the motors are adsorbed to artificial surfaces. Some of the most important assay systems of this type (in vitro motility assays and related single molecule techniques) will be briefly described together with an account of breakthroughs in the understanding of actomyosin function that have resulted from their use. A poorly characterized, but potentially important, entity in these studies is the mechanism of motor adsorption to surfaces and the effects of motor surface interactions on experimental results. A better understanding of these phenomena is also important for the development of commercially viable nanotechnological applications powered by molecular motors. Here, we will consider several aspects of motor surface interactions with a particular focus on heavy meromyosin (HMM) from skeletal muscle. These aspects will be related to heavy meromyosin structure and relevant parts of the vast literature on protein surface interactions for non-motor proteins. An overview of methods for studying motor-surface interactions will also be given. The information is used as a basis for further development of a model for HMM-surface interactions and is discussed in relation to experiments where nanopatterning has been employed for in vitro reconstruction of actomyosin order. The challenges and potentials of this approach in biophysical studies, compared to the use of self-assembly of biological components into supramolecular protein aggregates (e.g. myosin filaments) will be considered. Finally, this review will consider the implications for further developments of motor-powered lab-on-a-chip devices. PMID- 18508619 TI - DNA-directed assembly of protein microarrays. AB - Microarray technology has made it possible to simultaneously study the abundance, interactions, and functions of potentially tens of thousands of biological molecules. From its earliest use in DNA microarrays, where only nucleic acids were captured and detected on the arrays, applications of microarrays now extend to those involving biomolecules such as antibodies, proteins, peptides, and carbohydrates. In contrast to the relative robustness of DNA microarrays, the use of such chemically diverse biomolecules on microarray formats presents many challenges in their fabrication as well as application. Among the many methods that have been proposed to overcome these challenges, DNA-directed assembly (DDA) has emerged as a promising strategy for the high sensitivity and multiplexed capture and detection of various analytes. In this review, we explore the challenges faced during the design, fabrication, and utilization of protein microarrays and highlight how DDA strategies, together with other recent advances in the field, are accelerating the development of platforms available for protein microarray applications. PMID- 18508620 TI - Toll-like receptor signaling in the ischemic heart. AB - Toll-like receptors (TLRs) have been identified as primary innate immune receptors. They distinguish among different patterns of pathogens and rapidly activate an innate immune response. However, TLRs can also be stimulated by host derived molecules. TLRs are expressed in the cardiovascular system and could thus be a key link between cardiovascular diseases and the immune system. Increasing evidence suggests that TLR signaling promotes injury in the heart after ischemia, ischemia/reperfusion or hypertrophic stimuli. Herein we review the experimental and clinical evidence for the involvement of TLR and its downstream signaling molecules in cardiac ischemic injury. PMID- 18508621 TI - Cystatin C and cathepsins in cardiovascular disease. AB - Cystatin C and cathepsins could play a role in almost all processes involved in atherosclerotic lesion formation by their degradation of extracellular matrix proteins and apolipoprotein B100, the protein moiety of LDL. Several cysteine cathepsins are upregulated in human lesions accompanied by a decrease in cystatin C, the major inhibitor of cysteine cathepsins. Recent research show that atherosclerotic mice deficient in cystatin C display increased elastic lamina degradation as well as larger plaque formation. Cathepsin S- and K-deficient atherosclerotic mice, on the other hand, both have less atherosclerosis, where cathepsin S-/- mice exhibited fewer plaque ruptures and cathepsin K-/- larger foam cells than control mice. This article reviews possible roles of cystatin C and cathepsins in different processes and stages of the atherosclerotic disease. PMID- 18508622 TI - Expression of cytochrome P450 in non-small cell lung cancer. AB - Lung cancer accounts for most of cancer-related deaths in both men and women. Lung cancer is also associated with cigarette smoking that exposes the individual to carcinogenic chemicals. Normally, CYP enzymes (cytochrome P450s) metabolize carcinogens to inactive derivatives, however, occasionally the action of CYP enzymes leads to development of more potent carcinogens. In addition to the metabolism of carcinogenic compounds, CYP enzymes are also involved in the activation and/or inactivation of agents, which are used in the treatment of lung cancer. Therefore, the local level of CYP enzymes in lung cancer and surrounding tissues could be an important determinant in the efficacy of anticancer drugs. Furthermore, the expression of CYP19 (aromatase), estrogen synthesis P450, was found in more than 80 percent of non-small cell lung cancers. Lung cancer was also found to frequently express CYP24A1 that converts 1 alpha, 25 dihydroxyvitamin D3 to its inactive 24-hydroxylated derivatives. The understanding of the local expression of CYP enzymes in tumor tissues is important in the development of better treatment for lung cancer and a standardized treatment, tailor-made, for individual patients. PMID- 18508623 TI - Prostaglandins in labor--a translational approach. AB - The mechanisms involved in the initiation of human labor are largely unknown. Understanding the molecular pathways is fundamental in both the development of effective therapeutic strategies and intervention to prevent preterm labor. Prostaglandins are bioactive lipids and members of the eicosanoids family, derived from arachidonic acid, which act in a paracrine or autocrine manner and function via binding to specific G-protein-coupled receptors, activating intracellular signaling and gene transcription. Prostaglandins have a central role in the maintenance of pregnancy and initiation of labor, with the change from uterine quiescence to a contractile state facilitated by differential expression of prostaglandin receptors within the myometrium and fetal membranes. Clinical evidence for the key role of prostaglandins in human parturition is evident from their successful exploitation as exogenous agents for the induction of labor and the role of prostaglandin synthase inhibitors as a preventative therapy for preterm labor. This review aims to focus on prostaglandin synthesis and metabolism and how differential regulation of prostaglandins and their receptors in gestational tissues interact in the initiation of labor. PMID- 18508624 TI - "Sighted C3H" mice--a tool for analysing the influence of vision on mouse behaviour? AB - It is unclear what role vision plays in guiding mouse behaviour, since the mouse eye is of comparably low optical quality, and mice are considered to rely primarily on other senses. All C3H substrains are homozygous for the Pde6b(rd1) mutation and get blind by weaning age. To study the impact of the Pde6b(rd1) mutation on mouse behaviour and physiology, sighted C3H (C3H.Pde6b+) and normal C3H/HeH mice were phenotyped for different aspects. We confirmed retinal degeneration 1 in C3H/HeH mice, and the presence of a morphologically normal retina as well as visual ability in C3H.Pde6b+ mice. However, C3H.Pde6b+ mice showed an abnormal retinal function in the electroretinogram response, indicating that their vision was not normal as expected. C3H.Pde6b+ mice showed reduced latencies for several behaviours without any further alterations in these behaviours in comparison to C3H/HeH mice, suggesting that visual ability, although impaired, enables earlier usage of the behavioural repertoire in a novel environment, but does not lead to increased activity levels. These results emphasize the importance of comprehensive behavioural and physiological phenotyping. PMID- 18508625 TI - Pancreatic acinar-to-beta cell transdifferentiation in vitro. AB - Although accumulating evidence indicates that proliferation of pre-existing beta cells is the major mechanism of the maintenance of postnatal beta-cell mass, new beta-cells can be generated from non-beta-cells under certain conditions in vitro. We have recently shown directly by Cre/loxP-based cell lineage tracing that adult mouse pancreatic acinar cells can be transdifferentiated into insulin secreting cells in vitro. These newly made cells secrete insulin in response to glucose and other secretagogues, but their secretory capacity is still low compared to that of native beta-cells. To improve the efficiency of generation of insulin-secreting cells from non-beta cells, it is critical to understand the molecular mechanism of such transdifferentiation. Since pancreatic acinar cells are the most abundant cell type in the pancreas, their utilization as a source of surrogate beta-cells is an intriguing approach to cell replacement therapy for type 1 diabetes. This review focuses on current knowledge of the regeneration of pancreatic beta-cells and transdifferentiation of pancreatic acinar-cells into insulin-secreting cells. PMID- 18508626 TI - Unreplicated DNA in mitosis precludes condensin binding and chromosome condensation in S. cerevisiae. AB - Condensin is the core activity responsible for chromosome condensation in mitosis. In the yeast S. cerevisiae, condensin binding is enriched at the regions where DNA replication terminates. Therefore, we investigated whether DNA replication completion determines the condensin-binding proficiency of chromatin. In order to fulfill putative mitotic requirements for condensin activity we analyzed chromosome condensation and condensin binding to unreplicated chromosomes in mitosis. For this purpose we used pGAL:CDC6 cdc15-ts cells that are known to enter mitosis without DNA replication if CDC6 transcription is repressed prior to S-phase. Both the condensation of nucleolar chromatin and proper condensin targeting to rDNA sites failed when unreplicated chromosomes were driven in mitosis. We propose that the DNA replication results in structural and/or biochemical changes to replicated chromatin, which are required for two phase condensin binding and proper chromosome condensation. PMID- 18508627 TI - The tumor antigen EpCAM: tetraspanins and the tight junction protein claudin-7, new partners, new functions. AB - The cell-cell adhesion molecule EpCAM/CD326 has been one of the first tumor associated antigens and has soon received attention as an antibody target in cancer therapy. However, only recently, progress has been achieved in disclosing the array of functional activities of EpCAM and the underlying molecular mechanisms. This review will particularly focus on cooperative activity of EpCAM with two classes of transmembrane molecules, tetraspanins and claudins. EpCAM can associate with claudin-7 and the tetraspanins CD9 and CO-029. We propose that complex formation of EpCAM with tetraspanins and claudins does not only interfere with EpCAM-mediated homotypic cell-cell adhesion, but importantly, is also associated with a gain of function, like induction of apoptosis resistance. PMID- 18508628 TI - MSK activation and physiological roles. AB - Mitogen and stress activated protein kinase (MSK) 1 and 2 are nuclear serine/threonine protein kinases that are activated in vivo downstream of either the ERK1/2 or p38 mitogen activated protein kinase (MAPK) cascades. MSKs contain two kinase domains, an N-terminal kinase domain related to the AGC kinase family, and a C-terminal kinase domain related to the CaMK family. The upstream MAPK phosphorylates the C-terminal domain, which then phosphorylates and activates the N-terminal domain. Once activated, the N-terminal domain phosphorylates substrates. MSKs do not have a precisely defined substrate consensus sequence, however the do have a preference for a basic cluster prior to the phosphorylated residue. In cells MSKs phosphorylate several substrates including CREB, NFkB, HMGN1 and histone H3. The major role of MSKs appear to be in the regulation of immediate early (IE) genes, and consistent with this the transcription of several CRE dependent IE genes is compromised in MSK knockouts. The physiological roles of MSKs still remain to be completely determined, however recent work has suggested a role for MSKs in neuronal synaptic plasticity and in regulating cytokine production in the innate immune system. PMID- 18508629 TI - HPV-16 RNA processing. AB - To understand human papillomavirus type 16 (HPV-16) gene regulation, it is necessary to understand HPV-16 RNA processing. HPV-16 encodes multiple 5'- and 3' splice sites and two polyadenylation signals pAE and pAL (Figure 1). The major 3' splice site on the HPV-16 genome (SA3358) is used for generation of E6, E7, E4, L1 and L2 mRNAs. It encodes a suboptimal splice signal but is under control of a strong enhancer that renders SA3358 one of the most efficiently used splice sites on the HPV-16 genome. Thereby SA3358 indirectly blocks HPV-16 late gene expression. The early polyA signal is also under control of the early UTR sequence and multiple RNA elements in the L2 coding region that interact with hnRNP H. The two splice sites SD3632 and SA5639 are used exclusively by late mRNAs and are under control of multiple splicing silencer elements. The silencers at SA5639 are located in the L1 coding region and interact with hnRNP A1. So far, only polypyrimidine tract binding protein (PTB) has been shown to induce late gene expression. PMID- 18508630 TI - Tracking the mechanisms of deep brain stimulation for neuropsychiatric disorders. AB - Deep brain stimulation (DBS) has recently emerged as a potential treatment for medically intractable neuropsychiatric disorders. Pilot clinical studies with encouraging results have been performed with DBS of the ventral anterior internal capsule (VAIC) and subgenual cingulate white matter (Cg25WM) for the treatment of obsessive-compulsive disorder and depression. However, little is known about the underlying response of individual neurons, or the networks they are connected to, when DBS is applied to the VAIC or Cg25WM. This review summarizes current understanding of the response of axons to DBS, and discusses the general brain network architectures thought to underlie neuropsychiatric disorders. We also employ diffusion tensor imaging tractography to better understand the axonal trajectories surrounding DBS electrodes implanted in the VAIC or Cg25WM. Finally, we attempt to reconcile various data sets by presenting generalized hypotheses on potential therapeutic mechanisms of DBS for neuropsychiatric disease. PMID- 18508631 TI - BRCT domains: phosphopeptide binding and signaling modules. AB - The BRCA1 C-terminus (BRCT) domains are essential for the tumor suppressor function of BRCA1, and have been found in a variety of proteins from bacteria to men. Recent studies demonstrate that the BRCT domain constitutes a novel phosphopeptide binding region. In this review we seek to discuss the recent biochemical and structural data that have helped elucidate the molecular basis of BRCT domain function and BRCT-mediated interactions, with special emphasis on the role of phospho-specific interactions in key networks that regulate DNA repair. Finally we offer predictions on additional phospho-interacting BRCT domains and potential in vivo binding sites for several BRCT domains. PMID- 18508632 TI - Epstein-Barr virus vaccine development: a lytic and latent protein cocktail. AB - Epstein-Barr Virus (EBV) is the causative agent of acute infectious mononucleosis and associates with malignancies such as Burkitt lymphoma, nasopharyngeal carcinoma, and non-Hodgkin's lymphoma. Additionally, EBV is responsible for B lymphoproliferative disease in the context of HIV-infection, genetic immunodeficiencies and organ/stem-cell transplantation. Here we discuss past and current efforts to design an EBV vaccine. We further describe preliminary studies of a novel cocktail vaccine expressing both lytic and latent EBV proteins. Specifically, a tetrameric vaccinia virus (VV) -based vaccine was formulated to express the EBV lytic proteins gp350 and gp110, and the latent proteins EBNA-2 and EBNA-3C. In a proof-of-concept study, mice were vaccinated with the individual or mixed VV. Each of the passenger genes was expressed in vivo at levels sufficient to elicit binding antibody responses. Neutralizing gp350 specific antibodies were also elicited, as were EBV-specific T-cell responses, following inoculation of mice with the single or mixed VV. Results encourage further development of the cocktail vaccine strategy as a potentially powerful weapon against EBV infection and disease in humans. PMID- 18508633 TI - Cytogenetic biomarkers for human cancer. AB - Human cancer cytogenetics is the study of chromosomal rearrangements and numerical abnormalities in malignant tissue. Since the 1960s and the discovery of the Philadelphia chromosome, hundreds of common and characteristic chromosomal aberrations have been observed in various neoplasias. Because these cytogenetic aberrations provide diagnostic, prognostic, and treatment-related information for the associated cancers, they are considered biomarkers for disease. Here we describe many of the best-known chromosome rearrangements and variant rearrangements in hematologic disease and solid tumors, indicate the genes and underlying molecular mechanisms known to be involved in development and progression of disease, and describe the newer molecular cytogenetic technologies and how they are currently being used in cancer diagnostics. We also highlight many important pit-falls in obtaining, transporting, and analyzing neoplastic samples which can compromise cytogenetic studies and preclude its use as a diagnostic tool. PMID- 18508634 TI - Liver receptor homolog-1, an emerging metabolic modulator. AB - The liver receptor homolog-1 (LRH-1; NR5A2) belongs to nuclear hormone receptor superfamily, and is expressed mainly in the liver, intestine, exocrine pancreas, and ovary. It binds DNA as a monomer, and is best known as a regulator of hepatic expression of the key bile acid biosynthetic enzyme cholesterol 7alpha hydroxylase (Cyp7A1). It is also expressed in embryonic stem cells and the initial stages of embryonic development, and the very early lethality of LRH-1 knockout mice highlights its essential developmental role. Recent crystal structures of LRH-1 and its closest relative steroidogenic factor-1 (SF-1; NR5A1) identified phospholipids as potential ligands. This intriguing discovery raises the possibility of an unexpected new class of nuclear receptor signaling molecules, but the broader functional roles of LRH-1 and these new ligands remain to be established. PMID- 18508636 TI - Mechanisms of and obstacles to iron cardiomyopathy in thalassemia. AB - Thalassemia is anemia of variable severity, arising from mutations of genes encoding the hemoglobin alpha and beta chains. Severe thalassemia is associated with iron overload, tissue lesions, and high risk for cardiovascular complications, and iron-mediated cardiomyopathy is the main cause of death in this condition. Thalassemia major (TM) patients exhibit cardiovascular abnormalities consistent with chronic anemia; these include enlargement of the ventricular chambers, increased cardiac output, and reduced total vascular resistance. Cardiac iron overload in TM patients due to long-term transfusion can cause further chamber dilation, decreased contractility, and arrhythmia. Paradoxically, many such patients remain asymptomatic until decompensation occurs. For decades, magnetic resonance imaging and echocardiography have been performed to detect advanced cardiac dysfunction; however, reliable evaluation tools for the early detection of cardiac abnormalities are currently in demand. This article reviews the mechanisms underlying the development of heart disease in thalassemia and strategies for therapeutic intervention in TM patients with congestive heart failure. PMID- 18508635 TI - Sensory roles of neuronal cilia: cilia development, morphogenesis, and function in C. elegans. AB - In the free-living nematode Caenorhabditis elegans, cilia are found on the dendritic endings of sensory neurons. C. elegans cilia are classified as 'primary' or 'sensory' according to the '9+0' axonemal ultrastructure (nine doublet outer microtubules with no central microtubule pair) and lack of motility, characteristics of '9+2' cilia. The C. elegans ciliated nervous system allows the animal to perceive environmental stimuli and make appropriate developmental, physiological, and behavioral decisions. In vertebrates, the biological significance of primary cilia had been largely neglected. Recent findings have placed primary/sensory cilia in the center of cellular signaling and developmental processes. Studies using genetic model organisms such as C. elegans identified the link between ciliary dysfunction and human ciliopathies. Future studies in the worm will address important basic questions regarding ciliary development, morphogenesis, specialization, and signaling functions. PMID- 18508637 TI - Phospholamban phosphorylation by CaMKII under pathophysiological conditions. AB - Sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) transports Ca2+ into the SR, decreasing the cytosolic Ca2+ during relaxation and increasing the SR Ca2+ available for contraction. SERCA2a activity is regulated by phosphorylation of another SR protein: Phospholamban (PLN). Dephosphorylated PLN inhibits SERCA2a. Phosphorylation of PLN by either cAMP or cGMP-dependent protein kinase at Ser16 or the Ca2+-calmodulin-dependent protein kinase (CaMKII), at Thr17, relieves this inhibition, increasing SR Ca2+ uptake and SR Ca2+ load. Thus, PLN is a major player in the regulation of myocardial relaxation and contractility. This review will examine the main aspects of the role of CaMKII and Thr17 site of PLN, on different pathophysiological conditions: acidosis, ischemia/reperfusion (I/R) and heart failure (HF). Whereas CaMKII-activation and PLN phosphorylation contribute to the functional recovery during acidosis and stunning, CaMKII results detrimental in the irreversible I/R injury, producing apoptosis and necrosis. Phosphorylation of Thr17 residue of PLN and CaMKII activity vary in the different models of HF. The possible role of these changes in the depressed cardiac function of HF will be discussed. PMID- 18508638 TI - The papillomavirus E2 DNA binding domain. AB - The DNA binding domain of the E2 master regulator from papillomaviruses is the primary effector for most the essential activities controlled by this protein. In this review we focus on the properties of the DNA binding domain of human papillomavirus strain 16 in solution, integrating structure, dynamics, folding, stability, conformational equilibria, and DNA binding mechanism. We discuss the relevance of these processes for the different biological activities, broadening the horizon for antiviral development. In addition, the particular fold of the DNA binding domain only shared with the Epstein-Barr nuclear antigen EBNA1, suggests a link between this unique architecture and the function of viral origin binding proteins of this kind. Finally, the E2 DNA binding domain proved to be an excellent model for addressing fundamental problems of DNA recognition mechanisms and folding of intertwined dimers. PMID- 18508639 TI - Loss of beta-cells with fibrotic islet destruction in type 2 diabetes mellitus. AB - Recent morphologic analyses of human pancreases strongly suggest that a decreased beta-cell mass is observed from the early stages of diabetes and is caused by accelerated apoptosis of the beta-cells. In this article, we propose that fibrotic islet destruction might be one of the important pathogenic mechanisms of the limited capacity of beta-cell proliferation and accelerated apoptosis in diabetic patients. We have found that pancreatic stellate cells (PSCs) are involved in the progression of islet fibrosis in type 2 diabetes. High concentrations of glucose and insulin in islets contribute to PSC activation and proliferation through angiotensin II type 2 (ATII) signaling pathway, although the exact mechanisms remain to be confirmed. Angiotensin-converting enzyme inhibitors attenuate fibrotic islet destructions and that these have some beneficial effects on glucose tolerance. We suggest that PSCs might play a major role for the fibrotic islet destruction in patients with type 2 diabetes, and suppression of PSCs activation and proliferation might be one of the reasonable target to prevent and delay the progression of the type 2 diabetes mellitus. PMID- 18508640 TI - Type I interferon production by nucleic acid-stimulated dendritic cells. AB - Dendritic cells (DC) detect nucleic acid adjuvants through the Toll-like receptor (TLR) or cytosolic sensors. Nucleic acid adjuvants activate DC to produce a variety of soluble factors including proinflammatory cytokines or type I interferons (IFNs). Type I IFN, especially IFN-alpha, induction is a characteristic function of nucleic acid adjuvants and critical for antiviral host defense. Notably, nucleic acids derived from the host as well as from the pathogens can function as immune adjuvants and contributes to the manifestations of autoimmune diseases through the type I IFN induction. Therefore, clarifying the molecular mechanisms for type I IFN induction by nucleic acids should contribute to the development of treatment not only for viral infection but also for autoimmune diseases. PMID- 18508641 TI - Maintenance of self-tolerance by apoptotic cell clearance. AB - Innate immune cells are genetically conferred the ability to recognize microorganisms as "non-self", and to induce appropriate immune responses to eliminate them. On the other hand, immune cells should recognize self cells in order to avoid attacking normal tissues. For this purpose, immune cells make use of self-cell corpses. When cells undergo apoptosis, cell corpses are rapidly phagocytosed by phagocytes, such as macrophages and dendritic cells. These phagocytes present self antigens derived from dead cell corpses to induce tolerance. Impairment of apoptotic cell clearance often results in autoimmune disorder. Intravenous injection of dead cell corpses can induce tolerance to cell associated antigens, and this strategy has potential use in the treatment of various autoimmune and inflammatory disorders in human. Injected dead cell corpses are rapidly cleared by phagocytes located in the marginal zone (MZ) of spleen. Among those phagocytes, macrophages play a critical role in the rapid clearance of dead cell corpses, and the subsequent induction of tolerance to cell associated antigens. PMID- 18508642 TI - Specificity of signaling from MAPKs to MAPKAPKs: kinases' tango nuevo. AB - Within the signalling network of mammalian cells, MAPK-activated protein kinases (MAPKAPKs) have been identified downstream to various MAPKs, such as the classical MAPKs, ERK1/2, the stress-activated p38 MAPKs and the atypical MAPKs ERK3/4/5. Here, the current understanding of the specificity of MAPK to MAPKAPK signalling, the underlying mechanisms of protein-protein-interaction and the effects on subcellular localisation are reviewed. It is demonstrated that specificity in this signalling section is maintained by protein domain interactions and by regulated subcellular localisation. These mechanisms enable specific MAPK pathways to act independently via specific MAPKAPKs but also allow different MAPK pathways to cooperate in downstream signalling in a coordinated fashion. PMID- 18508643 TI - Information, probability, and the abundance of the simplest RNA active sites. AB - The abundance of simple but functional RNA sites in random-sequence pools is critical for understanding emergence of RNA functions in nature and in the laboratory today. The complexity of a site is typically measured in terms of information, i.e. the Shannon entropy of the positions in a multiple sequence alignment. However, this calculation can be incorrect by many orders of magnitude. Here we compare several methods for estimating the abundance of RNA active-site patterns in the context of in vitro selection (SELEX), highlighting the strengths and weaknesses of each. We include in these methods a new approach that yields confidence bounds for the exact probability of finding specific kinds of RNA active sites. We show that all of the methods that take modularity into account provide far more accurate estimates of this probability than the informational methods, and that fast approximate methods are suitable for a wide range of RNA motifs. PMID- 18508644 TI - Prediction of protein allergenicity using local description of amino acid sequence. AB - The constant increase in atopic allergy and other hypersensitivity reactions has intensified the need for successful therapeutic approaches. Existing bioinformatic tools for predicting allergenic potential are primarily based on sequence similarity searches along the entire protein sequence and do not address the dual issues of conformational and overlapping B-cell epitope recognition sites. In this study, we report AllerPred, a computational system that is capable of capturing multiple overlapping continuous and discontinuous B-cell epitope binding patterns in allergenic proteins using SVM as its prediction engine. A novel representation of local protein sequence descriptors enables the system to model multiple overlapping continuous and discontinuous B-cell epitope binding patterns within a protein sequence. The model was rigorously trained and tested using 669 IUIS allergens and 1237 non-allergens. Testing results showed that the area under the receiver operating curve (AROC) of SVM models is 0.81 with 76 percent sensitivity at specificity of 76 percent . This approach consistently outperforms existing allergenicity prediction systems using a standardized testing dataset of experimentally validated allergens and non-allergen sequences. PMID- 18508645 TI - T cell responses during allergen-specific immunotherapy of Type I allergy. AB - Although allergen-specific immunotherapy (SIT) has been performed in humans for already a century, the immune mechanisms underlying this treatment are still not entirely solved. Allergen-specific CD4+ T lymphocytes are considered as pivotal for the induction and maintenance of allergic disorders. Consequently, their role for allergy treatment has been--and still is--of great interest. Whereas two decades ago immune deviation, i.e. the switch from the allergic Th2 response to a Th1-like response, was described as the most important alteration induced by SIT, more recently the induction of allergen-specific regulatory T cells producing IL 10 has been considered as a main event causing peripheral T cell tolerance. In view of very recent data indicating that both mechanisms may occur consecutively during allergy treatment this review summarizes the current understanding of the immunological mechanisms involved in allergy vaccination. PMID- 18508646 TI - Endocytic mechanisms and drug discovery in neurodegenerative diseases. AB - Extensive research has been carried out to elucidate the mechanism of neurodegenerative diseases, with special emphasis on lysosomal storage disease (LSD) and Alzheimer's disease (AD). Studies have outlined complicated profiles in both types of disorders for the role of endocytosis in disease pathogenesis and progression. Recent discoveries relating endocytosis to the pathological origin and therapeutic strategy of the diseases have yet to be addressed. In this review, I attempt to demonstrate a comprehensive analysis on the endocytic mechanism of the disease. I propose that LSD could be classified as a late endosomal trafficking disorder. I also highlight that the most critical cellular event in AD--the producing, processing, and trafficking of Abeta42 peptide- dynamically involves the entire endocytic system. I further analyze pipeline drug targets, summarize the development status of current new drugs, share thoughts on potential therapeutic strategies, and reveal that many such strategies are in close association with endocytosis. I emphasize that thoroughly understanding pathologically-relevant endocytic events is the key factor in speeding up discovery and development of novel drugs. PMID- 18508647 TI - Effects of static magnetic fields in biology: role of free radicals. AB - Biological systems can respond to a wide range of static magnetic fields (SMF). Some of these responses seem to be mediated partly through free radical reactions. For example, in magnetic sense and navigation using the geomagnetic field, one of the most promising mechanisms for explaining magnetic compass is "a radical pair mechanism". Biological free radicals are most commonly oxygen or nitrogen based with an unpaired electron, leading to the terms "reactive oxygen species (ROS)" or "reactive nitrogen species (RNS)". When applying SMF to medical treatment, coupling SMF exposure with possible chemotherapy of cancers is a novel fascinating area that SMF could enhance agent-induced ROS production against tumors. In addition, one of the potent mechanisms of SMF effects on hemodynamics and blood pressure has sometimes been linked to nitric oxide pathway. However, health and environmental concerns have been raised because the SMF effects on oxidative stress leading to genetic mutation and apoptosis/necrosis have been found. It seems to take place from free radical generation. PMID- 18508648 TI - Chromatin remodeling and SWI/SNF2 factors in human disease. AB - Chromatin structure and its changes or maintenance throughout developmental checkpoints play indispensable role in organismal homeostasis. Chromatin remodeling factors of the SWI/SNF2 superfamily use ATP hydrolysis to change DNA protein contacts, and their loss-of-function or inappropriate increase leads to distinct human pathologic states. In this review, we focus on the translational view of human pathologic physiology involving SWI/SNF2 superfamily, combining latest finding from basic and clinical research. We discuss in mechanistic terms the consequences resulting from dose alteration of the SWI/SNF2 superfamily ATPases and emphasize the necessity of future human subject-based studies. PMID- 18508649 TI - Enzymology of mammalian NAD metabolism in health and disease. AB - Mounting evidence attests to the paramount importance of the non-redox NAD functions. Indeed, NAD homeostasis is related to the free radicals-mediated production of reactive oxygen species responsible for irreversible cellular damage in infectious disease, diabetes, inflammatory syndromes, neurodegeneration and cancer. Because the cellular redox status depends on both the absolute concentration of pyridine dinucleotides and their respective ratios of oxidized and reduced forms (i.e., NAD/NADH and NADP/NADPH), it is conceivable that an altered regulation of the synthesis and degradation of NAD impairs the cell redox state and likely contributes to the mechanisms underlying the pathogenesis of the above mentioned diseases. Taking into account the recent appearance in the literature of comprehensive reviews covering different aspects of the significance of NAD metabolism, with particular attention to the enzymes involved in NAD cleavage, this monograph includes the most recent results on NAD biosynthesis in mammals and humans. Due to recent findings on nicotinamide riboside as a nutrient, its inclusion under "niacins" is proposed. Here, the enzymes involved in the de novo and reutilization pathways are overviewed. PMID- 18508650 TI - Genetic manipulation of islet cells in autoimmune diabetes: from bench to bedside. AB - Type 1 diabetes (T1D) develops via spontaneous autoimmune destruction of pancreatic beta cells. The immunoprogression and effectors of T1D have been determined using non-obese diabetic (NOD) mouse mode. Transgenic mice overexpressing a variety of transgenes driven by insulin promoter demonstrate that both apoptosis and necrosis lead to islet cells death; furthermore, various immune cells and cytokine effectors are involved in the immunoprogression of T1D. Efficiently halting immune attack in the islet milieu by an effector-specific manner apparently provides the preventive and therapeutic strategies in T1D. Islet transplantation has been reported as an appropriate treatment to accomplish insulin independence and long-term homeostasis of glucose in T1D. However, it is difficult to protect the islet grafts from subsequent immune attack and prolong their survival. In this review, we highlight the transgenic mice that express transgenes restricted in islet cells to depict the complicated interactions of immune cells in inflammatory islets, to investigate the protective efficacy of some immunomodulatory genes, and to develop genetically-modified islets tolerant to immune attack that might be used in future clinical application. PMID- 18508651 TI - Regeneration of pancreatic beta cells. AB - Diabetes mellitus results from inadequate mass of insulin-producing pancreatic beta cells. Type 1 diabetes is characterized by absolute loss of beta cells due to autoimmune-mediated destruction. Type 2 diabetes is characterized by relative deficiency of beta cells due to lack of compensation for insulin resistance. Restoration of deficient beta cell mass by transplantation from exogenous sources or by endogenous regeneration of insulin-producing cells would be therapeutic options. Mature beta cells have an ability to proliferate; however, it has been shown to be difficult to expand adult beta cells in vitro. Alternatively, regeneration of beta cells from embryonic and adult stem cells and pancreatic progenitor cells is an attractive method to restore islet cell mass. With information obtained from the biology of pancreatic development, direct differentiation of stem and progenitor cells toward a pancreatic beta cell phenotype has been tried using various strategies, including forced expression of beta cell-specific transcription factors. Further research is required to understand how endogenous beta cells differentiate and to develop methods to regenerate beta cells for clinically applicable therapies for diabetes. PMID- 18508652 TI - Virus associated innate immunity in liver. AB - Dendritic cells (DCs) sense virus via toll-like receptors (TLR) or retinoic acid inducible gene-I (RIG-I) and evoke a cascade of immune reactions. In myeloid DC (MDC) from hepatitis C virus (HCV)-infected patients, the levels of TLR/RIG-I mediated IFN-beta or TNF-alfa induction are lower than those in uninfected donors, suggesting that their signal transduction in MDC is impaired. Dendritic cells in HCV infection are unresponsive to interferon (IFN)-alfa, thus failing to enhance MHC class-I related chain A/B and subsequent NK cell activation. Alternatively, NK cells from the patients down-regulate DC in the presence of HLA E-expressing hepatocytes by secreting IL-10 and TGF-beta1. Such functional alteration of NK cells in HCV infection is ascribed to the enhanced expression of NKG2A/CD94. Activated NKT cells from the patients produce higher levels of IL-13 but comparable IFN-gamma with those from controls, showing their bias to Th2 type. In pegylated IFN-alfa/ribavirin therapy for chronic hepatitis C, improved DC function is related with successful HCV eradication. In conclusion, cross talks among DCs and innate lymphocytes are critical in shaping immune response against HCV, either spontaneously or therapeutically. PMID- 18508653 TI - An adjuvant role of in situ dendritic cells (DCs) in linking innate and adaptive immunity. AB - Dendritic cells (DCs) work as a natural adjuvant to elicit T cell immunity. Though DCs have been widely used in immunotherapy, little is known about their number and function in patients with cancer or autoimmune disease. In recent studies, antigen has been targeted to DCs through DC-specific receptors, such as DEC205, the mannose receptor and dying cell receptors. However, antigen captured by DCs in the absence of danger signals induces tolerance. Therefore, the duration and/or magnitude of danger signals plays a crucial role in generating an immunogeneic response. Various danger signals, i.e., pathogen-associated molecular pattern (PAMP), damage-associated molecular pattern (DAMP) and the activation of innate lymphocytes, serve as maturation signals for DCs. An immunotherapeutic approach which delivers both maturation signals and antigen to DCs would link the innate and adaptive arms of the immune system for a more effective and global immune response. It is therefore crucial to determine optimal conditions for antigen delivery to DCs in an environment suited to maximally stimulate the immune system. PMID- 18508654 TI - Inflammatory cells in renal allografts. AB - Renal transplants are injured by a variety of diseases and pathways. One important cause for acute and chronic graft failure is rejection. Since the advent of kidney transplantation, it has become apparent that rejection is a cellular and/or antibody mediated inflammatory process with different histologic phenotypes, and clinical degrees of severity. In recent years, the immunohistochemical detection of the complement degradation product C4d has further helped to unravel mechanisms of graft injury. Our brief review of 'renal allograft inflammation' focuses on basic morphologic aspects of rejection. Our goal is to foster the close correlation between 'histologic variants of rejection/inflammation' and molecular signaling cascades including chemokine induced effects. PMID- 18508655 TI - Alpha-galactosylceramide-driven immunotherapy for allergy. AB - We report here that the delivery of both alpha-galactosylceramide (alphaGalCer), a representative ligand for invariant natural killer T (iNKT) cells, and an antigenic polypeptide to marginal zone B cells induces the differentiation of regulatory cells in vivo, and suppresses the secondary antibody responses in mice. Splenic CD21+ CD23- B cells of mice treated with alphaGalCer-liposomes produce IL-10 when co-cultured with iNKT cells, whereas the cells treated with aqueous alphaGalCer fail to do so. Adoptive transfer of the B cells into syngenic mice leads to the expansion of splenic CD11c(low) CD45RB(high) cells, which convert naive CD4+ T cells from RAG2-deficient DO11.10 mice to CD4+ CD25(high) Foxp3+ T cells in the presence of OVA323-339 peptide. Administration of alphaGalCer-OVA-liposomes into OVA-primed mice causes the development of CD4+ CD25(high) Foxp3+ T cells that produce both IL-10 and IFN-gamma, and induced the antigen-specific suppression of the secondary antibody responses when boosted with OVA alone. These results indicate that antigen-containing alphaGalCer liposomes can facilitate the development of tolerogenic antigen-presenting cells and inducible regulatory T cells that are involved in the suppression of immune responses to antigens. PMID- 18508656 TI - T cell regulation of hematopoiesis. AB - It has long been known that thymus-derived lymphocytes (T cells) can produce cytokines that have powerful effects on hematopoiesis. All major classes of T cells--CD4 T helper cells, CD4 regulatory T cells, CD8 T cells, gammadelta T cells and NKT cells--produce a number of cytokines and chemokines that can modulate hematopoiesis. More recent research has shown that specific T helper cell types, such as Th1, Th2 and Th17 cells, with the development of each subset depending on distinct STAT proteins, have the potential to modulate the hematopoietic response in different ways. In a teleological sense, the overall orchestration of the immune response by T helper cells fits with the concept that T helper cells would modulate the production of cells of the innate immune system by regulating hematopoiesis. Here we will review the literature on how T cell subsets regulate hematopoietic cell differentiation, and discuss how this regulation may complement the specific function of the T cell type. PMID- 18508658 TI - Cytokine regulation networks in the cancer microenvironment. AB - During carcinoma formation, cancer cells release various cytokines and growth factors into their surroundings and recruit and reprogram many other types of cells in order to establish a tumor microenvironment. Consequently, the tumor tissues almost always contain a large number of endothelial cells, fibroblasts, and infiltrating inflammatory cells that in turn produce a variety of cytokines. The cytokines produced by these cells have been posited as key factors in modulating immune response either against or in favor of tumorigenesis in the microenvironment. The interactions that take place between immune and cancer cells are complex, involving multiple cascades of cytokines, chemokines, and/or growth factors. In this review, we address the essential pro- and anti tumorigenic roles of cytokines in the tumor microenvironment. As the interaction of cytokines, growth factors, and cancer cells forms a comprehensive network at the tumor site that is then responsible for the overall progression or rejection of the tumor, the current review links the microenvironment-derived cytokines and growth factors to a number of different kinds of human carcinogenesis models. Multifunctional cytokines, extracellular matrix mediators, and regulatory cytokines in the cancer environment are all shown to be key factors in the different cancer immune-editing systems. The characterization of cytokine networks in various types of cancer cells may yield important information for understanding the immune-related mechanisms of cancer development, and this knowledge may have subsequent application in cancer immunotherapy. PMID- 18508657 TI - The different functions of Stat5 and chromatin alteration through Stat5 proteins. AB - Stat5 proteins modulate gene transcription upon cytokine- and growth factor action. Stat5a and Stat5b proteins alone are weak activators of transcription. They can modify chromatin organization through oligomerization and they act predominantly in co-operation and interaction with other proteins. The conservative view of exclusively nuclear functions of Stat5 was challenged by the observation of additional Stat5 effects in the cytoplasm, resulting in activation of the PI3K-Akt pathway. We summarize biological consequences of mutations in conserved domains of Stat5 or of deletions in the N- or C-terminal domains with impact on target gene transcription. Formation of higher-order oligomers is dramatically changed upon amino- or carboxyterminal deletions in Stat5 proteins. Mutations in or deletion of the Stat5 N-terminus leads to diminished leukemogenic potential of oncogenic Stat5, probably due to the inability to form Stat5 tetramers. The Stat5 N-terminal domain prevents persistent activation and can act as a DNA-docking platform for the glucocorticoid receptor (GR). The corresponding protocols should facilitate follow-up studies on Stat5 proteins and their contribution to normal physiological versus pathological processes through differential chromatin binding. PMID- 18508659 TI - Variation of the response to the optokinetic drum among various strains of mice. AB - The optokinetic drum has become an appropriate tool to examine visual properties of mice. We performed baseline measurements using mice of the inbred strains C3H, C57BL/6, BALB/c, JF1, 129 and DBA/2 at the age of 8-15 weeks. Each individual C57BL/6, 129 and JF1 mouse was reliably identified as non-affected in vision by determining head-tracking responses. C3H mice were used as negative control because of their inherited retinal degeneration; as expected, they did not respond to the moving stripe pattern. Surprisingly, BALB/c and DBA/2 mice showed the same result. Electroretinography, funduscopy and histology of BALB/c mice did not reveal any abnormality concerning the structure or function of the retina and the remaining eye. Therefore, it might be assumed that BALB/c mice suffer from disturbances of the central visual system. Preliminary results from linkage analysis of the non-responding phenotype in the BALB/c mice indicate a recessive, monogenic mode of inheritance; the causative gene is located on chromosome 7, but significantly different from the albino locus. In conclusion, C57BL/6, 129 and JF1 represent appropriate inbred strains for high throughput screenings with the optokinetic drum. PMID- 18508660 TI - Bile acids and gene regulation: from nuclear receptors to chromatin. AB - Transcription regulation by bile acids is far more complicated than it appeared at first when several groups began their investigations in the early '90. It has become clear now that bile acids regulate the transcription of genes involved in bile acid synthesis, transport and other metabolic pathways via multiple mechanisms that involve transcription factors, nuclear receptors, coregulators, chromatin and the related modifying enzyme complexes. At a first look this might seem surprising but if one considers the physical-chemical properties of these molecules it should be evident that, due to their detergent properties, bile acids may be harmful if they reach high concentrations in the liver and intestine. Therefore, living organisms have developed biochemical mechanisms that finely tune the concentration of bile acids according to the body needs and in response to environmental challenges. In this review, we will discuss the most recent evidences on the mechanisms through which bile acids regulate gene transcription, including the function of nuclear receptors and emphasizing the emerging role of chromatin and the associated modifying enzymes. PMID- 18508661 TI - Control of atherosclerotic plaque vulnerability: insights from transgenic mice. AB - Atherosclerosis is a complex, progressive disease of the large systemic arteries. This multi-factorial disease is characterized by accumulation of lipids, cells and extracellular matrix in the vessel wall. The quest to unravel the molecular mechanisms leading to progression of human atherosclerotic plaques has lead to the development of a variety of animal models. Mice are easily amendable to transgenesis and multiple mutant and inbred strains have been generated in which potential regulators are manipulated and subsequently studied for effects on the development and progression of atherosclerosis. The scope of this review is to discuss the relevance, advantages and disadvantages of genetically-engineered mice to investigate mechanisms of plaque vulnerability. Features of human vulnerable lesions, such as large lipid-rich necrotic cores, active inflammation, matrix remodeling and signs of intraplaque hemorrhage are represented in mouse lesions. Here, we will discuss how atherosclerosis is modified by manipulations in apoptosis, lesional lipid metabolism, inflammatory pathways, matrix remodeling and thrombotic pathways in genetically-engineered mice, emphasizing the insights that have been gained from these studies for the control of plaque vulnerability. PMID- 18508662 TI - Early stage cancer cell invasion: signaling, biomarkers and therapeutic targeting. AB - The process of primary cancer invasion of distant organs is multifactorial and multistep. Successful therapeutic management of invasive cancers remains hampered by the multitude of overlapping signaling pathways that initiate and drive cancer cell migration. A crucial early event by which cancer cells switch from localized to invasive states is initiated by the acquisition of autonomous motile properties; a process driven by dynamic assemblies and disassemblies of multiple focal adhesion, cytoskeleton and motor proteins. Several of the protein complexes involved are tightly regulated through posttranslational modifications and intermolecular collisions with partners that occur in a time- and space-dependent manner. These concerted mechanisms are essential for the regulation of cell shape, cell polarity, and cell motility and migration in response to chemotactic signals. This review summarizes the current knowledge in the field and potential clinical implications for molecular pathology and cancer therapeutics. It is not meant to be comprehensive; aspects related to basic signaling are not dealt with extensively in this review. However, the reader is referred to excellent reviews that provide coverage of these topics. PMID- 18508663 TI - Protein lipidation meets proteomics. AB - Protein lipidation is a crucial protein modification involving the attachment of hydrophobic carbon skeletons (C:14-C:60) of various lipid classes--fatty acids, sterols, glycero-, phospho- and glycolipids. The lipid-protein bond frequently (i) involves the N- or C-terminal ends of the target, (ii) requires amide, ether or ester bonds to a small aminoacid, usually Gly or Cys and (iii) depends on proteolytic events. Lipidation results in protein targeting to the membrane, with the protein behaving as a peripheral component and being oriented toward the inside or outside of the cell. The addition of a single lipid is not sufficient for membrane targeting and another signal, often involving additional lipidation, is required. The methods available for predicting lipid modifications to proteins highlight the importance of identifying short protein motifs in a field in which few data are currently available, due to the complex nature of the modification. Full proteome annotation is already feasible with these predictive tools. We show that lipidation may affect 2-4 percent of all proteins in a given proteome and that double-lipidation is widespread. PMID- 18508664 TI - Chemokines: coded messages for T-cell missions. AB - Chemokines and their receptors control leukocyte migration and homing throughout the body in both physiological and pathological conditions. In the context of the adaptive immune system, which requires high efficiency and control, chemokines and chemokine receptors represent a versatile code that orchestrates the "who, where and when" of the immune response by providing the spatio-temporal guidance for T-cell development, priming and effector functions. In addition to their chemotactic properties, chemokines can directly modulate T-cell responses by amplifying signals at the immune synapse and tuning Th1/Th2 polarization. In this review we will discuss the role of chemokines in T-cell biology, following an ideal pilgrimage that spans the key steps of the T-cell life. PMID- 18508665 TI - Confocal mapping of cortical inputs onto identified pyramidal neurons. AB - Using a confocal microscopy protocol, we carried out a microcircuitry investigation of cortical connections in monkey temporal cortex. Inputs were labeled by BDA injections in posterior area TE, and potential postsynaptic pyramidal neuron targets were labeled with EGFP, by injection of retrogradely transported adenovirus. We scored the number and distribution of putative contacts onto dendritic compartments of neurons in different layers. Initial results show that about 50 percent of apical dendrites of layer (L.) 6 neurons receive contacts, as they ascend through L.4 (n=1 brain), but only 30-35 percent of those from L.5 neurons (n=2). Basal dendrites of L.3 neurons also receive few contacts in L.4. This supports the role of layer 4 as an interlaminar relay in association cortex. In addition, our results indicate spatial heterogeneity in the occurrence and number of contacts, possibly due to subtype specificity in target preference. The maximum number of contacts, for a L.2 neuron projecting from anterior to posterior TE, was 29. This approach seems a useful alternative or complement to electron microscopic analyses of long distance connectivity. PMID- 18508666 TI - Stem cells and regeneration in planarians. AB - Understanding stem cells is a major goal of current research because of its potential medical applications. Although great advances have been made, such as the culturing and differentiation of embryonic stem cells and reprogramming of cell fates, many basic questions remain unanswered. Describing the mechanisms underlying regeneration will help to understand the biology of stem cells and therefore to control their behavior. While regeneration is being studied in a variety of models, the planarian is particularly noteworthy. In this model system a fragment as small as 1/279 of the animal can regenerate completely within a few weeks. These animals can also grow and degrow--specifically degenerating certain tissues--according to environmental conditions, thus demonstrating a complete control of their stem cell dynamics. However, one of the most interesting aspects of the planarian model system is the presence of a unique type of stem cell that can differentiate into all cell types found in the organism, including the germ line. This represents a simple, extremely powerful, and accessible stem cell system in which to address a variety of important questions. In the last ten years, molecular, cellular, and bioinformatics tools have been established for use in this model, making it ideally placed for in vivo analysis of stem cells in their natural environment without ethical complications. PMID- 18508667 TI - The renal stem cell system in kidney repair and regeneration. AB - The adult mammalian renal tubular epithelium exists in a relatively quiescent to slowly replicating state, but has great potential for regenerative morphogenesis following severe ischemic or toxic injury. Kidney regeneration and repair occur through three cellular and molecular mechanisms: differentiation of the somatic stem cells, recruitment of circulating stem cells and, more importantly, proliferation/dedifferentiation of mature cells. Dedifferentiation seems to represent a critical step for the recovery of tubular integrity. Dedifferentiation of tubular cells after injury is characterized by the reactivation of a mesenchymal program that is active during nephrogenesis. Epithelial-to-mesenchymal transition (EMT) of renal tubular cells is an extreme manifestation of epithelial cell plasticity. It is now widely recognized as a fundamental process that marks some physiological, such as morphogenesis, as well as pathological events, such as oncogenesis and fibrogenesis. It might be also considered as a key event in the regenerative process of the kidney. Understanding the molecular mechanisms involved in EMT might be useful for designing therapeutic strategies in order to potentiate the innate capacity of the kidney to regenerate. PMID- 18508668 TI - PDX-1 functions as a master factor in the pancreas. AB - Various pancreatic transcription factors are involved in pancreas development and beta-cell differentiation. Among them, pancreatic and duodenal homeobox factor-1 (PDX-1) plays a crucial role in pancreas development and beta-cell differentiation, and maintaining mature beta-cell function. MafA is a recently isolated beta-cell-specific transcription factor and functions as a potent activator of insulin gene transcription. These pancreatic transcription factors also play a crucial role in inducing surrogate beta-cells from non-beta-cells and thus could be therapeutic targets for diabetes. On the other hand, under diabetic conditions, expression and/or activities of PDX-1 and MafA in beta-cells are reduced, which leads to suppression of insulin biosynthesis and secretion. Thus, it is likely that alteration of such transcription factors explains, at least in part, the molecular mechanism for beta-cell glucose toxicity found in diabetes. PMID- 18508669 TI - Systems of pancreatic beta-cells and glucose regulation. AB - We present mathematical models for systems of beta-cells in pancreatic islets. The first topic begins with the effects of noise and coupling strength on bursting action potentials of beta-cells. From the discussion, the regular bursts are produced by a proper amount of noise and coupling strength. Furthermore, the bursting duration and period depend on the cluster size of beta-cells. We also observe the real size of islets mostly consisting of beta-cells and obtain the size distribution of islets. In addition, we derive either log-normal or Weibull distributions of the islet sizes based on recent observation on islet growth. Islets of Langerhans are composed of several endocrine cells which interact with each other. Considering asymmetric and inhibitory interactions of these endocrine cells, we introduce a simple islet model consisting of alpha-, beta-, and delta cells. Finally, a whole feedback model for glucose regulation is constructed, connecting the microscopic bursting mechanism and the macroscopic blood glucose regulation of the body. We analyzed these models via numerical simulations based on in vivo and in vitro experimental data. PMID- 18508670 TI - The regulation of NK cell function and development. AB - NK cells are the lymphocytes that are differentiated from hematopoietic stem cells in bone marrow (BM) through sequential acquisition of functional receptors. They are one of the critical components of innate immune system. The process of NK differentiation involves a variety of factors such as cytokines, membrane factors, and transcription factors in addition to BM microenvironment. NK cells express their own activating and inhibitory receptors. The cytolytic functions of NK cells against target cells are determined by the balance between theses activating and inhibitory receptors. NK cells discriminate self from non-self by MHC class I-binding inhibitory receptor. Once NK cell inhibitory receptors bound to their MHC class I-ligand, the target cells are protected from NK cell-mediated cytotoxicity. The cytolytic effector functions as well as the characteristic surface phenotypes of NK cells are acquired during the differentiation process. NK cells are emerging to apply as therapeutic agents against a variety of cancers by manipulating differentiation processes and intrinsic activities of the NK cell. PMID- 18508671 TI - NK cells interactions with dendritic cells shape innate and adaptive immunity. AB - While natural killer (NK) cells received their name from their ability to mediate spontaneous cytotoxicity, it has recently become clear that they require activation to target most transformed and infected cells. Dendritic cells (DCs) have been shown to mediate NK cell activation during innate immune responses. Surprisingly, this interaction was recently reported to be required to restrict infections by NK cells, and to take place in secondary lymphoid organs. Here we review these recent studies on NK cell interactions with DCs, discuss the molecular mechanisms underlying the cross-talk between these two innate lymphocyte populations, and out-line how DCs and NK cells synergize to enhance innate immunity against microbes and tumors as well as shape the adaptive immune system. Based on this better understanding, we propose that NK cells should be targeted for their protective functions and as an adjuvant during immunotherapy development. PMID- 18508673 TI - Angiogenesis and inflammation in carotid atherosclerosis. AB - Carotid atherosclerosis is a leading cause of cerebrovascular events. The control of cardiovascular risk factors, i.e. tobacco smoking, alcohol abuse, hypertension, dyslipidemia, diabetes and obesity proved to reduce number of fatal and non-fatal strokes but failed to prevent important number of them. Screening for biomarkers in individuals at high risk of symptomatic vascular disease helped to identify some of them. However, as disease is by its nature multifocal, global testing for biomarkers may have limited practical application. New imaging techniques, including direct visualization of artery metabolism, by 18-FDG-PET, has brought new tools to study local atherosclerosis progression and individual plaque metabolic activity. Advances in molecular biology helped to identify inflammatory genes and its strong link to angiogenesis. The later, is thought to play a key role in the transformation to unstable plaque. Studies of the complex role that plays angiogenesis in plaque development will help in future to design effective therapies addressed at the individual cell level. The purpose of the review is to bring new insights into complicated pathophysiology of carotid atherosclerosis. PMID- 18508672 TI - Viral infection and human disease--insights from minimotifs. AB - Short functional peptide motifs cooperate in many molecular functions including protein interactions, protein trafficking, and posttranslational modifications. Viruses exploit these motifs as a principal mechanism for hijacking cells and many motifs are necessary for the viral life-cycle. A virus can accommodate many short motifs in its small genome size providing a plethora of ways for the virus to acquire host molecular machinery. Host enzymes that act on motifs such as kinases, proteases, and lipidation enzymes, as well as protein interaction domains, are commonly mutated in human disease, suggesting that the short peptide motif targets of these enzymes may also be mutated in disease; however, this is not observed. How can we explain why viruses have evolved to be so dependent on motifs, yet these motifs, in general do not seem to be as necessary for human viability? We propose that short motifs are used at the system level. This system architecture allows viruses to exploit a motif, whereas the viability of the host is not affected by mutation of a single motif. PMID- 18508674 TI - Overexpression of hypoxia/inflammatory markers in atherosclerotic carotid plaques. AB - Hypoxia, angiogenesis and inflammation leads to plaque progression and remodelling and may significantly contribute towards plaque rupture and subsequent cerebrovascular events. Our aim was to study, markers of hypoxia and inflammation previously identified by microarray analysis, in atherosclerotic carotid arteries with low to moderate stenosis. We hoped to describe different cellular populations expressing the studied markers. The location of selected inflammatory molecules obtained as vascular transplants from organ donors were analysed by immunohistochemistry with monoclonal and polyclonal antibodies. Paraffin-embedded sections were cut and probed with antibodies recognizing active B and T-lymphocytes (CD30), hypoxia-inducible factor-1alpha, endoglin (CD105), Interleukin-6 and C-reactive protein. We observed a notable overexpression of HIF 1alpha in inflammatory and hypoxic areas of carotid arteries in all types of lesions from type II-V taken from the patients with carotid stenosis less than 50%. This suggests that HIF-1alpha may have a putative role in atherosclerosis progression and angiogenesis. Dynamic changes in the non-occluding plaques may explain some of the clinical events in patients with low to moderate carotid stenosis. PMID- 18508675 TI - The normal cellular prion protein and its possible role in angiogenesis. AB - Cellular Prion Protein (PrPc) is a ubiquitous glycoprotein present on the surface of endothelial cells. Resting vascular endothelial cells show minimum expression of PrPc and can constitutively release PrPc. PrPc participates in cell survival, differentiation and angiogenesis. During development, neonatal brain endothelial cells transiently express PrPc. Our group recently reported upregulation of PrPc in microvessels from ischemic brain regions in stroke patients. Ischemia/hypoxia induces PrPc expression through the activation of extracellular signal-regulated kinase (ERK). All these data suggest that PrPc plays an important role in angiogenic responses. In addition, PrPc participates in cellular function in the central nervous system, since PrPc is also highly expressed in neurons. PrPc binds copper, suggesting a role in copper metabolism. PrPc also protects cells against oxidative stress and it seems to be involved in neuroprotection. Several studies have demonstrated that PrPc prevents cells from apoptosis and subsequent tissue damage. Moreover, PrPc plays an important role in the immune response. Here, we review the multiple functions of PrPc with a special attention to its recently reported role in angiogenesis. PMID- 18508676 TI - Of alphas and betas: distinct and overlapping functions of STAT3 isoforms. AB - STAT3 is a pleiotropic factor activated by many different signals including cytokines, growth factors and oncogenes. It is involved in a striking number of functions and can activate distinct repertoires of genes in different contexts. Like other STAT factors, STAT3 exists in two isoforms generated by alternative splicing, the full length STAT3alpha and the truncated STAT3beta, generally thought to act as a dominant negative factor. However, STAT3beta is not transcriptionally inactive and is able to both activate and repress genes depending on cellular environment. These unique properties of the STAT3beta isoform may contribute to the extraordinary functional complexity of STAT3 physiological and pathological actions, revealed by conditional mutagenesis studies and not yet fully understood. With this in mind, we try here to summarize what is known about the structure and function of the alpha and beta STAT3 isoforms, both in vitro and in vivo. In addition, we report unpublished data describing the phenotype of mice where the STAT3alpha isoform was specifically ablated. PMID- 18508677 TI - Dynamin and perforin are associated with neovascularisation in advanced carotid plaques. AB - Intimal plaque neovascularization is associated with the development of symptomatic disease and thrombosis, with new 'leaky' fragile microvessels prone to haemorrhage. Perforin or pore forming protein is involved in vascular cell death by forming pores in target cells. Enzymes, in particular, granzyme B are secreted by immune infiltrates present in inflammatory plaque regions and have been shown to induce endothelial cell apoptosis. Similarly, dynamin-2 is a GTPase which mediates oxidised low density lipoprotein-induced apoptosis and is also required for granzyme B-mediated exocytosis and apoptosis. Our pilot studies identified increased expression of these proteins in complicated atherosclerotic plaques. Here we demonstrate by immunohistochemistry that both proteins are over expressed in angiogenic regions of complicated carotid plaques. Dynamin-2 was extensively localised around microvessels and in immune infiltrating cells whilst perforin was localised in immune infiltrating cells, endothelial cells and smooth muscle cells. Over-expression of these proteins may contribute to plaque destabilisation by increasing cellular apoptosis in vulnerable atherosclerotic plaques. PMID- 18508678 TI - Regulation of epithelial and endothelial junctions by PAR proteins. AB - The organization of tissues depends on intercellular junctions that connect individual cells to each other. In sheets of epithelial cells the junctions contain different components like adherens junctions or tight junctions in an asymmetric distribution along the cell-cell contacts. Tight junctions are located at the most apical region of cell junctions, act as a regulatable barrier for small solutes, and separate the apical membrane domain from the basolateral membrane domain. For a long time, the mechanisms that underly the formation of tight junctions and the development of apico-basal membrane polarity in epithelial cells have been poorly understood. Recently, strong evidence has been provided which implicates a conserved set of cell polarity proteins--the PAR proteins--in this process. Here we discuss the mechanisms by which PAR proteins regulate the formation of cell junctions with a special emphasis on vertebrate epithelial cells. PMID- 18508679 TI - Tumor microenvironment and angiogenesis. AB - The tumor microenvironment is a mixture of extracellular matrix molecules, tumor cells, endothelial cells, fibroblasts and immune cells. Tumor growth and metastasis formation are dependent on the growth of blood vessels into the tumor mass. The tumor microenvironment contributes to this pathological angiogenic process. The extracellular matrix and basement membranes are a source for endogenous angiogenesis inhibitors, such as endostatin. On the other hand, many extracellular matrix molecules can promote angiogenesis by stabilizing blood vessels and sequestering pro-angiogenic growth factors. The majority of stromal cells in carcinomas are fibroblasts. Carcinoma-associated fibroblasts show a distinct phenotype from normal fibroblasts. The mechanisms how the tumor associated fibroblasts regulate angiogenesis are not fully known, but they are suggested to be an important source for growth factors and cytokines recruiting endothelial cells. The immune cells, particularly macrophages and neutrophils are another source for angiogenesis-regulating chemokines, growth factors and proteases. Taken together, the tumor microenvironment is a complex unorganized tissue of various cell types and extracellular matrix that can regulate the pathological angiogenic switch. PMID- 18508680 TI - The mitochondrial free radical theory of ageing--where do we stand? AB - Understanding the molecular mechanisms underlying the ageing process may provide the best strategy for addressing the challenges posed by ageing populations worldwide. One theory proposing such molecular mechanisms was formulated 50 years ago. Harman et al. suggested that ageing might be mediated by macromolecular damage through reactions involving reactive oxygen species (ROS). Today, a version of the free radical theory of ageing, focusing on mitochondria as source as well as target of ROS, is one of the most popular theories of ageing. Here we critically review the status of key principles and concepts on which this theory is based. We find that the evidence to date shows that many of the original assumptions are questionable, while on some critical issues further refinements in techniques are required. Even so, it is becoming evident that mitochondria and mtDNA integrity may indeed be crucial determinants of organismal ageing. Implications for the prospect of successful interventions as well as evidence for and against efficacy of current therapeutic approaches are discussed. PMID- 18508681 TI - Understanding eukaryotic linear motifs and their role in cell signaling and regulation. AB - It is now clear that a detailed picture of cell regulation requires a comprehensive understanding of the abundant short protein motifs through which signaling is channeled. The current body of knowledge has slowly accumulated through piecemeal experimental investigation of individual motifs in signaling. Computational methods contributed little to this process. A new generation of bioinformatics tools will aid the future investigation of motifs in regulatory proteins, and the disordered polypeptide regions in which they frequently reside. Allied to high throughput methods such as phosphoproteomics, signaling networks are becoming amenable to experimental deconstruction. In this review, we summarise the current state of linear motif biology, which uses low affinity interactions to create cooperative, combinatorial and highly dynamic regulatory protein complexes. The discrete deterministic properties implicit to these assemblies suggest that models for cell regulatory networks in systems biology should neither be overly dependent on stochastic nor on smooth deterministic approximations. PMID- 18508682 TI - Hemostatic effects of recombinant DisBa-01, a disintegrin from Bothrops alternatus. AB - A monomeric RGD-disintegrin was recently identified from a cDNA library from the venom gland of Bothrops alternatus. The corresponding 12 kDa-recombinant protein, DisBa-01, specifically interacted with alpha(v)beta3 integrin and displayed potent anti-metastatic and anti-angiogenic properties. Here, the interaction of DisBa-01 with platelet alphaIIb beta3 integrin and its effects on hemostasis and thrombosis were investigated. DisBa-01 bound to Chinese Hamster Ovary (CHO) cells expressing beta3 or alphaIIb beta3 and promoted their adhesion and the adhesion of resting platelets onto glass coverslips. The disintegrin inhibited the binding of FITC-fibrinogen and FITC-PAC-1 to ADP-stimulated platelets and inhibited ADP-, TRAP- and collagen-induced aggregation of murine, rabbit or human platelets. In a flow chamber assay, DisBa-01 inhibited and reverted platelet adhesion to immobilized fibrinogen. DisBa-01 inhibited the phosphorylation of FAK following platelet activation. The intravenous injection of DisBa-01 in C57Bl6/j mice, prolonged tail bleeding time as well as thrombotic occlusion time in mesenteric venules and arterioles following vessel injury with FeCl3. In conclusion, DisBa 01 antagonizes the platelet alphaIIb beta3 integrin and potently inhibits thrombosis. PMID- 18508683 TI - Disintegrins in health and disease. AB - Few of the proteins isolated and characterized from snake venom have proven to be more chemically diverse, exquisitely specific or promiscuously active than the family known as disintegrins. These small proteins have shown structural homology with hundreds of cell surface molecules from plants and animals other than snakes, and their precise mimicry of native receptor ligands speaks to evolutionary niches related to survival and geographic locale. Over 100 disintegrins have been named and studied, with the most recent efforts into molecular techniques providing significant clues to taxonomic relationships among four different snake families. Investigators have evaluated disintegrin applications in therapies for cancer, asthma, osteopenia and inappropriate angiogenesis. Crystal and NMR studies have confirmed hypotheses regarding ligand receptor interactions while illuminating the complexities of structure-function evidence. Disintegrin chimeras with viruses, microbubbles and fluorescent labels have become useful tools in many investigations. While many disintegrin studies still involve platelets, previously unexplored interactions with glial cancer, T lymphocytes and the bacteria Yersinia have blazed new trails for this field. This review will summarize disintegrin investigations since 2003. PMID- 18508684 TI - Disassembly of endothelial and epithelial junctions during leukocyte transmigration. AB - Leukocyte migration occurs as a response to inflammatory signals and is an efficient host defense mechanism against invading pathogens. This innate defense response includes transendothelial and transepithelial migration of leukocytes to facilitate clearance of inflammatory stimuli. The endothelium lines the vascular system and forms the first barrier for leukocytes as they migrate out of the bloodstream. The epithelium largely separates organs from the external environment and forms a second barrier for leukocytes. These cellular barriers are comprised of complex intercellular junctions of different molecular composition. However, for barrier function to be maintained, these specialized intercellular junctions must not be destroyed during transmigration. Innate immune cells including monocytes, neutrophils and dendritic cells are all capable of a highly regulated transmigration response in order to accomplish their different functions. These cells exploit many common adhesive and signaling cascades to traverse cellular junctions. However, there are unique features of each type of leukocyte and barrier that determine specificity of the response. This review will focus on highlighting the mechanisms that leukocytes exploit to open these junctions. PMID- 18508685 TI - Molecular mechanisms of hyperoxia-induced acute lung injury. AB - Hyperoxia-induced acute lung injury (HALI) is characterized by an influx of inflammatory cells, increased pulmonary permeability, endothelial and epithelial cell death. Reactive oxygen species have been postulated to have a significant role in HALI, in part, by inducing cell death responses regulated by shared common mediators of apoptotic and necrotic pathways. Significant differences exist in the response of the mature and developing lung to HALI. PMID- 18508686 TI - Actin motors that drive formation and disassembly of epithelial apical junctions. AB - Tight junctions (TJ) and adherens junctions (AJ) are the most characteristic morphological features of differentiated epithelia which mediate cell-cell adhesions, establishment of the paracellular barrier and development of apico basal cell polarity. In polarized epithelial cells, TJ and AJ associate with the prominent apical actin cytoskeleton, which is known to stabilize junctional structure and to tighten the epithelial barrier. Furthermore, plasticity of the actin cytoskeleton is thought to be critical for the remodeling of epithelial junctions. Two major molecular motors such as myosin II and actin filament turnover provide driving forces for reorganizations of the actin cytoskeleton. The review focuses on the roles of these actin motors in the establishment, maintenance and disassembly of epithelial TJ and AJ during tissue morphogenesis and in pathology. PMID- 18508687 TI - Exploiting BH3 only protein function for effective cancer therapy. AB - Failure to induce tumour selective, efficient cell killing is a major problem underlying the limitations of conventional cytotoxic chemotherapy. Greater understanding of the downstream death inducing signalling pathways and how they are regulated in the drug resistant setting is crucial for improvement of survival in most solid cancers. Here we review the role played by BH3 only proteins in mediating cell death through BAX and BAK, and how this knowledge has lead to a new generation of targeted agents with promising cancer cell killing efficacy. PMID- 18508688 TI - Role of Lgl/Dlg/Scribble in the regulation of epithelial junction, polarity and growth. AB - Genetic studies in Drosophila have revealed that three tumor suppressors, Discs large (Dlg), Scribble (Scrib) and Lethal giant larvae (Lgl), which localize to the basolateral region of epithelial cells, cooperatively regulate cell polarity, junction formation and cell growth in epithelial cells. Subsequent studies in Drosophila, vertebrates and C. elegans have shown the evolutionary conservation of some of their functions in epithelial cells. Also, these studies revealed the importance of antagonistic interactions between these tumor suppressors and apical polarity regulators such as Crumbs and aPKC for the establishment of apical-basal polarity with organized cell-cell junctions and regulation of cell growth in epithelial cells. PMID- 18508689 TI - Role of Toll like receptor signaling pathway in ischemic coronary artery disease. AB - Inflammatory process plays a fundamental role in ischemic coronary artery disease (CAD) in terms of both the etiology of atherosclerosis and the pathophysiology of CAD. In particular, chronic inflammation plays a key role in coronary artery plaque instability and subsequent occlusive thrombosis. It is therefore important to clarify the mechanism underlying the activation of the immune response in the pathogenesis of CAD. Currently 10 toll-like receptors (TLRs) have been reported in mammalian species, and these appear to recognize distinct pathogen-associated molecular patterns controlling innate immune responses. In recent studies, signaling of two forms of human TLR (TLR2 and TLR4) has been shown to be involved in the pathogenesis of CAD, establishing a key link between the progression of coronary atherosclerosis and immune response to both foreign pathogens and endogenously generated inflammatory ligands. A better understanding of TLR signal may provide a novel therapeutic agent for the treatment of CAD. This review summarizes the relationship between the pathogenesis of ischemic coronary artery disease and the human TLR system. PMID- 18508690 TI - Mammalian ADP-ribosyltransferases and ADP-ribosylhydrolases. AB - ADP-ribosyltransferases (ARTs) and ADP-ribosylhydrolases (ARHs) catalyze opposing reactions, which are termed ADP-ribosylation and de-ADP-ribosylation. ARTs transfer the ADP-ribose unit from NAD (nicotinamide adenine dinucleotide) onto an acceptor, while ARHs release the ADP-ribose from the target. Like protein phosphorylation, ADP-ribosylation is a posttranslational modification regulating protein function. In many cases, ADP-ribosylation inactivates the target protein. Numerous bacterial toxins intoxicate cells by attaching an ADP-ribose moiety to a functionally important amino acid residue, thereby blocking the interaction of the target protein with other proteins. In other cases, ADP-ribosylation activates protein function. On the surface of T cells, ART2.2 ADP-ribosylates the P2X7 purinoceptor on arginine 125, thereby gating the P2X7 ion channel by presenting a ligand to its nucleotide-binding site. ADP-ribosylation is not limited to protein targets and ARTs have been described that ADP-ribosylate DNA, RNA, and small molecules. Mammalian cells express distinct families of ARTs and ARHs. Recently, molecular cloning, site directed mutagenesis and three dimensional structural analyses of prototype mammalian ARTs and ARHs have shed fresh insight into the structure and function of these intriguing enzymes. PMID- 18508691 TI - Structural changes of the zona pellucida during fertilization and embryo development. AB - The zona pellucida (ZP) is a unique extracellular coat surrounding the maturing oocyte, during ovulation, fertilization, and early embryo development. It is formed by three/four glycoproteins. Ultrastructural data obtained with transmission (TEM) and scanning electron microscopy (SEM) were compared with molecular data on the glycoproteins network from ovulation to blastocyst formation. Molecular models are quite different to the morphology obtained with TEM, which shows a microfibrillar architecture, or with SEM, which shows a spongy or smooth surface. The saponin-ruthenium red-osmium tetroxide-thiocarbohydrazide technique allows to show the ZP real microfilamentous structure and the related functional changes. These results support an ultrastructural supramolecular model, more similar and comparable to molecular models related with the glycoprotein network. A detailed mapping of single mammalian ZP proteins and their relationship within the supramolecular architecture of the zona matrix would clearly supply insights into the molecular basis of sperm-egg recognition. Differences in ZP glycoproteins among mammals do not affect structural morphology; further studies are needed to clarify the relationships between ultrastructural and molecular organizations. PMID- 18508692 TI - Intact and cleaved uPAR forms: diagnostic and prognostic value in cancer. AB - The cellular receptor for urokinase, uPAR, localizes its ligand, uPA, and thereby the plasminogen activation, to the cell surface. uPA also cleaves uPAR, liberating the ligand-binding domain I, and thereby inactivates the binding potential of uPAR for both uPA and vitronectin. The uPA-catalyzed cleavage of uPAR is fast on the cell surface, when uPA is bound to a neighboring uPAR molecule. uPAR can be shed from the cell surface. However, the soluble form cannot be cleaved by uPA. Glycolipid-anchored and soluble forms of intact, uPAR(I III), and cleaved receptor, uPAR(II-III) and uPAR(I), have been identified in tissue and body fluids. It is well-established, that the total amount of all uPAR forms is a strong prognostic marker in different types of cancer. Using immunoassays, measuring the individual uPAR forms, has revealed that the cleaved uPAR forms are even stronger prognostic markers and have diagnostic utility. This review will focus on the mechanism of uPAR cleavage and the functional consequences, as well as the clinical applicability of cleaved uPAR forms. PMID- 18508693 TI - New hydroxypyridinone iron-chelators as potential anti-neurodegenerative drugs. AB - The neuroprotective action of a set of new hydroxypyridinone-based (3,4-HP) compounds (A, B and C), which are iron chelators extra-functionalized with a propargylamino group for potential MAO-B inhibition, was evaluated after cell treatment with MPP+ (an in vivo inducer of parkinsonism) and Abeta(1-40) and/or Abeta(1-42) peptides. Our results show that all these compounds improved cell viability in cells treated with MPP+ and Abeta(1-40) peptide or Abeta(1-42) peptide. In order to evaluate the cellular mechanisms underlying the activity of these compounds, we studied their protective role in caspase activation. All compounds tested were able to prevent MPP+ and Brefeldin A induced caspase-2 activation. They also showed quite effective in the inhibition of caspase-4 and caspase-3 activity, an effector caspase in the apoptotic process. Finally, detection of apoptotic-like cell death after cell exposure to MPP+ was also performed by TUNEL assay. Our results demonstrated that all tested compounds prevented DNA fragmentation by decreasing TUNEL positive cells. A, B and C were more effective than DFP (a 3,4-HP iron-chelating agent in clinical use) in MPP+ induced cell death. Therefore, these results evidenced a neuroprotective and antiapoptotic role for the compounds studied. PMID- 18508694 TI - Mechanisms of reorganization of cell-cell junctions in the testis. AB - During spermatogenesis, intercellular junctions are dynamically remodeled to facilitate the translocation of developing germ cells from the basal to the adluminal compartment of the seminiferous epithelium. In particular, at stage VIII of the seminiferous epithelial cycle, timely restructuring of cell junctions localized at the blood-testis barrier and apical ectoplasmic specializations are crucial to allow the entry of preleptotene/leptotene spermatocytes to the adluminal compartment for further development and the release of mature spermatids from the epithelium respectively. In this review, the transcriptional, post-transcriptional and post-translational regulations involved in testicular cell junction restructuring are included and specific areas required further attention are also highlighted. PMID- 18508695 TI - Kinetochore structure and spindle assembly checkpoint signaling in the budding yeast, Saccharomyces cerevisiae. AB - The Spindle Assembly Checkpoint (SAC) delays the onset of anaphase until every chromosome is properly bioriented at the spindle equator. Mutations in SAC genes have been found in tumors and compromised SAC function can increase the incidence of some carcinomas in mice, providing further links between cancer etiology, chromosome segregation defects and aneuploidy. Here we review recent developments in our understanding of SAC control with particular emphasis on the role of the kinetochore, the nature of the tension sensing mechanism and the possibility that the SAC encompasses more than just stabilization of securin and/or cyclin-B via inhibition of the APC/C to delay anaphase initiation. Our primary emphasis is on the SAC in the budding yeast Saccharomyces cerevisiae. However, relevant findings in other cells are also discussed to highlight the generally conserved nature of SAC signaling mechanisms. PMID- 18508696 TI - Chemokines and chemokine receptors in stem cell circulation. AB - Stem cells are rare, pluripotent, self-renewing cells that give rise to all mature cells during development and adult life. Due to their proliferative capabilities and their ability to home and contribute to the regeneration of damage tissue, stem cells can be transformed into established tumors. Stem cells can function as a double-edged sword--they have the ability to circulate and migrate throughout the developing and mature adult organism, which is essential for their normal function; however, transformed stem cells are also endowed with the machinery to metastasize into various organs. Chemokine and chemokine receptors play a critical role in directing the trafficking of these cells. It is therefore evident that understanding the role of chemokines and their receptors in stem cell circulation is critical for the successful use of these cells in therapy for a wide variety of pathological conditions. PMID- 18508697 TI - Dielectrophoretic manipulation of DNA in microelectrode gaps for single-molecule constructs. AB - The construction with biomolecules and their manipulation represent a key step for developing new miniaturized structures. Such micro or nanometer systems promise a variety of novel features. Dielectrophoresis (DEP) is a powerful tool for trapping and orienting individual molecules in microelectrode arrangements, and was demonstrated to be applicable to DNA. This relatively rigid biomolecule could (after defined immobilization) act as template for further modifications and functionalizations, e.g. metallization. Parameters of the DEP process were adapted to the given electrode layout and for trapping a few or even a single DNA strand. Characterization with atomic force microscopy (AFM) extends the standard method of fluorescence imaging by resolving the resulting structures with single molecule resolution. PMID- 18508698 TI - Inhibition of PI3K improves contractility in alpha1-adrenergically stimulated myocardium. AB - Recent studies have demonstrated that phosphoinositide 3-kinases (PI3Ks) play a fundamental role in regulating myocardial contractility. However, even though alpha1-adrenergic receptor stimulation is known to activate PI3Ks, the impact of this pathway on the inotropic effects of alpha1-stimulation is unclear. Isolated rabbit ventricular myocytes were preincubated with the PI3K inhibitor wortmannin (WM, 0.1 micromol/L). The alpha1 agonist phenylephrine (PE, 10 micromol/L) induced a significantly stronger increase in contractility in WM-treated versus control myocytes (Fractional shortening in percent of resting cell length: 6.14+/ 0.33 percent; n=26 versus 4.85+/-0.33 percent; n=26, P less than 0.05). Furthermore, pretreatment with WM significantly increased the positive inotropic effect of PE in intact muscle strips from rabbit hearts. Mechanistically, we demonstrate that in WM-treated myocytes PE increased phospholamban (PLN) phosphorylation and intracellular Ca2+ transients to a significantly greater extent than in control myocytes. In summary, this is the first study to demonstrate that inhibition of PI3K by increasing PLN phosphorylation and Ca2+ transients significantly improves contractility in alpha1-adrenergically stimulated myocardium. This may have clinical implications for the treatment of decreased cardiac function in acute heart failure. PMID- 18508699 TI - Inhibiting dipeptidyl peptidase activity partially ameliorates colitis in mice. AB - New treatment strategies are required for the debilitating inflammatory bowel diseases (IBD), Crohn's Disease and Ulcerative Colitis. DP inhibitors can prolong the bioactivity of the potent intestinotrophic growth factor glucagon-like peptide-2 (GLP-2(1-33)). We investigated whether novel inhibitors of DP activity could modify the course of disease activity in the dextran sulfate sodium (DSS) model of colitis. C57BL/6 mice consumed 2 percent DSS in drinking water for 6 days. Mice were orally gavaged twice daily with 0.9% saline, 10 mg/kg isoleucyl cyano-pyrrolidine (P59/99) or isoleucyl-thiazolidine (P32/98). Assessment of disease severity incorporated a disease activity index (DAI), together with histological assessment of crypt area and depth in the distal colon. DP activity was significantly inhibited at all time points. The DAI was significantly lower in the P59/99 and P32/98 treatment groups compared to saline treatment in all three time courses. Crypt hyperplasia (p<0.05) was observed in the saline group compared to P32/98 treatment at day 9. This preliminary study shows that novel inhibitors of DP activity may provide a new treatment strategy for IBD. PMID- 18508700 TI - Hemocidins in a functional and structural context of human antimicrobial peptides. AB - Hemocidins are a recently discovered group of microbicidal peptides that emerge from heme-binding proteins, especially hemoglobin. Hemocidins have been obtained in vitro after the chemical or enzymatic fragmentation of globin molecules and have also been isolated from biological sources, such as insect guts and the tissues and excretions of the female reproductive tract. This work presents a concise review of contemporary studies concerning antibacterial peptides, especially those derived from humans, and against this broad structural and functional background discusses the properties of hemocidins. PMID- 18508701 TI - Bridging innate and adaptive immunity through gammadelta T-dendritic cell crosstalk. AB - Like Natural Killer cells, gammadelta T cells and Natural Killer T cells display several innate-like features that confer them a broad reactivity against tumors and pathogens. By recognizing stress-induced conserved antigens upregulated a wide array of physiopathological contexts, these lymphoid subsets develop strong and early responses to a broad set of targets. One of the most exciting roles possibly played in vivo by non-conventional T lymphocytes, which exhibit a biased natural memory phenotype, is active regulation of adaptive immune responses through interactions with antigen presenting cells (APCs), such as dendritic cells. Here we will review recent studies reporting functional interactions between gammadelta T cells and APC and a possible involvement of these lymphocytes in bridging innate and adaptative immunity along infections and tumor development. Our discussion will focus on human gammadelta T cells and more specifically on Vgamma9Vdelta2 T cells, a major subset found in human peripheral blood. PMID- 18508702 TI - The basal phylogenetic position of Nanoarchaeum equitans (Nanoarchaeota). AB - Using sequences of ribosomal RNA from organisms belonging exclusively to the Archaea domain and by means of two methods to remove the phylogenetic noise, we investigate the phylogenetic position of Nanoarchaeum equitans. The results obtained are compatible with the hypothesis that N. equitans represents a new phylum within the Archaea domain because the characteristic long branch of N. equitans in phylogenetic trees is conserved even after most of the phylogenetic noise has been removed, thus implying that its rRNA might indeed be singular. However, our analysis is unable to be equally as clear on the phylogenetic position of Methanopyrus kandleri. PMID- 18508703 TI - Isolation and cytotoxicity of low-molecular-weight metabolites of Candida albicans. AB - In this study, the low molecular weight lypophilic metabolites of C. albicans and C. dubliniensis strains produced in a synthetic medium with the addition of fetal calf serum were identified using LC/MS and MS/MS technique and quantified. All strains investigated produce a metabolite with a UV spectra maximum at 224 and 279 nm and minimum at 243 nm. Following comparison with ESI, MS/MS spectral data of a reference compound, the metabolite was identified as 3-indoleethanol (tryptophol). The concentration of extracellular tryptophol in the biosynthesis of C. albicans and C. dubliniensis ranged from 2.45 microg/mL to 191 microg/mL, respectively. Contrary to previously published data, gliotoxin or gliotoxin-like compounds were not detected, and all investigated C. albicans and C. dubliniensis strains have the same metabolite profile. Cytotoxic effects of tryptophol and 3 indolelactic acid (precursor of tryptophol biosynthesis) were cell-line dependent. The EC50 of tryptophol ranged between 2 and 7 mM, with the EC50 of 3 indolelactic acid approximately double (between 4 and 8 mM). Tryptophol exhibited cell-type dependent cytotoxicity in relatively high concentrations, with domination of apoptosis. PMID- 18508704 TI - Proteomics of stress response in Bifidobacterium. AB - Bifidobacteria are commensal microorganisms of the human gastrointestinal tract which are largely being used in functional foods. Some strains are considered as probiotics since they beneficially affect the composition and the metabolic activity of intestinal microbiota, as well as the health status of the host. The lack of genetic tools has hindered the development of functional genomic studies in bifidobacteria, like the identification of molecular mechanisms underlying their survival under different environmental challenges. Some of these experimental obstacles have been successfully overcome with the use of proteomics, a set of techniques that, when applied to microorganisms, are directed to the identification of all the proteins produced by the cells under a given physiological condition. The aim of this review is to discuss and summarize some of the current knowledge of the stress tolerance in bifidobacteria, mainly identified by two-dimensional electrophoresis coupled to mass spectrometry, and compare the most recent proteomic results with the currently available transcriptomic studies. The input and advantages of novel high throughput proteomic techniques are considered as well. PMID- 18508705 TI - Anti-beta-glucan-like immunoprotective candidacidal antiidiotypic antibodies. AB - Mycoses, candidiasis in particular, are relatively common opportunistic infections still characterized by an unacceptable high mortality rate. Furthermore, they are often complicated by resistance or refractoriness to the existing antimicrobial agents. In recent years new effective therapeutic and large-scale preventative strategies have been proposed by exploiting the identification of fungal beta-glucans as target of antifungal agents such as echinocandins, yeast killer toxins and protective antibodies. Anti-beta-glucan antibodies are detectable in animal and human sera. When elicited by glucan-based vaccines they can exert a fungicidal protective activity. Beta-glucan cell wall killer toxin receptors can elicit fungicidal protective antibodies following natural and experimental infections. When used as an immunogen a killer toxin neutralizing monoclonal antibody (beta-glucan-like) is able to elicit a significant anticandidal protection mediated by anti-idiotypic anti-beta-glucan like candidacidal antibodies. Polyclonal, monoclonal and recombinant anti-beta glucan-like antibodies and peptide mimotopes are able to exert an in vitro and/or in vivo microbicidal activity against eukaryotic and prokaryotic killer toxin receptor-bearing pathogenic microorganisms. Implications and perspectives for transphyletic anti-infectious control strategies, as immunoprevention and immunotherapy, are discussed. PMID- 18508706 TI - Overview: Ticks as vectors of pathogens that cause disease in humans and animals. AB - Ticks (Acari: Ixodidae) transmit a wide variety of pathogens to vertebrates including viruses, bacteria, protozoa and helminthes. Tick-borne pathogens are believed to be responsible for more than 100,000 cases of illness in humans throughout the world. Ticks are considered to be second worldwide to mosquitoes as vectors of human diseases, but they are the most important vectors of disease causing pathogens in domestic and wild animals. Infection and development of pathogens in both tick and vertebrate hosts are mediated by molecular mechanisms at the tick-pathogen interface. These mechanisms, involving traits of both ticks and pathogens, include the evolution of common and species-specific characteristics. The molecular characterization of the tick-pathogen interface is rapidly advancing and providing new avenues for the development of novel control strategies for both tick infestations and their associated pathogens. PMID- 18508707 TI - Targeting the tick-pathogen interface for novel control strategies. AB - Ticks are ectoparasites of wild and domestic animals and humans that most notably impact global health by transmitting disease-causing pathogens. While information on the molecular interactions between ticks and pathogens that facilitate pathogen infection, development and transmission is limited, a comprehensive understanding of the tick-pathogen interface would be fundamental toward development of new and novel measures for control of both tick infestations and tick-borne pathogens. Recently, vaccine studies using key tick antigens and characterization of tick gene function by RNA interference (RNAi) have provided new information on genes that impact the tick-pathogen interface. In this review we summarize current research and prospects of tick vaccines and genetic manipulation of ticks targeted to the tick-pathogen interface. The knowledge gained from these collective studies will be fundamental toward understanding of tick-pathogen interactions and for formulation of control methods targeted at both ticks and tick-borne pathogens. Use of these molecular approaches will likely contribute to control measures that will notably reduce tick populations and tick-borne diseases in the future. PMID- 18508708 TI - Structure and stability of amyloid fibrils formed from synthetic beta-peptides. AB - Synthetic peptides capable of self-assembling into amyloid-like fibrillar structures are emerging as novel building blocks for biomaterials. They also serve as simple model systems to study the aggregation process involved in amyloid diseases. In this paper, we probe the structure and stability of fibrillar assemblies formed by two designed peptides P11-I (CH3-CO-Q2RQ5EQ2-NH2) and P11-II (CH3-CO-Q2RFQWQFEQ2-NH2). Our results suggest that the two peptides assemble by fundamentally different mechanisms to structures of different morphologies. Coulombic interactions between charged residues Arginine and Glutamate drive the self-assembly process for peptide P11-I while the hydrophobic effect appears to be the main driving force in the self-assembly of peptide P11 II. PMID- 18508709 TI - DNA repair, insulin signaling and sirtuins: at the crossroads between cancer and aging. AB - For many years organismal aging and cancer were viewed as separate entities. Recent studies however have suggested that these two seemingly disparate biological processes may in fact share common biochemical pathways. One area of emerging convergence involves the intersection of pathways known to mediate DNA repair with pathways previously implicated in insulin signaling. Recent evidence suggests that the sirtuin family of proteins act as central mediators of this molecular crosstalk. The coordination of DNA repair with overall energy balance may be essential for reducing the risk of developing cancer as well as for determining the rate at which we age. This review will summarize our current knowledge on how the maintenance of genomic integrity and insulin signaling intersect, the potential regulation of sirtuins in this crosstalk, and how this coordinated regulation may have important implication for both tumor-free and overall survival. PMID- 18508710 TI - Fibroblasts emerge via epithelial-mesenchymal transition in chronic kidney fibrosis. AB - Our current understanding of epithelial-mesenchymal transition (EMT) in the setting of tissue fibrosis is largely based on pioneering studies in the kidney. Evidence is emerging that EMT is a key component of chronic kidney disease, contributing to both destruction of the tubular epithelial compartment and accumulation of interstitial fibroblasts. While knowledge regarding EMT was previously based on experimental rodent studies, in recent years the evolving evidence demonstrates a role for EMT in human kidney diseases with chronic fibrosis. PMID- 18508711 TI - Electro-stimulation of cerebellar fastigial nucleus (FNS) improves axonal regeneration. AB - This study focused on the effect of electro-stimulation of fastigial nucleus on the expression of NgR and on axonal regeneration after focal cerebral ischemia reperfusion in rats. Cerebral ischemia and reperfusion was induced by nylon monofilament. Ninety-six male SD rats were randomly divided into sham group and ischemic insult groups at 12 hours, 24 hours, and 1 to 3 weeks after cerebral ischemia-reperfusion. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the changes of NgR mRNA expression. Immunohistochemistry was used to detect the expression of NgR protein and the state of axonal regeneration. Fastigial nucleus stimulation was applied at 2 hours after ischemia for one hour. The results demonstrated that NgR mRNA and protein in the infarcted cortex and hippocampus were significantly increased (p<0.01). The axons were grossly damaged at 24 h after cerebral ischemia-reperfusion when compared to the sham group. Fastigial nucleus stimulation decreased NgR mRNA and protein levels in the infarcted cortex and hippocampus (p<0.01) and improved axonal growth at 24 hours and 2 weeks after ischemia-reperfusion (p<0.05). These results suggest that electrostimulation of fastigial nucleus might provide a new strategy to promote CNS axonal regeneration. PMID- 18508712 TI - Mechanisms of intestinal tight junctional disruption during infection. AB - Tight junctions are dynamic structures that may undergo structural and functional changes in response to both physiological and pathological circumstances. Several microbial pathogens impair intestinal barrier function by exploiting tight junctions. These pathogens have developed a broad and complex range of strategies to subvert host tight junction barrier function. The purpose of this review is to give an overview of the mechanisms whereby select enteric viruses, bacterial pathogens and parasites modulate intestinal tight junctional structure and function and how these effects may contribute to the development of chronic intestinal disorders. PMID- 18508713 TI - The use of genetically engineered model systems for research on human aging. AB - A major goal in the field of aging research is to identify molecular mechanisms of aging at the cellular level, which are anticipated to form the basis for the development of age-associated dysfunctions and diseases in human beings. Recent progress in research into model organisms of aging has allowed determining precise molecular mechanisms and genetic determinants of the aging process, which appear to be conserved in evolution and some of which apply to human aging as well. The consortium of the authors focuses on aging mechanisms at the cellular level, and exploits the potential of genetic analyses in lower eukaryotic model organisms for a better understanding of regulatory pathways implicated in aging processes. We have established a new database (GiSAO), which provides a unique resource for the analysis of genome-wide expression patterns as being regulated by senescence, apoptosis and oxidative stress in our model systems. This has led to the identification of candidate genes, which are being tested for their impact on lifespan regulation in yeast, the fruit fly Drosophila melanogaster and the nematode C. elegans. PMID- 18508714 TI - Advances toward understanding the molecular biology of the Anaplasma-tick interface. AB - The genus Anaplasma includes a diverse group of tick-borne pathogens found exclusively within membrane-bound vacuoles in host cells. While A. marginale, A. centrale and A. ovis, vectored by Dermacentor and Rhipicephalus ticks, are host specific for ruminants, A. phagocytophilum, vectored by Ixodes spp., infects a wide range of hosts. In ticks Anaplasma undergoes a developmental cycle that is coordinated with the tick feeding cycle. Although research at the tick/Anaplasma interface is in its infancy, recent studies have provided evidence that Anaplasma infection and transmission is mediated by a molecular mechanism involving both tick cell and pathogen genes. Application of a growing array of molecular approaches, such as RNA interference, genomics and proteomics, are rapidly expanding our knowledge of the tick/pathogen interface. Targeting key tick cell molecules required for pathogen development in vaccine strategies may compromise the vector capacity of ticks for Anaplasma, thus reducing transmission and infection of vertebrates. Collectively, this information will likely lead to the development of dual target vaccines designed to protect vertebrates against tick infestations and prevent the transmission of pathogens. PMID- 18508715 TI - Molecular characterization and related aspects of the innate immune response in ticks. AB - Compared to insects, little is known about innate immunity in ticks. This chapter addresses the molecular processes that recognize non-self and the cellular and molecular processes mobilized to phagocytose, engulf, inhibit or kill invaders. We discuss the receptors that recognize pathogen associated molecular patterns (PAMPs) and the putative up-regulation of regulatory cascades that lead, ultimately, to cellular or molecular responses. We describe the molecular events that activate the cellular processes and the array of humoral factors that are mobilized against invading organisms, including antimicrobial peptides, proteases and protease inhibitors, lectins, coagulation factors and others. Special attention is directed to the antimicrobial activity of the midgut, the initial site of contact for microbes ingested with the blood. Blood feeding and digestion alone up-regulates an impressive array of proteins, e.g. oxidative stress reducing proteins, lectins, protease inhibitors, proteases, hydrolases, protein/lipid binding agents. Finally, we compare the innate immune responses of ticks with insects and other invertebrates and note deficiencies in our knowledge tick innate immunity. PMID- 18508716 TI - Notch is activated in RANKL-induced osteoclast differentiation and resorption. AB - The process of osteoclast differentiation and resorption is fine-tuned by signal pathways, which need to be further elucidated. The aim of this study was to explore the possible connections between NF-kappaB and Notch in RANKL-induced osteoclast activity. To this end, RANKL was used to stimulate mouse osteoclast precursor cell line RAW264.7. The number of multinucleated TRAP+ osteoclasts was counted and the resorption area was measured. NF-kappaB transcriptional factor activity was determined by EMSA. Quantitative RT-PCR and Western blotting analysis were used to determine Hes1 (one of Notch signaling primary targets) mRNA and protein expressions respectively. Mature osteoclasts and bone resorption areas were detected in the present study. NF-kappaB activity was increased in RANKL-induced osteoclast differentiation and resorption. mRNA and protein expressions of Hes1 in RAW264.7 cells were up-regulated after RANKL stimulation. In conclusion, NF-kappaB signaling mediated RANKL-induced osteoclast differentiation and resorption, during which, Notch signaling was activated. Therefore, Notch could be a novel therapeutic target for bone resorption related diseases. PMID- 18508717 TI - HGF as a renotrophic and anti-fibrotic regulator in chronic renal disease. AB - Hepatocyte growth factor (HGF) and Met/HGF receptor play roles in dynamic growth and morphogenesis during development and regeneration of organs, including the kidney. In the kidney, HGF targets different types of cells, while its biological actions depend on a target cell type. During the earlier stages of chronic renal failure, renal HGF expression increased, but in later stages HGF expression decreased, associated with manifestation of chronic renal failure. When anti-HGF IgG was administered into mice with chronic renal failure, renal dysfunction and fibrosis were accelerated, indicating a role of endogenous HGF to suppress chronic renal failure. For myofibroblasts, a key cell type in tissue fibrosis, HGF exerted biological activities, including (i) inhibition of growth, (ii) suppression of fibrogenic cytokine expression, and (iii) enhancement of matrix metalloproteinases involved in subsequent apoptosis. In models of glomerular and tubulo-interstitial fibrosis, administration of HGF or HGF gene therapy improved renal fibrosis and dysfunction. Since insufficient production of HGF is causative for renal fibrosis, supplementation with HGF represents a new approach to inhibit or improve chronic renal failure. PMID- 18508718 TI - PDE5 modulates oocyte spontaneous maturation via cGMP-cAMP but not cGMP-PKG signaling. AB - Phosphodiesterase type 5 (PDE5), a cGMP specific, cGMP binding phosphodiesterase, specifically hydrolyzes cGMP to 5'-GMP. Here, we examine the distribution of PDE5 in mouse ovary and its effects on spontaneous maturation of mouse oocytes. PDE5 is present in oocytes and cumulus cells of big, antral follicles. Inhibition of activity of PDE5 significantly and reversibly inhibits spontaneous maturation of cumulus-oocyte complexes (COCs). Suppressive effect of PDE5 on spontaneous maturation of COCs is not blocked by the inhibitor of cGMP-dependent protein kinase (PKG). While Sildenafil, an inhibitor of PDE5, has a poor effect on cGMP levels, it significantly increases cAMP levels. These results suggest that the activity of PDE5 plays a role in regulating spontaneous maturation of mouse oocytes and imply that an interaction between cGMP and cAMP signal is involved in this process. PMID- 18508719 TI - Early signals after stretch leading to cardiac hypertrophy. Key role of NHE-1. AB - The enhanced activity of the cardiac Na+/H+ exchanger (NHE-1) after myocardial stretch is considered a key step of the intracellular signaling pathway leading to the slow force response to stretch as well as an early signal for the development of cardiac hypertrophy. We propose that the chain of events triggered by stretch begins with the release of small amounts of Angiotensin II (Ang II)/endothelin (ET) and ends with the increase in intracellular Ca2+ concentration ([Ca2+]i) through the Na+/Ca2+ exchanger in reverse mode (NCX(rev)), which triggers cardiac hypertrophy by activation of widely recognized Ca2+-dependent intracellular signaling pathways. PMID- 18508720 TI - Phytate reduces age-related cardiovascular calcification. AB - The aim of this research was to evaluate the effect of dietary phytate on cardiovascular calcification in rats during aging. Male Wistar rats (10 weeks old) were randomly assigned to four diet groups. The control group was fed with a balanced diet (UAR-A04) containing phytate. The AIN group was fed a purified diet (AIN-76A) with an undetectable level of phytate. The PHY group was fed with a purified diet (AIN-76A) enriched with phytate (phytin, as the calcium magnesium salt). The MOD group was fed with the AIN-76A diet (phytate undetectable) enriched with MgO, inositol and CaHPO4. At 76 weeks of age all rats were sacrificed, and the aortas, hearts, kidneys, livers and femurs were removed for chemical analysis. The most significant differences were found in the aorta calcium content. Phytate-treated rats (the control and PHY groups) had significantly lower levels of calcium in the aorta compared to nonphytate-treated rats (AIN and MOD groups). The present study demonstrated that dietary phytate treatment significantly reduced age-related aorta calcification. PMID- 18508721 TI - FAAP, a novel murine protein, is involved in cell adhesion through regulating vinculin-paxillin association. AB - Focal adhesion associated protein (FAAP), encoded by murine D10Wsu52e gene, is highly homologous to human HSPC117, which interacts with vinculin and talin. HeLa cells transfected with FAAP exhibited normal adhesion incorporation but showed impaired cell spreading, and restrained focal adhesion translocation. Moreover, FAAP facilitated vinculin-paxillin association, decreased interaction of paxillin focal adhesion kinase and inhibited the phosphorylation of extracellular signal regulated kinase. Together, these results suggest that FAAP, by virtue of modulating interaction of adhesion molecules, regulates cell adhesion dynamics. PMID- 18508722 TI - PML NBs (ND10) and Daxx: from nuclear structure to protein function. AB - Proteins that combine PML NBs (ND10) can be divided into two groups: "transient" (that accumulate at PML NBs upon over-expression, interferon-induced up regulation, block of proteosomal degradation, environmental stress or viral infection) and "constitutive" that co-localize with PML in the majority of cultured cells. One of the few "constitutive" components of PML NBs is the death domain-associated protein Daxx. While PML NBs are the most obvious depositories of Daxx, there are multiple alternative localization of this protein in the nucleus and cytoplasm, suggesting differential functionality of Daxx at different cellular compartments and stages of the cell cycle. The purpose of this review is to analyze Daxx spatiotemporal behavior within and outside of PML NBs and to discuss functions attributed to these localizations. We suggest that Daxx can participate in numerous cellular functions as a mediator of protein interactions, thus acting as a fine tuning instrument in highly orchestrated cellular processes; we also envision PML NBs accumulation of Daxx as an "out of action" storage depot. PMID- 18508723 TI - Higher expression of Bax in regulatory T cells increases vascular inflammation. AB - This study is to examine our hypothesis that CD4+CD25(high)Foxp3+ regulatory T cells (Tregs) have an interleukin-2 (IL-2) withdrawal-triggered apoptosis pathway, and modulation of Treg apoptosis pathway affects development of vascular inflammation. We found that pro-apoptotic protein Bax upregulation in Tregs is induced by IL-2 withdrawal. Treg apoptosis induced by IL-2 withdrawal is inhibited by a Bax inhibitor, suggesting that highly expressed Bax is functional. To define the role of upregulated Bax in Treg apoptosis, we established a Tregs specific Bax transgenic mouse model. Enforced expression of Bax in Tregs promotes Treg apoptosis triggered by IL-2 withdrawal and other apoptosis stimuli, suggesting pro-apoptotic role of highly expressed Bax in wild-type Tregs. Finally, higher expression of Bax in Tregs decreases the striking threshold of vascular inflammation due to the failure of suppression of inflammatory cells resulting from Treg apoptosis. These results have demonstrated the proof of principle that the modulation of Tregs apoptosis/survival could be used as a new therapeutic approach for inflammatory cardiovascular diseases. PMID- 18508724 TI - Pancreas islets in metabolic signaling--focus on the beta-cell. AB - The Islets of Langerhans form a nutrient sensing network spread throughout the pancreas. They are tightly connected to the source organ, the intestine, and the target organs--liver, muscle, and fat cells. The expression of a unique set of proteins enables beta cells, the most frequent islet cell type, to detect elevated blood glucose levels and secrete insulin accordingly. Clustered beta cells achieve tighter regulation of glucose-induced insulin secretion by coordination through cell surface proteins. They also adjust their secretory capacity and flow to avoid being damaged. The immediate reaction of the beta cell to nutrients is regulated by translational mechanisms, while longer term adaptations involve changes in transcription. Glucose increases overall protein synthesis in beta-cells but selectively boosts translation of some secretory proteins including insulin. This may be mediated through recognition of RNA motifs in the untranslated regions of those messengers. If essential molecular components of this nutrient sensing system are broken or fail due to repeated stress, beta cells malfunction, which on a larger scale manifest as diseases like diabetes mellitus. PMID- 18508725 TI - Hepatitis E: a curious zoonosis. AB - Hepatitis E virus (HEV) is responsible for large waterborne epidemics of acute hepatitis in endemic regions and for sporadic autochthonous cases in non endemic regions. Although the water vector has been thoroughly documented in endemic regions, very little is known about the modes of contamination occurring in non endemic regions. Unlike the other hepatitis viruses, HEV has an animal reservoir. Several lines of evidence, such as the results of phylogenic analysis and studies on direct contamination via infected food products, have suggested that some cases of animal to human transmission occur. However, all the possible sources of human contamination in non endemic areas have not yet been defined, and this point needs to be investigated. The high genetic variability of HEV, which might be an important factor, involved in zoonotic contamination processes, also needs a surveillance plan. PMID- 18508726 TI - Role of proteasomes in transcription and their regulation by covalent modifications. AB - The 26S proteasome is an abundant multi-subunit complex, which, in addition to lysosomes, represents a major cellular "protein degradation factory". The proteasome complex possesses protease, ATPase/helicase, and RNAse enzymatic activities, which are used by the latter to regulate various physiological processes. Recent findings have revealed an important role of proteasomes in transcriptional regulation. Although proteasomes are well documented to undergo various post-translational modifications, little is known about their functional significance, in particular in the process of gene regulation in response to various forms of stress. Here, we review the data on the role of proteasomes in gene regulation and their post-translational modifications as well as discuss potential mechanisms by which proteasomal activity may be regulated by genotoxic stress. PMID- 18508727 TI - Lefty peptides, derived by MMP2 cleavage, act as a new class of gelatinase A inhibitor. AB - MMPs are zinc-dependent endopeptidases that are involved in proteolysis of extracellular matrix in both physiological and pathological processes including cancer. MMPs are involved at all stages of tumor progression, including tumor growth, angiogenesis, and metastasis. We recently showed that overexpression of Lefty in cancer cells restrains tumor growth. Here, we show that small forms of Lefty are generated by MMP2 (gelatinase A) mediated cleavage. In turn, these forms of Lefty strongly inhibit the autocatalytic, gelatinolytic and caseinolytic activities of MMP2 in vitro. We show that a short synthesized form of Lefty peptide (CASDGALVP) inhibits gelatinolytic and caseinolytic activities of MMP2 in vitro and inhibits tumor growth in vivo. Together, these findings show that lefty peptides are a new class of gelatinase A inhibitors that restrain tumor growth. PMID- 18508728 TI - Frequency-dependence of the slow force response. AB - Stretch induces biphasic inotropic effects in mammalian myocardium. A delayed component (slow force response, SFR) has been demonstrated in various species, however, experimental conditions varied and the underlying mechanisms are controversial. The physiological relevance of the SFR is poorly understood. Experiments were performed in ventricular muscle strips from failing human hearts and non-failing rabbit hearts. Upon stretch, twitch force was assessed at basal conditions (1 Hz, 37 degrees C) and after changing stimulation frequency with and without blockade of the Na+/H+-exchanger-1 (NHE1) or reverse-mode Na+/Ca2+ exchange (NCX). Action potential duration (APD) was assessed using floating electrodes. Low stimulation rates (0.2 Hz) potentiated and higher stimulation rates (2 and 3 Hz) reduced the SFR. The extent of SFR inhibition by NHE1 or NCX inhibition was not affected by stimulation rate. APD decreased at 0.2 Hz but was not altered at higher stimulation rates. The data demonstrate frequency dependence of the SFR with greater positive inotropic effects at lower stimulation rates. Subcellular mechanisms underlying the SFR are not fundamentally affected by stimulation rate. The SFR may have more pronounced physiological effects at lower heart rates. PMID- 18508730 TI - Secreted proteins of Candida albicans. AB - The predicted secretome of the opportunistic fungal pathogen Candida albicans contains more than 200 ORFs of diverse and often unknown function. Majority of the secreted proteins that have been experimentally evaluated to date are hydrolytic enzymes (proteinases, phospholipases and lipases). Acting on the interface between the pathogen and the host, the secreted hydrolases may enable invasion of host tissues, help the pathogen to avoid host defense mechanisms, or allow the microorganism to utilize host cell macromolecules as a source of nutrients. Aspartic proteinases constitute the best-characterized family of the C. albicans secreted proteins. Number of studies addressed also secreted phospholipases. Lipases and N-acetylhexosaminidase have received less attention thus far. Many ORFs that are predicted to encode secreted proteins await characterization. PMID- 18508731 TI - Influence of TBK-1 on tumor angiogenesis and microvascular inflammation. AB - Tank-Binding-Kinase 1 (TBK-1) has been proposed as a putative mediator in tumor angiogenesis. It was the aim of our study to gain insight into TBK-1s role in tumor angiogenesis and tumor-associated microvascular inflammation. TBK-1 overexpressing KB 3-1 cells were generated and their growth characteristics were analyzed. Expression of TBK-1, VEGF, RANTES and Il-8 were quantified using qPCR and western blot analysis. Intravital microscopy using the dorsal skinfold chamber model in nude mice addressed total (TIVD) and functional intratumoral vascular density (FIVD), perfusion index, vessel diameter and leukocyte sticking. Transfection of KB-3 cells resulted in significantly increased TBK-1, RANTES and IL-8 expression without affecting cellular growth. Supernatants from TBK-1 overexpressing clones induced HUVEC proliferation. Intravital microscopy identified an increase in leukocyte sticking paralleled by significantly increased TIVD and FIVD as a result of increased VEGF expression. Therefore, TBK 1 represents a novel mediator of tumor angiogenesis and exerts proinflammatory effects via upregulation of inflammatory cytokines. The TBK-1 pathway might be an important cross-link between angiogenesis and inflammation representing a possible target for anti-tumor therapy. PMID- 18508729 TI - Oxidative stress-induced disruption of epithelial and endothelial tight junctions. AB - Mounting body of evidence indicates that the disruption of epithelial tight junctions and resulting loss of barrier function play a crucial role in the pathogenesis of a variety of gastrointestinal, hepatic, pulmonary, kidney and ocular diseases. Increased production of inflammatory mediators such as cytokines and reactive oxygen species disrupt the epithelial and endothelial barrier function by destabilizing tight junctions. Oxidative stress induced by various reactive oxygen species such as hydrogen peroxide, nitric oxide, peroxynitrite and hypochlorous acid disrupt the epithelial and endothelial tight junctions in various tissues. The mechanism involved in oxidative stress-induced disruption of tight junction includes protein modification such as thiol oxidation, phosphorylation, nitration and carbonylation. The role of signaling molecules such as protein kinases and protein phosphatases in regulation of tight junctions is discussed in this article. Understanding such mechanisms in oxidative stress induced disruption of epithelial and endothelial barrier functions is likely to provide insight into the pathogenesis of various inflammatory diseases, and may form a basis for the design of treatment strategies for different diseases. PMID- 18508732 TI - Hormonal regulation of metamorphosis and reproduction in ticks. AB - The presence of a "status quo" hormone like JH has not been found in ticks. The most advanced understanding of tick endocrinology is associated with female reproduction, where the sequence of the first messages for storage proteins (vitellogenin (Vg) and carrier protein), the Vg receptor, and male peptidic pheromones were recently reported. The current consensus model suggests that ecdysteroids from the epidermis regulated by a putative peptidic ecdysiotrophic hormone from the synganlion initiates the expression of the Vg messages in fat body and midgut. Vg protein, secreted into the hemolymph, requires an ovary Vg receptor to be absorbed by oocytes. Male pheromones transferred into the female genital tract during mating initiate blood feeding to repletion and vitellogenesis. The work so far on tick endocrinology is limited by the paucity of identified hormones and the small number of studies on a few tick models. The role of storage proteins in the evolution of hematophagy is discussed. PMID- 18508733 TI - PsL-EGFmAb inhibits the stimulatory functions of human dendritic cells via DC SIGN. AB - Dendritic cell (DC)-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) is a DC-specific C-type lectin that plays an important role in recognizing and capturing pathogens, DC migration and initiation of T cell responses. Here, we show that anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb), originally prepared for blockade of the adhesive molecule P-selectin, significantly down-regulated DC-SIGN expression as well as expression of mature DC-related molecules including CD83, CD86 and CD80 on human DCs. This PsL-EGFmAb treatment of DCs resulted in impaired allogeneic T cell proliferation and IL-12 production. Furthermore, we show that PsL-EGFmAb-induced down regulation of DC-SIGN may inhibit NF-kappaB expression in DCs, which accounts for the inhibition of DC maturation and stimulatory function. Our present studies indicate that PsL-EGFmAb may be a useful reagent for regulating DC-SIGN expression and DC function. PMID- 18508734 TI - [Personality of a politician]. AB - The present study points to some fundamental connections summarizing the international literature on the personality and intellect of politicians and adapting it to local/home conditions. AIM/OBJECTIVE: The objective is to complete Hungarian knowledge/competence as well as defining new pieces of information and connections on the basis of which further examinations and surveys can be executed. METHOD: Having collected research material for nearly a year, authors have drawn from several publications, books, dissertations, papers and lectures. RESULTS: Theory and investigations of some excellent scientists are reviewed in this essay. The 14 political personality dimensions are analysed and classified according to the factors formed by the authors, which correlated with stylistic factors. CONCLUSIONS: In accordance with the authors, sociologists and psychiatrists have drawn the conclusion that the personality of a politician is determinant in politics as well as in the performance of political leaders. Also, personality correlates with attitude and behaviour. PMID- 18508735 TI - [Clinical aspects of hepatitis C in women of child-bearing age]. AB - It is difficult to define the optimal timing of antiviral treatment in women infected with viral hepatitis C, who have child-bearing potential. Antiviral treatment is strictly contraindicated during pregnancy and the breast-feeding period. Data are conflicting about the question of treatment with modern drugs (peginterferon and ribavirin) before or after pregnancy. The risk of vertical transmission from mother to child is estimated about 5%. The mother's viraemia seems to be the main transmission factor. There is a worse prognosis in nulliparous and postmenopausal women in the natural history of viral hepatitis C. Poor outcome in gestational age, maturity and Apgar score were not associated with hepatitis C virus infection. Combined treatment has frequent gynecological and other side effects. The timing of antiviral therapy in women in child-bearing period is recommended individually. PMID- 18508736 TI - [Legal regulation in the field of tissue engineering]. AB - In the past few years, biotechnology and tissue engineering have developed a great deal. Legislation, however, has not been able to keep pace with the rapid changes in the research and treatment using tissue-engineered products. The authors tried to explore and compare all the regulatory factors concerning cell and tissue engineering in the Hungarian and foreign law. The thorough study elucidated that the Hungarian regulation in this area is fairly deficient when compared with the regulations of other European countries, where quite extensive directives are applied. A currently accepted regulation fills this void efficiently in the European Union providing strong base for tissue engineering related research and therapy for the future. PMID- 18508737 TI - [New generation of abdominal surgery: the Natural Orifice Transluminal Endoscopic Surgery]. AB - Minimal invasive surgical techniques, namely laparoscopic procedures to the abdominal surgery have been introduced more than 20 years ago. In view of clinical results, the majority of these are considered as routinely performed procedure today. Natural Orifice Transluminal Endoscopic Surgery can be considered as a new generation of abdominal surgery. It means a method directed through natural orifices, and abdominal surgery operated by transluminal endoscopic techniques (transgastric, transcolonic, transvaginal) to reach diagnostic and therapeutic goals. Theoretically, this method allows the possibility to decrease invasiveness and postoperative pain, to prevent postoperative hernias and to improve cosmetic results. At present numerous researchers work worldwide to receive scientifically based answers to arising questions (surgical technique, indication, contraindication, complications, monitoring) and concerns in this area. The present paper gives an overview of the national and international literature on experimental results and clinical approaches in the field of this new surgical technique. PMID- 18508738 TI - [Role of heart transplantation in pediatric heart surgery. The first successful pediatric heart transplantation in Hungary]. AB - 7-year-old boy, who underwent aortic valve replacement two years previously, suffered from idiopathic dilated cardiomyopathy. Because of poor condition (NYHA IV), heart transplantation was performed on 18th October 2007. It was the first pediatric heart transplantation in Hungary. It was an uneventful early postoperative period, 6 months after the operation he is doing well, no biopsy proven and tissue Doppler echocardiography (TDI-derived velocities measurement) rejection was detected. The immunosuppression was based on triple-drug therapy (tacrolimus+mycophenolate mofetil+corticosteroid) with use of induction therapy with interleukin-2 receptor blocker (basiliximab). PMID- 18508739 TI - [Iatrogenic injuries]. PMID- 18508740 TI - [Remembering Kalman Buday, professor of pathology (1863-1937)]. PMID- 18508741 TI - [Karoly Borszeky died 75 years ago]. PMID- 18508745 TI - Concealed weapons: erectile claws in African frogs. AB - Vertebrate claws are used in a variety of important behaviours and are typically composed of a keratinous sheath overlying the terminal phalanx of a digit. Keratinous claws, however, are rare in living amphibians; their microstructure and other features indicate that they probably originated independently from those in amniotes. Here we show that certain African frogs have a different type of claw, used in defence, that is unique in design among living vertebrates and lacks a keratinous covering. These frogs have sectorial terminal phalanges on their hind feet that become functional by cutting through the skin. In the resting state, the phalanx is subdermal and attached to a distal bony nodule, a neomorphic skeletal element, via collagen-rich connective tissue. When erected, the claw breaks free from the nodule and pierces the ventral skin. The nodule, suspended by a sheath attached to the terminal phalanx and supported by collagenous connections to the dermis, remains fixed in place. While superficially resembling the shape of claws in other tetrapods, these are the only vertebrate claws known to pierce their way to functionality. PMID- 18508746 TI - Boolean network models of cellular regulation: prospects and limitations. AB - Computer models are valuable tools towards an understanding of the cell's biochemical regulatory machinery. Possible levels of description of such models range from modelling the underlying biochemical details to top-down approaches, using tools from the theory of complex networks. The latter, coarse-grained approach is taken where regulatory circuits are classified in graph-theoretical terms, with the elements of the regulatory networks being reduced to simply nodes and links, in order to obtain architectural information about the network. Further, considering dynamics on networks at such an abstract level seems rather unlikely to match dynamical regulatory activity of biological cells. Therefore, it came as a surprise when recently examples of discrete dynamical network models based on very simplistic dynamical elements emerged which in fact do match sequences of regulatory patterns of their biological counterparts. Here I will review such discrete dynamical network models, or Boolean networks, of biological regulatory networks. Further, we will take a look at such models extended with stochastic noise, which allow studying the role of network topology in providing robustness against noise. In the end, we will discuss the interesting question of why at all such simple models can describe aspects of biology despite their simplicity. Finally, prospects of Boolean models in exploratory dynamical models for biological circuits and their mutants will be discussed. PMID- 18508747 TI - A common tendency for phylogenetic overdispersion in mammalian assemblages. AB - Competition has long been proposed as an important force in structuring mammalian communities. Although early work recognized that competition has a phylogenetic dimension, only with recent increases in the availability of phylogenies have true phylogenetic investigations of mammalian community structure become possible. We test whether the phylogenetic structure of 142 assemblages from three mammalian clades (New World monkeys, North American ground squirrels and Australasian possums) shows the imprint of competition. The full set of assemblages display a highly significant tendency for members to be more distantly related than expected by chance (phylogenetic overdispersion). The overdispersion is also significant within two of the clades (monkeys and squirrels) separately. This is the first demonstration of widespread overdispersion in mammal assemblages and implies an important role for either competition between close relatives where traits are conserved, habitat filtering where distant relatives share convergent traits, or both. PMID- 18508748 TI - Making it on their own: sperm-dependent hybrid fishes (Cobitis) switch the sexual hosts and expand beyond the ranges of their original sperm donors. AB - Interspecific hybridization may result in asexual hybrid lineages that reproduce via parthenogenesis. Contrary to true parthenogens, sperm-dependent asexuals (gynogens and hybridogens) are restricted to the range of bisexual species, generally the parental taxa, by their need for a sperm donor. It has been documented that asexual lineages may rarely use sperm from a non-parental species or even switch a host. The available literature reports do not allow distinguishing, between whether such host switches arise by the expansion of asexuals out of their parental's range (and into that of another's) or by the local extinction of a parental population followed by a host switch. The present study combines new and previously collected data on the distribution and history of gynogenetic spined loaches (Cobitis) of hybrid origin. We identified at least three clonal lineages that have independently switched their sperm dependency to different non-parental Cobitis species, and in cases incorporated their genomes. Our current knowledge of European Cobitis species and their hybrids suggests that this pattern most probably results from the expansion of gynogenetic lineages into new areas. Such expansion was independent of the original parental species. This suggests that sperm dependence is not as restrictive to geographical expansion when compared with true parthenogenesis as previously thought. PMID- 18508749 TI - The ubiquitous mountain hare mitochondria: multiple introgressive hybridization in hares, genus Lepus. AB - Climatic oscillations during the glaciations forced dramatic changes in species distributions, such that some presently temperate regions were alternately occupied by temperate and arctic species. These species could have met and hybridized during climatic transitions. This phenomenon happened for three hare species present in Iberia (Lepus granatensis, Lepus europaeus and Lepus castroviejoi), which display high frequencies of mitochondrial DNA (mtDNA) from Lepus timidus, an arctic/boreal species presently extinct in Iberia. Here, we extend our previous geographical survey to determine whether the distribution of this mtDNA lineage extends beyond the northern half of the Iberian Peninsula, where it is found at high frequencies. We also review the taxonomy, distribution and molecular phylogeny of the genus Lepus. The phylogenetic inference reveals the presence of L. timidus-like mtDNA in several other hare species in Asia and North America, suggesting that the mitochondrial introgression observed in Iberia might be generalized. Comparison with the available nuclear gene phylogenies suggests that introgression could have happened repeatedly, possibly during different climatic transitions. We discuss demographic and adaptive scenarios that could account for the repetition in time and space of this spectacular phenomenon and suggest ways to improve our understanding of its determinants and consequences. Such high levels of introgressive hybridization should discourage attempts to revise hare taxonomy based solely on mtDNA. PMID- 18508750 TI - Fission and fusion of Darwin's finches populations. AB - This study addresses the causes and evolutionary consequences of introgressive hybridization in the sympatric species of Darwin's ground finches (Geospiza) on the small island of Daphne Major in the Galapagos archipelago. Hybridization occurs rarely (less than 2% of breeding pairs) but persistently across years, usually as a result of imprinting on the song of another species. Hybrids survive well under some ecological conditions, but not others. Hybrids mate according to song type. The resulting introgression increases phenotypic and genetic variation in the backcrossed populations. Effects of introgression on beak shape are determined by the underlying developmental genetic pathways. Introgressive hybridization has been widespread throughout the archipelago in the recent past, and may have been a persistent feature throughout the early history of the radiation, episodically affecting both the speed and direction of evolution. We discuss how fission through selection and fusion through introgression in contemporary Darwin's finch populations may be a reflection of processes occurring in other young radiations. We propose that introgression has the largest effect on the evolution of interbreeding species after they have diverged in morphology, but before the point is reached when genetic incompatibilities incur a severe fitness cost. PMID- 18508751 TI - Male territorial vocalizations and responses are decoupled in an avian hybrid zone. AB - A core area of speciation research concerns the coevolution of species-specific signals and the selective sensitivity to such signals. Signals and responses to them should be tuned to each other, to be effective in intraspecific communication. Hybrid zones are ideal to study the presence of such 'behavioural coupling' and the mechanisms governing it, and this has rarely been done. Our study examines acoustic signals of males and their response to them in the context of territorial interactions in a natural hybrid zone between two dove species, Streptopelia vinacea and Streptopelia capicola. Male signals are important in hybrid zone dynamics as they are essential for territory establishment, which is crucial for successful reproduction. We tested whether the response of individual male hybrids is linked to how similar their own signal is to the playback signal. We did not find evidence for behavioural coupling. The combined evidence from the low level of response to hybrid and heterospecific signals outside the hybrid zone and a lack of coupling within the hybrid zone suggests that perceptual learning may explain our results. Learning to respond to locally abundant signals may be the best individual strategy and is likely to contribute to the maintenance of a hybrid zone. PMID- 18508752 TI - Female mating preference functions predict sexual selection against hybrids between sibling species of cichlid fish. AB - The evolutionary outcome of interspecific hybridization, i.e. collapse of species into a hybrid swarm, persistence or even divergence with reinforcement, depends on the balance between gene flow and selection against hybrids. If female mating preferences are open-ended but sign-inversed between species, they can theoretically be a source of such selection. Cichlid fish in African lakes have sustained high rates of speciation despite evidence for widespread hybridization, and sexual selection by female choice has been proposed as important in the origin and maintenance of species boundaries. However, it had never been tested whether hybridizing species have open-ended preference rules. Here we report the first experimental test using Pundamilia pundamilia, Pundamilia nyererei and their hybrids in three-way choice experiments. Hybrid males are phenotypically intermediate. Wild-caught females of both species have strong preferences for conspecific over heterospecific males. Their responses to F1 hybrid males are intermediate, but more similar to responses to conspecifics in one species and more similar to responses to heterospecifics in the other. We suggest that their mate choice mechanism may predispose haplochromine cichlids to maintain and perhaps undergo phenotypic diversification despite hybridization, and that species differences in female preference functions may predict the potential for adaptive trait transfer between hybridizing species. PMID- 18508753 TI - Spatial, environmental and anthropogenic effects on the taxon composition of hybridizing Daphnia. AB - The competitive ability of hybrids, compared with their parental taxa, can cover a wide fitness range from poor to superior. For example communities of the Daphnia galeata-hyalina-cucullata species complex often show hybrid dominance. We tested whether taxa composition of 43 European lakes inhabited by this species complex can be explained by habitat characteristics (e.g. size descriptors, trophy level) or geography. We found that D. galeata occurs more frequently south of the Alps, whereas D. hyalina and D. cucullata are found more in the north. Lakes with D. galeata dominance had higher temperatures whereas D. hyalina dominance could be attributed to low phosphorus loads. The dominance of F1 hybrids, however, was not explainable with current environmental variables. In a subset of 28 lakes, we studied the impact of eutrophication history on F-hybrid success. Lakes with the highest trophic state in the past tended to be dominated by F1-hybrids. Our data demonstrate that human-mediated habitat disturbance (eutrophication) has facilitated hybrid success and altered the Daphnia taxon composition across lakes. At the same time, specific habitat conditions might provide a refuge from hybridization for native genotypes. PMID- 18508754 TI - Bayesian inference of species hybrids using multilocus dominant genetic markers. AB - Neutral genetic markers are useful for identifying species hybrids in natural populations, especially when used in conjunction with statistical methods like the one implemented in the software NEWHYBRIDS. Here, a short description of the extension of NEWHYBRIDS to dominant markers is given. Subsequently, an extensive series of simulations of amplified fragment length polymorphism (AFLP) data is performed to evaluate the prospects for hybrid identification with (possibly non diagnostic) dominant markers. Distinguishing between F1's and F2's is shown to be difficult, possibly requiring upwards of 100 AFLP markers to be done accurately. Discriminating between pure-bred and non-pure (hybrid) individuals, however, is shown to be much easier, requiring perhaps as few as 10 dominant markers, even from relatively weakly diverged species. PMID- 18508755 TI - Hybridization of glaucous gull (Larus hyperboreus) and herring gull (Larus argentatus) in Iceland: mitochondrial and microsatellite data. AB - Large white-headed gulls provide an interesting group of birds for studies of hybridization. The group is composed of 20 species of recent origin, often with weak reproductive barriers. Here we report the results from a study on the glaucous gull Larus hyperboreus, an Arctic species which has been breeding in Iceland for centuries, and the herring gull Larus argentatus which has a wide distribution in Europe but colonized Iceland in 1920s. Previous studies, based on morphological variation indicated hybridization between the two species in Iceland, have been questioned as it may just reflect variation within the species. Here we evaluate whether hybridization has occurred between the two species in Iceland by studying variation in microsatellites and mtDNA. The analysis is based on feathers taken from wings sampled in Iceland over a period of 40 years. The results are compared with samples obtained from East Greenland and published sequences of samples obtained throughout Europe. The genetic analysis reveals a distinctive grouping of the two species, although they present a shallow genealogy and an extensive sharing of the genetic variants between the two species. Several individuals show admixture for molecular markers, which may result from an incomplete lineage sorting although geographical patterns of both mtDNA haplotypes and microsatellites strongly indicate a recent hybridization in Iceland. PMID- 18508756 TI - The origin and evolution of a unisexual hybrid: Poecilia formosa. AB - Clonal reproduction in vertebrates can always be traced back to hybridization events as all known unisexual vertebrates are hybrids between recognized species or genetically defined races. Interestingly, clonal vertebrates often also rely on interspecific matings for their reproduction because gynogenesis (sperm dependent parthenogenesis) and hybridogenesis are common modes of propagation. While in most cases these hybridization events leave no hereditary traces in the offspring, occasionally the genome exclusion mechanism fails and either small parts of male genetic material remain inside the oocyte in the form of microchromosomes, or fusion of the sperm nucleus with the oocyte nucleus leads to polyploid individuals. In this review, we highlight the important role of hybridization for the origin and evolution of a unisexual hybrid: the Amazon molly, Poecilia formosa. PMID- 18508757 TI - Review. Lifelong commitment to the wrong partner: hybridization in ants. AB - The extraordinary lifelong partner commitment in social insects is expected to increase choosiness in both sexes and therefore to be associated with particularly low hybridization frequencies. Yet, more and more studies reveal that in many ant taxa hybrids are surprisingly common, with up to half of all female sexuals receiving sperm from allospecific males in extreme cases. In a few ant species, hybridization has led to the evolution of reproductively isolated new lineages with a bizarre system of genetic caste differentiation: colonies produce hybrid workers and pure-lineage female sexuals. This requires that colonies either contain multiple queens or that queens mate multiple times. In most other cases, hybridization appears to be an evolutionary dead end and fertile hybrid queens are rarely found. In such cases, haplodiploid sex determination appears to decrease the costs of mating with an allospecific male. As long as hybrid workers are viable, a cross-mated queen can partially rescue its fitness by producing males from unfertilized eggs. Mating with an allospecific partner may thus be an option for queens when conspecific mates are not available. The morphological similarity of most ant males, perhaps resulting from the lack of sexual conflict, may similarly contribute to the commoness of hybridization. PMID- 18508758 TI - Daphnia hybridization along ecological gradients in pelagic environments: the potential for the presence of hybrid zones in plankton. AB - The relative homogeneity of pelagic environments has been regarded as the reason for the absence of hybrid zones for hybridizing planktonic Daphnia (Crustacea: Cladocera); occasional dominance of interspecific hybrids over parental species was explained by their temporal superiority in fluctuating environments. However, water bodies with spatially varying environmental conditions might facilitate the formation of hybrid zones in plankton. We studied the distribution of species and hybrids of the Daphnia longispina complex in 11 canyon-shaped reservoirs, localities characterized by horizontal environmental gradients (particularly of food supply and size-selective predation); we also analysed patterns of carapace size and fecundity among coexisting taxa. Spatial distribution of taxa agreed with their ecological characteristics; those showing different affinities along longitudinal reservoir profiles differed in size according to the presumed fish predation gradient. Only hybrids of Daphnia galeata with Daphnia cucullata and D. longispina (=hyalina) were recorded. The latter two species preferred opposite ends of gradients, such spatial segregation probably explaining the absence of their hybrids. Distributional patterns were relatively stable in two consecutive summers, apart from a substantial decline of D. galeata X cucullata in the second year. The observed pattern of a hybrid-dominated zone in intermediate conditions suggests that local Daphnia hybrid zones may indeed form within reservoirs. PMID- 18508759 TI - Mitogen-activated protein kinase phosphatase-1 represses c-Jun NH2-terminal kinase-mediated apoptosis via NF-kappaB regulation. AB - The mechanism regulating radiation-induced anti-apoptotic response, a limiting factor in improving cell radiosensitivity, remains elusive. Mitogen-activated protein kinase (MAPK) phosphatase (MKP)-1 is the major member of MKPs that dephosphorylates and inactivates MAPK. Here we provide the evidence that MKP-1 was negatively bridging between NF-kappaB-mediated prosurvival pathway and c-Jun N-terminal kinase (JNK)-mediated proapoptotic response. MKP-1 was induced by gamma-radiation and repressed radiation-induced pro-apoptotic status. NF-kappaB RelA/p50 heterodimer was recruited to MKP-1 gene promoter to induce MKP-1 transcription. Deletion of the NF-kappaB-binding site or inactivation of NF kappaB by its small interfering RNA significantly decreased the radiation-induced MKP-1 promoter activity. In addition, MKP-1-deficient mouse embryonic fibroblasts exhibited a prolonged activation of JNK but not p38 or extracellular signal regulated kinase subfamilies of MAPKs. The prolonged activation of JNK was not induced by treatment with tumor necrosis factor alpha or interleukin-6, and inactivation of JNK but not p38 or ERK abolished radiation-induced proapoptotic status, indicating that JNK is specifically inhibited by radiation-induced MKP-1. Three MKP-1 wild type human tumor cell lines treated with MKP-1 small interfering RNA showed an increased proapoptotic response that can be rescued by overexpression of wild type mouse MKP-1. Together, these results suggest that MKP 1 is a NF-kappaB-mediated prosurvival effector in attenuating JNK-mediated pro apoptotic response; NF-kappaB/MKP-1-mediated negative JNK regulation represents a potential therapeutic target for adjusting cell radiosensitivity. PMID- 18508760 TI - The molecular basis of functional bacterial amyloid polymerization and nucleation. AB - Amyloid fibers are filamentous proteinaceous structures commonly associated with mammalian neurodegenerative diseases. Nucleation is the rate-limiting step of amyloid propagation, and its nature remains poorly understood. Escherichia coli assembles functional amyloid fibers called curli on the cell surface using an evolved biogenesis machine. In vivo, amyloidogenesis of the major curli subunit protein, CsgA, is dependent on the minor curli subunit protein, CsgB. Here, we directly demonstrated that CsgB(+) cells efficiently nucleated purified soluble CsgA into amyloid fibers on the cell surface. CsgA contains five imperfect repeating units that fulfill specific roles in directing amyloid formation. Deletion analysis revealed that the N- and C-terminal most repeating units were required for in vivo amyloid formation. We found that CsgA nucleation specificity is encoded by the N- and C-terminal most repeating units using a blend of genetic, biochemical, and electron microscopic analyses. In addition, we found that the C-terminal most repeat was most aggregation-prone and dramatically contributed to CsgA polymerization in vitro. This work defines the elegant molecular signatures of bacterial amyloid nucleation and polymerization, thereby revealing how nature directs amyloid formation to occur at the correct time and location. PMID- 18508761 TI - Multiple intrinsically disordered sequences alter DNA binding by the homeodomain of the Drosophila hox protein ultrabithorax. AB - During animal development, distinct tissues, organs, and appendages are specified through differential gene transcription by Hox transcription factors. However, the conserved Hox homeodomains bind DNA with high affinity yet low specificity. We have therefore explored the structure of the Drosophila melanogaster Hox protein Ultrabithorax and the impact of its nonhomeodomain regions on DNA binding properties. Computational and experimental approaches identified several conserved, intrinsically disordered regions outside the homeodomain of Ultrabithorax that impact DNA binding by the homeodomain. Full-length Ultrabithorax bound to target DNA 2.5-fold weaker than its isolated homeodomain. Using N-terminal and C-terminal deletion mutants, we demonstrate that the YPWM region and the disordered microexons (termed the I1 region) inhibit DNA binding approximately 2-fold, whereas the disordered I2 region inhibits homeodomain-DNA interaction a further approximately 40-fold. Binding is restored almost to homeodomain affinity by the mostly disordered N-terminal 174 amino acids (R region) in a length-dependent manner. Both the I2 and R regions contain portions of the activation domain, functionally linking DNA binding and transcription regulation. Given that (i) the I1 region and a portion of the R region alter homeodomain-DNA binding as a function of pH and (ii) an internal deletion within I1 increases Ultrabithorax-DNA affinity, I1 must directly impact homeodomain-DNA interaction energetics. However, I2 appears to indirectly affect DNA binding in a manner countered by the N terminus. The amino acid sequences of I2 and much of the I1 and R regions vary significantly among Ultrabithorax orthologues, potentially diversifying Hox-DNA interactions. PMID- 18508762 TI - Oncogenic B-RafV600E inhibits apoptosis and promotes ERK-dependent inactivation of Bad and Bim. AB - Recent studies have revealed that B-Raf mutations are very common in malignant melanoma and are required for tumor growth and maintenance. The majority of melanoma-associated B-Raf mutations involve a single point mutation, V600E, which results in greatly elevated B-Raf kinase activity and constitutive activation of MAPK/ERK downstream. Here we show that B-Raf(V600E) increases resistance to apoptosis induced by chemotherapeutic drugs and promotes ERK-dependent phosphorylation of the BH3-only proteins Bim and Bad that are involved in setting thresholds for apoptosis. ERK-dependent phosphorylation of Bim resulted in degradation of this BH3-only protein, whereas phosphorylation of Bad has previously been shown to result in its sequestration by 14-3-3 proteins. Consistent with this, inhibition of ERK activity in a panel of melanoma cell lines resulted in stabilization of Bim and dephosphorylation of Bad. Furthermore, apoptosis induced through overexpression of Bad or Bim was efficiently blocked by coexpression of mutant B-Raf(V600E). However, small interfering RNA-mediated silencing of Bim and Bad expression conferred only modest protection against cytotoxic drugs, whereas oncogenic B-Raf strongly protected against the same stimuli. These observations suggest that B-Raf-initiated inactivation of Bad and Bim only partly contributes to the anti-apoptotic activities of this oncogene and that other points within the cell death machinery are also targeted by deregulated ERK signaling. PMID- 18508763 TI - The catalytic intermediate stabilized by a "down" active site loop for diaminopimelate decarboxylase from Helicobacter pylori. Enzymatic characterization with crystal structure analysis. AB - The meso-diaminopimelate decarboxylase (DAPDC, EC 4.1.1.20) catalyzes the final step of L-lysine biosynthesis in bacteria and is regarded as a target for the discovery of antibiotics. Here we report the 2.3A crystal structure of DAPDC from Helicobacter pylori (HpDAPDC). The structure, in which the product L-lysine forms a Schiff base with the cofactor pyridoxal 5'-phosphate, provides structural insight into the substrate specificity and catalytic mechanism of the enzyme, and implies that the carboxyl to be cleaved locates at the si face of the cofactor. To our knowledge, this might be the first reported external aldimine of DAPDC. Moreover, the active site loop of HpDAPDC is in a "down" conformation and shields the ligand from solvent. Mutations of Ile(148) from the loop greatly impaired the catalytic efficiency. Combining the structural analysis of the I148L mutant, we hypothesize that HpDAPDC adopts an induced-fit catalytic mechanism in which this loop cycles through "down" and "up" conformations to stabilize intermediates and release product, respectively. Our work is expected to provide clues for designing specific inhibitors of DAPDC. PMID- 18508764 TI - The structure of alpha-parvin CH2-paxillin LD1 complex reveals a novel modular recognition for focal adhesion assembly. AB - Alpha-parvin is an essential component of focal adhesions (FAs), which are large multiprotein complexes that link the plasma membrane and actin cytoskeleton. Alpha-parvin contains two calponin homology (CH) domains and its C-terminal CH2 domain binds multiple targets including paxillin LD motifs for regulating the FA network and signaling. Here we describe the solution structure of alpha-parvin CH2 bound to paxillin LD1. We show that although CH2 contains the canonical CH fold, a previously defined N-terminal linker forms an alpha-helix that packs unexpectedly with the C-terminal helix of CH2, resulting in a novel variant of the CH domain. Importantly, such packing generates a hydrophobic surface that recognizes the Leu-rich face of paxillin-LD1, and the binding pattern differs drastically from the classical paxillin-LD binding to four-helix bundle proteins such as focal adhesion kinase. These results define a novel modular recognition mode and reveal how alpha-parvin associates with paxillin to mediate the FA assembly and signaling. PMID- 18508765 TI - Characterization of a novel cardiac isoform of the cell cycle-related kinase that is regulated during heart failure. AB - Myocardial infarction (MI) is often followed by heart failure (HF), but the mechanisms precipitating the transition to HF remain largely unknown. A genomic profile was performed in a monkey model of MI, from the myocardium adjacent to chronic (2-month) MI followed by 3 weeks of pacing to develop HF. The transcript of the gene encoding the cell cycle-related kinase (CCRK) was down-regulated by 50% in HF heart compared with control (p<0.05), which was confirmed by quantitative PCR. The CCRK sequence cloned from a heart library showed a conservation of the N-terminal kinase domain when compared with the "generic" isoform cloned previously but a different C-terminal half due to alternative splicing with frameshift. The homology of the cardiac sequence was 100% between mice and humans. Expression of the corresponding protein, measured upon generation of a monoclonal antibody, was limited to heart, liver, and kidney. Upon overexpression in cardiac myocytes, both isoforms promote cell growth and reduce apoptosis by chelerythrine (p<0.05 versus control). Using a yeast two hybrid screening, we found an interaction of the generic but not the cardiac CCRK with cyclin H and casein kinase 2. In addition, only the generic CCRK phosphorylates the cyclin-dependent kinase 2, which was accompanied by a doubling of myocytes in the S and G(2) phases of the cell cycle (p < 0.05 versus control). Therefore, the heart expresses a splice variant of CCRK, which promotes cardiac cell growth and survival; differs from the generic isoform in terms of protein protein interactions, substrate specificity and regulation of the cell cycle; and is down-regulated significantly in HF. PMID- 18508766 TI - Elongin B/C recruitment regulates substrate binding by CIS. AB - SOCS proteins play a major role in the regulation of cytokine signaling. They are recruited to activated receptors and can suppress signaling by different mechanisms including targeting of the receptor complex for proteasomal degradation. The activity of SOCS proteins is regulated at different levels including transcriptional control and posttranslational modification. We describe here a novel regulatory mechanism for CIS, one of the members of this protein family. A CIS mutant deficient in recruitment of the Elongin B/C complex completely failed to suppress STAT5 activation. This deficiency was not caused by altered turnover of CIS but by loss of cytokine receptor interaction. Intriguingly, no such effect was seen for binding to MyD88. The interaction between CIS and the Elongin B/C complex, which depends on the levels of uncomplexed Elongin B/C, was easily disrupted. This regulatory mechanism may be unique for CIS, as similar mutations in SOCS1, -2, -3, -6, and -7 had no functional impact. Our findings indicate that the SOCS box not only plays a role in the formation of E3 ligase complexes but, at least for CIS, can also regulate the binding modus of SOCS box-containing proteins. PMID- 18508767 TI - Structural and functional characterization of transmembrane segment IX of the NHE1 isoform of the Na+/H+ exchanger. AB - The Na(+)/H(+) exchanger isoform 1 (NHE1) is an integral membrane protein that regulates intracellular pH by removing one intracellular H(+) in exchange for one extracellular Na(+). It has a large N-terminal membrane domain of 12 transmembrane segments and an intracellular C-terminal regulatory domain. We characterized the cysteine accessibility of amino acids of the putative transmembrane segment IX (residues 339-363). Each residue was mutated to cysteine in a functional cysteineless NHE1 protein. Of 25 amino acids mutated, 5 were inactive or nearly so after mutation to cysteine. Several of these showed aberrant targeting to the plasma membrane and reduced expression of the intact protein, whereas others were expressed and targeted correctly but had defective NHE1 function. Of the active mutants, Glu(346) and Ser(351) were inhibited >70% by positively charged [2-(trimethylammonium)-ethyl]methanethiosulfonate but not by anionic [2-sulfonatoethyl]methanethiosulfonate, suggesting that they are pore lining and make up part of the cation conduction pathway. Both mutants also had decreased affinity for Na(+) and decreased activation by intracellular protons. The structure of a peptide representing amino acids 338-365 was determined by using high resolution NMR in dodecylphosphocholine micelles. The structure contained two helical regions (amino acids Met(340)-Ser(344) and Ile(353) Ser(359)) kinked with a large bend angle around a pivot point at amino acid Ser(351). The results suggest that transmembrane IX is critical with pore-lining residues and a kink at the functionally important residue Ser(351). PMID- 18508768 TI - Critical role of PDE4D in beta2-adrenoceptor-dependent cAMP signaling in mouse embryonic fibroblasts. AB - One of the defining properties of beta2-adrenergic receptor (beta(2)AR) signaling is the transient and rapidly reversed accumulation of cAMP. Here we have investigated the contribution of different PDE4 proteins to the generation of this transient response. To this aim, mouse embryonic fibroblasts deficient in PDE4A, PDE4B, or PDE4D were generated, and the regulation of PDE activity, the accumulation of cAMP, and CREB phosphorylation in response to isoproterenol were monitored. Ablation of PDE4D, but not PDE4A or PDE4B, had a major effect on the beta-agonist-induced PDE activation, with only a minimal increase in PDE activity being retained in PDE4D knock-out (KO) cells. Accumulation of cAMP was markedly enhanced, and the kinetics of cAMP accumulation were altered in their properties in PDE4DKO but not PDE4BKO cells. Modest effects were observed in PDE4AKO mouse embryonic fibroblasts. The return to basal levels of both cAMP accumulation and CREB phosphorylation was greatly delayed in the PDE4DKO cells, suggesting that PDE4D is critical for dissipation of the beta2AR stimulus. This effect of PDE4D ablation was in large part due to inactivation of a negative feedback mechanism consisting of the PKA-mediated activation of PDE4D in response to elevated cAMP levels, as indicated by experiments using the cAMP-dependent protein kinase inhibitors H89 and PKI. Finally, PDE4D ablation affected the kinetics of beta2AR desensitization as well as the interaction of the receptor with Galphai. These findings demonstrate that PDE4D plays a major role in shaping the beta2AR signal. PMID- 18508769 TI - Analysis of the membrane topology of transmembrane segments in the C-terminal hydrophobic domain of the yeast vacuolar ATPase subunit a (Vph1p) by chemical modification. AB - The integral V(0) domain of the vacuolar (H(+))-ATPases (V-ATPases) provides the pathway by which protons are transported across the membrane. Subunit a is a 100 kDa integral subunit of V(0) that plays an essential role in proton translocation. To better define the membrane topology of subunit a, unique cysteine residues were introduced into a Cys-less form of the yeast subunit a (Vph1p) and the accessibility of these cysteine residues to modification by the membrane permeant reagent N-ethylmaleimide (NEM) and the membrane impermeant reagent polyethyleneglycol maleimide (PEG-mal) in the presence and absence of the protein denaturant SDS was assessed. Thirty Vph1p mutants containing unique cysteine residues were constructed and analyzed. Cysteines introduced between residues 670 and 710 and between 807 and 840 were modified by PEG-mal in the absence of SDS, indicating a cytoplasmic orientation. Cysteines introduced between residues 602 and 620 and between residues 744 and 761 were modified by NEM but not PEG-mal in the absence of SDS, suggesting a lumenal orientation. Finally, cysteines introduced at residues 638, 645, 648, 723, 726, 734, and at nine positions between residue 766 and 804 were modified by NEM and PEG-mal only in the presence of SDS, consistent with their presence within the membrane or at a protein-protein interface. The results support an eight transmembrane helix (TM) model of subunit a in which the C terminus is located on the cytoplasmic side of the membrane and provide information on the location of hydrophilic loops separating TM6, 7, and 8. PMID- 18508770 TI - Mechanism-based labeling defines the free energy change for formation of the covalent glycosyl-enzyme intermediate in a xyloglucan endo-transglycosylase. AB - Xyloglucan endo-transglycosylases (XETs) are key enzymes involved in the restructuring of plant cell walls during morphogenesis. As members of glycoside hydrolase family 16 (GH16), XETs are predicted to employ the canonical retaining mechanism of glycosyl transfer involving a covalent glycosyl-enzyme intermediate. Here, we report the accumulation and direct observation of such intermediates of PttXET16-34 from hybrid aspen by electrospray mass spectrometry in combination with synthetic "blocked" substrates, which function as glycosyl donors but are incapable of acting as glycosyl acceptors. Thus, GalGXXXGGG and GalGXXXGXXXG react with the wild-type enzyme to yield relatively stable, kinetically competent, covalent GalG-enzyme and GalGXXXG-enzyme complexes, respectively (Gal=Galbeta(1-->4), G=Glcbeta(1-->4), and X=Xylalpha(1-->6)Glcbeta(1-->4)). Quantitation of ratios of protein and saccharide species at pseudo-equilibrium allowed us to estimate the free energy change (DeltaG(0)) for the formation of the covalent GalGXXXG-enzyme as 6.3-8.5 kJ/mol (1.5-2.0 kcal/mol). The data indicate that the free energy of the beta(1-->4) glucosidic bond in xyloglucans is preserved in the glycosyl-enzyme intermediate and harnessed for religation of the polysaccharide in vivo. PMID- 18508771 TI - DOA1/UFD3 plays a role in sorting ubiquitinated membrane proteins into multivesicular bodies. AB - Ubiquitin (Ub) is a sorting signal that targets integral membrane proteins to the interior of the vacuole/lysosome by directing them into lumenal vesicles of multivesicular bodies (MVBs). The Vps27-Hse1 complex, which is homologous to the Hrs-STAM complex in mammalian cells, serves as a Ub-sorting receptor at the surface of early endosomes. We have found that Hse1 interacts with Doa1/Ufd3. Doa1 is known to interact with Cdc48/p97 and Ub and is required for maintaining Ub levels. We find that the Hse1 Src homology 3 domain binds directly to the central PFU domain of Doa1. Mutations in Doa1 that block Hse1 binding but not Ub binding do not alter Ub levels but do result in the missorting of the MVB cargo GFP-Cps1. Loss of Doa1 also causes a synthetic growth defect when combined with loss of Vps27. Unlike the loss of Doa1 alone, the doa1Delta vps27Delta double mutant phenotype is not suppressed by Ub overexpression, demonstrating that the effect is not due to indirect consequence of lowered Ub levels. Loss of Doa1 results in a defect in the accumulation of GFP-Ub within yeast vacuoles, implying that there is a reduction in the flux of ubiquitinated membrane proteins through the MVB pathway. This defect was also reflected by an inability to properly sort Vph1-GFP-Ub, a modified subunit of the multiprotein vacuolar ATPase complex, which carries an in-frame fusion of Ub as an MVB sorting signal. These results reveal novel roles for Doa1 in helping to process ubiquitinated membrane proteins for sorting into MVBs. PMID- 18508772 TI - Conserved aspartic acid residues lining the extracellular loop 1 of sodium coupled bile acid transporter ASBT Interact with Na+ and 7alpha-OH moieties on the ligand cholestane skeleton. AB - Functional contributions of residues Val-99-Ser-126 lining extracellular loop (EL) 1 of the apical sodium-dependent bile acid transporter were determined via cysteine-scanning mutagenesis, thiol modification, and in silico interpretation. Despite membrane expression for all but three constructs (S112C, Y117C, S126C), most EL1 mutants (64%) were inactivated by cysteine mutation, suggesting a functional role during sodium/bile acid co-transport. A negative charge at conserved residues Asp-120 and Asp-122 is required for transport function, whereas neutralization of charge at Asp-124 yields a functionally active transporter. D124A exerts low affinity for common bile acids except deoxycholic acid, which uniquely lacks a 7alpha-hydroxyl (OH) group. Overall, we conclude that (i) Asp-122 functions as a Na(+) sensor, binding one of two co-transported Na(+) ions, (ii) Asp-124 interacts with 7alpha-OH groups of bile acids, and (iii) apolar EL1 residues map to hydrophobic ligand pharmacophore features. Based on these data, we propose a comprehensive mechanistic model involving dynamic salt bridge pairs and hydrogen bonding involving multiple residues to describe sodium dependent bile acid transporter-mediated bile acid and cation translocation. PMID- 18508773 TI - Phosphorylation determines the calmodulin-mediated Ca2+ response and water permeability of AQP0. AB - In Xenopus oocytes, the water permeability of AQP0 (P(f)) increases with removal of external calcium, an effect that is mediated by cytoplasmic calmodulin (CaM) bound to the C terminus of AQP0. To investigate the effects of serine phosphorylation on CaM-mediated Ca(2+) regulation of P(f), we tested the effects of kinase activation, CaM inhibition, and a series of mutations in the C terminus CaM binding site. Calcium regulation of AQP0 P(f) manifests four distinct phenotypes: Group 1, with high P(f) upon removal of external Ca(2+) (wild-type, S229N, R233A, S235A, S235K, K238A, and R241E); Group 2, with high P(f) in elevated (5 mm) external Ca(2+) (S235D and R241A); Group 3, with high P(f) and no Ca(2+) regulation (S229D, S231N, S231D, S235N, and S235N/I236S); and Group 4, with low P(f) and no Ca(2+) regulation (protein kinase A and protein kinase C activators, S229D/S235D and S235N/I236S). Within each group, we tested whether CaM binding mediates the phenotype, as shown previously for wild-type AQP0. In the presence of calmidazolium, a CaM inhibitor, S235D showed high P(f) and no Ca(2+) regulation, suggesting that S235D still binds CaM. Contrarily, S229D showed a decrease in recruitment of CaM, suggesting that S229D is unable to bind CaM. Taken together, our results suggest a model in which CaM acts as an inhibitor of AQP0 P(f). CaM binding is associated with a low P(f) state, and a lack of CaM binding is associated with a high P(f) state. Pathological conditions of inappropriate phosphorylation or calcium/CaM regulation could induce P(f) changes contributing to the development of a cataract. PMID- 18508774 TI - Distinct transport mechanisms in yeast ammonium transport/sensor proteins of the Mep/Amt/Rh family and impact on filamentation. AB - Ammonium transport proteins of the Mep/Amt/Rh family include microbial and plant Mep/Amt members, crucial for ammonium scavenging, and animal Rhesus factors likely involved in ammonium disposal. Recent structural information on two bacterial Mep/Amt proteins has revealed the presence, in the hydrophobic conducting pore, of a pair of preserved histidines proposed to play an important role in substrate conductance, by participating either in NH(4)(+) deprotonation or in shaping the pore. Here we highlight the existence of two functional Mep/Amt subfamilies distinguishable according to whether the first of these histidines is conserved, as in yeast ScMep2, or replaced by glutamate, as in ScMep1. Replacement of the native histidine of ScMep2 with glutamate leads to conversion from ScMep2 to ScMep1-like properties. This includes a two-unit upshift of the optimal pH for transport and an increase of the transport rate, consistent with alleviation of an energy-limiting step. Similar effects are observed when the same substitution is introduced into the Escherichia coli AmtB protein. In contrast to ScMep1, ScMep2 is proposed to play an additional signaling role in the induction of filamentous growth, a dimorphic change often associated with virulence in pathogenic fungi. We show here that the histidine to glutamate substitution in ScMep2 leads to uncoupling of the transport and sensor functions, suggesting that a ScMep2-specific transport mechanism might be responsible for filamentation. Our overall data suggest the existence of two functional groups of Mep/Amt-type proteins with different transport mechanisms and distinct impacts on cell physiology and signaling. PMID- 18508775 TI - The N-terminal domain of annexin 2 serves as a secondary binding site during membrane bridging. AB - Annexin A2 (AnxA2) is a Ca(2+)- and acidic phospholipid-binding protein involved in many cellular processes. It undergoes Ca(2+)-mediated membrane bridging at neutral pH and has been demonstrated to be involved in an H(+)-mediated mechanism leading to a novel AnxA2-membrane complex structure. We used fluorescence techniques to characterize this H(+)-dependent mechanism at the molecular level; in particular, the involvement of the AnxA2 N-terminal domain. This domain was labeled at Cys-8 either with acrylodan or pyrene-maleimide fluorescent probes. Steady-state and time-resolved fluorescence analysis for acrylodan and fluorescence quenching by doxyl-labeled phospholipids revealed direct interaction between the N-terminal domain and the membrane. The absence of pyrene excimer suggested that interactions between N termini are not involved in the H(+) mediated mechanism. These findings differ from those previously observed for the Ca(2+)-mediated mechanism. Protein titration experiments showed that the protein concentration for half-maximal membrane aggregation was twice for Ca(2+)-mediated compared with H(+)-mediated aggregation, suggesting that AnxA2 was able to bridge membranes either as a dimer or as a monomer, respectively. An N-terminally deleted AnxA2 was 2-3 times less efficient than the wild-type protein for H(+) mediated membrane aggregation. We propose a model of AnxA2-membrane assemblies, highlighting the different roles of the N-terminal domain in the H(+)- and Ca(2+) mediated membrane bridging mechanisms. PMID- 18508776 TI - Cystic fibrosis transmembrane regulator missing the first four transmembrane segments increases wild type and DeltaF508 processing. AB - We previously generated an adenoassociated viral gene therapy vector, rAAV Delta264 cystic fibrosis transmembrane conductance regulator (CFTR), missing the first four transmembrane domains of CFTR. When infected into monkey lungs, Delta264 CFTR increased the levels of endogenous wild type CFTR protein. To understand this process, we transfected Delta264 CFTR plasmid cDNA into COS7 cells, and we noted that protein expression from the truncation mutant is barely detectable when compared with wild type or DeltaF508 CFTR. Delta264 CFTR protein expression increases dramatically when cells are treated with proteasome inhibitors. Cycloheximide experiments show that Delta264 CFTR is degraded faster than DeltaF508 CFTR. VCP and HDAC6, two proteins involved in retrograde translocation from endoplasmic reticulum to cytosol for proteasomal and aggresomal degradation, coimmunoprecipitate with Delta264 CFTR. In cotransfection studies in COS7 cells and in transfection of Delta264 CFTR into cells stably expressing wild type and DeltaF508 CFTR, Delta264 CFTR increases wild type CFTR protein and increases levels of maturation of immature band B to mature band C of DeltaF508 CFTR. Thus the adenoassociated viral vector, rAAV-Delta264 CFTR, is a highly promising cystic fibrosis gene therapy vector because it increases the amount of mature band C protein both from wild type and DeltaF508 CFTR and associates with key elements in quality control mechanism of CFTR. PMID- 18508777 TI - Antizyme inhibitor 2 (AZIN2/ODCp) stimulates polyamine uptake in mammalian cells. AB - One of the processes that regulate intracellular levels of polyamines in mammalian cells is polyamine uptake. We have measured polyamine uptake in COS7 cells for putrescine, spermidine, and spermine, obtaining K(m) values of 4.5, 1.0, and 0.8 mum, respectively. Treatment of nonconfluent cells with cycloheximide stimulated polyamine uptake and prevented the inhibitory effect found in cells preloaded with polyamines, suggesting the existence of a feedback repression mechanism mediated by antizymes. Transient transfected cells with mutated antizyme forms of AZ1, AZ2, and AZ3, which do not require frameshifting, showed a total blockade of polyamine uptake. Transfection of COS7 cells with mouse or human AZIN2, a novel member of the antizyme inhibitor family, recently characterized by our group, markedly stimulated polyamine uptake and counteracted the action of any of the three antizymes in co-transfected cells. The stimulatory effect of AZIN2 on polyamine uptake was abrogated when the putative antizyme binding sequence, formed by residues 117-140 in AZIN2, was deleted. Real time reverse transcription-PCR analysis of antizyme inhibitor transcripts revealed that in brain and testes AZIN2 is more expressed than AZIN1, especially in the testes where the relative expression was about 25-fold higher. Collectively, our results clearly indicate that AZIN2 affects polyamine homeostasis not only by increasing ornithine decarboxylase activity but also by stimulating polyamine uptake, through negating the inhibitory effect of the antizymes. This finding may have physiological relevance, mostly in testes where AZ3 and AZIN2 are mainly expressed. PMID- 18508778 TI - Analysis of the synaptotagmin family during reconstituted membrane fusion. Uncovering a class of inhibitory isoforms. AB - Ca(2+)-triggered exocytosis in neurons and neuroendocrine cells is regulated by the Ca(2+)-binding protein synaptotagmin (syt) I. Sixteen additional isoforms of syt have been identified, but little is known concerning their biochemical or functional properties. Here, we assessed the abilities of fourteen syt isoforms to directly regulate SNARE (soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor)-catalyzed membrane fusion. One group of isoforms stimulated neuronal SNARE-mediated fusion in response to Ca(2+), while another set inhibited SNARE catalyzed fusion in both the absence and presence of Ca(2+). Biochemical analysis revealed a strong correlation between the ability of syt isoforms to bind 1,2-dioleoyl phosphatidylserine (PS) and t-SNAREs in a Ca(2+) promoted manner with their abilities to enhance fusion, further establishing PS and SNAREs as critical effectors for syt action. The ability of syt I to efficiently stimulate fusion was specific for certain SNARE pairs, suggesting that syts might contribute to the specificity of intracellular membrane fusion reactions. Finally, a subset of inhibitory syts down-regulated the ability of syt I to activate fusion, demonstrating that syt isoforms can modulate the function of each other. PMID- 18508779 TI - Cellular differentiation in moss protonemata: a morphological and experimental study. AB - BACKGROUND AND AIMS: Previous studies of protonemal morphogenesis in mosses have focused on the cytoskeletal basis of tip growth and the production of asexual propagules. This study provides the first comprehensive description of the differentiation of caulonemata and rhizoids, which share the same cytology, and the roles of the cytoskeleton in organelle shaping and spatial arrangement. METHODS: Light and electron microscope observations were carried out on in vitro cultured and wild protonemata from over 200 moss species. Oryzalin and cytochalasin D were used to investigate the role of the cytoskeleton in the cytological organization of fully differentiated protonemal cells; time-lapse photography was employed to monitor organelle positions. KEY RESULTS: The onset of differentiation in initially highly vacuolate subapical cells is marked by the appearance of tubular endoplasmic reticulum (ER) profiles with crystalline inclusions, closely followed by an increase in rough endoplasmic reticulum (RER). The tonoplast disintegrates and the original vacuole is replaced by a population of vesicles and small vacuoles originating de novo from RER. The cytoplasm then becomes distributed throughout the cell lumen, an event closely followed by the appearance of endoplasmic microtubules (MTs) in association with sheets of ER, stacks of vesicles that subsequently disperse, elongate mitochondria and chloroplasts and long tubular extensions at both poles of the nucleus. The production of large vesicles by previously inactive dictysomes coincides with the deposition of additional cell wall layers. At maturity, the numbers of endoplasmic microtubules decline, dictyosomes become inactive and the ER is predominantly smooth. Fully developed cells remain largely unaffected by cytochalasin; oryzalin elicits profound cytological changes. Both inhibitors elicit the formation of giant plastids. The plastids and other organelles in fully developed cells are largely stationary. CONCLUSIONS: Differentiation of caulonemata and rhizoids involves a remarkable series of cytological changes, some of which closely recall major events in sieve element ontogeny in tracheophytes. The cytology of fully differentiated cells is remarkably similar to that of moss food-conducting cells and, in both, is dependent on an intact microtubule cytoskeleton. The disappearance of the major vacuolar apparatus is probably related to the function of caulonema and rhizoids in solute transport. Failure of fully differentiated caulonema and rhizoid cells to regenerate is attributed to a combination of endo-reduplication and irreversible tonoplast fragmentation. The formation of giant plastids, most likely by fusion, following both oryzalin and cytochalasin treatments, suggests key roles for both microtubules and microfilaments in the spatial arrangement and replication of plastids. PMID- 18508780 TI - A new LH receptor splice mutation responsible for male hypogonadism with subnormal sperm production in the propositus, and infertility with regular cycles in an affected sister. AB - BACKGROUND: Inactivating LH receptor (LHR) mutations have been described so far in men as well as in women. Phenotypes in men have been variable with in nearly all cases impairment of sex differentiation or azoospermia. We report a milder reproductive phenotype both in a male patient and his sister. METHODS AND RESULTS: We describe a family that carries a homozygous mutation G-->A at position -1 at the intron 10-exon 11 boundary of the LHR gene. The male patient presented with delayed puberty, micropenis and oligospermia. Two of his sisters were homozygous for the same mutation and were infertile. Surprisingly, one of them was found to have had regular ovarian cycles for years and showed normal LH values (6.5 and 10.6 mIU/ml for LH and FSH, respectively). In vitro analysis showed that this altered splicing resulted in an LHR from which eight amino acids are deleted from the extracellular domain (Delta Tyr(317)-Ser(324)). In vitro expression has shown that the receptor was expressed and capable of LH-induced signaling, albeit with reduced potency (P < 0.001). CONCLUSIONS: LHR mutations may represent an underestimated cause of infertility in women, in addition to being responsible for male hypogonadism with reduced spermatogenesis. PMID- 18508782 TI - Sudden arrhythmic death syndrome: familial evaluation identifies inheritable heart disease in the majority of families. AB - AIMS: At least 4% of sudden deaths are unexplained at autopsy [sudden arrhythmic death syndrome (SADS)] and a quarter may be due to inherited cardiac disease. We hypothesized that comprehensive clinical investigation of SADS families would identify more susceptible individuals and causes of death. METHODS AND RESULTS: Fifty seven consecutively referred families with SADS death underwent evaluation including resting 12 lead, 24 h and exercise ECG and 2D echocardiography. Other investigations included signal averaged ECG, ajmaline testing, cardiac magnetic resonance imaging, and mutation analysis. First-degree relatives [184/262 (70%)] underwent evaluation, 13 (7%) reporting unexplained syncope. Seventeen (30%) families had a history of additional unexplained premature sudden death(s). Thirty families (53%) were diagnosed with inheritable heart disease: 13 definite long QT syndrome (LQTS), three possible/probable LQTS, five Brugada syndrome, five arrhythmogenic right ventricular cardiomyopathy (ARVC), and four other cardiomyopathies. One SCN5A and four KCNH2 mutations (38%) were identified in 13 definite LQTS families, one SCN5A mutation (20%) in five Brugada syndrome families and one (25%) PKP2 (plakophyllin2) mutation in four ARVC families. CONCLUSION: Over half of SADS deaths were likely to be due to inherited heart disease; accurate identification is vital for appropriate prophylaxis amongst relatives who should undergo comprehensive cardiological evaluation, guided and confirmed by mutation analysis. PMID- 18508783 TI - Dislodgement of an Amplatzer occluder device causing iatrogenic pulmonary embolism in a patient with post-infarction ventricular septal defect. PMID- 18508784 TI - Short-latency median-nerve somatosensory-evoked potentials and induced gamma oscillations in humans. AB - Recent studies have suggested that cortical gamma-oscillations are tightly linked with various forms of physiological activity. In the present study, the dynamic changes of intracranially recorded median-nerve somatosensory-evoked potentials (SEPs) and somatosensory-induced gamma-oscillations were animated on a three dimensional MR image, and the temporal and spatial characteristics of these activities were analysed in 10 children being evaluated for epilepsy surgery. Visual and quantitative assessments revealed that short-latency SEPs and somatosensory-induced gamma-oscillations predominantly involved the post-central gyrus and less intensely involved the pre-central gyrus and the anterior parietal lobule. Formation of a dipole of N20 peak with opposite polarities across the central sulcus was well delineated in animation movies. High-frequency (100-250 Hz) somatosensory-induced gamma-oscillations emerged in the post-central gyrus at 13.6-17.5 ms after median-nerve stimulation, gradually slowed down in frequency around and below 100 Hz, and progressively involved the neighbouring areas. A substantial proportion of somatosensory-induced gamma-oscillations was initially phase-locked and the proportion of a non-phase-locked component gradually increased over time. The primary motor hand areas proven by cortical stimulation frequently coincided with the sites showing the largest N20 peak and the largest somatosensory-induced gamma oscillations. In vivo animation of SEPs and somatosensory-induced gamma oscillations both may be utilized to localize the primary sensory-motor hand area in pre-surgical evaluation. The dipole on SEPs is consistent with the previously accepted notion that the cortices along the central sulcus are activated. The high-frequency somatosensory-induced gamma oscillations in the post-central gyrus may represent the initial neural processing for external somatosensory stimuli, whereas the subsequent lower frequency oscillations might represent the reafferent cortical activity occurring in larger cortical networks. PMID- 18508785 TI - Are three additional cycles of chemotherapy useful in patients with advanced stage epithelial ovarian cancer after a complete response to six cycles of intravenous adjuvant paclitaxel and carboplatin? AB - BACKGROUND: To evaluate the efficacy of three additional cycles of chemotherapy in patients with the International Federation of Gynecology and Obstetrics Stage III or IV, who achieved a complete response after six cycles of intravenous adjuvant paclitaxel/carboplatin after surgery. METHODS: The clinical data of 94 patients with complete response after six cycles of adjuvant paclitaxel/carboplatin after surgery between January 1997 and March 2007 were reviewed retrospectively. Three additional cycles using the same chemotherapy were administered to 57 patients as consolidation chemotherapy (Group 1). Thirty seven patients without the additional cycles served as controls (Group 2). Disease-free survival (DFS) and overall survival (OS) were evaluated using the Kaplan-Meier method with the log-rank test. The importance of consolidation chemotherapy as a prognostic factor affecting survival was examined using the Cox's proportional hazard analysis. The incidence of chemotherapy-induced hematological toxicities was compared between the two groups using chi-square test. RESULTS: Median DFS and mean OS were not significantly different between the two groups (15 versus 22 months, P = 0.703; 69 versus 73 months, P = 0.891, respectively). Consolidation chemotherapy was not a prognostic factor of survival although optimal debulking surgery and lower value of serum CA-125 levels after six cycles of the chemotherapy were prognostic factors improving DFS (P < 0.01). Grade 3 or 4 leukopenia was more common in patients treated with consolidation chemotherapy than in those not treated (50.9 versus 21.6%, P = 0.004). CONCLUSION: Consolidation chemotherapy using paclitaxel/carboplatin may be inefficient and relatively toxic to advanced-stage epithelial ovarian cancer patients with complete response to six cycles of the same chemotherapy after surgery. PMID- 18508786 TI - First case of a primary osteosarcoma of the ureter: diagnostic findings, course of disease and treatment. PMID- 18508787 TI - A novel heterozygous HIF2AM535I mutation reinforces the role of oxygen sensing pathway disturbances in the pathogenesis of familial erythrocytosis. AB - HIF2A transcription factor plays a central role in the regulation of the hypoxia responding pathway in mammalian cells, by modulating erythropoiesis and angiogenesis. Molecular alterations of oxygen sensing pathway constituents are implicated in hereditary erythrocytosis. Here we show that 2 members of a family with idiopathic erythrocytosis exhibited a new heterozygous G to A mutation at base 1605 of the exon 12 of hypoxia-inducible factor-2A (HIF2A) gene. This mutation determines the replacement of methionine by isoleucine at the position 535, very close to the position 531, where the hydroxyl acceptor prolyne is located. In addition, we found that mRNA expression of erythropoietin receptor, vascular endothelial growth factor, transferrin receptor, adrenomedullin and N myc downstream regulated gene 1, up-regulated by HIF2A or hypoxia, were significantly higher in patients carrying the mutation than in normal controls. These results suggest that the HIF2A(M535I) gene mutation could induce hereditary erythrocytosis at a young age. PMID- 18508788 TI - A possible role for low-dose cyclosporine in refractory immune thrombocytopenic purpura. PMID- 18508789 TI - The MDM2 -309 T/G promoter single nucleotide polymorphism does not alter disease characteristics in chronic lymphocytic leukemia. PMID- 18508790 TI - Aberrant expression of microRNA in polycythemia vera. AB - BACKGROUND: Polycythemia vera is a clonal hematopoietic stem cell disorder in which the JAK2 V617F mutation is observed in >95% of patients, but an as yet unidentified process appears to initiate the clonal expansion of hematopoiesis. Because microRNA regulate hematopoietic differentiation, we hypothesized that dysregulated expression of microRNA may contribute to the pathophysiology of polycythemia vera. DESIGN AND METHODS: We performed gene expression profiling in five patients with polycythemia vera and in five controls using CombiMatrix MicroRNA CustomArray. ANOVA identified deregulated microRNA in polycythemia vera, and their expression was studied in a larger set of samples by quantitative reverse transcriptase polymerase chain reaction. The expression of these microRNA was also analyzed in other myeloproliferative disorders. RESULTS: We observed down-regulation of let-7a and up-regulation of miR-182 in polycythemia vera granulocytes, up-regulation of miR-143, miR-145 and miR-223 in polycythemia vera mononuclear cells, up-regulation of miR-26b in polycythemia vera platelets, and down-regulation of miR-30b, miR-30c and miR-150 in polycythemia vera reticulocytes. JAK2 V617F frequency was positively correlated with miR-143 expression and inversely correlated with let-7a, miR-30c, miR-342 and miR-150. Transcript level of predicted target genes was determined, and overexpression of IRAK2 was detected in all granulocytes from patients with myeloproliferative disorders and in polycythemia vera reticulocytes. Abnormally high HMGA2 microRNA was found in myelofibrosis granulocytes. CONCLUSIONS: Our study demonstrates that peripheral blood cells from patients with polycythemia vera have microRNA signatures distinct from those of controls. Our findings of aberrant microRNA expression underline the complexity of the molecular basis of polycythemia vera. PMID- 18508791 TI - Genome-wide analysis of copy number changes and loss of heterozygosity in myelodysplastic syndrome with del(5q) using high-density single nucleotide polymorphism arrays. AB - BACKGROUND: We undertook a genome wide single nucleotide polymorphism analysis of a spectrum of patients with myelodysplastic syndrome del(5q) in order to investigate whether additional genomic abnormalities occur. Single nucleotide polymorphism array analysis has been shown to detect not only gene deletions but also regions of uniparental disomy that can pinpoint particular regions for mutation analysis. DESIGN AND METHODS: We studied 42 cases of myelodysplastic syndrome with del(5q), comprising 21 patients with 5q- syndrome and 21 with del(5q) (not 5q- syndrome), and 45 healthy controls by genome wide single nucleotide polymorphism analysis with the 50K Affymetrix single nucleotide polymorphism arrays. RESULTS: The del(5q) was characterized in all cases. The commonly deleted region of the 5q- syndrome extends between the genes SH3TC2 (proximal boundary) and GLRA1 (distal boundary) and measures 2.9 Mb. Copy number changes in addition to the del(5q) were observed in 10 of 21 patients with del(5q) myelodysplastic syndrome but in none of the patients with the 5q- syndrome. A total of 63 regions of uniparental disomy greater than 2 Mb were detected in 40 of 42 patients, dispersed on 18/23 chromosomes. In the 5q- syndrome group 31 regions of uniparental disomy were identified in 19 of 21 patients, the largest one being 7.6 Mb. All 21 patients with del(5q) myelodysplastic syndrome had uniparental disomy; in total 32 regions of uniparental disomy were identified in the 21 patients, including six regions of uniparental disomy > 10 Mb. Eight recurrent regions of uniparental disomy were observed among the 42 patients. For eight patients we had T-cell DNA as a germline control and four recurrent regions of uniparental disomy were identified that were present only in the neutrophil and not T-cell DNA. One small region of uniparental disomy at 10p12.31-p12.2 was observed in four patients with the 5q- syndrome. CONCLUSIONS: This study shows that regions of uniparental disomy greater than 2 Mb are found in the 5q-syndrome and del(5q) myelodysplastic syndrome, although large regions of uniparental disomy (>10 Mb) are only found in the latter group. The recurrent regions of uniparental disomy may indicate the position of novel leukemia-associated genes. PMID- 18508792 TI - IgM anti-protein S antibodies as a risk factor for venous thrombosis. PMID- 18508793 TI - The IL6(-174G/C) polymorphism is a prognostic factor for survival after treatment initiation in Waldenstrom macroglobulinemia patients aged 65 years or less. PMID- 18508794 TI - Granulocyte concentrates: prolonged functional capacity during storage in the presence of phenotypic changes. AB - BACKGROUND: Granulocyte transfusion has been proposed as a bridging therapy for patients with prolonged periods of chemotherapy-induced neutropenia, who suffer from severe fungal and bacterial infections. To recover, adequate numbers of granulocytes are required when the patients are refractory to standard treatment. The aim of this study was to assess the functional characteristics and efficacy of granulocyte colony-stimulating factor/dexamethasone-mobilized granulocytes used for transfusions. DESIGN AND METHODS: Granulocytes from the leukapheresis products were tested for the expression of cell-surface antigens, interactions with endothelial cells, motility, killing of microbes and survival. The granulocytes were used in vivo for transfusion in 16 severely ill children, who were--apart from a patient with a granulocyte dysfunction--all suffering from prolonged neutropenia. RESULTS: Mobilization of granulocytes with granulocyte colony-stimulating factor and dexamethasone caused phenotypic changes (decreased CD62L expression and increased levels of CD66b and CD177). The ability of the granulocytes to interact with endothelial cells (rolling, adhesion, transmigration) and to kill various types of pathogens was not affected by the mobilization, leukapheresis and irradiation procedures. The granulocytes were functionally indistinguishable from those isolated from untreated donors, even after 24 hours of storage. Granulocyte transfusion seemed to benefit 70% of patients, as 11 out of the 16 children showed clinical recovery within 1-2 weeks after beginning the transfusions. CONCLUSIONS: Although CD62L expression is downregulated on granulocytes used for granulocyte transfusions, concomitant CD177 upregulation may explain the intact interactions with endothelial cells. All other granulocyte functions tested were intact, including the ability to kill fungi. Granulocyte concentrates can be stored without loss of in vitro viability and functionality for at least 24 hours. As demonstrated in vivo, granulocyte transfusions may be an effective therapy for neutropenic pediatric patients suffering from life-threatening infections. PMID- 18508795 TI - No evidence of cardiac iron in 20 never- or minimally-transfused patients with thalassemia intermedia. PMID- 18508796 TI - Low-dose thalidomide in myelofibrosis. PMID- 18508797 TI - Increased cortical bone mineralization in imatinib treated patients with chronic myelogenous leukemia. PMID- 18508798 TI - RETRACTED: Heterogeneous promoter activity of the telomerase reverse transcriptase gene in individual acute myeloid leukemia cells defined by lentiviral reporter assay. PMID- 18508799 TI - Prognostic impact of genetic characterization in the GIMEMA LAM99P multicenter study for newly diagnosed acute myeloid leukemia. AB - BACKGROUND: Recent advances in genetic characterization of acute myeloid leukemia indicate that combined cytogenetic and molecular analyses provide better definition of prognostic groups. The aim of this study was to verify this prospectively in a large group of patients. DESIGN AND METHODS: Genetic characterization was prospectively carried out in 397 patients with acute myeloid leukemia (median age, 46 years) receiving uniform treatment according to the LAM99P protocol of the Italian GIMEMA group. The impact of genetic markers on response to therapy and outcome was assessed by univariate and multivariate analyses. RESULTS: For induction response, conventional karyotyping identified three groups with complete remission rates of 92%, 67% and 39% (p<0.0001). Complete remission rates in NPM1 mutated (NPM1+) and wild-type (NPM1-) groups were 76% and 60%, respectively, for the whole population and 81% and 61% in the group with normal karyotype (p<0.001 and p=0.026, respectively). Multivariate analysis indicated that low risk karyotype and NPM1+ were independent factors favorably affecting complete remission. Multivariate analysis of overall and disease-free survival among 269 patients who achieved complete remission showed a significant impact of karyotype on both estimates and of FLT3 status on disease free-survival (FLT3-ITD vs. FLT3 wild-type, p=0.0001). NPM1 status did not significantly influence disease free-survival in either the whole population or in the patients with a normal karyotype in this series, probably due to the low number of cases analyzed. CONCLUSIONS: These results reiterate the prognostic relevance of combining cytogenetic and mutational analysis in the diagnostic work up of patients with acute myeloid leukemia. PMID- 18508800 TI - Expression of the nuclear oncogene Ski in patients with acute myeloid leukemia treated with all-trans retinoic acid. PMID- 18508801 TI - M4 acute myeloid leukemia: the role of eosinophilia and cytogenetics in treatment response and survival. The GIMEMA experience. AB - BACKGROUND: Myelomonocytic acute myeloid leukemia (M4-AML) is frequently associated with the cytogenetic marker inv(16) and/or the presence of eosinophilia. The aim of this study was to analyze the incidence and prognostic role of these factors in a large series of patients. DESIGN AND METHODS: Adult patients with acute myeloid leukemia consecutively enrolled in the GIMEMA trials AML10 and LAM99p were retrospectively analyzed. RESULTS: Among 1686 patients, 400 cases of M4-AML were identified; of these, 78% had neither eosinophilia nor inv(16), 6% had eosinophilia only, 8% had inv(16) only and 8% had both. Univariate analysis showed that both eosinophilia and inv(16) were correlated with a higher probability of complete remission, lower resistance to chemotherapy and increased overall survival. Multivariate analysis showed that the simultaneous presence of the two factors significantly increased the probabilities of both complete remission and overall survival. The presence of only one of the two factors also increased the probabilities of complete remission and overall survival, but not to a statistically significant extent. The relapse-free survival of the responding patients was not influenced by the two factors. CONCLUSIONS: In a large series of patients with M4-AML we confirmed the favorable role of inv(16), but the weight of this factor among the whole M4 population was of limited relevance. Eosinophilia, which affects a small proportion of cases, also emerged as a favorable prognostic factor. Based on the results of this large case population, overall and relapse-free survival rates of patients with M4-AML are not significantly better than those of patients with non M4 AML, while the concomitant presence of both inv(16) and eosinophilia was associated with a significantly improved prognosis. PMID- 18508802 TI - NOTCH2 mutations in marginal zone lymphoma. PMID- 18508803 TI - Influence of sickle cell disease and treatment with hydroxyurea on sperm parameters and fertility of human males. AB - BACKGROUND: Recent progress in the treatment of sickle cell disease, in particular the use of hydroxyurea, has considerably modified the prognosis of this disease. Many more patients now reach reproductive age. The objective of this study was to assess the potential impact of hydroxyurea on the semen of patients. DESIGN AND METHODS: In this retrospective multicenter study, we evaluated the sperm parameters and fertility of 44 patients and analyzed the potential impact of hydroxyurea. RESULTS: We report data from the largest series so far of semen analyses in patients with sickle cell disease: 108 samples were analyzed, of which 76 were collected before treatment. We found that at least one sperm parameter was abnormal in 91% of the patients before treatment, in agreement with published literature. All sperm parameters seemed to be affected in semen samples collected during hydroxyurea treatment, and this impairment occurred in less than 6 months, later reaching a plateau. Furthermore, after hydroxyurea cessation, while global results in 30 patients were not statistically different before and after hydroxyurea treatment, in four individuals follow-up sperm parameters did not seem to recover quickly and the total number of spermatozoa per ejaculate fell below the normal range in about half the cases. CONCLUSIONS: The observed alterations of semen parameters due to sickle cell disease seem to be exacerbated by hydroxyurea treatment. Until prospective studies reveal reassuring findings, we suggest that a pre-treatment sperm analysis be performed and sperm cryopreservation be offered to patients before hydroxyurea treatment. PMID- 18508804 TI - Enzymatic and metabolic diagnostic of nitrogen deficiency in Arabidopsis thaliana Wassileskija accession. AB - Adaptation to steady-state low-nutrient availability was investigated by comparing the Wassileskija (WS) accession of Arabidopsis thaliana grown on 2 or 10 mM nitrate. Low nitrogen conditions led to a limited rosette biomass and seed yield. The latter was mainly due to reduced seed number, while seed weight was less affected. However, harvest index was lower in high nitrate compared with limited nitrate conditions. Under nitrogen-limiting conditions, nitrate reductase activity was decreased while glutamine synthetase activity was increased due to a higher accumulation of the cytosolic enzyme. The level of nitrogen remobilization to the seeds was higher under low nitrogen, and the vegetative parts of the plants remaining after seed production stored very low residual nitrogen. Through promoting nitrogen remobilization and recycling pathways, nitrogen limitation modified plant and seed compositions. Rosette leaves contained more sugars and less free amino acids when grown under nitrogen-limiting conditions. Compared with high nitrogen, the levels of proline, asparagine and glutamine were decreased. The seed amino acid composition reflected that of the rosette leaves, thus suggesting that phloem loading for seed filling was poorly selective. The major finding of this report was that together with decreasing biomass and yield, nitrogen limitation triggers large modifications in vegetative products and seed quality. PMID- 18508805 TI - A single amino acid change in histone H4 enhances UV survival and DNA repair in yeast. AB - Single amino acid changes at specific DNA contacts of histones H3 and H4 generate SWI/SNF-independent (Sin) mutants in yeast. We have analyzed the effect of the Sin mutation at R45 of histone H4 on cell survival following UV irradiation, nucleotide excision repair (NER) and chromatin structure. We find that this mutation renders yeast cells more resistant to UV damage and enhances NER at specific chromatin loci. In the transcriptionally silent HML, repressed GAL10 and the constitutively active RPB2 loci, H4 R45 mutants exhibit enhanced repair of UV induced cyclobutane pyrimidine dimers (CPDs) compared to wild-type (wt). However, the H4 R45 mutation does not increase the transcription of NER genes, disrupt transcriptional silencing of the HML locus or alter repression in the GAL10 locus. We have further shown that the H4 R45C mutation increases the accessibility of nucleosome DNA in chromatin to exogenous nucleases and may expedite nucleosome rearrangements during NER. Taken together, our results indicate that the increased repair observed in Sin mutants is a direct effect of the altered chromatin landscape caused by the mutation, suggesting that such subtle changes in the conserved histone residues can influence the accessibility of DNA repair factors in chromatin. PMID- 18508806 TI - GEPAS, a web-based tool for microarray data analysis and interpretation. AB - Gene Expression Profile Analysis Suite (GEPAS) is one of the most complete and extensively used web-based packages for microarray data analysis. During its more than 5 years of activity it has continuously been updated to keep pace with the state-of-the-art in the changing microarray data analysis arena. GEPAS offers diverse analysis options that include well established as well as novel algorithms for normalization, gene selection, class prediction, clustering and functional profiling of the experiment. New options for time-course (or dose response) experiments, microarray-based class prediction, new clustering methods and new tests for differential expression have been included. The new pipeliner module allows automating the execution of sequential analysis steps by means of a simple but powerful graphic interface. An extensive re-engineering of GEPAS has been carried out which includes the use of web services and Web 2.0 technology features, a new user interface with persistent sessions and a new extended database of gene identifiers. GEPAS is nowadays the most quoted web tool in its field and it is extensively used by researchers of many countries and its records indicate an average usage rate of 500 experiments per day. GEPAS, is available at http://www.gepas.org. PMID- 18508807 TI - ENDEAVOUR update: a web resource for gene prioritization in multiple species. AB - Endeavour (http://www.esat.kuleuven.be/endeavourweb; this web site is free and open to all users and there is no login requirement) is a web resource for the prioritization of candidate genes. Using a training set of genes known to be involved in a biological process of interest, our approach consists of (i) inferring several models (based on various genomic data sources), (ii) applying each model to the candidate genes to rank those candidates against the profile of the known genes and (iii) merging the several rankings into a global ranking of the candidate genes. In the present article, we describe the latest developments of Endeavour. First, we provide a web-based user interface, besides our Java client, to make Endeavour more universally accessible. Second, we support multiple species: in addition to Homo sapiens, we now provide gene prioritization for three major model organisms: Mus musculus, Rattus norvegicus and Caenorhabditis elegans. Third, Endeavour makes use of additional data sources and is now including numerous databases: ontologies and annotations, protein-protein interactions, cis-regulatory information, gene expression data sets, sequence information and text-mining data. We tested the novel version of Endeavour on 32 recent disease gene associations from the literature. Additionally, we describe a number of recent independent studies that made use of Endeavour to prioritize candidate genes for obesity and Type II diabetes, cleft lip and cleft palate, and pulmonary fibrosis. PMID- 18508808 TI - PBEQ-Solver for online visualization of electrostatic potential of biomolecules. AB - PBEQ-Solver provides a web-based graphical user interface to read biomolecular structures, solve the Poisson-Boltzmann (PB) equations and interactively visualize the electrostatic potential. PBEQ-Solver calculates (i) electrostatic potential and solvation free energy, (ii) protein-protein (DNA or RNA) electrostatic interaction energy and (iii) pKa of a selected titratable residue. All the calculations can be performed in both aqueous solvent and membrane environments (with a cylindrical pore in the case of membrane). PBEQ-Solver uses the PBEQ module in the biomolecular simulation program CHARMM to solve the finite difference PB equation of molecules specified by users. Users can interactively inspect the calculated electrostatic potential on the solvent-accessible surface as well as iso-electrostatic potential contours using a novel online visualization tool based on MarvinSpace molecular visualization software, a Java applet integrated within CHARMM-GUI (http://www.charmm-gui.org). To reduce the computational time on the server, and to increase the efficiency in visualization, all the PB calculations are performed with coarse grid spacing (1.5 A before and 1 A after focusing). PBEQ-Solver suggests various physical parameters for PB calculations and users can modify them if necessary. PBEQ Solver is available at http://www.charmm-gui.org/input/pbeqsolver. PMID- 18508809 TI - PIE: an online prediction system for protein-protein interactions from text. AB - Protein-protein interaction (PPI) extraction has been an important research topic in bio-text mining area, since the PPI information is critical for understanding biological processes. However, there are very few open systems available on the Web and most of the systems focus on keyword searching based on predefined PPIs. PIE (Protein Interaction information Extraction system) is a configurable Web service to extract PPIs from literature, including user-provided papers as well as PubMed articles. After providing abstracts or papers, the prediction results are displayed in an easily readable form with essential, yet compact features. The PIE interface supports more features such as PDF file extraction, PubMed search tool and network communication, which are useful for biologists and bio system developers. The PIE system utilizes natural language processing techniques and machine learning methodologies to predict PPI sentences, which results in high precision performance for Web users. PIE is freely available at http://bi.snu.ac.kr/pie/. PMID- 18508810 TI - Sugars and flowering in the grapevine (Vitis vinifera L.). AB - Sugars play an important role in grapevine flowering. This complex process from inflorescence initiation to fruit maturity takes two growing seasons. Currently, most of the available data concern the involvement of sugars as energy sources during the formation of reproductive structures from initiation of inflorescences during the summer of the first year, until flower opening during the following spring. Sugars devoted to the development of reproductive structures are supplied either by wood reserves or by photosynthesis in leaves or inflorescences, depending on the stage of development. Female meiosis appears to be a key point in the success of flower formation because (i) flowers are vulnerable at this stage and (ii) it corresponds in the whole plant to the transition between reserve mobilization from perennial organs (roots, trunk, and canes) towards efficient leaf photosynthesis. The perturbation of reserve replenishment during the previous year provokes perturbation in the development of inflorescences, whereas altering the photosynthetic sources affects the formation of flowers during the same year. In particular, a lack of sugar availability in flowers at female meiosis caused by various environmental or physiological fluctuations may lead to drastic flower abortion. Apart from energy, sugars also play roles as regulators of gene expression and as signal molecules that may be involved in stress responses. In the future, these two topics should be further investigated in the grapevine considering the sensitivity of flowers to environmental stresses at meiosis. PMID- 18508811 TI - Quantitative expression analysis of selected COR genes reveals their differential expression in leaf and crown tissues of wheat (Triticum aestivum L.) during an extended low temperature acclimation regimen. AB - A number of COR genes (COld-Regulated genes) have been implicated in the acquisition of low temperature (LT) tolerance in wheat (Triticum aestivum L.). This study compared the relative expression patterns of selected COR genes in leaf and crown tissues of wheat near-isogenic lines to increase understanding of the molecular mechanisms underlying LT acclimation. Reciprocal near-isogenic lines were generated such that the dominant Vrn-A1 and recessive vrn-A1 loci were interchanged in a spring cv. Manitou and a winter cv. Norstar. Phenological development, acquisition of LT tolerance, and WCS120 polypeptide accumulation in these genotypes proceeded at rates similar to those previously reported for 6 degrees C acclimation from 0 to 98 d. However, a differential accumulation of WCS120 polypeptide and expression of the COR genes Wcs120, Wcor410, and Wcor14 was observed in the leaf and crown tissues. COR gene transcript levels peaked at 2 d of the acclimation period in both tissues and differences among genotypes were most evident at this time. COR gene expression was highest for the LT tolerant and lowest for the tender genotypes. However, expression rates were divergent enough in genotypes with intermediate hardiness that comparisons among tissues and/or times during acclimation often resulted in variable interpretations of the relative expression of the COR genes in the determination of LT tolerance. These observations emphasize the need to pay close attention to experimental conditions, sampling times, and genotype and tissue selection in experiments designed to identify the critical genetic components that interact to determine LT acclimation. PMID- 18508812 TI - Concerted modulation of alanine and glutamate metabolism in young Medicago truncatula seedlings under hypoxic stress. AB - The modulation of primary nitrogen metabolism by hypoxic stress was studied in young Medicago truncatula seedlings. Hypoxic seedlings were characterized by the up-regulation of glutamate dehydrogenase 1 (GDH1) and mitochondrial alanine aminotransferase (mAlaAT), and down-regulation of glutamine synthetase 1b (GS1b), NADH-glutamate synthase (NADH-GOGAT), glutamate dehydrogenase 3 (GDH3), and isocitrate dehydrogenase (ICDH) gene expression. Hypoxic stress severely inhibited GS activity and stimulated NADH-GOGAT activity. GDH activity was lower in hypoxic seedlings than in the control, however, under either normoxia or hypoxia, the in vivo activity was directed towards glutamate deamination. (15)NH(4) labelling showed for the first time that the adaptive reaction of the plant to hypoxia consisted of a concerted modulation of nitrogen flux through the pathways of both alanine and glutamate synthesis. In hypoxic seedlings, newly synthesized (15)N-alanine increased and accumulated as the major amino acid, asparagine synthesis was inhibited, while (15)N-glutamate was synthesized at a similar rate to that in the control. A discrepancy between the up-regulation of GDH1 expression and the down-regulation of GDH activity by hypoxic stress highlighted for the first time the complex regulation of this enzyme by hypoxia. Higher rates of glycolysis and ethanol fermentation are known to cause the fast depletion of sugar stores and carbon stress. It is proposed that the expression of GDH1 was stimulated by hypoxia-induced carbon stress, while the enzyme protein might be involved during post-hypoxic stress contributing to the regeneration of 2-oxoglutarate via the GDH shunt. PMID- 18508813 TI - Pharmaceutical reform in South Korea and the lessons it provides. AB - Through implementation of its 2000 pharmaceutical reform, the South Korean government expected to reduce the cost of medications and improve service levels, medical appropriateness of care, and drug effectiveness. However, despite the reform's lofty goals, unintended consequences have distorted the supply of medical services and spending. These consequences have included increasing the use of uninsured services, prescribing high-price drugs, and a growing market share for multinational drug companies. Further reforms are needed to reduce the measure's adverse effects. This paper examines the Korean mandatory prescription system and offers an analysis of Korea's reforms. PMID- 18508814 TI - Nutritional deficit as a negative prognostic factor in patients with miliary tuberculosis. AB - The effects of malnutrition on outcomes in miliary tuberculosis (MTB) are not well described. The aim of the present study was to find predictors for the development of acute respiratory failure (ARF) and survival in MTB patients, focusing on parameters reflecting nutritional condition. Out of the patients from three hospitals who had microbiologically or histopathologically confirmed tuberculosis, 56 patients presenting with typical disseminated pulmonary nodules on radiographs were retrospectively enrolled. A four-point nutritional risk score (NRS) was defined according to the presence of four nutritional factors: low body mass index (BMI; <18.5 kg x m(-2)), hypoalbuminaemia (<30.0 g x L(-1)), hypocholesterolaemia (<2.33 mmol x L(-1)) and severe lymphocytopenia (<7 x 10(5) cells x L(-1)). The male to female ratio was 1:3. ARF developed in 25% of patients (14 out of 56), with a 50% fatality rate. A high NRS (> or =3 points) was an independent risk factor for the development of ARF and fatality. In 90-day survival analysis, ARF, severe lymphocytopenia, hypocholesterolaemia, low BMI and higher NRS were risk factors for poor outcome. In multivariate analysis, only high NRS was an independent risk factor for 90-day survival rate in patients with MTB. A high nutritional risk score was a good predictor of poor outcome in miliary tuberculosis patients. Additional approaches to recover the nutritional deficits may become a focus in future management of miliary tuberculosis. PMID- 18508815 TI - Microsatellite DNA instability and COPD exacerbations. AB - Increased frequency of microsatellite DNA instability (MSI) has been detected in the sputum of chronic obstructive pulmonary disease (COPD) patients. The aim of the present study was to investigate the relationship between MSI in sputum cells and exacerbation frequency, which is an important parameter in the clinical course of the disease. Induced sputum samples and peripheral blood obtained from 36 patients with COPD at stable state were analysed. The control group consisted of 30 nonsmoking healthy subjects. DNA was extracted and analysed for MSI using the following microsatellite markers: RH70958, D5S207, D6S2223, D6S344, D6S263, G29802, D13S71, D14S588, D14S292 and D17S250. Following MSI analysis, exacerbations were recorded for 3 yrs in total. No MSI was detected in healthy nonsmokers. A total of 18 (50%) out of 36 patients exhibited MSI in their sputum cells. Patients who exhibited MSI showed significantly increased frequency of exacerbations compared with patients that did not. In addition, a significantly increased frequency of purulent and of severe type exacerbations was found in patients exhibiting MSI. Patients positive for marker G29802, D13S71 or D14S588 presented increased exacerbation frequency. The significant association between microsatellite DNA instability and chronic obstructive pulmonary disease exacerbations indicates that somatic mutations could be involved in the pathogenesis and natural history of the disease. PMID- 18508816 TI - Frequent epidermal growth factor receptor gene mutations in malignant pleural effusion of lung adenocarcinoma. AB - Malignant pleural effusions (MPEs) are often observed in lung cancer, especially adenocarcinoma. Epidermal growth factor receptor (EGFR) gene mutations are usually detected in lung adenocarcinoma. The purpose of the present study was to investigate the EGFR mutation rate in MPEs of lung adenocarcinoma. Between June 2005 and December 2006, 136 MPEs from lung adenocarcinoma were collected for EGFR mutation detection. In addition, between April 2001 and November 2004, 91 surgically resected specimens of lung adenocarcinoma from patients without MPEs were assessed for EGFR mutation. The EGFR mutation rate was significantly higher in the patients with MPEs than in the patients without (68.4% versus 50.5%). The EGFR mutation rate in patients with MPEs was not associated with sex, smoking history, age or cancer stage. By multivariate analysis, an age of <65 yrs, never smoking, Eastern Cooperative Oncology Group performance status 0-1, and EGFR mutation were significantly associated with a longer overall survival for lung adenocarcinoma patients with MPEs. The patients with malignant pleural effusions related to lung adenocarcinoma had a higher epidermal growth factor receptor gene mutation rate than the patients from whom surgically resected specimens were taken. Epidermal growth factor receptor tyrosine kinase inhibitors may be the treatment of choice for lung adenocarcinoma with malignant pleural effusions in east Asia. PMID- 18508817 TI - The lectin-like domain of thrombomodulin protects against ischaemia-reperfusion lung injury. AB - Ischaemia-reperfusion injury of the lung is a major cause of morbidity and mortality, particularly following lung transplantation, the mainstay treatment for patients with end-stage pulmonary disease. Effective measures to prevent this complication are lacking. Thrombomodulin (TM) is an endothelial cell receptor and cofactor for thrombin-mediated generation of the anticoagulant and anti inflammatory activated protein C (APC). The N-terminal lectin-like domain (LLD) of TM has no direct effects on coagulation, but has distinct anti-inflammatory properties, interfering with leukocyte adhesion, complement activation and cytokine generation, all of which are hallmarks of ischaemia-reperfusion injury. Using a murine model of lung ischaemia-reperfusion injury (90 min ischaemia, 4 h reperfusion), the present study shows that mice lacking the LLD of TM respond with increased extravasation of neutrophils and macrophages into the lung parenchyma and bronchoalveolar fluid (BALF), with augmented BALF levels of cytokines interleukin (IL)-1beta and granulocyte-monocytic colony-stimulating factor (GM-CSF). Pre-treatment of wild-type mice with recombinant LLD, as compared with controls, significantly suppresses protein leakage and accumulation of leukocytes in the BALF. These novel findings support further evaluation of recombinant lectin-like domain of thrombomodulin to protect the lung against tissue-damaging pro-inflammatory responses following ischaemia-reperfusion. PMID- 18508818 TI - Exhaled nitric oxide predicts lung function decline in difficult-to-treat asthma. AB - A subset of patients with asthma is known to have progressive loss of lung function despite treatment with corticosteroids. The aim of the present study was to identify risk factors of decline in forced expiratory volume in one second (FEV(1)) in patients with difficult-to-treat asthma. In total, 136 nonsmoking patients with difficult-to-treat asthma were recruited between 1998 and 1999. Follow-up assessment was performed 5-6 yrs later in 98 patients. The predictive effect of clinical characteristics and inflammatory markers were analysed at baseline (asthma onset and duration, atopy, airway hyperresponsiveness, blood and sputum eosinophils, and the fraction of nitric oxide in exhaled air (F(eNO))) on subsequent decline in post-bronchodilator FEV(1). Patients with high F(eNO) (> or =20 ppb) had an excess decline of 40.3 (95% confidence interval (CI) 7.3-73.2) mL.yr(-1) compared to patients with low F(eNO). F(eNO) > or =20 ppb was associated with a relative risk of 1.9 (95% CI, 1.1-2.6) of having an accelerated (> or =25 mL.yr(-1)) decline in FEV(1). In patients with baseline FEV(1) > or =80% of predicted, this relationship was even stronger: 90 versus 29% had accelerated decline in FEV(1) (F(eNO) > or =20 ppb versus F(eNO) <20 ppb respectively; relative risk 3.1 (95% CI, 1.7-3.4). Exhaled nitric oxide is a predictor of accelerated decline in lung function in patients with difficult-to treat asthma, particularly if forced expiratory volume in one second is still normal. PMID- 18508819 TI - Predictors of outcomes in COPD exacerbation cases presenting to the emergency department. AB - The aim of the present prospective multicentric study was to develop a simple rule for the prediction of poor outcome in patients presenting to emergency departments with initially non-life threatening-chronic obstructive pulmonary disease (COPD) exacerbations in a real-life setting. All patients with an acute exacerbation of COPD visiting the emergency departments of 103 hospitals during a 3-month period were included, except those who immediately required intensive care unit admission and/or ventilatory support. The data collected included patient characteristics, in-hospital outcomes (mortality and length of stay) and mode of discharge (unsupported or need for post-hospital assistance). The in hospital mortality rate was 7.4% (59 out of 794). Independent prognostic factors were age, number of clinical signs of severity (among cyanosis, impaired neurological status, lower limb oedema, asterixis and use of accessory inspiratory or expiratory muscles) and dyspnoea grade in the stable state. The need for post-hospital support was also predicted by female sex. In order to construct and validate a prediction score for mortality based on these items, patients were randomly allocated to a derivation and a validation cohort. The prediction score showed good discrimination, with a c-statistic of 0.79 in the derivation cohort and 0.83 in the validation cohort. Thus simple purely clinical factors can reliably predict the risk of death and requirement for post-hospital support in an initially non-life threatening-acute exacerbation of chronic obstructive pulmonary disease. Their use needs to be prospectively validated. PMID- 18508820 TI - Early mortality in patients with community-acquired pneumonia: causes and risk factors. AB - The first 48 h of evolution of patients with community-acquired pneumonia (CAP) are critical. The aim of the present study was to determine the frequency, causes and factors associated with early mortality in CAP. Nonimmunocompromised adults hospitalised with CAP were prospectively observed from 1995 to 2005. Early deaths, defined as death due to any cause < or = 48 h after admission, were compared with all patients who survived > 48 h. Furthermore, early deaths were compared with late deaths (patients who died > 48 h) and with survivors. Of 2,457 patients, 57 (2.3%) died < or = 48 h after admission. Overall mortality was 7.7%. The main causes of early mortality were respiratory failure and septic shock/multiorgan failure. Independent factors associated with early deaths were increased age, altered mental status at presentation, multilobar pneumonia, shock at admission, pneumococcal bacteraemia and discordant empiric antibiotic therapy. Currently, early mortality is relatively low and is caused by pneumonia-related factors. It occurs mainly among the elderly and in patients presenting with altered mental status, multilobar pneumonia and septic shock. Pneumococcal bacteraemia and discordant antibiotic therapy, mainly due to lack of coverage against Pseudomonas aeruginosa are also significant risk factors. PMID- 18508821 TI - Budesonide prevents but does not reverse sustained airway hyperresponsiveness in mice. AB - Despite the effectiveness of corticosteroids at resolving airway inflammation, they are only moderately effective at attenuating airway hyperresponsiveness (AHR). The extent to which corticosteroids are able to reverse or inhibit the development of sustained AHR is not known. The present study aimed to determine whether budesonide can resolve and or prevent the development of sustained AHR in mice. Mice were chronically exposed to allergen and treated with budesonide either: 1) briefly during the final weeks of exposure to allergen; 2) prolonged concurrently throughout exposure to allergen; or 3) delayed following final exposure to allergen. AHR was assessed 24 h (brief treatment) or 4 weeks (prolonged concurrent and delayed treatments) following final exposure to allergen. Brief budesonide intervention significantly attenuated the inflammation associated AHR assessed immediately following final exposure to allergen. Similarly, prolonged concurrent budesonide treatment prevented the development of sustained AHR. Delayed budesonide intervention, however, did not resolve sustained AHR. In conclusion, the early introduction and, importantly, the persistence of corticosteroid treatment prevented the development of sustained airway hyperresponsiveness; however, the inability of corticosteroids to reverse established airway dysfunction indicates a limitation in their use for the complete, long-term management of airway hyperresponsiveness. PMID- 18508822 TI - Outdoor swimming pools and the risks of asthma and allergies during adolescence. AB - Exposure to indoor chlorinated swimming pools can be detrimental to the airways of swimmers and increase asthma risks but it is unknown whether these effects concern outdoor pools. The present study examined 847 secondary school adolescents who had attended residential or nonresidential outdoor chlorinated pools at a variable rate. The main outcomes were: ever asthma (physician diagnosed at any time); current asthma (ever asthma under medication and/or with exercise-induced bronchoconstriction); elevated exhaled nitric oxide; and aeroallergen-specific immunoglobulin (Ig)E in serum. The prevalence of ever and current asthma significantly increased with the lifetime number of hours spent in outdoor pools by up to four and eight times, respectively, among adolescents with the highest attendance (>500 h) and a low exposure to indoor pools (<250 h). Odds for asthma were significantly increased among adolescents with total serum IgE >25 kIU x L(-1), on average by 1-2 units for each 100-h increase in pool attendance. Use of residential outdoor pools was also associated with higher risks of elevated exhaled nitric oxide and sensitisation to cat or house dust mite allergens. Outdoor chlorinated swimming pool attendance is associated with higher risks of asthma, airways inflammation and some respiratory allergies. PMID- 18508823 TI - Antimicrobial peptides in lung transplant recipients with bronchiolitis obliterans syndrome. AB - Mechanisms other than classical alloimmunity are implicated in the pathogenesis of bronchiolitis obliterans syndrome (BOS). It was hypothesised that antimicrobial peptides (AMPs), elements of the innate immune response, have a role in BOS pathogenesis. Pulmonary expression of the neutrophil-derived AMPs human cathelicidin (hCAP)-18/LL-37 and alpha-defensins (human neutrophil peptides (HNP) 1-3), and the epithelial cell-derived AMPs human beta-defensin (hBD)-2, elafin and secretory leukoprotease inhibitor (SLPI) were measured in stable lung transplant recipients and those with BOS. The relationship between airway pathogens and AMP levels was examined. Bronchoalveolar lavage (BAL) was performed on 44 lung transplant recipients (30 stable, 14 with BOS). BAL was cultured for pathogens and ELISA for AMPs was performed. The presence of airway pathogens was associated with significantly increased levels of neutrophil-derived and epithelial-derived AMPs. When patients without pathogens in BAL fluid were analysed, eight recipients with BOS had elevated hCAP-18/LL-37 and HNP 1-3 compared with 25 stable recipients. hBD-2 and elafin levels were comparable in BOS and stable recipients, but SLPI levels were reduced in BOS. Bronchiolitis obliterans syndrome is associated with elevated airway human cathelicidin 18/LL 37 and human neutrophil peptides 1-3 from activated neutrophils, even in the absence of pathogens. Together with reduced airway secretory leukoprotease inhibitor this may favour nonalloimmune airway injury with reduced antiprotease defence and increased neutrophil degranulation. PMID- 18508824 TI - Efficacy of a cell phone-based exercise programme for COPD. AB - The application of a supervised endurance exercise training programme in a home setting offering convenience and prolonged effects is a challenge. In total, 48 patients were initially assessed by the incremental shuttle walk test (ISWT), spirometry and the Short Form-12 (SF-12) quality-of-life questionnaire, and then every 4 weeks for 3 months thereafter and again after 1 yr. During the first 3 months, 24 patients in the cell phone group were asked to perform daily endurance walking at 80% of their maximal capacity by following the tempo of music from a program installed on a cell phone. The level of endurance walking at home was readjusted monthly according to the result of ISWT. In the control group, 24 patients received the same protocol and were verbally asked to take daily walking exercise at home. Patients in the cell phone group significantly improved their ISWT distance and duration of endurance walking after 8 weeks. The improvements in ISWT distance, inspiratory capacity and SF-12 scoring at 12 weeks persisted until the end of the study, with less acute exacerbations and hospitalisations. In the present pilot study, the cell phone-based system provides an efficient, home endurance exercise training programme with good compliance and clinical outcomes in patients with moderate-to-severe chronic obstructive pulmonary disease. PMID- 18508825 TI - Workplace-specific challenges as a contribution to the diagnosis of occupational asthma. AB - The diagnosis of occupational asthma can be made by exposing workers to the relevant agent either in a hospital laboratory through specific inhalation challenges (SICs) or in the workplace. As suggested by several authors, workers with negative laboratory SIC can be monitored at the workplace under supervision. The present study aims to assess the frequency of, and identify factors associated with, a positive workplace reaction in workers with negative SIC in the laboratory. The results of workplace challenges were examined in 99 workers who underwent negative SIC between 1994 and 2004. A positive reaction either in the SIC or in the workplace was defined as a sustained fall in forced expiratory volume in one second of > or =20%. In total, 22 (22.2%) workers showed positive responses at the workplace. These subjects more often had increased baseline methacholine responsiveness (90.5 versus 67.6%). They also underwent more days of SIC testing (4.9 versus 3.3 days) and were exposed more often to two or more agents (56 versus 28.4%) and for a longer period of time (363.3 versus 220.4 min) in the laboratory. The present study illustrates the usefulness of workplace monitoring of airway function in the investigation of occupational asthma and identifies factors that are more often associated with a positive reaction. PMID- 18508826 TI - Can CAP guideline adherence improve patient outcome in internal medicine departments? AB - The impact of compliance with Italian guidelines on the outcome of hospitalised community-acquired pneumonia (CAP) in internal medicine departments was evaluated. All Fine class IV or V CAP patients were included in this multicentre, interventional, before-and-after study, composed of three phases: 1) a retrospective phase (RP; 1,443 patients); 2) a guideline implementation phase; and 3) a prospective phase (PP; 1,404 patients). Antibiotic prescription according to the guidelines increased significantly in the PP. The risk of failure at the end of the firstline therapy was significantly lower in the PP versus the RP (odds ratio (OR) 0.83, 95% confidence interval (CI) 0.69-1.00), particularly in Fine class V patients (OR 0.71, 95% CI 0.51-0.98). Analysis of outcome in the overall population (2,847 patients) showed a statistically significant advantage for compliant versus noncompliant therapies in terms of failure rate (OR 0.74, 95% CI 0.60-0.90) and an advantage in terms of mortality (OR 0.77, 95% CI 0.58-1.04). Antipneumococcal cephalosporin monotherapy was associated with a low success rate (68.6%) and the highest mortality (16.2%); levofloxacin alone and the combination of cephalosporin and macrolide resulted in higher success rates (79.1 and 76.7%, respectively) and significantly lower mortalities (9.1 and 5.7%, respectively). Overall, a low compliance with guidelines in the prospective phase (44%) was obtained, indicating the need for future more aggressive and proactive approaches. PMID- 18508827 TI - Haem oxygenase-1 plays a central role in NNK-mediated lung carcinogenesis. AB - The tobacco-specific nitrosamine, 4-(N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1 butanone (NNK), is a potent lung cancer inducer. However, how NNK induces lung cancer is still largely unknown. Haem oxygenase (HO)-1 was evaluated in 30 pairs of lung cancer tumour samples and matched nontumour tissues from patients with a history of cigarette smoking. Expression of HO-1, p21(Cip1/Waf1/Cid1) (p21), B cell lymphoma (Bcl)-2 family members, mitogen-activated protein kinase and nuclear factor (NF)-kappaB was also studied in lung cancer cells treated with NNK. The levels of HO-1 and p21 were significantly increased in lung tumour tissues. There was a positive relationship between these two proteins in the tumour. NNK stimulated lung cell proliferation and elevated the levels of HO-1, p21, inhibitor of apoptosis protein (c-IAP)2 and Bcl-2, but downregulated Bad. These effects of NNK were blocked by zinc protoporphyrin-XII, an HO-1 inhibitor. The NNK-mediated expression of HO-1 was governed by NF-kappaB and extracellular signal-regulated kinase 1/2, since blocking either of these prevented the stimulatory effect of NNK on HO-1, as well as molecules downstream of HO-1, such as p21, c-IAP2, Bcl-2 and Bad. In conclusion, haem oxygenase-1 plays a central role in NNK-mediated cell proliferation by promoting the expression of p21(Cip1/Waf1/Cid1), inhibitor of apoptosis protein 2 and B-cell lymphoma-2 but inhibiting the activity of Bad. Nuclear factor-kappaB and extracellular signal regulated kinase 1/2 function upstream of haem oxygenase-1. Therefore, haem oxygenase-1 is likely to be a potential target in the treatment of smoking related lung cancer. PMID- 18508828 TI - Treatment with inhaled steroids in patients with symptoms suggestive of asthma but with normal lung function. AB - A total of 144 patients with lower airway symptoms suggestive of asthma, but who did not fulfil the functional criteria of asthma, were included in a randomised, double-blind, placebo-controlled 8-week "proof-of-concept" study with mometasone furoate (MF), 400 microg once daily. The primary efficacy variable was the mean change from baseline in six morning and evening weekly symptom scores: cough, sputum production, wheeze, shortness of breath, chest tightness and exercise induced cough/wheeze. Total symptom scores were calculated after treatment for 4 and 8 weeks. Compared with placebo, MF improved total morning symptom score at 8 weeks. Changes in total evening symptom scores did not differ between treatments. MF improved all individual symptom scores more than placebo, although the differences in changes between treatments were not always statistically significant. Morning and evening peak expiratory flow rates increased with MF compared with placebo. MF reduced eosinophils and the levels of eosinophilic cationic protein in induced sputum. The results show that symptoms suggestive of asthma exist in patients without significant beta(2)-agonist reversibility or diurnal variability in peak flow. Once-daily MF may benefit some of these patients and a short course with inhaled corticosteroids may be tried. Responders should be better identified in further studies. PMID- 18508829 TI - Adrenal suppression in bronchiectasis and the impact of inhaled corticosteroids. AB - The present study identified three patients with bronchiectasis receiving inhaled corticosteroids (ICSs) who had symptomatic adrenal suppression secondary to ICS. The prevalence of adrenal suppression is unknown in bronchiectasis. The frequency of adrenal suppression and the impact of ICS use in bronchiectasis patients were examined. In total, 50 outpatients (33 receiving ICSs) underwent a short Synacthen test and completed a St George's Respiratory Questionnaire (SGRQ). Symptoms of adrenal suppression, steroid use and lung function were compared between subjects who were suppressed and those who were not. Adrenal suppression was evident in 23.5% of subjects who did not receive ICSs and 48.5% of those who did. Basal cortisol and the increments by which cortisol increased 30 min after Synacthen were lower in suppressed than in nonsuppressed subjects. The incremental cortisol rise was negatively correlated with SGRQ impacts and total score, suggesting a worse quality of life in those who had an impaired adrenal response. The greatest frequency of generalised symptoms was seen in the suppressed group. A significant proportion of subjects with bronchiectasis have evidence of adrenal suppression, and this is increased when inhaled corticosteroids are also used. Impairment of the cortisol response to stimulation is associated with poorer health status. PMID- 18508830 TI - Angiotensinogen gene G-6A polymorphism influences idiopathic pulmonary fibrosis disease progression. AB - Angiotensin II is a growth factor that plays a key role in the physiopathology of idiopathic pulmonary fibrosis (IPF). A nucleotide substitution of an adenine instead of a guanine (G-6A) in the proximal promoter region of angiotensinogen (AGT), the precursor of angiotensin II, has been associated with an increased gene transcription rate. In order to investigate whether the G-6A polymorphism of the AGT gene is associated with IPF development, severity and progression, the present study utilised a case-control study design and genotyped G-6A in 219 patients with IPF and 224 control subjects. The distribution of G-6A genotypes and alleles did not significantly differ between cases and controls. The G-6A polymorphism of the AGT gene was not associated with disease severity at diagnosis. The presence of the A allele was strongly associated with increased alveolar arterial oxygen tension difference during follow-up, after controlling for the confounding factors. Higher alveolar arterial oxygen tension changes over time were observed in patients with the AA genotype (0.37+/-0.7 mmHg (0.049+/ 0.093 kPa) per month) compared to GA genotype (0.12+/-1 mmHg (0.016+/-0.133 kPa) per month) and GG genotype (0.2+/-0.6 mmHg (0.027+/-0.080 kPa) per month). G-6A polymorphism of the angiotensinogen gene is associated with idiopathic pulmonary fibrosis progression but not with disease predisposition. This polymorphism could have a predictive significance in idiopathic pulmonary fibrosis patients. PMID- 18508831 TI - Effects of glucocorticoids in ventilated piglets with severe pneumonia. AB - There is clinical evidence suggesting that glucocorticoids may be useful in severe pneumonia, but the pathogenic mechanisms explaining these beneficial effects are unknown. The aim of the present study was to determine the effects of adding glucocorticoids to antibiotic treatment in an experimental model of severe pneumonia. In total, 15 Lagerwhite-Landrace piglets were ventilated for 96 h. After intubation, a 75 mL solution containing Pseudomonas aeruginosa (10(6) cfu x mL(-1)) was bronchoscopically inoculated. The animals were randomised into three groups 12 h after inoculation: 1) untreated; 2) treated with ciprofloxacin; and 3) treated with ciprofloxacin plus methylprednisolone. Physiological and laboratory parameters were monitored throughout the study. Pro-inflammatory cytokines were measured in serum and bronchoalveolar lavage (BAL). Histopathology of the lungs and cultures from blood, BAL and lungs were performed. At the end of the study, piglets receiving the antibiotic plus glucocorticoids showed: 1) a decrease in the concentration of interleukin-6 in BAL; and 2) a decrease in the global bacterial burden both in BAL and lung tissue. In conclusion, in this experimental model of pneumonia, the association of glucocorticoids with antibiotics attenuates local inflammatory response and decreases bacterial burden in the lung. PMID- 18508832 TI - CPAP decreases plasma levels of soluble tumour necrosis factor-alpha receptor 1 in obstructive sleep apnoea. AB - There is increasing evidence that inflammation plays an important role in the development of cardiovascular complications in patients with obstructive sleep apnoea (OSA). No previous works have studied levels of soluble tumour necrosis factor-alpha receptor (sTNFR)-1 in patients with OSA. The aims of the present study were to examine serum levels of sTNFR-1 and the effect of nasal continuous positive airway pressure (CPAP) in patients with OSA. A prospective, randomised, placebo-controlled crossover study was performed. In total, 30 consecutive newly diagnosed OSA patients (apnoea/hypopnoea index 43.8+/-27.0 events x h(-1)) and 15 healthy obese patients were selected. Urinary levels of norepinephrine and epinephrine, as well as plasma sTNFR-1, tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and leukotriene (LT)B(4) levels were obtained at baseline and after 3 months of CPAP or sham CPAP. Nocturnal urinary levels of norepinephrine, epinephrine and sTNFR-1 (1,053+/-269 versus 820+/-166 pg x mL(-1)) were significantly higher in OSA patients. There were no significant differences in plasma levels of IL-6, LTB(4), or TNF-alpha between the two study groups. There were no significant differences in blood pressure, urinary catecholamine levels, or plasma IL-6, LTB(4) and TNF-alpha levels after both treatment modalities. However, after 3 months of effective CPAP usage, sTNFR-1 levels were significantly reduced (1,053+/-269 versus 899+/-254 pg x mL(-1)). Obstructive sleep apnoea patients have higher levels of soluble tumour necrosis factor-alpha receptor 1 than individuals without OSA; soluble tumour necrosis factor-alpha receptor 1 levels are lowered by continuous positive airway pressure therapy. These findings further corroborate a potential role of inflammation in the natural history of obstructive sleep apnoea. PMID- 18508833 TI - Usefulness of consecutive C-reactive protein measurements in follow-up of severe community-acquired pneumonia. AB - Despite the introduction of new inflammatory markers, C-reactive protein (CRP) remains commonly used in patients hospitalised with severe infections. However, evidence on the usefulness of consecutive CRP measurements is still unclear. The clinical relevance of consecutive CRP measurements was studied in follow-up of antibiotic treatment in patients with severe community-acquired pneumonia (CAP). In a prospective multicentre trial, CRP levels were measured on admission, and on days 3 and 7. Patients were followed clinically for 28 days. Aetiology could be determined in 137 (47.4%) out of the 289 patients included. In 122 (38.8%) patients, initial antibiotic therapy was appropriate. A decline of <60% in CRP levels in 3 days and a decline of <90% in CRP levels in 7 days were both associated with an increased risk of having received inappropriate empiric antibiotic treatment (day 0-3, odds ratio (OR) 6.98, 95% confidence interval (CI) 1.56-31.33 and day 0-7, OR 3.74, 95% CI 1.12-13.77). In conclusion, consecutive C reactive protein measurements are useful in the first week in follow-up of antibiotic treatment for severe community-acquired pneumonia when taking the causative microorganism and use of steroids into account. A delayed normalisation of C-reactive protein levels is associated with a higher risk of having received inappropriate antibiotic treatment. PMID- 18508834 TI - Symptomatic mucocele after esophageal exclusion. AB - Surgical exclusion of the thoracic esophagus can result in the accumulation of secretions and dilatation of the esophageal remnant, a clinical picture known as esophageal mucocele. Although it is usually asymptomatic, if it increases in size it can produce a variety of compressive symptoms such as coughing, chest pain and respiratory distress. We present two cases of symptomatic mucocele after esophageal exclusion treated successfully with surgical resection. We believe that surgical resection should be considered for symptomatic patients, and that esophageal bypass surgery should be used with caution and indicated mostly in patients with a limited life span or with contraindications for esophagectomy. PMID- 18508835 TI - The influence of low dialysate sodium and glucose concentration on volume distributions in body compartments after haemodialysis: a bioimpedance analysis study. AB - BACKGROUND: Despite the developments in haemodialysis, there are still some difficulties in maintaining the haemodynamic stability. Bioimpedance analysis (BIA) has been proposed for the estimation of dry weight in haemodialysis patients. We aimed to investigate the effects of dialysate sodium and glucose contents on volume distribution in body compartments after haemodialysis by using BIA, a sensitive and reliable method. METHODS: Seventeen chronic haemodialysis patients [11 males, 6 females, mean age: 36.9 (18-64) years] were included in the study. Patients were evaluated in three periods. The patients (period 1-P1) underwent haemodialysis with dialysate of 200 mg/dL glucose and 140 mmol/dL sodium for 4.5 h in the middle session of the first week. At the beginning and the end of the session, haematocrit, vital parameters (blood pressure, pulse), ultrafiltrated volume, plasma osmolarity and plasma renin activity were recorded. Also multi-frequency bioelectric impedance analyses (Bodystat Quadscan 4000) were applied to all patients at 5, 50, 100 and 200 kHz, including the impedance index (Z200/Z5). In the second midweek session the same procedure was repeated with same glucose concentration and 135 mmol/dL sodium including dialysate (period 2 P2), and in the third week, it was performed with a dialysate that included 140 mmol/dL sodium and no glucose (period 3-P3). RESULTS: The change of the ratio of the intracellular volume to total body weight (ICV/TBW) at the beginning and the end of the session was same in all periods. However, there were significant differences in the change (after/before session) ratio for the extracellular volume/total body weight (ECV/TBW) in P2 compared to other periods (P values for P1-P2: <0.001 and P2-P3: 0.007). Likewise, the same was observed in the changes of impedance (P values for P1-P2: 0.08, P1-P3: 0.44 and P2-P3: 0.063). There was a significant increase of hypotensive events in P2 against the other periods (P = 0.001). CONCLUSION: Decreasing dialysate sodium concentration results in important haemodynamic changes but the lack of glucose in dialysate does not result in any changes in haemodynamic and inflammatory parameters. The changes in bioimpedance parameters are parallel to haemodynamic changes in the haemodialysis patients. PMID- 18508836 TI - Cyclic appearance of left ventricular outflow tract dynamic obstruction during mechanical ventilation: evidence for a preload dependent phenomenon. AB - The cyclic appearance of dynamic left ventricular outflow tract obstruction during mechanical ventilation, according to the phasic changes in preload, is described in this article. Hemodialysis-induced fluid removal resulted in preload dependence as evidenced by the pulse pressure variation in a 56-year-old critically ill patient. The clinical picture was suggestive of myocardial failure. Transthoracic echocardiography disclosed dynamic left ventricular outflow tract obstruction associated with systolic anterior motion of the mitral valve. Progressive fluid restitution resulted in a parallel decrease in both the degree of dynamic obstruction and pulse pressure variation. During fluid loading, dynamic obstruction disappeared at first during the inspiratory phase of intermittent positive pressure ventilation corresponding to the phasic increase in left ventricular preload. Further fluid loading resulted in the disappearance of dynamic obstruction during both inspiratory and expiratory phase of intermittent positive pressure ventilation. This is the first reported case clearly relating left ventricular outflow tract dynamic obstruction to preload dependence during mechanical ventilation in a critically ill patient without predisposing anatomical factor. PMID- 18508837 TI - Acute respiratory distress syndrome from chlorine inhalation during a swimming pool accident: a case report and review of the literature. AB - Chlorine inhalation can result in significant morbidity and mortality. The most common clinical ramification is mucosal irritation. Rarely, depending upon the degree of exposure, patients can develop acute respiratory distress syndrome. Management is usually supportive with an unproven role for inhaled or systemic corticosteroids. A case of a young woman who developed respiratory failure secondary to acute respiratory distress syndrome from accidental exposure to chlorine fumes at a community swimming pool is described. The patient suffered a prolonged hospitalization with the need for mechanical ventilation. Despite limited data to support the decision, the patient was started on treatment with corticosteroids. She recovered completely from her illness and was discharged home without supplemental oxygen. A concise discussion of chlorine inhalation injury and a literature review on the utility of inhaled and/or systemic corticosteroids for this clinical entity is presented. PMID- 18508838 TI - Bispectral index monitoring documents burst suppression during pentobarbital coma. AB - During pentobarbital coma, electroencephalographic monitoring is used to document burst suppression (3-5 episodes of electrical activity/min). The current study evaluates the association of the bispectral index number and suppression ratio with a burst suppression pattern on electroencephalograph. The records of 7 patients (aged 2.9-14 years) who received pentobarbital for elevated intracranial pressure were retrospectively reviewed. The bispectral index number was 7 +/- 5, 14 +/- 3, and 37 +/- 12, whereas the suppression ratio was 93 +/- 7%, 75 +/- 6%, and 29 +/- 18% when the electroencephalograph showed or= 6 bursts/min, respectively. The sensitivity and specificity of a bispectral index value of 10 to 20 were 96% and 92%, respectively, whereas the sensitivity and specificity of a suppression ratio of 65% to 85% were 89% and 88%, respectively, in demonstrating the presence of 3 to 5 bursts/min. Bispectral index monitoring may be easier to perform and may require less technical expertise to interpret. PMID- 18508839 TI - A microcosting study of intensive care unit stay in the Netherlands. AB - The primary objective of this study was to estimate the actual daily costs of intensive care unit stay using a microcosting methodology. As a secondary objective, the degree of association between daily intensive care unit costs and some patient characteristics was examined. This multicenter, retrospective cost analysis was conducted in the medical-surgical adult intensive care units of 1 university and 2 general hospitals in the Netherlands for 2006, from a hospital perspective. A total of 576 adult patients were included, consuming a total of 2868 nursing days. The mean total costs per intensive care unit day were 1911, with labour (33%) and indirect costs (33%) as the most important cost drivers. An ordinary least squares analysis including age, Nine Equivalent of Nursing Manpower Use score/Therapeutic Intervention Scoring System score, mechanical ventilation, blood products, and renal replacement therapy was able to predict 50% of the daily intensive care unit costs. PMID- 18508840 TI - Del(5q) is associated with clinical and histological parameters in small cell neuroendocrine lung carcinoma. AB - To elucidate the relationship between del(5q) and the clinical and histological features of small cell neuroendocrine lung carcinoma, 33 tissue samples from patients with this tumor were evaluated. By using fluorescence in situ hybridization, del(5q) was identified in almost 50% of cases (15/33, 45%). Clinically, patients with tumors showing del(5q) were older (mean age = 71 years) with a correspondingly greater pack-year smoking history (mean = 61) than patients with tumors (mean age = 59 years, mean pack-years = 44) without del(5q). Histologically, tumors with del(5q) had a greater frequency of spindle cell morphology (11/14 [79%] vs 6/16 [38%], P < .025) than those without del(5q). This is the first study to find an association between del(5q) and tumor histology in small cell neuroendocrine lung carcinoma. PMID- 18508841 TI - A novel method of obtaining prostate tissue for gene expression profiling. AB - Gene expression profiling by DNA microarray analysis is a technique with great promise in cancer biology. The multifocality and heterogeneity of many prostate cancers makes the collection of adequate biological samples for such profiling particularly challenging. Current methods, such as laser capture microdissection, are not widely available and can have significant limitations. In this article, a novel method of prostatic sampling, which does not affect the histopathological assessment of the surgical specimen and provides adequate RNA yield for microarray analysis is described. This method is simple, inexpensive, easily reproducible, and has been validated as having >95% sensitivity and 99% specificity for histological prediction of tissue obtained. This method can be adopted by other investigators to perform DNA microarray analysis on prostate tumors. PMID- 18508842 TI - Primary carcinosarcoma of the spleen: case report of a rare tumor and review of the literature. AB - Carcinosarcoma of extragenital organs is rare. In this article, a case of primary carcinosarcoma of the spleen, which presented as painful splenomegaly is reported. To the best of our knowledge, this is the second reported case of primary splenic carcinosarcoma in English literature. The pathogenesis of these tumors is incompletely understood. No specific treatment guidelines exist for these aggressive tumors, surgery being the mainstay of treatment. The prognosis remains poor regardless of the adjuvant therapy used. PMID- 18508843 TI - Small-cell neuroendocrine carcinoma of the nasopharynx: report of a rare case lacking association with Epstein-Barr virus. AB - Only 2 cases of nasopharyngeal small cell carcinoma have been reported. Another case of this rare type of nasopharyngeal tumor confirmed by immunopositivity for CD56, synaptophysin, and chromogranin A is described in this study. The tumor was also negative for cytokeratin (CK) 20 and thyroid transcription factor-1. Clinical investigation did not find any primary tumor in other anatomic sites. Therefore, this case was established as a primary nasopharyngeal small cell carcinoma. Epstein-Barr virus was shown to be absent by Epstein-Barr virus encoded RNA in situ hybridization study in this case, which further distinguished it from conventional nasopharyngeal carcinoma. Clinical features of this case and 2 previously reported cases are compared. They do not differ from conventional nasopharyngeal carcinoma in clinical presentation, but the prognosis seems to be worse. More cases are needed to confirm the observation and for understanding the clinical behavior of this rare type of nasopharyngeal tumor. PMID- 18508844 TI - Malakoplakia of the spleen: a case report. AB - Malakoplakia is an uncommon chronic inflammatory disorder, which is characterized by the presence of histiocytes containing concentric concretions known as Michaelis-Gutmann bodies in a background of mixed inflammation. The urinary tract is the most commonly involved site. However, malakoplakia can be found in a wide range of other organs throughout the body. Its occurrence has been attributed to a defect in the bactericidal capacity of phagocytic cells, and it is usually seen in patients with some degree of immunologic compromise. A case of malakoplakia of the spleen in a patient with Crohn's disease is reported in this article. PMID- 18508845 TI - Plasmablastic lymphoma affecting the lung and bone marrow with CD10 expression and t(8;14)(q24;q32) translocation. AB - Plasmablastic lymphoma is a rare variant of a diffuse, large B-cell lymphoma, which typically presents in the oral cavity in immunocompromised patients. In HIV positive patients, this tumor has a tendency to manifest in extramedullary sites. In this report, we document a rare instance in which this neoplasm besides affecting the bone marrow also involved the lung. In addition, the lymphoma in our case disclosed CD10 positivity on immunohistochemistry and t(8;14)(q24;q34) translocation on cytogenetic analysis, mimicking a Burkitt/atypical Burkitt lymphoma. The problems in diagnosis are discussed. PMID- 18508846 TI - Histopathological diagnosis of intestinal spirochetosis in a nonimmunocompromised patient. PMID- 18508847 TI - Statins and heart failure. AB - Retrospective studies show that statin use prevents congestive heart failure and decreases mortality in patients with congestive heart failure; however, only one prospective study is available. In the Controlled Rosuvastatin Multinational Trial in Heart Failure study, rosuvastatin (10 mg/d) was compared with placebo in patients with advanced coronary heart disease-related congestive heart failure. There was no significant difference between rosuvastatin and placebo except for hospitalization rates. Statins must be given to all patients with coronary heart disease as early as possible because they may not work in patients with advanced congestive heart failure. PMID- 18508848 TI - Predictors for peripheral and carotid revascularization in a population-based cohort with type 2 diabetes. AB - The study was aimed to define the predictors for peripheral and carotid revascularization in type 2 diabetic population. In all, 279 patients with type 2 diabetes and peripheral arterial disease were enrolled in a cohort longitudinal study. Study population was followed up for 24 months for the need and performance of peripheral or carotid revascularization. Logistic regression analysis was conducted to identify variables predictive of revascularization, when lipid plasma levels, glycemia, arterial hypertension, blood pressures, ankle brachial index, intima-media thickness, body mass index, waist circumference, and hip distances were put in a model. Total cholesterol and maximal value of carotid intima-media thickness were presented as factors that independently influenced the performed peripheral revascularization. Waist circumference is defined as independent factor associated with carotid endarterectomy. Measurement of carotid intima-media thickness, waist circumference, and plasma lipid levels in type 2 diabetes with manifested peripheral and carotid arterial disease should be recommended in a manner of proper risk stratification of this population. PMID- 18508849 TI - Percutaneous revascularization of grafts versus native coronary arteries in postcoronary artery bypass graft patients. AB - In patients with previous coronary artery bypass graft surgery, it is unknown whether better results may be obtained with percutaneous interventions of grafts versus native arteries. The clinical outcomes in 84 patients undergoing percutaneous interventions of either grafts (n = 31) or native arteries (n = 53) were compared. Procedural success rate was 95.3% (96.8% in the graft group vs 94.4% in the native group, P = .3). Mean follow-up was 19 +/- 7 months. The incidence of major adverse events was 14.2% (12.9% vs 15.1% in the graft and native groups, respectively; P = .8), mortality rate was 3.5% (6.4% vs 1.8% in the graft and native groups, respectively; P = .3), and target-lesion revascularization was performed in 4.7% (6.4% vs 3.7% in the graft and native groups, respectively, P = .6). In conclusion, both graft or native percutaneous interventions were similar for immediate and midterm clinical outcomes. The relatively low risk need for target-lesion revascularization obtained with both strategies is encouraging. PMID- 18508850 TI - Physical activity and cardiovascular disease prevention: current recommendations. AB - Epidemiologic studies support an inverse and robust relationship between physical activity (PA) and mortality risk, even after adjusting for potential confounding factors. The authors have reported 13% lower mortality risk for every 1-MET (metabolic equivalent) increase in exercise capacity. For those with an exercise capacity >7 METs, the mortality risk was approximately 50% to 70% lower when compared with those achieving <5 METs. The risk reduction is, at least in part, attributed to the favorable effect of PA on the cardiovascular risk factors. Increased PA lowers blood pressure in hypertensive individuals, increases high density lipoprotein cholesterol in a dose-response fashion, and reduces the incidence of diabetes. The health benefits of PA can be realized by engaging in moderate-intensity physical activity (brisk walk) for at least 30 minutes per day, 5 days per week or vigorous activity (jogging) for 20 or more minutes, 3 days per week. Combinations of the 2 types of activity can also be performed. PMID- 18508851 TI - One-year follow-up after implantation of the EverFlex nitinol stent in long lesions of the superficial femoral artery. AB - For treatment of complex superficial femoral artery lesions, suitable stent devices are required. The present study details the first long-term results with the long EverFlex nitinol stent. Forty-one EverFlex stents were implanted in 32 patients with either superficial femoral artery occlusions (n = 20) or stenoses (n = 12); mean lesion length was 13.3 cm (range = 8.0-39.0). Patients presented with clinical stage Fontaine IIb (n = 27), III (n = 4), or IV (n = 1). Stent lengths were 10 cm (n = 18), 12 cm (n = 6), or 15 cm (n = 17). Follow-up after 2 months indicated primary and secondary patency rates of 96.9% (n = 31) and 100% (n = 32); ABI improved from 0.57 to 0.91 (P < .001). After 1 year, revascularization was performed in 6 patients (18.8%). Primary and secondary patency rates were 81.3% (n = 26) and 93.8% (n = 30). In this first long-term evaluation, the long EverFlex nitinol stent achieves excellent clinical results after implantation into complex superficial femoral artery lesions. PMID- 18508852 TI - Comparison of quantitative coronary angiography with intracoronary ultrasound. Can quantitative coronary angiography accurately estimate the severity of a luminal stenosis? AB - In this study we investigated the accuracy of monoplane and biplane quantitative coronary angiography in estimating the luminal dimensions, using intracoronary ultrasound as gold standard. Biplane angiography and intracoronary ultrasound were performed in 24 arterial segments. The end-diastolic intracoronary ultrasound frames were manually selected and segmented. In 2 end-diastolic X ray projections, quantitative coronary angiography was performed and a novel methodology was applied to register the segmented frames onto the processed angiographic images. The luminal areas determined by quantitative coronary angiography in 1 (monoplane) and 2 projections (mean) were compared with those determined by intracoronary ultrasound. The obtained correlation coefficients for the monoplane and mean estimations were 0.69 +/- 0.12 and 0.77 +/- 0.08, respectively. It would appear that by increasing the angle between the biplane projections, the correlation between intracoronary ultrasound and mean estimations improves. Our results provide evidence that orthogonal biplane angiography is more reliable and should be preferred to assess luminal dimensions. PMID- 18508853 TI - Avoiding and managing bleeding complications in patients with non-ST-segment elevation acute coronary syndromes. AB - Antithrombotic therapy coupled with early use of cardiac catheterization and revascularization have decreased morbidity and mortality rates in patients who have acute ischemic heart disease but who carry a risk for bleeding. Bleeding complications in patients with acute coronary syndromes are associated with worse clinical outcomes, including recurrent ischemic events and death. Determining the appropriate balance between preventing ischemic events and causing bleeding in patients with acute coronary syndromes present a challenging problem for clinicians. Antithrombotics studied in recent clinical trials that have focused on bleeding reduction include bivalirudin and fondaparinux. In this review, the incidence, predictors, and clinical outcomes associated with bleeding are discussed. Furthermore, the association between antithrombotic agents and bleeding and propose strategies to prevent bleeding complications are also discussed. PMID- 18508854 TI - Overexpression of endoplasmic reticulum-resident chaperone attenuates cardiomyocyte death induced by proteasome inhibition. AB - AIMS: Proteasome inhibitors are a novel class of anticancer agents that induce tumour cell death via endoplasmic reticulum (ER) stress. Since ER stress is involved in the development of heart failure, we investigated the role of ER initiated cardiomyocyte death by proteasome inhibition. METHODS AND RESULTS: Rat neonatal cardiomyocytes were used in this study. Proteasome activity was assayed using proteasome peptidase substrates. Cell viability and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenol tetrazolium bromide and flow cytometry, respectively. Western blot analysis, real-time polymerase chain reaction (PCR) and reverse transcriptional PCR were used to detect the expression of protein and messenger ribonucleic acid (RNA). The location of overexpressed glucose-regulated protein (GRP) 78 was observed by confocal fluorescence microscopy. Proteasome inhibition induced cardiomyocyte death and activated ER stress-induced transcriptional factor ATF6, but not XBP1 (X-box binding protein 1), without up-regulating ER chaperones. ER-initiated apoptosis signalling, including cytosine-cytosine-adenine-adenine-thymine enhancer-binding protein (C/EBP) homologous protein (CHOP), c-Jun-N-terminal kinase (JNK), and caspase-12, was activated by proteasome inhibition. Short interference RNA targeting CHOP, but not the blockage of caspase-12 or JNK pathway, attenuated cardiomyocyte death. Overexpression of GRP78 suppressed both CHOP expression and cardiomyocyte death by proteasome inhibition. CONCLUSION: These findings demonstrate that proteasome inhibition induces ER-initiated cardiomyocyte death via CHOP-dependent pathways without compensatory up-regulation of ER chaperones. Supplement and/or pharmacological induction of GRP78 can attenuate cardiac damage by proteasome inhibition. PMID- 18508855 TI - Towards the second generation of skeletal myoblasts? PMID- 18508856 TI - InteroPORC: automated inference of highly conserved protein interaction networks. AB - MOTIVATION: Protein-protein interaction networks provide insights into the relationships between the proteins of an organism thereby contributing to a better understanding of cellular processes. Nevertheless, large-scale interaction networks are available for only a few model organisms. Thus, interologs are useful for a systematic transfer of protein interaction networks between organisms. However, no standard tool is available so far for that purpose. RESULTS: In this study, we present an automated prediction tool developed for all sequenced genomes available in Integr8. We also have developed a second method to predict protein-protein interactions in the widely used cyanobacterium Synechocystis. Using these methods, we have constructed a new network of 8783 inferred interactions for Synechocystis. AVAILABILITY: InteroPORC is open-source, downloadable and usable through a web interface at http://biodev.extra.cea.fr/interoporc/. PMID- 18508857 TI - Multistep, sequential control of the trafficking and function of the multiple sulfatase deficiency gene product, SUMF1 by PDI, ERGIC-53 and ERp44. AB - Sulfatase modifying factor 1 (SUMF1) encodes for the formylglicine generating enzyme, which activates sulfatases by modifying a key cysteine residue within their catalytic domains. SUMF1 is mutated in patients affected by multiple sulfatase deficiency, a rare recessive disorder in which all sulfatase activities are impaired. Despite the absence of canonical retention/retrieval signals, SUMF1 is largely retained in the endoplasmic reticulum (ER), where it exerts its enzymatic activity on nascent sulfatases. Part of SUMF1 is secreted and paracrinally taken up by distant cells. Here we show that SUMF1 interacts with protein disulfide isomerase (PDI) and ERp44, two thioredoxin family members residing in the early secretory pathway, and with ERGIC-53, a lectin that shuttles between the ER and the Golgi. Functional assays reveal that these interactions are crucial for controlling SUMF1 traffic and function. PDI couples SUMF1 retention and activation in the ER. ERGIC-53 and ERp44 act downstream, favoring SUMF1 export from and retrieval to the ER, respectively. Silencing ERGIC 53 causes proteasomal degradation of SUMF1, while down-regulating ERp44 promotes its secretion. When over-expressed, each of three interactors favors intracellular accumulation. Our results reveal a multistep control of SUMF1 trafficking, with sequential interactions dynamically determining ER localization, activity and secretion. PMID- 18508858 TI - The perspective and role of the medical oncologist in cancer prevention: a position paper by the European Society for Medical Oncology. PMID- 18508859 TI - EGFR regulation by microRNA in lung cancer: a rose by any other name ... is an increasingly complicated rose. PMID- 18508860 TI - Localised axial progenitor cell populations in the avian tail bud are not committed to a posterior Hox identity. AB - The outgrowth of the vertebrate tail is thought to involve the proliferation of regionalised stem/progenitor cell populations formed during gastrulation. To follow these populations over extended periods, we used cells from GFP-positive transgenic chick embryos as a source for donor tissue in grafting experiments. We determined that resident progenitor cell populations are localised in the chicken tail bud. One population, which is located in the chordoneural hinge (CNH), contributes descendants to the paraxial mesoderm, notochord and neural tube, and is serially transplantable between embryos. A second population of mesodermal progenitor cells is located in a separate dorsoposterior region of the tail bud, and a corresponding population is present in the mouse tail bud. Using heterotopic transplantations, we show that the fate of CNH cells depends on their environment within the tail bud. Furthermore, we show that the anteroposterior identity of tail bud progenitor cells can be reset by heterochronic transplantation to the node region of gastrula-stage chicken embryos. PMID- 18508861 TI - Control of cell flattening and junctional remodeling during squamous epithelial morphogenesis in Drosophila. AB - Diverse types of epithelial morphogenesis drive development. Similar cytoskeletal and cell adhesion machinery orchestrate these changes, but it is unclear how distinct tissue types are produced. Thus, it is important to define and compare different types of morphogenesis. We investigated cell flattening and elongation in the amnioserosa, a squamous epithelium formed at Drosophila gastrulation. Amnioserosa cells are initially columnar. Remarkably, they flatten and elongate autonomously by perpendicularly rotating the microtubule cytoskeleton--we call this 'rotary cell elongation'. Apical microtubule protrusion appears to initiate the rotation and microtubule inhibition perturbs the process. F-actin restrains and helps orient the microtubule protrusions. As amnioserosa cells elongate, they maintain their original cell-cell contacts and develop planar polarity. Myosin II localizes to anterior-posterior contacts, while the polarity protein Bazooka (PAR 3) localizes to dorsoventral contacts. Genetic analysis revealed that Myosin II and Bazooka cooperate to properly position adherens junctions. These results identify a specific cellular mechanism of squamous tissue morphogenesis and molecular interactions involved. PMID- 18508862 TI - mls-2 and vab-3 Control glia development, hlh-17/Olig expression and glia dependent neurite extension in C. elegans. AB - Glia are essential components of nervous systems. However, genetic programs promoting glia development and regulating glia-neuron interactions have not been extensively explored. Here we describe transcriptional programs required for development and function of the C. elegans cephalic sheath (CEPsh) glia. We demonstrate ventral- and dorsal-restricted roles for the mls-2/Nkx/Hmx and vab 3/Pax6/Pax7 genes, respectively, in CEPsh glia differentiation and expression of the genes hlh-17/Olig and ptr-10/Patched-related. Using mls-2 and vab-3 mutants, as well as CEPsh glia-ablated animals, we show that CEPsh glia are important for sensory dendrite extension, axon guidance/branching within the nerve ring, and nerve ring assembly. We demonstrate that UNC-6/Netrin, expressed in ventral CEPsh glia, mediates glia-dependent axon guidance. Our results suggest possible similarities between CEPsh glia development and oligodendrocyte development in vertebrates, and demonstrate that C. elegans provides a unique environment for studying glial functions in vivo. PMID- 18508864 TI - Timing of Wingless signalling distinguishes maxillary and antennal identities in Drosophila melanogaster. AB - The Drosophila adult head mostly derives from the composite eye-antenna imaginal disc. The antennal disc gives rise to two adult olfactory organs: the antennae and maxillary palps. Here, we have analysed the regional specification of the maxillary palp within the antennal disc. We found that a maxillary field, defined by expression of the Hox gene Deformed, is established at about the same time as the eye and antennal fields during the L2 larval stage. The genetic program leading to maxillary regionalisation and identity is very similar to the antennal one, but is distinguished primarily by delayed prepupal expression of the ventral morphogen Wingless (Wg). We find that precociously expressing Wg in the larval maxillary field suffices to transform it towards antennal identity, whereas overexpressing Wg later in prepupae does not. These results thus indicate that temporal regulation of Wg is decisive to distinguishing maxillary and antennal organs. Wg normally acts upstream of the antennal selector spineless (ss) in maxillary development. However, mis-expression of Ss can prematurely activate wg via a positive-feedback loop leading to a maxillary-to-antenna transformation. We characterised: (1) the action of Wg through ss selector function in distinguishing maxillary from antenna; and (2) its direct contribution to identity choice. PMID- 18508863 TI - Embryonic requirements for ErbB signaling in neural crest development and adult pigment pattern formation. AB - Vertebrate pigment cells are derived from neural crest cells and are a useful system for studying neural crest-derived traits during post-embryonic development. In zebrafish, neural crest-derived melanophores differentiate during embryogenesis to produce stripes in the early larva. Dramatic changes to the pigment pattern occur subsequently during the larva-to-adult transformation, or metamorphosis. At this time, embryonic melanophores are replaced by newly differentiating metamorphic melanophores that form the adult stripes. Mutants with normal embryonic/early larval pigment patterns but defective adult patterns identify factors required uniquely to establish, maintain or recruit the latent precursors to metamorphic melanophores. We show that one such mutant, picasso, lacks most metamorphic melanophores and results from mutations in the ErbB gene erbb3b, which encodes an EGFR-like receptor tyrosine kinase. To identify critical periods for ErbB activities, we treated fish with pharmacological ErbB inhibitors and also knocked down erbb3b by morpholino injection. These analyses reveal an embryonic critical period for ErbB signaling in promoting later pigment pattern metamorphosis, despite the normal patterning of embryonic/early larval melanophores. We further demonstrate a peak requirement during neural crest migration that correlates with early defects in neural crest pathfinding and peripheral ganglion formation. Finally, we show that erbb3b activities are both autonomous and non-autonomous to the metamorphic melanophore lineage. These data identify a very early, embryonic, requirement for erbb3b in the development of much later metamorphic melanophores, and suggest complex modes by which ErbB signals promote adult pigment pattern development. PMID- 18508865 TI - Maintaining standards: promoting equality. PMID- 18508866 TI - Health, work and the general practitioner. PMID- 18508867 TI - Why I became an occupational physician ... PMID- 18508868 TI - John Cooke Bourne, Working Shaft Kilsby Tunnel, 8 July 1837. Wash drawing, 19.5 x 20.4 cm. National Railway Museum, York. PMID- 18508869 TI - Diabetes and depression--is there a link to the HPA axis? PMID- 18508871 TI - Analysis of the factors influencing dysphagia severity upon diagnosis of head and neck cancer. AB - Our aim was to assess the influence of age, co-morbidity factors and tumour characteristics on dysphagia severity in the diagnosis of head and neck cancer. Modified barium swallow (MBS) examinations were performed in patients at diagnosis of head and neck cancer. Dysphagia was graded on a scale of 1 to 7 of increasing severity. Between 2000 and 2006, 236 patients with dysphagia underwent MBS at diagnosis of their head and neck cancer. 82 patients were scored as Grade 1, 88 as Grade 2, 29 as Grade 3, 15 as Grade 4, 9 as Grade 5, 5 as Grade 6, and 8 as Grade 7. Grade 3-7 dysphagia occurred in 20% and 31% of patients with T1-T2 and T3-T4 tumours, respectively (p = 0.004). Corresponding values for N0-N1 and N2-N3 tumours were 20% and 39%, respectively (p = 0.002). The percentage of patients with Grade 3-7 dysphagia was 5%, 29%, 33% and 52% for oral cavity, laryngeal, oropharyngeal and hypopharyngeal tumours, respectively, (p = 0.002). Age and co-morbidity factors (e.g. diabetes, hypertension, coronary artery disease, peripheral vascular diseases and arthritis) did not appear to have an impact on swallowing in this limited retrospective study. Patients with locally advanced stages (T3-T4, N2-N3) are at risk of severe dysphagia. Patients with oral cavity tumours appear to be less at risk of dysphagia than those with tumours in different anatomic locations. The role of age and co-morbidity factors should be investigated in future prospective studies. PMID- 18508872 TI - Radiographical imaging of the normal anatomy and complications after gastric banding. AB - Laparoscopic adjustable gastric banding is a surgical procedure that is increasingly being performed for the treatment of morbid obesity. As with any intervention, gastric banding is not free from complications. Complications after gastric banding can be divided into early and late complications. Early complications include band malposition and perforation of the stomach. Late complications comprise pouch dilatation, intraluminal band penetration and oesophageal dilatation. Understanding the principles of the intervention is essential for both the interpretation of the resulting radiographical findings and the diagnosis of potential complications. We report on the normal anatomy and the most frequent complications seen after gastric banding. PMID- 18508873 TI - Whole-body PET/CT-mammography for staging breast cancer: initial results. AB - The purpose of this study was to evaluate the feasibility and utility of a dedicated positron emission tomography (PET)/CT protocol in breast cancer patients. 40 patients with suspected recurrent breast cancer underwent whole-body PET/CT in the supine position (SP) followed by PET/CT of the breasts and axillae in the prone position (PP) using a special positioning aid. PP and SP images were compared in terms of the tumour-to-thoracic-wall distance, tumour-to-skin distance and tumour volume, diameter, density, maximal standardized uptake value (SUV(max)) and localization. The size of axillary areas, the number of intra axillary lymph nodes, their transverse diameters, their SUV(max) and the number of distant metastases were compared between PP and SP images. Differences were tested for significance using the Student's t-test. All patients tolerated PP imaging well. Five locally recurrent breast cancers were detected, both in the SP and in the PP. Mean tumour-to-thoracic-wall distances (PP, 19 mm; SP, 8 mm; p = 0.003) and tumour-to-skin distances (PP, 10 mm; SP, 7 mm; p = 0.013) were significantly larger in the PP than in the SP. Potential thoracic wall or skin infiltration, as well as quadrant localization, were determined more easily in PP. The axillary area was wider in the PP when compared with SP (PP, 14.4 cm(2); SP, 10.6 cm(2); p<0.001). No other parameters were significantly different. In conclusion, a dedicated whole-body PET/CT examination, including PET/CT mammography, is feasible for clinical practice and may offer important information on the possible infiltration of a breast lesion into the adjacent thoracic wall and skin. Even though the axilla may be delineated more clearly in the PP, there seems to be no benefit with regard to N-staging. PMID- 18508874 TI - Cumulative patient effective dose in cardiology. AB - Medical radiation from X-rays and nuclear medicine is the largest non-natural (man-made) source of radiation exposure in Western countries. The aim of this study was to assess the individual cumulative effective dose in patients admitted to our cardiology ward. We collected a cumulative radiological history from a structured questionnaire and access to hospital records in 50 consecutive adult patients (36 males; age, 66.7+/-10.8 years) admitted to the Institute of Clinical Physiology in Pisa. The cumulative effective dose was assessed as an indicator of stochastic risk of cancer. We derived the effective dose for each individual examination from the Medical Imaging Guidelines of the European Commission (2001). On average, each patient underwent a median of 36 examinations (interquartile range, 23-46). The median cumulative effective dose was 60.6 mSv. Three types of procedures were responsible for approximately 86% of the total collective effective dose: (i) arteriography and interventional cardiology (12% of examinations, 48% of average dose per patient); (ii) nuclear medicine (5% of examinations, 21% of average dose per patient); and (iii) CT (4% of examinations, 17% of average dose per patient). The median estimated extra risk of cancer was approximately 1 in 200 exposed subjects. In conclusion, the average contemporary cardiological patient is exposed to a significant cumulative effective dose from diagnostic and therapeutic interventions. It is important to log cumulative dose for each patient at the time of each examination. Every effort should be made to justify the indications and to optimize the doses. PMID- 18508875 TI - Acute pulmonary coccidioidomycosis: CT findings from 15 patients. AB - The aim of this study was to describe the CT scan abnormalities in 15 patients with acute pulmonary coccidioidomycosis. Retrospective analysis of chest CT scans from 15 patients with acute pulmonary coccidioidomycosis was performed. The final diagnosis included the finding of Coccidioides immitis in mycology and/or histopathology, complemented by serology. Two radiologists evaluated the CT scans to study the type, size, profusion and localization of the findings. The final decisions were defined by consensus. CT scans showed multiple bilateral nodules in 13 patients and solitary nodules associated with consolidation in 2 cases. The nodules had ill-defined contours, ranging from 0.5 cm to 3.0 cm in diameter, which were predominant in the lower lobes in 11 cases. Cavitation of nodules was observed in 13 cases and coalescence in 7. Nodule-associated abnormalities were found in 13 cases, comprising interlobular septal thickening (n = 7) and consolidations (n = 6). Other abnormalities included lymph node enlargement (n = 6) and small pleural effusion (n = 2). In conclusion, the main CT finding in patients with acute coccidioidomycosis was that of multiple nodules (0.5-3.0 cm) at the lungs bases; a significant proportion of the remaining cases also showed other abnormalities. A diagnosis of coccidioidomycosis must be considered in patients with multiple lung nodules that are either in, or have recently been transported to, areas of endemic mycosis. PMID- 18508876 TI - Kidney-specific expression of human organic cation transporter 2 (OCT2/SLC22A2) is regulated by DNA methylation. AB - Human organic cation transporter 2 (OCT2/SLC22A2), which is specifically expressed in the kidney, plays critical roles in the renal secretion of cationic compounds. Tissue expression and membrane localization of OCT2 are closely related to the tissue distribution, pharmacological effects, and/or adverse effects of its substrate drugs. However, the molecular mechanisms underlying the kidney-specific expression of OCT2 have not been elucidated. In the present study, therefore, we examined the contribution of DNA methylation of the promoter region for the OCT2 gene to its tissue-specific expression using human tissue samples. In vivo methylation status of the proximal promoter region of OCT2 and that of OCT1, a liver-specific organic cation transporter, were investigated by bisulfite sequencing using human genomic DNA extracted from the kidney and liver. All CpG sites in the OCT2 proximal promoter were hypermethylated in the liver, while hypomethylated in the kidney. On the other hand, the promoter region of OCT1 was hypermethylated in both the kidney and liver. The level of methylation of the OCT2 promoter was especially low at the CpG site in the E-box, the binding site of the basal transcription factor upstream stimulating factor (USF) 1. In vitro methylation of the OCT2 proximal promoter dramatically reduced the transcriptional activity, and an electrophoretic mobility shift assay showed that methylation at the E-box inhibited the binding of USF1. These results indicate that kidney-specific expression of human OCT2 is regulated by methylation of the proximal promoter region, interfering with the transactivation by USF1. PMID- 18508878 TI - Enhanced bladder capacity and reduced prostaglandin E2-mediated bladder hyperactivity in EP3 receptor knockout mice. AB - Nonsteroidal anti-inflammatory cyclooxygenase inhibitors that function to reduce prostaglandin E2 (PGE2) production have been widely reported as effective agents in models of urinary bladder overactivity. We therefore investigated a potential role for the PGE2 receptor, EP3, in urinary bladder function by performing conscious, freely moving cystometry on EP3 receptor knockout (KO) mice. EP3 KO mice demonstrated an enhanced bladder capacity compared with wild-type (WT) mice ( approximately 185% of WT) under control conditions, based on larger voided and infused bladder volumes. Infusion of the EP3 receptor agonist GR63799X into the bladder of WT mice reduced the bladder capacity. This was ineffective in EP3 KO mice that demonstrated a time-dependent increase in bladder capacity with GR63799X, an effect similar to that observed with vehicle in both genotypes. In addition, infusion of PGE2 into WT mice induced bladder overactivity, an effect that was significantly blunted in the EP3 KO mice. The data reported here provide the first evidence supporting a functional role for EP3 receptors in normal urinary bladder function and implicate EP3 as a contributor to bladder overactivity during pathological conditions of enhanced PGE2 production, as reported previously in overactive bladder patients. PMID- 18508877 TI - Regulation of the epithelial sodium channel by membrane trafficking. AB - The epithelial Na(+) channel (ENaC) is a major regulator of salt and water reabsorption in a number of epithelial tissues. Abnormalities in ENaC function have been directly linked to several human disease states including Liddle's syndrome, psuedohypoaldosteronism, and cystic fibrosis and may be implicated in states as diverse as salt-sensitive hypertension, nephrosis, and pulmonary edema. ENaC activity in epithelial cells is highly regulated both by open probability and number of channels. Open probability is regulated by a number of factors, including proteolytic processing, while ENaC number is regulated by cellular trafficking. This review discusses current understanding of apical membrane delivery, cell surface stability, endocytosis, retrieval, and recycling of ENaC and the molecular partners that have so far been shown to participate in these processes. We review known sites and mechanisms of hormonal regulation of trafficking by aldosterone, vasopressin, and insulin. While many details of the regulation of ENaC trafficking remain to be elucidated, knowledge of these mechanisms may provide further insights into ENaC activity in normal and disease states. PMID- 18508879 TI - Ureter obstruction alters expression of renal acid-base transport proteins in rat kidney. AB - Urinary tract obstruction impairs renal function and is often associated with a urinary acidification defect caused by diminished net H+ secretion and/or HCO3- reabsorption. To identify the molecular mechanisms of these defects, protein expression of key acid-base transporters were examined along the renal nephron and collecting duct of kidneys from rats subjected to 24-h bilateral ureteral obstruction (BUO), 4 days after release of BUO (BUO-R), or BUO-R rats with experimentally induced metabolic acidosis (BUO-A). Semiquantitative immunoblotting revealed that BUO caused a significant reduction in the expression of the type 3 Na+/H+ exchanger (NHE3) in the cortex (21 +/- 4%), electrogenic Na+/HCO3- cotransporter (NBC1; 71 +/- 5%), type 1 bumetanide-sensitive Na+-K+-2Cl cotransporter (NKCC2; 3 +/- 1%), electroneutral Na+/HCO3- cotransporter (NBCn1; 46 +/- 7%), and anion exchanger (pendrin; 87 +/- 2%). The expression of H+-ATPase increased in the inner medullary collecting duct (152 +/- 13%). These changes were confirmed by immunocytochemistry. In BUO-R rats, there was a persistent downregulation of all the acid-base transporters including H+-ATPase. Two days of NH4Cl loading reduced plasma pH and HCO3- levels in BUO-A rats. The results demonstrate that the expression of multiple renal acid-base transporters are markedly altered in response to BUO, which may be responsible for development of metabolic acidosis and contribute to the urinary acidification defect after release of the obstruction. PMID- 18508880 TI - Covalent binding and tissue distribution/retention assessment of drugs associated with idiosyncratic drug toxicity. AB - Bioactivation of a drug to a reactive metabolite and its covalent binding to cellular macromolecules is believed to be involved in clinical adverse events, including idiosyncratic drug toxicities (IDTs). For the interpretation of the covalent binding data in terms of risk assessment, the in vitro and in vivo covalent binding data of a variety of drugs associated with IDTs or not were determined. Most of the "problematic" drugs, including "withdrawn" and "warning" drugs, exhibit higher human liver microsome (HLM) in vitro covalent binding yields than the "safe" drugs. Although some of the problematic drugs that are known to undergo bioactivation other than cytochrome P450-mediated oxidation exhibited only trace levels of HLM covalent binding like safe drugs, a rat in vivo covalent binding study could assess the bioactivation of such drugs. Furthermore, the tissue distribution/retention of the drugs was also examined by rat autoradiography (ARG). The residual radioactivity in the liver observed at 72 or 168 h postdose was found to be well correlated with the rat in vivo covalent binding to liver proteins; thus, the in vivo covalent binding yields of the drugs could be extrapolated from the retention profiles observed by means of ARG. Long term retention of radioactivity in the bone marrow was observed with some drugs associated with severe agranulocytosis, suggesting a spatial relationship between the toxicity profile and drug distribution/retention. Taken together, the covalent binding and tissue distribution/retention data of the various marketed drugs obtained in the present study should be quite informative for the interpretation of data in terms of risk assessment. PMID- 18508881 TI - Effect of interleukin-2 pretreatment on paclitaxel absorption and tissue disposition after oral and intravenous administration in mice. AB - The aim of the present study was to investigate the effects of recombinant interleukin (rIL)-2 treatment on paclitaxel (PLX) pharmacokinetics in the plasma and tissue of Lewis lung carcinoma-bearing mice (lung tissues and s.c. tumors). PLX pharmacokinetics studies were conducted after oral and i.v. administration of 15 and 4 mg/kg, respectively, either alone or after 3 days of rIL-2 pretreatment. The noncompartmental approach was used to determine the mean pharmacokinetic parameters using WinNonlin software (Pharsight, Mountain View, CA). The influence of rIL-2 pretreatment on physiological P-glycoprotein (P-gp) expression in lung and intestine was investigated by Western blot analysis. After oral administration of PLX, areas under the curve (AUC) in plasma, lung, and s.c. tumors were significantly higher (2.98, 2.66, and 3.41-fold, respectively) in the rIL-2 + PLX group as compared with the PLX group. However, no significant effect of rIL-2 pretreatment was observed in plasma or lung following i.v. administration of PLX. PLX AUC in s.c. tumors was significantly higher (1.37 fold) with rIL-2 pretreatment as compared with the PLX-alone group after i.v. injection. Pretreatment with rIL-2 appeared to have no effect on PLX plasma terminal half-life when PLX was administered orally or i.v. However, prolongation of PLX terminal half-life estimated from lung and s.c. tumors data had been observed. Increased PLX tissue absorption in the rIL-2-pretreated group may be explained by a decrease of P-gp expression in the intestines and lung or decreased functionality due to rIL-2. Oral administration allowed the targeted tissues a much higher PLX exposure as compared with i.v. administration. PMID- 18508882 TI - Dengue virus replicon expressing the nonstructural proteins suffices to enhance membrane expression of HLA class I and inhibit lysis by human NK cells. AB - Many viruses escape the cellular immune response by downregulating cell surface expression of major histocompatibility complex (MHC) class I molecules. However, infection of cells with flaviviruses can upregulate the expression of these molecules. In this study we analyzed the expression of MHC class I in K562 and THP-1 human cell lines that were stably transfected with self-replicating subgenomic dengue virus RNA (replicons) and express all the dengue virus nonstructural proteins together. We show that MHC class I expression is upregulated in the dengue virus replicon-expressing cells and that the binding of natural killer (NK) inhibitory receptors to these cells is augmented. This upregulation results in reduced susceptibility of the dengue virus replicon expressing cells to NK lysis, indicating a possible mechanism for evasion of the dengue virus from NK cell recognition. Visualizing MHC class I expression in replicon-containing K562 and THP-1 cells by confocal microscopy demonstrated aggregation of MHC class I molecules on the cell surface. Finally, replicon expressing K562 cells manifested increased TAP (transporter associated with antigen processing) and LMP (low-molecular-mass protein) gene transcription, while replicon-expressing THP-1 cells manifested increased NF-kappaB activity and MHC class I transcription. We suggest that expression of dengue virus nonstructural proteins is sufficient to induce MHC class I upregulation through both TAP-dependent and -independent mechanisms. Additionally, aggregation of MHC class I molecules on the cell membrane also contributes to significantly higher binding of low-affinity NK inhibitory receptors, resulting in lower sensitivity to lysis by NK cells. PMID- 18508883 TI - Dysregulation of TLR3 impairs the innate immune response to West Nile virus in the elderly. AB - West Nile virus (WNV), a mosquito-borne flavivirus, has recently emerged in North America, and the elderly are particularly susceptible to severe neurological disease and death from infection with this virus. We have investigated the innate immune response of primary human macrophages to WNV in vitro and have found significant differences between the responsiveness of macrophages derived from younger donors and that from older donors. Binding of the glycosylated WNV envelope protein to the C-type lectin dendritic cell-specific intercellular adhesion molecule 3 (ICAM3) grabbing nonintegrin (DC-SIGN) leads to a reduction in the expression of Toll-like receptor 3 (TLR3) in macrophages from young donors via the signal transducer and activator of transcription 1 (STAT1)-mediated pathway. This signaling is impaired in the elderly, and the elevated levels of TLR3 result in an elevation of cytokine levels. This alteration of the innate immune response with aging may contribute to the permeability of the blood-brain barrier and suggests a possible mechanism for the increased severity of WNV infection in older individuals. PMID- 18508884 TI - Human immunodeficiency virus type 1 infection increases the in vivo capacity of peripheral monocytes to cross the blood-brain barrier into the brain and the in vivo sensitivity of the blood-brain barrier to disruption by lipopolysaccharide. AB - Human immunodeficiency virus type 1 (HIV-1), introduced into the brain by HIV-1 infected monocytes which migrate across the blood-brain barrier (BBB), infects resident macrophages and microglia and initiates a process that causes HIV-1 associated neurocognitive disorders. The mechanism by which HIV-1 infection circumvents the BBB-restricted passage of systemic leukocytes into the brain and disrupts the integrity of the BBB is not known. Circulating lipopolysaccharide (LPS), which can compromise the integrity of the BBB, is significantly increased in HIV-1-infected individuals. We hypothesized that HIV-1 infection increases monocyte capacity to migrate across the BBB, which is further facilitated by a compromise of BBB integrity mediated by the increased systemic LPS levels present in HIV-1-infected individuals. To investigate this possibility, we examined the in vivo BBB migration of monocytes derived from our novel mouse model, JR CSF/EYFP mice, which are transgenic for both a long terminal repeat-regulated full-length infectious HIV-1 provirus and ROSA-26-regulated enhanced yellow fluorescent protein. We demonstrated that JR-CSF/EYFP mouse monocytes displayed an increased capacity to enter the brain by crossing either an intact BBB or a BBB whose integrity was partially compromised by systemic LPS. We also demonstrated that the JR-CSF mouse BBB was more susceptible to disruption by systemic LPS than the control wild-type mouse BBB. These results demonstrated that HIV-1 infection increased the ability of monocytes to enter the brain and increased the sensitivity of the BBB to disruption by systemic LPS, which is elevated in HIV-1-infected individuals. These mice represent a new in vivo system for studying the mechanism by which HIV-1-infected monocytes migrate into the brain. PMID- 18508885 TI - Different mechanisms of cell entry by human-pathogenic Old World and New World arenaviruses. AB - The Old World arenavirus Lassa virus (LASV) is the causative agent of severe viral hemorrhagic fever (VHF) in humans and is the most prevalent human pathogen among arenaviruses. The present study investigated the largely unknown mechanisms of cell entry of LASV, a process know to be mediated solely by the virus envelope glycoprotein (GP). To circumvent biosafety restrictions associated with the use of live LASV, we used reverse genetics to generate a recombinant variant of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) expressing the LASV GP (rLCMV-LASVGP). The rescued rLCMV-LASVGP grew to titers comparable to that of LCMV and showed the receptor binding characteristics of LASV. We used rLCMV-LASVGP to characterize the cellular mechanisms of LASV entry in the context of a productive arenavirus infection. The kinetics of pH-dependent membrane fusion of rLCMV-LASVGP resembled those of the human-pathogenic New World arenavirus Junin virus (JUNV) and other enveloped viruses that use clathrin mediated endocytosis for entry. However, rLCMV-LASVGP entered cells predominantly via a clathrin-, caveolin-, and dynamin-independent endocytotic pathway similar to the one recently described for LCMV. Productive infection of rLCMV-LASVGP was only mildly affected by a dominant negative mutant of Rab5 and was independent of Rab7, suggesting an unusual mechanism of delivery to endosomes. In addition, rLCMV-LASVGP infection was independent of actin but required intact microtubules. Our data indicate that LASV enters cells via a pathway distinct from the one used by human-pathogenic New World arenaviruses. PMID- 18508886 TI - Asparagine 631 variants of the chicken Mx protein do not inhibit influenza virus replication in primary chicken embryo fibroblasts or in vitro surrogate assays. AB - Whether chicken Mx inhibits influenza virus replication is an important question with regard to strategies aimed at enhancing influenza resistance in domestic flocks. The Asn631 polymorphism of the chicken Mx protein found in the Shamo (SHK) chicken line was previously reported to be crucial for the antiviral activity of this highly polymorphic chicken gene. Our aims were to determine whether cells from commercial chicken lines containing Asn631 alleles were resistant to influenza virus infection and to investigate the effects that other polymorphisms might have on Mx function. Unexpectedly, we found that the Asn631 genotype had no impact on multicycle replication of influenza virus (A/WSN/33 [H1N1]) in primary chicken embryo fibroblast lines. Furthermore, expression of the Shamo (SHK) chicken Mx protein in transfected 293T cells did not inhibit viral gene expression (A/PR/8/34 [H1N1], A/Duck/England/62 [H4N6], and A/Duck/Singapore/97 [H5N3]). Lastly, in minireplicon systems (A/PR/8/34 and A/Turkey/England/50-92/91 [H5N1]), which were highly sensitive to inhibition by the murine Mx1 and human MxA proteins, respectively, Shamo chicken Mx also proved ineffective in the context of avian as well as mammalian cell backgrounds. Our findings demonstrate that Asn631 chicken Mx alleles do not inhibit influenza virus replication of the five strains tested here and efforts to increase the frequency of Asn631 alleles in commercial chicken populations are not warranted. Nevertheless, chicken Mx variants with anti-influenza activity might still exist. The flow cytometry and minireplicon assays described herein could be used as efficient functional screens to identify such active chicken Mx alleles. PMID- 18508887 TI - Virological consequences of early events following cell-cell contact between human immunodeficiency virus type 1-infected and uninfected CD4+ cells. AB - Human immunodeficiency virus type 1 (HIV-1)-infected cells transmit viral products to uninfected CD4(+) cells very rapidly. However, the natures of the transmitted viral products and the mechanism of transmission, as well as the relative virological consequences, have not yet been fully clarified. We studied the virological events occurring a few hours after contact between HIV-1-infected and uninfected CD4(+) cells using a coculture cell system in which the virus expression in target cells could be monitored through the induction of a green fluorescent protein reporter gene driven by HIV-1 long terminal repeats. Within 16 h of coculture, we observed two phenomena not related to the cell-free virus infection, i.e., the formation of donor-target cell fusions and a fusion independent internalization of viral particles likely occurring at least in part through intercellular connections. Both events depended on the expression of Env and CD4 in donor and target cells, respectively, whereas the HIV-1 internalization required clathrin activity in target cells. Importantly, both phenomena were also observed in cocultures of primary CD4(+) lymphocytes, while primary macrophages supported only HIV-1 endocytosis. By investigating the virological consequences of these events, we noticed that while fused cells released infectious HIV-1 particles, albeit with reduced efficiency compared with donor cells, no virus expression was detectable upon HIV-1 endocytosis in target cells. In sum, the HIV-1 transmission following contact between an HIV-1-infected and an uninfected CD4(+) cell can occur through different mechanisms, leading to distinguishable virological outcomes. PMID- 18508888 TI - Reactive-site cleavage residues confer target specificity to baculovirus P49, a dimeric member of the P35 family of caspase inhibitors. AB - Baculovirus proteins P49 and P35 are potent suppressors of apoptosis in diverse organisms. Although related, P49 and P35 inhibit initiator and effector caspases, respectively, during infection of permissive insect cells. The molecular basis of this novel caspase specificity is unknown. To advance strategies for selective inhibition of the cell death caspases, we investigated biochemical differences between these baculovirus substrate inhibitors. We report here that P49 and P35 use similar mechanisms for stoichiometric inhibition that require caspase cleavage of their reactive site loops (RSL) and chemical contributions of a conserved N-terminal cysteine to stabilize the resulting inhibitory complex. Our data indicated that P49 functions as a homodimer that simultaneously binds two caspases. In contrast, P35 is a monomeric, monovalent inhibitor. P49 and P35 also differ in their RSL caspase recognition sequences. We tested the role of the P(4) P(1) recognition motif for caspase specificity by monitoring virus-induced proteolytic processing of Sf-caspase-1, the principal effector caspase of the host insect Spodoptera frugiperda. When P49's TVTD recognition motif was replaced with P35's DQMD motif, P49 was impaired for inhibition of the initiator caspase that cleaves and activates pro-Sf-caspase-1 and instead formed a stable inhibitory complex with active Sf-caspase-1. In contrast, the effector caspase specificity of P35 was unaltered when P35's DQMD motif was replaced with TVTD. We concluded that the TVTD recognition motif is required but not sufficient for initiator caspase inhibition by P49. Our findings demonstrate a critical role for the P(4)-P(1) recognition site in caspase specificity by P49 and P35 and indicate that additional determinants are involved in target selection. PMID- 18508889 TI - The first Chinese porcine sapovirus strain that contributed to an outbreak of gastroenteritis in piglets. PMID- 18508890 TI - Deoxynucleoside triphosphate incorporation mechanism of foamy virus (FV) reverse transcriptase: implications for cell tropism of FV. AB - Here, we investigated the pre-steady-state deoxynucleoside triphosphate (dNTP) incorporation kinetics of primate foamy virus (PFV) reverse transcriptase (RT) in comparison with those of HIV-1 and MuLV RTs. PFV RT displayed a drastic reduction in primer extension at low dNTP concentrations where HIV-1 RT remains highly active, indicating a low dNTP binding affinity in the case of PFV RT. Indeed, kinetic analysis showed that, as observed with MuLV RT, PFV RT exhibits approximately 10 to 80 times lower dNTP binding affinity than HIV-1 RT. These three RTs, however, show similar catalytic activities. In conclusion, PFV RT displays mechanistic distinctions in comparison to HIV-1 RT and shares close similarity to MuLV RT. PMID- 18508891 TI - Identification of residues outside of the receptor binding domain that influence the infectivity and tropism of porcine endogenous retrovirus. AB - Identification of determinants of human tropism of porcine endogenous retrovirus (PERV) is critical to understanding the risk of transmission of PERV to recipients of porcine xenotransplantation products. Previously, we showed that a chimeric envelope cDNA encoding the 360 N-terminal residues of the human-tropic PERV envelope class A (PERV-A) SU and the 130 C-terminal residues of the pig tropic PERV-C SU and all of TM (PERV-A/C) showed a 100-fold decrease in infectivity titer on human cells (M. Gemeniano, O. Mpanju, D. R. Salomon, M. V. Eiden, and C. A. Wilson, Virology 346:108-117, 2006). To identify residues important for human cell infection, we performed site-directed mutagenesis on each of the nine residues, singly or in combination, that distinguish the C terminal region of PERV-C from PERV-A. Of the nine amino acids, two single-amino acid substitutions, Q374R and I412V, restored the infectivity of human cells to the chimeric PERV-A/C to a titer equivalent to that of PERV-A. In contrast, PERV A/C mutant envelope Q439P resulted in undetectable infection of human cells and an approximately 1,000-fold decrease in control pig cells. Mutation of K441R rescued mutants that carried Q439P, suggesting an incompatibility between the proline residue at this position and the presence of KK in the proteolytic cleavage signal. We confirmed this incompatibility with vectors carrying PERV-A envelope mutant R462K that were also rendered noninfectious. Finally, tropism of vectors carrying PERV-C envelope mutants with only four amino acid changes in the C terminus of PERV-C envelope, NHRQ436YNRP plus K441R, was shifted to one similar to that of PERV-A. Our results show an important and previously unrecognized role for infectivity and tropism for residues at the C terminus of SU. PMID- 18508892 TI - Visualization of the externalized VP2 N termini of infectious human parvovirus B19. AB - The structures of infectious human parvovirus B19 and empty wild-type particles were determined by cryoelectron microscopy (cryoEM) to 7.5-A and 11.3-A resolution, respectively, assuming icosahedral symmetry. Both of these, DNA filled and empty, wild-type particles contain a few copies of the minor capsid protein VP1. Comparison of wild-type B19 with the crystal structure and cryoEM reconstruction of recombinant B19 particles consisting of only the major capsid protein VP2 showed structural differences in the vicinity of the icosahedral fivefold axes. Although the unique N-terminal region of VP1 could not be visualized in the icosahedrally averaged maps, the N terminus of VP2 was shown to be exposed on the viral surface adjacent to the fivefold beta-cylinder. The conserved glycine-rich region is positioned between two neighboring, fivefold symmetrically related VP subunits and not in the fivefold channel as observed for other parvoviruses. PMID- 18508893 TI - Cryoelectron microscopy map of Atadenovirus reveals cross-genus structural differences from human adenovirus. AB - A three-dimensional (3D) cryoelectron microscopy reconstruction of the prototype Atadenovirus (OAdV [an ovine adenovirus isolate]) showing information at a 10.6-A resolution (0.5 Fourier shell correlation) was derived by single-particle analysis. This is the first 3D structure solved for any adenovirus that is not a Mastadenovirus, allowing cross-genus comparisons between structures and the assignment of genus-specific capsid proteins. Viable OAdV mutants that lacked the genus-specific LH3 and p32k proteins in purified virions were also generated. Negatively stained 3D reconstructions of these mutants were used to identify the location of protein LH3 and infer that of p32k within the capsid. The key finding was that LH3 is a critical protein that holds the outer capsid of the virus together. In its absence, the outer viral capsid is unstable. LH3 is located in the same position among the hexon subunits as its protein IX equivalent from mastadenoviruses but sits on top of the hexon trimers, forming prominent "knobs" on the virion surface that visually distinguish OAdV from other known AdVs. Electron density was also assigned to hexon and penton subunits and to proteins IIIa and VIII. There was good correspondence between OAdV density and human AdV hexon structures, which also validated the significant differences that were observed between the penton base protein structures. PMID- 18508894 TI - NS3 helicase domains involved in infectious intracellular hepatitis C virus particle assembly. AB - A mutation within subdomain 1 of the hepatitis C virus (HCV) NS3 helicase (NS3 Q221L) (M. Yi, Y. Ma, J. Yates, and S. M. Lemon, J. Virol. 81:629-638, 2007) rescues a defect in production of infectious virus by an intergenotypic chimeric RNA (HJ3). Although NS3-Gln-221 is highly conserved across HCV genotypes, the Leu 221 substitution had no effect on RNA replication or NS3-associated enzymatic activities. However, while transfection of unmodified HJ3 RNA failed to produce either extracellular or intracellular infectious virus, transfection of HJ3 RNA containing the Q221L substitution (HJ3/QL) resulted in rapid accumulation of intracellular infectious particles with release into extracellular fluids. In the absence of the Q221L mutation, both NS5A and NS3 were recruited to core protein on the surface of lipid droplets, but there was no assembly of core into high density, rapidly sedimenting particles. Further analysis demonstrated that a Q221N mutation minimally rescued virus production and led to a second-site I399V mutation in subdomain 2 of the helicase. Similarly, I399V alone allowed only low level virus production and led to selection of an I286V mutation in subdomain 1 of the helicase which fully restored virus production, confirming the involvement of both major helicase subdomains in the assembly process. Thus, multiple mutations in the helicase rescue a defect in an early-intermediate step in virus assembly that follows the recruitment of NS5A to lipid droplets and precedes the formation of dense intracellular viral particles. These data reveal a previously unsuspected role for the NS3 helicase in early virion morphogenesis and provide a new perspective on HCV assembly. PMID- 18508895 TI - Interspecies transmission of simian foamy virus in a natural predator-prey system. AB - Simian foamy viruses (SFV) are ancient retroviruses of primates and have coevolved with their host species for as many as 30 million years. Although humans are not naturally infected with foamy virus, infection is occasionally acquired through interspecies transmission from nonhuman primates. We show that interspecies transmissions occur in a natural hunter-prey system, i.e., between wild chimpanzees and colobus monkeys, both of which harbor their own species specific strains of SFV. Chimpanzees infected with chimpanzee SFV strains were shown to be coinfected with SFV from colobus monkeys, indicating that apes are susceptible to SFV superinfection, including highly divergent strains from other primate species. PMID- 18508896 TI - Reduction in severity of a herpes simplex virus type 1 murine infection by treatment with a ribozyme targeting the UL20 gene RNA. AB - Hammerhead ribozymes were designed to target mRNA of several essential herpes simplex virus type 1 (HSV-1) genes. A ribozyme specific for the late gene U(L)20 was packaged in an adenovirus vector (Ad-U(L)20 Rz) and evaluated for its capacity to inhibit the viral replication of several HSV-1 strains, including that of the wild-type HSV-1 (17syn+ and KOS) and several acycloguanosine resistant strains (PAAr5, tkLTRZ1, and ACGr4) in tissue culture. The Ad-U(L)20 Rz was also tested for its ability to block an HSV-1 infection, using the mouse footpad model. Mouse footpads were treated with either the Ad-U(L)20 Rz or an adenoviral vector expressing green fluorescent protein (Ad-GFP) and then infected immediately thereafter with 10(4) PFU of HSV-1 strain 17syn+. Ad-U(L)20 ribozyme treatment consistently led to a 90% rate of protection for mice from lethal HSV-1 infection, while the survival rate in the control groups was less than 45%. Consistent with this protective effect, treatment with the Ad-U(L)20 Rz reduced the viral DNA load in the feet, the dorsal root ganglia, and the spinal cord relative to that of the Ad-GFP-treated animals. This study suggests that ribozymes targeting essential genes of the late kinetic class may represent a new therapeutic strategy for inhibiting HSV infection. PMID- 18508897 TI - Limited maintenance of vaccine-induced simian immunodeficiency virus-specific CD8 T-cell receptor clonotypes after virus challenge. AB - T-cell receptors (TCRs) govern the specificity, efficacy, and cross-reactivity of CD8 T cells. Here, we studied CD8 T-cell clonotypes from Mane-A*10(+) pigtail macaques responding to the simian immunodeficiency virus (SIV) Gag KP9 epitope in a setting of vaccination and subsequent viral challenge. We observed a diverse TCR repertoire after DNA, recombinant poxvirus, and live attenuated virus vaccination, with none of 59 vaccine-induced KP9-specific TCRs being identical between macaques. The KP9-specific TCR repertoires remained diverse after SIV or simian-human immunodeficiency virus challenge but, remarkably, exhibited substantially different clonotypic compositions compared to the corresponding populations prechallenge. Within serial samples from individual pigtail macaques, only a small subset (33.9%) of TCRs induced by vaccination were maintained or expanded after challenge. Most (66.1%) of the TCRs induced by vaccination were not detectable after challenge. Our results suggest that some CD8 T cells induced by vaccination are more efficient than others at responding to a viral challenge. These findings have implications for future AIDS virus vaccine studies, which should consider the "fitness" of vaccine-induced T cells in order to generate robust responses in the face of virus exposure. PMID- 18508898 TI - Atomic force microscopy investigation of vaccinia virus structure. AB - Vaccinia virus was treated in a controlled manner with various combinations of nonionic detergents, reducing agents, and proteolytic enzymes, and successive products of the reactions were visualized using atomic force microscopy (AFM). Following removal of the outer lipid/protein membrane, a layer 20 to 40 nm in thickness was encountered that was composed of fibrous elements which, under reducing conditions, rapidly decomposed into individual monomers on the substrate. Beneath this layer was the virus core and its prominent lateral bodies, which could be dissociated or degraded with proteases. The core, in addition to the lateral bodies, was composed of a thick, multilayered shell of proteins of diverse sizes and shapes. The shell, which was readily etched with proteases, was thoroughly permeated with pores, or channels. Prolonged exposure to proteases and reductants produced disgorgement of the viral DNA from the remainders of the cores and also left residual, flattened, protease-resistant sacs on the imaging substrate. The DNA was readily visualized by AFM, which revealed some regions to be "soldered" by proteins, others to be heavily complexed with protein, and yet other parts to apparently exist as bundled, naked DNA. Prolonged exposure to proteases deproteinized the DNA, leaving masses of extended, free DNA. Estimates of the interior core volume suggest moderate but not extreme compaction of the genome. PMID- 18508899 TI - Synthesis of genomic and subgenomic RNA in mosquito cells infected with two Sindbis virus nsP4 mutants: influence of intracellular nucleoside triphosphate concentrations. AB - Cells infected with Sindbis virus (SV) make two positive-strand RNAs, a genomic length RNA (G) RNA and a subgenomic (SG) RNA. In cells infected with SVstd, and in general in cells infected with wt alphaviruses, more SG RNA is made than G RNA. How the balance between synthesis of G RNA and SG RNA is regulated is not known. SVpzf and SVcpc are nsP4 mutants of SV which, in mosquito cells, make more G RNA than SG RNA. When low concentrations of pyrazofurin (inhibits the synthesis of UTP and CTP) were added to SVpzf-infected cells, the yield of virus was increased, and the ratio of SG/G RNA was changed from <1 to >1. These effects were reversed by uridine. In SVcpc-infected cells, but not in SVstd-infected cells, synthesis of viral RNA was inhibited by the addition of either uridine or cytidine, and viral yields were lowered. Our findings suggest that the activities of the viral RNA-synthesizing complexes in cells infected with SVpzf or SVcpc, in contrast to those in SVstd-infected cells, are sensitive to high concentrations of UTP or CTP. Using a cell-free system that synthesizes both SG and G RNA, we measured viral RNA synthesis as a function of the UTP/CTP concentrations. The results indicated that the presence of the SVpzf mutations in nsP4 and the SG promoter produced a pattern quite different from that seen with the SVstd nsP4 and SG promoter. As the UTP/CTP concentrations were increased, the SVpzf system, in contrast to the SVstd system, made more G RNA than SG RNA, reflecting the situation in cells infected with SVpzf. PMID- 18508900 TI - Induction of broad CD4+ and CD8+ T-cell responses and cross-neutralizing antibodies against hepatitis C virus by vaccination with Th1-adjuvanted polypeptides followed by defective alphaviral particles expressing envelope glycoproteins gpE1 and gpE2 and nonstructural proteins 3, 4, and 5. AB - Broad, multispecific CD4(+) and CD8(+) T-cell responses to the hepatitis C virus (HCV), as well as virus-cross-neutralizing antibodies, are associated with recovery from acute infection and may also be associated in chronic HCV patients with a favorable response to antiviral treatment. In order to recapitulate all of these responses in an ideal vaccine regimen, we have explored the use of recombinant HCV polypeptides combined with various Th1-type adjuvants and replication-defective alphaviral particles encoding HCV proteins in various prime/boost modalities in BALB/c mice. Defective chimeric alphaviral particles derived from the Sindbis and Venezuelan equine encephalitis viruses encoding either the HCV envelope glycoprotein gpE1/gpE2 heterodimer (E1E2) or nonstructural proteins 3, 4, and 5 (NS345) elicited strong CD8(+) T-cell responses but low CD4(+) T helper responses to these HCV gene products. In contrast, recombinant E1E2 glycoproteins adjuvanted with MF59 containing a CpG oligonucleotide elicited strong CD4(+) T helper responses but no CD8(+) T-cell responses. A recombinant NS345 polyprotein also stimulated strong CD4(+) T helper responses but no CD8(+) T-cell responses when adjuvanted with Iscomatrix containing CpG. Optimal elicitation of broad CD4(+) and CD8(+) T-cell responses to E1E2 and NS345 was obtained by first priming with Th1-adjuvanted proteins and then boosting with chimeric, defective alphaviruses expressing these HCV genes. In addition, this prime/boost regimen resulted in the induction of anti-E1E2 antibodies capable of cross-neutralizing heterologous HCV isolates in vitro. This vaccine formulation and regimen may therefore be optimal in humans for protection against this highly heterogeneous global pathogen. PMID- 18508901 TI - Murine gammaherpesvirus 68 open reading frame 75c tegument protein induces the degradation of PML and is essential for production of infectious virus. AB - Promyelocytic Leukemia nuclear body (PML NB) proteins mediate an intrinsic cellular host defense response against virus infections. Herpesviruses express proteins that modulate PML or PML-associated proteins by a variety of strategies, including degradation of PML or relocalization of PML NB proteins. The consequences of PML-herpesvirus interactions during infection in vivo have yet to be investigated in detail, largely because of the species-specific tropism of many human herpesviruses. Murine gammaherpesvirus 68 (gammaHV68) is emerging as a suitable model to study basic biological questions of virus-host interactions because it naturally infects mice. Therefore, we sought to determine whether gammaHV68 targets PML NBs as part of its natural life cycle. We found that gammaHV68 induces PML degradation through a proteasome-dependent mechanism and that loss of PML results in more robust virus replication in mouse fibroblasts. Surprisingly, gammaHV68-mediated PML degradation was mediated by the virion tegument protein ORF75c, which shares homology with the cellular formylglycinamide ribotide amidotransferase enzyme. In addition, we show that ORF75c is essential for production of infectious virus. ORF75 homologs are conserved in all rhadinoviruses but so far have no assigned functions. Our studies shed light on a potential role for this unusual protein in rhadinovirus biology and suggest that gammaHV68 will be a useful model for investigation of PML-herpesvirus interactions in vivo. PMID- 18508902 TI - Induction of plasma (TRAIL), TNFR-2, Fas ligand, and plasma microparticles after human immunodeficiency virus type 1 (HIV-1) transmission: implications for HIV-1 vaccine design. AB - The death of CD4(+) CCR5(+) T cells is a hallmark of human immunodeficiency virus (HIV) infection. We studied the plasma levels of cell death mediators and products--tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), Fas ligand, TNF receptor type 2 (TNFR-2), and plasma microparticles--during the earliest stages of infection following HIV type 1 (HIV-1) transmission in plasma samples from U.S. plasma donors. Significant plasma TRAIL level elevations occurred a mean of 7.2 days before the peak of plasma viral load (VL), while TNFR 2, Fas ligand, and microparticle level elevations occurred concurrently with maximum VL. Microparticles had been previously shown to mediate immunosuppressive effects on T cells and macrophages. We found that T-cell apoptotic microparticles also potently suppressed in vitro immunoglobulin G (IgG) and IgA antibody production by memory B cells. Thus, release of TRAIL during the onset of plasma viremia (i.e., the eclipse phase) in HIV-1 transmission may initiate or amplify early HIV-1-induced cell death. The window of opportunity for a HIV-1 vaccine is from the time of HIV-1 transmission until establishment of the latently infected CD4(+) T cells. Release of products of cell death and subsequent immunosuppression following HIV-1 transmission could potentially narrow the window of opportunity during which a vaccine is able to extinguish HIV-1 infection and could place severe constraints on the amount of time available for the immune system to respond to the transmitted virus. PMID- 18508903 TI - Suppression of astrovirus replication by an ERK1/2 inhibitor. AB - Human astroviruses are nonenveloped, positive-sense single-strand RNA viruses associated with self-limiting diarrhea. Although they are recognized as a leading cause of disease in young children, the cellular factors involved in astrovirus replication are not well defined. The extracellular signal-regulated kinase (ERK) pathway has been shown to regulate many viral infections, but its role during astrovirus infection is unknown. In this report, we show that astrovirus activates ERK1/2 early in infection independently of replication. Inhibition of ERK activation with U0126, a specific ERK inhibitor, significantly reduced viral production. Investigations into the mechanism of ERK1/2 regulation revealed that all steps of the viral life cycle, including early and late protein expression as well as subgenomic and genomic RNA transcription, were diminished during U0126 treatment of monolayers. These data support a role for ERK1/2 in a postattachment step, although the precise mechanism remains under investigation. PMID- 18508904 TI - Reovirus serotypes elicit distinctive patterns of recall immunity in humans. AB - Mammalian orthoreoviruses (reoviruses) are ubiquitous viral agents that infect cells in respiratory and enteric tracts. The frequency and nature of human cellular immunoregulatory responses against reovirus are unknown. Here we establish systems to detect and quantify reovirus-induced cytokine and chemokine recall responses using primary cultures of virus-infected peripheral blood mononuclear cells (PBMC) and two widely used reovirus serotypes, type 1 Lang (T1L) and type 3 Dearing (T3D) reexposure in vitro. In cultures from 44 healthy adults, reovirus induced exceptionally strong CD4 and CD8 T-cell-dependent gamma interferon (IFN-gamma) recall responses concomitant with intense interleukin 10 (IL-10) production. These responses were elicited independently of viral replication. Surprisingly, paired analyses of subject responses to these two common serotypes revealed that while both elicit intense Th1-dominated immunity, median T3D-driven responses were 2.2-fold weaker (P = 0.0004) than those elicited by T1L. Recall responses evoked by these viral serotypes differed markedly in their mechanism of regulation. T3D IL-10 and IFN-gamma responses were CD4 and CD8 dependent and blocked by interfering with CD86 costimulation but were CD80 independent. T1L responses were consistently CD28 and CD80/86 independent. Thus, despite extensive genetic and morphological similarities between reovirus serotypes, the nature and intensity of the human recall responses as well as the control mechanisms regulating them are clearly distinct. PMID- 18508905 TI - The Whole-genome sequencing of the obligate intracellular bacterium Orientia tsutsugamushi revealed massive gene amplification during reductive genome evolution. AB - Scrub typhus ('Tsutsugamushi' disease in Japanese) is a mite-borne infectious disease. The causative agent is Orientia tsutsugamushi, an obligate intracellular bacterium belonging to the family Rickettsiaceae of the subdivision alpha Proteobacteria. In this study, we determined the complete genome sequence of O. tsutsugamushi strain Ikeda, which comprises a single chromosome of 2 008 987 bp and contains 1967 protein coding sequences (CDSs). The chromosome is much larger than those of other members of Rickettsiaceae, and 46.7% of the sequence was occupied by repetitive sequences derived from an integrative and conjugative element, 10 types of transposable elements, and seven types of short repeats of unknown origins. The massive amplification and degradation of these elements have generated a huge number of repeated genes (1196 CDSs, categorized into 85 families), many of which are pseudogenes (766 CDSs), and also induced intensive genome shuffling. By comparing the gene content with those of other family members of Rickettsiacea, we identified the core gene set of the family Rickettsiaceae and found that, while much more extensive gene loss has taken place among the housekeeping genes of Orientia than those of Rickettsia, O. tsutsugamushi has acquired a large number of foreign genes. The O. tsutsugamushi genome sequence is thus a prominent example of the high plasticity of bacterial genomes, and provides the genetic basis for a better understanding of the biology of O. tsutsugamushi and the pathogenesis of 'Tsutsugamushi' disease. PMID- 18508906 TI - Force deficits and breakage rates after single lengthening contractions of single fast fibers from unconditioned and conditioned muscles of young and old rats. AB - The deficit in force generation is a measure of the magnitude of damage to sarcomeres caused by lengthening contractions of either single fibers or whole muscles. In addition, permeabilized single fibers may suffer breakages. Our goal was to understand the interaction between breakages and force deficits in "young" and "old" permeabilized single fibers from control muscles of young and old rats and "conditioned" fibers from muscles that completed a 6-wk program of in vivo lengthening contractions. Following single lengthening contractions of old control fibers compared with young-control fibers, the twofold greater force deficits at a 10% strain support the concept of an age-related increase in the susceptibility of fibers to mechanical damage. In addition, the much higher breakage rates for old fibers at all strains tested indicate an increase with aging in the number of fibers at risk of being severely injured during any given stretch. Following the 6-wk program of lengthening contractions, young conditioned fibers and old-conditioned fibers were not different with respect to force deficit or the frequency of breakages. A potential mechanism for the increased resistance to stretch-induced damage of old-conditioned fibers is that, through intracellular damage and subsequent degeneration and regeneration, weaker sarcomeres were replaced by stronger sarcomeres. These data indicate that, despite the association of high fiber breakage rates and large force deficits with aging, the detrimental characteristics of old fibers were improved by a conditioning program that altered both sarcomeric characteristics as well as the overall structural integrity of the fibers. PMID- 18508907 TI - Small mouse cholangiocytes proliferate in response to H1 histamine receptor stimulation by activation of the IP3/CaMK I/CREB pathway. AB - Cholangiopathies are characterized by the heterogeneous proliferation of different-sized cholangiocytes. Large cholangiocytes proliferate by a cAMP dependent mechanism. The function of small cholangiocytes may depend on the activation of inositol trisphosphate (IP(3))/Ca(2+)-dependent signaling pathways; however, data supporting this speculation are lacking. Four histamine receptors exist (HRH1, HRH2, HRH3, and HRH4). In several cells: 1) activation of HRH1 increases intracellular Ca(2+) concentration levels; and 2) increased [Ca(2+)](i) levels are coupled with calmodulin-dependent stimulation of calmodulin-dependent protein kinase (CaMK) and activation of cAMP-response element binding protein (CREB). HRH1 agonists modulate small cholangiocyte proliferation by activation of IP(3)/Ca(2+)-dependent CaMK/CREB. We evaluated HRH1 expression in cholangiocytes. Small and large cholangiocytes were stimulated with histamine trifluoromethyl toluidide (HTMT dimaleate; HRH1 agonist) for 24-48 h with/without terfenadine, BAPTA/AM, or W7 before measuring proliferation. Expression of CaMK I, II, and IV was evaluated in small and large cholangiocytes. We measured IP(3), Ca(2+) and cAMP levels, phosphorylation of CaMK I, and activation of CREB (in the absence/presence of W7) in small cholangiocytes treated with HTMT dimaleate. CaMK I knockdown was performed in small cholangiocytes stimulated with HTMT dimaleate before measurement of proliferation and CREB activity. Small and large cholangiocytes express HRH1, CaMK I, and CaMK II. Small (but not large) cholangiocytes proliferate in response to HTMT dimaleate and are blocked by terfenadine (HRH1 antagonist), BAPTA/AM, and W7. In small cholangiocytes, HTMT dimaleate increased IP(3)/Ca(2+) levels, CaMK I phosphorylation, and CREB activity. Gene knockdown of CaMK I ablated the effects of HTMT dimaleate on small cholangiocyte proliferation and CREB activation. The IP(3)/Ca(2+)/CaMK I/CREB pathway is important in the regulation of small cholangiocyte function. PMID- 18508908 TI - Abnormal sarcoplasmic reticulum Ca2+-sequestering properties in skeletal muscle in chronic obstructive pulmonary disease. AB - The objective of this study was to investigate the hypothesis that alterations in sarcoplasmic reticulum (SR) Ca(2+)-cycling properties would occur in skeletal muscle in patients with moderate to severe chronic obstructive pulmonary disease (COPD). To investigate this hypothesis, tissue samples were obtained from the vastus lateralis of 8 patients with COPD [age 65.6 +/- 3.2 yr; forced expiratory volume in 1 s (FEV(1))/forced vital capacity (FVC) = 44 +/- 2%; mean +/- SE] and 10 healthy age-matched controls (CON, age 67.5 +/- 2.5 yr; FEV(1)/FVC = 77 +/- 2%), and homogenates were analyzed for a wide range of SR properties. Compared with CON, COPD displayed (in mumol.g protein(-1).min(-1)) a 16% lower maximal Ca(2+)-ATPase activity [maximal velocity (V(max)), 158 +/- 10 vs. 133 +/- 7, P < 0.05] and a 17% lower Ca(2+) uptake (4.65 +/- 0.039 vs. 3.85 +/- 0.26, P < 0.05) that occurred in the absence of differences in Ca(2+) release. The lower V(max) in COPD was also accompanied by an 11% lower (P < 0.05) Ca(2+) sensitivity, as measured by the Hill coefficient (defined as the relationship between Ca(2+) ATPase activity and free cytosolic Ca(2+) concentration for 10-90% V(max)). For the sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) isoforms, SERCA1a was 16% higher (P < 0.05) and SERCA2a was 14% lower (P < 0.05) in COPD. It is concluded that moderate to severe COPD results in abnormalities in SR Ca(2+)-ATPase properties that cannot be explained by changes in the SERCA isoform phenotypes. The reduced catalytic properties of SERCA in COPD suggest a disturbance in Ca(2+) cycling, possibly resulting in impairment in Ca(2+)-mediated mechanical function and/or second messenger regulated processes. PMID- 18508909 TI - MURC, a muscle-restricted coiled-coil protein, is involved in the regulation of skeletal myogenesis. AB - Skeletal myogenesis is a multistep process by which multinucleated mature muscle fibers are formed from undifferentiated, mononucleated myoblasts. However, the molecular mechanisms of skeletal myogenesis have not been fully elucidated. Here, we identified muscle-restricted coiled-coil (MURC) protein as a positive regulator of myogenesis. In skeletal muscle, MURC was localized to the cytoplasm with accumulation in the Z-disc of the sarcomere. In C2C12 myoblasts, MURC expression occurred coincidentally with myogenin expression and preceded sarcomeric myosin expression during differentiation into myotubes. RNA interference (RNAi)-mediated knockdown of MURC impaired differentiation in C2C12 myoblasts, which was accompanied by impaired myogenin expression and ERK activation. Overexpression of MURC in C2C12 myoblasts resulted in the promotion of differentiation with enhanced myogenin expression and ERK activation during differentiation. During injury-induced muscle regeneration, MURC expression increased, and a higher abundance of MURC was observed in immature myofibers compared with mature myofibers. In addition, ERK was activated in regenerating tissue, and ERK activation was detected in MURC-expressing immature myofibers. These findings suggest that MURC is involved in the skeletal myogenesis that results from modulation of myogenin expression and ERK activation. MURC may play pivotal roles in the molecular mechanisms of skeletal myogenic differentiation. PMID- 18508910 TI - Role of caveolin-1 in the regulation of lipoprotein metabolism. AB - Lipoprotein metabolism plays an important role in the development of several human diseases, including coronary artery disease and the metabolic syndrome. A good comprehension of the factors that regulate the metabolism of the various lipoproteins is therefore key to better understanding the variables associated with the development of these diseases. Among the players identified are regulators such as caveolins and caveolae. Caveolae are small plasma membrane invaginations that are observed in terminally differentiated cells. Their most important protein marker, caveolin-1, has been shown to play a key role in the regulation of several cellular signaling pathways and in the regulation of plasma lipoprotein metabolism. In the present paper, we have examined the role of caveolin-1 in lipoprotein metabolism using caveolin-1-deficient (Cav-1(-/-)) mice. Our data show that, while Cav-1(-/-) mice show increased plasma triglyceride levels, they also display reduced hepatic very low-density lipoprotein (VLDL) secretion. Additionally, we also found that a caveolin-1 deficiency is associated with an increase in high-density lipoprotein (HDL), and these HDL particles are enriched in cholesteryl ester in Cav-1(-/-) mice when compared with HDL obtained from wild-type mice. Finally, our data suggest that a caveolin-1 deficiency prevents the transcytosis of LDL across endothelial cells, and therefore, that caveolin-1 may be implicated in the regulation of plasma LDL levels. Taken together, our studies suggest that caveolin-1 plays an important role in the regulation of lipoprotein metabolism by controlling their plasma levels as well as their lipid composition. Thus caveolin-1 may also play an important role in the development of atherosclerosis. PMID- 18508911 TI - Satellite cell proliferation is reduced in muscles of obese Zucker rats but restored with loading. AB - The obese Zucker rat (OZR) is a model of metabolic syndrome, which has lower skeletal muscle size than the lean Zucker rat (LZR). Because satellite cells are essential for postnatal muscle growth, this study was designed to determine whether reduced satellite cell proliferation contributes to reduced skeletal mass in OZR vs. LZR. Satellite cell proliferation was determined by a constant-release 5-bromo-2-deoxyuridine (BrdU) pellet that was placed subcutaneously in each animal. Satellite cell proliferation, as determined by BrdU incorporation, was significantly attenuated in control soleus and plantaris muscles of the OZR compared with that shown in the LZR. To determine whether this attenuation of satellite cell activity could be rescued in OZR muscles, soleus and gastrocnemius muscles were denervated, placing a compensatory load on the plantaris muscle. In the LZR and the OZR after 21 days of loading, increases of approximately 25% and approximately 30%, respectively, were shown in plantaris muscle wet weight compared with that shown in the contralateral control muscle. The number of BrdU positive nuclei increased similarly in loaded plantaris muscles from LZR and OZR. Myogenin, MyoD, and Akt protein expressions were lower in control muscles of OZR than in those of the LZR, but they were all elevated to similar levels in the loaded plantaris muscles of OZR and LZR. These data indicate that metabolic syndrome may reduce satellite cell proliferation, and this may be a factor that contributes to the reduced mass in control muscles of OZR; however, satellite cell proliferation can be restored with compensatory loading in OZR. PMID- 18508912 TI - Activation of hepatocytes by extracellular heat shock protein 72. AB - Heat shock protein (HSP) 72 is released by cells during stress and injury. HSP-72 also stimulates the release of cytokines in macrophages by binding to Toll-like receptors (TLR) 2 and 4. Circulating levels of HSP-72 increase during hepatic ischemia-reperfusion injury. The role of extracellular HSP-72 (eHSP-72) in the injury response to ischemia-reperfusion is unknown. Therefore, the objective of the present study was to determine whether eHSP-72 has any direct effects on hepatocytes. Primary mouse hepatocytes were treated with purified human recombinant HSP-72. Conditioned media were evaluated by ELISA for the cytokines, TNF-alpha, IL-6, and macrophage inflammatory protein 2 (MIP-2). Stimulation of hepatocytes with eHSP-72 did not induce production of TNFalpha or IL-6 but resulted in dose-dependent increases in MIP-2 production. To evaluate the pathway responsible for this response, expression of TLR2 and TLR4 was confirmed on hepatocytes by immunohistochemistry. Hepatocyte production of MIP-2 was significantly decreased in hepatocytes obtained from TLR2 or TLR4 knockout mice. MIP-2 production was found to be partially dependent on NF-kappaB because inhibition of NF-kappaB with Bay 11-7085 significantly decreased eHSP-72-induced MIP-2 production. Inhibitors of p38 mitogen-activated protein kinase or c-Jun NH(2)-terminal kinase had no effect on production of MIP-2 induced by eHSP-72. The data suggest that eHSP-72 binds to TLR2 and TLR4 on hepatocytes and signals through NF-kappaB to increase MIP-2 production. The fact that eHSP-72 did not increase TNF-alpha or IL-6 production may be indicative of a highly regulated signaling pathway downstream from TLR. PMID- 18508913 TI - Chromatin remodeling complexes interact dynamically with a glucocorticoid receptor-regulated promoter. AB - Brahma (BRM) and Brahma-related gene 1 (BRG1) are the ATP-dependent catalytic subunits of the SWI/SNF family of chromatin-remodeling complexes. These complexes are involved in essential processes such as cell cycle, growth, differentiation, and cancer. Using imaging approaches in a cell line that harbors tandem repeats of stably integrated copies of the steroid responsive MMTV-LTR (mouse mammary tumor virus-long terminal repeat), we show that BRG1 and BRM are recruited to the MMTV promoter in a hormone-dependent manner. The recruitment of BRG1 and BRM resulted in chromatin remodeling and decondensation of the MMTV repeat as demonstrated by an increase in the restriction enzyme accessibility and in the size of DNA fluorescence in situ hybridization (FISH) signals. This chromatin remodeling event was concomitant with an increased occupancy of RNA polymerase II and transcriptional activation at the MMTV promoter. The expression of ATPase deficient forms of BRG1 (BRG1-K-R) or BRM (BRM-K-R) inhibited the remodeling of local and higher order MMTV chromatin structure and resulted in the attenuation of transcription. In vivo photobleaching experiments provided direct evidence that BRG1, BRG1-K-R, and BRM chromatin-remodeling complexes have distinct kinetic properties on the MMTV array, and they dynamically associate with and dissociate from MMTV chromatin in a manner dependent on hormone and a functional ATPase domain. Our data provide a kinetic and mechanistic basis for the BRG1 and BRM chromatin-remodeling complexes in regulating gene expression at a steroid hormone inducible promoter. PMID- 18508914 TI - Retrotranslocation of prion proteins from the endoplasmic reticulum by preventing GPI signal transamidation. AB - Neurodegeneration in diseases caused by altered metabolism of mammalian prion protein (PrP) can be averted by reducing PrP expression. To identify novel pathways for PrP down-regulation, we analyzed cells that had adapted to the negative selection pressure of stable overexpression of a disease-causing PrP mutant. A mutant cell line was isolated that selectively and quantitatively routes wild-type and various mutant PrPs for ER retrotranslocation and proteasomal degradation. Biochemical analyses of the mutant cells revealed that a defect in glycosylphosphatidylinositol (GPI) anchor synthesis leads to an unprocessed GPI-anchoring signal sequence that directs both ER retention and efficient retrotranslocation of PrP. An unprocessed GPI signal was sufficient to impart ER retention, but not retrotranslocation, to a heterologous protein, revealing an unexpected role for the mature domain in the metabolism of misprocessed GPI-anchored proteins. Our results provide new insights into the quality control pathways for unprocessed GPI-anchored proteins and identify transamidation of the GPI signal sequence as a step in PrP biosynthesis that is absolutely required for its surface expression. As each GPI signal sequence is unique, these results also identify signal recognition by the GPI-transamidase as a potential step for selective small molecule perturbation of PrP expression. PMID- 18508915 TI - Differences in regulation of Drosophila and vertebrate integrin affinity by talin. AB - Integrin-mediated cell adhesion is essential for development of multicellular organisms. In worms, flies, and vertebrates, talin forms a physical link between integrin cytoplasmic domains and the actin cytoskeleton. Loss of either integrins or talin leads to similar phenotypes. In vertebrates, talin is also a key regulator of integrin affinity. We used a ligand-mimetic Fab fragment, TWOW-1, to assess talin's role in regulating Drosophila alphaPS2 betaPS affinity. Depletion of cellular metabolic energy reduced TWOW-1 binding, suggesting alphaPS2 betaPS affinity is an active process as it is for vertebrate integrins. In contrast to vertebrate integrins, neither talin knockdown by RNA interference nor talin head overexpression had a significant effect on TWOW-1 binding. Furthermore, replacement of the transmembrane or talin-binding cytoplasmic domains of alphaPS2 betaPS with those of human alphaIIb beta3 failed to enable talin regulation of TWOW-1 binding. However, substitution of the extracellular and transmembrane domains of alphaPS2 betaPS with those of alphaIIb beta3 resulted in a constitutively active integrin whose affinity was reduced by talin knockdown. Furthermore, wild-type alphaIIb beta3 was activated by overexpression of Drosophila talin head domain. Thus, despite evolutionary conservation of talin's integrin/cytoskeleton linkage function, talin is not sufficient to regulate Drosophila alphaPS2 betaPS affinity because of structural features inherent in the alphaPS2 betaPS extracellular and/or transmembrane domains. PMID- 18508916 TI - P4-ATPase requirement for AP-1/clathrin function in protein transport from the trans-Golgi network and early endosomes. AB - Drs2p is a resident type 4 P-type ATPase (P4-ATPase) and potential phospholipid translocase of the trans-Golgi network (TGN) where it has been implicated in clathrin function. However, precise protein transport pathways requiring Drs2p and how it contributes to clathrin-coated vesicle budding remain unclear. Here we show a functional codependence between Drs2p and the AP-1 clathrin adaptor in protein sorting at the TGN and early endosomes of Saccharomyces cerevisiae. Genetic criteria indicate that Drs2p and AP-1 operate in the same pathway and that AP-1 requires Drs2p for function. In addition, we show that loss of AP-1 markedly increases Drs2p trafficking to the plasma membrane, but does not perturb retrieval of Drs2p from the early endosome back to the TGN. Thus AP-1 is required at the TGN to sort Drs2p out of the exocytic pathway, presumably for delivery to the early endosome. Moreover, a conditional allele that inactivates Drs2p phospholipid translocase (flippase) activity disrupts its own transport in this AP-1 pathway. Drs2p physically interacts with AP-1; however, AP-1 and clathrin are both recruited normally to the TGN in drs2Delta cells. These results imply that Drs2p acts independently of coat recruitment to facilitate AP-1/clathrin coated vesicle budding from the TGN. PMID- 18508917 TI - A role for soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex dimerization during neurosecretion. AB - The interactions underlying the cooperativity of soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) complexes during neurotransmission are not known. Here, we provide a molecular characterization of a dimer formed between the cytoplasmic portions of neuronal SNARE complexes. Dimerization generates a two-winged structure in which the C termini of cytosolic SNARE complexes are in apposition, and it involves residues from the vesicle associated SNARE synaptobrevin 2 that lie close to the cytosol-membrane interface within the full-length protein. Mutation of these residues reduces stability of dimers formed between SNARE complexes, without affecting the stability of each individual SNARE complex. These mutations also cause a corresponding decrease in the ability of botulinum toxin-resistant synaptobrevin 2 to rescue regulated exocytosis in toxin-treated neuroendocrine cells. Moreover, such synaptobrevin 2 mutants give rise to a dominant-negative inhibition of exocytosis. These data are consistent with an important role for SNARE complex dimers in neurosecretion. PMID- 18508918 TI - Atg8 controls phagophore expansion during autophagosome formation. AB - Autophagy is a potent intracellular degradation process with pivotal roles in health and disease. Atg8, a lipid-conjugated ubiquitin-like protein, is required for the formation of autophagosomes, double-membrane vesicles responsible for the delivery of cytoplasmic material to lysosomes. How and when Atg8 functions in this process, however, is not clear. Here we show that Atg8 controls the expansion of the autophagosome precursor, the phagophore, and give the first real time, observation-based temporal dissection of the autophagosome formation process. We demonstrate that the amount of Atg8 determines the size of autophagosomes. During autophagosome biogenesis, Atg8 forms an expanding structure and later dissociates from the site of vesicle formation. On the basis of the dynamics of Atg8, we present a multistage model of autophagosome formation. This model provides a foundation for future analyses of the functions and dynamics of known autophagy-related proteins and for screening new genes. PMID- 18508919 TI - Saccharomyces cerevisiae Rot1 is an essential molecular chaperone in the endoplasmic reticulum. AB - Molecular chaperones prevent aggregation of denatured proteins in vitro and are thought to support folding of diverse proteins in vivo. Chaperones may have some selectivity for their substrate proteins, but knowledge of particular in vivo substrates is still poor. We here show that yeast Rot1, an essential, type-I ER membrane protein functions as a chaperone. Recombinant Rot1 exhibited antiaggregation activity in vitro, which was partly impaired by a temperature sensitive rot1-2 mutation. In vivo, the rot1-2 mutation caused accelerated degradation of five proteins in the secretory pathway via ER-associated degradation, resulting in a decrease in their cellular levels. Furthermore, we demonstrate a physical and probably transient interaction of Rot1 with four of these proteins. Collectively, these results indicate that Rot1 functions as a chaperone in vivo supporting the folding of those proteins. Their folding also requires BiP, and one of these proteins was simultaneously associated with both Rot1 and BiP, suggesting that they can cooperate to facilitate protein folding. The Rot1-dependent proteins include a soluble, type I and II, and polytopic membrane proteins, and they do not share structural similarities. In addition, their dependency on Rot1 appeared different. We therefore propose that Rot1 is a general chaperone with some substrate specificity. PMID- 18508920 TI - Xenopus TACC3/maskin is not required for microtubule stability but is required for anchoring microtubules at the centrosome. AB - Members of the transforming acidic coiled coil (TACC) protein family are emerging as important mitotic spindle assembly proteins in a variety of organisms. The molecular details of how TACC proteins function are unknown, but TACC proteins have been proposed to recruit microtubule-stabilizing proteins of the tumor overexpressed gene (TOG) family to the centrosome and to facilitate their loading onto newly emerging microtubules. Using Xenopus egg extracts and in vitro assays, we show that the Xenopus TACC protein maskin is required for centrosome function beyond recruiting the Xenopus TOG protein XMAP215. The conserved C-terminal TACC domain of maskin is both necessary and sufficient to restore centrosome function in maskin-depleted extracts, and we provide evidence that the N terminus of maskin inhibits the function of the TACC domain. Time-lapse video microscopy reveals that microtubule dynamics in Xenopus egg extracts are unaffected by maskin depletion. Our results provide direct experimental evidence of a role for maskin in centrosome function and suggest that maskin is required for microtubule anchoring at the centrosome. PMID- 18508921 TI - DHHC2 affects palmitoylation, stability, and functions of tetraspanins CD9 and CD151. AB - Although palmitoylation markedly affects tetraspanin protein biochemistry and functions, relevant palmitoylating enzymes were not known. There are 23 mammalian "DHHC" (Asp-His-His-Cys) proteins, which presumably palmitoylate different sets of protein substrates. Among DHHC proteins tested, DHHC2 best stimulated palmitoylation of tetraspanins CD9 and CD151, whereas inactive DHHC2 (containing DH-->AA or C-->S mutations within the DHHC motif) failed to promote palmitoylation. Furthermore, DHHC2 associated with CD9 and CD151, but not other cell surface proteins, and DHHC2 knockdown diminished CD9 and CD151 palmitoylation. Knockdown of six other Golgi-resident DHHC proteins (DHHC3, -4, 8, -17, -18, and -21) had no effect on CD9 or CD151. DHHC2 selectively affected tetraspanin palmitoylation, but not the palmitoylations of integrin beta4 subunit and bulk proteins visible in [(3)H]palmitate-labeled whole cell lysates. DHHC2 dependent palmitoylation also had multiple functional effects. First, it promoted physical associations between CD9 and CD151, and between alpha3 integrin and other proteins. Second, it protected CD151 and CD9 from lysosomal degradation. Third, the presence of DHHC2, but not other DHHC proteins, shifted cells away from a dispersed state and toward increased cell-cell contacts. PMID- 18508922 TI - Fas splicing regulation during early apoptosis is linked to caspase-mediated cleavage of U2AF65. AB - U2 small nuclear ribonucleoprotein (snRNP) auxiliary factor 65 kDa (U2AF65) is an essential splicing factor in the recognition of the pre-mRNA 3' splice sites during the assembly of the splicing commitment complex. We report here that U2AF65 is proteolyzed during apoptosis. This cleavage is group I or III caspase dependent in a noncanonical single site localized around the aspartic acid(128) residue and leads to the separation of the N- and C-terminal parts of U2AF65. The U2AF65 N-terminal fragment mainly accumulates in the nucleus within nuclear bodies (nucleoli-like pattern) and to a much lesser extent in the cytoplasm, whereas the C-terminal fragment is found in the cytoplasm, even in localization studies on apoptosis induction. From a functional viewpoint, the N-terminal fragment promotes Fas exon 6 skipping from a reporter minigene, by acting as a dominant-negative version of U2AF65, whereas the C-terminal fragment has no significant effect. The dominant-negative behavior of the U2AF65 N-terminal fragment can be reverted by U2AF35 overexpression. Interestingly, U2AF65 proteolysis in Jurkat cells on induction of early apoptosis correlates with the down-regulation of endogenous Fas exon 6 inclusion. Thus, these results support a functional link among apoptosis induction, U2AF65 cleavage, and the regulation of Fas alternative splicing. PMID- 18508923 TI - The PHD domain of Np95 (mUHRF1) is involved in large-scale reorganization of pericentromeric heterochromatin. AB - Heterochromatic chromosomal regions undergo large-scale reorganization and progressively aggregate, forming chromocenters. These are dynamic structures that rapidly adapt to various stimuli that influence gene expression patterns, cell cycle progression, and differentiation. Np95-ICBP90 (m- and h-UHRF1) is a histone binding protein expressed only in proliferating cells. During pericentromeric heterochromatin (PH) replication, Np95 specifically relocalizes to chromocenters where it highly concentrates in the replication factories that correspond to less compacted DNA. Np95 recruits HDAC and DNMT1 to PH and depletion of Np95 impairs PH replication. Here we show that Np95 causes large-scale modifications of chromocenters independently from the H3:K9 and H4:K20 trimethylation pathways, from the expression levels of HP1, from DNA methylation and from the cell cycle. The PHD domain is essential to induce this effect. The PHD domain is also required in vitro to increase access of a restriction enzyme to DNA packaged into nucleosomal arrays. We propose that the PHD domain of Np95-ICBP90 contributes to the opening and/or stabilization of dense chromocenter structures to support the recruitment of modifying enzymes, like HDAC and DNMT1, required for the replication and formation of PH. PMID- 18508924 TI - Ubc4/5 and c-Cbl continue to ubiquitinate EGF receptor after internalization to facilitate polyubiquitination and degradation. AB - c-Cbl is the E3 ubiquitin ligase that ubiquitinates the epidermal growth factor (EGF) receptor (EGFR). On the basis of localization, knockdown, and in vitro activity analyses, we have identified the E2 ubiquitin-conjugating enzyme that cooperates with c-Cbl as Ubc4/5. Upon EGF stimulation, both Ubc4/5 and c-Cbl were relocated to the plasma membrane and then to Hrs-positive endosomes, strongly suggesting that EGFR continues to be ubiquitinated after internalization. Our time-course experiment showed that EGFR undergoes polyubiquitination, which seemed to be facilitated during the transport to Hrs-positive endosomes. Use of a conjugation-defective ubiquitin mutant suggested that receptor polyubiquitination is required for efficient interaction with Hrs and subsequent sorting to lysosomes. Abrupt inhibition of the EGFR kinase activity resulted in dissociation of c-Cbl from EGFR. Concomitantly, EGFR was rapidly deubiquitinated and its degradation was delayed. We propose that sustained tyrosine phosphorylation of EGFR facilitates its polyubiquitination in endosomes and counteracts rapid deubiquitination, thereby ensuring Hrs-dependent lysosomal sorting. PMID- 18508925 TI - The yeast GID complex, a novel ubiquitin ligase (E3) involved in the regulation of carbohydrate metabolism. AB - Glucose-dependent regulation of carbon metabolism is a subject of intensive studies. We have previously shown that the switch from gluconeogenesis to glycolysis is associated with ubiquitin-proteasome linked elimination of the key enzyme fructose-1,6-bisphosphatase. Seven glucose induced degradation deficient (Gid)-proteins found previously in a genomic screen were shown to form a complex that binds FBPase. One of the subunits, Gid2/Rmd5, contains a degenerated RING finger domain. In an in vitro assay, heterologous expression of GST-Gid2 leads to polyubiquitination of proteins. In addition, we show that a mutation in the degenerated RING domain of Gid2/Rmd5 abolishes fructose-1,6-bisphosphatase polyubiquitination and elimination in vivo. Six Gid proteins are present in gluconeogenic cells. A seventh protein, Gid4/Vid24, occurs upon glucose addition to gluconeogenic cells and is afterwards eliminated. Forcing abnormal expression of Gid4/Vid24 in gluconeogenic cells leads to fructose-1,6-bisphosphatase degradation. This suggests that Gid4/Vid24 initiates fructose-1,6-bisphosphatase polyubiquitination by the Gid complex and its subsequent elimination by the proteasome. We also show that an additional gluconeogenic enzyme, phosphoenolpyruvate carboxykinase, is subject to Gid complex-dependent degradation. Our study uncovers a new type of ubiquitin ligase complex composed of novel subunits involved in carbohydrate metabolism and identifies Gid4/Vid24 as a major regulator of this E3. PMID- 18508926 TI - DNA-ligase IV and DNA-protein kinase play a critical role in deficient caspases activation in apoptosis-resistant cancer cells by using doxorubicin. AB - Resistance toward cytotoxic drugs is one of the primary causes for therapeutic failure in cancer therapy. DNA repair mechanisms as well as deficient caspases activation play a critical role in apoptosis resistance of tumor cells toward anticancer drug treatment. Here, we discovered that deficient caspases activation in apoptosis-resistant cancer cells depends on DNA-ligase IV and DNA-protein kinase (DNA-PK), playing crucial roles in the nonhomologous end joining (NHEJ) pathway, which is the predominant pathway for DNA double-strand break repair (DNA DSB-repair) in mammalian cells. DNA-PK(+/+) as well as DNA-ligase IV (+/+) cancer cells were apoptosis resistant and deficient in activation of caspase-3, caspase 9, and caspase-8 and in cleavage of poly(ADP-ribose) polymerase after doxorubicin treatment. Inhibition of NHEJ by knocking out DNA-PK or DNA-ligase IV restored caspases activation and apoptosis sensitivity after doxorubicin treatment. In addition, inhibition of caspases activation prevented doxorubicin-induced apoptosis but could not prevent doxorubicin-induced DNA damage, indicating that induction of DNA damage is independent of caspases activation. However, caspases activation depends on induction of DNA damage left unrepaired by NHEJ-DNA-DSB repair. We conclude that DNA damage left unrepaired by DNA-ligase IV or DNA-PK might be the initiator for caspases activation by doxorubicin in cancer cells. Failure in caspases activation using doxorubicin depends on loss of DNA damage and is due to higher rates of NHEJ-DNA-DBS-repair. PMID- 18508927 TI - A novel juxtamembrane domain in tumor necrosis factor receptor superfamily molecules activates Rac1 and controls neurite growth. AB - Members of the tumor necrosis factor receptor (TNFR) superfamily control cell fate determination, including cell death and differentiation. Fas (CD95) is the prototypical "death receptor" of the TNFR superfamily and signals apoptosis through well established pathways. In the adult nervous system, Fas induces apoptosis in the context of neuropathology such as stroke or amyotrophic lateral sclerosis. However, during nervous system development, Fas promotes neurite growth and branching. The molecular mechanisms underlying Fas-induced process formation and branching have remained unknown to date. Here, we define the molecular pathway linking Fas to process growth and branching in cell lines and in developing neurons. We describe a new cytoplasmic membrane proximal domain (MPD) that is essential for Fas-induced process growth and that is conserved in members of the TNFR superfamily. We show that the Fas MPD recruits ezrin, a molecule that links transmembrane proteins to the cytoskeleton, and activates the small GTPase Rac1. Deletion of the MPD, but not the death domain, abolished Rac1 activation and process growth. Furthermore, an ezrin-derived inhibitory peptide prevented Fas-induced neurite growth in primary neurons. Our results define a new domain, topologically and functionally distinct from the death domain, which regulates neuritogenesis via recruitment of ezrin and activation of Rac1. PMID- 18508928 TI - MicroRNA-21 targets Sprouty2 and promotes cellular outgrowths. AB - The posttranscriptional regulator, microRNA-21 (miR-21), is up-regulated in many forms of cancer, as well as during cardiac hypertrophic growth. To understand its role, we overexpressed it in cardiocytes where it revealed a unique type of cell to-cell "linker" in the form of long slender outgrowths and branches. We subsequently confirmed that miR-21 directly targets and down-regulates the expression of Sprouty2 (SPRY2), an inhibitor of branching morphogenesis and neurite outgrowths. We found that beta-adrenergic receptor (betaAR) stimulation induces up-regulation of miR-21 and down-regulation of SPRY2 and is, likewise, associated with connecting cell branches. Knockdown of SPRY2 reproduced the branching morphology in cardiocytes, and vice versa, knockdown of miR-21 using a specific 'miRNA eraser' or overexpression of SPRY2 inhibited betaAR-induced cellular outgrowths. These structures enclose sarcomeres and connect adjacent cardiocytes through functional gap junctions. To determine how this aspect of miR 21 function translates in cancer cells, we knocked it down in colon cancer SW480 cells. This resulted in disappearance of their microvillus-like protrusions accompanied by SPRY2-dependent inhibition of cell migration. Thus, we propose that an increase in miR-21 enhances the formation of various types of cellular protrusions through directly targeting and down-regulating SPRY2. PMID- 18508930 TI - Influence of Mycobacterium bovis BCG vaccination on cellular immune response of guinea pigs challenged with Mycobacterium tuberculosis. AB - Mycobacterium bovis bacillus Calmette-Guerin (BCG) currently remains the only licensed vaccine for the prevention of tuberculosis. In this study, we used a newly described flow cytometric technique to monitor changes in cell populations accumulating in the lungs and lymph nodes of naive and vaccinated guinea pigs challenged by low-dose aerosol infection with virulent Mycobacterium tuberculosis. As anticipated, vaccinated guinea pigs controlled the growth of the challenge infection more efficiently than controls did. This early phase of bacterial control in immune animals was associated with increased accumulation of CD4 and CD8 T cells, including cells expressing the activation marker CD45, as well as macrophages expressing class II major histocompatibility complex molecules. As the infection continued, the numbers of T cells in the lungs of vaccinated animals waned, whereas the numbers of these cells expressing CD45 increased. Whereas BCG vaccination reduced the influx of heterophils (neutrophils) into the lungs, an early B-cell influx was observed in these vaccinated animals. Overall, vaccine protection was associated with reduced pathology and lung damage in the vaccinated animals. These data provide the first direct evidence that BCG vaccination accelerates the influx of protective T-cell and macrophage populations into the infected lungs, diminishes the accumulation of nonprotective cell populations, and reduces the severity of lung pathology. PMID- 18508931 TI - Development of an immunofluorescence assay using recombinant proteins expressed in insect cells to screen and confirm presence of human herpesvirus 8-specific antibodies. AB - Human herpesvirus 8 (HHV-8), or Kaposi's sarcoma (KS)-associated herpesvirus, has been linked to all forms of KS. The results of most current serological assays for the detection of HHV-8-specific antibodies have low levels of concordance among themselves. To establish a sensitive and specific testing strategy that can be used to screen for HHV-8-specific antibodies, three HHV-8 proteins, ORF65, ORF73, and K8.1A, were expressed by using baculoviral vectors in insect cells and incorporated into a monoclonal antibody-enhanced immunofluorescence assay (mIFA) termed the Sf9 three-antigen mIFA. The results obtained by this mIFA were compared to those obtained by a standard mIFA with an HHV-8-infected B-cell line (BC3 mIFA). Test sera were obtained from patients diagnosed with KS, human immunodeficiency virus type 1-infected patients at high risk for HHV-8 infection, and healthy controls from a local blood bank. The combined use of both assays had a sensitivity of 94% and a specificity of 96%. The performance of these two assays when they were used together indicates that they may be useful for the reliable detection of HHV-8-specific immunoglobulin G antibodies in a population. PMID- 18508932 TI - Comparison of three commercial immunoassays for detection of herpes simplex virus type 2 antibodies in commercial sex workers in Yunnan Province, China. AB - Five hundred commercial sex workers in China were tested for herpes simplex virus type 2 by three immunoassays and Western blotting. Sensitivities for the Focus, Kalon, and Biokit assays were 86.7%, 82.3%, and 34.9%, respectively, and specificities were 91.8%, 94.2%, and 60.1%, respectively. The Focus assay performed optimally at an index of 1.5 (95.2% sensitivity and 93.4% specificity), and the Kalon assay performed optimally at an index of 1.2 (93.3% sensitivity and 95.2% specificity). PMID- 18508933 TI - Molecular characterization of porcine picobirnaviruses and development of a specific reverse transcription-PCR assay. AB - The molecular characterization of partial- length genomic segment 2 of porcine picobirnavirus (PBV) strains and the development of a specific reverse transcription-PCR (RT-PCR) assay for detection of virus in feces are reported. Phylogenetic analysis indicated that the studied porcine isolates were more closely related (>85% identity) to human PBV belonging to genogroup I than to the other porcine PBV described so far. Analysis by RT-PCR and polyacrylamide gel electrophoresis of fecal samples collected in Venezuela and Argentina showed that PBV circulate at high frequencies in piglets. PMID- 18508934 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream isolates in urban Detroit. AB - To gain a better understanding of epidemiology of resistance in Staphylococcus aureus, we describe the molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream isolates in urban Detroit. Bloodstream isolates from July 2005 to February 2007 were characterized. Two hundred ten bloodstream isolates from 201 patients were evaluated. Patient characteristics were as follows: median age, 54 years; 56% male; and 71% African-American. Seventy-six percent of infections were health care associated, with 55% being community-onset infections and 21% hospital acquired, and 24% were community associated. The most common sources were skin/wound (25%), central venous catheters (24%), unknown source (20%), and endocarditis (9%). Ninety percent and 5% of isolates had a MIC of vancomycin of or=1.5 mg/liter. Results of pulsed-field gel electrophoresis showed 17 strain types. The predominant strains were USA100 (104 isolates) and USA300 (74 isolates). Forty nine percent of the isolates had staphylococcal cassette chromosome mec II, and 56% had agr II. All USA300 isolates were positive for the Panton-Valentine leukocidin toxin genes and agr I. Forty-seven percent of USA300 bloodstream infections were health care associated (35% community onset and 12% hospital onset). USA300 strains were more common in injection drug users with skin/wound as the predominant source of infection. Thirty percent of the USA100 strains were closely related to vancomycin-resistant Staphylococcus aureus isolates. The results of this study show that vancomycin MICs using automated dilution testing with Vitek-2 and E-test were highly discordant. Most methicillin-resistant S. aureus strains causing bacteremia are health care associated, commonly have MICs of vancomycin that are high within the susceptible range are not detected by routine automated dilution testing, and have significant diversity of molecular characteristics. USA100 strains that are closely related to vancomycin-resistant S. aureus (VRSA) isolates and USA300 strains are common as causes of both hospital and community-onset infection. Infection control measures should focus not only on prevention of the spread of community strains in the hospital but also prevention of the spread of hospital strains associated with VRSA into the community. PMID- 18508935 TI - Capsular polysaccharide synthesis regions in Klebsiella pneumoniae serotype K57 and a new capsular serotype. AB - Community-acquired pyogenic liver abscess caused by Klebsiella pneumoniae is an emerging infectious disease. We explored the capsular polysaccharide synthesis (cps) regions of three non-K1, non-K2 K. pneumoniae strains, A1142, A7754, and A1517, from Taiwanese patients experiencing pyogenic liver abscess. Two of the strains, A1142 and A7754, belonged to capsular serotype K57, while the third belonged to a new capsular serotype, different from the previously reported 77 serotypes. Deletion and complementation experiments suggested that a unique K57 gene, a homologue of wzy, was essential for K57 capsular synthesis and confirmed that this gene cluster was a genetic coding region for K57. Compared to K1 and K2 strains, the three strains were all serum sensitive, suggesting that host factors might also be involved in the three patients. PCR using primers from specific genes for K57 was more sensitive and specific than traditional serotyping. The remaining strain, A1517, did not react to the antisera from any of the 77 serotypes, and none of the 77 reference strains reacted to the serum against this strain. Moreover, PCR analyses using various primer pairs from the serotype specific open reading frames did not reveal cross-reactivity to any of the 77 reference strains, suggesting that this strain likely represents a new capsular type. We conclude that sequences from these two cps regions are very useful in detecting K57 and the new cps genotype. PMID- 18508937 TI - Multiplex real-time PCR assay for rapid identification of Mycobacterium tuberculosis complex members to the species level. AB - The species identification of members of the Mycobacterium tuberculosis complex is critical to the timely initiation of both appropriate antibiotic therapy and proper public health control measures. However, the current commercially available molecular assays identify mycobacteria only to the complex level and are unable to differentiate M. tuberculosis from the closely related M. bovis and M. bovis BCG. We describe here a rapid and robust two-step, multiplex, real-time PCR assay based on genomic deletions to definitively identify M. tuberculosis, M. bovis, M. bovis BCG, and other members of the complex. When tested against a panel of well-characterized mycobacterial reference strains, the assay was both sensitive and specific, correctly identifying all strains. We applied this assay to 60 clinical isolates previously identified as M. tuberculosis complex and found 57 M. tuberculosis isolates and 3 M. bovis BCG isolates from patients who had received intravesical BCG. Furthermore, analysis of 15 clinical specimens previously identified as M. bovis by spoligotyping revealed an isolate of M. tuberculosis that had been misidentified. We propose that this assay will allow the routine identification of M. tuberculosis complex members in the clinical laboratory. PMID- 18508936 TI - Opa protein repertoires of disease-causing and carried meningococci. AB - The meningococcal Opa proteins play an important role in pathogenesis by mediating invasion of human cells. The aim of this investigation was to determine whether carried and disease-associated meningococci possess different Opa repertoires and whether the diversity of these proteins is associated with clinical severity of disease. Opa repertoires in 227 disease-associated meningococci, isolated in the United Kingdom over a period of 6 years, were compared to the repertoires in 190 asymptomatically carried meningococci isolated in the United Kingdom from a contemporary, nonepidemic period. Multidimensional scaling (MDS) was employed to investigate the association between Opa repertoires and multilocus sequence typing (MLST) genotypes. Associations with clinical severity were also analyzed statistically. High levels of diversity were observed in opa alleles, variable regions, and repertoires, and MDS revealed that MLST genotypes were strongly associated with particular Opa repertoires. Individual Opa proteins or repertoires were not associated with clinical severity, though there was a trend toward an association with the opaD locus. Meningococcal Opa repertoire is strongly linked to MLST genotype irrespective of epidemiological sampling and therefore correlates with invasiveness. It is not, however, strongly associated with severity of meningococcal disease. PMID- 18508938 TI - Mycobacterium haemophilum epididymal abscess in a renal transplant patient. AB - Mycobacterium haemophilum is an established cause of cutaneous lesions in immunocompromised hosts. We report the first known case of epididymal abscess, which highlights the need to work up all specimens that are acid-fast bacillus positive for M. haemophilum from immunocompromised hosts, regardless of body site. PMID- 18508939 TI - Biological and molecular characteristics of Mycobacterium tuberculosis clinical isolates with low-level resistance to isoniazid in Japan. AB - We reevaluated the BACTEC MGIT 960 antimicrobial susceptibility testing system (MGIT 960 AST) by using 1,112 isolates of Mycobacterium tuberculosis. When the results of MGIT 960 AST were compared with that of the proportion method using Ogawa medium (Ogawa PM), discrepant results were obtained for 30 strains with isoniazid, all resistant by MGIT 960 AST but susceptible by Ogawa PM. For 93% of the strains that produced discrepant results, the MIC was 0.4 or 0.8 microg/ml, showing resistance by the proportion method using Middlebrook agar plates. Furthermore, it was also established by analyses of the katG and inhA genes that strains resistant only by MGIT 960 AST have a low level of isoniazid (INH) resistance, indicating that MGIT 960 AST is a reliable method. Ninety-six strains were resistant to 0.1 microg/ml INH by MGIT 960 AST. When they were divided into three groups, Low-S (susceptible at 0.2 microg/ml), Low-R (resistant at 0.2 microg/ml), and High-R (resistant at 1.0 microg/ml), by Ogawa PM, 43.3% of the Low-S strains had mutations in the promoter region of inhA and no mutations were detected in katG codon 315, while 61.7% of the High-R strains had katG codon 315 mutations or a gross deletion of katG. These results suggest that mutations in inhA are associated with low-level resistance to INH and katG codon 315 mutations are associated with high-level resistance to INH. In addition, the analyses demonstrated some relationship of mutations in the inhA gene with ethionamide resistance for the Low-S strains, but not for the High-R strains. PMID- 18508940 TI - Distribution, organization, and ecology of bacteria in chronic wounds. AB - Between 1 and 2% of the population in the developed world experiences a nonhealing or chronic wound characterized by an apparent arrest in a stage dominated by inflammatory processes. Lately, research groups have proposed that bacteria might be involved in and contribute to the lack of healing of these wounds. To investigate this, we collected and examined samples from chronic wounds obtained from 22 different patients, all selected because of suspicion of Pseudomonas aeruginosa colonization. These wound samples were investigated by standard culturing methods and peptide nucleic acid-based fluorescence in situ hybridization (PNA FISH) for direct identification of bacteria. By means of the culturing methods, Staphylococcus aureus was detected in the majority of the wounds, whereas P. aeruginosa was observed less frequently. In contrast, using PNA FISH, we found that a large fraction of the wounds contained P. aeruginosa. Furthermore, PNA FISH revealed the structural organization of bacteria in the samples. It appeared that P. aeruginosa aggregated as microcolonies imbedded in the matrix component alginate, which is a characteristic hallmark of the biofilm mode of growth. The present investigation suggests that bacteria present within these wounds tend to be aggregated in microcolonies imbedded in a self-produced matrix, characteristic of the biofilm mode of growth. Additionally, we must conclude that there exists no good correlation between bacteria detected by standard culturing methods and those detected by direct detection methods such as PNA FISH. This strongly supports the development of new diagnostic and treatment strategies for chronic wounds. PMID- 18508941 TI - Application of a novel, rapid, and sensitive oligonucleotide ligation assay for detection of cancer-predicting mutations in the precore and basal core promoter of hepatitis B virus. AB - Hepatocellular carcinoma (HCC) and cirrhosis are important causes of mortality worldwide. Persistent hepatitis B virus (HBV) infection is a major cause of these diseases. Double mutations in the basal core promoter (BCP) (A1762T and G1764A) and precore (pre-C) (G1896A) regions of the virus are associated with progression to HCC. The current study is aimed at developing a simple method for screening and detecting BCP and pre-C mutations in HBV carriers. We have developed and validated an oligonucleotide ligation assay (OLA) to detect point mutations in the HBV core gene. We have applied OLA methods to samples from HBV-infected carriers recruited from the Gambia Liver Cancer Study (GLCS) comprising asymptomatic HBsAg carriers, patients with cirrhosis, and patients with HCC. We observed an 89.3% and 95.8% concordance between the OLA and DNA sequencing for BCP and pre-C mutations, respectively. OLA detected the mutations in single strain infections and in infections with mixtures of wild-type and mutant viruses under conditions where sequencing detected only the single dominant strains. BCP mutations were detected in 75.7% of patients with advanced liver disease (cirrhosis/HCC) compared to 47.6% of asymptomatic carriers, while pre-C mutations were detected in 34.5% of advanced liver disease patients and in 47.6% of asymptomatic HBsAg carriers. There was a significant association between the presence of BCP mutations and advanced liver disease. In conclusion, OLA is a simple, economical, and reliable assay for detection of pre-C and BCP mutations. Its application can lead to improvement in diagnosis and clinical care in regions where HBV is endemic. PMID- 18508942 TI - Rapid, novel, specific, high-throughput assay for diagnosis of Loa loa infection. AB - The ability to diagnose Loa loa infection readily and accurately remains a demanding task. Among the available diagnostic methods, many are impractical for point-of-care field testing. To investigate whether luciferase immunoprecipitation systems (LIPS) can be used for rapid and specific diagnosis of L. loa infection, a LIPS assay was developed based on immunoglobulin G (IgG) and IgG4 subclass antibodies to a recombinant L. loa SXP-1 (designated LlSXP-1) antigen and tested with sera from healthy controls or patients with proven infection with L. loa, Mansonella perstans, Onchocerca volvulus, Strongyloides stercoralis, or Wuchereria bancrofti. A LIPS test measuring IgG antibody against LlSXP-1 readily differentiated L. loa-infected from uninfected patients and demonstrated markedly improved sensitivity and specificity compared with an LlSXP 1 IgG4-based enzyme-linked immunosorbent assay (67% sensitivity and 99% specificity). No significant immunoreactivity was observed with S. stercoralis infected sera, but a small number of patients infected with O. volvulus, M. perstans, or W. bancrofti showed positive immunoreactivity. Measuring anti-IgG4 specific antibodies to LlSXP-1 showed a significant correlation (r approximately 0.85; P < 0.00001) with the anti-IgG results but showed no advantage over measuring the total IgG response alone. In contrast, a rapid LIPS format (called QLIPS) in which the tests are performed in less than 15 minutes under nonequilibrium conditions significantly improved the specificity for cross reactive O. volvulus patient sera (100% sensitivity and 100% specificity). These results suggest that LIPS (and the even more rapid test QLIPS) represents a major advance in the ability to diagnose L. loa infection and may have future applications for point-of-care diagnostics. PMID- 18508943 TI - Identification of a novel Babesia sp. from a sable antelope (Hippotragus niger Harris, 1838). AB - Babesiosis in a sable antelope (Hippotragus niger Harris, 1838) was first reported in 1930; the parasite was named Babesia irvinesmithi. Recently, specimens from an adult sable that presented with a sudden onset of disease and that subsequently died during immobilization were submitted for molecular characterization. Microscopic examination of thin blood smears revealed the presence of small piroplasms. DNA was extracted from blood samples; the V4 variable region of the 18S rRNA gene was amplified and analyzed using the reverse line blot (RLB) assay. Amplicons did not hybridize with any of the Babesia or Theileria species-specific probes present on the blot and hybridized only with a Babesia or Theileria genus-specific probe, suggesting the presence of a novel species. The full-length 18S rRNA gene sequence was obtained and aligned with published sequences of related genera, and phylogenetic trees were constructed. Sequence similarity analyses indicated that a Babesia species, designated Babesia sp. (sable), was present. The sequence showed its highest similarity to B. orientalis and to an unnamed Babesia species previously detected in bovine samples. The latter was later established to be Babesia occultans. A Babesia sp. (sable)-specific RLB oligonucleotide probe was designed and used to screen 200 South African sable samples, but so far, no other sample has been found to be positive for the presence of Babesia sp. (sable) DNA. In summary, we identified a novel piroplasm parasite from a sable antelope that died from an unknown illness. While the parasite was observed in blood smears, there is no direct evidence that it was the cause of death. PMID- 18508944 TI - Genetic diversity and zoonotic potential of Cryptosporidium parvum causing foal diarrhea. AB - Cryptosporidium isolates from diarrheic foals in New Zealand (n = 9) were identified as C. parvum, subtyped at two polymorphic loci, and compared with human (n = 45) and bovine (n = 8) isolates. Foal C. parvum isolates were genetically diverse, markedly similar to human and bovine isolates, and carried GP60 IIaA18G3R1 alleles, indicating a zoonotic potential. PMID- 18508945 TI - Pauci-immune crescentic glomerulonephritis superimposed on diabetic glomerulosclerosis. AB - BACKGROUND AND OBJECTIVES: Pauci-immune necrotizing and crescentic glomerulonephritis (PNCGN) superimposed on diabetic glomerulosclerosis (DGS) is a rare occurrence. Only limited data on this dual glomerulopathy are available. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Twenty-three cases of PNCGN superimposed on DGS were identified from the archives of the Renal Pathology Laboratory of Columbia University. The clinical features, pathologic findings, and outcomes are described. RESULTS: The majority of patients were white, elderly, and had longstanding diabetes. Patients presented with acute renal failure and an active urine sediment. Antinuclear cytoplasmic autoantibody (ANCA) testing was positive by indirect immunofluorescence in 18 of 22 patients. Sixteen patients had a P-ANCA pattern, 9 of whom underwent further testing and were found to be MPO-ANCA positive by enzyme-linked immunosorbent assay. Among the two patients with C-ANCA by indirect immunofluorescence, enzyme-linked immunosorbent assay was performed in one and revealed PR3-ANCA. Eight patients had extrarenal manifestations of vasculitis, including 6 with pulmonary hemorrhage. At the time of presentation and renal biopsy, 11 patients required hemodialysis. The mean percentages of glomeruli with cellular crescents, fibrous crescents, and necrosis were 24.9, 8.4, and 12.9, respectively. Most patients were treated with cyclophosphamide and prednisone. At a mean follow-up of 14.6 mo (available in 21 patients), 8 patients had died and 8 of the remaining 13 patients had reached end stage renal disease. Correlates of end-stage renal disease were hemodialysis at presentation and the degree of DGS. CONCLUSIONS: PNCGN may occur superimposed on DGS. The prognosis for this dual glomerulopathy is dismal despite aggressive therapy. PMID- 18508946 TI - Effect of tonsillectomy plus steroid pulse therapy on clinical remission of IgA nephropathy: a controlled study. AB - BACKGROUND AND OBJECTIVES: Few well-designed investigations have examined how tonsillectomy plus steroid pulse therapy affects IgA nephropathy. A prospective, controlled study therefore was performed to compare the effects of combined therapy with those of steroid pulse alone in patients with IgA nephropathy. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Fifty-five patients were followed up for 54.0 +/- 21.2 mo. Thirty-five of them underwent tonsillectomy and steroid pulse therapy (group C), and 20 received steroid pulse monotherapy (group M). Both groups received methylprednisolone intravenously, followed by oral prednisolone (initial dosage 0.5 mg/kg per d) for 12 to 18 mo. Primary evaluation items were a 100% increase in serum creatinine from baseline levels or the disappearance of urinary protein (UP) and/or occult blood (UOB) indicating clinical remission. RESULTS: At 24 mo after the initial treatment, the ratios of the UP and UOB disappearance were higher in group C than in group M, and the therapeutic effect persisted until the final observation. None of group C achieved a 100% increase in serum creatinine from the baseline level, whereas one patient in group M developed ESRD during the observation period. The histologic findings of repeated biopsy specimens from 18 patients revealed that mesangial proliferation and IgA deposition were significantly more reduced in group C than in group M. The Cox regression model showed that the combined therapy was approximately six-fold more effective in causing the disappearance of UP than steroid pulse monotherapy. CONCLUSION: Tonsillectomy combined with steroid pulse treatment can induce clinical remission in patients with IgA nephropathy. PMID- 18508947 TI - Single-dose pharmacokinetics of roflumilast in children and adolescents. AB - Roflumilast is an orally administered phosphodiesterase 4 inhibitor that has potential for use in pediatric patients with asthma. The pharmacokinetics of roflumilast and roflumilast N-oxide were examined in adolescents and children with stable mild to moderate asthma in an open-label crossover study with age stratification and 2 treatment periods (100-microg dose in period 1, 250-microg dose in period 2) separated by a washout period. Plasma concentrations were measured by high-performance liquid chromatography tandem mass spectrometry. Pharmacokinetic parameters were determined using standard noncompartmental methods and compared between study groups and within the entire cohort. Roflumilast was well tolerated. Linear relationships were evident for dose and area under the plasma drug concentration-time curve extrapolated to infinity for both roflumilast (r(2) = 0.36, P < .01) and roflumilast N-oxide (r(2) = 0.39, P < .01). With the exception of dose-normalized maximum plasma concentration (mean 1.1 and 0.8 microg/L per 1 microg/kg dose for adolescents and children, respectively), pharmacokinetic parameters for roflumilast and roflumilast N-oxide were not different between age groups and were similar to adults. PMID- 18508948 TI - Absolute oral bioavailability and disposition of deferasirox in healthy human subjects. AB - Deferasirox is a novel iron chelator formulated as tablets for dispersion (suspension) for once-a-day oral administration. The current study evaluated the absolute bioavailability of a single 375-mg oral dose of deferasirox administered in the form of tablets compared with a 130-mg intravenous infusion of deferasirox. Since this was a first-in-man study using the deferasirox intravenous (IV) formulation, the safety and tolerability of the IV formulation was evaluated in a pilot phase with a lower dose (65 mg) in 3 subjects prior to the main phase. The main study phase consisted of 17 healthy male volunteers. Plasma concentrations of deferasirox were measured following each treatment, and pharmacokinetic parameters including absolute oral bioavailability were determined. Absolute oral bioavailability of the deferasirox tablets was 70% (90% confidence interval, 62%-80%). Deferasirox was characterized as having a low plasma clearance of 3.53 (+/- 0.87) L/h. A small volume of distribution of deferasirox at steady state (V(ss)) of 14.37 (+/-2.69 L) was determined, indicating a low tissue distribution. PMID- 18508949 TI - Pharmacokinetics, efficacy, and tolerability of eslicarbazepine acetate in children and adolescents with epilepsy. AB - This study investigates the pharmacokinetics of eslicarbazepine acetate (ESL), a new voltage-gated sodium channel blocker, in epileptic children aged 2 to 7 years (n = 11) and 7 to 11 years (n = 8) and adolescents aged 12 to 17 years (n = 10). The study explores ESL efficacy and tolerability. Patients were treated with ESL once-daily doses of 5 mg/kg/day on weeks 1 to 4, 15 mg/kg/day on weeks 5 to 8, and 30 mg/kg/day (or 1800 mg/day, whichever was less) on weeks 9 to 12. At the end of each 4-week period, a 24-hour pharmacokinetic profiling was performed. Similar to adults, ESL was rapidly metabolized to eslicarbazepine. In all age groups, eslicarbazepine peak concentrations were reached 0.5 hour to 3 hours after ESL dosing, and C(max) and AUC(0-24) were dose proportional. Eslicarbazepine C(max) was similar between age groups following administration of identical ESL dose/kg, but AUC(0-24) depended on age due to a faster plasma clearance of eslicarbazepine in younger children compared with adolescents. R licarbazepine and oxcarbazepine were minor metabolites. A dose-dependent decrease in seizure frequency was observed in children aged 2 to 7 years and adolescents aged 12 to 17 years but not in children aged 7 to 11 years. One patient in each group became seizure free. ESL was generally well tolerated. PMID- 18508950 TI - Multiple-dose pharmacokinetics, pharmacodynamics, and safety of taranabant, a novel selective cannabinoid-1 receptor inverse agonist, in healthy male volunteers. AB - Taranabant is a cannabinoid-1 receptor inverse agonist for the treatment of obesity. This study evaluated the safety, pharmacokinetics, and pharmacodynamics of taranabant (5, 7.5, 10, or 25 mg once daily for 14 days) in 60 healthy male subjects. Taranabant was rapidly absorbed, with a median t(max) of 1.0 to 2.0 hours and a t(1/2) of approximately 74 to 104 hours. Moderate accumulation was observed in C(max) (1.18- to 1.40-fold) and AUC(0-24 h) (1.5- to 1.8-fold) over 14 days for the 5-, 7.5-, and 10-mg doses, with an accumulation half-life ranging from 15 to 21 hours. Steady state was reached after 13 days. After multiple-dose administration, plasma AUC(0-24 h) and C(max) of taranabant increased dose proportionally (5-10 mg) and increased somewhat less than dose proportionally for 25 mg. Taranabant was generally well tolerated up to doses of 10 mg and exhibited multiple-dose pharmacokinetics consistent with once-daily dosing. PMID- 18508951 TI - Dose- and time-dependent pharmacokinetics of midostaurin in patients with diabetes mellitus. AB - Midostaurin is a novel potent inhibitor of both protein kinase C and the major receptor for vascular endothelial growth factor involved in angiogenesis, presenting a rationale for its use in diabetic retinopathy. This study evaluated the safety and pharmacokinetics of midostaurin following multiple oral doses of midostaurin for 28 days at 4 dose levels (25 mg bid, 50 mg bid, 75 mg bid, 75 mg tid), as well as a single oral 100-mg dose in patients with diabetes mellitus (n = 9-13 per dose cohort). Pharmacokinetic parameters were determined on days 1 and 28 based on the plasma concentrations of midostaurin and its metabolites, CGP62221 and CGP52421. The plasma exposures (C(max) and AUC(0-tau)) of midostaurin and metabolites increased less than proportionally over the dose range of 25 to 100 mg, showing a 2.2-fold increase after the first dose. Midostaurin concentrations increased during the first 3 to 6 days of dosing, then declined with time (by 30%-50%) until a steady state was achieved, representing an average accumulation factor (R) of 1.7. CGP62221 showed a similar concentration-time pattern as midostaurin (R = 2.5), but CGP52421 accumulated significantly (R = 18.8). A high-fat meal was found to significantly increase the C(max) and AUC(0-12 h) of midostaurin by 1.5-fold (P = .04) and 1.8-fold (P = .01), respectively, compared with taking the drug after an overnight fast. Midostaurin administered at 50 to 225 mg/day appeared to be generally safe in this group of patients. The most common treatment-related adverse events (eg, loose stools, nausea, vomiting, and headache) were found to be dose related, and the frequency increased markedly above the 150-mg/day dose level. PMID- 18508952 TI - Potential involvement of iron in the pathogenesis of peritoneal endometriosis. AB - The aim of this study is to review the current literature associating endometriosis with iron and to discuss the potential causes and consequences of iron overload in the pelvic cavity. Indeed, iron is essential for all living organisms. However, excess iron can result in toxicity and is associated with pathological disorders. In endometriosis patients, iron overload has been demonstrated in the different components of the peritoneal cavity (peritoneal fluid, endometriotic lesions, peritoneum and macrophages). Animal models allow us to gather essential information on the origin, metabolism and effect of iron overload in endometriosis, which may originate from erythrocytes carried into the pelvic cavity mainly by retrograde menstruation. Peritoneal macrophages play an important role in the degradation of these erythrocytes and in subsequent peritoneal iron metabolism. Iron overload could affect a wide range of mechanisms involved in endometriosis development, such as oxidative stress or lesion proliferation. In conclusion, excess iron accumulation can result in toxicity and may be one of the factors contributing to the development of endometriosis. Treatment with an iron chelator could thus be beneficial in endometriosis patients to prevent iron overload in the pelvic cavity, thereby diminishing its deleterious effect. PMID- 18508953 TI - The level of free intracellular zinc mediates programmed cell death/cell survival decisions in plant embryos. AB - Zinc is a potent regulator of programmed cell death (PCD) in animals. While certain, cell-type-specific concentrations of intracellular free zinc are required to protect cells from death, zinc depletion commits cells to death in diverse systems. As in animals, PCD has a fundamental role in plant biology, but its molecular regulation is poorly understood. In particular, the involvement of zinc in the control of plant PCD remains unknown. Here, we used somatic embryos of Norway spruce (Picea abies) to investigate the role of zinc in developmental PCD, which is crucial for correct embryonic patterning. Staining of the early embryos with zinc-specific molecular probes (Zinquin-ethyl-ester and Dansylaminoethyl-cyclen) has revealed high accumulation of zinc in the proliferating cells of the embryonal masses and abrupt decrease of zinc content in the dying terminally differentiated suspensor cells. Exposure of early embryos to a membrane-permeable zinc chelator N,N,N',N'-tetrakis(2 pyridylmethyl)ethylenediamine led to embryonic lethality, as it induced ectopic cell death affecting embryonal masses. This cell death involved the loss of plasma membrane integrity, metacaspase-like proteolytic activity, and nuclear DNA fragmentation. To verify the anti-cell death effect of zinc, we incubated early embryos with increased concentrations of zinc sulfate. Zinc supplementation inhibited developmental PCD and led to suppression of terminal differentiation and elimination of the embryo suspensors, causing inhibition of embryo maturation. Our data demonstrate that perturbation of zinc homeostasis disrupts the balance between cell proliferation and PCD required for plant embryogenesis. This establishes zinc as an important cue governing cell fate decisions in plants. PMID- 18508954 TI - The maize phytoene synthase gene family: overlapping roles for carotenogenesis in endosperm, photomorphogenesis, and thermal stress tolerance. AB - Carotenoids are essential for photosynthesis and photoprotection; they also serve as precursors to signaling molecules that influence plant development and biotic/abiotic stress responses. With potential to improve plant yield and nutritional quality, carotenoids are targets for metabolic breeding/engineering, particularly in the Poaceae (grass family), which includes the major food crops. Depending on genetic background, maize (Zea mays) endosperm carotenoid content varies, and therefore breeding-enhanced carotenoid levels have been of ongoing interest. The first committed step in the plastid-localized biosynthetic pathway is mediated by the nuclear-encoded phytoene synthase (PSY). The gene family in maize and other grasses contains three paralogs with specialized roles that are not well understood. Maize endosperm carotenoid accumulation requires PSY1 expression. A maize antibody was used to localize PSY1 to amyloplast envelope membranes and to determine PSY1 accumulation in relation to carotenoid accumulation in developing endosperm. To test when and if PSY transcript levels correlated with carotenoid content, advantage was taken of a maize germplasm diversity collection that exhibits genetic and chemical diversity. Total carotenoid content showed statistically significant correlation with endosperm transcript levels at 20 d after pollination for PSY1 but not PSY2 or PSY3. Timing of PSY1 transcript abundance, previously unknown, provides critical information for choosing breeding alleles or properly controlling introduced transgenes. PSY1 was unexpectedly found to have an additional role in photosynthetic tissue, where it was required for carotenogenesis in the dark and for heat stress tolerance. Leaf carotenogenesis was shown to require phytochrome-dependent and phytochrome independent photoregulation of PSY2 plus nonphotoregulated PSY1 expression. PMID- 18508955 TI - The Arabidopsis BRAHMA chromatin-remodeling ATPase is involved in repression of seed maturation genes in leaves. AB - Synthesis and accumulation of seed storage proteins (SSPs) is an important aspect of the seed maturation program. Genes encoding SSPs are specifically and highly expressed in the seed during maturation. However, the mechanisms that repress the expression of these genes in leaf tissue are not well understood. To gain insight into the repression mechanisms, we performed a genetic screen for mutants that express SSPs in leaves. Here, we show that mutations affecting BRAHMA (BRM), a SNF2 chromatin-remodeling ATPase, cause ectopic expression of a subset of SSPs and other embryogenesis-related genes in leaf tissue. Consistent with the notion that such SNF2-like ATPases form protein complexes in vivo, we observed similar phenotypes for mutations of AtSWI3C, a BRM-interacting partner, and BSH, a SNF5 homolog and essential SWI/SNF subunit. Chromatin immunoprecipitation experiments show that BRM is recruited to the promoters of a number of embryogenesis genes in wild-type leaves, including the 2S genes, expressed in brm leaves. Consistent with its role in nucleosome remodeling, BRM appears to affect the chromatin structure of the At2S2 promoter. Thus, the BRM-containing chromatin-remodeling ATPase complex involved in many aspects of plant development mediates the repression of SSPs in leaf tissue. PMID- 18508956 TI - Magnitude and direction of vesicle dynamics in growing pollen tubes using spatiotemporal image correlation spectroscopy and fluorescence recovery after photobleaching. AB - The delivery of cell wall material and membrane to growing plant cell surfaces requires the spatial and temporal coordination of secretory vesicle trafficking. Given the small size of vesicles, their dynamics is difficult to quantify. To quantitatively analyze vesicle dynamics in growing pollen tubes labeled with the styryl dye FM1-43, we applied spatiotemporal correlation spectroscopy on time lapse series obtained with high-speed confocal laser scanning microscopy recordings. The resulting vector maps revealed that vesicles migrate toward the apex in the cell cortex and that they accumulate in an annulus-shaped region adjacent to the extreme tip and then turn back to flow rearward in the center of the tube. Fluorescence recovery after photobleaching confirmed vesicle accumulation in the shoulder of the apex, and it revealed that the extreme apex never recovers full fluorescence intensity. This is consistent with endocytotic activity occurring in this region. Fluorescence recovery after photobleaching analysis also allowed us to measure the turnover rate of the apical vesicle population, which was significantly more rapid than the theoretical rate computed based on requirements for new cell wall material. This may indicate that a significant portion of the vesicles delivered to the apex does not succeed in contacting the plasma membrane for delivery of their contents. Therefore, we propose that more than one passage into the apex may be needed for many vesicles before they fuse to the plasma membrane and deliver their contents. PMID- 18508957 TI - SCAMPs highlight the developing cell plate during cytokinesis in tobacco BY-2 cells. AB - We previously demonstrated that rice (Oryza sativa) SECRETORY CARRIER MEMBRANE PROTEIN1 (OsSCAMP1)-yellow fluorescent protein in transgenic tobacco (Nicotiana tabacum) Bright Yellow-2 cells locates to the plasma membrane and to motile punctate structures, which represent the trans-Golgi network/early endosome and are tubular-vesicular in nature. Here, we now show that SCAMPs are diverted to the cell plate during cytokinesis dividing Bright Yellow-2 cells. As cells progress from metaphase to cytokinesis, punctate OsSCAMP1-labeled structures begin to collect in the future division plane. Together with the internalized endosomal marker FM4-64, they then become incorporated into the cell plate as it forms and expands. This was confirmed by immunogold electron microscopy. We also monitored for the Golgi apparatus and the prevacuolar compartment (PVC)/multivesicular body. Golgi stacks tend to accumulate in the vicinity of the division plane, but the signals are clearly separate to the cell plate. The situation with the PVC (labeled by green fluorescent protein-BP-80) is not so clear. Punctate BP-80 signals are seen at the advancing periphery of the cell plate, which was confirmed by immunogold electron microscopy. Specific but weak labeling was observed in the cell plate, but no evidence for a fusion of the PVC/multivesicular body with the cell plate could be obtained. Our data, therefore, support the notion that cell plate formation is mainly a secretory process involving mass incorporation of domains of the trans-Golgi network/early endosome membrane. We regard the involvement of multivesicular late endosomes in this process to be equivocal. PMID- 18508958 TI - Physiological and transcriptomic aspects of urea uptake and assimilation in Arabidopsis plants. AB - Urea is the major nitrogen (N) form supplied as fertilizer in agriculture, but it is also an important N metabolite in plants. Urea transport and assimilation were investigated in Arabidopsis (Arabidopsis thaliana). Uptake studies using (15)N labeled urea demonstrated the capacity of Arabidopsis to absorb urea and that the urea uptake was regulated by the initial N status of the plants. Urea uptake was stimulated by urea but was reduced by the presence of ammonium nitrate in the growth medium. N deficiency in plants did not affect urea uptake. Urea exerted a repressive effect on nitrate influx, whereas urea enhanced ammonium uptake. The use of [(15)N]urea and [(15)N]ammonium tracers allowed us to show that urea and ammonium assimilation pathways were similar. Finally, urea uptake was less efficient than nitrate uptake, and urea grown-plants presented signs of N starvation. We also report the first analysis, to our knowledge, of Arabidopsis gene expression profiling in response to urea. Our transcriptomic approach revealed that nitrate and ammonium transporters were transcriptionally regulated by urea as well as key enzymes of the glutamine synthetase-glutamate synthase pathway. AtDUR3, a high-affinity urea transporter in Arabidopsis, was strongly up regulated by urea. Moreover, our transcriptomic data suggest that other genes are also involved in urea influx. PMID- 18508959 TI - Sequence analysis of bacterial artificial chromosome clones from the apospory specific genomic region of Pennisetum and Cenchrus. AB - Apomixis, asexual reproduction through seed, is widespread among angiosperm families. Gametophytic apomixis in Pennisetum squamulatum and Cenchrus ciliaris is controlled by the apospory-specific genomic region (ASGR), which is highly conserved and macrosyntenic between these species. Thirty-two ASGR bacterial artificial chromosomes (BACs) isolated from both species and one ASGR-recombining BAC from P. squamulatum, which together cover approximately 2.7 Mb of DNA, were used to investigate the genomic structure of this region. Phrap assembly of 4,521 high-quality reads generated 1,341 contiguous sequences (contigs; 730 from the ASGR and 30 from the ASGR-recombining BAC in P. squamulatum, plus 580 from the C. ciliaris ASGR). Contigs containing putative protein-coding regions unrelated to transposable elements were identified based on protein similarity after Basic Local Alignment Search Tool X analysis. These putative coding regions were further analyzed in silico with reference to the rice (Oryza sativa) and sorghum (Sorghum bicolor) genomes using the resources at Gramene (www.gramene.org) and Phytozome (www.phytozome.net) and by hybridization against sorghum BAC filters. The ASGR sequences reveal that the ASGR (1) contains both gene-rich and gene-poor segments, (2) contains several genes that may play a role in apomictic development, (3) has many classes of transposable elements, and (4) does not exhibit large-scale synteny with either rice or sorghum genomes but does contain multiple regions of microsynteny with these species. PMID- 18508960 TI - Glucocorticoid-mediated hypertension: does the vascular smooth muscle hold all the answers? PMID- 18508961 TI - Marconi revisited: from kidney to brain--two organ systems communicating at long distance. PMID- 18508963 TI - Resolved: Fistulas are preferred to grafts as initial vascular access for dialysis. Pro. AB - There is growing concern that the Fistula First Initiative, KDOQI guidelines, and subsequent pressure from the Centers for Medicare and Medicaid Services lack reasonableness regarding likely success for fistula maturation in a heterogeneous, new-onset dialysis population. Here the various positions are examined from multiple perspectives. PMID- 18508962 TI - Organic anion transporter 3 contributes to the regulation of blood pressure. AB - Renal organic anion transporters (OAT) are known to mediate the excretion of many drugs, but their function in normal physiology is not well understood. In this study, mice lacking organic anion transporter 3 (Oat3) had a 10 to 15% lower BP than wild-type mice, raising the possibility that Oat3 transports an endogenous regulator of BP. The aldosterone response to a low-salt diet was blunted in Oat3 null mice, but baseline aldosterone concentration was higher in these mice, suggesting that aldosterone dysregulation does not fully explain the lower BP in the basal state; therefore, both targeted and global metabolomic analyses of plasma and urine were performed, and several potential endogenous substrates of Oat3 were found to accumulate in the plasma of Oat3-null mice. One of these substrates, thymidine, was transported by Oat3 expressed in vitro. In vivo, thymidine, as well as two of the most potent Oat3 inhibitors that were characterized, reduced BP by 10 to 15%; therefore, Oat3 seems to regulate BP, and Oat3 inhibitors might be therapeutically useful antihypertensive agents. Moreover, polymorphisms in human OAT3 might contribute to the genetic variation in susceptibility to hypertension. PMID- 18508964 TI - Fructose induces the inflammatory molecule ICAM-1 in endothelial cells. AB - Epidemiologic studies have linked fructose intake with the metabolic syndrome, and it was recently reported that fructose induces an inflammatory response in the rat kidney. Here, we examined whether fructose directly stimulates endothelial inflammatory processes by upregulating the inflammatory molecule intercellular adhesion molecule-1 (ICAM-1). When human aortic endothelial cells were stimulated with physiologic concentrations of fructose, ICAM-1 mRNA and protein expression increased in a time- and dosage-dependent manner, which was independent of NF-kappaB activation. Fructose reduced endothelial nitric oxide (NO) levels and caused a transient reduction in endothelial NO synthase expression. The administration of an NO donor inhibited fructose-induced ICAM-1 expression, whereas blocking NO synthase enhanced it, suggesting that NO inhibits endothelial ICAM-1 expression. Furthermore, fructose resulted in decreased intracellular ATP; administration of exogenous ATP blocked fructose-induced ICAM 1 expression and increased NO levels. Consistent with the in vitro studies, dietary intake of fructose at physiologic dosages increased both serum ICAM-1 concentration and endothelial ICAM-1 expression in the rat kidney. These data suggest that fructose induces inflammatory changes in vascular cells at physiologic concentrations. PMID- 18508965 TI - Association of incident gout and mortality in dialysis patients. AB - Previous studies have shown that gout is associated with an increased risk for cardiovascular mortality in the general population, but this has not been well studied in patients with ESRD. In this study, the incidence of gout and its association with mortality was evaluated in 259,209 patients in the United States Renal Data System. Overall, the incidence of gout in the first year of dialysis was 5% and in the first 5 yr was 15.4%. Independent risk factors for gout in adjusted analyses included black race, older age, female gender, hypertension, ischemic heart disease, congestive heart failure, and alcohol use. Factors associated with a lower risk for gout included a history of diabetes, smoking, and peripheral vascular disease. Time-dependent Cox regression analysis suggested that an episode of gout was independently associated with a 1.5-fold increase in mortality risk (adjusted hazard ratio 1.49; 95% confidence interval 1.43 to 1.55). The mechanisms underlying this association require further study. PMID- 18508966 TI - Characterization of the sodium/hydrogen exchanger NHA2. AB - Cation/proton exchange has been recognized for decades in mammalian mitochondria, but the exchanger proteins have eluded identification. In this study, a cDNA from a human brain library, previously designated NHA2 in the genome, was cloned and characterized. The NHA2 transcript bears more similarity to prokaryotic than known eukaryotic sodium/proton exchangers, but it was found to be expressed in multiple mammalian organs and cultured cells. A mAb to NHA2 was generated and found to label an approximately 55-kD native protein in multiple tissues and cell lines. The specificity of this antibody was confirmed by demonstrating the loss of the native NHA2 band on immunoblots when cultured cells were treated with NHA2 specific small interfering RNA. Although NHA2 protein was detected in multiple organs, within each, its expression was restricted to specific cell types. In the kidney, co-localization with calbindin 28k and reverse transcription-PCR of microdissected tubules revealed that NHA2 is limited to the distal convoluted tubule. In cell lines, native NHA2 was localized both to the plasma membrane and to the intracellular compartment; immunogold electron microscopy of rat distal convoluted tubule demonstrated NHA2 predominantly but not exclusively on the inner mitochondrial membrane. Furthermore, co-sedimentation of NHA2 antigen and mitochondrial membranes was observed with differential centrifugation, and two mitochondrial markers co-localized with NHA2 in cultured cells. Regarding function, human NHA2 reversed the sodium/hydrogen exchanger-null phenotype when expressed in sodium/hydrogen exchanger-deficient yeast and restored the ability to defend high salinity in the presence of acidic extracellular pH. In summary, NHA2 is a ubiquitous mammalian sodium proton/exchanger that is restricted to the distal convoluted tubule in the kidney. PMID- 18508968 TI - The shortest wire. PMID- 18508967 TI - IHG-1 amplifies TGF-beta1 signaling and is increased in renal fibrosis. AB - Induced in high glucose-1 (IHG-1) is an evolutionarily conserved gene transcript upregulated by high extracellular glucose concentrations, but its function is unknown. Here, it is reported that the abundance of IHG-1 mRNA is nearly 10-fold higher in microdissected, tubule-rich renal biopsies from patients with diabetic nephropathy compared with control subjects. In the diabetic nephropathy specimens, in situ hybridization localized IHG-1 to tubular epithelial cells along with TGF-beta1 and activated Smad3, suggesting a possible role in the development of tubulointerstitial fibrosis. Supporting this possibility, IHG-1 mRNA and protein expression also increased with unilateral ureteral obstruction. In the HK-2 proximal tubule cell line, overexpression of IHG-1 increased TGF beta1-stimulated expression of connective tissue growth factor and fibronectin. IHG-1 was found to amplify TGF-beta1-mediated transcriptional activity by increasing and prolonging phosphorylation of Smad3. Conversely, inhibition of endogenous IHG-1 with small interference RNA suppressed transcriptional responses to TGF-beta1. In summary, IHG-1, which increases in diabetic nephropathy, may enhance the actions of TGF-beta1 and contribute to the development of tubulointerstitial fibrosis. PMID- 18508969 TI - Synthesis of linked carbon monolayers: films, balloons, tubes, and pleated sheets. AB - Because of their potential for use in advanced electronic, nanomechanical, and other applications, large two-dimensional, carbon-rich networks have become an important target to the scientific community. Current methods for the synthesis of these materials have many limitations including lack of molecular-level control and poor diversity. Here, we present a method for the synthesis of two dimensional carbon nanomaterials synthesized by Mo- and Cu-catalyzed cross linking of alkyne-containing self-assembled monolayers on SiO(2) and Si(3)N(4). When deposited and cross-linked on flat surfaces, spheres, cylinders, or textured substrates, monolayers take the form of these templates and retain their structure on template removal. These nanomaterials can also be transferred from surface to surface and suspended over cavities without tearing. This approach to the synthesis of monolayer carbon networks greatly expands the chemistry, morphology, and size of carbon films accessible for analysis and device applications. PMID- 18508970 TI - Synthesis and bioassay of improved mosquito repellents predicted from chemical structure. AB - Mosquito repellency data on acylpiperidines derived from the U.S. Department of Agriculture archives were modeled by using molecular descriptors calculated by CODESSA PRO software. An artificial neural network model was developed for the correlation of these archival results and used to predict the repellent activity of novel compounds of similar structures. A series of 34 promising N acylpiperidine mosquito repellent candidates (4a-4q') were synthesized by reactions of acylbenzotriazoles 2a-2p with piperidines 3a-3f. Compounds (4a-4q') were screened as topically applied mosquito repellents by measuring the duration of repellency after application to cloth patches worn on the arms of human volunteers. Some compounds that were evaluated repelled mosquitoes as much as three times longer than N,N-diethyl-m-toluamide (DEET), the most widely used repellent throughout the world. The newly measured durations of repellency were used to obtain a superior correlation equation relating mosquito repellency to molecular structure. PMID- 18508971 TI - The evolution of courtship behaviors through the origination of a new gene in Drosophila. AB - New genes can originate by the combination of sequences from unrelated genes or their duplicates to form a chimeric structure. These chimeric genes often evolve rapidly, suggesting that they undergo adaptive evolution and may therefore be involved in novel phenotypes. Their functions, however, are rarely known. Here, we describe the phenotypic effects of a chimeric gene, sphinx, that has recently evolved in Drosophila melanogaster. We show that a knockout of this gene leads to increased male-male courtship in D. melanogaster, although it leaves other aspects of mating behavior unchanged. Comparative studies of courtship behavior in other closely related Drosophila species suggest that this mutant phenotype of male-male courtship is the ancestral condition because these related species show much higher levels of male-male courtship than D. melanogaster. D. melanogaster therefore seems to have evolved in its courtship behaviors by the recruitment of a new chimeric gene. PMID- 18508972 TI - The evolution of imperfect floral mimicry. AB - The theory of mimicry predicts that selection favors signal refinement in mimics to optimally match the signals released by their specific model species. We provide here chemical and behavioral evidence that a sexually deceptive orchid benefits from its mimetic imperfection to its co-occurring and specific bee model by triggering a stronger response in male bees, which react more intensively to the similar, but novel, scent stimulus provided by the orchid. PMID- 18508973 TI - Social interaction-mediated lifespan extension of Drosophila Cu/Zn superoxide dismutase mutants. AB - Beneficial effects of social interaction on aging have been studied in humans and other species. We found that short-lived Drosophila mutants of the antioxidant enzyme Cu/Zn superoxide dismutase displayed a robust lifespan extension, with improved stress resistance and motor ability, upon cohousing with active flies of longer lifespan or younger age. Genetic, surgical, and environmental manipulations revealed motor and sensory components in behavioral interactions required for the lifespan extension induced by cohousing. Our results provide a definitive case of beneficial social interaction on lifespan and a useful entry point for analyzing the underlying molecular networks and physiological mechanisms. PMID- 18508974 TI - Chronic sodium nitrite therapy augments ischemia-induced angiogenesis and arteriogenesis. AB - Chronic tissue ischemia due to defective vascular perfusion is a hallmark feature of peripheral artery disease for which minimal therapeutic options exist. We have reported that sodium nitrite therapy exerts cytoprotective effects against acute ischemia/reperfusion injury in both heart and liver, consistent with the model of bioactive NO formation from nitrite during ischemic stress. Here, we test the hypothesis that chronic sodium nitrite therapy can selectively augment angiogenic activity and tissue perfusion in the murine hind-limb ischemia model. Various therapeutic doses (8.25-3,300 mug/kg) of sodium nitrite or PBS were administered. Sodium nitrite significantly restored ischemic hind-limb blood flow in a time dependent manner, with low-dose sodium nitrite being most effective. Nitrite therapy significantly increased ischemic limb vascular density and stimulated endothelial cell proliferation. Remarkably, the effects of sodium nitrite therapy were evident within 3 days of the ischemic insult demonstrating the potency and efficacy of chronic sodium nitrite therapy. Sodium nitrite therapy also increased ischemic tissue nitrite and NO metabolites compared to nonischemic limbs. Use of the NO scavenger carboxy PTIO completely abolished sodium nitrite-dependent ischemic tissue blood flow and angiogenic activity consistent with nitrite reduction to NO being the proangiogenic mechanism. These data demonstrate that chronic sodium nitrite therapy is a recently discovered therapeutic treatment for peripheral artery disease and critical limb ischemia. PMID- 18508975 TI - Contemporary North American influenza H7 viruses possess human receptor specificity: Implications for virus transmissibility. AB - Avian H7 influenza viruses from both the Eurasian and North American lineage have caused outbreaks in poultry since 2002, with confirmed human infection occurring during outbreaks in The Netherlands, British Columbia, and the United Kingdom. The majority of H7 infections have resulted in self-limiting conjunctivitis, whereas probable human-to-human transmission has been rare. Here, we used glycan microarray technology to determine the receptor-binding preference of Eurasian and North American lineage H7 influenza viruses and their transmissibility in the ferret model. We found that highly pathogenic H7N7 viruses from The Netherlands in 2003 maintained the classic avian-binding preference for alpha2-3-linked sialic acids (SA) and are not readily transmissible in ferrets, as observed previously for highly pathogenic H5N1 viruses. However, H7N3 viruses isolated from Canada in 2004 and H7N2 viruses from the northeastern United States isolated in 2002-2003 possessed an HA with increased affinity toward alpha2-6-linked SA, the linkage type found prominently on human tracheal epithelial cells. We identified a low pathogenic H7N2 virus isolated from a man in New York in 2003, A/NY/107/03, which replicated efficiently in the upper respiratory tract of ferrets and was capable of transmission in this species by direct contact. These results indicate that H7 influenza viruses from the North American lineage have acquired sialic acid-binding properties that more closely resemble those of human influenza viruses and have the potential to spread to naive animals. PMID- 18508976 TI - In vivo two-photon voltage-sensitive dye imaging reveals top-down control of cortical layers 1 and 2 during wakefulness. AB - Conventional methods of imaging membrane potential changes have limited spatial resolution, particularly along the axis perpendicular to the cortical surface. The laminar organization of the cortex suggests, however, that the distribution of activity in depth is not uniform. We developed a technique to resolve network activity of different cortical layers in vivo using two-photon microscopy of the voltage-sensitive dye (VSD) ANNINE-6. We imaged spontaneous voltage changes in the barrel field of the somatosensory cortex of head-restrained mice and analyzed their spatiotemporal correlations during anesthesia and wakefulness. EEG recordings always correlated more strongly with VSD signals in layer (L) 2 than in L1. Nearby (<200 mum) cortical areas were correlated with one another during anesthesia. Waking the mouse strongly desynchronized neighboring cortical areas in L1 in the 4- to 10-Hz frequency band. Wakefulness also slightly increased synchrony of neighboring territories in L2 in the 0.5- to 4.0-Hz range. Our observations are consistent with the idea that, in the awake animal, long-range inputs to L1 of the sensory cortex from various cortical and thalamic areas exert top-down control on sensory processing. PMID- 18508977 TI - Outcome of single-bundle versus double-bundle reconstruction of the anterior cruciate ligament: a meta-analysis. AB - BACKGROUND: The anterior cruciate ligament (ACL) has 2 anatomic bundles. Standard ACL reconstruction is with a single-bundle graft, but double-bundle reconstruction may better control knee rotational torque, a potential cause of failure after single-bundle reconstruction. The authors investigated outcomes of single-bundle versus double-bundle ACL reconstruction. HYPOTHESIS: There is no difference in outcomes of single-bundle versus double-bundle reconstruction. STUDY DESIGN: Meta-analysis. METHODS: The authors systematically identified randomized controlled trials (RCTs) comparing single-bundle versus double-bundle ACL reconstruction (secondary analysis includes nonrandomized trials). Outcomes reported in a majority of included trials were meta-analyzed. RESULTS: Four RCTs were included (secondary analysis including 5 additional trials yielded reassuringly similar results). Two outcome measures were reported (in a manner permitting meta-analysis) in at least 3 of 4 trials: KT-1000 arthrometer and pivot-shift testing. On average, KT-1000 arthrometer side-to-side difference was 0.52 mm closer to normal in patients treated with double-bundle reconstruction. This difference is demonstrated to be clinically insignificant. In addition, there was no statistical difference in the odds of having a normal or nearly normal pivot-shift result in patients treated with double-bundle versus single bundle reconstruction. CONCLUSION: Double-bundle reconstruction does not result in clinically significant differences in KT-1000 arthrometer or pivot-shift testing. The pivot-shift results have particular clinical relevance because the test is designed to evaluate knee rotational instability; the results do not support the theory that double-bundle reconstruction better controls knee rotation. Improved quality of future RCTs would allow meta-analysis of a greater number of outcome measures including measures of symptoms and disabilities most important to patients. PMID- 18508978 TI - Scaling up antiretroviral therapy in developing countries: what are the benefits and challenges? PMID- 18508979 TI - Chemoprevention and screening in primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic inflammatory disease that causes fibrosis of the biliary tree. Life expectancy of patients is reduced by liver failure and a high incidence of malignancy. It is closely associated with inflammatory bowel disease, particularly ulcerative colitis, which coexists in approximately three-quarters of northern European patients. Cancers include cholangiocarcinoma, gallbladder cancer, hepatocellular carcinoma, pancreatic cancer and colorectal cancer. Ursodeoxycholic acid appears to reduce the incidence of colorectal neoplasia in patients with PSC, and there is some suggestion that it may also reduce the incidence of cholangiocarcinoma. A chemoprotective benefit of 5-aminosalicylates has not been confirmed in patients with PSC with associated inflammatory bowel disease. There is no accepted screening programme for cholangiocarcinoma, but methods for detecting early disease using biochemical markers, scanning using positron emission tomography or MRI, and endoscopic procedures such as endosonography and endoscopic retrograde cholangiopancreatography are discussed. A combination of techniques is often used in an attempt to diagnose early cholangiocarcinoma. Cholecystectomy should be performed for gallbladder polyps, as many are malignant, and ultrasonography and alpha-fetoprotein testing are suggested for screening for hepatocellular carcinoma. Colorectal carcinoma screening should be performed after the diagnosis of PSC, and surveillance colonoscopy should be performed annually if there is concomitant colitis. PMID- 18508980 TI - Stroke in pregnancy and the puerperium. AB - Stroke is a recognised complication of pregnancy, contributing to more than 12% of all maternal deaths. Estimated incidence rates vary considerably from 4.3 to 210 strokes per 100,000 deliveries. Atherosclerosis is rare in young adults, and so other causes of stroke become increasingly likely. Aetiological factors important in pregnancy include hypercoagulability due to maternal physiological changes, pre-eclampsia and eclampsia, cerebral venous thrombosis, paradoxical embolism, postpartum cerebral angiopathy and peripartum cardiomyopathy. Management of patients with pregnancy-related stroke should generally proceed as for non-pregnant patients, although there are a number of important areas specific to pregnancy which will be considered here. PMID- 18508981 TI - Clinical applications and advances of positron emission tomography with fluorine 18-fluorodeoxyglucose (18F-FDG) in the diagnosis of liver neoplasms. AB - Positron emission tomography (PET) using 18F-fluorodeoxyglucose (18F-FDG) has been widely used not only for detecting and staging malignant tumours but also for monitoring therapy response and for differentiating malignant lesions from benign lesions. However, the sensitivity of 18F-FDG PET for hepatocellular carcinoma (HCC) is low. Recent research about the novel drug shows that the sensitivity of 11C-acetate PET for HCC is higher than that of 18F-FDG PET and 11C acetate also indicates tumour cell differentiation. 18F-FDG PET proved to be highly sensitive in detecting hepatic metastases from different primaries. 18F FDG PET has also been shown to be useful and more accurate than conventional imaging modalities in demonstrating the effect of local treatments in liver tumours. With the development of small animal imaging, animal research about 18F FDG PET is increasing. PMID- 18508982 TI - Antithrombotic treatment in atrial fibrillation. AB - Atrial fibrillation (AF) can significantly increase morbidity and mortality. It is gaining in clinical and economic importance, being the most commonly encountered tachyarrhythmia in clinical practice. Stroke is the most serious complication. Evidence from AF antithrombotic treatment trials is reviewed, risk stratification of patients with AF is discussed, and recommendations for anticoagulation are presented. PMID- 18508983 TI - Pathological networking: a new approach to understanding COPD. AB - Developing new treatments for chronic obstructive pulmonary disease (COPD) is extremely challenging. This disease, chronic by definition, becomes apparent only after substantial--and probably irreversible--tissue damage has occurred. The observable phenotype is of a stable disease state whose progression is hard to influence and reversal of which appears almost impossible. Identifying key components of the pathological process, targeting of which will result in substantial clinical benefit, is a significant challenge. In this review the nature of the disease is examined and conceptual information and simple tissue models of inflammation are used to explore the pathological network that is COPD. From the concept of COPD as a disease network displaying the features of contiguous immunity (in which many processes of innate and adaptive immunity are in continual dialogue and evolution), refinements are suggested to the strategies aimed at developing effective new treatments for this disease. PMID- 18508984 TI - Management of septic arthritis: a systematic review. AB - OBJECTIVE: To evaluate the existing evidence on the diagnosis and management of septic arthritis in native joints. DESIGN: Systematic review. DATA SOURCES: Cochrane Library, Medline, Embase, National Electronic Library for Health, reference lists, national experts. REVIEW METHODS: Systematic review of the literature with evaluation of the methodological quality of the selected papers using defined criteria set out by the Clinical Effectiveness and Evaluation Unit of the Royal College of Physicians. RESULTS: 3291 citations were initially identified. Of these, 189 full text articles were identified for potential selection. Following review of these full text articles, 80 articles were found to fulfil the inclusion criteria and were included in the final list. Conclusions were drawn on the diagnosis, investigation and management of septic arthritis. DISCUSSION: Little good quality evidence exists to guide the diagnosis and management of septic arthritis. Overall, no investigation is more reliable in the diagnosis of septic arthritis than the opinion of an experienced doctor. Aspiration and culture of synovial fluid is crucial to the diagnosis, but measurement of cell count is unhelpful. Antibiotics are clearly required for a prolonged period, but there are no data to indicate by which route or for how long. Key unanswered questions remain surrounding the medical and surgical management of the infected joint. PMID- 18508985 TI - Progression of early warning scores (EWS) in patients with acute pancreatitis: a re-evaluation of a retrospective cohort of patients. AB - INTRODUCTION: Early warning scores (EWS) is a physiological scoring system measured hourly. This study determined how progression of EWS affected outcome in acute pancreatitis. METHODS: The single worst EWS score for each 24 h period following admission was recorded for 110 patients with acute pancreatitis. Scores falling below 3 were defined as improving; scores which remained at 3 or rising were considered deteriorating. RESULTS: Deteriorating EWS values were associated with a greatly increased risk of mortality (p<0.001). All patients within the groups, who died, had an adverse outcome or had severe pancreatitis, demonstrated a failure to improve on a median admission EWS of >or=3 or a deterioration of their median EWS to above this. This progression occurred within 48 h of admission. Evaluating the progression of EWS (that is, improving or deteriorating scores) resulted in an improvement in the sensitivity and specificity in predicting adverse outcome, mortality or severe pancreatitis when compared to previously published data on EWS scores alone, on days 1 to 3 following admission. CONCLUSION: Deteriorating EWS values within the 48 h from admission are associated with adverse outcome or death in acute pancreatitis. Measuring progression of EWS over 72 h from admission can further improve accuracy of this monitoring system for acute pancreatitis. PMID- 18508986 TI - Non-Hodgkin's lymphoma and tuberculosis coexistence in the same organs: a report of two cases. AB - Non-Hodgkin's lymphoma (NHL) may be preceded by chronic inflammatory diseases and furthermore has been related to immune deficiency. Tuberculosis (TB), on the other hand, is a chronic infectious disease whose presentation and reactivation is known to be promoted by cell mediated immunodeficiency. The coexistence of NHL and TB in the same organ is rare. We report two cases of NHL and TB coexistence in two different organs: cervical lymph nodes and kidney. The cases illustrate how misleading the concurrence of NHL and TB infection can be, delaying the diagnosis and treatment of either disease. PMID- 18508987 TI - Resident's knowledge on thoracic aortic dissection. PMID- 18508988 TI - The facts of death. PMID- 18508989 TI - How much infertility does chlamydia cause? PMID- 18508990 TI - Two heads are better than one: the association between condom decision-making and condom use errors and problems. AB - OBJECTIVES: This exploratory study compared the frequency of condom use errors and problems between men reporting that condom use for penile-vaginal sex was a mutual decision compared with men making the decision unilaterally. METHODS: Nearly 2000 people completed a web-based questionnaire. A sub-sample of 660 men reporting that they last used a condom for penile-vaginal sex (within the past three months) was analysed. Nine condom use errors/problems were assessed. Multivariate analyses controlled for men's age, marital status, and level of experience using condoms. RESULTS: Men's unilateral decision-making was associated with increased odds of removing condoms before sex ended (adjusted odds ratio (AOR) 2.51, p = 0.002), breakage (AOR 3.90, p = 0.037), and slippage during withdrawal (AOR 2.04, p = 0.019). Men's self-reported level of experience using condoms was significantly associated with seven out of nine errors/problems, with those indicating less experience consistently reporting more errors/problems. CONCLUSIONS: Findings suggest that female involvement in the decision to use condoms for penile-vaginal sex may be partly protective against some condom errors/problems. Men's self-reported level of experience using condoms may be a useful indicator of the need for education designed to promote the correct use of condoms. Education programmes may benefit men by urging them to involve their female partner in condom use decisions. PMID- 18508995 TI - Endocannabinoids in endocrine and related tumours. AB - The 'endocannabinoid system', comprising the cannabinoid CB1 and CB2 receptors, their endogenous ligands, endocannabinoids and the enzymes that regulate their biosynthesis and degradation, has drawn a great deal of scientist attention during the last two decades. The endocannabinoid system is involved in a broad range of functions and in a growing number of physiopathological conditions. Indeed, recent evidence indicates that endocannabinoids influence the intracellular events controlling the proliferation of numerous types of endocrine and related cancer cells, thereby leading to both in vitro and in vivo antitumour effects. In particular, they are able to inhibit cell growth, invasion and metastasis of thyroid, breast and prostate tumours. The chief events of endocannabinoids in cancer cell proliferation are reported highlighting the correspondent signalling involved in tumour processes: regulation of adenylyl cyclase, cyclic AMP-protein kinase-A pathway and MEK-extracellular signal regulated kinase signalling cascade. PMID- 18508994 TI - Signal transducer and activator of transcription 5A/B in prostate and breast cancers. AB - Protein kinase signaling pathways, such as Janus kinase 2-Signal transducer and activator of transcription 5A/B (JAK2-STAT5A/B), are of significant interest in the search for new therapeutic strategies in both breast and prostate cancers. In prostate cancer, the components of the JAK2-STAT5A/B signaling pathway provide molecular targets for small-molecule inhibition of survival and growth signals of the cells. At the same time, new evidence suggests that the STAT5A/B signaling pathway is involved in the transition of organ-confined prostate cancer to hormone-refractory disease. This implies that the active JAK2-STAT5A/B signaling pathway potentially provides the means for pharmacological intervention of clinical prostate cancer progression. In addition, active STAT5A/B may serve as a prognostic marker for identification of those primary prostate cancers that are likely to progress to aggressive disease. In breast cancer, the role of STAT5A/B is more complex. STAT5A/B may have a dual role in the regulation of malignant mammary epithelium. Data accumulated from mouse models of breast cancer suggest that in early stages of breast cancer STAT5A/B may promote malignant transformation and enhance growth of the tumor. This is in contrast to established breast cancer, where STAT5A/B may mediate the critical cues for maintaining the differentiation of mammary epithelium. In addition, present data suggest that activation of STAT5A/B in breast cancer predicts favorable clinical outcome. The dual nature of STAT5A/B action in breast cancer makes the therapeutic use of STAT5 A/B more complex. PMID- 18508997 TI - Patterns and changes in gene expression following neo-adjuvant anti-estrogen treatment in estrogen receptor-positive breast cancer. AB - This study aimed to define a gene expression profile associated with response to anti-estrogen treatment in estrogen receptor alpha (ERalpha)-positive breast cancer from elderly patients and to identify possible candidate genes associated with resistance by detecting those modulated by treatment. Using cDNA microarrays containing 16 702 unique clones, 21 pre-treatment and 11 paired post-treatment samples collected in a neo-adjuvant toremifene trial on elderly patients with operable and locally advanced ERalpha-positive breast cancer were profiled. Gene expression profiles generated from pre-treatment samples were correlated with treatment-induced tumor shrinkage and compared with those obtained from post treatment paired samples to define genes differentially modulated following anti estrogen treatment. Correlation analysis on 21 pre-treatment samples highlighted 53 genes significantly related to treatment response (P<0.001). Genes involved in cell cycle and proliferation were more frequently upregulated in responders compared with non-responders. Class comparison analysis identified 101 genes significantly modulated independently of treatment response; 82 genes were modulated in non-responders, whereas only 8 genes were differently expressed after treatment in responders. Gene expression profiles appear to be more frequently modulated by anti-estrogen treatment in non-responding patients and may harbor interesting genes possibly involved in anti-estrogen resistance, including clusterin, MAPK6, and MMP2. This concept was corroborated by in vitro studies showing that silencing of CLU restored toremifene sensitivity in the ER anti-estrogen-resistant breast cancer cell line T47D. Integration between neo adjuvant therapy and transcriptional profiling has therefore the potential to identify therapeutic targets to be challenged for overcoming treatment resistance. PMID- 18508998 TI - Estrogen signaling ability in human endometrial cancer through the cancer-stromal interaction. AB - The estrogen pathway plays an important role in the etiology of human endometrial carcinoma (EC). We examined whether estrogen biosynthesis in the tumor microenvironment promotes endometrial cancer. To examine the contribution of stromal cells to estrogen signaling in EC, we used reporter cells stably transfected with the estrogen response element (ERE) fused to the destabilized green fluorescent protein (GFP) gene. In this system, the endometrial cancer stromal cells from several patients activated the ERE of cancer cells to a variable extent. The GFP expression level increased when testosterone, a substrate for aromatase, was added. The effect was variably inhibited by aromatase inhibitors (AIs), although the response to AIs varied among patients. These results suggest that GFP expression is driven by estrogen synthesized by aromatase in the endometrial cancer stromal cells. In a second experiment, we constructed an adenovirus reporter vector containing the same construct as the reporter cells described above, and visualized endogenous ERE activity in primary culture cancer cells from 15 EC specimens. The GFP expression levels varied among the cases, and in most primary tissues, ERE activities were strongly inhibited by a pure anti-estrogen, fulvestrant. Interestingly, a minority of primary tissues in endometrial cancer showed ERE activity independent of the estrogen-ER pathway. These results suggest that AI may have some therapeutic value in EC; however, the hormonal microenvironment must be assessed prior to initiating therapy. PMID- 18508996 TI - Pancreatic endocrine neoplasms: epidemiology and prognosis of pancreatic endocrine tumors. AB - Pancreatic endocrine tumors (PETs) are uncommon tumors with an annual incidence <1 per 100 000 person-years in the general population. The PETs that produce hormones resulting in symptoms are designated as functional. The majority of PETs are non-functional. Of the functional tumors, insulinomas are the most common, followed by gastrinomas. The clinical course of patients with PETs is variable and depends on the extent of the disease and the treatment rendered. Patients with completely resected tumors generally have a good prognosis, and aggressive surgical therapy in patients with advanced disease may also prolong survival. The epidemiology, prognosis, and established and novel prognostic markers of PETs are reviewed. PMID- 18508999 TI - Expression of progesterone and estradiol receptors in normal adrenal cortex, adrenocortical tumors, and primary pigmented nodular adrenocortical disease. AB - Adrenal tumors occur more frequently in women and are the leading cause of Cushing's syndrome during pregnancy. We aimed to evaluate the potential role of sex steroids in the susceptibility of women to adrenocortical tumors. We evaluated the presence of the progesterone receptor (PR), estradiol receptors (ERs), and aromatase in 5 patients with primary pigmented nodular adrenal disease (PPNAD), 15 adrenocortical adenomas (ACAs) and adjacent normal tissues, 12 adrenocortical carcinomas (ACCs), and 3 normal adrenal glands (NA). The expression of PR and ERalpha was evaluated by enzyme immunoassays, real-time RT PCR, immunohistochemistry, and cytosol-based ligand-binding assays. ERbeta and aromatase levels were evaluated by real-time RT-PCR. ERalpha concentrations were low in NA, in adrenal tissues adjacent to ACA (51+/-33), in ACC (53+/-78), and lower in ACA (11+/-11 fmol/mg DNA). Conversely, PR concentrations were high in NA and adrenal tissues adjacent to ACA, at 307+/-216 fmol/mg DNA, and were even higher in tumors - 726+/-706 fmol/mg DNA in ACA and 1154+/-1586 fmol/mg DNA in ACC - and in isolated PPNAD nodules. Binding study results in four tumors were compatible with binding to a steroid receptor. In patients with PPNAD, a strong positive immunohistochemical signal was associated with the sole isolated nodular regions. ERbeta transcript levels were very high in all samples except those for two ACCs, whereas aromatase levels were low. PR and ERbeta are clearly present in normal adrenal glands and adrenal tumors. Further studies may shed light on the possible pathogenic role of these receptors in adrenal proliferation. PMID- 18509000 TI - Estrogen promotes tumor progression in a genetically defined mouse model of lung adenocarcinoma. AB - Numerous epidemiological observations point to sex differences in lung cancer etiology and progression. The present study was aimed at understanding the bases of these sex differences. To test the effect of estradiol on tumor progression, we used a mouse model based on conditional Kras expression and concurrent deletion of Tp53 following inhalation of an adenoviral vector expressing Cre recombinase (AdeCre). Ovariectomized females and males were treated with estradiol via a continuous-release capsule. Tumor multiplicity, tumor volume, and histological grade were determined at 10 weeks after AdeCre administration. Cell proliferation was monitored by Ki67 immunohistochemistry at 4 and 10 weeks after AdeCre administration. At 10 weeks, female mice had more than twice the number of tumors evident on the surface of the lungs than male mice; ovariectomy eliminated this sex difference. The estrogen treatment significantly increased tumor number and volume in ovariectomized females and in males. Histological character of the tumors ranged from adenoma to adenocarcinoma. Ovary-intact females exhibited higher grade tumors than ovariectomized females or males. Progression to higher histological grade was stimulated by estrogen in male mice but not in ovariectomized females. At 10 weeks after AdeCre administration, tumor cell Ki67 labeling varied widely, precluding assessment of an estrogen effect; however, at 4 weeks, Ki67 labeling of lung parenchymal cells was increased 3.5-fold by estrogen. In conclusion, estrogen acts as a promoter for lung adenocarcinoma in a genetically defined lung cancer model; estrogen-induced cell proliferation in the oncogene-initiated cells is likely to play a role in this tumor promoter activity. PMID- 18509001 TI - Endogenous sex hormones and endometrial cancer risk in women in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - Epidemiological data show that reproductive and hormonal factors are involved in the etiology of endometrial cancer, but there is little data on the association with endogenous sex hormone levels. We analyzed the association between prediagnostic serum concentrations of sex steroids and endometrial cancer risk in the European Prospective Investigation into Cancer and Nutrition using a nested case-control design of 247 incident endometrial cancer cases and 481 controls, matched on center, menopausal status, age, variables relating to blood collection, and, for premenopausal women, phase of menstrual cycle. Using conditional regression analysis, endometrial cancer risk among postmenopausal women was positively associated with increasing levels of total testosterone, free testosterone, estrone, total estradiol, and free estradiol. The odds ratios (ORs) for the highest versus lowest tertile were 2.66 (95% confidence interval (CI) 1.50-4.72; P=0.002 for a continuous linear trend) for estrone, 2.07 (95% CI 1.20-3.60; P=0.001) for estradiol, and 1.66 (95% CI 0.98-2.82; P=0.001) for free estradiol. For total and free testosterone, ORs for the highest versus lowest tertile were 1.44 (95% CI 0.88-2.36; P=0.05) and 2.05 (95% CI 1.23-3.42; P=0.005) respectively. Androstenedione and dehydroepiandrosterone sulfate were not associated with risk. Sex hormone-binding globulin was significantly inversely associated with risk (OR for the highest versus lowest tertile was 0.57, 95% CI 0.34-0.95; P=0.004). In premenopausal women, serum sex hormone concentrations were not clearly associated with endometrial cancer risk, but numbers were too small to draw firm conclusions. In conclusion, relatively high blood concentrations of estrogens and free testosterone are associated with an increased endometrial cancer risk in postmenopausal women. PMID- 18509002 TI - Cytotoxic effects of a novel pyrazolopyrimidine derivative entrapped in liposomes in anaplastic thyroid cancer cells in vitro and in xenograft tumors in vivo. AB - In this study, we evaluated the activity of two novel pyrazolopyrimidine derivatives (Si 34 and Si 35) against ARO cells, a human anaplastic thyroid cancer cell line. ARO cells exposed to different concentrations of the drugs showed a reduced growth rate and an increase of mortality. After 72 h incubation, doses of 5 and 10 microM Si 34 determined a decrease of cell counts by approximately 25% and approximately 75% compared with those of control cells respectively. Similar findings were observed using Si 35. Treatment with both Si 34 and Si 35 at 10 microM increased cell mortality also ( approximately 29% and approximately 18% respectively). At these concentrations, a decrease in cyclin D1 levels was observed. To improve the biopharmaceutical properties, a liposome formulation was prepared. When entrapped in unilamellar liposomes, Si 34 exerted its cytotoxic effects even at lower doses (maximal inhibition at 5 microM) and after shorter incubation time (48 h) either in ARO or other thyroid cancer cell lines. The effects were associated with weak apoptotic death. Inhibition of epidermal growth factor-stimulated src and ERK phosphorylation, as well as reduction of migration properties of ARO cells was also observed. Moreover, the growth of tumor xenografts induced in severe combined immunodeficiency (SCID) mice was inhibited by i.v. administration of 25-50 mg/kg of the drug liposomal formulation. In conclusion, the liposomal preparation of this novel pyrazolopyrimidine derivative appears to be a promising tool for the treatment of anaplasic thyroid cancer. PMID- 18509003 TI - BRAFV600E mutation, but not RET/PTC rearrangements, is correlated with a lower expression of both thyroperoxidase and sodium iodide symporter genes in papillary thyroid cancer. AB - A low sodium iodide symporter (NIS) expression has been shown in papillary thyroid carcinomas (PTCs) harboring the BRAFV600E mutation. In the present study, we analyzed the mRNA expression of thyroid differentiation genes, glucose transporter (GLUT)-1 and GLUT-3, in 78 PTCs according to the presence of BRAFV600E or RET/PTC rearrangements. We found BRAFV600E and RET/PTC rearrangements in 35.8 and 19.4% of PTCs respectively. The mRNA expression of NIS and thyroperoxidase (TPO) genes were significantly lower (P<0.0001 and P=0.004 respectively) in BRAFV600E-positive PTC with respect to non-mutated samples. In support of this result, immunohistochemistry showed that the percentage of NIS positive cells was significantly lower (P=0.005) in BRAFV600E-mutated PTC (mean 53.5%) than in negative cases (mean 72.6%). In contrast, no difference either in NIS or in any other thyroid differentiation genes' mRNA expression was found in PTC with or without RET/PTC rearrangements. When GLUT-1 and GLUT-3 mRNA expression was considered, no correlation was found either in BRAFV600E- nor in RET/PTC-mutated cases. In conclusion, this study confirmed the presence of a genetic alteration of BRAF and/or RET oncogenes in 64% of PTC cases and revealed a significant correlation of BRAFV600E mutation with a lower expression of both NIS and TPO. This latter finding could indicate that an early dedifferentiation process is present at the molecular level in BRAFV600E-mutated PTC, thus suggesting that the previously demonstrated poor prognostic significance of BRAFV600E mutation could be related to the dedifferentiation process more than to a more advanced stage at diagnosis. PMID- 18509004 TI - Interactions between sphingosine-1-phosphate and vascular endothelial growth factor signalling in ML-1 follicular thyroid carcinoma cells. AB - Sphingosine-1-phosphate (S1P) induces migration of human ML-1 thyroid follicular cancer cells and inhibits migration of human FRO anaplastic thyroid cancer cells. As tumour cells often secrete vascular endothelial growth factor (VEGF), we investigated a possible interaction between S1P and VEGF signalling in the regulation of thyroid tumour cell migration. We found that both ML-1 and FRO cells secreted VEGF-A ( approximately 3.6 and <0.1 ng/10(6) cells/day respectively) and VEGF-C ( approximately 3.0 and 0.14 ng/10(6) cells/day respectively). S1P stimulated VEGF-A secretion in both cell lines, and blocking S1P receptors 1, 2 and 3 attenuated the S1P-evoked secretion of VEGF-A. Neither TSH nor insulin affected the amount of secreted VEGF-A or -C in ML-1 cells, while simultaneous stimulation with insulin and S1P increased VEGF-C secretion in FRO cells. Both cell lines expressed VEGF receptor 2 (VEGFR-2) mRNA and proteins. Serum-evoked migration of both ML-1 and FRO cells was attenuated when VEGFR-2 was inhibited. Moreover, inhibiting VEGFR-2 in ML-1 cells resulted in a rapid downregulation of S1P1 mRNA expression and S1P1 protein levels, suppression of S1P-induced migration and a decrease in S1P-induced Akt phosphorylation. A VEGF neutralizing antibody also reduced S1P-induced migration. In ML-1 cells, S1P phosphorylated VEGFR-2. In addition, VEGFR-2 inhibition resulted in the upregulation of S1P3 mRNA within 24 h, but a significant increase in S1P3 protein levels was not observed. VEGFR-2 inhibition, but not a VEGF-neutralizing antibody, reduced ML-1 cell proliferation independently of S1P stimulation. The results indicate a complex interaction between S1P and VEGFR-2 in ML-1 cells, particularly in regulating migratory responses. PMID- 18509005 TI - Peroxisome proliferator-activated receptor gamma inhibits follicular and anaplastic thyroid carcinoma cells growth by upregulating p21Cip1/WAF1 gene in a Sp1-dependent manner. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) has been demonstrated to be anti-neoplastic against various human tumors. The aim of this study was to delineate the molecular mechanism underlying PPARgamma ligand rosiglitazone (BRL) antiproliferative effects in follicular WRO and anaplastic FRO human thyroid carcinoma cells. BRL upregulated the p21Cip1/WAF1 levels in the two thyroid cancer cells, while did not modify the p53 protein content. Different evidences indicate that the p21Cip1/WAF1 upregulation by BRL requires a functional PPARgamma, since it was reversed by silencing PPARgamma and pretreatment with GW9662, an irreversible PPARgamma antagonist. Transient transfection assays showed that BRL triggered the transcriptional activity of p21Cip1/WAF1 promoter gene in a p53-independent way, being a p21Cip1/WAF1 promoter construct deleted in the p53 sites still activated by BRL. The Sp1 inhibitor mithramycin silenced the p21Cip1/WAF1 promoter activity suggesting an important role of Sp1 in mediating BRL activation. The electrophoretic mobility shift and chromatin immunoprecipitation (ChIP) assays evidenced a functional interaction between PPARgamma and Sp1 in regulating p21Cip1/WAF1. Intriguingly, ChIP analysis revealed in the p21Cip1/WAF1 gene promoter an increased recruitment of the RNA Pol II associated with an increased histone H3 acetylation and a reduced H3 methylation. The biological event, consistent with PPARgamma-induced WRO and FRO cell growth inhibition, was reversed by p21Cip1/WAF1 antisense oligonucleotides and was confirmed by increasing the PPARgamma expression, suggesting a crucial role exerted by p21Cip1/WAF1 in PPARgamma action. Our results further candidate BRL as a potential agent able to inhibit tumor progression of follicular and anaplastic thyroid carcinoma. PMID- 18509006 TI - Efficacy of a dopamine-somatostatin chimeric molecule, BIM-23A760, in the control of cell growth from primary cultures of human non-functioning pituitary adenomas: a multi-center study. AB - Dopamine D2 and somatostatin receptors (sstrs) were reported to affect non functioning pituitary adenoma (NFPA) proliferation in vitro. However, the reported results differ according to the experimental conditions used. We established an experimental protocol allowing reproducible evaluation of NFPA cell proliferation in vitro, to test and compare the antiproliferative effects of dopamine and somatostatin analogs (alone or in combination) with the activity of the dopamine-somatostatin chimeric molecule BIM-23A760. The protocol was utilized by four independent laboratories, studying 38 fibroblast-deprived NFPA cell cultures. Cells were characterized for GH, POMC, sstr1-sstr5, total dopamine D2 receptor (D2R) (in all cases), and D2 receptor long and short isoforms (in 15 out of 38 cases) mRNA expression and for alpha-subunit, LH, and FSH release. D2R, sstr3, and sstr2 mRNAs were consistently observed, with the dominant expression of D2R (2.9+/-2.6 copy/copy beta-glucuronidase; mean+/-s.e.m.), when compared with sstr3 and sstr2 (0.6+/-1.0 and 0.3+/-0.6 respectively). BIM-23A760, a molecule with high affinity for D2R and sstr2, significantly inhibited [3H]thymidine incorporation in 23 out of 38 (60%) NFPA cultures (EC50=1.2 pM and Emax=-33.6+/-3.7%). BIM-23A760 effects were similar to those induced by the selective D2R agonist cabergoline that showed a statistically significant inhibition in 18 out of 27 tumors (compared with a significant inhibition obtained in 17 out of 27 tumors using BIM-23A760, in the same subgroup of adenomas analyzed), while octreotide was effective in 13 out of 27 cases. In conclusion, superimposable data generated in four independent laboratories using a standardized protocol demonstrate that, in vitro, chimeric dopamine/sstr agonists are effective in inhibiting cell proliferation in two-thirds of NFPAs. PMID- 18509007 TI - Predicting values of lipids and white blood cell count for all-site cancer in type 2 diabetes. AB - Type 2 diabetic patients have increased cancer risk. We developed and validated an all-site cancer risk score in a prospective cohort of 7374 Chinese type 2 diabetic patients free of known history of cancer at enrolment, using split-half validation. Spline Cox model was used to detect common risk factors of cancer and to guide linear transformation of non-linear risk factors. After a median follow up period of 5.45 years, 365 patients (4.95%) developed cancer. Body mass index (BMI; <24.0 or > or =27.6 kg/m2), triglyceride (> or =0.81 to <1.41 mmol/l), high density lipoprotein cholesterol (<0.9 or > or =1.8 mmol/l), total cholesterol (<4.3 mmol/l) and white blood cell (WBC) count (<5.8x10(9) count per litre) were associated with increased cancer risks and exhibited non-linear relationships. We further linear transformed these terms for selection using backward Cox regression (P<0.05 for stay) in the training dataset. In the test dataset, calibration was checked using Hosmer-Lemeshow test and discrimination checked using area under receiver operating characteristic curve. In addition to age and current smoking, only linear-transformed total cholesterol and WBC count were selected. The risk score was 0.0488xage (years)-0.5810xtotal cholesterol (mmol/l, coded to 4.3 if >4.3)-0.3596xWBC count (10(9) counts/l, 5.8 if >5.8)+0.6390xcurrent smoking status (1 if yes). The 5-year probability of cancer was 1-0.9590(EXP(0.9382x(RISK SCORE+1.5903))). The predicted cancer probability was not significantly different from the observed cancer probability during the 5 year follow-up. The adjusted area under receiver operating characteristic curve was 0.712. In conclusion, BMI, lipids and WBC count have predicting values for cancer. PMID- 18509008 TI - Methylation of the p16INK4A promoter is associated with malignant behavior in abdominal extra-adrenal paragangliomas but not pheochromocytomas. AB - Pheochromocytomas and abdominal extra-adrenal paragangliomas are related to endocrine tumors of the sympathetic nervous system. Studies in animal models have shown that inactivation of the products of the cyclin dependent kinase inhibitor 2A (CDKN2A) gene locus, p16INK4A and p14ARF, promotes the development of pheochromocytoma, especially in malignant form. The present study evaluated the involvement of CDKN2A in human pheochromocytomas and abdominal extra-adrenal paragangliomas from 55 patients. Promoter methylation was assessed using quantitative Pyrosequencing and methylation-specific PCR, and mRNA expression was measured by quantitative real-time PCR. For p16, western blot analysis and sequencing were also performed. succinate dehydrogenase complex subunit B (SDHB) sequencing analysis included extra-adrenal paragangliomas, all tumors classified as malignant, and cases diagnosed at 30 years or younger. The p16INK4A promoter was heavily methylated in a subset of paragangliomas, and this was significantly associated with malignancy (P<0.0043) and SDHB mutation (P<0.002). p16INK4A mRNA expression showed moderate suppression in malignant cases (P<0.05). In contrast, very little p14ARF promoter methylation was seen and there was no significant difference in p14ARF expression between tumors and normal samples. The p16 protein expression was reduced in 16 tumors, and sequence variations were observed in four tumors including the missense mutation A57V and the single nucleotide polymorphism (SNP) A148T. The results suggest that p16INK4A, and not p14ARF, is a subject of frequent involvement in these tumors. Importantly, hypermethylation of the p16INK4A promoter was significantly associated with malignancy and metastasis, and SDHB gene mutations. This finding suggests an etiological link and could provide a clinical utility for diagnostic purposes. PMID- 18509009 TI - Mitotane increases the radiotherapy inhibitory effect and induces G2-arrest in combined treatment on both H295R and SW13 adrenocortical cell lines. AB - Mitotane, 1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)ethane (o,p'-DDD) is an agent with adrenotoxic effect, which is able to block cortisol synthesis. This drug and radiotherapy are used also in adrenal cancer treatment even if their biological action in this neoplasia remains unknown. We investigated the effects of o,p'-DDD and ionizing radiations (IR) on cell growth inhibition and cell cycle perturbation in H295R and SW13 adrenocortical cancer cells. Both cell lines were irradiated at a 6 Gy dose and were treated with o,p'-DDD 10(-5) M separately and with IR/o,p'-DDD in combination. This combination treatment induced an irreversible inhibition of cell growth in both adrenocortical cancer cells. Cell cycle analysis showed that IR alone and IR/o,p'-DDD in combination induced the cell accumulation in the G2 phase. At 120 h after IR, the cells were able to recover the IR-induced G2 block while cells treated with IR/o,p'-DDD were still arrested in G2 phase. In order to study the molecular mechanism involved in the G2 irreversible arrest, we have considered the H295R cell line showing the highest inhibition of cell proliferation associated with a noteworthy G2 arrest. In these cells, cyclin B1 and Cdk2 proteins were examined by western blot and Cdk2 kinase activity measured by assay kit. The H295R cells treated with IR/o,p' DDD shared an increase in cyclin B1 amount as the coimmunoprecipitation of Cdc2 cyclin B1 complex. The kinase activity also shows an increase in the treated cells with combination therapy. Moreover, in these cells, sequence analysis of p53 revealed a large deletion of exons 8 and 9. The same irreversible block on G2 phase, induced by IR/o,p'-DDD treatment, happened in H295R cells with restored wild-type p53 suggesting that this mechanism is not mediated by p53 pathway. PMID- 18509010 TI - Targeted therapy for adrenocortical tumors in transgenic mice through their LH receptor by Hecate-human chorionic gonadotropin beta conjugate. AB - Novel strategies are needed for the treatment of adrenocortical tumors that are usually resistant to chemotherapy. Hecate, a 23-amino acid lytic peptide, was conjugated to the 15-amino acid (81-95) fragment of the human chorionic gonadotropin beta (CGbeta) chain, which would selectively kill cancer cells expressing the LH receptor (LHR) sparing the normal ones with LHR. To prove the principle that Hecate-CGbeta conjugate may eradicate tumors ectopically expressing plasma membrane receptors, transgenic (TG) inhibin alpha-subunit promoter (inhalpha)/Simian Virus 40 T-antigen mice, expressing LHR in their adrenal gland tumors, were used as the experimental model. Wild-type control littermates and TG mice with adrenal tumors were treated with either Hecate or Hecate-CGbeta conjugate at the age of 6.5 months for 3 weeks and killed 7 days after the last treatment. The Hecate-CGbeta conjugate reduced the adrenal tumor burden significantly in TG male but not in female mice, in comparison with Hecate treated mice. Hecate-CGbeta conjugate treatment did not affect normal adrenocortical function as the serum corticosterone level between Hecate and Hecate-CGbeta conjugate groups were similar. The mRNA and protein expressions of GATA-4 and LHR colocalized only in tumor area, and a significant downregulation of gene expression was found after the Hecate-CGbeta conjugate in comparison with Hecate- and/or non-treated adrenal tumors by western blotting. This finding provides evidence for a selective destruction of the tumor cells by the Hecate CGbeta conjugate. Hereby, our findings support the principle that Hecate-CGbeta conjugate is able to specifically destroy tumor cells that ectopically express LHR. PMID- 18509011 TI - Protein microarrays address the elephant in the room. PMID- 18509012 TI - MicroRNA signatures as diagnostic and therapeutic targets. PMID- 18509013 TI - Effect of collection tube type and preanalytical handling on myeloperoxidase concentrations. AB - BACKGROUND: Myeloperoxidase (MPO) has shown potential as a marker for cardiovascular disease. Limited studies have been published with a variety of sample types, resulting in a wide range of MPO values. Little is known or understood about the impact of collection tube type and preanalytical handling of specimens for MPO determination. METHOD: MPO concentration was determined by use of the ARCHITECT(R) MPO research use assay, which is currently under development. Samples were collected into multiple anticoagulant collection tubes from donors and patients presenting to the emergency department with symptoms of acute coronary syndromes. Whole blood was stored on ice or at room temperature for predetermined time periods. We also evaluated serum and plasma after centrifugation followed by storage at room temperature, 2-8 degrees C, and below 10 degrees C. RESULTS: Baseline sample concentrations were dependent on collection tube type as well as handling conditions. MPO concentrations were consistently higher in samples collected in serum and heparin plasma tubes than in samples in EDTA or citrate tubes. Spike recovery was acceptable in all sera and plasma tested, indicating that the increased MPO concentrations were not due directly to an anticoagulant interference. CONCLUSIONS: The collection tube type and preanalytical handling are critical for accurate and consistent MPO measurement. The preferred anticoagulant and tubes are the EDTA or EDTA plasma preparation tube. MPO concentrations in samples collected in these tubes are stable before centrifugation as whole blood as well as plasma after processing. PMID- 18509014 TI - Rapid determination of monozygous twinning with a microfabricated capillary array electrophoresis genetic-analysis device. AB - BACKGROUND: Microfabricated genetic-analysis devices have great potential for delivering complex clinical diagnostic technology to the point of care. As a demonstration of the potential of these devices, we used a microfabricated capillary array electrophoresis (microCAE) instrument to rapidly characterize the familial and genotypic relationship of twins who had been assigned fraternal (dizygous) status at birth. METHODS: We extracted the genomic DNA from buccal samples collected from the twin sons, the parents, another sibling, and an unrelated control individual. We then carried out multiplex PCR amplification of sequences at 16 short tandem repeat loci commonly used in forensic identity testing. We simultaneously separated the amplicons from all of the individuals on a microCAE device and fluorescently detected the amplicons with single-base resolution in <30 min. RESULTS: The genotypic analysis confirmed the identical status of the twins and revealed, in conjunction with the medical data, that their twin status arose from the rarer dichorionic, diamniotic process. CONCLUSIONS: The ability to rapidly analyze complex genetic samples with microCAE devices demonstrates that this approach can help meet the growing need for rapid genetics-based diagnostics. PMID- 18509015 TI - Importance of chemical reduction in plasma and serum homocysteine analysis. PMID- 18509016 TI - Improved HPLC analysis of serum 7alpha-hydroxycholest-4-en-3-one, a marker of bile acid malabsorption. PMID- 18509017 TI - Unexpected hemoglobin A results after an erythrocyte exchange: importance of specimen mixing. PMID- 18509018 TI - Insufficient standardization of a direct carbohydrate-deficient transferrin immunoassay. PMID- 18509019 TI - Influence of L-thyroxine therapy on parathyroid hormone concentrations. PMID- 18509020 TI - Acute variation of osteocalcin and parathyroid hormone in athletes after running a half-marathon. PMID- 18509021 TI - Rapid real-time PCR detection of HPdel directly from diluted blood samples. PMID- 18509022 TI - The N-type voltage-gated calcium channel: when a neuron reads a map. PMID- 18509023 TI - Dissociating the role of the orbitofrontal cortex and the striatum in the computation of goal values and prediction errors. AB - To make sound economic decisions, the brain needs to compute several different value-related signals. These include goal values that measure the predicted reward that results from the outcome generated by each of the actions under consideration, decision values that measure the net value of taking the different actions, and prediction errors that measure deviations from individuals' previous reward expectations. We used functional magnetic resonance imaging and a novel decision-making paradigm to dissociate the neural basis of these three computations. Our results show that they are supported by different neural substrates: goal values are correlated with activity in the medial orbitofrontal cortex, decision values are correlated with activity in the central orbitofrontal cortex, and prediction errors are correlated with activity in the ventral striatum. PMID- 18509024 TI - Mechanisms underlying functional changes in the primary motor cortex ipsilateral to an active hand. AB - Performance of a unimanual hand motor task results in functional changes in both primary motor cortices (M1(ipsilateral) and M1(contralateral)). The neuronal mechanisms controlling the corticospinal output originated in M1(ipsilateral) and the resting hand during a unimanual task remain unclear. Here, we assessed functional changes within M1(ipsilateral) and in interhemispheric inhibition (IHI) associated with parametric increases in unimanual force. We measured motor evoked potential (MEP) recruitment curves (RCs) and short-interval intracortical inhibition (SICI) in M1(ipsilateral), IHI from M1(contralateral) to M1(ipsilateral), and the influence of IHI over SICI using transcranial magnetic stimulation at rest and during 10, 30, and 70% of maximal right wrist flexion force. EMG from the left resting flexor carpi radialis (FCR) muscle was comparable across conditions. Left FCR MEP RCs increased, and SICI decreased with increasing right wrist force. Activity-dependent (rest and 10, 30, and 70%) left FCR maximal MEP size correlated with absolute changes in SICI. IHI decreased with increasing force at matched conditioned MEP amplitudes. IHI and SICI were inversely correlated at increasing forces. In the presence of IHI, SICI decreased at rest and 70% force. In summary, we found activity-dependent changes in (1) SICI in M1(ipsilateral), (2) IHI from M1(contralateral) to M1(ipsilateral), and (3) the influence of IHI over SICI in the left resting hand during force generation by the right hand. Our findings indicate that interactions between GABAergic intracortical circuits mediating SICI and interhemispheric glutamatergic projections between M1s contribute to control activity-dependent changes in corticospinal output to a resting hand during force generation by the opposite hand. PMID- 18509025 TI - An essential role for Frizzled5 in neuronal survival in the parafascicular nucleus of the thalamus. AB - Frizzled5 (Fz5), a putative Wnt receptor, is expressed in the retina, hypothalamus, and the parafascicular nucleus (PFN) of the thalamus. By constructing Fz5 alleles in which beta-galactosidase replaces Fz5 or in which Cre mediated recombination replaces Fz5 with alkaline phosphatase, we observe that Fz5 is required continuously and in a cell autonomous manner for the survival of adult PFN neurons, but is not required for proliferation, migration, or axonal growth and targeting of developing PFN neurons. A motor phenotype associated with loss of Fz5 establishes a role for the PFN in sensorimotor coordination. Transcripts coding for Wnt9b, the likely Fz5 ligand in vivo, and beta-catenin, a mediator of canonical Wnt signaling, are both downregulated in the Fz5(-/-) PFN, implying a positive feedback mechanism in which Wnt signaling is required to maintain the expression of Wnt signaling components. These data suggest that defects in Wnt-Frizzled signaling could be the cause of neuronal loss in degenerative CNS diseases. PMID- 18509026 TI - Organization of the Arp2/3 complex in hippocampal spines. AB - Changes in the morphology of a dendritic spine require remodeling of its actin based cytoskeleton. Biochemical mechanisms underlying actin remodeling have been studied extensively, but little is known about the physical organization of the actin-binding proteins that mediate remodeling in spines. Long-term potentiation inducing stimuli trigger expansion of the spine head, suggesting local extension and branching of actin filaments. Because filament branching requires the Arp2/3 complex, we used quantitative immunoelectron microscopy to elucidate the organization of ARPC-2 (Arp2/3 complex subunit 2), an essential component of the complex. Our data from CA1 hippocampus indicate that Arp2/3 concentrates within spines in a previously unrecognized torroidal domain, apparently specialized to mediate actin filament branching. PMID- 18509027 TI - The best disease-linked Cl- channel hBest1 regulates Ca V 1 (L-type) Ca2+ channels via src-homology-binding domains. AB - Mutations in the bestrophin-1 (Best1) gene are linked to several kinds of macular degeneration in both humans and dogs. Although bestrophins have been shown clearly to be Cl(-) ion channels, it is controversial whether Cl(-) channel dysfunction can explain the diseases. It has been suggested that bestrophins are multifunctional proteins: they may regulate voltage-gated Ca(2+) channels in addition to functioning as Cl(-) channels. Here, we show that human Best1 gene (hBest1) differentially modulates Ca(V)1.3 (L-type) voltage-gated Ca(2+) channels through association with the Ca(V)beta subunit. In transfected human embryonic kidney 293 cells, hBest1 inhibited Ca(V)1.3. Inhibition of Ca(V)1.3 was not observed in the absence of the beta subunit. Also, the hBest1 C terminus binds to Ca(V)beta subunits, suggesting that the effect of hBest1 was mediated by the Ca(V)beta subunit. The region of hBest1 responsible for the effect was localized to a region (amino acids 330-370) in the cytoplasmic C terminus that contains a predicted src-homology-binding domain that is not present in other bestrophin subtypes. Mutation of Pro(330) and Pro(334) abolished the effects of hBest1 on Ca(V)1.3. The effect was specific to hBest1; it was not observed with mouse Best1 (mBest1), mBest2, or mBest3. Wild-type hBest1 and the disease-causing mutants R92S, G299R, and D312N inhibited Ca(V) currents the same amount, whereas the A146K and G222E mutants were less effective. We propose that hBest1 regulates Ca(V) channels by interacting with the Ca(V)beta subunit and altering channel availability. Our findings reveal a novel function of bestrophin in regulation of Ca(V) channels and suggest a possible mechanism for the role of hBest1 in macular degeneration. PMID- 18509028 TI - Opposing patterns of signaling activation in dopamine D1 and D2 receptor expressing striatal neurons in response to cocaine and haloperidol. AB - Psychostimulants and other drugs of abuse activate extracellular signal-regulated kinase (ERK) in the striatum, through combined stimulation of dopamine D(1) receptors (D1Rs) and glutamate NMDA receptors. Antipsychotic drugs activate similar signaling proteins in the striatum by blocking dopamine D(2) receptors (D2Rs). However, the neurons in which these pathways are activated by psychotropic drugs are not precisely identified. We used transgenic mice, in which enhanced green fluorescent protein (EGFP) expression was driven by D1R promoter (drd1a-EGFP) or D2R promoter (drd2-EGFP). We confirmed the expression of drd1a-EGFP in striatonigral and drd2-EGFP in striatopallidal neurons. Drd2-EGFP was also expressed in cholinergic interneurons, whereas no expression of either promoter was detected in GABAergic interneurons. Acute cocaine treatment increased phosphorylation of ERK and its direct or indirect nuclear targets, mitogen- and stress-activated kinase-1 (MSK1) and histone H3, exclusively in D1R expressing output neurons in the dorsal striatum and nucleus accumbens. Cocaine induced expression of c-Fos and Zif268 predominated in D1R-expressing neurons but was also observed in D2R-expressing neurons. One week after repeated cocaine administration, cocaine-induced signaling responses were decreased, with the exception of enhanced ERK phosphorylation in dorsal striatum. The responses remained confined to D1R neurons. In contrast, acute haloperidol injection activated phosphorylation of ERK, MSK1, and H3 only in D2R neurons and induced c fos and zif268 predominantly in these neurons. Our results demonstrate that cocaine and haloperidol specifically activate signaling pathways in two completely segregated populations of striatal output neurons, providing direct evidence for the selective mechanisms by which these drugs exert their long-term effects. PMID- 18509029 TI - Transient spine expansion and learning-induced plasticity in layer 1 primary motor cortex. AB - Experience-dependent regulation of synaptic strength in the horizontal connections in layer 1 of the primary motor cortex is likely to play an important role in motor learning. Dendritic spines, the primary sites of excitatory synapses in the brain, are known to change shape in response to various experimental stimuli. We used a rat motor learning model to examine connection strength via field recordings in slices and confocal imaging of labeled spines to explore changes induced solely by learning a simple motor task. We report that motor learning increases response size, while transiently occluding long-term potentiation (LTP) and increasing spine width in layer 1. This demonstrates learning-induced changes in behavior, synaptic responses, and structure in the same animal, suggesting that an LTP-like process in the motor cortex mediates the initial learning of a skilled task. PMID- 18509030 TI - Feedback from horizontal cells to rod photoreceptors in vertebrate retina. AB - Retinal horizontal cells (HCs) provide negative feedback to cones, but, largely because annular illumination fails to evoke a depolarizing response in rods, it is widely believed that there is no feedback from HCs to rods. However, feedback from HCs to cones involves small changes in the calcium current (I(Ca)) that do not always generate detectable depolarizing responses. We therefore recorded I(Ca) directly from rods to test whether they were modulated by feedback from HCs. To circumvent problems presented by overlapping receptive fields of HCs and rods, we manipulated the membrane potential of voltage-clamped HCs while simultaneously recording from rods in a salamander retinal slice preparation. Like HC feedback in cones, hyperpolarizing HCs from -14 to -54, -84, and -104 mV increased the amplitude of I(Ca) recorded from synaptically connected rods and caused hyperpolarizing shifts in I(Ca) voltage dependence. These effects were blocked by supplementing the bicarbonate-buffered saline solution with HEPES. In rods lacking light-responsive outer segments, hyperpolarizing neighboring HCs with light caused a negative activation shift and increased the amplitude of I(Ca). These changes in I(Ca) were blocked by HEPES and by inhibiting HC light responses with a glutamate antagonist, indicating that they were caused by HC feedback. These results show that rods, like cones, receive negative feedback from HCs that regulates the amplitude and voltage dependence of I(Ca). HC-to-rod feedback counters light-evoked decreases in synaptic output and thus shapes the transmission of rod responses to downstream visual neurons. PMID- 18509031 TI - Low-frequency local field potentials and spikes in primary visual cortex convey independent visual information. AB - Local field potentials (LFPs) reflect subthreshold integrative processes that complement spike train measures. However, little is yet known about the differences between how LFPs and spikes encode rich naturalistic sensory stimuli. We addressed this question by recording LFPs and spikes from the primary visual cortex of anesthetized macaques while presenting a color movie. We then determined how the power of LFPs and spikes at different frequencies represents the visual features in the movie. We found that the most informative LFP frequency ranges were 1-8 and 60-100 Hz. LFPs in the range of 12-40 Hz carried little information about the stimulus, and may primarily reflect neuromodulatory inputs. Spike power was informative only at frequencies <12 Hz. We further quantified "signal correlations" (correlations in the trial-averaged power response to different stimuli) and "noise correlations" (trial-by-trial correlations in the fluctuations around the average) of LFPs and spikes recorded from the same electrode. We found positive signal correlation between high-gamma LFPs (60-100 Hz) and spikes, as well as strong positive signal correlation within high-gamma LFPs, suggesting that high-gamma LFPs and spikes are generated within the same network. LFPs <24 Hz shared strong positive noise correlations, indicating that they are influenced by a common source, such as a diffuse neuromodulatory input. LFPs <40 Hz showed very little signal and noise correlations with LFPs >40 Hz and with spikes, suggesting that low-frequency LFPs reflect neural processes that in natural conditions are fully decoupled from those giving rise to spikes and to high-gamma LFPs. PMID- 18509032 TI - Accurate balance of the polarity kinase MARK2/Par-1 is required for proper cortical neuronal migration. AB - Radial neuronal migration is key in structuring the layered cortex. Here we studied the role of MARK2/Par-1 in this process. The dual name stands for the MAP/microtubule affinity-regulating kinase 2 (MARK2) and the known polarity kinase 1 (Par-1). Reduced MARK2 levels using in utero electroporation resulted in multipolar neurons stalled at the intermediate zone border. Reintroduction of the wild-type kinase postmitotically improved neuronal migration. Our results indicated that reduction in MARK2 affected centrosomal dynamics in migrating neurons of the cerebral cortex. Increased MARK2 has been shown to destabilize microtubules, and here we show for the first time that reduced MARK2 stabilized microtubules in primary cultured neurons. Kinase-independent activity permitted multipolar-to-bipolar transition but did not restore proper migration. Increased MARK2 levels resulted in a different phenotype, which is loss of neuronal polarity. MARK2 kinase activity reduction hindered migration in the developing brain, which was rescued by increasing kinase activity. Our results stress the necessity of maintaining dynamic microtubules for proper neuronal migration. Furthermore, the exact requirements for MARK2 and its kinase activity vary during the course of neuronal migration. Collectively, our results stress the requirements for the different roles of MARK2 during neuronal migration. PMID- 18509033 TI - Stress induces a switch of intracellular signaling in sensory neurons in a model of generalized pain. AB - Stress dramatically exacerbates pain in diseases such as fibromyalgia and rheumatoid arthritis, but the underlying mechanisms are unknown. We tested the hypothesis that stress causes generalized hyperalgesia by enhancing pronociceptive effects of immune mediators. Rats exposed to nonhabituating sound stress exhibited no change in mechanical nociceptive threshold, but showed a marked increase in hyperalgesia evoked by local injections of prostaglandin E(2) or epinephrine. This enhancement, which developed more than a week after exposure to stress, required concerted action of glucocorticoids and catecholamines at receptors located in the periphery on sensory afferents. The altered response to pronociceptive mediators involved a switch in coupling of their receptors from predominantly stimulatory to inhibitory G-proteins (G(s) to G(i)), and for prostaglandin E(2), emergence of novel dependence on protein kinase C epsilon. Thus, an important mechanism in generalized pain syndromes may be stress-induced coactivation of the hypothalamo-pituitary-adrenal and sympathoadrenal axes, causing a long-lasting alteration in intracellular signaling pathways, enabling normally innocuous levels of immune mediators to produce chronic hyperalgesia. PMID- 18509034 TI - Postsynaptic density-93 clusters Kv1 channels at axon initial segments independently of Caspr2. AB - Postsynaptic density-93 (PSD-93)/Chapsyn-110 is a PDZ (PSD-95/Discs large/zona occludens-1) domain-containing membrane-associated guanylate kinase (MAGUK) that functions as a scaffold to assemble channels, receptors, and other signaling proteins at cell membranes. PSD-93 is highly enriched at synapses, but mice lacking this protein have no synaptic structural abnormalities, probably because of overlapping expression and redundancy with other MAGUKs. Consequently, the function of PSD-93 is not well understood. Here, we show that PSD-93, but not other MAGUKs, is enriched at the axon initial segment (AIS), where it colocalizes with Kv1.1, Kv1.2, Kv1.4, and Kvbeta2 subunit-containing K(+) channels, Caspr2, and TAG-1 (transient axonal glycoprotein-1). When coexpressed with Kv1 channels in heterologous cells, PSD-93 induces formation of large cell-surface clusters. Knockdown of PSD-93 in cultured hippocampal neurons by RNA interference disrupted Kv1 channel localization at the AIS. Similarly, PSD-93-/- mice failed to cluster Kv1 channels at the AIS of cortical and hippocampal neurons. In contrast, Caspr2, which mediates Kv1 channel clustering at the juxtaparanode, is not required for localization of Kv1 channels at the AIS. These results show PSD-93 mediates AIS accumulation of Kv1 channels independently of Caspr2. PMID- 18509035 TI - Spine expansion and stabilization associated with long-term potentiation. AB - Stable expression of long-term synaptic plasticity is critical for the developmental refinement of neural circuits and for some forms of learning and memory. Although structural remodeling of dendritic spines is associated with the stable expression of long-term potentiation (LTP), the relationship between structural and physiological plasticity remains unclear. To define whether these two processes are related or distinct, we simultaneously monitored EPSPs and dendritic spines, using combined patch-clamp recording and two-photon time-lapse imaging in the same CA1 pyramidal neurons in acute hippocampal slices. We found that theta burst stimulation paired with postsynaptic spiking, which reliably induced LTP, also induced a rapid and persistent expansion of dendritic spines. Like LTP, this expansion was NMDA receptor dependent. Spine expansion occurred even when LTP was inhibited by postsynaptic inhibition of exocytosis or PKA (protein kinase A); however, under these conditions, the spine expansion was unstable and collapsed spontaneously. Furthermore, similar changes in LTP and spine expansion were observed when hippocampal neurons were treated with protein synthesis inhibitors. Like LTP, spine expansion was reversed by low-frequency stimulation (LFS) via a phosphatase-dependent mechanism, but only if the LFS was applied in a critical time window after induction. These results indicate that the initial expression of LTP and spine expansion is dissociable, but there is a high degree of mechanistic overlap between the stabilization of structural plasticity and LTP. PMID- 18509036 TI - The glutamate receptor-interacting protein family of GluR2-binding proteins is required for long-term synaptic depression expression in cerebellar Purkinje cells. AB - Glutamate receptor-interacting protein 1 (GRIP1) and GRIP2 are closely related proteins that bind GluR2-containing AMPA receptors and couple them to structural and signaling complexes in neurons. Cerebellar long-term synaptic depression (LTD) is a model system of synaptic plasticity that is expressed by persistent internalization of GluR2-containing AMPA receptors. Here, we show that genetic deletion of both GRIP1 and GRIP2 blocks LTD expression in primary cultures of mouse cerebellar neurons but that single deletion of either isoform allows LTD to occur. In GRIP1/2 double knock-out Purkinje cells, LTD can be fully rescued by a plasmid-driving expression of GRIP1 and partially rescued by a GRIP2 plasmid. These results indicate that the GRIP family comprises an essential molecular component for cerebellar LTD. PMID- 18509037 TI - Methamphetamine causes microglial activation in the brains of human abusers. AB - Methamphetamine is a popular addictive drug whose use is associated with multiple neuropsychiatric adverse events and toxic to the dopaminergic and serotonergic systems of the brain. Methamphetamine-induced neuropathology is associated with increased expression of microglial cells that are thought to participate in either pro-toxic or protective mechanisms in the brain. Although reactive microgliosis has been observed in animal models of methamphetamine neurotoxicity, no study has reported on the status of microglial activation in human methamphetamine abusers. The present study reports on 12 abstinent methamphetamine abusers and 12 age-, gender-, and education-matched control subjects who underwent positron emission tomography using a radiotracer for activated microglia, [(11)C](R)-(1-[2-chlorophenyl]-N-methyl-N-[1-methylpropyl]-3 isoquinoline carboxamide) ([(11)C](R)-PK11195). Compartment analysis was used to estimate quantitative levels of binding potentials of [(11)C](R)-PK11195 in brain regions with dopaminergic and/or serotonergic innervation. The mean levels of [(11)C](R)-PK11195 binding were higher in methamphetamine abusers than those in control subjects in all brain regions (>250% higher; p < 0.01 for all). In addition, the binding levels in the midbrain, striatum, thalamus, and orbitofrontal and insular cortices (p < 0.05) correlated inversely with the duration of methamphetamine abstinence. These results suggest that chronic self administration of methamphetamine can cause reactive microgliosis in the brains of human methamphetamine abusers, a level of activation that appears to subside over longer periods of abstinence. PMID- 18509038 TI - Cellular and behavioral interactions of gabapentin with alcohol dependence. AB - Gabapentin is a structural analog of GABA that has anticonvulsant properties. Despite the therapeutic efficacy of gabapentin, its molecular and cellular mechanisms of action are unclear. The GABAergic system in the central nucleus of the amygdala (CeA) plays an important role in regulating voluntary ethanol intake. Here, we investigated the effect of gabapentin on GABAergic transmission in CeA slices, on ethanol intake, and on an anxiety measure using animal models of ethanol dependence. Gabapentin increased the amplitudes of evoked GABA receptor-mediated IPSCs (GABA-IPSCs) in CeA neurons from nondependent rats, but decreased their amplitudes in CeA of ethanol-dependent rats. Gabapentin effects were blocked in the presence of a specific GABA(B) receptor antagonist. The sensitivity of the GABA-IPSCs to a GABA(B) receptor antagonist and an agonist was decreased after chronic ethanol, suggesting that ethanol-induced neuroadaptations of GABA(B) receptors associated with ethanol dependence may account for the differential effects of gabapentin after chronic ethanol. Systemic gabapentin reduced ethanol intake in dependent, but not in nondependent, rats and reversed the anxiogenic-like effects of ethanol abstinence using an acute dependence model. Gabapentin infused directly into the CeA also blocked dependence-induced elevation in operant ethanol responding. Collectively, these findings show that gabapentin reverses behavioral measures of ethanol dependence and, in turn, dependence reverses the effects of gabapentin on CeA neurons, and suggest that gabapentin represents a potential medication for treatment of alcoholism. PMID- 18509039 TI - Pronounced reduction of digit motor responses evoked from macaque ventral premotor cortex after reversible inactivation of the primary motor cortex hand area. AB - In common with other secondary motor areas, the macaque ventral premotor cortex (PMv) gives rise to corticospinal projections; it also makes numerous reciprocal corticocortical connections with the primary motor cortex (M1). Repetitive intracortical microstimulation (rICMS) of the PMv gives rise to movements of the hand and digits. To investigate whether these motor effects are dependent on the corticocortical interactions with M1, the effect of reversible inactivation of the M1 hand area was tested in three macaque monkeys with chronically implanted intracortical electrodes in the hand representations of M1 and PMv (rostral division, area F5). Monkeys were lightly sedated. Test EMG responses to rICMS were recorded from intrinsic hand muscles before and after microinjection of the GABA agonist muscimol in the M1 hand area. This not only greatly reduced EMG responses evoked from M1, but also reduced or abolished responses from F5, over a similar time course (20-50 min). Muscimol in M1 reduced the level of background EMG activity in the contralateral hand, which was paretic for several hours after injection. However, because EMG responses to direct activation of the corticospinal tract were significantly less affected than the responses to F5 stimulation, it is unlikely that reduced motoneuronal excitability explained the loss of the evoked responses from F5. Finally, muscimol injections in M1 greatly reduced the corticospinal volleys evoked by rICMS in F5. The results suggest that the motor effects evoked from F5 depend, at least in part, on corticocortical interactions with M1, leading to activation of M1 corticospinal outputs to hand muscles. PMID- 18509040 TI - Toll-like receptor 2 acts as a natural innate immune receptor to clear amyloid beta 1-42 and delay the cognitive decline in a mouse model of Alzheimer's disease. AB - Microglia are the immune cells of the brain, they are activated in the brain of Alzheimer's disease (AD) patients and mouse models of AD, and they express the innate immune receptor toll-like receptor 2 (TLR2). The present study investigated role of this receptor in the progression of AD-like pathologies. Here we show that amyloid beta (A beta) stimulates TLR2 expression in a small proportion of microglia. We then generated triple transgenic mice that are deficient in TLR2 from mice that harbor a mutant human presenelin 1 and a chimeric mouse/human amyloid precursor protein (APP) genes. TLR2 deficiency accelerated spatial and contextual memory impairments, which correlated with increased levels of A beta(1-42) and transforming growth factor beta1 in the brain. NMDA receptors 1 and 2A expression levels were also lower in the hippocampus of APP-TLR2(-/-) mice. Gene therapy in cells of the bone marrow using lentivirus constructs expressing TLR2 rescued the cognitive impairment of APP TLR2(-/-) mice. Indeed, lenti-green fluorescent protein/TLR2 treatment had beneficial effects by restoring the memory consolidation process disrupted by TLR2 deficiency in APP mice. These data suggest that TLR2 acts as an endogenous receptor for the clearance of toxic A beta by bone-marrow-derived immune cells. The cognitive decline is markedly accelerated in a context of TLR2 deficiency. Upregulating this innate immune receptor may then be considered as a potential new powerful therapeutic approach for AD. PMID- 18509041 TI - Cell-autonomous roles of ARX in cell proliferation and neuronal migration during corticogenesis. AB - The aristaless-related homeobox (ARX) gene has been implicated in a wide spectrum of disorders ranging from phenotypes with severe neuronal migration defects, such as lissencephaly, to mild forms of X-linked mental retardation without apparent brain abnormalities. To better understand its role in corticogenesis, we used in utero electroporation to knock down or overexpress ARX. We show here that targeted inhibition of ARX causes cortical progenitor cells to exit the cell cycle prematurely and impairs their migration toward the cortical plate. In contrast, ARX overexpression increases the length of the cell cycle. In addition, we report that RNA interference-mediated inactivation of ARX prevents cells from acquiring multipolar morphology in the subventricular and intermediate zones, resulting in decreased neuronal motility. In contrast, ARX overexpression appears to promote the development of tangentially oriented processes of cells in the subventricular and intermediate zones and affects radial migration of pyramidal neurons. We also demonstrate that the level of ARX expression is important for tangential migration of GABA-containing interneurons, because both inactivation and overexpression of the gene impair their migration from the ganglionic eminence. However, our data suggest that ARX is not directly involved in GABAergic cell fate specification. Overall, these results identify multiple and distinct cell-autonomous roles for ARX in corticogenesis. PMID- 18509042 TI - Noradrenergic innervation of the dorsal medial prefrontal cortex modulates hypothalamo-pituitary-adrenal responses to acute emotional stress. AB - The medial prefrontal cortex (mPFC) has been proposed to play a role in the inhibition of hypothalamo-pituitary-adrenal (HPA) responses to emotional stress via influences on neuroendocrine effector mechanisms housed in the paraventricular hypothalamic nucleus (PVH). Previous work also suggests an involvement of the locus ceruleus (LC) in behavioral and neuroendocrine responses to a variety of acute stressors. The LC issues a widespread set of noradrenergic projections, and its innervation of the prefrontal cortex plays an important role in the modulation of working memory and attention. Because these operations are likely to be critical for stimulus selection, evaluation, and comparison with past experience in mounting adaptive responses to emotional stress, it follows that the LC-to-mPFC pathway might also be involved in regulating HPA activity under such conditions. Therefore, in the present study, we assessed the effects of selectively ablating noradrenergic inputs into the mPFC, using the axonally transported catecholamine immunotoxin, saporin-conjugated antiserum to dopamine beta-hydroxylase, on acute restraint stress-induced activation of HPA output. Immunotoxin injections in the dorsal mPFC (centered in the prelimbic cortex) attenuated increments in restraint-induced Fos and corticotropin-releasing factor mRNA expression in the neurosecretory region of PVH, as well as HPA secretory responses. Stress-induced Fos expression in dorsal mPFC was enhanced after noradrenergic deafferentation and was negatively correlated with stress-induced PVH activation, independent of lesion status. These findings identify the LC as an upstream component of a circuitry providing for dorsal mPFC modulation of emotional stress-induced HPA activation. PMID- 18509043 TI - GPR56 regulates pial basement membrane integrity and cortical lamination. AB - GPR56 is a member of the family of adhesion G-protein-coupled receptors that have a large extracellular region containing a GPS (G-protein proteolytic site) domain. Loss-of-function mutations in the GPR56 gene cause a specific human brain malformation called bilateral frontoparietal polymicrogyria (BFPP). BFPP is a radiological diagnosis and its histopathology remains unclear. This study demonstrates that loss of the mouse Gpr56 gene leads to neuronal ectopia in the cerebral cortex, a cobblestone-like cortical malformation. There are four crucial events in the development of cobblestone cortex, namely defective pial basement membrane (BM), abnormal anchorage of radial glial endfeet, mislocalized Cajal Retzius cells, and neuronal overmigration. By detailed time course analysis, we reveal that the leading causal events are likely the breaches in the pial BM. We show further that GPR56 is present in abundance in radial glial endfeet. Furthermore, a putative ligand of GPR56 is localized in the marginal zone or overlying extracellular matrix. These observations provide compelling evidence that GPR56 functions in regulating pial BM integrity during cortical development. PMID- 18509044 TI - Zinc triggers microglial activation. AB - Microglia are resident immune cells of the CNS. When stimulated by infection, tissue injury, or other signals, microglia assume an activated, "ameboid" morphology and release matrix metalloproteinases, reactive oxygen species, and other proinflammatory factors. This innate immune response augments host defenses, but it can also contribute to neuronal death. Zinc is released by neurons under several conditions in which microglial activation occurs, and zinc chelators can reduce neuronal death in animal models of cerebral ischemia and neurodegenerative disorders. Here, we show that zinc directly triggers microglial activation. Microglia transfected with a nuclear factor-kappaB (NF-kappaB) reporter gene showed a severalfold increase in NF-kappaB activity in response to 30 microm zinc. Cultured mouse microglia exposed to 15-30 microm zinc increased nitric oxide production, increased F4/80 expression, altered cytokine expression, and assumed the activated morphology. Zinc-induced microglial activation was blocked by inhibiting NADPH oxidase, poly(ADP-ribose) polymerase-1 (PARP-1), or NF-kappaB activation. Zinc injected directly into mouse brain induced microglial activation in wild-type mice, but not in mice genetically lacking PARP-1 or NADPH oxidase activity. Endogenous zinc release, induced by cerebral ischemia reperfusion, likewise induced a robust microglial reaction, and this reaction was suppressed by the zinc chelator CaEDTA. Together, these results suggest that extracellular zinc triggers microglial activation through the sequential activation of NADPH oxidase, PARP-1, and NF-kappaB. These findings identify a novel trigger for microglial activation and a previously unrecognized mechanism by which zinc may contribute to neurological disorders. PMID- 18509045 TI - Activation of TRPV1 contributes to morphine tolerance: involvement of the mitogen activated protein kinase signaling pathway. AB - Tolerance to the analgesic effects of opioids occurs after their chronic administration, a pharmacological phenomenon that has been associated with the development of abnormal pain sensitivity such as hyperalgesia. In the present study, we investigated the role of TRPV1, which is crucial for the transduction of noxious chemical and thermal stimuli, in morphine tolerance and tolerance associated thermal hyperalgesia. After chronic morphine treatment, a marked increase in TRPV1 immunoreactivity (IR) was detected in L4 dorsal root ganglion (DRG) neurons, spinal cord dorsal horn, and sciatic nerve. Real-time reverse transcription (RT)-PCR demonstrated that TRPV1 mRNA was upregulated in spinal cord and sciatic nerve but not in the DRG. Intrathecal pretreatment with SB366791 [N-(3-methoxyphenyl)-4-chlorocinnamide], a selective antagonist of TRPV1, attenuated both morphine tolerance and associated thermal hyperalgesia. Chronic morphine exposure induced increases in phosphorylation of mitogen-activated protein kinases (MAPKs), including p38 MAPK-IR, extracellular signal-regulated protein kinase (ERK)-IR, and c-Jun N-terminal kinase (JNK)-IR, in L4 DRG neurons. Intrathecal administration of the selective p38, ERK, or JNK inhibitors not only reduced morphine tolerance and associated thermal hyperalgesia but also suppressed the morphine-induced increase of TRPV1-IR in DRG neurons, spinal cord, and sciatic nerve and of mRNA levels in spinal cord and sciatic nerve. Together, we have identified a novel mechanism by which sustained morphine treatment results in tolerance and tolerance-associated thermal hyperalgesia, by regulating TRPV1 expression, in a MAPK-dependent manner. Thus, blocking TRPV1 might be a way to reduce morphine tolerance. PMID- 18509046 TI - The h channel mediates location dependence and plasticity of intrinsic phase response in rat hippocampal neurons. AB - The presence of phenomenological inductances in neuronal membrane has been known for more than one-half a century. Despite this, the dramatic contributions of such inductive elements to the amplitude and, especially, phase of neuronal impedance, and their roles in modulating temporal dynamics of neuronal responses have surprisingly remained unexplored. In this study, we demonstrate that the h channel contributes a location-dependent and plastic phenomenological inductive component to the input impedance of CA1 pyramidal neurons. Specifically, we show that the h channels introduce an apparent negative delay in the local voltage response of these neurons with respect to the injected current within the theta frequency range. The frequency range and the extent of this lead expand with increases in h current either through hyperpolarization, or with increasing distance of dendritic location from the soma. We also demonstrate that a spatially widespread increase in this inductive phase component accompanies long term potentiation. Finally, using impedance analysis, we show that both location and activity dependence of intrinsic phase response are attributable not to changes in a capacitive or a leak component, but to changes in h-channel properties. Our results suggest that certain voltage-gated ion channels can differentially regulate internal time delays within neurons, thus providing them with an independent control mechanism in temporal coding of neuronal information. Our analyses and results also establish impedance as a powerful measure of intrinsic dynamics and excitability, given that it quantifies temporal relationships among signals and excitability as functions of input frequency. PMID- 18509047 TI - The neural mechanisms underlying the influence of pavlovian cues on human decision making. AB - In outcome-specific transfer, pavlovian cues that are predictive of specific outcomes bias action choice toward actions associated with those outcomes. This transfer occurs despite no explicit training of the instrumental actions in the presence of pavlovian cues. The neural substrates of this effect in humans are unknown. To address this, we scanned 23 human subjects with functional magnetic resonance imaging while they made choices between different liquid food rewards in the presence of pavlovian cues previously associated with one of these outcomes. We found behavioral evidence of outcome-specific transfer effects in our subjects, as well as differential blood oxygenation level-dependent activity in a region of ventrolateral putamen when subjects chose, respectively, actions consistent and inconsistent with the pavlovian-predicted outcome. Our results suggest that choosing an action incompatible with a pavlovian-predicted outcome might require the inhibition of feasible but nonselected action-outcome associations. The results of this study are relevant for understanding how marketing actions can affect consumer choice behavior as well as for how environmental cues can influence drug-seeking behavior in addiction. PMID- 18509048 TI - T cell receptor signaling controls Foxp3 expression via PI3K, Akt, and mTOR. AB - Regulatory T (Treg) cells safeguard against autoimmunity and immune pathology. Because determinants of the Treg cell fate are not completely understood, we have delineated signaling events that control the de novo expression of Foxp3 in naive peripheral CD4 T cells and in thymocytes. We report that premature termination of TCR signaling and inibition of phosphatidyl inositol 3-kinase (PI3K) p110alpha, p110delta, protein kinase B (Akt), or mammalian target of rapamycin (mTOR) conferred Foxp3 expression and Treg-like gene expression profiles. Conversely, continued TCR signaling and constitutive PI3K/Akt/mTOR activity antagonised Foxp3 induction. At the chromatin level, di- and trimethylation of lysine 4 of histone H3 (H3K4me2 and -3) near the Foxp3 transcription start site (TSS) and within the 5' untranslated region (UTR) preceded active Foxp3 expression and, like Foxp3 inducibility, was lost upon continued TCR stimulation. These data demonstrate that the PI3K/Akt/mTOR signaling network regulates Foxp3 expression. PMID- 18509049 TI - Electrostatic basis for the unidirectionality of the primary proton transfer in cytochrome c oxidase. AB - Gaining detailed understanding of the energetics of the proton-pumping process in cytochrome c oxidase (CcO) is one of the challenges of modern biophysics. Despite promising mechanistic proposals, most works have not related the activation barriers of the different assumed steps to the protein structure, and there has not been a physically consistent model that reproduced the barriers needed to create a working pump. This work reevaluates the activation barriers for the primary proton transfer (PT) steps by calculations that reflect all relevant free energy contributions, including the electrostatic energies of the generated charges, the energies of water insertion, and large structural rearrangements of the donor and acceptor. The calculations have reproduced barriers that account for the directionality and sequence of events in the primary PT in CcO. It has also been found that the PT from Glu-286 (E) to the propionate of heme a(3) (Prd) provides a gate for an initial back leakage from the high pH side of the membrane. Interestingly, the rotation of E that brings it closer to Prd appears to provide a way for blocking competing pathways in the primary PT. Our study elucidates and quantifies the nature of the control of the directionality in the primary PT in CcO and provides instructive insight into the role of the water molecules in biological PT, showing that "bridges" of several water molecules in hydrophobic regions present a problem (rather than a solution) that is minimized in the primary PT. PMID- 18509050 TI - Load-dependent ADP binding to myosins V and VI: implications for subunit coordination and function. AB - Dimeric myosins V and VI travel long distances in opposite directions along actin filaments in cells, taking multiple steps in a "hand-over-hand" fashion. The catalytic cycles of both myosins are limited by ADP dissociation, which is considered a key step in the walking mechanism of these motors. Here, we demonstrate that external loads applied to individual actomyosin V or VI bonds asymmetrically affect ADP affinity, such that ADP binds weaker under loads assisting motility. Model-based analysis reveals that forward and backward loads modulate the kinetics of ADP binding to both myosins, although the effect is less pronounced for myosin VI. ADP dissociation is modestly accelerated by forward loads and inhibited by backward loads. Loads applied in either direction slow ADP binding to myosin V but accelerate binding to myosin VI. We calculate that the intramolecular load generated during processive stepping is approximately 2 pN for both myosin V and myosin VI. The distinct load dependence of ADP binding allows these motors to perform different cellular functions. PMID- 18509051 TI - Immunity to a salivary protein of a sand fly vector protects against the fatal outcome of visceral leishmaniasis in a hamster model. AB - Visceral leishmaniasis (VL) is a fatal disease for humans, and no vaccine is currently available. Sand fly salivary proteins have been associated with protection against cutaneous leishmaniasis. To test whether vector salivary proteins can protect against VL, a hamster model was developed involving intradermal inoculation in the ears of 100,000 Leishmania infantum chagasi parasites together with Lutzomyia longipalpis saliva to mimic natural transmission by sand flies. Hamsters developed classical signs of VL rapidly, culminating in a fatal outcome 5-6 months postinfection. Saliva had no effect on the course of infection in this model. Immunization with 16 DNA plasmids coding for salivary proteins of Lu. longipalpis resulted in the identification of LJM19, a novel 11-kDa protein, that protected hamsters against the fatal outcome of VL. LJM19-immunized hamsters maintained a low parasite load that correlated with an overall high IFN-gamma/TGF-beta ratio and inducible NOS expression in the spleen and liver up to 5 months postinfection. Importantly, a delayed-type hypersensitivity response with high expression of IFN-gamma was also noted in the skin of LJM19-immunized hamsters 48 h after exposure to uninfected sand fly bites. Induction of IFN-gamma at the site of bite could partly explain the protection observed in the viscera of LJM19-immunized hamsters through direct parasite killing and/or priming of anti-Leishmania immunity. We have shown that immunity to a defined salivary protein (LJM19) confers powerful protection against the fatal outcome of a parasitic disease, which reinforces the concept of using components of arthropod saliva in vaccine strategies against vector-borne diseases. PMID- 18509052 TI - Controlling the efficiency of an artificial light-harvesting complex. AB - Adaptive femtosecond pulse shaping in an evolutionary learning loop is applied to a bioinspired dyad molecule that closely mimics the early-time photophysics of the light-harvesting complex 2 (LH2) photosynthetic antenna complex. Control over the branching ratio between the two competing pathways for energy flow, internal conversion (IC) and energy transfer (ET), is realized. We show that by pulse shaping it is possible to increase independently the relative yield of both channels, ET and IC. The optimization results are analyzed by using Fourier analysis, which gives direct insight to the mechanism featuring quantum interference of a low-frequency mode. The results from the closed-loop experiments are repeatable and robust and demonstrate the power of coherent control experiments as a spectroscopic tool (i.e., quantum-control spectroscopy) capable of revealing functionally relevant molecular properties that are hidden from conventional techniques. PMID- 18509053 TI - Involvement of estrogen-related receptors in transcriptional response to hypoxia and growth of solid tumors. AB - The development of intratumoral hypoxia is a universal hallmark of rapidly growing solid tumors. Adaptation to the hypoxic environment, which is critical for tumor cell survival and growth, is mediated primarily through a hypoxia inducible factor (HIF)-dependent transcriptional program. HIF activates genes that facilitate crucial adaptive mechanisms including increased glucose uptake and glycolysis and tumor angiogenesis, making it an important therapeutic target. However, the HIF-dependent transcriptional mechanism remains incompletely understood, and targeting HIF is a difficult endeavor. Here, we show that the orphan nuclear receptor estrogen-related receptors (ERRs) physically interact with HIF and stimulate HIF-induced transcription. Importantly, ERRs appear to be essential for HIF's function. Transcriptional activation of hypoxic genes in cells cultured under hypoxia is largely blocked by suppression of ERRs through expression of a dominant negative form of ERR or treatment with a pharmacological ERR inhibitor, diethylstilbestrol. Systematic administration of diethylstilbestrol severely diminished growth and angiogenesis of tumor xenografts in vivo. Because nuclear receptors are outstanding targets for drug discovery, the findings not only may offer mechanistic insights into HIF-mediated transcription but also may open new avenues for targeting the HIF pathway for cancer therapy. PMID- 18509054 TI - Spatial scaling of functional gene diversity across various microbial taxa. AB - Understanding the spatial patterns of organisms and the underlying mechanisms shaping biotic communities is a central goal in community ecology. One of the most well documented spatial patterns in plant and animal communities is the positive-power law relationship between species (or taxa) richness and area. Such taxa-area relationships (TARs) are one of the principal generalizations in ecology, and are fundamental to our understanding of the distribution of global biodiversity. However, TARs remain elusive in microbial communities, especially in soil habitats, because of inadequate sampling methodologies. Here, we describe TARs as gene-area relationships (GARs), at a whole-community level, across various microbial functional and phylogenetic groups in a forest soil, using a comprehensive functional gene array with >24,000 probes. Our analysis indicated that the forest soil microbial community exhibited a relatively flat gene-area relationship (slope z = 0.0624), but the z values varied considerably across different functional and phylogenetic groups (z = 0.0475-0.0959). However, the z values are several times lower than those commonly observed in plants and animals. These results suggest that the turnover in space of microorganisms may be, in general, lower than that of plants and animals. PMID- 18509055 TI - Natural variation in Pristionchus pacificus insect pheromone attraction involves the protein kinase EGL-4. AB - The geographical mosaic theory of coevolution predicts that different local species interactions will shape population traits, but little is known about the molecular factors involved in mediating the specificity of these interactions. Pristionchus nematodes associate with different scarab beetles around the world, with Pristionchus pacificus isolated primarily from the oriental beetle in Japan. In particular, the constituent populations of P. pacificus represent a rare opportunity to study multiple specialized interactions and the mechanisms that influence population traits at the genetic level. We identified a component of the cGMP signaling pathway to be involved in the natural variation for sensing the insect pheromone ETDA, using targeted introgression lines, exogenous cGMP treatment, and a null egl-4 allele. Our data strongly implicate egl-4 as one of several loci involved in behavioral variation in P. pacificus populations. That EGL-4 homologs have been independently implicated for behavioral variations in other invertebrate models suggests that EGL-4 may act as a modulator for interspecies behavioral repertoires across large phylogenetic distances. PMID- 18509056 TI - Passivation of irregular surfaces accessed by diffusion. AB - We investigate the process of progressive passivation of irregular surfaces accessed by diffusion. More precisely, we quantify through numerical simulations how the activity of the von Koch surface is gradually transferred from its initially active (or absorbing) regions to its less accessible regions. We show that in three dimensions, in sharp contrast with the two-dimensional case, the size of the successive active zones steadily decreases during the passivation process, even though a large quantity of alive surface remains available. As a consequence, in three dimensions, the evolution of the efficiency of a surface accessed by diffusion (i.e., by a Laplacian field) can exhibit long-tail behaviors that, unlike in two dimensions, strongly depend on its specific geometry. This fact has important implications for the design of heterogeneous catalysts under deactivation conditions, for the performance of heat exchangers subjected to passivation by "fouling," and for changes in the behavior of the digestive system, where the activity of the absorbing intestinal membrane can be substantially affected by inflammatory disorders. PMID- 18509057 TI - Muscle insulin resistance: a case of fat overconsumption, not mitochondrial dysfunction. PMID- 18509058 TI - Dimeric subunit stoichiometry of the human voltage-dependent proton channel Hv1. AB - In voltage-gated Na(+), K(+), and Ca(2+) channels, four voltage-sensor domains operate on a central pore domain in response to membrane voltage. In contrast, the voltage-gated proton channel (Hv) contains only a voltage-sensor domain, lacking a separate pore domain. The subunit stoichiometry and organization of Hv has been unknown. Here, we show that human Hv1 forms a dimer in the membrane and define regions that are close to the dimer interface by using cysteine cross linking. Two dimeric interfaces appear to exist in Hv1, one mediated by S1 and the adjacent extracellular loop, and the other mediated by a putative intracellular coiled-coil domain. It may be significant that Hv1 uses for its dimer interface a surface that corresponds to the interface between the voltage sensor and pore in Kv channels. PMID- 18509059 TI - A latitudinal diversity gradient in planktonic marine bacteria. AB - For two centuries, biologists have documented a gradient of animal and plant biodiversity from the tropics to the poles but have been unable to agree whether it is controlled primarily by productivity, temperature, or historical factors. Recent reports that find latitudinal diversity gradients to be reduced or absent in some unicellular organisms and attribute this to their high abundance and dispersal capabilities would suggest that bacteria, the smallest and most abundant organisms, should exhibit no latitudinal pattern of diversity. We used amplified ribosomal intergenic spacer analysis (ARISA) whole-assemblage genetic fingerprinting to quantify species richness in 103 near-surface samples of marine bacterial plankton, taken from tropical to polar in both hemispheres. We found a significant latitudinal gradient in richness. The data can help to evaluate hypotheses about the cause of the gradient. The correlations of richness with latitude and temperature were similarly strong, whereas correlations with parameters relating to productivity (chlorophyll, annual primary productivity, bacterial abundance) and other variables (salinity and distance to shore) were much weaker. Despite the high abundance and potentially high dispersal of bacteria, they exhibit geographic patterns of species diversity that are similar to those seen in other organisms. The latitudinal gradient in marine bacteria supports the hypothesis that the kinetics of metabolism, setting the pace for life, has strong influence on diversity. PMID- 18509060 TI - LRP6 transduces a canonical Wnt signal independently of Axin degradation by inhibiting GSK3's phosphorylation of beta-catenin. AB - Wnt/beta-catenin signaling controls various cell fates in metazoan development and is misregulated in several cancers and developmental disorders. Binding of a Wnt ligand to its transmembrane coreceptors inhibits phosphorylation and degradation of the transcriptional coactivator beta-catenin, which then translocates to the nucleus to regulate target gene expression. To understand how Wnt signaling prevents beta-catenin degradation, we focused on the Wnt coreceptor low-density lipoprotein receptor-related protein 6 (LRP6), which is required for signal transduction and is sufficient to activate Wnt signaling when overexpressed. LRP6 has been proposed to stabilize beta-catenin by stimulating degradation of Axin, a scaffold protein required for beta-catenin degradation. In certain systems, however, Wnt-mediated Axin turnover is not detected until after beta-catenin has been stabilized. Thus, LRP6 may also signal through a mechanism distinct from Axin degradation. To establish a biochemically tractable system to test this hypothesis, we expressed and purified the LRP6 intracellular domain from bacteria and show that it promotes beta-catenin stabilization and Axin degradation in Xenopus egg extract. Using an Axin mutant that does not degrade in response to LRP6, we demonstrate that LRP6 can stabilize beta-catenin in the absence of Axin turnover. Through experiments in egg extract and reconstitution with purified proteins, we identify a mechanism whereby LRP6 stabilizes beta catenin independently of Axin degradation by directly inhibiting GSK3's phosphorylation of beta-catenin. PMID- 18509061 TI - Control of endothelial cell proliferation and migration by VEGF signaling to histone deacetylase 7. AB - VEGF has been shown to regulate endothelial cell (EC) proliferation and migration. However, the nuclear mediators of the actions of VEGF in ECs have not been fully defined. We show that VEGF induces the phosphorylation of three conserved serine residues in histone deacetylase 7 (HDAC7) via protein kinase D, which promotes nuclear export of HDAC7 and activation of VEGF-responsive genes in ECs. Expression of a signal-resistant HDAC7 mutant protein in ECs inhibits proliferation and migration in response to VEGF. These results demonstrate that phosphorylation of HDAC7 serves as a molecular switch to mediate VEGF signaling and endothelial function. PMID- 18509062 TI - Cidea is associated with lipid droplets and insulin sensitivity in humans. AB - Storage of energy as triglyceride in large adipose-specific lipid droplets is a fundamental need in all mammals. Efficient sequestration of fat in adipocytes also prevents fatty acid overload in skeletal muscle and liver, which can impair insulin signaling. Here we report that the Cide domain-containing protein Cidea, previously thought to be a mitochondrial protein, colocalizes around lipid droplets with perilipin, a regulator of lipolysis. Cidea-GFP greatly enhances lipid droplet size when ectopically expressed in preadipocytes or COS cells. These results explain previous findings showing that depletion of Cidea with RNAi markedly elevates lipolysis in human adipocytes. Like perilipin, Cidea and the related lipid droplet protein Cidec/FSP27 are controlled by peroxisome proliferator-activated receptor gamma (PPARgamma). Treatment of lean or obese mice with the PPARgamma agonist rosiglitazone markedly up-regulates Cidea expression in white adipose tissue (WAT), increasing lipid deposition. Strikingly, in both omental and s.c. WAT from BMI-matched obese humans, expression of Cidea, Cidec/FSP27, and perilipin correlates positively with insulin sensitivity (HOMA-IR index). Thus, Cidea and other lipid droplet proteins define a novel, highly regulated pathway of triglyceride deposition in human WAT. The data support a model whereby failure of this pathway results in ectopic lipid accumulation, insulin resistance, and its associated comorbidities in humans. PMID- 18509063 TI - High-fat diets cause insulin resistance despite an increase in muscle mitochondria. AB - It has been hypothesized that insulin resistance is mediated by a deficiency of mitochondria in skeletal muscle. In keeping with this hypothesis, high-fat diets that cause insulin resistance have been reported to result in a decrease in muscle mitochondria. In contrast, we found that feeding rats high-fat diets that cause muscle insulin resistance results in a concomitant gradual increase in muscle mitochondria. This adaptation appears to be mediated by activation of peroxisome proliferator-activated receptor (PPAR)delta by fatty acids, which results in a gradual, posttranscriptionally regulated increase in PPAR gamma coactivator 1alpha (PGC-1alpha) protein expression. Similarly, overexpression of PPARdelta results in a large increase in PGC-1alpha protein in the absence of any increase in PGC-1alpha mRNA. We interpret our findings as evidence that raising free fatty acids results in an increase in mitochondria by activating PPARdelta, which mediates a posttranscriptional increase in PGC-1alpha. Our findings argue against the concept that insulin resistance is mediated by a deficiency of muscle mitochondria. PMID- 18509064 TI - Building evaluation capacity in Spain: a case study of rural development and empowerment in the European Union. AB - The development of European Community administrative authority has greatly influenced the development of an evaluation culture among the southern and central member states of the European Union. The present case study from Spain provides an example of this diffusion through the use of an empowerment evaluation approach to build evaluation capacity within the context of rural development. The study focuses on the evaluation process over a 10-year period of three local development programs run by three local partnerships created in the rural areas of the Madrid region through the European LEADER initiative. Critical components of empowerment evaluation are discussed, as well as the impact, limitations, difficulties, and applicability of the approach to rural development in the European Union. Conclusions are presented concerning evaluation capacity building and the development of an evaluation culture. PMID- 18509065 TI - Overcoming S-phase checkpoint-mediated resistance: sequence-dependent synergy of gemcitabine and 7-ethyl-10-hydroxycamptothecin (SN-38) in human carcinoma cell lines. AB - Although agents that inhibit DNA synthesis are widely used in the treatment of cancer, the optimal method for combining such agents and the mechanism of their synergy is poorly understood. The present study examined the effects of combining gemcitabine (2',2'-difluoro 2'-deoxycytidine) and 7-ethyl-10-hydroxycamptothecin (SN-38; the active metabolite of irinotecan), two S-phaseselective agents that individually have broad antitumor activity, in human cancer cells in vitro. Colony-forming assays revealed that simultaneous treatment of Ovcar-5 ovarian cancer cells or BxPC-3 pancreatic cancer cells with gemcitabine and SN-38 resulted in antagonistic effects. In contrast, sequential treatment with these two agents in either order resulted in synergistic anti-proliferative effects, although the mechanism of synergy varied with the sequence. In particular, SN-38 arrested cells in S phase, enhanced the accumulation of gemcitabine metabolites, and diminished checkpoint kinase 1, thereby sensitizing cells in the SN-38 --> gemcitabine sequence. Gemcitabine treatment followed by removal allowed prolonged progression through S phase, contributing to synergy of the gemcitabine --> SN-38 sequence. These results collectively suggest that S-phase-selective agents might exhibit more cytotoxicity when administered sequentially rather than simultaneously. PMID- 18509066 TI - Mutational analysis of the conserved Asp2.50 and ERY motif reveals signaling bias of the urotensin II receptor. AB - Class A (rhodopsin-like) G protein-coupled receptors possess conserved residues and motifs that are important for their specific activity. In the present study, we examined the role of residue Asp97(2.50) as well as residues Glu147(3.49), Arg148(3.50), and Tyr149(3.51) of the ERY motif on the functionality of the urotensin II receptor (UT). Mutations D97(2.50)A, R148(3.50)A, and R148(3.50)H abolished the ability of UT to activate phospholipase C, whereas mutations E147(3.49)A and Y149(3.51)A reduced the ability to activate PLC by 50%. None of the mutants exhibited constitutive activity. However, R148(3.50)A and R148(3.50)H promoted ERK1/2 activation, which was abolished by 4-(3-chloroanilino)-6,7 dimethoxyquinazoline (AG1478), an inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase activity. Both these mutants were capable of directly activating EGFR, which confirmed that they activated the mitogen-activated protein kinase (MAPK) pathway by a Galpha(q/11)-independent transactivation of EGFR. The D97(2.50)A, R148(3.50)A, and R148(3.50)H mutants did not readily internalize and did not promote translocation or colocalize with beta-arrestin2 GFP. Finally, the agonist-induced internalization of the E147(3.49)A mutant receptor was significantly increased compared with wild-type receptor. This study highlights the major contribution of the conserved Asp(2.50) residue to the functionality of the UT receptor. The Arg residue in the ERY motif of UT is an important structural element in signaling crossroads that determine whether Galpha(q/11)-dependent and -independent events can occur. PMID- 18509067 TI - Effect of pregnancy on cytochrome P450 3a and P-glycoprotein expression and activity in the mouse: mechanisms, tissue specificity, and time course. AB - The plasma concentrations of orally administered anti-human immunodeficiency virus protease inhibitors are significantly reduced during human and mouse pregnancy. We have shown that in the mouse, at gestational day 19, this reduction is due to increased hepatic cytochrome P450 3a (Cyp3a) protein expression and activity. In the current study, we investigated the mechanisms by which Cyp3a activity is increased by pregnancy and the time course of change in expression of Cyp3a and P-glycoprotein (P-gp) in various tissues. We found that hepatic transcripts of Cyp3a16, Cyp3a41, and Cyp3a44 were significantly increased during pregnancy, whereas those of Cyp3a11 and Cyp3a25 were significantly decreased. This resulted in a net increase in Cyp3a protein expression and activity in the liver during pregnancy. The increase in Cyp3a41 and Cyp3a44 transcripts was positively correlated (p < 0.05) with hepatocyte nuclear factor 6 and estrogen receptor-alpha transcripts. The pregnancy-related factors that transcriptionally activated mouse Cyp3a isoforms also activated the human CYP3A4 promoter in pregnant CYP3A4-promoter-luciferase transgenic (CYP3A4-tg) mice. In contrast, intestinal Cyp3a protein expression was not significantly affected by pregnancy. No change in P-gp protein expression was observed in the liver or kidney during pregnancy, although a significant decrease was observed in the placenta. Because hepatic CYP3A activity also seems to be induced during human pregnancy, the mouse (including CYP3A4-tg mouse) seems to be an excellent animal model to determine the molecular mechanisms for such an induction. PMID- 18509068 TI - Resting states affect spontaneous BOLD oscillations in sensory and paralimbic cortex. AB - The brain exhibits spontaneous neural activity that depends on the behavioral state of the organism. We asked whether the blood oxygenation level-dependent (BOLD) signal reflects these modulations. BOLD was measured under three steady state conditions: while subjects kept their eyes closed, kept their eyes open, or while fixating. The BOLD spectral density was calculated across brain voxels and subjects. Visual, sensory-motor, auditory, and retrosplenial cortex showed modulations of the BOLD spectral density by resting state type. All modulated regions showed greater spontaneous BOLD oscillations in the eyes closed than the eyes open or fixation conditions, suggesting that the differences were endogenously driven. Next, we examined the pattern of correlations between regions whose ongoing BOLD signal was modulated by resting state type. Regional neuronal correlations were estimated using an analytic procedure from the comparison of BOLD-BOLD covariances in the fixation and eyes closed conditions. Most regions were highly correlated with one another, with the exception of the primary visual cortices, which showed low correlations with the other regions. In conclusion, changes in resting state were associated with synchronous modulations of spontaneous BOLD oscillations in cortical sensory areas driven by two spatially overlapping, but temporally uncorrelated signals. PMID- 18509069 TI - Noninvasive stimulation of human corticospinal axons innervating leg muscles. AB - These studies investigated whether a single electrical stimulus over the thoracic spine activates corticospinal axons projecting to human leg muscles. Transcranial magnetic stimulation of the motor cortex and electrical stimulation over the thoracic spine were paired at seven interstimulus intervals, and surface electromyographic responses were recorded from rectus femoris, tibialis anterior, and soleus. The interstimulus intervals (ISIs) were set so that the first descending volley evoked by cortical stimulation had not arrived at (positive ISIs), was at the same level as (0 ISI) or had passed (negative ISIs) the site of activation of descending axons by the thoracic stimulation at the moment of its delivery. Compared with the responses to motor cortical stimulation alone, responses to paired stimuli were larger at negative ISIs but reduced at positive ISIs in all three leg muscles. This depression of responses at positive ISIs is consistent with an occlusive interaction in which an antidromic volley evoked by the thoracic stimulation collides with descending volleys evoked by cortical stimulation. The cortical and spinal stimuli activate some of the same corticospinal axons. Thus it is possible to examine the excitability of lower limb motoneuron pools to corticospinal inputs without the confounding effects of changes occurring within the motor cortex. PMID- 18509070 TI - Silencing-induced metaplasticity in hippocampal cultured neurons. AB - Silencing-induced homeostatic plasticity is usually expressed as a change in the amplitude or the frequency of miniature postsynaptic currents. Here we report that, prolonged (approximately 24 h) silencing of mature (20-22 days in vitro) cultured hippocampal neurons using the voltage-gated sodium channel blocker tetrodotoxin (TTX) produced no effects on the amplitude or frequency of the miniature excitatory postsynaptic currents (mEPSCs). However, the silencing changed the intrinsic membrane properties of the neurons, resulting in an increased excitability and rate of action potentials firing upon TTX washout. Allowing neurons to recover in TTX-free recording solution for a short period of time after the silencing resulted in potentiation of mEPSC amplitudes. This form of activity-dependent potentiation is different from classical long-term potentiation, as similar potentiation was not seen in nonsilenced neurons treated with bicuculline to raise their spiking activity to the same level displayed by the silenced neurons during TTX washout. Also, the potentiation of mEPSC amplitudes after the recovery period was not affected by the N-methyl-d-aspartate receptor blocker d-2-amino-5-phosponopentanoic acid or by the calcium/calmodulin dependent kinase II (CaMKII) inhibitor KN-62 but was abolished by the L-type calcium channel blocker nifedipine. We thus conclude that the potentiation of mEPSC amplitudes following brief recovery of spiking activity in chronically silenced neurons represents a novel form of metaplasticity that differs from the conventional models of homeostatic synaptic plasticity. PMID- 18509071 TI - Variability reduction in interaural time difference tuning in the barn owl. AB - The interaural time difference (ITD) is the primary auditory cue used by the barn owl for localization in the horizontal direction. ITD is initially computed by circuits consisting of axonal delay lines from one of the cochlear nuclei and coincidence detector neurons in the nucleus laminaris (NL). NL projects directly to the anterior part of the dorsal lateral lemniscal nucleus (LLDa), and this area projects to the core of the central nucleus of the inferior colliculus (ICcc) in the midbrain. To show the selectivity of an NL neuron for ITD requires averaging of responses over several stimulus presentations for each ITD. In contrast, ICcc neurons detect their preferred ITD in a single burst of stimulus. We recorded extracellularly the responses of LLDa neurons to ITD in anesthetized barn owls and show that this ability is already present in LLDa, raising the possibility that ICcc inherits its noise reduction property from LLDa. PMID- 18509072 TI - Asymmetric recovery in cerebellar-deficient mice following unilateral labyrinthectomy. AB - The term "vestibular compensation" refers to the resolution of motor deficits resulting from a peripheral vestibular lesion. We investigated the role of the cerebellum in the compensation process by characterizing the vestibuloocular reflex (VOR) evoked by head rotations at frequencies and velocities similar to those in natural behaviors in wild-type (WT) versus cerebellar-deficient Lurcher (Lc/+) mice. We found that during exploratory activity, normal mice produce head rotations largely consisting of frequencies < or =4 Hz and velocities and accelerations as large as 400 degrees/s and 5,000 degrees/s2, respectively. Accordingly, the VOR was characterized using sinusoidal rotations (0.2-4 Hz) as well as transient impulses (approximately 400 degrees/s; approximately 2,000 degrees/s2). Before lesions, WT and Lc/+ mice produced similar VOR responses to sinusoidal rotation. Lc/+ mice, however, had significantly reduced gains for transient stimuli. After unilateral labyrinthectomy, VOR recovery followed a similar course for WT and Lc/+ groups during the first week: gain was reduced by 80% for ipsilesionally directed head rotations on day 1 and improved for both strains to values of approximately 0.4 by day 5. Moreover, responses evoked by contralesionally directed rotations returned to prelesion in both strains within this period. However, unlike WT, which showed improving responses to ipsilesionally directed rotations, recovery plateaued after first week for Lc/+ mice. Our results show that despite nearly normal recovery in the acute phase, long-term compensation is compromised in Lc/+. We conclude that cerebellar pathways are critical for long-term restoration of VOR during head rotation toward the lesioned side, while noncerebellar pathways are sufficient to restore proper gaze stabilization during contralesionally directed movements. PMID- 18509073 TI - Temporal processing across multiple topographic maps in the electrosensory system. AB - Multiple topographic representations of sensory space are common in the nervous system and presumably allow organisms to separately process particular features of incoming sensory stimuli that vary widely in their attributes. We compared the response properties of sensory neurons within three maps of the body surface that are arranged strictly in parallel to two classes of stimuli that mimic prey and conspecifics, respectively. We used information-theoretic approaches and measures of phase locking to quantify neuronal responses. Our results show that frequency tuning in one of the three maps does not depend on stimulus class. This map acts as a low-pass filter under both conditions. A previously described stimulus-class dependent switch in frequency tuning is shown to occur in the other two maps. Only a fraction of the information encoded by all neurons could be recovered through a linear decoder. Particularly striking were low-pass neurons the information of which in the high-frequency range could not be decoded linearly. We then explored whether intrinsic cellular mechanisms could partially account for the differences in frequency tuning across maps. Injection of a Ca2+ chelator had no effect in the map with low-pass characteristics. However, injection of the same Ca2+ chelator in the other two maps switched the tuning of neurons from band pass/high-pass to low-pass. These results show that Ca2+-dependent processes play an important part in determining the functional roles of different sensory maps and thus shed light on the evolution of this important feature of the vertebrate brain. PMID- 18509074 TI - Hyperpolarization-activated cyclic nucleotide-gated channels in mouse vomeronasal sensory neurons. AB - Hyperpolarization-activated currents (Ih) are present in several neurons of the central and peripheral nervous system. However, Ih in neurons of the vomeronasal organ (VNO) is not well characterized. We studied the properties of Ih in sensory neurons from acute slices of mouse VNO. In voltage-clamp studies, Ih was identified by the characteristic kinetics of activation, voltage dependence, and blockage by Cs+ or ZD-7288, two blockers of the Ih. Forskolin, an activator of adenylyl cyclase, shifted the activation curve for Ih to less negative potentials. A comparison of Ih properties in VNO neurons with those of heterologously expressed hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, together with RT-PCR experiments in VNO, indicate that Ih is caused by HCN2 and/or HCN4 subunits. In current-clamp recordings, blocking Ih with ZD-7288 induced a hyperpolarization of 5.1 mV, an increase in input resistance, a decrease in the sensitivity to elicit action potentials in response to small current injections, and did not modify the frequency of action potentials elicited by a large current injection. It has been shown that in VNO neurons some pheromones induce a decrease in cAMP concentration, but the physiological role of cAMP is unknown. After application of blockers of adenylyl cyclase, we measured a hyperpolarization of 5.1 mV in 11 of 14 neurons, suggesting that basal levels of cAMP could modulate the resting potential. In conclusion, these results show that mouse VNO neurons express HCN2 and/or HCN4 subunits and that Ih contributes to setting the resting membrane potential and to increase excitability at stimulus threshold. PMID- 18509075 TI - The activity of spinal commissural interneurons during fictive locomotion in the lamprey. AB - Commissural interneurons in the lamprey coordinate activity of the hemisegmental oscillators to ensure proper left-right alternation during swimming. The activity of interneuronal axons at the ventral commissure was studied together with potential target motoneurons during fictive locomotion in the isolated lamprey spinal cord. To estimate the unperturbed activity of the interneurons, axonal recordings were chosen because soma recordings inevitably will affect the level of membrane depolarization and thereby spike initiation. Of 227 commissural axons recorded during locomotor activity, 14 produced inhibitory and 3 produced excitatory postsynaptic potentials (PSPs) in target motoneurons. The axons typically fired multiple spikes per locomotor cycle, with approximately 10 Hz sustained frequency. The average shortest spike interval in a burst corresponded to an instantaneous frequency of approximately 50 Hz for both the excitatory and inhibitory axons. The maximum number of spikes per locomotor cycle was inversely related to the locomotor frequency, in accordance with previous observations in the spinal hemicord preparation. In axons that fired multiple spikes per cycle, the mean interspike intervals were in the range in which the amplitude of the slow afterhyperpolarization (sAHP) is large, providing further support for the role of the sAHP in spike timing. One hundred ninety-five axons (86%) fired rhythmically during fictive locomotion, with preferred phase of firing distributed over either the segmental locomotor burst phase (40% of axons) or the transitional phase (between bursts; 60%). Thus in lamprey commissural interneurons, we found a broad distribution of firing rates and phases during fictive locomotion. PMID- 18509076 TI - Tonically active inhibition selectively controls feedforward circuits in mouse barrel cortex. AB - Tonic inhibition mediated by extrasynaptic gamma-aminobutyric acid type A (GABA A) receptors is a powerful conductance that controls cell excitability. Throughout the CNS, tonic inhibition is expressed at varying degrees across different cell types. Despite a rich history of cortical interneuron diversity, little is known about tonic inhibition in the different classes of cells in the cerebral cortex. We therefore examined the cell-type specificity and functional significance of tonic inhibition in layer 4 of the mouse somatosensory barrel cortex. In situ hybridization and immunocytochemistry showed moderate delta subunit expression across the barrel structures. Whole cell patch-clamp recordings additionally indicated that significant levels of tonic inhibition can be found across cell types, with differences in the magnitude of inhibition between cell types. To activate tonic currents, we used 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP, a superagonist at delta-subunit containing GABA A receptors) at a concentration that did not affect synaptic decay kinetics. THIP produced greater shifts in baseline holding current in inhibitory cells (low-threshold spiking [LTS], 109 +/- 17 pA; fast spiking [FS], 111 +/- 15 pA) than in excitatory cells (39 +/- 10 pA; P < 0.001). In addition to these differences across cell types, there was also variability within inhibitory cells. FS cells with faster action potentials had larger baseline shifts. Because FS cells are known mediators of feedforward inhibition, we tested whether THIP induced tonic conductance selectively controls feedforward circuits. THIP application resulted in the abolishment of the inhibitory postsynaptic potential in thalamic-evoked disynaptic responses in a subset of excitatory neurons. These data suggest multiple feedforward circuits can be differentiated by the inhibitory control of the presynaptic inhibitory neuron. PMID- 18509078 TI - A biophysical model of synaptic plasticity and metaplasticity can account for the dynamics of the backward shift of hippocampal place fields. AB - Hippocampal place cells in the rat undergo experience-dependent changes when the rat runs stereotyped routes. One such change, the backward shift of the place field center of mass, has been linked by previous modeling efforts to spike timing-dependent plasticity (STDP). However, these models did not account for the termination of the place field shift and they were based on an abstract implementation of STDP that ignores many of the features found in cortical plasticity. Here, instead of the abstract STDP model, we use a calcium-dependent plasticity (CaDP) learning rule that can account for many of the observed properties of cortical plasticity. We use the CaDP learning rule in combination with a model of metaplasticity to simulate place field dynamics. Without any major changes to the parameters of the original model, the present simulations account both for the initial rapid place field shift and for the subsequent slowing down of this shift. These results suggest that the CaDP model captures the essence of a general cortical mechanism of synaptic plasticity, which may underlie numerous forms of synaptic plasticity observed both in vivo and in vitro. PMID- 18509077 TI - Dorsal root ganglion neurons innervating skeletal muscle respond to physiological combinations of protons, ATP, and lactate mediated by ASIC, P2X, and TRPV1. AB - The adequate stimuli and molecular receptors for muscle metaboreceptors and nociceptors are still under investigation. We used calcium imaging of cultured primary sensory dorsal root ganglion (DRG) neurons from C57Bl/6 mice to determine candidates for metabolites that could be the adequate stimuli and receptors that could detect these stimuli. Retrograde DiI labeling determined that some of these neurons innervated skeletal muscle. We found that combinations of protons, ATP, and lactate were much more effective than individually applied compounds for activating rapid calcium increases in muscle-innervating dorsal root ganglion neurons. Antagonists for P2X, ASIC, and TRPV1 receptors suggested that these three receptors act together to detect protons, ATP, and lactate when presented together in physiologically relevant concentrations. Two populations of muscle innervating DRG neurons were found. One responded to low metabolite levels (likely nonnoxious) and used ASIC3, P2X5, and TRPV1 as molecular receptors to detect these metabolites. The other responded to high levels of metabolites (likely noxious) and used ASIC3, P2X4, and TRPV1 as their molecular receptors. We conclude that a combination of ASIC, P2X5 and/or P2X4, and TRPV1 are the molecular receptors used to detect metabolites by muscle-innervating sensory neurons. We further conclude that the adequate stimuli for muscle metaboreceptors and nociceptors are combinations of protons, ATP, and lactate. PMID- 18509079 TI - Active learning: learning a motor skill without a coach. AB - When we learn a new skill (e.g., golf) without a coach, we are "active learners": we have to choose the specific components of the task on which to train (e.g., iron, driver, putter, etc.). What guides our selection of the training sequence? How do choices that people make compare with choices made by machine learning algorithms that attempt to optimize performance? We asked subjects to learn the novel dynamics of a robotic tool while moving it in four directions. They were instructed to choose their practice directions to maximize their performance in subsequent tests. We found that their choices were strongly influenced by motor errors: subjects tended to immediately repeat an action if that action had produced a large error. This strategy was correlated with better performance on test trials. However, even when participants performed perfectly on a movement, they did not avoid repeating that movement. The probability of repeating an action did not drop below chance even when no errors were observed. This behavior led to suboptimal performance. It also violated a strong prediction of current machine learning algorithms, which solve the active learning problem by choosing a training sequence that will maximally reduce the learner's uncertainty about the task. While we show that these algorithms do not provide an adequate description of human behavior, our results suggest ways to improve human motor learning by helping people choose an optimal training sequence. PMID- 18509080 TI - Arousal facilitates collision avoidance mediated by a looming sensitive visual neuron in a flying locust. AB - Locusts have two large collision-detecting neurons, the descending contralateral movement detectors (DCMDs) that signal object approach and trigger evasive glides during flight. We sought to investigate whether vision for action, when the locust is in an aroused state rather than a passive viewer, significantly alters visual processing in this collision-detecting pathway. To do this we used two different approaches to determine how the arousal state of a locust affects the prolonged periods of high-frequency spikes typical of the DCMD response to approaching objects that trigger evasive glides. First, we manipulated arousal state in the locust by applying a brief mechanical stimulation to the hind leg; this type of change of state occurs when gregarious locusts accumulate in high density swarms. Second, we examined DCMD responses during flight because flight produces a heightened physiological state of arousal in locusts. When arousal was induced by either method we found that the DCMD response recovered from a previously habituated state; that it followed object motion throughout approach; and--most important--that it was significantly more likely to generate the maintained spike frequencies capable of evoking gliding dives even with extremely short intervals (1.8 s) between approaches. Overall, tethered flying locusts responded to 41% of simulated approaching objects (sets of 6 with 1.8 s ISI). When we injected epinastine, the neuronal octopamine receptor antagonist, into the hemolymph responsiveness declined to 12%, suggesting that octopamine plays a significant role in maintaining responsiveness of the DCMD and the locust to visual stimuli during flight. PMID- 18509081 TI - V1 response timing and surface filling-in. AB - There is ample evidence from demonstrations such as color induction and stabilized images that information from surface boundaries plays a special role in determining the perception of surface interiors. Surface interiors appear to "fill-in." Psychophysical experiments also show that surface perception involves a slow scale-dependent process distinct from mechanisms involved in contour perception. The present experiments aimed to test the hypothesis that surface perception is associated with relatively slow scale-dependent neural filling-in. We found that responses in macaque primary visual cortex (V1) are slower to surface interiors than responses to optimal bar stimuli. Moreover, we found that the response to a surface interior is delayed relative to the response to the surface's border and the extent of the delay is proportional to the distance between a receptive field and the border. These findings are consistent with some forms of neural filling-in and suggest that V1 may provide the neural substrate for perceptual filling-in. PMID- 18509082 TI - Trunk sensorimotor cortex is essential for autonomous weight-supported locomotion in adult rats spinalized as P1/P2 neonates. AB - Unlike adult spinalized rats, approximately 20% of rats spinalized as postnatal day 1 or 2 (P1/P2) neonates achieve autonomous hindlimb weight support. Cortical representations of mid/low trunk occur only in such rats with high weight support. However, the importance of hindlimb/trunk motor cortex in function of spinalized rats remains unclear. We tested the importance of trunk sensorimotor cortex in their locomotion using lesions guided by cortical microstimulation in P1/P2 weight-supporting neonatal spinalized rats and controls. In four intact control rats, lesions of hindlimb/trunk cortex caused no treadmill deficits. All spinalized rats lesioned in trunk cortex (n = 16: 4 transplant, 6 transect, 6 transect + fibrin glue) lost an average of about 40% of their weight support. Intact trunk cortex was essential to their level of function. Lesion of trunk cortex substantially increased roll of the hindquarters, which correlated to diminished weight support, but other kinematic stepping parameters showed little change. Embryonic day 14 (E14) transplants support development of the trunk motor representations in their normal location. We tested the role of novel relay circuits arising from the grafts in such cortical representations in E14 transplants using the rats that received (noncellular) fibrin glue grafting at P1/P2 (8 allografts and 32 xenografts). Fibrin-repaired rats with autonomous weight support also had trunk cortical representations similar to those of E14 transplant rats. Thus acellular repair and intrinsic plasticity were sufficient to support the observed features. Our data show that effective cortical mechanisms for trunk control are essential for autonomous weight support in P1/P2 spinalized rats and these can be achieved by intrinsic plasticity. PMID- 18509083 TI - Endoplasmic reticulum HSP90b1 (gp96, grp94) optimizes B-cell function via chaperoning integrin and TLR but not immunoglobulin. AB - Endoplasmic reticulum (ER) unfolded protein response (UPR) plays pivotal roles in both early B-cell development and plasma cell differentiation. As a major ER chaperone to mediate the UPR and a master chaperone for Toll-like receptors (TLRs), HSP90b1 (grp94, gp96) has long been implicated to facilitate the assembly of immunoglobulin. We hereby critically and comprehensively examine the roles of HSP90b1 in B-cell biology in vivo using B-cell-specific HSP90b1-null mice. We found that knockout B cells developed normally. There were no apparent problems with plasma cell differentiation, Ig assembly, class-switching, and Ig production. Strikingly, although both mutant conventional and innatelike B cells failed to compartmentalize properly due to loss of select but not all integrins, HSP90b1 was required for neither germinal center formation nor memory antibody responses in vivo. The only significant defect associated with HSP90b1 ablation in B cells was an attenuated antibody production in the context of TLR stimulation. Thus, our study has resolved the long-standing question regarding HSP90b1 in B-cell biology: HSP90b1 optimizes the function of B cells by chaperoning TLRs and integrins but not immunoglobulin. This study also has important implications in resolving the controversial roles of TLR in B-cell biology. PMID- 18509084 TI - Adoptive immunotherapy for indolent non-Hodgkin lymphoma and mantle cell lymphoma using genetically modified autologous CD20-specific T cells. AB - Adoptive immunotherapy with T cells expressing a tumor-specific chimeric T-cell receptor is a promising approach to cancer therapy that has not previously been explored for the treatment of lymphoma in human subjects. We report the results of a proof-of-concept clinical trial in which patients with relapsed or refractory indolent B-cell lymphoma or mantle cell lymphoma were treated with autologous T cells genetically modified by electroporation with a vector plasmid encoding a CD20-specific chimeric T-cell receptor and neomycin resistance gene. Transfected cells were immunophenotypically similar to CD8(+) effector cells and showed CD20-specific cytotoxicity in vitro. Seven patients received a total of 20 T-cell infusions, with minimal toxicities. Modified T cells persisted in vivo 1 to 3 weeks in the first 3 patients, who received T cells produced by limiting dilution methods, but persisted 5 to 9 weeks in the next 4 patients who received T cells produced in bulk cultures followed by 14 days of low-dose subcutaneous interleukin-2 (IL-2) injections. Of the 7 treated patients, 2 maintained a previous complete response, 1 achieved a partial response, and 4 had stable disease. These results show the safety, feasibility, and potential antitumor activity of adoptive T-cell therapy using this approach. This trial was registered at www.clinicaltrials.gov as #NCT00012207. PMID- 18509085 TI - Expression, activation, and function of integrin alphaMbeta2 (Mac-1) on neutrophil-derived microparticles. AB - Leukocyte-derived microparticles (MPs) are markers of cardiovascular diseases and contribute to pathogenesis by their interaction with various cell types. The presence and activation state of a multifunctional leukocyte receptor, integrin alpha(M)beta(2) (CD11b/18), on MPs derived from human neutrophils (PMNs) were examined. alpha(M)beta(2) expression was significantly enhanced on MPs derived from stimulated compared with resting PMNs. Furthermore, alpha(M)beta(2) on MPs from stimulated but not resting PMNs was in an activated conformation because it was capable of binding activation-specific monoclonal antibodies (CBRM1/5 and mAb24) and soluble fibrinogen. MPs expressing active alpha(M)beta(2) interacted with and were potent activators of resting platelets as assessed by induction of P-selectin expression and activation of alpha(IIb)beta(3). With the use of function-blocking antibodies and MPs obtained from alpha(M)(-/-)-deficient mice, we found that engagement of GPIbalpha on platelets by alpha(M)beta(2) on MPs plays a pivotal role in MP binding. Platelet activation by MPs occurs by a pathway dependent on Akt phosphorylation. PSGL-1/P-selectin interaction also is involved in the conjugation of MPs to platelets, and the combination of blocking reagents to both alpha(M)beta(2)/GPIbalpha and to PSGL-1/P-selectin completely abrogates MP-induced platelet activation. Thus, cooperation of these 2 receptor/counterreceptor systems regulates the prothrombotic properties of PMN derived MPs. PMID- 18509086 TI - Up-regulation of c-FLIP short by NFAT contributes to apoptosis resistance of short-term activated T cells. AB - Upon encounter with pathogens, T cells activate several defense mechanisms, one of which is the up-regulation of CD95 ligand (CD95L/FasL) which induces apoptosis in sensitive target cells. Despite expression of the CD95 receptor, however, recently activated T cells are resistant to CD95L, presumably due to an increased expression of antiapoptotic molecules. We show here that, in contrast to naive or long-term activated T cells, short-term activated T cells strongly up-regulate the caspase-8 inhibitor, cellular FLICE-inhibitory protein (c-FLIP). Intriguingly, upon activation, T cells highly induced the short splice variant c FLIP(short), whereas expression of c-FLIP(long) was only marginally modulated. In contrast to the general view that c-FLIP transcription is controlled predominantly by nuclear factor-kappaB (NF-kappaB), induction of c-FLIP(short) in T cells was primarily mediated by the calcineurin-nuclear factor of activated T cells (NFAT) pathway. Importantly, blockage of NFAT-mediated c-FLIP expression by RNA interference or inhibition of calcineurin rendered T cells sensitive toward CD95L, as well as activation-induced apoptosis. Thus, the resistance of recently activated T cells depends crucially on induction of c-FLIP expression by the calcineurin/NFAT pathway. Our findings imply that preventing autocrine CD95L signaling by c-FLIP facilitates T-cell effector function and an efficient immune response. PMID- 18509088 TI - Molecular profiling of pediatric mature B-cell lymphoma treated in population based prospective clinical trials. AB - The spectrum of entities, the therapeutic strategy, and the outcome of mature aggressive B-cell non-Hodgkin lymphomas (maB-NHLs) differs between children and adolescents on the one hand and adult patients on the other. Whereas adult maB NHLs have been studied in detail, data on molecular profiling of pediatric maB NHLs are hitherto lacking. We analyzed 65 cases of maB-NHL from patients up to 18 years of age by gene expression profiling, matrix comparative genomic hybridization (CGH), fluorescent in situ hybridization (FISH), and immunohistochemistry. The majority of the analyzed pediatric patients were treated within prospective trials (n = 49). We compared this group to a series of 182 previously published cases of adult maB-NHL. Gene expression profiling reclassified 31% of morphologically defined diffuse large B-cell lymphomas as molecular Burkitt lymphoma (mBL). The subgroups obtained by molecular reclassification did not show any difference in outcome in children treated with the NHL-Berlin-Frankfurt-Muenster (BFM) protocols. No differences were detectable between pediatric and adult mBL with regard to gene expression or chromosomal imbalances. This is the first report on molecular profiling of pediatric B-NHL showing mBL to be much more prominent in children than suggested by morphologic assessment. Based on molecular profiling mBL is a molecularly homogeneous disease across children and adults. PMID- 18509087 TI - Retrovirally induced CTL degranulation mediated by IL-15 expression and infection of mononuclear phagocytes in patients with HTLV-I-associated neurologic disease. AB - CD8(+) T cells contribute to central nervous system inflammation in human T-cell lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP). We analyzed CD8(+) T-cell dysfunction (degranulation and IFN-gamma production) and have demonstrated that CD8(+) T cells of patients with HAM/TSP (HAM/TSP patients) spontaneously degranulate and express IFN-gamma in ex vivo unstimulated culture. CD8(+) T cells of HTLV-I asymptomatic carriers and healthy donors did not. Spontaneous degranulation was detected in Tax11-19/HLA A*201 tetramer(+) cells, but not in CMV pp65 tetramer(+) cells. Interestingly, degranulation and IFN-gamma production in CD8(+) T cells was induced by coculture with autologous CD14(+) cells, but not CD4(+) T cells, of HAM/TSP patients, which correlated with proviral DNA load in CD14(+) cells of infected patients. Moreover, the expression of IL-15, which induced degranulation and IFN-gamma production in infected patients, was enhanced on surface of CD14(+) cells in HAM/TSP patients. Blockade of MHC class I and IL-15 confirmed these results. Thus, CD8(+) T-cell dysregulation was mediated by both virus infection and enhanced IL-15 on CD14(+) cells in HAM/TSP patients. Despite lower viral expression than in CD4(+) T cells, HTLV-I-infected or -activated CD14(+) cells may be a heretofore important but under recognized reservoir particularly in HAM/TSP patients. PMID- 18509089 TI - Immune system derived from homeostatic proliferation generates normal CD8 T-cell memory but altered repertoires and diminished heterologous immune responses. AB - The host responds to lymphopenic environments by acute homeostatic proliferation of T lymphocytes, which acquire phenotypes similar to memory cells. Using T-cell knockout (KO) mice adoptively reconstituted with splenocytes from immunologically naive mice, we examined the immune responses of an immune system derived from homeostatically proliferating (HP) T cells. HP cells mounted relatively normal acute CD8 T-cell responses to lymphocytic choriomeningitis virus (LCMV), but with altered T-cell receptor (TCR) repertoires, and they became functional memory cells capable of recall responses. Although homeostatic proliferation does not normally fully restore T-cell numbers, the CD8(+) T-cell pool was completely restored in T-cell KO mice after LCMV infection. CD4 T-cell responses were lower and not fully restored but seemed sufficient to allow for complete differentiation of CD8 memory T cells. The LCMV-immune HP mouse had an immune repertoire heavily biased with LCMV epitope-specific T cells with oligoclonal expansions. LCMV-immune HP mice had reduced cross-reactive and non-cross-reactive CD8 T-cell responses when challenged with a T cell-cross-reactive virus. Thus, whereas an HP immune system is capable of mounting relatively normal acute and memory CD8 T-cell responses, the narrowing of the T-cell repertoire may reduce immune responses to subsequently encountered pathogens. PMID- 18509090 TI - Heme oxygenase-1 deficiency leads to disrupted response to acute stress in stem cells and progenitors. AB - An effective response to extreme hematopoietic stress requires an extreme elevation in hematopoiesis and preservation of hematopoietic stem cells (HSCs). These diametrically opposed processes are likely to be regulated by genes that mediate cellular adaptation to physiologic stress. Herein, we show that heme oxygenase-1 (HO-1), the inducible isozyme of heme degradation, is a key regulator of these processes. Mice lacking one allele of HO-1 (HO-1(+/-)) showed accelerated hematopoietic recovery from myelotoxic injury, and HO-1(+/-) HSCs repopulated lethally irradiated recipients with more rapid kinetics. However, HO 1(+/-) HSCs were ineffective in radioprotection and serial repopulation of myeloablated recipients. Perturbations in key stem cell regulators were observed in HO-1(+/-) HSCs and hematopoietic progenitors (HPCs), which may explain the disrupted response of HO-1(+/-) HPCs and HPCs to acute stress. Control of stem cell stress response by HO-1 presents opportunities for metabolic manipulation of stem cell-based therapies. PMID- 18509091 TI - Epstein-Barr virus persistence in the absence of conventional memory B cells: IgM+IgD+CD27+ B cells harbor the virus in X-linked lymphoproliferative disease patients. AB - Epstein-Barr virus (EBV) persists in healthy virus carriers within the immunoglobulin (Ig)D(-)CD27(+) (class-switched) memory B-cell compartment that normally arises through antigen stimulation and germinal center transit. Patients with X-linked lymphoproliferative disease (XLP) lack such class-switched memory B cells but are highly susceptible to EBV infection, often developing fatal symptoms resembling those seen in EBV-associated hemophagocytic syndrome (EBV AHS), a disease caused by aberrant virus entry into the NK- or T-cell system. Here we show that XLP patients who survive primary EBV exposure carry relatively high virus loads in the B-cell, but not the NK- or T-cell, compartment. Interestingly, in the absence of conventional class-switched memory B cells, the circulating EBV load was concentrated within a small population of IgM(+)IgD(+)CD27(+) (nonswitched) memory cells rather than within the numerically dominant naive (IgM(+)IgD(+)CD27(-)) or transitional (CD10(+)CD27(-)) subsets. In 2 prospectively studied patients, the circulating EBV load was stable and markers of virus polymorphism detected the same resident strain over time. These results provide the first definitive evidence that EBV can establish persistence in the B cell system in the absence of fully functional germinal center activity and of a class-switched memory B-cell compartment. PMID- 18509092 TI - CNS aquaporin-4 autoimmunity in children. AB - BACKGROUND: In adult patients, autoantibodies targeting the water channel aquaporin-4 (AQP4) are a biomarker for a spectrum of CNS inflammatory demyelinating disorders with predilection for optic nerves and spinal cord (neuromyelitis optica [NMO]). Here we describe the neurologic, serologic, and radiographic findings associated with CNS AQP4 autoimmunity in childhood. METHODS: A total of 88 consecutive seropositive children were identified through service evaluation for NMO-IgG. Sera of 75 were tested for coexisting autoantibodies. Clinical information was available for 58. RESULTS: Forty-two patients (73%) were non-Caucasian, and 20 (34%) had African ethnicity. Median age at symptom onset was 12 years (range 4-18). Fifty-seven (98%) had attacks of either optic neuritis (n = 48; 83%) or transverse myelitis (n = 45; 78%), or both. Twenty-six (45%) had episodic cerebral symptoms (encephalopathy, ophthalmoparesis, ataxia, seizures, intractable vomiting, or hiccups). Thirty eight (68%) had brain MRI abnormalities, predominantly involving periventricular areas (in descending order of frequency): the medulla, supratentorial and infratentorial white matter, midbrain, cerebellum, thalamus, and hypothalamus. Additional autoantibodies were detected in 57 of 75 patients (76%), and 16 of 38 (42%) had a coexisting autoimmune disorder recorded (systemic lupus erythematosus, Sjogren syndrome, juvenile rheumatoid arthritis, Graves disease). Attacks were recurrent in 54 patients (93%; median follow-up, 12 months). Forty three of 48 patients (90%) had residual disability: 26 (54%) visual impairment and 21 (44%) motor deficits (median Expanded Disability Status Scale 4.0 at 12 months). CONCLUSIONS: Aquaporin-4 autoimmunity is a distinctive recurrent and widespread inflammatory CNS disease in children. PMID- 18509093 TI - No advantage of A beta 42-lowering NSAIDs for prevention of Alzheimer dementia in six pooled cohort studies. AB - INTRODUCTION: Observational studies show reduced incidence of Alzheimer dementia (AD) in users of nonsteroidal anti-inflammatory drugs (NSAIDs). One hypothesis holds that the subset of NSAIDs known as selective A beta(42)-lowering agents (SALAs) is responsible for this apparent reduction in AD risk. METHODS: We pooled individual-level data from six prospective studies to obtain a sufficient sample to examine AD risk in users of SALA vs non-SALA NSAIDs. RESULTS: Of 13,499 initially dementia-free participants (70,863 person-years), 820 developed incident AD. Users of NSAIDs (29.6%) showed reduced risk of AD (adjusted hazard ratio [aHR] 0.77, 95% CI 0.65-0.91). The point estimates were similar for SALAs (aHR 0.87, CI 0.72-1.04) and non-SALAs (aHR 0.75, CI 0.56-1.01). Because 573 NSAID users (14.5%) reported taking both a SALA and non-SALA, we examined their use alone and in combination. Resulting aHRs were 0.82 (CI 0.67-0.99) for SALA only, 0.60 (CI 0.40-0.90) for non-SALA only, and 0.87 (CI 0.57-1.33) for both NSAIDs (Wald test for differences, p = 0.32). The 40.7% of participants who used aspirin also showed reduced risk of AD, even when they used no other NSAIDs (aHR 0.78, CI 0.66-0.92). By contrast, there was no association with use of acetaminophen (aHR 0.93, CI 0.76-1.13). CONCLUSIONS: In this pooled dataset, nonsteroidal anti-inflammatory drug (NSAID) use reduced the risk of Alzheimer dementia (AD). However, there was no apparent advantage in AD risk reduction for the subset of NSAIDs shown to selectively lower A beta(42), suggesting that all conventional NSAIDs including aspirin have a similar protective effect in humans. PMID- 18509094 TI - Haplotypes and gene expression implicate the MAPT region for Parkinson disease: the GenePD Study. AB - BACKGROUND: Microtubule-associated protein tau (MAPT) has been associated with several neurodegenerative disorders including forms of parkinsonism and Parkinson disease (PD). We evaluated the association of the MAPT region with PD in a large cohort of familial PD cases recruited by the GenePD Study. In addition, postmortem brain samples from patients with PD and neurologically normal controls were used to evaluate whether the expression of the 3-repeat and 4-repeat isoforms of MAPT, and neighboring genes Saitohin (STH) and KIAA1267, are altered in PD cerebellum. METHODS: Twenty-one single-nucleotide polymorphisms (SNPs) in the region of MAPT on chromosome 17q21 were genotyped in the GenePD Study. Single SNPs and haplotypes, including the H1 haplotype, were evaluated for association to PD. Relative quantification of gene expression was performed using real-time RT-PCR. RESULTS: After adjusting for multiple comparisons, SNP rs1800547 was significantly associated with PD affection. While the H1 haplotype was associated with a significantly increased risk for PD, a novel H1 subhaplotype was identified that predicted a greater increased risk for PD. The expression of 4 repeat MAPT, STH, and KIAA1267 was significantly increased in PD brains relative to controls. No difference in expression was observed for 3-repeat MAPT. CONCLUSIONS: This study supports a role for MAPT in the pathogenesis of familial and idiopathic Parkinson disease (PD). Interestingly, the results of the gene expression studies suggest that other genes in the vicinity of MAPT, specifically STH and KIAA1267, may also have a role in PD and suggest complex effects for the genes in this region on PD risk. PMID- 18509095 TI - Biochemical markers in persons with preclinical familial Alzheimer disease. AB - BACKGROUND: Persons at risk for familial Alzheimer disease (FAD) provide a model in which biomarkers can be studied in presymptomatic disease. METHODS: Twenty-one subjects at risk for presenilin-1 (n = 17) or amyloid precursor protein (n = 4) mutations underwent evaluation with the Clinical Dementia Rating (CDR) scale. We obtained plasma from all subjects and CSF from 11. Plasma (Abeta(40), Abeta(42), F(2)-isoprostanes) and CSF (F(2)-isoprostanes, t-tau, p-tau(181), Abeta(40), Abeta(42), and Abeta(42)/Abeta(40) ratio) levels were compared between FAD mutation carriers (MCs) and noncarriers (NCs). RESULTS: Plasma Abeta(42) levels (25.1 pM vs 15.5 pM, p = 0.031) and the ratio of Abeta(42)/Abeta(40) (0.16 vs 0.11, p = 0.045) were higher in presymptomatic MCs. Among MCs, those with CDR scores of 0.5 had lower plasma Abeta(42) levels than those with CDR scores of 0 (14.1 pM vs 25.1, p = 0.02). The ratio of Abeta(42) to Abeta(40) was also reduced in the CSF (0.08 vs 0.15, p = 0.046) of nondemented MCs compared to NCs. Total CSF tau and p-tau(181) levels were elevated in presymptomatic FAD MCs. CSF levels of F(2)-isoprostanes were also elevated in MCs (n = 7, 48.6 pg/mL) compared to NCs (n = 4, 21.6 pg/mL, p = 0.031). CONCLUSIONS: Our data indicate that Abeta(42) is elevated in plasma in familial Alzheimer disease (FAD) mutation carriers (MCs) and suggests that this level may decrease with disease progression prior to the development of overt dementia. We also demonstrated that the ratio of Abeta(42) to Abeta(40) was reduced in the CSF of nondemented MCs and that elevations of t tau and p-tau(181) are sensitive indicators of presymptomatic disease. Our finding of elevated F(2)-isoprostane levels in the CSF of preclinical FAD MCs suggests that oxidative stress occurs downstream to mismetabolism of amyloid precursor protein. PMID- 18509096 TI - Highly lauded drug assessment program under attack. PMID- 18509097 TI - High-content assay to study protein prenylation. AB - The mevalonate pathway leads to synthesis of cholesterol and isoprenoid lipids. Prenyltransferases attach the isoprenoid lipids to the C-terminus of several small guanosine triphosphate-binding proteins. The prenyl groups are essential for the biological activity of these proteins. The prenyltransferases and other components of the mevalonate pathway are either present or potential drug targets for cancer, osteoporosis, restenosis, or high serum cholesterol level. Until recently, cellular assays to study protein prenylation have been tedious, low throughput assays. The authors have developed a high-content imaging-based assay to study protein prenylation. The assay is based on a green fluorescent protein (GFP) reporter, which is tagged with the prenylation motif of human H-Ras. The C terminus of H-Ras targets GFP to the plasma membrane. When protein prenylation is inhibited, the tagged GFP cannot be localized to plasma membrane but is soluble in the cells. The localization of the GFP reporter can be analyzed in the 96- or 384-well format using automated microscopy and automated image analysis. Information about cell number and nuclear intensity can be obtained from the same images. In compound screening, these readouts provide valuable information about the toxicity of the compounds. The authors have validated their assay using several inhibitors of the mevalonate pathway as well as siRNA against farnesyl pyrophosphate synthase, a critical enzyme in the synthesis of the isoprenoid lipids. PMID- 18509098 TI - Acid-sensitive channel inhibition prevents fetal alcohol spectrum disorders cerebellar Purkinje cell loss. AB - Ethanol is now considered the most common human teratogen. Educational campaigns have not reduced the incidence of ethanol-mediated teratogenesis, leading to a growing interest in the development of therapeutic prevention or mitigation strategies. On the basis of the observation that maternal ethanol consumption reduces maternal and fetal pH, we hypothesized that a pH-sensitive pathway involving the TWIK-related acid-sensitive potassium channels (TASKs) is implicated in ethanol-induced injury to the fetal cerebellum, one of the most sensitive targets of prenatal ethanol exposure. Pregnant ewes were intravenously infused with ethanol (258+/-10 mg/dl peak blood ethanol concentration) or saline in a "3 days/wk binge" pattern throughout the third trimester. Quantitative stereological analysis demonstrated that ethanol resulted in a 45% reduction in the total number of fetal cerebellar Purkinje cells, the cell type most sensitive to developmental ethanol exposure. Extracellular pH manipulation to create the same degree and pattern of pH fall caused by ethanol (manipulations large enough to inhibit TASK 1 channels), resulted in a 24% decrease in Purkinje cell number. We determined immunohistochemically that TASK 1 channels are expressed in Purkinje cells and that the TASK 3 isoform is expressed in granule cells of the ovine fetal cerebellum. Pharmacological blockade of both TASK 1 and TASK 3 channels simultaneous with ethanol effectively prevented any reduction in fetal cerebellar Purkinje cell number. These results demonstrate for the first time functional significance of fetal cerebellar two-pore domain pH-sensitive channels and establishes them as a potential therapeutic target for prevention of ethanol teratogenesis. PMID- 18509099 TI - Postconditioning for salvage of ischemic skeletal muscle from reperfusion injury: efficacy and mechanism. AB - We tested our hypothesis that postischemic conditioning (PostC) is effective in salvage of ischemic skeletal muscle from reperfusion injury and the mechanism involves inhibition of opening of the mitochondrial permeability transition pore (mPTP). In bilateral 8x13 cm pig latissimus dorsi muscle flaps subjected to 4 h ischemia, muscle infarction increased from 22+/-4 to 41+/-1% between 2 and 24 h reperfusion and remained unchanged at 48 (38+/-6%) and 72 (40+/-1%) h reperfusion (P<0.05; n=4 pigs). PostC induced by four cycles of 30-s reperfusion/reocclusion at the onset of reperfusion after 4 h ischemia reduced muscle infarction from 44+/-2 to 22+/-2% at 48 h reperfusion. This infarct protective effect of PostC was mimicked by intravenous injection of the mPTP opening inhibitor cyclosporin A or NIM-811 (10 mg/kg) at 5 min before the end of 4 h ischemia and was abolished by intravenous injection of the mPTP opener atractyloside (10 mg/kg) at 5 min before PostC (P<0.05; n=4-5 pigs). PostC or intravenous cyclosporin A injection at 5 min before reperfusion caused a decrease in muscle myeloperoxidase activity and mitochondrial free Ca2+ concentration and an increase in muscle ATP content after 4 h ischemia and 2 h reperfusion compared with the time-matched controls. These effects of PostC were abolished by intravenous injection of atractyloside at 5 min before PostC (P<0.05; n=6 pigs). These observations support our hypothesis that PostC is effective in salvage of ischemic skeletal muscle from reperfusion injury and the mechanism involves inhibition of opening of the mPTP. PMID- 18509100 TI - Ventilatory responses to acute and chronic hypoxia are altered in female but not male Paskin-deficient mice. AB - Proteins harboring a Per-Arnt-Sim (PAS) domain are versatile and allow archaea, bacteria, and plants to sense oxygen partial pressure, as well as light intensity and redox potential. A PAS domain associated with a histidine kinase domain is found in FixL, the oxygen sensor molecule of Rhizobium species. PASKIN is the mammalian homolog of FixL, but its function is far from being understood. Using whole body plethysmography, we evaluated the ventilatory response to acute and chronic hypoxia of homozygous deficient male and female PASKIN mice (Paskin-/-). Although only slight ventilatory differences were found in males, female Paskin-/ mice increased ventilatory response to acute hypoxia. Unexpectedly, females had an impaired ability to reach ventilatory acclimatization in response to chronic hypoxia. Central control of ventilation occurs in the brain stem respiratory centers and is modulated by catecholamines via tyrosine hydroxylase (TH) activity. We observed that TH activity was altered in male and female Paskin-/- mice. Peripheral chemoreceptor effects on ventilation were evaluated by exposing animals to hyperoxia (Dejours test) and domperidone, a peripheral ventilatory stimulant drug directly affecting the carotid sinus nerve discharge. Male and female Paskin-/- had normal peripheral chemosensory (carotid bodies) responses. In summary, our observations suggest that PASKIN is involved in the central control of hypoxic ventilation, modulating ventilation in a gender-dependent manner. PMID- 18509101 TI - Early fetal hypoxia leads to growth restriction and myocardial thinning. AB - Hypoxia is necessary for fetal development; however, excess hypoxia is detrimental. Hypoxia has been extensively studied in the near-term fetus, but less is known about earlier fetal effects. The purpose of this study was to determine the window of vulnerability to severe hypoxia, what organ system(s) is most sensitive, and why hypoxic fetuses die. We induced hypoxia by reducing maternal-inspired O2 from 21% to 8%, which decreased fetal tissue oxygenation assessed by pimonidazole binding. The mouse fetus was most vulnerable in midgestation: 24 h of hypoxia killed 89% of embryonic day 13.5 (E13.5) fetuses, but only 5% of E11.5 and 51% of E17.5 fetuses. Sublethal hypoxia at E12.5 caused growth restriction, reducing fetal weight by 26% and protein by 45%. Hypoxia induced HIF-1 target genes, including vascular endothelial growth factor (Vegf), erythropoietin, glucose transporter-1 and insulin-like growth factor binding protein-1 (Igfbp-1), which has been implicated in human intrauterine growth restriction (IUGR). Hypoxia severely compromised the cardiovascular system. Signs of heart failure, including loss of yolk sac circulation, hemorrhage, and edema, were caused by 18-24 h of hypoxia. Hypoxia induced ventricular dilation and myocardial hypoplasia, decreasing ventricular tissue by 50% and proliferation by 21% in vivo and by 40% in isolated cultured hearts. Epicardial detachment was the first sign of hypoxic damage in the heart, although expression of epicardially derived mitogens, such as FGF2, FGF9, and Wnt9b was not reduced. We propose that hypoxia compromises the fetus through myocardial hypoplasia and reduced heart rate. PMID- 18509102 TI - Influence of sedentary versus physically active conditions on regulation of plasma renin activity and vasopressin. AB - Physical inactivity is an independent risk factor for cardiovascular disease. Sedentary animals compared to physically active controls exhibit enhanced sympathoexcitatory responses, including arterial baroreflex-mediated sympathoexcitation. Hypotension-induced sympathoexcitation is also associated with the release of vasoactive hormones. We hypothesized that sedentary conditions may enhance release of the vasoactive hormones AVP and ANG II. To test this hypothesis, the humoral response to hypotension was examined in conscious rats after 9-12 wk of sedentary conditions or "normally active" conditions. Normally active conditions were produced by allowing rats access to running wheels in their home cages. Running distance peaked after 4 wk (4.5 +/- 0.7 km/day), and the total distance run after 9 wk was 174 +/- 23 km (n = 25). Similar levels of hypotension were induced in conscious sedentary or physically active animals with the arterial vasodilator, diazoxide (25 mg/kg iv). Control experiments used a saline injection of equivalent volume. Plasma samples were collected and assayed for plasma AVP concentration and plasma renin activity (PRA). Sedentary conditions significantly enhanced resting and hypotension induced PRA relative to normal physical activity. In contrast, resting and hypotension-induced AVP levels were not statistically different between groups. These data suggest that baroreflex-mediated activation of the renin-angiotensin system, but not AVP secretion, is enhanced by sedentary conditions. We speculate that augmented activation of the renin-angiotensin system may be related to enhanced sympathetic outflow observed in sedentary animals and may contribute to increased risk of cardiovascular disease in the sedentary population. PMID- 18509103 TI - Sex-specific differences in chromosome-dependent regulation of vascular reactivity in female consomic rat strains from a SSxBN cross. AB - High-throughput studies in the Medical College of Wisconsin Program for Genomic Applications (Physgen) were designed to link chromosomes with physiological function in consomic strains derived from a cross between Dahl salt-sensitive SS/JrHsdMcwi (SS) and Brown Norway normotensive BN/NHsdMcwi (BN) rats. The specific goal of the vascular protocol was to characterize the responses of aortic rings from these strains to vasoconstrictor and vasodilator stimuli (phenylephrine, acetylcholine, sodium nitroprusside, and bath hypoxia) to identify chromosomes that either increase or decrease vascular reactivity to these vasoactive stimuli. Because previous studies demonstrated sex-specific quantitative trait loci (QTLs) related to regulation of cardiovascular phenotypes in an F2 cross between the parental strains, males and females of each consomic strain were included in all experiments. As there were significant sex-specific differences in aortic sensitivity to vasoconstrictor and vasodilator stimuli compared with the parental SS strain, we report the results of the females separately from the males. There were also sex-specific differences in aortic ring sensitivity to these vasoactive stimuli in consomic strains that were fed a high-salt diet (4% NaCl) for 3 wk to evaluate salt-induced changes in vascular reactivity. Differences in genetic architecture could contribute to sex-specific differences in the development and expression of cardiovascular diseases via differential regulation and expression of genes. Our findings are the first to link physiological traits with specific chromosomes in female SS rats and support the idea that sex is an important environmental variable that plays a role in the expression and regulation of genes. PMID- 18509104 TI - Antibodies against GM1 in demented patients. AB - The aim of this study was to evaluate the levels of anti-GM1 in demented patients, correlating them with the type and severity of dementia as well as with the eventually coexistent polyneuropathy. Anti-GM1 concentrations were measured in the sera of 33 demented patients with a male-to-female ratio of 1:2.7 (the mean age was 69.7 years for males and 70.1 years for females). Eighty-two percent of the patients revealed increased values of anti-GM1, but only 18.2% demonstrated polyneuropathies. Fifty-nine percent of the patients suffered from vascular dementia. The most severely demented patients demonstrated a Mini-Mental State Examination score of 5 to 23 out of 30 and revealed the most increased levels of anti-GM1 (>40 EU/mL). The findings of this study are indicative of a possible correlation between the levels of anti-GM1 and the severity of dementia, mainly of the vascular type. PMID- 18509105 TI - Consensus statement on genetic research in dementia. AB - In this article, the authors describe how the European Dementia Consensus Network developed a consensus on research ethics in dementia, taking into account the questions posed by the era of genetic research and its new research methods. The consensus process started with a Delphi procedure to analyze relevant stakeholders' positions by describing their statements on the possibilities and limitations of research into genetic determinants of Alzheimer disease and to describe and analyze the moral desirability of genetic research on Alzheimer disease. The conclusions drawn from the Delphi procedure fuelled the development of the consensus statement, which is presented in this paper. The consensus statement aims to stimulate ethically acceptable research in the field of dementia and the protection of vulnerable elderly patients with dementia from application of inadequate research methods or designs. PMID- 18509106 TI - Pharmacotherapy for inappropriate sexual behaviors in dementia: a systematic review of literature. AB - The aim of this study is to systematically review the published literature on pharmacotherapy for inappropriate sexual behaviors in dementia. Literature search of the 5 databases (PubMed, MEDLINE, EMBASE, PsychINFO, and COCHRANE collaboration) and the analysis of the data available for the pharmacotherapeutic treatments of inappropriate sexual behaviors in dementia were carried out. There are no published randomized controlled trials of pharmacotherapy for inappropriate sexual behaviors in dementia, but available data form uncontrolled trials, case series, and individual case reports suggest efficacy for antidepressants, antipsychotics, mood stabilizers, hormonal agents, cimetidine, and pindolol for the treatment of these behaviors. Although there are no controlled data for the treatment of inappropriate sexual behaviors in dementia, available data suggest efficacy for some commonly used pharmacotherapeutic agents. PMID- 18509107 TI - Functional properties of the carboxy-terminal host cell-binding domains of the two toxins, TcdA and TcdB, expressed by Clostridium difficile. AB - The biological and ligand-binding properties of recombinant C-terminal cell binding domains (CBDs) and subdomains of the two large exotoxins, Toxin A (TcdA) and Toxin B (TcdB) expressed by Clostridium difficile were examined in the hemagglutination and Verocytotoxicity neutralization assays and by qualitative affinity chromatography using Sepharose-linked alpha Gal(1,3)betaGal(1,4)beta Glc as well as the direct electrospray ionization mass spectrometry (ES-MS) assay. These studies revealed that, whereas the full-length TcdA CBD agglutinated rabbit erythrocytes, neutralized TcdA-mediated Vero cell death and bound to alpha Gal(1,3)betaGal(1,4)beta Glc-derivatized Sepharose, the TcdB CBD was inactive in these functional assays. Moreover, retention by alpha Gal(1,3)betaGal(1,4)beta Glc-derivatized Sepharose corresponded to the number of available TcdA subdomain ligand-binding sites. By contrast, the ES-MS assays revealed that both the TcdA and TcdB CBD bind to 8-methoxycarbonyloctyl-alpha Gal(1,3)betaGal(1,4)beta Glc sequences with similar avidities. Additional ES-MS experiments using chemically altered alpha Gal(1,3)betaGal(1,4)beta Glc sequences also revealed that the TcdA and TcdB CBD will tolerate a fair amount of structural variation in their complementary glycan ligands. Although the studies are consistent with the known ligand-binding properties of the TcdA and TcdB holotoxins, they also revealed subtle heretofore unrecognized functional differences in their receptor recognition properties. PMID- 18509108 TI - Multifunctionality of Campylobacter jejuni sialyltransferase CstII: characterization of GD3/GT3 oligosaccharide synthase, GD3 oligosaccharide sialidase, and trans-sialidase activities. AB - CstII from bacterium Campylobacter jejuni strain OH4384 has been previously characterized as a bifunctional sialyltransferase having both alpha2,3 sialyltransferase (GM3 oligosaccharide synthase) and alpha2,8-sialyltransferase (GD3 oligosaccharide synthase) activities which catalyze the transfer of N acetylneuraminic acid (Neu5Ac) from cytidine 5'-monophosphate (CMP)-Neu5Ac to C 3' of the galactose in lactose and to C-8 of the Neu5Ac in 3'-sialyllactose, respectively (Gilbert M, Karwaski MF, Bernatchez S, Young NM, Taboada E, Michniewicz J, Cunningham AM, Wakarchuk WW. 2002. The genetic bases for the variation in the lipo-oligosaccharide of the mucosal pathogen, Campylobacter jejuni. Biosynthesis of sialylated ganglioside mimics in the core oligosaccharide. J Biol Chem. 277:327-337). We report here the characterization of a truncated CstII mutant (CstIIDelta32(I53S)) cloned from a synthetic gene whose codons are optimized for an Escherichia coli expression system. In addition to the alpha2,3- and alpha2,8-sialyltransferase activities reported before for the synthesis of GM3- and GD3-type oligosaccharides, respectively, the CstIIDelta32(I53S) has alpha2,8-sialyltransferase (GT3 oligosaccharide synthase) activity for the synthesis of GT3 oligosaccharide. It also has alpha2,8-sialidase (GD3 oligosaccharide sialidase) activity that catalyzes the specific cleavage of the alpha2,8-sialyl linkage of GD3-type oligosaccharides and alpha2,8-trans sialidase (GD3 oligosaccharide trans-sialidase) activity that catalyzes the transfer of a sialic acid from a GD3 oligosaccharide to a different GM3 oligosaccharide (3'-sialyllactoside). The donor substrate specificity study of the CstIIDelta32(I53S) GD3 oligosaccharide synthase activity indicates that the enzyme is flexible in using different CMP-activated sialic acids and their analogs for the synthesis of GD3 oligosaccharides containing natural and nonnatural modifications at the terminal sialic acid. PMID- 18509109 TI - Human and mouse macrophage-inducible C-type lectin (Mincle) bind Candida albicans. AB - Candida albicans is a causative agent in mycoses of the skin, oral cavity, and gastrointestinal tract. Identification of receptors, and their respective ligands, that are engaged by immune cells when in contact with C. albicans is crucial for understanding inflammatory responses leading to invasive candidiasis. Mincle is a recently identified macrophage-expressed receptor that is important for host responses to C. albicans. The carbohydrate-recognition domain of human and mouse Mincle were expressed, purified under denaturing conditions, and successfully refolded. In addition to oligomers, there are isolatable monomeric and dimeric forms of the protein that occur under two different buffer solutions. The human and mouse homologues bound yeast extract, and the isolated dimeric and monomeric species also demonstrated the recognition of whole C. albicans yeast cells. The data are indicative of several functional states mediating the interaction of Mincle and yeast at the surface of the macrophage. PMID- 18509110 TI - Temperature-dependent cooperativity in donor-acceptor substrate binding to the human blood group glycosyltransferases. AB - Affinities of the human blood group glycosyltransferases, alpha-(1-->3)-N acetylgalactosaminyltransferase (GTA) and alpha-(1-->3)-galactosyltransferase (GTB) for their common acceptor substrate alpha-l-Fucp-(1-->2)-beta-d-Galp O(CH2)(7)CH3 (1), in the absence and presence of bound uridine 5'-diphosphate (UDP) and Mn2+ were determined using temperature-controlled electrospray ionization mass spectrometry. The presence of bound UDP and Mn(2+) in the donor binding site has a marked influence on the thermodynamic parameters for the association of 1 with GTA and GTB. Both the enthalpy and entropy of association (DeltaH(a), DeltaS(a)) decrease significantly. However, the free energy of association (DeltaG(a)) is unchanged at physiological temperature. The differences in the DeltaH(a) and DeltaS(a) values determined in the presence and absence of bound UDP are attributed to structural changes in the glycosyltransferases induced by the simultaneous binding of 1 and UDP. PMID- 18509111 TI - Contextual specificity of extinction of delay but not trace eyeblink conditioning in humans. AB - Renewal of an extinguished conditioned response has been demonstrated in humans and in animals using various types of procedures, except renewal of motor learning such as eyeblink conditioning. We tested renewal of delay and trace eyeblink conditioning in a virtual environment in an ABA design. Following acquisition in one context (A, e.g., an airport) and extinction in a different context (B, e.g., a city), tests for renewal took place in the acquisition (A) and extinction context (B), in a counterbalanced order. Results showed renewal of the extinguished conditioned response in the delay but not trace condition. PMID- 18509112 TI - Reactivation-dependent amnesia for appetitive memories is determined by the contingency of stimulus presentation. AB - Previously acquired aversive and appetitive memories are not stable and permanent. The reactivation of such memories by re-exposure to training stimuli renders them vulnerable to disruption by amnestic agents such as the noncompetitive N-methyl-D-aspartate receptor antagonist (+)-5-methyl-10,11 dihydro-SH-dibenzo{a,d}cyclohepten-5,10-imine maleate (MK-801). However, relatively little is known about the parameters that influence the reactivation process. Here, we show that the method of stimulus presentation during memory reactivation is of great importance. Male Lister Hooded rats were trained to acquire a lever press response that delivered a sucrose reward paired with a light conditioned stimulus (CS). The CS-sucrose association was then reactivated through re-exposure to the CS, either contingently upon the lever press response, or noncontingently in the absence of instrumental responding. Systemic administration of MK-801 (0.1 mg/kg) at the time of memory reactivation resulted in amnesia, and hence a reduction in subsequent sucrose seeking induced by, and dependent upon, presentation of the CS, only when the memory was reactivated contingently. Therefore, stimuli may have to be presented in the same manner at memory reactivation as during learning in order to render a previously acquired memory vulnerable to disruption. These results have important implications for the potential translational use of glutamatergic treatments in conjunction with targeted memory reactivation. PMID- 18509113 TI - Evidence for recovery of fear following immediate extinction in rats and humans. AB - Fear responses can be eliminated through extinction, a procedure involving the presentation of fear-eliciting stimuli without aversive outcomes. Extinction is believed to be mediated by new inhibitory learning that acts to suppress fear expression without erasing the original memory trace. This hypothesis is supported mainly by behavioral data demonstrating that fear can recover following extinction. However, a recent report by Myers and coworkers suggests that extinction conducted immediately after fear learning may erase or prevent the consolidation of the fear memory trace. Since extinction is a major component of nearly all behavioral therapies for human fear disorders, this finding supports the notion that therapeutic intervention beginning very soon after a traumatic event will be more efficacious. Given the importance of this issue, and the controversy regarding immediate versus delayed therapeutic interventions, we examined two fear recovery phenomena in both rats and humans: spontaneous recovery (SR) and reinstatement. We found evidence for SR and reinstatement in both rats and humans even when extinction was conducted immediately after fear learning. Thus, our data do not support the hypothesis that immediate extinction erases the original memory trace, nor do they suggest that a close temporal proximity of therapeutic intervention to the traumatic event might be advantageous. PMID- 18509114 TI - Activation of exchange protein activated by cyclic-AMP enhances long-lasting synaptic potentiation in the hippocampus. AB - cAMP is a critical second messenger implicated in synaptic plasticity and memory in the mammalian brain. Substantial evidence links increases in intracellular cAMP to activation of cAMP-dependent protein kinase (PKA) and subsequent phosphorylation of downstream effectors (transcription factors, receptors, protein kinases) necessary for long-term potentiation (LTP) of synaptic strength. However, cAMP may also initiate signaling via a guanine nucleotide exchange protein directly activated by cAMP (Epac). The role of Epac in hippocampal synaptic plasticity is unknown. We found that in area CA1 of mouse hippocampal slices, activation of Epac enhances maintenance of LTP without affecting basal synaptic transmission. The persistence of this form of LTP requires extracellular signal-regulated protein kinase (ERK) and new protein synthesis, but not transcription. Because ERK is involved in translational control of long-lasting plasticity and memory, our data suggest that Epac is a crucial link between cAMP and ERK during some forms of protein synthesis-dependent LTP. Activation of Epac represents a novel signaling pathway for rapid regulation of the stability of enduring forms of LTP and, perhaps, of hippocampus- dependent long-term memories. PMID- 18509116 TI - Videos in clinical medicine. Cricothyroidotomy. PMID- 18509115 TI - Training with inedible food in Aplysia causes expression of C/EBP in the buccal but not cerebral ganglion. AB - Training with inedible food in Aplysia increased expression of the transcription factor C/EBP in the buccal ganglia, which primarily have a motor function, but not in the cerebral or pleural ganglia. C/EBP mRNA increased immediately after training, as well as 1-2 h later. The increased expression of C/EBP protein lagged the increase in mRNA. Stimulating the lips and inducing feeding responses do not lead to long-term memory and did not cause increased C/EBP expression. Blocking polyADP-ribosylation, a process necessary for long-term memory after training, did not affect the increased C/EBP mRNA expression in the buccal ganglia. PMID- 18509117 TI - Images in clinical medicine. Severe constipation. PMID- 18509118 TI - The moral of the story. PMID- 18509119 TI - Exploiting a research underclass in phase 1 clinical trials. PMID- 18509120 TI - Methylnaltrexone for opioid-induced constipation in advanced illness. AB - BACKGROUND: Constipation is a distressing side effect of opioid treatment. As a quaternary amine, methylnaltrexone, a mu-opioid-receptor antagonist, has restricted ability to cross the blood-brain barrier. We investigated the safety and efficacy of subcutaneous methylnaltrexone for treating opioid-induced constipation in patients with advanced illness. METHODS: A total of 133 patients who had received opioids for 2 or more weeks and who had received stable doses of opioids and laxatives for 3 or more days without relief of opioid-induced constipation were randomly assigned to receive subcutaneous methylnaltrexone (at a dose of 0.15 mg per kilogram of body weight) or placebo every other day for 2 weeks. Coprimary outcomes were laxation (defecation) within 4 hours after the first dose of the study drug and laxation within 4 hours after two or more of the first four doses. Patients who completed this phase were eligible to enter a 3 month, open-label extension trial. RESULTS: In the methylnaltrexone group, 48% of patients had laxation within 4 hours after the first study dose, as compared with 15% in the placebo group, and 52% had laxation without the use of a rescue laxative within 4 hours after two or more of the first four doses, as compared with 8% in the placebo group (P<0.001 for both comparisons). The response rate remained consistent throughout the extension trial. The median time to laxation was significantly shorter in the methylnaltrexone group than in the placebo group. Evidence of withdrawal mediated by central nervous system opioid receptors or changes in pain scores was not observed. Abdominal pain and flatulence were the most common adverse events. CONCLUSIONS: Subcutaneous methylnaltrexone rapidly induced laxation in patients with advanced illness and opioid-induced constipation. Treatment did not appear to affect central analgesia or precipitate opioid withdrawal. (Clinical Trials.gov number, NCT00402038 [ClinicalTrials.gov].). PMID- 18509121 TI - A placebo-controlled trial of prucalopride for severe chronic constipation. AB - BACKGROUND: In this 12-week trial, we aimed to determine the efficacy of prucalopride, a selective, high-affinity 5-hydroxytryptamine4 receptor agonist, in patients with severe chronic constipation. METHODS: In our multicenter, randomized, placebo-controlled, parallel-group, phase 3 trial, patients with severe chronic constipation (< or =2 spontaneous, complete bowel movements per week) received placebo or 2 or 4 mg of prucalopride, once daily, for 12 weeks. The primary efficacy end point was the proportion of patients having three or more spontaneous, complete bowel movements per week, averaged over 12 weeks. Secondary efficacy end points were derived from daily diaries and validated questionnaires completed by patients. Adverse events, clinical laboratory values, and cardiovascular effects were monitored. RESULTS: Efficacy was analyzed in 620 patients. The proportion of patients with three or more spontaneous, complete bowel movements per week was 30.9% of those receiving 2 mg of prucalopride and 28.4% of those receiving 4 mg of prucalopride, as compared with 12.0% in the placebo group (P<0.001 for both comparisons). Over 12 weeks, 47.3% of patients receiving 2 mg of prucalopride and 46.6% of those receiving 4 mg of prucalopride had an increase in the number of spontaneous, complete bowel movements of one or more per week, on average, as compared with 25.8% in the placebo group (P<0.001 for both comparisons). All other secondary efficacy end points, including patients' satisfaction with their bowel function and treatment and their perception of the severity of their constipation symptoms, were significantly improved with the use of 2 or 4 mg of prucalopride as compared with placebo, at week 12. The most frequent treatment-related adverse events were headache and abdominal pain. There were no significant cardiovascular effects of treatment. CONCLUSIONS: Over 12 weeks, prucalopride significantly improved bowel function and reduced the severity of symptoms in patients with severe chronic constipation. Larger and longer trials are required to further assess the risks and benefits of the use of prucalopride for chronic constipation. (ClinicalTrials.gov number, NCT00483886 [ClinicalTrials.gov].). PMID- 18509122 TI - Clinical practice. Precocious puberty. PMID- 18509123 TI - Hepatopulmonary syndrome--a liver-induced lung vascular disorder. PMID- 18509124 TI - Images in clinical medicine. The aorta in osteoporosis. PMID- 18509125 TI - Case records of the Massachusetts General Hospital. Case 17-2008. A 63-year-old man with metastatic renal-cell carcinoma. PMID- 18509126 TI - Opioid side effects--mechanism-based therapy. PMID- 18509127 TI - The long and short of a constipation-reducing medication. PMID- 18509128 TI - Local paclitaxel delivery in peripheral vascular disease. PMID- 18509129 TI - Cardiovascular events during World Cup soccer. PMID- 18509130 TI - Hutchinson-Gilford progeria syndrome. PMID- 18509131 TI - Effect of cost sharing on screening mammography. PMID- 18509132 TI - Systemic lupus erythematosus. PMID- 18509133 TI - Acute lower respiratory tract infection. PMID- 18509134 TI - Cortical arousal in children with severe enuresis. PMID- 18509135 TI - Body composition indices in women with well-controlled type 1 diabetes. PMID- 18509136 TI - Reversible focal hepatic steatosis in type 1 diabetic patients treated with intraperitoneal insulin implantable pump therapy. PMID- 18509137 TI - Diabetic ketoacidosis induced by aripiprazole in a 12-year-old boy. PMID- 18509138 TI - Influence of flickering light on the retinal vessels in diabetic patients: response to Mandecka et al. PMID- 18509140 TI - Abnormal liver function test predicts type 2 diabetes: a community-based prospective study: response to Cho et al. PMID- 18509141 TI - Diabetes, depression, and death: a randomized controlled trial of a depression treatment program for older adults based in primary care (PROSPECT): response to Bogner et al. PMID- 18509143 TI - A step backward--or is it forward? PMID- 18509144 TI - Cardiovascular disease in diabetes. PMID- 18509145 TI - HIV therapy and diabetes risk. PMID- 18509146 TI - Effect of 80-hour workweek on continuity of care. AB - Work limitations were mandated (2003) to increase safety and improve resident lifestyle. Is clinic continuity affected? Medical University of South Carolina pediatric residents' records for 6 months of 2002 (before regulation) and 2003 (after regulation) were reviewed. Continuity for physician formula, t tests, and multivariate linear regression were used. Continuity was calculated for 44 residents (2002) and 45 residents (2003). Mean continuity was 54% (2002) and 53% (2003; P = .5); continuity for well-child care visits was 78% (2002) and 73% (2003; P = .047). Continuity decreased most for interns (52% [2002], 47% [2003] for all visits; 76%, 67% for well-child care visits). In the multivariate model, year did not predict continuity. When only well-child care visits were considered, year showed a trend toward significance ( P = .07): 2003 had less continuity. Compared with third-year residents, interns had 8% points less continuity for all visits (6% points less for well-child care visits). Continuity can be maintained despite regulations. Interns are most vulnerable. PMID- 18509147 TI - Millipede burn at an unusual site mimicking child abuse in an 8-year-old girl. PMID- 18509148 TI - Use of the rapid antigenic test to determine the duration of isolation in infants hospitalized for respiratory syncytial virus infections. AB - Infants hospitalized for bronchiolitis associated with respiratory syncytial virus are isolated for the duration of their hospitalization. This article reviews whether a negative rapid antigenic test could be used to shorten the duration of isolation measures. Rapid respiratory syncytial virus tests were performed from day 3 on alternate days in all patients hospitalized for bronchiolitis. Isolation measures were removed when the test result was negative. The result of the antigenic test was confirmed by viral culture and polymerase chain reaction. Surveillance of nosocomial cases was performed daily. Forty-one patients were analyzed. On day 3, 51.2% of patients were negative by viral culture and 56.1% were negative by rapid testing. On day 5, a further 26.8% were negative by viral culture and 31.7% by rapid testing. The rapid antigen test had a low sensitivity at 60% and a specificity of 76% compared with viral cultures; therefore this test alone cannot be used to lift isolation measures. PMID- 18509149 TI - Protein-energy malnutrition and feeding refusal secondary to food allergies. PMID- 18509150 TI - Pediatric providers' acceptance of recommendations for routine rotavirus vaccination. PMID- 18509151 TI - Methicillin-resistant Staphylococcus aureus endocarditis in an extremely low birth-weight infant treated with linezolid. PMID- 18509152 TI - Haemophilus influenzae type B disease: have we forgotten? PMID- 18509153 TI - Lemierre's syndrome and defensive antibiotic prescribing tactics: lessons to be had. PMID- 18509154 TI - Chronic illness. PMID- 18509155 TI - Fever, rash, and hepatic dysfunction in a 3-year-old child: a case report. PMID- 18509156 TI - Ask Dr. Sue: little boys in men's locker rooms. PMID- 18509159 TI - Particulate urban air pollution affects the functional morphology of mouse placenta. AB - In humans, adverse pregnancy outcomes (low birth weight, prematurity, and intrauterine growth retardation) are associated with exposure to urban air pollution. Experimental data have also shown that such exposure elicits adverse reproductive outcomes. We hypothesized that the effects of urban air pollution on pregnancy outcomes could be related to changes in functional morphology of the placenta. To test this, future dams were exposed during pregestational and gestational periods to filtered or nonfiltered air in exposure chambers. Placentas were collected from near-term pregnancies and prepared for microscopical examination. Fields of view on vertical uniform random tissue slices were analyzed using stereological methods. Volumes of placental compartments were estimated, and the labyrinth was analyzed further in terms of its maternal vascular spaces, fetal capillaries, trophoblast, and exchange surface areas. From these primary data, secondary quantities were derived: vessel calibers (expressed as diameters), trophoblast thickness (arithmetic mean), and total and mass-specific morphometric diffusive conductances for oxygen of the intervascular barrier. Two-way analysis of variance showed that both periods of exposure led to significantly smaller fetal weights. Pregestational exposure to nonfiltered air led to significant increases in fetal capillary surface area and in total and mass-specific conductances. However, the calibers of maternal blood spaces were reduced. Gestational exposure to nonfiltered air was associated with reduced volumes, calibers, and surface areas of maternal blood spaces and with greater fetal capillary surfaces and diffusive conductances. The findings indicate that urban air pollution affects placental functional morphology. Fetal weights are compromised despite attempts to improve diffusive transport across the placenta. PMID- 18509160 TI - Nuclear transfer-mediated rescue of the nuclear genome of nonviable mouse cells frozen without cryoprotectant. AB - Nuclear transfer (NT) provides an opportunity for clonal amplification of a nuclear genome of interest. Here, we report NT-mediated reprogramming with frozen mouse cells that were nonviable because they were frozen at -80 degrees C for up to 342 days without a cryoprotectant. We derived eight embryonic stem (ES) cell lines from cloned blastocysts by conventional NT procedure and five ntES (nuclear transfer embryonic stem) cell lines by a modified NT procedure in which a whole cell instead of a nucleus was injected into an enucleated oocyte. Chromosome analysis revealed that 12 of 13 ntES cell lines have normal karyotypes. On injection of ntES cells into tetraploid blastocysts to generate clonal mice that are nearly completely ntES-cell derived, live pups were obtained; four clonal mice survived until adulthood. On injection of ntES cells into diploid blastocysts, chimeric mice with a high somatic ES cell contribution were generated; germ-line transmission was obtained. Our findings indicate that chromosome stability and genomic integrity can be maintained in mouse somatic cells after freezing without cryoprotection and that NT and ES cell techniques can rescue the genome of these cells. PMID- 18509161 TI - Lim homeobox gene, lhx8, is essential for mouse oocyte differentiation and survival. AB - Lhx8 is a member of the LIM-homeobox transcription factor family and preferentially expressed in oocytes and germ cells within the mouse ovary. We discovered that Lhx8 knockout females lose oocytes within 7 days after birth. At the time of birth, histological examination shows that Lhx8-deficient (Lhx8-/-) ovaries are grossly similar to the newborn wild-type ovaries. Lhx8-/- ovaries fail to maintain the primordial follicles, and the transition from primordial to growing follicles does not occur. Lhx8-/- ovaries misexpress oocyte-specific genes, such as Gdf9, Pou5f1, and Nobox. Very rapid loss of oocytes may partly be due to the drastic downregulation of Kit and Kitl in Lhx8-/- ovaries. We compared Lhx8-/- and wild-type ovaries using an Affymetrix 430 2.0 microarray platform. A total of 80 (44%) of 180 of the genes downregulated more than 5-fold in Lhx8-/- ovaries were preferentially expressed in oocytes, whereas only 3 (2%) of 146 genes upregulated more than 5-fold in the absence of Lhx8 were preferentially expressed in oocytes. In addition, the comparison of genes regulated in Lhx8-/- and Nobox-/- newborn ovaries discovered a common set of 34 genes whose expression level was affected in both Lhx8- and Nobox-deficient mice. Our findings show that Lhx8 is a critical factor for maintenance and differentiation of the oocyte during early oogenesis, and it acts in part by downregulating the Nobox pathway. PMID- 18509162 TI - Protein phosphatase 3 differentially modulates vascular endothelial growth factor and fibroblast growth factor 2-stimulated cell proliferation and signaling in ovine fetoplacental artery endothelial cells. AB - A critical process for vascular endothelial growth factor (VEGF)- and fibroblast growth factor 2 (FGF2)-regulated cellular function is reversible protein phosphorylation, which is tightly controlled by a balance of protein kinases and phosphatases. We have reported that in ovine fetoplacental artery endothelial (OFPAE) cells, VEGF and FGF2 stimulate cell proliferation in part via activation of mitogen-activated protein kinase kinase 1/2 (MAP2K1/2)/mitogen-activated protein kinase 3/1 (MAPK3/1) and phosphoinositide 3-kinase (PI3K)/v-akt murine thymoma viral oncogene homolog 1 (AKT1) pathways. In the present study, we examined if protein phosphatase 3 (PPP3) mediated VEGF- and FGF2-stimulated OFPAE cell proliferation via modulating activation of MAPK3/1 and AKT1. Small interfering RNA (siRNA) targeting human PPP3 catalytic subunit alpha (PPP3CA) was used to suppress PPP3CA protein expression in OFPAE cells. Compared with the scrambled siRNA, PPP3CA siRNA decreased PPP3CA protein levels by approximately 97% without altering protein levels of protein phosphatase 2 catalytic subunit alpha, total MAPK3/1, total AKT1, or glyceraldehyde-3-phosphate dehydrogenase. Knockdown of PPP3CA protein expression enhanced VEGF-stimulated, but not FGF2 stimulated, cell proliferation. Knockdown of PPP3CA protein expression did not significantly affect VEGF-induced MAPK3/1 and AKT1 phosphorylation but attenuated FGF2-induced MAPK3/1 and AKT1 phosphorylation. Thus, to our knowledge, the present study is the first to demonstrate successful knockdown of PPP3CA protein expression in any cell model using a single pair of double-strained siRNA. Moreover, specific knockdown of PPP3CA protein expression enhances VEGF stimulated, but not FGF2-stimulated, OFPAE cell proliferation and attenuates FGF2 induced, but not VEGF-induced, MAPK3/1 and AKT1 activation. Thus, PPP3CA differentially modulates the VEGF- and FGF2-stimulated cell proliferation and signaling cascades in OFPAE cells. These data also suggest that signaling molecules other than MAPK3/1 and AKT1 play an important role in VEGF- and FGF2 stimulated cell proliferation after knockdown of PPP3CA in OFPAE cells. PMID- 18509163 TI - HLA-G gene polymorphism in human placentas: possible association of G*0106 allele with preeclampsia and miscarriage. AB - Definite causes for several pathologies of pregnancy remain unknown. In light of several recent studies, however, diminished or aberrant HLA-G expression may be associated with certain complication of pregnancy and be linked to HLA-G polymorphism. We analyzed DNA from 60 normal placentas (controls), 140 placentas from miscarriage, 36 placentas from preeclampsia, 76 placentas from fetal hypotrophy, and 34 placentas with hypoxia for variations in coding regions (allelic groups G*0101 to G*0107) and the 14-bp deletion/insertion into the 3' untranslated region. No statistically significant differences were observed in the distribution of allelic group between pathological placentas and controls with the exception of G*0106 allele frequency in preeclamptic compared with control placentas (21.2% and 6.6%, respectively). A greater frequency of this allele also was observed in the two subgroups of miscarriage and hypoxia compared with that in controls. In addition, presence of the 14-bp sequence was prominent in preeclampsia compared with controls (60.8% vs. 35%, respectively), and homozygotes with deletion were not detected in the pathology. The results suggest that the G*0106 allele, which is coupled with the presence of the 14-bp sequence, contributes and/or is a relevant marker in some specific complications of pregnancy, especially preeclampsia. PMID- 18509164 TI - Cleavage of disulfide bonds in mouse spermatogenic cell-specific type 1 hexokinase isozyme is associated with increased hexokinase activity and initiation of sperm motility. AB - During epididymal transit, sperm acquire the ability to initiate rapid forward progressive motility on release into the female reproductive tract or physiological media. Glycolysis is the primary source of the ATP necessary for this motility in the mouse, and several novel glycolytic enzymes have been identified that are localized to the principal piece region of the flagellum. One of these is the spermatogenic cell-specific type 1 hexokinase isozyme (HK1S), the only member of the hexokinase enzyme family detected in sperm. Hexokinase activity was found to be lower in immotile sperm immediately after removal from the cauda epididymis (quiescent) than in sperm incubated in physiological medium for 5 min and showing rapid forward progressive motility (activated). However, incubating sperm in medium containing diamide, an inhibitor of disulfide bond reduction, resulted in lower motility and HK activity than in controls. HK1S was present in dimer and monomer forms in extracts of quiescent sperm but mainly as a monomer in motile sperm. A dimer-size band detected in quiescent sperm with phosphotyrosine antibody was not detected in activated sperm, and the monomer size band was enhanced. In addition, the general protein oxido-reductase thioredoxin-1 was able to catalyze the in vitro conversion of HK1S dimers to the monomeric form. These results strongly suggest that cleavage of disulfide bonds in HK1S dimers contributes to the increases in HK activity and motility that occur when mouse sperm become activated. PMID- 18509165 TI - Molecular identification and functional characterization of the kisspeptin/kisspeptin receptor system in lower vertebrates. AB - The KISS1 gene encodes the kisspeptin neuropeptide, which activates the KISS1 receptor (KISS1R; G protein-coupled receptor 54; GPR54) and participates in neuroendocrine regulation of GnRH secretion. To study the physiological function(s) and evolutionary conservation of KISS1, we cloned opossum, Xenopus, and zebrafish kiss1 cDNAs. Processing zebrafish, Xenopus, or opossum KISS proteins would liberate a carboxy-terminal amidated peptide with 52, 54, or 53 amino acid residues, respectively. Phylogenetic analysis of all known vertebrate KISS1 peptides showed clear clustering of the sequences according to canonical vertebrate classes. The zebrafish kiss1 gene consists of two exons and one intron. Real-time PCR analysis of two kiss1R cloned from zebrafish brain found expression of kiss1, kiss1ra, and kiss1rb, with kiss1ra-more similar to other piscine Kiss1 receptors-highly expressed in the gonads and kiss1rb in other nonbrain tissues. In females kiss1 mRNA levels gradually increased during the first few weeks of life to peak in fish with ovaries containing mature oocytes, while in males kiss1 mRNA levels peaked after 6 wk postfertilization when the testes exhibited initial stages of spermatogenesis and decreased after puberty. Zebrafish kiss1ra and kiss1rb were expressed differentially with similar patterns in both genders. These results indicate that the Kiss1/Kiss1r system may participate in puberty initiation in fish as well. Like human KISS1R, Kiss1ra transduces its activity via the PKC pathway, whereas Kiss1rb does so via both PKC and PKA pathways. The human KISS1R was highly activated by both huKISS10amide and zfKISS10amide, whereas both zebrafish Kiss1 receptor types were less sensitive to amidation. PMID- 18509166 TI - Molecular identification of lyso-glycerophosphocholines as endogenous immunosuppressives in bovine and rat gonadal fluids. AB - The ability of the gametes to escape detection by the immune system is vital to successful human reproduction. Furthermore, the observed capacity of the testis in some species to support tissue grafts without rejection (immunological privilege) indicates that spermatogenic cells are protected by local immunoregulatory mechanisms. One of these mechanisms involves targeting T cells for inactivation and destruction within the testicular environment. Although the fluids of the testis and ovary surrounding the developing gametes contain soluble factors that inhibit T cells, the identity of the molecule(s) responsible for this activity has been unknown. Using a specific T-cell proliferation assay to monitor bioactivity, these molecules were purified from bovine ovarian follicular fluid by methanol extraction and sequential reverse-phase HPLC (RP-HPLC). All purified active fractions coincided with the elution position on RP-HPLC of several small molecules ranging in size from 496 to 522 Da. The same molecules were localized to the immunosuppressive fractions of rat testicular interstitial fluid. The active molecules were identified, using capillary electrophoresis electrospray ionization mass spectroscopy, as lyso-glycerophosphocholines (lyso GPCs), namely, 1-palmitoyl-sn-glycero-3-phosphocholine, 1-oleoyl-sn-glycero-3 phosphocholine, a 18:2a/lyso-GPC (putatively, 1-linoleoyl-sn-glycero-3 phosphocholine), and a 20:4a/lyso-GPC (putatively, 1-arachidonyl-sn-glycero-3 phosphocholine). Comparison of the bioactivity and mass spectroscopy profiles of two of the purified molecules with their synthetic standards confirmed the identification. These molecules inhibit T-cell proliferation in response to activation and induce apoptosis of these cells in a time- and dose-dependent manner. The emergence of gonadal lyso-GPCs as potential regulators of critical immune events opens up new avenues of inquiry into the origins of autoimmune infertility and more generally into mechanisms of peripheral immunoregulation and the development of novel immunosuppressives. PMID- 18509167 TI - Journal honors Brian G. Till, MD, as recipient of the 2008 JCO Young Investigator Award. PMID- 18509168 TI - Hairy cell leukemia: treatment successes in the past 25 years. PMID- 18509169 TI - The oncology-rehabilitation interface: better systems needed. PMID- 18509170 TI - Fertility preservation: a vital survivorship issue for young women with breast cancer. PMID- 18509171 TI - Why learning to communicate with our patients is so important: using communication to enhance accrual to cancer clinical trials. PMID- 18509172 TI - Reversal of drug resistance in ovarian cancer: where do we go from here? PMID- 18509173 TI - Don Quixote and the quest for personalized medicine. PMID- 18509174 TI - Prevalence and treatment patterns of physical impairments in patients with metastatic breast cancer. AB - PURPOSE: Physical impairments cause profound functional declines in patients with cancer. Although common rehabilitation measures can address many impairments, the extent of their delivery is unknown. We studied these issues by quantifying physical impairments in patients with metastatic breast cancer and by assessing how they are addressed. PATIENTS AND METHODS: A consecutive sample of 163 community-dwelling patients with metastatic breast cancer was stratified by Karnofsky performance score and administered the Medical Outcomes Study Physical Function Subscale and the Older Americans Resource Study Activities of Daily Living subscales. Cancer-related physical impairments were identified through a physical examination, the 6-Minute Walk Test, and the Functional Independence Measure Mobility Subscale. Patients were questioned regarding the nature, type, and setting of treatments for impairments. Physical rehabilitation needs were determined through a consensus process involving physiatrists and physical/occupational therapists specializing in cancer. RESULTS: Ninety-two percent of patients (150 of 163) had at least one physical impairment. Among 530 identified impairments, 484 (92%) required a physical rehabilitation intervention and 469 (88%) required physical therapy (PT) and/or occupational therapy (OT). Only 30% of impairments requiring rehabilitation services and 21% of those requiring PT/OT received treatment. Impairments detected during hospitalization were overwhelmingly more likely to receive a rehabilitation intervention (odds ratio [OR] = 87.9; 95% CI, 28.5 to 271.4), and PT/OT (OR = 558.8; 95% CI, 187.0 to 1,669.6). Low socioeconomic and minority status were significantly associated with nontreatment. CONCLUSION: Remediable physical impairments were prevalent and poorly addressed among patients with metastatic breast cancer, drastically so in the outpatient setting. Undertreatment was particularly prominent among minority and socioeconomically disadvantaged groups. PMID- 18509175 TI - Safety of fertility preservation by ovarian stimulation with letrozole and gonadotropins in patients with breast cancer: a prospective controlled study. AB - PURPOSE: Because of the accompanying increase in estrogen levels, safety of performing in vitro fertilization (IVF) in women with breast cancer is unknown. Our goal was to determine the effect of controlled ovarian stimulation (COS) using a combination of letrozole with standard fertility medications on disease free survival in women undergoing embryo or oocyte cryopreservation before adjuvant chemotherapy. PATIENTS AND METHODS: A total of 215 women with breast cancer were prospectively evaluated for fertility preservation before adjuvant chemotherapy. Of those, 79 elected to undergo COS with letrozole and gonadotropins for embryo or oocyte cryopreservation. The remaining 136 patients underwent no fertility-preserving procedure and served as controls. RESULTS: Study and control groups were similar at enrollment except for a trend for higher estrogen-receptor positivity in the COS group (P = .08). Time between surgery and chemotherapy was longer for IVF patients (45.08 v 33.46 days; P < .01). Peak estradiol levels ranged from 58.4 to 1,166 pg/mL (mean, 405.94 +/- 256.64 pg/mL or 1,486.76 +/- 942.13 pmol/L) in COS patients. The median follow-up after chemotherapy was 23.4 months (range, 7.5 to 63.6 months) in the COS group and 33.05 months (range, 4.5 to 63.6) in the control group. The hazard ratio for recurrence after IVF was 0.56 (95% CI, 0.17 to 1.9), and the survival was not compromised compared with controls (P = .36). CONCLUSION: Ovarian stimulation with gonadotropins and letrozole for the purpose of fertility preservation is unlikely to cause substantially increased recurrence risk. Further research, including longer-term follow-up is needed to confirm these findings. PMID- 18509176 TI - Estrogen receptor expression and efficacy of docetaxel-containing adjuvant chemotherapy in patients with node-positive breast cancer: results from a pooled analysis. AB - PURPOSE: Several adjuvant chemotherapy trials suggested that cytotoxic treatment is less effective in patients with estrogen receptor (ER)-positive breast cancers. The aim of the present study was to assess the efficacy of adjuvant docetaxel and anthracycline therapy according to ER expression in two randomized clinical trials. PATIENTS AND METHODS: Pooled data from two randomized trials, BCIRG001 and PACS01, were examined. Hazard ratios for recurrence and survival were estimated by Cox proportional hazards models and were adjusted for clinical variables. Interaction between docetaxel and ER expression was tested. RESULTS: ER status was available for 3,329 patients (95% of all randomly assigned patients), of whom 75% (n = 2,493) were ER positive. Docetaxel therapy was associated with a 30% reduction in the risk of death (hazard ratio [HR] = 0.70; 95% CI, 0.54 to 0.91) in ER-positive patients and a 31% reduction (HR = 0.69; 95% CI, 0.52 to 0.94) in ER-negative patients. Docetaxel therapy was associated with a 21% reduction in the risk of recurrence (HR = 0.79; 95% CI, 0.66 to 0.93) in ER positive patients and a 31% reduction (HR = 0.69; 95% CI, 0.54 to 0.97) in ER negative patients. The interaction between docetaxel therapy and ER status was not statistically significant for either overall survival (P = .87) or disease free survival (P = .30). ER expression was also not predictive for docetaxel efficacy when it was analyzed as a semi-continuous variable based on percent of positive cells by immunohistochemistry (test for heterogeneity, P = .56 and .86 for overall survival and disease-free survival, respectively). CONCLUSION: In the pooled analysis of these two trials, docetaxel did not have a statistically significantly different effect on the risk of recurrence or death in ER-positive and ER-negative patients. PMID- 18509177 TI - Racial differences in clinical outcomes from metastatic breast cancer: a pooled analysis of CALGB 9342 and 9840--Cancer and Leukemia Group B. AB - PURPOSE: African American women are more likely to be diagnosed with metastatic breast cancer at the time of presentation than whites, and have shorter survival once diagnosed. This study examines racial differences in clinical outcomes in the setting of two large cooperative group randomized clinical trials. PATIENTS AND METHODS: The study cohort consisted of 787 white (80%) and 195 African American (20%) patients with metastatic breast cancer enrolled in two successive Cancer and Leukemia Group B (CALGB) trials using taxanes in the metastatic setting. Differences in overall survival (OS), response incidence, and time to treatment failure (TTF) were examined by race. In addition, differences in the incidence of baseline and treatment-related toxicities were examined. RESULTS: With 779 deaths (166 African Americans and 613 whites), median OS was 14.3 months for African Americans and 18.75 months for whites (hazard ratio [HR] = 1.37; 95% CI, 1.15 to 1.63). When adjusted for prognostic factors, African Americans had a 24% increase in the hazard of death compared with whites (HR = 1.24; 95% CI, 1.02 to 1.51). No significant differences in TTF or overall response to therapy were seen. No clinically significant toxicity differences were seen. CONCLUSION: African Americans with metastatic breast cancer have an increased hazard of death compared with whites despite the receipt of similar per-protocol treatment, but experience no differences in TTF or overall response to therapy. We hypothesize that more direct and robust measures of comorbidities, and perhaps other factors such as receipt of subsequent therapy could help further explain the observed survival difference. PMID- 18509178 TI - Influence of clinical communication on patients' decision making on participation in clinical trials. AB - PURPOSE: To investigate how communication among physicians, patients, and family/companions influences patients' decision making about participation in clinical trials. PATIENTS AND METHODS: We video recorded 235 outpatient interactions occurring among oncologists, patients, and family/companions (if present) at two comprehensive cancer centers. We combined interaction analysis of the real-time video-recorded observations (collected at Time 1) with patient self reports (Time 2) to determine how communication about trial offers influenced accrual decisions. RESULTS: Clinical trials were explicitly offered in 20% of the interactions. When offers were made and patients perceived they were offered a trial, 75% of patients assented. Observed messages (at Time 1) directly related to patients' self-reports regarding their decisions (2 weeks later), and how they felt about their decisions and their physicians. Specifically, messages that help build a sense of an alliance (among all parties, including the family/companions), provide support (tangible assistance and reassurance about managing adverse effects), and provide medical content in language that patients and family/companions understand are associated with the patient's decision and decision-making process. CONCLUSION: In two urban, National Cancer Institute designated comprehensive cancer centers, a large percentage of patients are not offered trials. When offered a trial, most patients enroll. The quality and quantity of communication occurring among the oncologist, patient, and family/companion when trials are discussed matter in the patient's decision making process. These findings can help increase physician awareness of the ways that messages and communication behaviors can be observed and evaluated to improve clinical practice and research. PMID- 18509179 TI - Phase III study of valspodar (PSC 833) combined with paclitaxel and carboplatin compared with paclitaxel and carboplatin alone in patients with stage IV or suboptimally debulked stage III epithelial ovarian cancer or primary peritoneal cancer. AB - PURPOSE: To compare the safety and efficacy of carboplatin and paclitaxel administered with or without the multidrug resistance modulator valspodar (PSC 833) in untreated patients with advanced ovarian or primary peritoneal cancer. PATIENTS AND METHODS: Seven hundred sixty-two patients with stage IV or suboptimally debulked stage III ovarian or primary peritoneal cancer were randomly assigned to receive either valspodar 5 mg/kg every 6 hours for 12 doses, paclitaxel 80 mg/m(2), and carboplatin area under the curve (AUC) 6 (PC-PSC; n = 381) or paclitaxel 175 mg/m(2) and carboplatin AUC 6 (PC; n = 381). Time to disease progression (TTP) was the primary end point. Secondary end points were overall survival time (OS), response rate (RR), safety, and tolerability. RESULTS: With a median follow-up of 736 days (range, 1 to 2,280 days), the median TTP was 13.2 and 13.5 months in the PC-PSC and PC groups, respectively (P = .67); the median OS was 32 and 28.9 months, respectively (P = .94). The overall RR was higher in the PC group (41.5% v 33.6%; P = .02). Central and peripheral nervous system and GI toxicities were more common in the PC-PSC group. Ataxia occurred in 53.5% and 3.2% of PC-PSC-and PC-treated patients, respectively. Febrile neutropenia occurred more frequently in the PC-PSC group. More PC-PSC-treated patients discontinued therapy because of adverse events (AEs), experienced serious AEs, and required paclitaxel dose reductions. CONCLUSION: The addition of valspodar to PC did not improve TTP or OS and was more toxic compared with PC in untreated patients with advanced ovarian or primary peritoneal cancer. PMID- 18509180 TI - Incidence of venous thromboembolism in patients with ovarian cancer undergoing platinum/paclitaxel-containing first-line chemotherapy: an exploratory analysis by the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group. AB - PURPOSE: Venous thromboembolism (VTE) has been associated with negative prognosis in cancer patients. Most series reporting on VTE have included different tumor types not differentiating between recurrent or primary disease. Data regarding the actual impact of VTE on primary advanced ovarian cancer (AOC) are limited. PATIENTS AND METHODS: Between 1995 and 2002, the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study group (AGO-OVAR) recruited 2,743 patients with AOC in three prospectively randomized trials on platinum paclitaxel based chemotherapy after primary surgery. Pooled data analysis was performed to evaluate incidence, predictors, and prognostic impact of VTE in AOC. Survival curves were calculated for the VTE incidence. Univariate analysis and Cox regression analysis were performed to identify independent predictors of VTE and mortality. RESULTS: Seventy-six VTE episodes were identified, which occurred during six to 11 cycles of adjuvant chemotherapy; 50% of them occurred within 2 months postoperatively. Multivariate analysis identified body mass index higher than 30 kg/m(2) and increasing age as independent predictors of VTE. International Federation of Gynecology and Obstetrics stage and surgical radicality did not affect incidence. Overall survival was significantly reduced in patients with VTE (median, 29.8 v 36.2 months; P = .03). Multivariate analysis identified pulmonary embolism (PE), but not deep vein thrombosis alone, to be of prognostic significance. In addition, VTE was not identified to significantly affect progression-free survival. CONCLUSION: Patients with AOC have their highest VTE risk within the first 2 months after radical surgery. Only VTE complicated by symptomatic PE have been identified to have a negative impact on survival. Studies evaluating the role of prophylactic anticoagulation during this high risk postoperative period are warranted. PMID- 18509181 TI - Predictive biomarkers of chemotherapy efficacy in colorectal cancer: results from the UK MRC FOCUS trial. AB - PURPOSE: Candidate predictive biomarkers for irinotecan and oxaliplatin were assessed in 1,628 patients in Fluorouracil, Oxaliplatin, CPT-11: Use and Sequencing (FOCUS), a large randomized trial of fluorouracil alone compared with fluorouracil and irinotecan and compared with fluorouracil and oxaliplatin in advanced colorectal cancer. METHODS: The candidate biomarkers were: tumor immunohistochemistry for MLH1/MSH2, p53, topoisomerase-1 (Topo1), excision repair cross-complementing gene 1 (ERCC1), O-6-methylguanine-DNA-methyltranserase (MGMT), and cyclooxygenase 2 (COX2); germline DNA polymorphisms in GSTP1, ABCB1, XRCC1, ERCC2, and UGT1A1. These were screened in more than 750 patients for interaction with benefit from irinotecan or oxaliplatin; two markers (Topo1 and MLH1/MSH2) met criteria to be taken forward for analysis in the full population. Primary end points were progression-free survival (PFS) and overall survival. RESULTS: One thousand three hundred thirteen patients (81%) were assessable for Topo1 immunohistochemistry (low, < 10%; moderate, 10% to 50%; or high, > 50% tumor nuclei). In patients with low Topo1, PFS was not improved by the addition of either irinotecan (hazard ratio [HR], 0.98; 95% CI, 0.78 to 1.22) or oxaliplatin (HR, 0.85; 95% CI, 0.68 to 1.07); conversely, patients with moderate/high Topo1 benefited from the addition of either drug (HR, 0.48 to 0.70 in all categories; interaction P = .005; overall, P = .001 for irinotecan; P = .05 for oxaliplatin). High Topo1 was associated with a major overall survival benefit with first-line combination chemotherapy (HR, 0.60; median benefit, 5.3 months); patients with moderate or low Topo1 did not benefit (HR, 0.92 and 1.09, respectively; interaction P = .005). MLH1/MSH2 did not show significant interaction with treatment, although the low rate of loss (4.4%) limits the power of the study for this biomarker. CONCLUSION: Topo1 immunohistochemistry identified subpopulations that did or did not benefit from irinotecan, and possibly also from oxaliplatin. If verified independently, this information will contribute to the individualization of treatment for colorectal cancer. PMID- 18509182 TI - Phase I trial of the human immunodeficiency virus protease inhibitor nelfinavir and chemoradiation for locally advanced pancreatic cancer. AB - PURPOSE: Preclinically, HIV protease inhibitors radiosensitize tumors with activated PI3-kinase/Akt pathway. We determined the toxicity of nelfinavir chemoradiotherapy in borderline resectable and unresectable pancreatic cancer. PATIENTS AND METHODS: Oral nelfinavir (2 x 1,250 mg) was started 3 days before and continued throughout chemoradiotherapy to 50.4 Gy (boost, 59.4 Gy) in 12 patients. Two gemcitabine dose levels (DL) were tested (200 mg/m(2) and 300 mg/m(2) on days 1, 8, 22, and 29). Cisplatin was administered on the same days at 30 mg/m(2). Phospho-Akt downregulation by nelfinavir was monitored by immunoblotting in patient leukocytes. Restaging positron emission tomography (PET)/computed tomography (CT) and CA19-9 levels served to assess response, and responding tumors were resected. RESULTS: At each DL, five of six patients completed chemoradiotherapy, and two of 12 patients had incomplete chemoradiotherapy because of clinical depression (DL1) and peritoneal metastasis (DL2). Grade 4 toxicities were a transaminase elevation (DL2) as a result of biliary stent occlusion and acute cholecystitis as a result of peritoneal metastasis (DL2). Stent occlusions led to dose-limiting toxicities of grade 3 liver enzyme and bilirubin elevations (two patients at DL1, one patient at DL2). Grade 3 nausea and vomiting occurred in a DL2 patient, and weight loss occurred in a DL1 patient who refused supportive feeding. Secondary complete resection was possible in six of 10 patients with complete chemoradiotherapy, including one tumor with pathologic sterilization. Partial CT responses were observed in five of 10 patients who completed chemoradiotherapy. Of nine patients assessable by PET,responses were complete in five patients and partial patients, and stable disease was observed in two patients. CONCLUSION: The combination of nelfinavir and chemoradiotherapy showed acceptable toxicity and promising activity in patients with pancreatic cancer. PMID- 18509183 TI - High expression of macrophage colony-stimulating factor in peritumoral liver tissue is associated with poor survival after curative resection of hepatocellular carcinoma. AB - PURPOSE: To investigate prognostic values of the intratumoral and peritumoral expression of macrophage colony-stimulating factors (M-CSF) in hepatocellular carcinoma (HCC) patients after curative resection. PATIENTS AND METHODS: Expression of M-CSF and density of macrophages (M Phi) were assessed by immunohistochemistry in tissue microarrays containing paired tumor and peritumoral liver tissue from 105 patients who had undergone hepatectomy for histologically proven HCC. Prognostic value of these and other clinicopathologic factors was evaluated. RESULTS: Neither intratumoral M-CSF nor M Phi density was associated with overall survival (OS) or disease-free survival (DFS). High peritumoral M-CSF and M Phi density, which correlated with large tumor size, presence of intrahepatic metastasis, and high TNM stage, were independent prognostic factors for both OS (P = .001 and P < .001, respectively) and DFS (P = .001 and P = .003, respectively) and affected incidence of early recurrence. In a small HCC subset, peritumoral M-CSF was also correlated with both OS and DFS (P = .038 and P = .001, respectively). The combination of peritumoral M-CSF and M Phi had a better power to predict the patients' death and disease recurrence (P < .001 for both). CONCLUSION: High peritumoral M-CSF and M Phi were associated with HCC progression, disease recurrence, and poor survival after hepatectomy, highlighting the importance of peritumoral tissue in the recurrence and metastasis of HCC. M-CSF and M Phi may be targets of postoperative adjuvant therapy. PMID- 18509184 TI - Specific EGFR mutations predict treatment outcome of stage IIIB/IV patients with chemotherapy-naive non-small-cell lung cancer receiving first-line gefitinib monotherapy. AB - PURPOSE: To explore predictive factors for time to treatment failure (TTF) in chemotherapy-naive non-small-cell lung cancer (NSCLC) patients receiving gefitinib treatment. PATIENTS AND METHODS: We designed a phase II study to test gefitinib antitumor efficacy in advanced-stage, chemotherapy-naive NSCLC patients. Patients were treated with gefitinib 250 mg/d. Tumor assessments were performed every 2 months. Responding or stable patients were treated until progression or unacceptable toxicity. All scans were reviewed independently. EGFR exons 18-21 sequence, K-ras exon 2 sequence, and MET gene copy numbers were examined in available samples. Clinical or molecular predictors of TTF were examined by multivariate analysis. RESULTS: One hundred six patients were enrolled. Ninety patients had tumor samples for biomarker tests. Overall response rate was 50.9% (95% CI, 41.4% to 60.4%). Median TTF was 5.5 months, and median overall survival (OS) was 22.4 months. The response rate and median TTF of the patients with exon 19 deletion (n = 20) were 95.0% and 8.9 months, for exon 21 L858R mutation (n = 23) were 73.9% and 9.1 month, and for other types of EGFR mutations (N = 12) were 16.7% and 2.3 months, respectively. In multivariate analysis, the presence of EGFR deletion exon 19 or L858R EGFR mutations in adenocarcinoma patients predicted longer TTF. High copy number of MET seemed to correlate with shorter TTF in patients with gefitinib-sensitive activating EGFR mutations. CONCLUSION: In this prospective study, EGFR exon 19 deletion or L858R mutations in adenocarcinoma were the best predictors for longer TTF in stage IIIB/IV chemotherapy-naive NSCLC patients receiving first-line gefitinib monotherapy. PMID- 18509185 TI - Quality of life scores predict survival among patients with head and neck cancer. AB - PURPOSE: The purpose of this study was to examine whether quality of life (QOL) scores predict survival among patients with head and neck cancer, controlling for demographic, health behavior, and clinical variables. PATIENTS AND METHODS: A self-administered questionnaire was given to 495 patients being treated for head and neck cancer while they were waiting to be seen for a clinic appointment. Data collected from the survey included demographics, health behaviors, and QOL as measured by Short Form-36 (SF-36) physical and mental component scores and the Head and Neck QOL scores. Clinical measures were collected by chart abstraction. Kaplan-Meier plots and univariate and multivariate Cox proportional hazards models were used to determine the association between QOL scores and survival time. RESULTS: After controlling for age, time since diagnosis, marital status, education, tumor site and stage, comorbidities, and smoking, the SF-36 physical component score and three of the four Head and Neck QOL scales (pain, eating, and speech domains) were associated with survival. Controlling for the same variables, the SF-36 mental component score and the emotional domain of the Head and Neck QOL were not associated with survival. CONCLUSION: QOL instruments may be valuable screening tools to identify patients who are at high risk for poor survival. Those with low QOL scores could be followed more closely, with the potential to identify recurrence earlier and perform salvage treatments, thereby possibly improving survival for this group of patients. PMID- 18509186 TI - Guidelines for the management of pediatric and adult tumor lysis syndrome: an evidence-based review. AB - PURPOSE: Tumor lysis syndrome (TLS) has recently been subclassified into either laboratory TLS or clinical TLS, and a grading system has been established. Standardized guidelines, however, are needed to aid in the stratification of patients according to risk and to establish prophylaxis and treatment recommendations for patients at risk or with established TLS. METHODS: A panel of experts in pediatric and adult hematologic malignancies and TLS was assembled to develop recommendations and guidelines for TLS based on clinical evidence and standards of care. A review of relevant literature was also used. RESULTS: New guidelines are presented regarding the prevention and management of patients at risk of developing TLS. The best management of TLS is prevention. Prevention strategies include hydration and prophylactic rasburicase in high-risk patients, hydration plus allopurinol or rasburicase for intermediate-risk patients, and close monitoring for low-risk patients. Primary management of established TLS involves similar recommendations, with the addition of aggressive hydration and diuresis, plus allopurinol or rasburicase for hyperuricemia. Alkalinization is not recommended. Although guidelines for rasburicase use in adults are provided, this agent is currently only approved for use in pediatric patients in the United States. CONCLUSION: The potential severity of complications resulting from TLS requires measures for prevention in high-risk patients and prompts treatment in the event that symptoms arise. Recognition of risk factors, monitoring of at-risk patients, and appropriate interventions are the key to preventing or managing TLS. These guidelines should assist in the prevention of TLS and improve the management of patients with established TLS. PMID- 18509187 TI - Fatal toxic epidermal necrolysis associated with cetuximab in a patient with colon cancer. PMID- 18509188 TI - Strumal carcinoid tumor of the ovary. PMID- 18509189 TI - Pseudohyperkalemia in chronic lymphocytic leukemia. PMID- 18509190 TI - Swimming upstream: an advocate reflects on cancer research and social realities. PMID- 18509191 TI - Stopping trials for benefit can (sometimes) benefit patients. PMID- 18509192 TI - Hepatic arterial infusion chemotherapy for metastases from colorectal cancer: is it really the end of an era? PMID- 18509193 TI - The Koebner-phenomenon in epidermal growth factor receptor inhibitor-induced cutaneous adverse effects. PMID- 18509194 TI - Sphingomyelinase acts by an area-activated mechanism on the liquid-expanded phase of sphingomyelin monolayers. AB - We describe the localization of Alexa-488-labeled SMase in SM/ceramide (Cer) lipid monolayers containing segregated liquid-condensed (LC) Cer-enriched domains surrounded by a continuous liquid-expanded (LE) SM-enriched phase. Langmuir Schaefer films were made in order to visualize the labeled enzyme. Independently of initial conditions Alexa-SMase is preferably localized in the SM-enriched LE phase and it is not enriched at the domain boundaries. A novel mechanism is proposed for the action of SMase, which can also explain the regulatory effect of the surface topography on the enzyme activity. The homogeneous enzymatic generation of Cer in the LE phase leads to a meta-stable, kinetically trapped, supersaturated mixed monolayer. This effect acts as driving force for the segregation of the Cer-enriched domain following classical nucleation mechanisms. Accordingly, the number and size of Cer-enriched domains are determined by the extent of Cer supersaturation in the LE phase rather than by the SMase local activity. The kinetic barrier for nucleation, for which a compositional gap of at least 53 mol% of Cer is necessary to reach a thermodynamically stable LC phase, can explain the lag time to reaching full catalytic activity. Altogether, the data support an "area-activated mechanism," in which the enzyme is homogeneously active over the LE surface. PMID- 18509195 TI - Expression of CETP and of splice variants induces the same level of ER stress despite secretion efficiency differences. AB - The cholesteryl ester transfer protein (CETP) gene has been associated with a variety of phenotypes, including HDL-cholesterol levels and, more sporadically, with cardiovascular disease, obesity, and extreme longevity. Alterations of CETP activity levels can be caused by single-base polymorphisms as well as by alternative splicing. In addition to the previously characterized alternative splicing that skips exon 9, we found additional minor variants and characterized the activity of the resultant proteins. The novel variants skipped exon 9 sequences and inserted one of two in-frame exons from Alu-derived intronic sequences. None of the alternatively spliced variants are efficiently secreted, and coexpression of them inhibits wild-type CETP secretion. Expression of the alternative spliced variants causes an induction of genes linked to the endoplasmic reticulum (ER) stress response, including the neighboring HERPUD1 (homocysteine- and ER stress-inducible protein, ubiquitin-like domain-containing) gene. Unexpectedly, even though wild-type CETP is secreted much more efficiently than spliced variants, it induces the same degree of stress response as spliced variants, whereas a control secreted protein does not. CETP plays a complex role in modulating ER stress, with its expression inducing the response and its cholesteryl ester transfer activity and differential splicing modulating the response in other ways. PMID- 18509196 TI - Liver X receptor-mediated activation of reverse cholesterol transport from macrophages to feces in vivo requires ABCG5/G8. AB - Liver X receptor (LXR) agonists increase both total fecal sterol excretion and macrophage-specific reverse cholesterol transport (RCT) in vivo. In this study, we assessed the effects of ABCG5/G8 deficiency as well as those of LXR agonist induction of RCT from macrophages to feces in vivo. A [(3)H]cholesterol-labeled macrophage cell line was injected intraperitoneally into ABCG5/G8-deficient (G5/G8(-/-)), heterozygous (G5G8(+/-)), and wild-type G5/G8(+/+) mice. G5/G8(-/ )mice presented increased radiolabeled HDL-bound [(3)H]cholesterol 24 h after the label injection. However, the magnitude of macrophage-derived [(3)H]cholesterol in liver and feces did not differ between groups. A separate experiment was conducted in G5G8(+/+) and G5G8(-/-) mice treated with or without the LXR agonist T0901317. Treatment with T0901317 increased liver ABCG5/G8 expression, which was associated with a 2-fold increase in macrophage-derived [(3)H]cholesterol in feces of G5/G8(+/+) mice. However, T0901317 treatment had no effect on fecal [(3)H]cholesterol excretion in G5G8(-/-) mice. Additionally, LXR activation stimulated the fecal excretion of labeled cholesterol after an intravenous injection of HDL-[(3)H]cholesteryl oleate in G5/G8(+/+) mice, but failed to enhance fecal [(3)H]cholesterol in G5/G8(-/-) mice. Our data provide direct in vivo evidence of the crucial role of ABCG5 and ABCG8 in LXR-mediated induction of macrophage-specific RCT. PMID- 18509197 TI - Composition of adipose tissue and marrow fat in humans by 1H NMR at 7 Tesla. AB - Proton NMR spectroscopy at 7 Tesla (7T) was evaluated as a new method to quantify human fat composition noninvasively. In validation experiments, the composition of a known mixture of triolein, tristearin, and trilinolein agreed well with measurements by (1)H NMR spectroscopy. Triglycerides in calf subcutaneous tissue and tibial bone marrow were examined in 20 healthy subjects by (1)H spectroscopy. Ten well-resolved proton resonances from triglycerides were detected using stimulated echo acquisition mode sequence and small voxel ( approximately 0.1 ml), and T(1) and T(2) were measured. Triglyceride composition was not different between calf subcutaneous adipose tissue and tibial marrow for a given subject, and its variation among subjects, as a result of diet and genetic differences, fell in a narrow range. After correction for differential relaxation effects, the marrow fat composition was 29.1 +/- 3.5% saturated, 46.4 +/- 4.8% monounsaturated, and 24.5 +/- 3.1% diunsaturated, compared with adipose fat composition, 27.1 +/- 4.2% saturated, 49.6 +/- 5.7% monounsaturated, and 23.4 +/- 3.9% diunsaturated. Proton spectroscopy at 7T offers a simple, fast, noninvasive, and painless method for obtaining detailed information about lipid composition in humans, and the sensitivity and resolution of the method may facilitate longitudinal monitoring of changes in lipid composition in response to diet, exercise, and disease. PMID- 18509198 TI - Factors associated with hospital retention of RNs in the New York City Metropolitan Area: an analysis of the 1996, 2000, and 2004 National Sample Survey of Registered Nurses. AB - The nursing shortage is well documented, and government estimates indicate that shortfalls will worsen in the future. As the largest employer of registered nurses (RNs), hospitals are the most seriously affected by shortages, as they compete with other employment settings for limited nursing resources. Recruitment remains the primary avenue for ensuring staffing levels, but retention is increasingly important as applicant pools shrink because of demographic and employment trends. Effective retention strategies must address the factors that contribute to exodus of RNs from hospitals, as well as isolating the factors that enable RNs to remain in hospital employment. This secondary analysis of the 1996, 2000, and 2004 National Sample Survey of Registered Nurses examines the demographic, employment, and educational factors associated with working in hospitals, having full-time status, and holding patient care positions. The findings suggest that hospitals must address nonwork issues to retain nursing personnel. Relevant policy issues are examined and strategies for effective retention are offered. PMID- 18509199 TI - The role of the bone marrow in neutrophil clearance under homeostatic conditions in the mouse. AB - In humans, 10(11) neutrophils are released from the bone marrow per day, and these cells have a half-life in the blood of only approximately 6.5 h. Although it is generally believed that neutrophils are cleared from the circulation via the liver and spleen, in this study using (111)In-labeled senescent neutrophils, we show that in mice, 32% of neutrophils are cleared from the circulation via the bone marrow. We have previously shown that senescent neutrophils home to the bone marrow in a CXCR4-dependent manner, and we show here that pretreatment of neutrophils with pertussis toxin significantly inhibits neutrophil clearance via the bone marrow (75%), consistent with a role for chemokines in this process. By labeling senescent neutrophils with inert fluorescent microspheres, we have tracked their fate and shown that in vivo, they are ultimately phagocytosed by bone marrow stromal macrophages. Finally, we show that under noninflammatory conditions, circulating levels of neutrophils are regulated by granulocyte-colony stimulating factor (G-CSF), but not interleukin-17. Interestingly, we report that the uptake of apoptotic neutrophils by bone marrow macrophages stimulates their production of G-CSF in vitro. Taken together, these data provide evidence that the bone marrow represents a major site of neutrophil clearance in mice. PMID- 18509200 TI - Intermediate filament protein synemin contributes to the migratory properties of astrocytoma cells by influencing the dynamics of the actin cytoskeleton. AB - We have shown previously that, in astrocytoma cells, synemin is present at the leading edge, an unusual localization for an intermediate filament (IF) protein. Here, we report that synemin down-regulation with specific small hairpin RNAs (shRNAs) sharply decreased the migration of astrocytoma cells. The presence of synemin at the leading edge also correlated with a high migratory potential, as shown by comparing astrocytoma cells to carcinoma cells without synemin at the leading edge. Synemin-silenced astrocytoma cells were smaller and spread more slowly than controls. In addition, synemin silencing reduced proliferation without increasing apoptosis. The adhesion to substratum and distribution of vinculin in focal contacts of synemin-silenced astrocytoma cells were similar to those of controls. Synemin-silenced cells, however, exhibited a reduction in the amount of filamentous (F) -actin and of alpha-actinin, but not of vinculin, associated with F-actin. Altogether, these results demonstrate that synemin is important for the malignant behavior of astrocytoma cells and that it contributes to the high motility of these cells by modulating the dynamics of alpha-actinin and actin. PMID- 18509201 TI - Overexpression of Dyrk1A contributes to neurofibrillary degeneration in Down syndrome. AB - Adults with Down syndrome (DS) develop Alzheimer neurofibrillary degeneration in the brain, but the underlying molecular mechanism is unknown. Here, we report that the presence of an extra copy of the dual-specificity tyrosine phosphorylated and regulated kinase 1A (Dyrk1A) gene due to trisomy 21 resulted in overexpression of Dyrk1A and elevated kinase activity in DS brain. Dyrk1A phosphorylated tau at several sites, and these sites were hyperphosphorylated in adult DS brains. Phosphorylation of tau by Dyrk1A primed its further phosphorylation by glycogen synthase kinase-3beta (GSK-3beta). Dyrk1A-induced tau phosphorylation inhibited tau's biological activity and promoted its self aggregation. In Ts65Dn mouse brain, an extra copy of the Dyrk1A gene caused increased expression and activity of Dyrk1A and resulted in increased tau phosphorylation. These findings strongly suggest a novel mechanism by which the overexpression of Dyrk1A in DS brain causes neurofibrillary degeneration via hyperphosphorylating tau. PMID- 18509202 TI - Progression of cardiovascular autonomic dysfunction in patients with type 2 diabetes: a 7-year follow-up study. AB - OBJECTIVE: We investigated whether cardiovascular autonomic dysfunction was associated with glycemic control status over time in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: From 1999 to 2000, cardiovascular autonomic nerve function testing (AFT) was performed on patients with type 2 diabetes (n = 1,021) and was followed-up in 2006 and February 2008. Tests for cardiovascular autonomic functions measured heart rate variability parameters (expiration-to-inspiration [E/I] ratio, responses to the Valsalva maneuver, and standing). AFT scores were determined from the results of the each test as follows: 0 for normal and 1 for abnormal. We began with those who had a score of 0 and assessed the changes in total score along with biannual A1C levels. RESULTS: At follow-up, the development of cardiovascular autonomic dysfunction was 34.5% (E/I ratio 21.9%, Valsalva maneuver 77.8%, and posture 58.9%; n = 783). The development of cardiovascular autonomic dysfunction was higher in older patients (P < 0.001); in those with longer duration of diabetes (P < 0.001); of hypertension (P = 0.005), and of diabetic retinopathy (P < 0.001); and in those who had higher levels of microalbuminuria (P = 0.002). Logistic regression analysis revealed that the development of cardiovascular autonomic dysfunction was strongly associated with the mean A1C level during the follow-up period (mean A1C >9.0% vs. or=6.1 mmol/l), the mean number of foot problems for those with insensate feet was 3 compared with 0.3 among those with sensation (P < 0.001). In multivariate logistic regression female sex, low education, A1C, smoking, and homocysteine were independently associated with neuropathy, after controls for age. CONCLUSIONS: Neuropathy prevalence is high, given the young age of our participants (mean 40 years) and was present among those with undiagnosed diabetes. The high number and type of foot problems places this population at increased risk for ulceration; the low level of foot care in the community increases the risk. Homocysteine is a risk factor that may be related to lifestyle and requires further investigation. PMID- 18509209 TI - Explaining the decline in early mortality in men and women with type 2 diabetes: a population-based cohort study. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that changing utilization of lipid-lowering, antihypertensive, and oral hypoglycemic drugs may be associated with trends in all-cause mortality in men and women with type 2 diabetes. RESEARCH DESIGN AND METHODS: This was a cohort study in 197 general practices in the U.K. General Practice Research Database including 48,579 subjects with type 2 diabetes first diagnosed between 1996 and 2006. Measures included all-cause mortality and prescription of hypoglycemic, lipid-lowering, and antihypertensive drugs. RESULTS: From 1996 to 2006, incidence of type 2 diabetes increased and the mean age at diagnosis declined in women. Prescription of statins within 12 months of diagnosis increased (1996, women 4.9%, men 5.1%; 2005, women 63.5%, men 71.0%), as did drugs acting on the renin-angiotensin system (1996, women 19.4%, men 21.5%; 2005, women 45.5%, men 54.6%) and metformin (1996, women 19.1%, men 15.8%; 2005, women 45.5%, men 42.8%), whereas prescription of sulfonylureas declined. All-cause mortality in the first 24 months after diabetes diagnosis declined in men from 47.9 per 1,000 person-years for subjects with diabetes diagnosed in 1996 to 25.2 for subjects with diabetes diagnosed in 2006 and in women from 37.4 in 1996 to 27.6 in 2006. In a multiple regression model adjusting for age and comorbidity, prescription of statins before or after diagnosis, renin-angiotensin system drugs before or after diagnosis, and metformin after diagnosis were associated with lower mortality. CONCLUSIONS: Widespread implementation of more effective prescribing to control lipids, blood glucose, and blood pressure may have contributed to recent declines in early mortality in men and women with type 2 diabetes. PMID- 18509210 TI - Diabetes complications in youth: qualitative analysis of parents' perspectives of family learning and knowledge. AB - OBJECTIVE: Youth with type 1 diabetes face long-term risks of health complications of the disease. Little is known about patients' and parents' knowledge, acquisition of information, and family communication regarding these complications. This paper reports qualitative analyses of parental focus-group discussions of this topic. RESEARCH DESIGN AND METHODS: A total of 47 participants (30 mothers, 14 fathers, and 3 others) representing 33 children between the ages of 8 and 18 years with type 1 diabetes participated in 1 of 13 focus groups. Open-ended questions focused on the type and amount of information about long-term complications presented to parents by health care professionals at different time points, as well as the way that information was presented. Questions also elicited details about parent-child communication and exposure to misconceptions about diabetes complications. RESULTS: Qualitative analysis of the transcribed focus groups revealed that participants experienced significant anxiety about diabetes complications, with a shift from concern about daily management tasks to concern about long-term complications over time. Participants desired a flexible, collaborative educational approach, especially regarding the timing and type of information, relative to the child's age and duration of diabetes. Many parents wanted more sensitive communication and emotional support from health care providers. Motivating children appeared to be a particular challenge; family burnout with regard to diabetes care over time was reported. Knowledge was gained in many ways, yet misinformation was uncommon. CONCLUSIONS: Obtaining information about long-term complications is an important process that changes over the course of the disease and with the child's developmental level. More research is needed, especially regarding youth knowledge, learning, and beliefs about diabetes complications. PMID- 18509211 TI - Mitigating case mix factors by choice of glycemic control performance measure threshold. AB - OBJECTIVE: Performance measures are tools for assessing quality of care but may be influenced by patient factors. We investigated how currently endorsed performance measures for glycemic control in diabetes may be influenced by case mix composition. We assessed differences in A1C performance measure threshold attainment by case mix factors for A1C >9% and examined how lowering the threshold to A1C >8% or >7% changed these differences. RESEARCH DESIGN AND METHODS: Using data from the 1999-2002 National Health and Nutrition Examination Survey for 843 adults self-reporting diabetes, we computed the mean difference in A1C threshold attainment of >9, >8, and >7% by various case mix factors. The mean difference is the average percentage point difference in threshold attainment for population groups compared with that for the overall population. RESULTS: Diabetes medication was the only factor for which the difference in threshold attainment increased at lower thresholds, with mean differences of 5.7 percentage points at A1C >9% (reference), 10.1 percentage points at A1C >8% (P < 0.05), and 14.1 percentage points at A1C >7% (P < 0.001). CONCLUSIONS: As 87% of U.S. adults have A1C <9%, a performance measure threshold of >9% will not drive major improvements in glycemic control. Lower thresholds do not exacerbate differences in threshold attainment for most factors. Reporting by diabetes medication use may compensate for heterogeneous case mix when a performance measure threshold of A1C >8% or lower is used. PMID- 18509212 TI - Obesity and undiagnosed diabetes in the U.S. AB - OBJECTIVE: To study whether obese individuals, who are at higher risk for diabetes and disparities in care than nonobese individuals, are more likely to have undiagnosed diabetes. RESEARCH DESIGN AND METHODS: We performed an analysis of 5,514 adult participants in the 1999-2004 National Health and Nutrition Examination Survey. Participants were interviewed about sociodemographic and medical data, including whether they had been diagnosed with diabetes, and were examined for height, weight, and fasting plasma glucose level >or=126 mg/dl or by previous physician diagnosis. After categorizing participants into normal weight, overweight, and obese according to BMI, the prevalence and diagnosis of diabetes across BMI categories was compared using chi(2). RESULTS: Of the 9.8% (weighted sample) of participants who had diabetes, based on fasting glucose levels and self-reported diagnosis, 28.1% were undiagnosed, translating to an estimated 5.2 million people in the U.S. population. The proportion undiagnosed was not significantly different among normal-weight (22.2%), overweight (32.5%), or obese adults (27.4%). Nevertheless, obese adults comprise more than half of the undiagnosed diabetes cases (2.7 million). Relative to normal-weight adults, the adjusted odds ratio (OR) for having undiagnosed diabetes was 1.50 (0.73-3.08) in overweight and 1.37 (0.72-2.63) in obese adults. CONCLUSIONS: Despite a higher underlying risk of diabetes and widespread clinical recognition of this higher risk, obesity does not increase the likelihood that an individual's diabetes will be diagnosed. PMID- 18509213 TI - Caring for children with mental disorders: do state parity laws increase access to treatment? AB - BACKGROUND: High prevalence rates of mental health disorders in childhood have garnered increased public attention in recent years. Yet, among children diagnosed with serious mental health problems, a majority receive no treatment. Improving access to mental health services for children with behavioral and emotional disorders constitutes an important policy concern. AIMS OF THE STUDY: To study whether living in a state that has implemented a mental health parity law affects a child's use of outpatient mental health services. METHODS: We use the National Survey of America's Families (NSAF) 1997 to 2002 to study whether enacting a state parity law increases the probability that a privately insured child receives mental health treatment (N=26,916). Using a differences-in differences model, we control for detailed information on a child's health and functioning, and compare mental health care use before and after state parity implementation with non-parity states serving as a comparison group. RESULTS: Regression results indicate that state parity laws do not affect the likelihood of a child receiving outpatient mental health services. Among the subset of children identified with a greater need for mental health care, the effect of parity appears to be somewhat larger in magnitude but remains insignificant. DISCUSSION: State mental health parity policies aim to improve equity in private insurance coverage for mental health care and could provide a mechanism for reducing unmet need among children with mental health care disorders. Yet, our results suggest these policies do little to affect rates of use. IMPLICATIONS FOR HEALTH CARE PROVISION AND UTILIZATION: Parity policies do not appear to represent a sufficient strategy for addressing access to mental health care among children and adolescents. IMPLICATIONS FOR HEALTH POLICIES: Developing new approaches to encourage the receipt of high value mental health care in youth constitutes a persisting challenge under both public and private insurance arrangements. IMPLICATIONS FOR FURTHER RESEARCH: Since other research has shown that state parity laws are helpful in reducing the family financial burden of caring for a mentally ill child and do not drive up total health care costs, these policies serve a vital function. However, given persistent access problems in the child population, research aimed at increasing rates of mental health services use is needed. In addition, future research on parity laws should consider whether, conditional on having a visit, children in parity states are likely to have more visits compared with other children. PMID- 18509214 TI - The effect of a three-tier formulary on antidepressant utilization and expenditures. AB - BACKGROUND: Health plans in the United States are struggling to contain rapid growth in their spending on medications. They have responded by implementing multi-tiered formularies, which label certain brand medications 'non-preferred' and require higher patient copayments for those medications. This multi-tier policy relies on patients' willingness to switch medications in response to copayment differentials. The antidepressant class has certain characteristics that may pose problems for implementation of three-tier formularies, such as differences in which medication works for which patient, and high rates of medication discontinuation. AIMS OF THE STUDY: To measure the effect of a three tier formulary on antidepressant utilization and spending, including decomposing spending allocations between patient and plan. METHODS: We use claims and eligibility files for a large, mature nonprofit managed care organization that started introducing its three-tier formulary on January 1, 2000, with a staggered implementation across employer groups. The sample includes 109,686 individuals who were continuously enrolled members during the study period. We use a pretest posttest quasi-experimental design that includes a comparison group, comprising members whose employer had not adopted three-tier as of March 1, 2000. This permits some control for potentially confounding changes that could have coincided with three-tier implementation. RESULTS: For the antidepressants that became nonpreferred, prescriptions per enrollee decreased 11% in the three-tier group and increased 5% in the comparison group. The own-copay elasticity of demand for nonpreferred drugs can be approximated as -0.11. Difference-in differences regression finds that the three-tier formulary slowed the growth in the probability of using antidepressants in the post-period, which was 0.3 percentage points lower than it would have been without three-tier. The three tier formulary also increased out-of-pocket payments while reducing plan payments and total spending. DISCUSSION: The results indicate that the plan enrollees were somewhat responsive to the changed incentives, shifting away from the drugs that became nonpreferred. However, the intervention also resulted in cost-shifting from plan to enrollees, indicating some price-inelasticity. The reduction in the proportion of enrollees filling any prescriptions contrasts with results of prior studies for non-psychotropic drug classes. Limitations include the possibility of confounding changes coinciding with three-tier implementation (if they affected the two groups differentially); restriction to continuous enrollees; and lack of data on rebates the plan paid to drug manufacturers. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: The results of this study suggest that the impact of the three tier formulary approach may be somewhat different for antidepressants than for some other classes. IMPLICATIONS FOR HEALTH POLICY: Policymakers should monitor the effects of three-tier programs on utilization in psychotropic medication classes. IMPLICATIONS FOR FURTHER RESEARCH: Future studies should seek to understand the reasons for patients' limited response to the change in incentives, perhaps using physician and/or patient surveys. Studies should also examine the effects of three-tier programs on patient adherence, quality of care, and clinical and economic outcomes. PMID- 18509215 TI - Methodological advances in unit cost calculation of psychiatric residential care in Spain. AB - BACKGROUND: The care of the severe mentally ill who need intensive support for their daily living (dependent persons), accounts for an increasingly large proportion of public expenditure in many European countries. AIMS OF THE STUDY: The main aim of this study was the design and implementation of solid methodology to calculate unit costs of different types of care. To date, methodologies used in Spain have produced inaccurate figures, suggesting few variations in patient consumption of the same service. METHODS: An adaptation of the Activity-Based Costing methodology was applied in Navarre, a region in the North of Spain, as a pilot project for the public mental health services. A unit cost per care process was obtained for all levels of care considered in each service during 2005. The European Service Mapping Schedule (ESMS) codes were used to classify the services for later comparisons. Finally, in order to avoid problems of asymmetric cost distribution, a simple Bayesian model was used. RESULTS: As an illustration, we report the results obtained for long-term residential care and note that there are important variations between unit costs when considering different levels of care. Considering three levels of care (Level 1-low, Level 2-medium and Level 3 intensive), the cost per bed in Level 3 was 10% higher than that of Level 2. DISCUSSION: The results obtained using the cost methodology described provide more useful information than those using conventional methods, although its implementation requires much time to compile the necessary information during the initial stages and the collaboration of staff and managers working in the services. However, in some services, if no important variations exist in patient care, another method would be advisable, although our system provides very useful information about patterns of care from a clinical point of view. CONCLUSIONS: Detailed work is required at the beginning of the implementation in order to avoid the calculation of distorted figures and to improve the levels of decision making within the Health Care Service. IMPLICATIONS FOR HEALTH CARE POLICY AND FORMULATIONS: As other European countries, Spain has adopted a new care system for the dependent population. To finance this new system, reliable figures must be calculated for each type of user in order to establish tariffs or public prices. This study provides a useful management tool to assist in decision making. IMPLICATIONS FOR FUTURE RESEARCH: The methodology should be implemented in other regions of Spain and even in other countries in order to compare our results and validate the cost system designed. PMID- 18509216 TI - Effectiveness and cost of atypical versus typical antipsychotic treatment for schizophrenia in routine care. AB - BACKGROUND: In two recent randomised clinical trials, a meta-analysis and in an effectiveness study analysing routine data from the U.S. Veterans Administration the superiority of the newer atypical drugs over typical antipsychotic drugs, concerning both their efficacy and their side-effect profile, has been questioned. AIMS OF THE STUDY: To analyse the effectiveness and cost of atypical versus typical antipsychotic treatment for schizophrenia in routine care. METHODS: Cohort study using routine care data from a statutory sickness fund with 5.4 million insured in Germany. To be included, patients had to be discharged with a diagnosis of schizophrenia in 2003 and fulfil membership criteria. Main outcome measures were rehospitalisation rates, mean hospital bed days, mean length of stay, cost of inpatient and pharmaceutical care to the sickness fund during follow-up and medication used to treat side-effects. RESULTS: 3121 patients were included into the study. There were no statistically significant differences in the effectiveness of atypical and typical antipsychotics on rehospitalisation during follow-up (rehospitalisation rate ratio 1.07, 95% confidence interval 0.86 to 1.33). However, there were consistent observations of atypical antipsychotics being more effective for severe cases of schizophrenia (14.6% of study population; >61 prior bed days per year in 2000-2002) in the follow-up period, whereas for the other severity strata typical antipsychotics seemed more effective in reducing various rehospitalisation outcomes. Patients treated with atypical antipsychotics received significantly less prescriptions for anticholinergics or tiaprid (relative risk 0.26, 95% confidence interval 0.18 to 0.38). DISCUSSION: The effectiveness of atypical antipsychotics for schizophrenia on rehospitalisation measures appeared similar to that of typical antipsychotics. With the exception of severe cases, the higher costs for atypical antipsychotics were not offset by savings from reduced inpatient care. Major limitations include the lack of statistical power for subgroup analyses, the lack of clinical severity scale data and of life-course medical history data which both increase the risk of residual confounding by disease severity. CONCLUSIONS: This study provides evidence that the effectiveness of atypical and typical antipsychotics measured in terms of hospital readmissions appears to be similar in routine care. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: From a clinical perspective, this study provides evidence that the effectiveness of atypical and typical antipsychotics measured in terms of hospital readmissions appears to be similar in routine care. IMPLICATIONS FOR HEALTH POLICIES: Routine data studies can yield valuable information for policy decision-makers on the costs and the effectiveness of pharmaceuticals in routine care, complementing efficacy data from randomised clinical trials currently used for licensing and reimbursement decisions. IMPLICATIONS FOR FURTHER RESEARCH: The non-significant differences in the effectiveness of atypical compared to typical antipsychotics according to severity of disease should be investigated in a prospective observational study or in a randomised clinical trial. PMID- 18509217 TI - [Adaptive Wiener filter based on Gaussian mixture distribution model for denoising chest X-ray CT image]. AB - In recent decades, X-ray CT imaging has become more important as a result of its high-resolution performance. However, it is well known that the X-ray dose is insufficient in the techniques that use low-dose imaging in health screening or thin-slice imaging in work-up. Therefore, the degradation of CT images caused by the streak artifact frequently becomes problematic. In this study, we applied a Wiener filter (WF) using the universal Gaussian mixture distribution model (UNI GMM) as a statistical model to remove streak artifact. In designing the WF, it is necessary to estimate the statistical model and the precise co-variances of the original image. In the proposed method, we obtained a variety of chest X-ray CT images using a phantom simulating a chest organ, and we estimated the statistical information using the images for training. The results of simulation showed that it is possible to fit the UNI-GMM to the chest X-ray CT images and reduce the specific noise. PMID- 18509218 TI - [Elimination of bowel fluid artifact on abdominal DWI using oral contrast agent ferric ammonium citrate]. AB - The clinical usefulness, as well as the advantages and limitations of DWI in the abdomen and pelvis, currently are under discussion. One of the limitations is the artifact from bowel fluid that may obscure lesion signals or make it difficult to detect them. The purpose of this study was to examine evidence as to whether an oral contrast agent of ferric ammonium citrate could eliminate this artifact from the bowel loop. The study consisted of a phantom study and clinical study. The density of the phantom compounded it to 1-6 times, 8 times and 12 times. We changed the density with ferric ammonium citrate to find the best density. The pulse sequence used SE-PEI and, variable parameter changed TE and b value. The signal intensity of the phantoms was visually analyzed. We took the results to an outside observer as a clinical study and confirmed the effect visually. When a signal of ferric ammonium citrate extended TE time and increased b value, it fell. As it thickened the density of phantom, it was the result that the signal deteriorated, and was good. However, an artifact from ferric ammonium citrate appeared when we exceeded 5x density. The signal of the bowel fluid artifact disappeared by having a normal person take the 4x density as in the clinical study. Elimination of bowel fluid artifact on abdominal DWI was able to suggest the possibility of taking the ferric ammonium citrate solution at 4x density. PMID- 18509219 TI - [Examination of R-L shunt ratio mensuration using dynamic images in lung perfusion scintigraphy]. AB - OBJECTIVE: We examined a method of quantitative analysis that used dynamic and static images to measure the Right-left (R-L) shunt ratio in lung perfusion scintigraphy. METHODS: A total of 18 patients suspected of having R-L shunt brought on by congenital heart disease were referred for lung perfusion scintigraphy from November 2005 to February 2007 at our hospital. We acquired anterior dynamic images, set ROIs in all fields of view, and measured the highest count. We acquired anterior and posterior static images of the chest and measured lung counts. We considered the highest count on dynamic images to be the total injection dose per one second, normalized the acquisition time of static images, and calculated the R-L shunt ratio. We weighed the measurement results of the R-L shunt ratio obtained by the dynamic method against that of the whole-body method by t-test and the least-squares method. RESULTS: When we used a posterior static image in the dynamic method, we found that the coefficient of correlation (r) was 0.9831. The result of the t-test was that there was no significant difference at a level of significance of 1%. DISCUSSION: We think the use of the dynamic method is possible in R-L shunt ratio measurement and can shorten the examination time. PMID- 18509220 TI - [Influence of mechanical effect due to MRI-magnet on tattoo seal and eye makeup]. AB - The purpose of our study was to assess the mechanical effect on tattoo seals and eye makeup caused by a spatial magnetic gradient in the magnetic resonance imaging (MRI) system. Seven kinds of tattoo seals and three kinds of eye makeup, i.e., mascara, eye shadow, and eyeliner were used. On a 3.0-Tesla MRI, we determined these deflection angles according to a method established by the American Society for Testing and Materials (ASTM) at the position that produced the greatest magnetically induced deflection. Eighty-five percent of the tattoo seals showed deflection angles greater than 45 degrees of the ASTM guidelines, and the mascara and eye shadow showed over 40 degrees. This was because these contained ferromagnetic pigments such as an iron oxide, but those translational forces were very small owing to slight mass. However, it is desirable that these should be removed before MRI examination to prevent secondary problems. PMID- 18509221 TI - [Joint symposium "a benefit and risk of a medical exposure"]. PMID- 18509222 TI - [Effective communication to ease patients anxiety"]. PMID- 18509223 TI - [Education of skeletal radiology (the lower limbs)]. PMID- 18509224 TI - [Radiographic examinations for oral and maxillofacial lesions--interpretation of intraoral radiographs-]. PMID- 18509226 TI - [Fundamental technology for computer-aided diagnosis. 2. Filtering in spatial domain]. PMID- 18509225 TI - [Cardiac radionuclide imaging]. PMID- 18509227 TI - [The present situation of PET examination by delivery FDG]. PMID- 18509228 TI - [Negligence in the dispatch of an ambulance]. PMID- 18509229 TI - [DICOM Standards Committee Attendance Report--how DICOM standards are developed and maintained-]. PMID- 18509230 TI - Quantifying nanomolar levels of nitrite in biological samples by HPLC-Griess method: special reference to arterio-venous difference in vivo. AB - Nitrite (NO(2)(-)) is assumed to play an important role in regulation of vascular tone as a reservoir of nitric oxide (NO). To examine its physiological contribution, however, a sensitive method is required for determination of the true level of NO(2)(-) in biological samples. To this end, practical consideration to avoid NO(2)(-) contamination through the quantification procedure is important. We present here a highly sensitive and accurate method for determining NO(2)(-) in plasma by improving the HPLC-Griess system with minimal NO(2)(-) contamination in the samples. The system achieved high sensitivity (detection limit of 2 nM and sensitivity to 1 nM) and complete separation of the NO(2)(-) signal peak by modifying the system setup and mobile phase. Using this method, we achieved acceptable quantification of low NO(2)(-) levels in plasma. Deproteinization by ultrafiltration and exposure to atmosphere before measurement were identified as the major sources of NO(2)(-) contamination during sample processing. We addressed these issues by the use of methanol for deproteinization and gas-tight caps. These countermeasures allowed us to detect small arterio-venous NO(2)(-) differences in rabbit plasma that may indicate kinetic difference of NO(2)(-) in a small number of samples (n = 6). This difference became prominent when NO(2)(-) or a NO releasing agent, NOR1, was intravenously applied. Our results indicate that application of a sensitive method with careful handling is important for accurate determination of NO(2)(-) and that our method is applicable for further examination of the kinetic features of NO(2)(-) in vivo. PMID- 18509231 TI - Electrophysiological properties of the right atrial septum in patients with atrial tachyarrhythmias. AB - Common-type atrial flutter (AFL) is a type of atrial tachyarrhythmia with counterclockwise rotation around the tricuspid annulus within the right atrium (RA). It was recently reported that the electrogram voltage reduction observed in the RA was involved in the development of AFL. However, the relationship between the low voltage areas and conduction velocity during AFL has not been fully described. In this study, patients with AFL (n = 17) and without AFL (n = 4) were examined using an electro-anatomical mapping system. The patients with AFL were divided into 2 groups; AFL group (n = 8) and coronary sinus ostium (CSO) group (n = 9). The AFL group was defined as exhibiting the maintenance of AFL and the CSO group sinus rhythm before the catheter ablation. The electrogram voltages of each area in the RA (septum, and posterior and lateral walls), conduction velocity during AFL and transverse and longitudinal conduction velocities were evaluated. In the septum, the mean electrogram voltage was significantly lower in the AFL and CSO groups than in the group without AFL. Moreover, the conduction velocity during AFL was significantly slower in the septum, and both the septal transverse and longitudinal conduction velocities were significantly slower in the AFL and CSO groups than in the group without AFL. In conclusion, these findings suggest that both the slower conduction velocities and lower voltage in the RA septum may be involved in the development of AFL. Thus, ablation of the RA septum may represent a therapeutic approach of AFL. PMID- 18509232 TI - Maternal and neonatal factors associated with the high yield of mononuclear low density/CD34+ cells from placental/umbilical cord blood. AB - Placental/umbilical cord blood (CB) contains nucleated cells and hematopoietic stem/progenitor cells (CD34(+) cells). However it is difficult to predict the number of nucleated/CD34(+) cells in each CB before cell processing. Despite many previous studies from institutes affiliated with CB banks in metropolitan areas, little information is available regarding the characteristics of CB units from other medical facilities. The purpose of the present study was to analyze the maternal/neonatal factors on the yield of cells in CB units. A total of 176 CB units were obtained from single-birth and normal vaginal deliveries. Mononuclear low-density (LD) cells were separated using Ficoll-Paque within 24 hrs after CB collection and then processed for the purification of CD34(+) cells. A multiple linear regression analysis was performed to assess the correlations between the yield of cells and maternal/neonatal factors including maternal age, gravid status, duration of labor, gestational age, neonatal height and weight, cord length, and meconium in the amniotic fluid. The total LD cells per CB unit had a weak positive correlation with the maternal age of primigravidae. The total LD cells per CB unit from the primigravidae aged > or = 25 were significantly higher than those from the primigravidae aged < or = 24. The total CD34(+) cells per CB unit from the 1-gravidae were significantly higher than those from the 2-gravidae and 3-gravidae, respectively among all donors. These results indicate that the CB units from the primigravidae aged > or = 25 are more likely to contain higher yield of LD/CD34(+) cells. PMID- 18509233 TI - Cost-effectiveness of the national screening program for hepatitis C virus in the general population and the high-risk groups. AB - In Japan, the national screening for the hepatitis C virus (HCV) has been started for both the general population and the high-risk groups. Our cost-effectiveness analysis was based on the result of the screening program including 99,001 people among the general population and 42,538 people among the high risk group from 2003 to 2006. The screening was performed using the three steps of the semi quantitative HCV antibody test, the HCV core antigen test and the HCV-PCR test. A Markov model for HCV infected patients was constructed to estimate the future clinical benefits and the lifetime cost and the cost-effectiveness analysis was performed considering the recent treatment with peginterferon plus ribavirin. In the cost-effectiveness analysis, the cohort, in which the screening was implemented (= screening strategy), was compared with the similar cohort without the screening (= no-screening strategy) in both the general population and the high-risk group, stratified by age. The infection rates of the general population and the high-risk group were 0.36% and 0.81%, respectively. The incremental cost effectiveness ratio (ICER), a measure of cost-effectiveness, of the general population and the high-risk group was calculated to be from 848 to 4,825 and- 749 to 2,297 $/life expectancy gained, respectively. The treatment effectiveness, transition probabilities and the infection rate varied in the one-way sensitivity analyses, but the superiority of the screening strategy regarding the cost effectiveness was unchanged. In conclusion, the screening strategy in both the general population and the high-risk group therefore appears to be more cost effective than a no-screening strategy. PMID- 18509234 TI - Juzen-taiho-to, an herbal medicine, activates and enhances phagocytosis in microglia/macrophages. AB - Microglia are the main resident immunocompetent and phagocytic cells in the central nervous system (CNS). Activated microglia could play phagocytic roles as well as mediate inflammatory processes in the CNS. Involvement of activated microglia in the pathogenesis has been demonstrated in several neurological diseases including Alzheimer's disease (AD). Juzen-taiho-to (JTT), a traditional herbal medicine, has been reported to have effects on activating immune responses and phagocytosis. So far, little is known about the effects of this Kampo formulation JTT on microglia and in AD. In this report, we studied the effects of JTT on the activation and phagocytic functions of mouse microglia and bone marrow derived macrophages (BMM). JTT could activate microglia, which was confirmed by the prominent morphological change and increased surface expression of an activation marker CD11b. In addition, JTT was revealed to induce microglial proliferation, and enhance microglial phagocytosis of, without eliciting an excessive production of nitric oxide. Furthermore, when mice were administrated with JTT in vivo, their BMM showed more effective phagocytosis of fibrillar Abeta(1-42). These findings implicate the therapeutic potential of JTT in AD and other neurological diseases accompanied by microglial activation. PMID- 18509235 TI - A prominent elevation of glial fibrillary acidic protein in the cerebrospinal fluid during relapse in neuromyelitis optica. AB - Neuromyelitis optica (NMO) is a neurologic disease characterized by severe optic neuritis, longitudinally extended, transverse myelitis and serum aquaporin-4 (AQP4) antibody. Our recent neuropathological study revealed the extensive loss of AQP4 and glial fibrillary acidic protein (GFAP), an astrocyte-specific protein, in NMO lesions, but not in MS lesions, suggesting that severe astrocytic damage or dysfunction may be related to the pathogenesis of NMO. Here we report a patient of NMO, in which the cerebrospinal fluid (CSF) levels of GFAP were measured both during relapse of myelitis and after high-dose intravenous methylprednisolone (HIMP). The patient was a 34-year old woman with two previous episodes of optic neuritis. She developed myelitis longitudinally extending from C3 to T12 with contrast enhancement, and was AQP4 antibody-positive. In the acute phase, the GFAP level in the cerebrospinal fluid (CSF) was prominently elevated (18,966.7 ng/ml) as compared with controls (0.6 +/- 0.33 ng/ml). However, following HIMP, the clinical and MRI findings improved, and the CSF-GFAP level was near-normal (2.1 ng/ml). The CSF of myelin basic protein was also elevated in relapse (1,016.0 pg/ml), and became lower but still remained high (158.7 pg/ml) after HIMP compared with controls (3.36 +/- 3.83 pg/ml). The prominent elevation of the CSF-GFAP level in relapse of NMO, followed by its sharp decline after therapy, suggests severe astrocytic damage with a temporal profile distinct from that of the demyelinating process in NMO. CSF-GFAP may be useful as a biomarker of NMO. PMID- 18509236 TI - Emodin promotes atherosclerotic plaque stability in fat-fed apolipoprotein E deficient mice. AB - Increasing evidence indicated that plaque stabilization is attributed to the composition of the atherosclerotic plaque, and inflammation plays an important role in the formation and progress of vulnerable atherosclerotic plaque (VAP), which is prone to rupture. Emodin, an important component of traditional Chinese herb rhubarb, has obvious anti-inflammatory effect, although its effect on atherosclerotic plaque stabilization is unknown. Apolipoprotein E (ApoE) is an important component of plasma lipoprotein with anti-atherosclerosis function, and the plaque in the aorta of ApoE-deficient mice has been demonstrated with characteristics of VAP. Therefore, this study was designed to determine whether emodin can stabilize the VAP in the ApoE-deficient mice and explain the possible mechanism. After fat-fed for 13 weeks, mice were randomized into three groups (11 animals/group) and intragastrically administrated with emodin, simvastatin or distilled water for 13 weeks, respectively. The plaque stability was evaluated by the morphology and composition of atherosclerotic plaques. Additionally, the expression of peroxisomal proliferator-activated receptor-gamma (PPAR-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), and matrix metalloproteinase 9 (MMP-9) in plaques was determined by the immunohistochemistry method. We showed that emodin could decrease the lipid core area and the ratio of lipid to collagen content in plaques. In addition, emodin significantly inhibited the expression of GM-CSF and MMP-9, whereas it induced the expression of PPAR gamma in plaques. In conclusion, these results suggest that emodin can stabilize the VAP in the aortic root of ApoE-knockout mice, which is probably due to its anti-inflammatory effect. PMID- 18509237 TI - School children's salt intake is correlated with salty taste preference assessed by their mothers. AB - Salt intake in childhood is a risk factor for developing hypertension later in life. As health education for children to decrease salt intake, it is important for them to know the relationship between salty taste preference and salt intake. The objective of this study was to investigate the relationship between children's salty taste preference and their salt intake. We employed a cross sectional study design, and the subjects were 199 elementary school children (5th or 6th grade) and their mothers. The amount of salt intake was estimated by the amount of urinary sodium excretion. Children's salty taste preference was assessed 1) by asking children about their own salty taste preference as well as measuring their threshold level of salty taste, and 2) by their mothers' observation of their salt intake behavior using a questionnaire. The salt intake and salt taste threshold of children who liked a salty taste were similar to those in children who disliked it. No association was found between the threshold level of salty taste and sodium intake. Regarding the relationship between children's salt intake and their salt intake behavior score, assessed by their mothers using a questionnaire, the high score group had a higher estimated salt intake than the low score group. In conclusion, children's salt intake may be accurately assessed by their mother's observation rather than children's own salty taste preference. This study suggests the importance of a mother's role in salt restriction education for children. PMID- 18509238 TI - Expression of MHC-I mRNA in peripheral blood lymphocytes as an early marker of acute rejection following skin transplantation in mice. AB - Early prediction of acute rejection (AR) is important in clinical practice of organ and tissue transplantation. The aims of this study were to investigate the expression of major histocompatibility complex (MHC)-I and MHC-II genes in peripheral blood lymphocytes (PBLs) following skin grafting and whether their expression can be used as early markers for AR. Skin-grafted mice were selected as an animal model and PBL samples were collected daily for up to 2 weeks post transplant. Full-thickness skin from the backs of C57BL/6 mice (H-2b) was transplanted onto that of BALB/c mice (H-2d) in allograft group (H-2b to H-2d) and in syngeneic graft group (H-2d to H-2d). The expression levels of MHC-I (H 2K, H-2D) and MHC-II (H-2Ia, H-2Ie) mRNAs were examined using real-time PCR. The histopathological changes of graft biopsies were also analyzed with hematoxylin eosin staining. The real-time PCR analysis showed that MHC-I and MHC-II mRNA levels were increased in a bimodal distribution pattern during AR in allograft group, whereas no significant changes were detected in syngeneic graft group. The level of H-2K mRNA was significantly increased at day 5 post-transplants compared with those pre-transplant controls (p < 0.01). This increase was detected 5-6 days earlier before graft rejection observed macroscopically. H-2K mRNA level was increased significantly in 93.8% of mice (61/65) in allograft group. These results indicate that the expression of MHC-I and MHC-II mRNAs is up-regulated in PBLs during AR. Especially, the expression of H-2K mRNA can be used as an early marker for AR. PMID- 18509239 TI - Portal vein hemodynamics in patients with non-alcoholic fatty liver disease. AB - Paralleling the rise in the incidence of obesity and diabetes worldwide, nonalcoholic fatty liver disease (NAFLD) is being increasingly recognized as one of the major causes of chronic liver disease. Doppler sonography is used as a diagnostic method in the non-invasive assessment of the hemodynamics of hepatic vascular flow in liver diseases. We investigated the effects of fatty infiltration in the liver on the Doppler flow hemodynamics of the portal vein. Doppler sonography of the liver and portal vein was performed in 60 subjects with NAFLD and 20 healthy volunteers (control). The patients were grouped into mild (grade 1), moderate (grade 2), and severe (grade 3) according to sonographic appearance of hepatosteatosis (n = 20 for each group). The vein pulsatility index (VPI), mean flow velocity (MFV), peak maximum velocity (V(max)), and peak minimum velocity (V(min)) of the portal vein were significantly lower in patients with NAFLD than those of the controls (p < 0.001). The VPI was 0.20 in the patients and 0.31 in the control. The MFV was 12.3 cm/sec in the patients and 16.5 cm/sec in the control group. The portal vein flow was found to be decreased as the grade of fatty infiltration increased for VPI (r = -0.946, p < 0.001), MFV (r = -0.951, p < 0.001). The alteration in Doppler waveform pattern of portal vein with fatty liver population suggests reduced vascular compliance in the liver. PMID- 18509240 TI - Application of fluorescence polarization immunoassay for determination of methotrexate-polyglutamates in rheumatoid arthritis patients. AB - Rheumatoid arthritis (RA) is a chronic disease characterized by the painful joints, inflammation, uncontrolled proliferation of synovial tissue and multisystem comorbidities. Weekly low-dose methotrexate (MTX) has been established as effective treatment in RA patients. MTX is converted to gamma glutamyl polyglutamates, an active form of MTX, through the action of folylpolyglutamate synthetase in the cells. MTX-polyglutamates (MTX-PGs) in red blood cells (RBCs) may be useful as a therapeutic marker of RA. However, the previously reported methods for the quantification of MTX and MTX-PGs in RBCs are impractical for clinical use due to time-consuming, laborious and high cost. We attempted to apply a method with the commercially available fluorescence polarization immunoassay (FPIA) kit. We found that anti-MTX monoclonal antibody showed the reactivity to 4-amino-10-methylpteroylheptaglutamic acid (MTX-PG(7)) as equal to MTX. Good agreement was observed in the concentration-response curves between MTX and MTX-PG(7) spiked samples. Accordingly, the anti-MTX monoclonal antibody for FPIA appeared to show the equal reactivity to MTX and MTX-PGs. The recoveries of MTX and MTX-PG(7) from RBCs were 99.0% and 94.1%, respectively. Furthermore, we determined total MTX-PGs concentrations in RBCs of 71 patients with RA treated with weekly pulse MTX. Total MTX-PGs concentrations in 70% of the patients were found to be more than 50 nM that is the lower limit of MTX-PGs concentration in RBCs for expected therapeutic outcome. The routine measurement of total MTX-PGs concentration in RBCs might be useful for prediction about therapeutic outcome of MTX in RA patients. PMID- 18509241 TI - Benefit of combined cardiac rehabilitation on exercise capacity and cardiovascular parameters in patients with type 2 diabetes. AB - Favorable effects of exercise training on cardiovascular prognosis have been reported repeatedly in patients with diabetes mellitus type 2 (DM2). However, little is known about the cardiovascular rehabilitation effects in diabetic patients with coronary artery disease (CAD). This study has evaluated the benefits of combined aerobic-resistance training in two groups of patients- diabetics and non-diabetics--after percutaneous coronary intervention (PCI). Changes in exercise capacity parameters, resting cardiovascular and anthropometrical parameters were evaluated in 77 patients who completed 12-weeks of combined aerobic-resistance training: 32 patients with DM2 (DM) and 45 patients without DM2 (NDM). Significant improvements in exercise capacity (total peak workload [W(peak)], peak workload per kg of body weight [W(peak)/kg], total peak oxygen uptake [VO(2peak)], peak oxygen uptake per kg of body weight [VO(2peak)/kg]) were found in both DM and NDM (p < 0.01 and p < 0.001, respectively). The decrease in resting heart rate (HR(rest)), resting systolic (SBP(rest)) resting diastolic (DBP(rest)) blood pressures, body weight (BW) and BMI in the DM group was not statistically significant. However, there was a statistically significant decrease in SBP(rest), BW and BMI in the NDM group. In conclusion, this study demonstrated similar beneficial effects of combined cardiovascular training on exercise capacity in patients with or without type 2 diabetes mellitus. Our results suggest that the combined cardiac training is well tolerated and useful in secondary prevention in patients with DM2 and CAD. PMID- 18509242 TI - Hyperbaric oxygen therapy decreases QT dispersion in diabetic patients. AB - Diabetes mellitus is frequently associated with the malignant ventricular arrhythmias and sudden death. The QT dispersion is the difference between the longest and shortest QT interval calculated from the standard 12-lead electrocardiogram. The QT dispersion is suggested as an index of myocardial electrical activity. An increase in QT dispersion is associated with the malignant ventricular arrhythmias and sudden cardiac death. Diabetic patients receive hyperbaric oxygen (HBO) therapy for non-healing lower extremity ulcers. The aim of this study was to determine the effect of HBO therapy on QT dispersion in diabetic patients. Thirty diabetic patients (18 male and 12 female, 59.9 +/- 10 years), who were planning to undergo ten sessions of HBO therapy in two weeks for non-healing lower extremity ulcers, were consecutively enrolled into the study. The 12-lead resting electrocardiography recordings were taken before the first HBO therapy and after the 10th HBO-therapy session. QT intervals were measured on electrocardiogram. QT intervals were corrected for heart rate by using Bazett's formula (corrected QT [QTc] = QT/ radical R - R [seconds]). QTc dispersion was significantly decreased from 59.8 +/- 17.4 msec to 52.2 +/- 15.5 msec after ten sessions of HBO therapy (p < 0.05). However, maximum QTc, minimum QTc and mean QTc did not change significantly after HBO therapy. We have concluded that HBO therapy may reduce the risk of malignant ventricular arrhythmia and sudden cardiac death in diabetic patients when applied repetitively. PMID- 18509243 TI - Neuroglobin in the rat brain (II): co-localisation with neurotransmitters. AB - In an accompanying article, we found that neuroglobin (Ngb) was expressed in a few well-defined nuclei in the rat brain. Here, we show by use of immunohistochemistry and in situ hybridisation (ISH) that Ngb co-localise with several specific neurotransmitters. Ngb co-localise consistently with tyrosine hydroxylase (TH) in the noradrenergic/adrenergic A1/C1 and A2/C2; the noradrenergic A5, A6 and A7. Ngb were not observed to co-localise TH in the dopaminergic A8-A16 cell populations. Ngb were only seen to co-localise with choline acetyltransferase (ChAT) in the laterodorsal tegmental nucleus (LDTg) and in the pontine tegmental nucleus (PPTg). Many Ngb-ir neurones co-localised with neuronal nitric oxide synthase (nNOS) in the LDTg, whereas fewer Ngb-ir neurones co-localise nNOS in the anterior basomedial (BMA) and the posterodorsal medial (MePD) amygdaloid nucleus, in the medial preoptic area (MPA) and in part of the lateral hypothalamus (LH). Ngb-ir neurones co-localise heme oxygenase 1 (HO-1) in the LDTg and locus coeruleus. Ngb-ir neurones co-localise hypocretin-1 (Hcrt1) in the perifornical (PeF) and perifornical lateral hypothalamus (PeFLH). Within the LH, Ngb-ir neurones co-localised melanin concentration hormone (MCH). A few Ngb ir perikarya in the paraventricular hypothalamic nucleus (PVN) co-localised arginine vasopressin (aVP). Ngb were not observed to co-localise with serotonin, vasointestinal peptide (VIP), or cocaine amphetamine-regulated transcript (CART) at any places. In the present study, we found no evidence that one or more particular neurotransmitters are coupled 100% to Ngb or that Ngb is coupled 100% to a specific neurotransmitter. Based on these findings, we suggest that Ngb could be involved in some sort of regulation of the sleep-wake cycle. Secondly, that Ngb in some neurones is involved in regulation of gaseous neurotransmission, and that this in any given case only involves a subset of neurones. To us this indicates that the cellular and physiological function of Ngb in different subsets of neurones might not be identical, or that all neurones containing Ngb has one thing in common that we at presently not are aware of. PMID- 18509244 TI - Coenzyme Q10 administration suppresses both oxidative and antioxidative markers in hemodialysis patients. AB - BACKGROUND: Increased oxidative stress is associated with various complications in hemodialysis (HD) patients. METHODS: We examined the effect of coenzyme Q10 (CoQ10) administration on the plasma oxidative products and antioxidant capacity in 36 HD patients for 6 months. RESULTS: The advanced oxidation protein products (AOPP), malondialdehyde and the percentage of ubiquinone in total CoQ10 were significantly higher in HD patients than in healthy subjects before administration (0 month). Oxygen radical absorbing capacity (ORAC) and Trolox equivalent antioxidant capacity (TEAC), indicators of total antioxidant capacity, were also paradoxically higher in the HD patients at 0 month. AOPP and the percentage of ubiquinone significantly decreased during CoQ10 administration, but increased again after the discontinuation. ORAC and TEAC were also decreased during CoQ10 administration. CONCLUSION: The CoQ10 administration was partially effective for suppressing the oxidative stress in HD patients. The unexpected decrease of ORAC and TEAC by CoQ10 seemed to be associated with a decreased oxidative stress. PMID- 18509245 TI - Intravascular histiocytosis associated with rheumatoid arthritis. PMID- 18509246 TI - The endoplasmic reticulum stress-mediated apoptosis signal pathway is involved in sepsis-induced abnormal lymphocyte apoptosis. AB - BACKGROUND/AIM: The mechanisms of abnormal lymphocyte apoptosis in sepsis are only partially defined. The present study was designed to investigate whether the endoplasmic reticulum (ER) is implicated in the extensive apoptosis of lymphocytes in sepsis. METHODS: C57BL/6 mice were randomized into cecal ligation and puncture (CLP) and sham operation groups. Apoptosis was detected by the TUNEL method and flow cytometry. The expression of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was detected by RT-PCR and Western blot. The splicing of X box-binding protein-1 (XBP1) mRNA was detected by RT-PCR. RESULTS: A high degree of lymphocyte apoptosis was observed in the CLP group. Marked induction of GRP78 and accumulation of spliced XBP1 mRNA were observed in the splenocytes from septic mice, indicating activation of unfolded protein responses. Furthermore, both CHOP and its mRNA were markedly upregulated in the CLP group, suggesting that the ER stress response switched to a proapoptotic response. CONCLUSION: These data demonstrate activation of the unfolded protein response in lymphocytes and that ER stress may contribute to abnormal lymphocyte apoptosis during sepsis. Accordingly, the ER stress-mediated apoptosis pathway may be a novel target in clinical prevention and therapy of sepsis-induced lymphocyte apoptosis. PMID- 18509247 TI - Hospital deaths of people aged 90 and over: end-of-life palliative care management. AB - BACKGROUND: In developed countries, hospital deaths at very advanced age are increasingly common. Few studies have addressed end-of-life care in very elderly patients with non-cancer chronic diseases. OBJECTIVE: To evaluate the circumstances related to end-stage death of non-cancer nonagenarians in an acute care hospital. The results were compared with those from a sample of younger patients. METHODS: We conducted a prospective assessment in two teaching hospitals of the written instructions for the following actions: do not resuscitate (DNR) orders, the graduation of therapeutic decisions, information provided to relatives about prognosis, total withdrawal of normal drug therapy and provision of palliative care. RESULTS: 80 patients over 89 years of age with end-stage congestive heart failure (57.5%) or dementia (42.5%) were included. The control group comprised 52 younger patients (65-74 years). DNR orders were specified in 56% of cases, graduation of therapeutic decisions in 35%, and knowledge of relatives regarding the prognosis in 61%. Drug therapy was withdrawn in 66% of cases and terminal palliative care was initiated in 69%. In the nonagenarians who died, we detected a predominance of females (p = 0.001), a higher percentage of DNR orders (p = 0.02) and a higher percentage of graduation of therapeutic measures (p = 0.02) in comparison with younger patients. CONCLUSION: Our results indicate that there are marked differences according the palliative care provided to oldest-old patients with end-stage non-cancer chronic diseases admitted to an acute care hospital. In any case, care should be improved for both age groups. PMID- 18509248 TI - Endothelial function in patients with proteinuric primary glomerulonephritis. AB - BACKGROUND: Cardiovascular disease is the commonest cause of mortality among patients with end-stage renal disease. Endothelial function and inflammation have previously been shown to be abnormal among such individuals, and are known to be important factors in the progression of atherosclerosis. The aim of this study was to assess endothelial function early in the natural history of renal disease. METHODS: Patients with primary glomerulonephritis, and healthy controls were recruited. In addition to routine laboratory assessment of renal function and proteinuria, assays were undertaken to measure CRP, vWF, VCAM and ICAM. Furthermore, a direct assessment of microvascular endothelial function was undertaken, using laser Doppler imaging to measure perfusion to areas of skin under the influence of transdermally delivered vasodilator agents. RESULTS: Data were collected from 39 patients and 22 controls. No patient was taking anti platelet agents, statins or angiotensin-converting enzyme inhibitors at the time of endothelial function assessment. All 3 biomarkers of endothelial function were significantly elevated in the patient group compared to controls: ICAM 455 versus 359 ng/ml (p = 0.009), VCAM 1,101 versus 771 ng/ml (p = 0.007) and vWF 184 versus 125 IU/ml (p < 0.001). These differences remained significant after adjusting for blood pressure and body mass index. Endothelium-dependent and endothelium independent vascular responses were blunted in the patient group, compared to controls (AUC: 2,204 vs. 3,721 PU for dependent and 2,190 vs. 3,555 PU for independent responses). CONCLUSIONS: Microvascular endothelial and vascular smooth muscle function is abnormal in patients with primary glomerulonephritis and moderate proteinuria but well-maintained renal function. We believe these findings to be of particular importance as they compare 2 well-matched groups in the absence of the confounding influence of drugs known to affect endothelial function. PMID- 18509249 TI - Expression of the WASP verprolin-homologues (WAVE members) in human breast cancer. AB - BACKGROUND: The WASP family proteins have been indicated to play a vital role in the formation of membrane protrusions required for cell locomotion. WAVE proteins are an important subfamily that also plays a crucial role in actin polymerisation, which is vital to cell migration. However, not much is known about the clinical significance of this subfamily in cancers. We report, for the first time, the expression of the WAVE molecules, at protein and mRNA levels, in human breast cancer. MATERIALS AND METHODS: The expression of the 3 WAVE molecules at the mRNA and protein levels in a cohort of 122 human breast cancers and 32 normal breast tissues were analysed and correlated with the patients' pathological and clinical information as well as outcome (120 months follow-up). RESULTS: All 3 WAVE molecules were detected in mammary tissues. WAVE2 transcripts were expressed in high levels in all breast tumours. Over-expression of WAVE2 was seen in node-positive cases as well as in moderately and poorly differentiated tumours. Also, high levels of WAVE2 expression were associated with death due to disease (p = 0.02) at follow-up. No distinct associations were found between the WAVE1 and WAVE3 transcripts and the breast cancer cells. PMID- 18509250 TI - Pivotal role of the international pancreatologist in shaping the American landscape in our field. PMID- 18509251 TI - Pancreatic pain: a mini review. AB - Pain management is one of the corner stones in the treatment of pancreatitis. There are a variety of pharmacological and non-pharmacological strategies to manage the symptoms. Recognizing the type of pain, nociceptive or neuropathic, is essential for appropriate treatment. The pharmacological armamentarium currently available is substantial and includes adjuvant analgesics, non-steroidal anti inflammatories, and opioids that are customized to the etiology of the pain. When pain relief is suboptimal with pharmacological interventions, celiac block and other interventions should be considered. In acute pancreatitis the use of opioids is widely accepted while its use in chronic states is more controversial. When opioids are utilized, special care has to be taken for the assessment of indicators of misuse or abuse. A multidisciplinary approach to manage these complex patients will result in a high yield of success in controlling this and other symptoms. and IAP. PMID- 18509253 TI - Skin irritation and sensitization: mechanisms and new approaches for risk assessment. AB - Allergic contact dermatitis (ACD) is a common skin disease with a significant social and economic impact. In contrast to irritation, skin sensitization is a response of the adaptive immune system, in which there is a delayed T-cell mediated allergic response to chemically modified skin proteins. The chemicals that can covalently modify the skin proteins and trigger an allergic reaction are referred to as haptens or sensitizers. Attempts have been made in many countries to reduce the problems of ACD by the implementation of legislations related to skin-sensitizing chemicals, as well as by the early detection and risk assessment of substances with sensitizing properties. For many years, the simple identification of sensitizing chemicals was performed in guinea pig tests. A murine test, the local lymph node assay (LLNA), has been validated as a replacement for the guinea pig tests. Despite the recent introduction of in vitro methods for the identification of sensitizing chemicals, the LLNA results (when coupled with good exposure data) can be used as the starting point for a quantitative risk assessment. The quantitative risk assessment is aimed to identify the safe use thresholds for any potential skin sensitizer. PMID- 18509252 TI - Effects of HMG-CoA reductase inhibition by simvastatin on vascular dysfunction induced by lipopolysaccharide in rats. AB - AIMS: Statins have been identified as a potentially interesting treatment against sepsis. Here, we study the vascular reactivity of aortae from rats treated with lipopolysaccharide (LPS), 4 mg . kg(-1), following chronic administration of simvastatin (SV) 10 mg . kg(-1). METHODS: The rats were treated with either vehicle or SV for 4 weeks before administration of LPS. After 18 h, the systolic blood pressure (SBP) was measured using a tail cuff and vascular and endothelial responses of aortic rings to several agonists were studied in an organ bath. RESULTS: LPS injection decreased the SBP by 38 mm Hg and vascular response to phenylephrine (Phe) by 60%. Plasma nitrates and nitrites (NO(x)) were 3-fold higher after LPS. This attenuated response to Phe was prevented by incubation with either the inducible-nitric-oxide-synthase (iNOS)-selective inhibitor 1400W or the endothelial nitric oxide synthase (eNOS)/iNOS nonselective blocker L-NAME. The presence of endothelium did not alter these findings. Administering LPS to SV treated rats also decreased the SBP and increased the NO(x) concentration. The impaired response to Phe was restored by blocking NO synthesis in endothelium denuded but not in intact aortic rings. The response to acetylcholine demonstrated an enhanced reduction in arteries from the SV + LPS group compared with the LPS group. The inhibition of iNOS prevented acetylcholine-induced relaxation in rings from LPS-treated rats but not in those from the SV + LPS group. CONCLUSION: These results suggest that statins may reduce iNOS-mediated NO production in endothelial but not in vascular smooth-muscle cells. PMID- 18509254 TI - Fungicidal activity plus reservoir effect allow short treatment courses with terbinafine in tinea pedis. AB - Terbinafine, a synthetic allylamine, exerts fungicidal activity against dermatophytes, the causative pathogens of tinea pedis. As proven in numerous clinical trials, tinea pedis can be effectively and safely treated by topical terbinafine. In fact, a 1-week application of terbinafine 1% cream eradicated fungal pathogens at least as effectively as 4-week treatment courses with topical azole derivative antifungals and showed lower relapse rates. A new innovative single-application formulation of terbinafine 1% in a film-forming solution produces a high concentration gradient on the skin surface and enables a prolonged (up to 13 days) exposure of the skin to terbinafine. High drug penetration into the skin results in an otherwise not obtained drug reservoir in the horny layer, the location of dermatophytes in tinea pedis. Although azole antimycotics can also effectively penetrate into the horny layer of the skin, short-term therapy might not be feasible due to its primarily fungistatic activity against dermatophytes. Thus, we conclude that the high efficacy of short term treatment with terbinafine in patients with tinea pedis is possible due to its fungicidal activity coupled with a distinct reservoir formation in the upper layers of the epidermis. PMID- 18509255 TI - Claudin 2 mRNA and protein are present in human keratinocytes and may be regulated by all-trans-retinoic acid. AB - Tight junctions are composed of claudins, occludins, junctional adhesion molecules and plaque proteins. Claudin 2 protein forms a cation-selective channel which confers increased permeability in renal epithelial cells and in the intestine where its expression is restricted to the leaky epithelium. Immunohistochemical staining revealed claudin 2 staining in the granular layer of adult epidermis. Analysis of Western blots revealed bands corresponding to the molecular weight of claudin 2 in Madin-Darby canine kidney II cells, human kidney, adult skin and neonatal keratinocytes. Reverse transcriptase polymerase chain reaction on mRNA and cDNA sequence analysis found a 99% sequence homology between our cDNA and human claudin 2 (NIH BLAST sequence). Further, we show that all-trans-retinoic acid increases the expression of claudin 2 in keratinocytes in a dose-dependent manner. The discovery of claudin 2 transcript and protein in the skin could be of importance in epidermal differentiation, barrier function and pathological conditions. PMID- 18509256 TI - Characterization of rabbit ear skin as a skin model for in vitro transdermal permeation experiments: histology, lipid composition and permeability. AB - AIM: The aim of this work was to characterize rabbit ear skin in view of its use in transdermal permeation experiments. METHOD: The characterization included histological analysis of the tissue, qualitative and quantitative analysis of stratum corneum (SC) lipids, differential scanning calorimetry and permeation experiments (caffeine, nicotinamide, progesterone). As a reference, pig ear skin was used. RESULTS: The results obtained show that rabbit ear skin has a similar SC thickness compared to pig skin although the viable epidermis has a different structure. The lipid composition of rabbit SC was similar to pig SC but was characterized by a lower content of ceramides and a higher content of cholesterol esters and triglycerides. In terms of permeability, rabbit ear skin was 4-7 times less permeable to hydrophilic compounds, probably because of the higher lipophilicity of its SC. The permeability to progesterone was comparable between isolated pig epidermis and rabbit ear skin. CONCLUSION: Overall, the results obtained in this work support the usefulness of rabbit ear skin as barrier for skin penetration studies, for both lipophilic and hydrophilic permeants. PMID- 18509257 TI - Antiproliferative effect of vitamin A and D analogues on adult human keratinocytes in vitro. AB - BACKGROUND: Vitamin A and D analogues play an important role in epidermal homeostasis and are used in the treatment of various skin diseases. The failure of retinoid and vitamin D treatments is sometimes difficult to explain. METHODS: We analyzed the effect of all-trans retinoic acid (all-trans RA), 13-cis retinoic acid (13-cis RA), ergocalciferol and cholecalciferol in keratinocyte cultures established from adult donors, on the cell proliferation by means of [(3)H]thymidine incorporation and apoptosis after fluorescein diacetate/trypan blue staining. RESULTS: All tested agents exerted a dose-dependent inhibition of keratinocyte proliferation in the concentration range of 1.25-5 microM. Based on IC(50) values, the antiproliferative efficiency was as follows: cholecalciferol > ergocalciferol = all-trans RA > 13-cis RA. The observed effect of cholecalciferol and ergocalciferol, but not retinoids, involved the induction of apoptotic cell death. Combining vitamins A and D did not further increase the proliferation block and even displayed an antagonistic effect. CONCLUSION: The susceptibility of keratinocytes to the antiproliferative action of vitamins A and D was markedly different in cell cultures derived from different donors, indicating a possible predictive value of the in vitro testing for the efficiency of the clinical response to these agents. PMID- 18509258 TI - Normal intestinal epithelial cell differentiation in the absence of p21 and p27: new insights from old knock-out mice. PMID- 18509259 TI - Green tea and PUMA: a deadly combination? PMID- 18509260 TI - Inducing the uptake of the deadly executioner in cancer cells. PMID- 18509263 TI - Measles outbreak. PMID- 18509264 TI - Spinal decompression machines. PMID- 18509265 TI - Bevacizumab (Avastin) for metastic breast cancer. PMID- 18509266 TI - Sapropterin (Kuvan) for phenylketonuria. PMID- 18509267 TI - The small peptide OGP(10-14) acts through Src kinases and RhoA pathways in Mo-7e cells: morphologic and immunologic evaluation. AB - BACKGROUND: Osteogenic growth peptide (OGP) is an endogenous tetradecapeptide present in micromolar concentrations in mammalian serum; its carboxy-terminal pentapeptide, OGP(10-14), represents its physiologically active fragment. OGP(10 14) induces proliferation and differentiation in fibroblast and osteoblast cell lines, and it enhances hematopoiesis in vitro and in vivo. The signaling pathways triggered by OGP(10-14) are not yet fully known. In the present report, we evaluated the effect of OGP(10-14) on differentiation of a cancer megakaryoblast cell line and its involvement on RhoA and Src family kinases signaling pathway. MATERIAL/METHODS: Cell proliferation of the Mo-7e line was evaluated using the MTT test. Mo-7e differentiation was evaluated by microscopic observation of cell morphology and by expression of the factor VIII-related antigen. Involvement of RhoA and Src kinases on signaling pathways triggered by OGP(10-14) was analyzed using RhoA and Src family kinase (SFK) inhibitors (C3 and PP2) and an immunoperoxidase technique. RESULTS: OGP(10-14) induces expression of the factor VIII-related antigen, morphologic changes indicative of megakaryocytic differentiation, and a down-regulation of the Fyn Src kinase. These OGP(10-14) effects were prevented by C3 and enhanced by PP2. CONCLUSIONS: The anti proliferative and pro-differentiating activities of OGP(10-14) on thrombopoietin (TPO)-primed Mo-7e cells are mediated by RhoA and Src kinase pathways as demonstrated by the use of C3 and PP2. PMID- 18509268 TI - Protective effects of nebivolol on oxygen free radical-induced vasoconstrictions in vitro. AB - BACKGROUND: Disturbed ocular hemodynamics and vasospasms might be involved in the pathogenesis of glaucoma. On a clinical level there are indications for an optimization of ocular perfusion parameters in hypertensive glaucoma patients by switching a beta-adrenoceptor-antagonist therapy to nebivolol. Aim of the present study is to investigate vasoactive properties of nebivolol on ocular vasculature in vitro. Besides vasorelaxing effects, the impact of nebivolol on oxygen free radical-induced vasoconstrictions is studied. MATERIAL/METHODS: The experiments were carried out with ring preparations from porcine ciliary arteries. The preparations were placed in a myograph system and were kept under physiological conditions (pH 7.4, 37 degrees C, Krebs-Henseleit-Buffer, 1.75 mM Ca2+) and were stimulated by K+ depolarizations. The experiments were performed at a Nernst potential of -41 mV, which reflects half-maximal activation. For radical exposure, the preparations were superfused for 20 s in a specifically designed set-up with hydroxyl radicals generated by the Fenton reaction from H2O2 and Fe3+. NO synthase activity was modulated by adding L-arginine to the buffer. RESULTS: At a concentration of 10-5 M nebivolol leads to a reduction of vascular tone by -8.5+/-3.4% (n=11; P=0.016) vs. +2.6+/-1.9% (n=11; n.s.) in presence of its solvent DMSO. Nebivolol (10-5 M) reduces hydroxyl radical-induced vasoconstrictions by 53+/-10% (n=11; P<0.001). Stimulation of the NO synthase by L-arginine saturation potentiates this effect. CONCLUSIONS: Nebivolol combines vasorelaxing properties with protection against oxidative stress-induced vasoconstrictions. Both effects may be attributed to NO-releasing properties of nebivolol independently of its beta-adrenoceptor-blocking effect. PMID- 18509269 TI - The Candida albicans Ddr48 protein is essential for filamentation, stress response, and confers partial antifungal drug resistance. AB - BACKGROUND: Candida albicans is a dimorphic pathogenic fungus that causes mucosal and systemic infections. C. albicans pathogenicity is attributed to its ability to exist in different morphologic states and to respond to stress by up regulating several key genes. DDR48 is a stress-associated gene involved in DNA repair and in response to antifungal drug exposure. MATERIAL/METHODS: One allele of DDR48 was knocked out by homologous recombination that inserted a marker cassette in its position. Furthermore, reintroducing DDR48 on a plasmid created a revertant strain. Strains were grown on filamentation inducing and noninducing media, subjected to an oxidative stress challenge, injected into mice to assess virulence, and assayed for antifungal susceptibility by the E-test method. RESULTS: DDR48 was found to be haploid insufficient and possibly essential, since only a heterozygote, but not a homozygous, null mutant was generated. The mutant was filamentation defective on all hyphal media tested including serum and corn meal agar. Discrepancies in drug resistance profiles also were present: compared with the parental strain, DDR48/ddr48 heterozygote strain was susceptible in a dose-dependent manner to itraconazole and fluconazole and susceptible to ketoconazole. The mutant also appeared to be hypersensitive to a potentially lethal hydrogen peroxide challenge. However, no reduction in virulence of the mutant was observed. CONCLUSIONS: The present findings provide evidence that DDR48 is essential for filamentation, stress response, and possibly viability of C. albicans, making it a prime target for antifungal drug design. PMID- 18509270 TI - Effects on the ipsilateral testis during progression of experimental varicocele in rat. AB - BACKGROUND: We used an animal model of experimental varicocele to investigate the effects on the ipsilateral testis during the progression of experimental varicocele in rat. MATERIAL/METHODS: Experimental varicocele was induced by partial ligation of the left renal vein in rats. Rats in each control group were subjected to a sham operation. After different periods of varicocele creation (6, 12, and 18 weeks), each animal's left testis was extirpated, and Johnsen's score, ultrastructure of seminiferous tubules, intratesticular testosterone concentration, and the germ cell apoptotic index of each left testis were compared between the experimental and control groups. RESULTS: Johnsen's score and intratesticular testosterone concentrations in each experimental varicocele group were significantly lower than those in the corresponding control groups (P<0.05) and showed significant declines as the duration of varicocele gradually increased; the apoptotic index showed an opposite tendency. Ultrastructural injury of seminiferous tubules gradually became severe as the duration of varicocele gradually increased. CONCLUSIONS: Experimental varicocele caused progressive impairment of the ipsilateral testis; apoptosis mediated the pathophysiological processes of dyszoospermia. PMID- 18509271 TI - Bracing has no effect on standing balance in females with adolescent idiopathic scoliosis. AB - BACKGROUND: Since trunk morphology and curve types were reported to affect standing in adolescent idiopathic scoliosis (AIS), it is reasonable to assume that bracing could perturb standing balance in this population. The objective here was to use time- and frequency-domain analyses to test if the Boston brace affects balance in AIS. MATERIAL/METHODS: Fifteen AIS girls were fitted with a Boston brace. At the four-month follow-up, standing balance was assessed using center of pressure (COP) displacements measured from a force plate. The subjects were tested with and without the brace. RESULTS: The mean position of the COP and the sway area were similar with and without the brace. Though the first peak in the power spectra was not statistically significant, the second peak was statistically smaller with the brace in the antero-posterior direction (p=0.012) and larger along the medio-lateral axis (p=0.022). In the antero-posterior direction there was a statistically significant shift towards higher mean (p=0.012) and median (p=0.003) frequencies with the brace, whereas no difference was noted in the medio-lateral direction. Time-domain parameters did not prove useful in differentiating between the in- and out-of-brace conditions in AIS individuals. The second peak in the power spectra could be associated with an inverse double-pendulum motion. CONCLUSIONS: The time-dependent parameters used in this study did not prove useful in differentiating between in-brace and out-of brace conditions. Spectral analysis highlighted increased stiffness in the antero posterior direction and less control in the medio-lateral axis in standing balance between in-brace and out-of-brace conditions in AIS. PMID- 18509272 TI - Prognostic value of expression of p53, proliferating cell nuclear antigen, and c erbB-2 in laryngeal carcinoma. AB - BACKGROUND: The molecular mechanisms in malignant transformation of laryngeal mucosa are unknown; many clinical and pathological factors affect prognosis. We investigated a possible correlation between overexpression of p53, proliferating cell nuclear antigen (PCNA), and c-erbB-2, and the clinicopathologic features of laryngeal squamous cell carcinoma (SCC) and clarify their prognostic value. MATERIAL/METHODS: Thirty-three men with laryngeal SCC participated; all underwent primary surgery or surgery plus radiotherapy between 1999 and 2004 at our department. Paraffin sections of laryngeal SCC were immunohistochemically stained for p53, PCNA, and c-erbB-2. RESULTS: Overall, p53 overexpression was found in 16 patients. There was no relation between p53 immunohistochemical staining and tumor region. PCNA immunostaining was significantly stronger in supraglottic tumors compared with glottic tumors. Immunostaining of c-erbB-2 was not different in either location. There was no relation between the T stage of the tumor and p53 and c-erbB-2 immunostaining. However, there was a statistically significant positive correlation between the PCNA staining and T stage (stronger staining was present in T3 and T4 stages than in T1 and T2 stages). CONCLUSIONS: We could not find a statistically significant correlation between p53, PCNA, and c-erbB-2 and lymph node status, histologic differentiation, and survival rate. We demonstrated only a statistically significant positive correlation between PCNA staining and T stage. These data suggest that overexpression of p53, PCNA, and c-erbB-2 is not prognostic in laryngeal SCC. PMID- 18509273 TI - Evaluation of clinical outcomes and prognostic factors for synovial sarcoma arising from the extremities. AB - BACKGROUND: Synovial sarcoma is an uncommon tumor and remains a disease with poor prognosis. Although several prognostic factors have been previously reported, prognostic factors associated with synovial sarcoma are conflicting. This study was undertaken to evaluate our institutional clinical outcomes and to determine prognostic factors for synovial sarcoma. MATERIAL/METHODS: A series of 17 patients with synovial sarcoma arising from the extremities surgically treated between September 1979 and April 2005 was analyzed retrospectively. Data regarding patient age, sex, primary tumor site, primary tumor size, histologic subtype, management of primary tumor (surgery, chemotherapy, radiotherapy), microscopic surgical margins, previous treatment at other hospitals for synovial sarcoma and follow-up information were recorded. Furthermore, statistical analysis of survival rates and prognostic variables were examined. RESULTS: The mean and median follow-up time were 82.4 months and 79.0 months, respectively. Seven of 17 patients (41.2%) initially underwent inappropriate unplanned surgery at other hospitals. The surgical margins of the definitive surgery at our hospital were adequate in all patients. The local recurrence and metastasis rates after definite surgery with adequate surgical margins at our hospital were 5.9% and 35.3%, respectively. The 5-year and 10-year overall survival rates were 88.2% and 79.4%, respectively. A statistically significant adverse prognostic factor was undergoing initial surgical resection at other hospitals (p=0.02). CONCLUSIONS: Initial surgical treatment with adequate surgical margins by experienced surgeons for synovial sarcomas, preferably at specialized hospitals, should be considered to increase local control and improve outcome and survival. PMID- 18509274 TI - Fixation pattern in healthy subjects during microperimetry with the scanning laser ophthalmoscope. AB - BACKGROUND: Visual field (VF) examinations provide important information about diagnosis and follow-up in many ocular and visual pathway disorders. Previous studies have shown that fixation stability can be measured very exactly around the center of the fixation point with SLO. The importance of measuring fixation during microperimetry in the absence of field defects is to learn more about the normal fixation pattern. This is of interest since changes in fixation pattern due to pathology can occur prior to detectable changes in the macula or visual pathway. MATERIAL/METHODS: Thirty-one adult subjects with healthy eyes were recruited from the staff of the Section of Ophthalmology and Vision, Department of Clinical Neuroscience, Karolinska Institutet. The fixation pattern in one randomly selected eye from each subject was investigated with the SLO using the fixation control function in the microperimetry technique. RESULTS: The results showed that the fixation pattern had a mean center of gravity located at a mean absolute distance of 0.27 degrees from the fixation point (FP) and a directional predominance of the fixation pattern was found in that the fixations were more frequently distributed vertically than horizontally. CONCLUSIONS: The computerized fixation control when performing microperimetry with the SLO provides information about the fixation pattern which cannot be obtained with standard clinical perimetry techniques. PMID- 18509275 TI - Impact of clinical, psychological, and social factors on decreased Tinetti test score in community-living elderly subjects: a prospective study with two-year follow-up. AB - BACKGROUND: Balance and gait are essential to maintain physical autonomy, particularly in elderly people. Thus the detection of risk factors of balance and gait impairment appears necessary in order to prevent falls and dependency. The objective of this study was to analyze the impact of demographic, social, clinical, psychological, and biological parameters on the decline in balance and gait assessed by the Tinetti test (TT) after a two-year follow-up. MATERIAL/METHODS: This prospective study was conducted among community-living, young elderly volunteers in the centre "Investigations Preventives et Cliniques" and "Observatoire De l'Age" (Paris, France). Three hundred and forty-four participants aged 63.5 on average were enrolled and performed the TT twice, once at inclusion and again two years later. After the two-year follow-up, two groups were constituted according to whether or not there was a decrease in the TT score: the "TT no-deterioration" group comprised subjects with a decrease of less than two points and the "TT deterioration" group comprised those with a decrease of two points or more. Selected demographic, social, clinical, psychological, and biological parameters for the two groups were then compared. RESULTS: Statistical analysis showed that female sex, advanced age, high body mass index, osteoarticular pain, and a high level of anxiety all have a negative impact on TT score. CONCLUSIONS: Knowledge of predictive factors of the onset or worsening of balance and gait disorders could allow clinicians to detect young elderly people who should benefit from a specific prevention program. PMID- 18509276 TI - Database assessment of the effectiveness of brand versus generic rosiglitazone in patients with type 2 diabetes mellitus. AB - BACKGROUND: To compare the effectiveness of brand rosiglitazone maleate (BRM) versus generic rosiglitazone HCl (GRH) in patients with type 2 diabetes mellitus, using computerized records of a healthcare organization. Retrospective, longitudinal database analysis. MATERIAL/METHODS: Comparison of HbA1C reduction in patients starting treatment with either BRM (n=740) or GRH (n=306) in the years 2004-2005. RESULTS: BRM users were older (63.5+/-11 vs. 61.7+/-10 years p<0.001) and presented more cardiovascular disorders (38% vs. 25%, p<0.001) with no differences in gender distribution, rates of hypertension or use of concomitant oral hypoglycemic drugs. Use of concomitant insulin was more frequent (17.7% vs. 6.2%, p<0.0001), rates of dispensed rosiglitazone doses >4 mg/d (65.3% vs. 48.5%, p<0.001) and treatment duration was longer (9.3+/-6.2 vs. 5.2+/-2.2 months, p<0.001) with the generic formulation. Baseline HbA1C levels were higher (9.0+/-1.5 vs. 8.6+/-1.2%, p<0.001) and the absolute decrease in HbA1C levels was greater in the GRH group (-1.2+/-1.6% vs. -0.5+/-1.7%, p<0.001). On multiple regression analysis, the decrease in HbA1C (dependent variable) was associated mainly with initial HbA1C level (partial r2=0.30). Rosiglitazone formulation (partial r2=0.02), age, treatment duration and concomitant insulin (partial r2=0.006) were all significant but minor predictors, with no effect of rosiglitazone daily dose. Mean regression-predicted decreases in HbA1C (with 95% CL) were not significantly different between the two rosiglitazone formulations: 1.6% (-4.3% to +1.1%) for GRH and -1.1% (-3.8% to +1.6%) for BRM. CONCLUSIONS: In this retrospective database analysis, we found no evidence of different effectiveness of generic vs. brand rosiglitazone in lowering HbA1C levels. PMID- 18509277 TI - Melatonin secretion and non-specific immune responses are differentially expressed in corticotropin-dependent and corticotropin-independent Cushing's syndrome. AB - BACKGROUND: This study was done to clarify the relation between melatonin secretion and the hypothalamic-pituitary-adrenal axis. MATERIAL/METHODS: In this clinical study using a follow-up approach, we investigated the circadian melatonin secretion and immune parameters of patients with corticotropin dependent (Cushing's disease) and corticotropin-independent Cushing's syndrome. Plasma hormone concentrations, interleukin 1-beta (IL-1beta) and total immunoglobulin E (IgE) were determined before surgical treatment and during remission of the syndrome 1 year later. RESULTS: Patients with Cushing's disease showed mean nocturnal and diurnal melatonin plasma values similar to those of healthy controls. Only the midday level of patients (taken at 12:00) was significantly higher in comparison to controls (35.44+/-19.5 pg/mL vs 17.14+/ 3.58 pg/mL; P<0.05). In contrast, patients with corticotropin-independent Cushing's syndrome had significantly lower mean nocturnal and significantly higher mean diurnal melatonin levels (52.8+/-17.7 pg/mL and 59.2+/-28.7 pg/mL, respectively; P<0.05 and P<0.05) as compared with corresponding values for controls (101.4+/-43.1 pg/mL and 28.9+/-11.7 pg/mL, respectively). In the last group of patients, significantly higher mean IL1beta and plasma IgE concentrations (3.30+/-1.72 pg/mL and 527.8+/-474.0 IU/mL, respectively) were observed compared with controls (1.43+/-0.95 pg/mL and 35.7+/-32.1 IU/mL, respectively) (P<0.05). Remission of the hypercortisolism in these patients resulted in restoration of circadian melatonin secretion and significant reductions in plasma IL1beta and total IgE levels. CONCLUSIONS: Our results demonstrate a specific mode of melatonin secretion and different nonspecific immune responses in the 2 investigated forms of hypercortisolism, specifically, corticotropin-dependent and corticotropin-independent Cushing's syndrome. PMID- 18509278 TI - Immunohistological detection of Parvovirus B19 capsid proteins in endomyocardial biopsies from dilated cardiomyopathy patients. AB - BACKGROUND: Parvovirus B19 (B19V) has been identified as the most common virus in dilated cardiomyopathy (DCM) patients by PCR techniques. We investigated the detectability and the expression pattern of B19V proteins by immunohistology (IH) in endomyocardial biopsies (EMBs) from DCM patients, and its association with the standard proof of B19V genomes by nested PCR (nPCR. MATERIAL/METHODS: EMBs from 30 DCM patients were analyzed by nPCR for B19V genomes, and IH of B19V VP1/VP2 expression was carried out using the antibody clone R92F6. The specificity of this antibody was investigated on 293T cells transfected with pB19-M20. Positive anti-B19V IH staining (positive in n=14/46.6%) and nPCR proof of B19V genomes (positive in n=15/50%) were significantly associated (p=0.0003). RESULTS: Based on the B19V nPCR results, the sensitivity of anti-B19V-VP1/VP2 IH was 80.0%, and the specificity was 86.0%. B19V immunostaining was observed on interstitial cells in all IH positive cases, and was noted additionally on endothelial cells in 1, and on cardiomyocytes in 4 cases. CONCLUSIONS: IH detection of B19V VP1/VP2 expression provides a high association with the nPCR proof of B19V genomes in DCM patients. IH detection of B19V proteins enables a differentiation of B19V VP1/VP2 expressing cells within the myocardium. PMID- 18509279 TI - Lipid levels among African and Middle-Eastern Bedouin populations. AB - BACKGROUND: Previous studies observed higher high-density lipoprotein (HDL) levels and lower triglycerides levels among people of African ancestry. The goal of this study was to characterize lipid levels in Bedouins of African vs. Middle Eastern ethnicity. MATERIAL/METHODS: A cross-sectional study was conducted in a Bedouin primary care clinic in southern Israel, with 4470 listed individuals over the age of 21, of whom 402 (9%) were of African origin. A stratified random sample was included in the analysis. Associations between ethnicity, age, gender and lipid levels were assessed. Multiple linear regression and logistic regression models were used for multivariate analysis. RESULTS: The study included 261 African Bedouins and 406 Middle-Eastern Bedouins. (median age: 37 years, 58.6% females). The average total cholesterol and low-density lipoprotein (LDL) levels were 10 mg/dl lower among African Bedouins as compared to Middle Eastern Bedouins (total cholesterol: 168.6 vs. 179.6 mg/dl, p<0.001; LDL: 99.5 vs. 109.0 mg/dl, respectively, p<0.001). Average triglycerides levels were 36 mg/dl lower among African Bedouins as compared to Middle-Eastern Bedouins (102.8 vs. 138.9 mg/dl, respectively, p<0.001). Average HDL levels were 3 mg/dl higher among African Bedouins as compared to Middle-Eastern Bedouins (48.3 vs. 44.6 mg/dl, respectively, p<0.001). CONCLUSIONS: A lower prevalence of dyslipidemia was found in African Bedouins, as compared with Middle-Eastern Bedouins. PMID- 18509280 TI - Identification of endogenous morphine and a mu3-like opiate alkaloid receptor in human brain tissue taken from a patient with intractable complex partial epilepsy. AB - BACKGROUND: We set out to detect whether morphine is present in tissue taken from a patient with intractable temporal lobe epilepsy and to characterize the presence and nature of mu opiate receptor subtypes in this tissue. CASE REPORT: In temporal lobe tissue, resected during anteromedial temporal lobectomy for intractable focal epilepsy, morphine was identified by quantitative radioimmunoassay (RIA) coupled to electrochemical detection via high-pressure liquid chromatography (HPLC). In addition, RNA isolated from the medial and lateral temporal lobe specimens was analyzed by conventional and real time reverse transcriptase-polymerase chain reaction (RT-PCR) for the expression of different human receptor gene transcripts. RIA revealed the presence of morphine at 3.4 nanograms per gram of tissue wet weight. Using RT-PCR and a primer specifically set for the mu3 (550 base pair fragment) and mu4 (880 base pair fragment) MOR splice variants, a mu4 splice variant was identified in both brain sections. CONCLUSIONS: This human brain tissue study of a subject with temporal lobe epilepsy documents the presence of endogenous morphine and of a mu4 splice variant. These findings may have implications for our understanding of the mechanism of temporal lobe epilepsy. PMID- 18509281 TI - Laparoscopic wedge resection of a midsized gastrointestinal stromal tumor near the esophagogastric junction. AB - BACKGROUND: Gastrointestinal stromal tumors are rare neoplasms involving the entire digestive system, in particular the stomach. Generally, laparoscopic wedge resection is the preferred means of treating anterior lesions, while the preferred means of treating tumors in the posterior wall and near the esophagogastric junction remains controversial. CASE REPORT: We report the case of a patient with a gastric stromal tumor in the posterior wall of the greater curvature approximately 3 cm from the esophagogastric junction. Under intraoperative ultrasonic guidance, a large wedge resection was done using an ultrasonic scalpel; the gastric wall was reconstructed through a long intracorporeal suture. At 6-month follow-up, the patient was free from tumor recurrence. CONCLUSIONS: If a meticulous surgical technique is followed, laparoscopic wedge resection is a safe and feasible means of treating gastric stromal tumors in the posterior wall near the esophagogastric junction. PMID- 18509282 TI - Apical ballooning syndrome: a metabolic form of cardiomyopathy? AB - Takotsubo cardiomyopathy is a transient cardiomyopathy of unknown origin due to left ventricular apical ballooning. Although first described in Japanese patients, it is now diagnosed worldwide. The syndrome mimics acute ST-elevation myocardial infarction in the absence of significant epicardial coronary artery disease. Although significant efforts have been made to understand it, the pathophysiology of takotsubo cardiomyopathy remains poorly understood. We hypothesize that takotsubo cardiomyopathy may be considered a metabolic form of cardiomyopathy characterized by dysfunctional metabolism of cardiomyocytes. Evidence to support this hypothesis is given and possible therapeutic implications are discussed. PMID- 18509283 TI - Lubricating agents differ in their protection of cultured human epithelial cells against desiccation. AB - BACKGROUND: In dry eye, as a disease of the ocular surface, the instillation of artificial tears should compensate for the deficit in wetting and protect the mucosa against drying. MATERIAL/METHODS: The desiccation protection of different pharmacological substances was tested using the conjunctival epithelial cell line Chang 1-5c-4 (series 1) and the corneal cell line 2.040 pRSV-T (series 2). On confluent cell growth the cultures were wetted for 20 min with various preservative-free preparations of artificial tears The cell cultures were exposed to a constant air flow for 0, 15, 30 and 45 minutes. Cells were incubated with the vital dye Alamar Blue and subsequently absorption of the oxidised form of the dye was measured using an ELISA-Reader. RESULTS: Cell survival rates in series 1 after 0, 15, 30, 45 min were (1.02;0.81;0.35;0.32) for Artelac EDO, (0.82;0.69; 0.63;0.54) for Vidisic EDO, (0.77;0.80;0.67;0.70) for Vidisic Fluid EDO, (0.76;0.70;0.36; 0.34) for Acuolens, (0.97;0.46;0.35;0.33) for Viscofresh, (0.88;0.85;0.37; 0.33) for Hyal Drops SDU, (0.71;0.44;0.34;0.33) for PBS and in series 2 (1.03;0.84;-0.21;-0.20) for Artelac EDO, (0.89;0.92;0.93;0.86) for Vidisic EDO, (0.96;0.88;0.85;0.85) for Vidisic Fluid EDO, (1.01;0.75;-0.02;-0.03) for Acuolens, (0.98;0.17;-0.22;-0.20) for Viscofresh, (0.97;0.83;0.03;-0.21) for Hyal Drops SDU and (0.96;0.26;-0.24;-0.21) for PBS. Vidisic Fluid EDO and Vidisic EDO showed a significantly better protective effect after a drying period of 30 and 45 min. CONCLUSIONS: The protection capability of pharmacological substances against desiccation can be studied in a standardised cell culture system of human epithelial cell lines. Whether these in vitro results are conferrable to the efficacy of artificial tear drops in vivo has to be evaluated in clinical trials. PMID- 18509284 TI - Increased intramuscular and intracapsular temperature via ThermaCare Knee Wrap application. AB - BACKGROUND: Knee pain is common among the physically active as well as the older population. Heat is often used to decrease pain in these individuals. Modalities such as hot packs and whirlpools increase tissue temperature, yet are not portable. The portable ThermaCare HeatWrap for the knee has recently been marketed. We wondered if it actually heated the knee joint. MATERIAL/METHODS: A 2 x 5 factorial design was used to compare temperature changes in a treatment group (heat wrap) against a control group (no wrap) for two hours. Eleven volunteers (M=3, F=8; age=22.71+/-3.80) served in both groups. An Isothermex computer recorded temperatures to the nearest +/-0.1 degrees C every 30 seconds. Subjects rated heat perception using a 10 cm visual analog scale (VAS). To measure muscle temperature one thermocouple monofilament was inserted 1.5 cm into the vastus medialis oblique (VMO). To measure knee intracapsular temperature (KJC), a thermocouple was inserted 5 cm deep in the suprapatellar pouch. RESULTS: The mean temperature increases for the ThermaCare Knee Wrap were: VMO: 3.19+/-1.27 degrees C; KJC: 2.62+/-0.93 degrees C; Surface: 7.48+/-1.35 degrees C. The mean temperature increases for the control were: VMO: -1.99+/-1.01 degrees C; KJC: 2.10+/-0.56 degrees C; Surface: -1.15+/-0.61 degrees C. According to the VAS data, the longer the subjects wore the wrap, the warmer it felt. CONCLUSIONS: The ThermaCare Knee Wrap causes significant temperature increases in the VMO, the KJC and the skin. This portable heat product is beneficial for conditions in which intramuscular and intracapsular heating is indicated, such as pain relief. PMID- 18509285 TI - Schizophrenia and viral infection during neurodevelopment: a pathogenesis model? AB - The neurodevelopmental hypothesis of schizophrenia proposes that a portion of schizophrenia is the result of an early brain insult which affects brain development and in which several types of virus might play an etiological role. The main arguments in favor of the neurodevelopmental hypothesis and the involvement of prenatal exposure to virus infection as a risk factor for adult schizophrenia are reviewed. Schizophrenia is associated with an increased incidence of craniofacial asymmetries and dermatoglyphic irregularities which might reflect an abnormal development of the ectoderm and the neural crest as a result of a viral infection between the first and second trimester of pregnancy. The brain histology of deceased schizophrenic patients shows disturbed neuronal migration and formations such as disorganized lamina strata or ectopic pyramidal cells, abnormal expression of the neural cell adhesion molecule, and absence of gliosis. The main epidemiological arguments are derived from studies of obstetrical complications and influenza virus infection during pregnancy, both considered to be early risk factors of schizophrenia. Because no virus has been consistently linked with the pathogenesis of schizophrenia, the most plausible hypothesis is that an endemic virus could initiate schizophrenia by direct brain lesion or by triggering an autoimmune response during the neurodevelopmental period on a genetically susceptible brain. In a neurodevelopmental model, the viral hypothesis is a step toward the goal of building a comprehensive theory that integrates the environmental, genetic, immune, and neuropsychological features of schizophrenia. PMID- 18509286 TI - TCF2 gene mutation leads to nephro-urological defects of unequal severity: an open question. AB - There are several genes known to be involved in simple renal or combined renal extrarenal aberrations. Of these, the transcription factor 2 gene is expressed longer, from very early embryogenesis and throughout organ development during pregnancy. Transcription factor 2 gene encodes the hepatocyte nuclear factor-1 beta transcript, which is a member of the homeodomain-containing superfamily of transcription factors. Transcription factor 2 gene mutations may be associated with a wide variability in severity and pattern of clinical symptoms. Transcription factor 2 gene mutation may be responsible for approximately one third of children having isolated renal cysts, multicystic dysplastic kidneys, oligomeganephronia, hypo-dysplastic kidneys, horseshoe kidneys, and hyperechogenic kidneys. The wide clinical presentation of hepatocyte nuclear factor-1 beta mutations suggests a broad role of this transcription factor throughout development. The complexity of phenotypes is quite interesting because it could depend on the vast expression time of this gene derangement during fetal development or on different gene-gene and gene-environmental interactions during different stages of embryogenesis. The current literature is reviewed concerning the malformations that have been associated with transcription factor 2 gene mutations involving primarily the kidneys and occurring both in an isolated form and in association with other defective organs to characterize the patterns of this genetic disease. PMID- 18509287 TI - Clinical and pharmacological management of a high risk diabetic population undergoing percutaneous coronary interventions. AB - AIM: An optimal clinical management and monitoring of patients with diabetes mellitus (DM) and coronary artery disease (CAD) has proved to reduce the incidence of cardiovascular events. The main aim of this report is to verify the adherence to the latest guidelines recommendations in a considerable group of DM patients undergoing a percutaneous coronary intervention, stratifying the population by the degree of the DM management. METHODS: The patients recruited were those consecutive, unselected patients with type 2 DM undergoing a balloon angioplasty followed by a stent implantation, between July 2002 and December 2004. Follow-up was completed in 220 patients at 39+/-8.7 months. The follow-up interview concerned sociodemographic details, medical history, pharmaceutical treatment, major adverse cardiac and cerebrovascular events and clinical variables regarding the quality of DM management and monitoring. RESULTS: Thirty six percent of these high short and long-term risk patients received insulin treatment at index hospitalization despite a poor glucometabolic control (59% presented HbA1c >7%, mean 8+/-1.26%). The clinical management of the 204 (80%) survivors over time proved overall unsatisfactory: HbA1c, arterial blood pressure and body mass index uncommonly achieved the recommended targets (41%, 36% and 22% of the patients respectively). The DM monitoring revealed satisfactory, with the exception of the diabetic foot monitoring (performed in 38% of the patients). Concerning guideline adherence 26% of the patients proved to adhere strictly, while 38% and 36% were mildly and severely distant from these requirements. CONCLUSION: Quality of DM management absolutely necessitates to be improved to attempt reducing the high cardiovascular event rates. PMID- 18509288 TI - Prediction of functional capacity by low-dose dobutamine stress echocardiography in chronic heart failure. AB - AIM: The aim of this study was to determine if low-dose dobutamine stress echocardiography (LD-DSE) is associated with functional capacity in unselected elderly patients with chronic heart failure. METHODS: This was a prospective trial. Thirty five consecutive patients were included, with age >65 years and left ventricular dysfunction (12 ischemic), by blindly assessed LD-DSE and cardiopulmonary exercise testing (CPT). Contractile reserve was defined as a change (D) in wall motion score index (WMSI) = or <0.2 at peak dose dobutamine. At CPT treadmill exercise time, peak VO2, %Vo2 and VE/VCO2 slope were determined. Preserved functional capacity was defined as percent of maximal predicted O2 consumption (%VO2) >80%. Baseline NT-proBNP plasma levels were assessed. RESULTS: CPT variables were not related to clinical and baseline echocardiography characteristics but were related to DWMSI (exercise time, P=0.004; peak VO2, P=0.008; %VO2, P<0.001; VE/Vco2, P<0.001). Contractile reserve was present in 16 of 17 patients with preserved functional capacity (sensitivity=94%) and in 2 of 13 patients without (specificity=85%). Baseline NT-proBNP levels were lower in patients with contractile reserve (476+/-365 pg/mL) than in those without (1 345+/-1 219 pg/mL) (P=0.019), but were mildly related to CPT variables (P=0.049 and 0.027 with exercise time and %VO2, respectively). CONCLUSION: Contractile reserve elicited at LD-DSE is associated with functional capacity in unselected elderly patients with chronic heart failure. PMID- 18509289 TI - Nocturnal oxygen desaturation correlates with glycemia levels in patients with coronary artery disease. AB - AIM: Sleep-disordered breathing (SDB) has a high occurrence in patients with coronary artery disease. The aim of the present study was to examine the relationships between the overnight oxygen desaturation index (ODI) and metabolic and anthropometric parameters in a sample of patients who underwent percutaneous transluminal coronary angioplasty (PTCA). METHODS: The overnight pulse-oximetry recording ODI (desaturation >4%, length >10 s/events per hour) was performed in 29 patients with coronary heart disease, after they underwent PTCA. All study participants performed dynamic spirometric test, echocardiography, anthropometric measurements, bio impedance analysis. Fasting blood sample was collected to measure metabolic parameters. Using ODI category, the patients were grouped as follows: group N, N.=9 (ODI= or <5 events/hour); group A, N.=14 (515 events/hour). RESULTS: Among the studied patients, 69% had a nocturnal ODI>5 and 48% had a nocturnal ODI between 5 and 15 events/hour. ODI values were correlated with body mass index (BMI) (P<0.05) but not with waist to hip ratio, or percent body fat. After adjustment for BMI values, there was a significant positive correlation of ODI values with fasting glucose serum levels (P<0.05). CONCLUSION: These data suggest that early metabolic dysfunction may occur with SDB before overt clinical manifestation of underlying disease in patients with preobesity and ischemic heart disease. Patients with ischemic heart disease should be screened for SDB by ODI, also according to fasting glucose levels. PMID- 18509290 TI - Predictors of restenosis after coronary artery stenting. AB - AIM: The aim of this study was to analyze restenosis after percutaneous coronary intervention, factors related to restenosis after coronary artery stenting and the degree of the risk of restenosis were evaluated. METHODS: The study enrolled 181 patients (249 lesions) who underwent the first coronary artery stenting. Multivariate analysis was performed, and the restenotic index (RI) was calculated by combining the extracted predictors. RESULTS: Among the 181 patients (249 lesions), restenosis occurred in 89 (111 lesions) and did not occur in 92 (138 lesions). Vascular revasculation was performed in 95 restenosed target lesions in 68 patients. The mean period of follow-up angiography after the procedures was 206 days in the restenosis group and 271 days in the non-restenosis group, i.e. significantly shorter in the restenosis group. As a result of multivariate analysis, diabetes mellitus, Cr level, amount of the contrast medium used and stent diameter were selected as significant factors that independently contributed to the restenosis after coronary artery stenting. By combining these factors, the RI was calculated by the following formula for the prediction of restenosis: RI=exp (1.088xCr+0.909xdiabetes mellitus+0.871xcontrast medium+0.591xstent diameter). CONCLUSION: The risk of restenosis after coronary artery stenting can be predicted to an extent according to the RI devised in this study. PMID- 18509291 TI - The evolution of cardiac electrophysiology in the XIX century: historical highlights. PMID- 18509292 TI - Pediatric heart failure. AB - Heart failure is a clinical syndrome which presents similarities and differences between children and adults; in pediatric age the spectrum of causes of heart failure is wide and congenital heart defects are the most common etiology. Volume and pressure overload on a ''normal myocardium'' is the classical physiological pattern while myocardial contractile dysfunction of different etiology is much less observed in the pediatric population. However there are some peculiarities in clinical presentation of heart failure in infants and small children. The medical therapy cornerstones still remain loop diuretics, angiotensin-converting enzyme inhibitors, beta-blockers and digitalis. There are also some reported experiences with new inotropics drugs in acute heart failure. In pediatric cardiology there are few prospective studies on pharmacology of heart failure and the data are often extrapolated from adult large trials. Non pharmacological treatment with autonomic implantable cardioverter defibrillators and resynchronization therapy as well as the surgically implant of ventricular assist devices are increasingly employed in children. Cardiac transplantation is currently the treatment option with good outcome and long-term survival in pediatric patients with end-stage or refractory heart failure. PMID- 18509293 TI - Strategies for the treatment of acute heart failure in children. AB - Acute heart failure is life threatening in adults and in the pediatric population. It represents the final pathway of complex physiological mechanisms that vary with age and underlying diseases. Strategies for treatment of heart failure should integrate to an concept of stepwise approach to heart failure. Evolving therapies, that are changing standard of care, are discussed. This paper gives an overview on principles for treatment of congestive heart failure, like diuretics, ss-beta-blockade, and angiotensin-converting enzyme blockers and discusses the approach to acute heart failure, including new and well known inotropic agents. Mechanical circulatory support and finally heart transplantation are focused as options in the last line. PMID- 18509294 TI - Stem cells in pediatric heart failure. AB - Pediatric heart failure could be a target for regenerative therapy. Stem cell based therapy has the potential to provide functional cardiomyocytes. Whereas adult stem cells have shown no or only minimal therapeutic benefit in adults with no evidence of transdifferentiation, embryonic stem cells can differentiate to any cell type, including cardiomyocytes. However, ethical concerns and immunological problems are associated with embryonic stem cells derived from the inner cell mass of blastocysts. Recently, somatic cells could be reprogrammed to a pluripotent state (i.e. induced pluripotent stem cells) with the help of transcription factors. This technique removes ethical and probably also immunological concerns. Nevertheless extensive experimental research will be necessary before cell replacement strategies become clinically applicable. Because the underlying pathophysiology differs significantly with age, caution is warranted extrapolating data obtained in experimental models of cardiac ischemia and clinical studies in adults to the pediatric population. Pediatric heart failure has a good prognosis if causal therapy is possible. However, some forms of congenital heart disease and especially dilated cardiomyopathy still have limited therapeutic options. Almost half of children with symptomatic cardiomyopathy receive a transplant or die within two years. The authors will review the relevant stem cell sources for cell-based treatments. And, given the differences of the underlying diseases between adult and pediatric patients with heart failure, it is contemplated which condition of pediatric patients with heart failure is most likely to benefit and which cell type would be appropriate. PMID- 18509295 TI - Heart transplantation in children. AB - Cardiac transplantation has become a standard therapeutic option for a variety of cardiac conditions in childhood in which poor cardiac output without other surgical options exists in the face of maximized medical therapy. In this review, the common diagnoses resulting in heart transplantation, risk factors and outcomes of heart transplantation in children with otherwise inoperable heart disease are discussed. PMID- 18509296 TI - Coronary artery fistula: diagnostic role of transthoracic and transesophageal echocardiography. AB - Congenital coronary artery fistulas (CAFs) are rare. Some patients develop symptoms of congestive heart failure secondary to a large left-to-right shunt or myocardial ischemia from coronary artery steal in the first few years after birth. After the second decade the frequency of symptoms and complications increase. We report a case of CAF originating from the circumflex artery and draining into the coronary sinus, associated with left main coronary aneurysm. Transtho-racic and transesophageal echocardiography approach showed the origin, course, and drainage site of the CAF. This case represents a typical sample of this rare anomaly and puts into evidence the essential role of echocardiography to define and complete the angiographic diagnosis. PMID- 18509297 TI - Iatrogenic coronary-to-right ventricle fistula: benign outcome irrespective of patency? AB - Coronary perforation is an uncommon, but potentially lethal complication of percutaneous coronary interventions. Most perforations have proved to lead to intramyocardial or intrapericardial extravasation. However, perforation may also lead to direct coronary-to-right ventricle fistula, more commonly occurring in complex lesions and/or the use of atheroablative devices. To prevent this complication careful manipulation of the guidewire across the obstruction is mandatory, together with avoidance of oversized balloons and high-pressure inflations. The factors that determine the hemodynamic significance of the fistulas include size of the communication, resistance of the recipient chamber, and potential for development of myocardial ischemia. Accordingly a broad range of signs and symptoms may be referred. Despite iatrogenic fistulas are usually benign and asymptomatic, spontaneous closures are very rare. In most cases fistulas need to be treated by percutaneous or surgical closure, being the conservative management a debated option. In isolated cases serious complications have been described resulting from volume overload and distal myocardial flow impairment. We hereby describe two cases of coronary-to-right ventricle fistula occurred during percutaneous coronary intervention, highlighting full angiographic iconography and the issues involved in the management and follow-up of this iatrogenic complication. PMID- 18509298 TI - Triphasic diastolic pattern and delayed timing of myocardial relaxation identify dynamic changes of left ventricular filling pressure. AB - The case reported concerns a woman with hypertensive emergency, showing triphasic diastolic pattern with mid-diastolic velocity both at transmitral inflow and at pulsed tissue Doppler-derived septal/lateral mitral annulus. In addition, the time from EKG R wave peak to annular early diastolic velocity (Em) onset was longer than the time occurring between R peak and transmitral E velocity. E/Em ratio was 14.6 and left atrium enlarged. After blood pressure fall and clinical stabilization, the triphasic diastolic patterns were again observed. However, the timing of early diastolic velocity was the same at mitral inflow and annular tissue Doppler. E/Em ratio and left atrial volume were reduced. The present report highlights the additional value of pulsed tissue Doppler to detect alteration of diastolic properties and dynamic changes of left ventricular filling pressure. PMID- 18509299 TI - A case of thrombus outside a ligated left atrial appendage despite prolonged oral anticoagulant therapy. PMID- 18509300 TI - Right lobar pulmonitis: a possible cause of lone atrial fibrillation. PMID- 18509301 TI - Primary amebic meningoencephalitis--Arizona, Florida, and Texas, 2007. AB - Primary amebic meningoencephalitis (PAM) is a rare but nearly always fatal disease caused by infection with Naegleria fowleri, a thermophilic, free-living ameba found in freshwater environments. Infection results from water containing N. fowleri entering the nose, followed by migration of the amebae to the brain via the olfactory nerve. In 2007, six cases of PAM in the United States were reported to CDC; all six patients died. This report summarizes the investigations of the cases, which occurred in three southern tier states (Arizona, Florida, and Texas) during June-September and presents preliminary results from a review of PAM cases during 1937-2007. Because deaths from PAM often prompt heightened concern about the disease among the public, an updated and consistent approach to N. fowleri risk reduction messages, diagnosis and treatment, case reporting, and environmental sampling is needed. PMID- 18509302 TI - Detection of West Nile virus in blood donations--Puerto Rico, 2007. AB - In the United States, West Nile virus (WNV) was first detected in humans in 1999; it subsequently spread to countries of Central and South America and the Caribbean. WNV is a mosquito-borne virus that produces potentially serious clinical disease, particularly among persons aged > or =50 years. Transmission by routes other than mosquito bites, including blood transfusion, transplacental infection, organ transplant, and possibly breast milk, also have been reported. On July 19, 2007, the American Red Cross in Puerto Rico notified the Puerto Rico Department of Health (PRDH) of three persons whose blood donations were positive for WNV by nucleic acid-amplification test (NAT) screening. These three donors had the first confirmed human WNV infections detected in Puerto Rico. In response, PRDH and CDC conducted in-depth interviews of the blood donors. This report describes these human infections and other recent surveillance for transmission of WNV in Puerto Rico. Detection of WNV infections in human blood donors indicates that heightened clinician awareness, ongoing surveillance, and educational activities are needed to monitor and assess the public health threat posed by WNV in Puerto Rico. PMID- 18509303 TI - Guiding principles for development of ACIP recommendations for vaccination during pregnancy and breastfeeding. AB - The Advisory Committee on Immunization Practices (ACIP) provides advice and guidance regarding effective control of vaccine-preventable diseases, including guidance for special populations that might warrant modification of routine recommendations. One such special population is pregnant and breastfeeding women. Formulation of recommendations for vaccination of pregnant and breastfeeding women is challenging because the available scientific evidence needed to guide decisions is limited. To promote use of a consistent process and uniform terminology, the ACIP Workgroup on Vaccines during Pregnancy and Breastfeeding was established in 2007 to develop guiding principles for drafting of ACIP recommendations for vaccination of pregnant and breastfeeding women. Workgroup members included ACIP members, members of professional medical organizations, experts in the field, and CDC consultants. PMID- 18509307 TI - Molecular screening for new fusion genes in cancer. PMID- 18509304 TI - Prevention of pertussis, tetanus, and diphtheria among pregnant and postpartum women and their infants recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - In 2005, two tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccines were licensed and recommended for use in adults and adolescents in the United States: ADACEL (sanofi pasteur, Swiftwater, Pennsylvania), which is licensed for use in persons aged 11--64 years, and BOOSTRIX (GlaxoSmithKline Biologicals, Rixensart, Belgium), which is licensed for use in persons aged 10-18 years. Both Tdap vaccines are licensed for single-dose use to add protection against pertussis and to replace the next dose of tetanus and diphtheria toxoids vaccine (Td). Available evidence does not address the safety of Tdap for pregnant women, their fetuses, or pregnancy outcomes sufficiently. Available data also do not indicate whether Tdap-induced transplacental maternal antibodies provide early protection against pertussis to infants or interfere with an infant's immune responses to routinely administered pediatric vaccines. Until additional information is available, CDC's Advisory Committee on Immunization Practices recommends that pregnant women who were not vaccinated previously with Tdap: 1) receive Tdap in the immediate postpartum period before discharge from hospital or birthing center, 2) may receive Tdap at an interval as short as 2 years since the most recent Td vaccine, 3) receive Td during pregnancy for tetanus and diphtheria protection when indicated, or 4) defer the Td vaccine indicated during pregnancy to substitute Tdap vaccine in the immediate postpartum period if the woman is likely to have sufficient protection against tetanus and diphtheria. Although pregnancy is not a contraindication for receiving Tdap vaccine, health-care providers should weigh the theoretical risks and benefits before choosing to administer Tdap vaccine to a pregnant woman. This report 1) describes the clinical features of pertussis, tetanus, and diphtheria among pregnant and postpartum women and their infants, 2) reviews available evidence of pertussis vaccination during pregnancy as a strategy to prevent infant pertussis, 3) summarizes Tdap vaccination policy in the United States, and 4) presents recommendations for use of Td and Tdap vaccines among pregnant and postpartum women. PMID- 18509308 TI - New susceptibility genes for ulcerative colitis. PMID- 18509309 TI - Prader-Willi and snoRNAs. PMID- 18509310 TI - The not-so-silent X. PMID- 18509311 TI - The Hippocratic finger points the blame at PGE2. PMID- 18509313 TI - Common and rare variants in multifactorial susceptibility to common diseases. AB - Here, we give a historical overview of the search for genetic variants that influence the susceptibility of an individual to a chronic disease, from RA Fisher's seminal work to the current excitement of whole-genome association studies (WGAS). We then discuss the concepts behind the identification of common variants as disease causal factors and contrast them to the basic ideas that underlie the rare variant hypothesis. The identification of rare variants involves the careful selection of candidate genes to examine, the availability of highly efficient resequencing techniques and the appropriate assessment of the functional consequences of the implicated variant. We believe that this strategy can be successfully applied at present in order to unravel the contribution of rare variants to the multifactorial inheritance of common diseases, which could lead to the implementation of much needed preventative screening schemes. PMID- 18509314 TI - Acute coronary syndrome in ESRD patients. PMID- 18509315 TI - Autophagy is cytoprotective during cisplatin injury of renal proximal tubular cells. AB - Autophagy is a cellular process of bulk degradation of damaged organelles, protein aggregates and other macromolecules in the cytoplasm. It is thought to be a general response to stress contributing to cell death; alternatively it might act as a cytoprotective mechanism. Here we found that administration of cisplatin induced the formation of autophagic vesicles and autophagosomes in mouse kidneys. In cultured proximal tubular cells, the nephrotoxin caused autophagy in a dose- and time-dependent manner prior to apoptosis. Notably, autophagy occurred within hours of cisplatin administration but this was partially suppressed by the p53 inhibitor pifithrin-alpha, suggesting that p53 is involved in autophagic signaling. This cisplatin-induced autophagy was attenuated in renal cells stably transfected with Bcl-2, suggesting an anti-autophagic role for this well-known anti-apoptotic protein. Blockade of autophagy with pharmacological inhibitors (3 methyladenine or bafilomycin) or shRNA knockdown of the autophagic gene Beclin increased tubular cell apoptosis during cisplatin treatment. Our study has found that autophagy occurs in acute kidney injury and this may be an important protective mechanism for cell survival. PMID- 18509316 TI - Angiotensin II activates the Smad pathway during epithelial mesenchymal transdifferentiation. AB - Epithelial to mesenchymal transdifferentiation is a novel mechanism that promotes renal fibrosis and here we investigated whether known causes of renal fibrosis (angiotensin II and transforming growth factor beta, TGFbeta) act through this pathway. We infused angiotensin II into rats for 1 day and found that it activated the Smad pathway which persisted for up to 2 weeks in chronically infused rats. Renal TGF-beta mRNA expression was increased at 3 days and its protein at 2 weeks suggesting Smad pathway activation occurred earlier than TGF beta upregulation. In cultured human tubuloepithelial cells, angiotensin II caused a rapid activation of Smad signaling independent of TGF-beta however, Smad dependent transcription after 1 day was TGF-beta mediated. Two weeks of angiotensin II infusion activated genes associated with epithelial mesenchymal transdifferentiation. Stimulation with angiotensin II for 3 days caused transdifferentiation of the cultured epithelial cells by TGF-beta-mediated processes; however, early changes were independent of endogenous TGF-beta. Smad7 overexpression, which blocks Smad2/3 activation, diminished angiotensin II induced epithelial mesenchymal transdifferentiation. Our results show that angiotensin II activates the Smad signaling system by TGF-beta-independent processes, in vivo and in vitro, causing renal fibrosis. PMID- 18509317 TI - Physician access and early nephrology care in elderly patients with end-stage renal disease. AB - Early nephrology care may improve treatment outcomes of patients with end-stage renal disease. We sought to examine if physician access affects early nephrology care defined as visiting a nephrologist 12 to 4 months before initiating dialysis. The study population consisted of elderly patients starting hemodialysis whose demographic characteristics and initial dialysis therapy were derived from form 2728 files of the Centers for Medicare & Medicaid Services. Early nephrology care, chronic kidney disease and co-morbidities along with access to local non-nephrologist physicians and nephrologists were identified based on Medicare claims and/or United States 2000 Census data. About one-third of elderly patients received early nephrology care prior to initiating dialysis. Patients living in an area with a large number of non-nephrologist physicians or living relatively far away from a nephrologist had a lower likelihood of getting early nephrology care prior to initiating dialysis while those in an area with more practicing nephrologists were more likely to get early nephrology care. The study shows that physician access significantly influences the use of early nephrology care among elderly patients progressing to end-stage renal disease in the United States. PMID- 18509318 TI - Pravastatin improves renal ischemia-reperfusion injury by inhibiting the mevalonate pathway. AB - Statins are known to lessen the severity of renal ischemia-reperfusion injury. The present study was undertaken to define the mechanism of renoprotective actions of statins using a mouse kidney injury model. Treatment of mice with pravastatin, a widely used statin, improved renal function after renal ischemia reperfusion without lowering the plasma cholesterol level. Administration of pravastatin with mevalonate, a product of HMG-CoA reductase, eliminated renal protection suggesting an effect of pravastatin on mevalonate or its metabolism. In hypercholestrolemic apolipoprotein E knockout mice with reduced HMG-CoA reductase activity; the degree of injury was less severe than in control mice, however, there was no protective action of pravastatin on renal injury in the knockout mice. Treatment with a farnesyltransferase inhibitor (L-744832) mimicked pravastatin's protective effect but co-administration with the statin provided no additional protection. Both pravastatin and L-744832 inhibited the injury-induced increase in plasma IL-6 concentration to a similar extent. Our results suggest the protective effect of pravastatin on renal ischemia-reperfusion injury is mediated by inhibition of the mevalonate-isoprenoid pathway independent of its lipid lowering action. PMID- 18509319 TI - Ischemic preconditioning produces systemic protective and adoptively transferable effects. AB - Ischemic preconditioning directly protects organs from subsequent non-specific injuries. To test for systemic protective effects kidneys from F-344 donor rats went through a short warm ischemic time. Both, clamped and contralateral unclamped kidneys were procured after either a short (15 min) or long (24 h) reperfusion period and transplanted into Lewis rats following a prolonged cold ischemia. To test for transferable effects serum from preconditioned rats was infused either into native donors or recipients. Following a short reperfusion interval protective effects were only evident in previously clamped grafts. However, after a long reperfusion interval protective effects were observed in previously clamped and contralateral unclamped kidneys promoting improved survival, structure, function and reduced inflammation. These effects were not related to heme oxygenase-1 induction or neural transmission as heme oxygenase-1 inhibition or denervation prior to preconditioning did not affect organ protection. These results show that renal ischemic preconditioning is associated with time-dependent local and systemically transferable protection. PMID- 18509320 TI - Arteriovenous fistula in the rat tail: a new model of hemodialysis access dysfunction. AB - Problems with vascular access are an important cause of morbidity and mortality in hemodialysis patients. We established a rodent model of arteriovenous fistula by anastomosing the end of a lateral vein to the side of the ventral artery of the rat tail. All operations were technically successful and in all animals the fistula was patent with a dilated fistula vein clearly visible after 28 days. Neointimal hyperplasia was found in 4 out of 5 fistulae with varied pathology from immature to more mature lesions seen both proximal and distal to the anastomosis. There was no particular pattern to the presence of or type of lesion found at any particular site of the fistulae. This fistula promises to be useful in analyzing pathologic processes that occur in native arteriovenous fistulae since the vein is accessible to functional studies and to test new subcutaneous or intravascular treatments. PMID- 18509321 TI - Urinary exosomal transcription factors, a new class of biomarkers for renal disease. AB - Urinary exosomes are excreted from all nephron segments and constitute a rich source of intracellular kidney injury biomarkers. To study whether they contain transcription factors, we collected urine from two acute kidney injury models (cisplatin or ischemia-reperfusion), two podocyte injury models (puromycin treated rats or podocin-Vpr transgenic mice) and from patients with focal segmental glomerulosclerosis, acute kidney injury and matched controls. Exosomes were isolated by differential centrifugation and found to contain activating transcription factor 3 (ATF3) and Wilms Tumor 1 (WT-1) proteins detected by Western blot. These factors were found in the concentrated exosomal fraction, but not in whole urine. ATF3 was continuously present in urine exosomes of the rat models following acute injury at times earlier than the increase in serum creatinine. ATF3 was found in exosomes isolated from patients with acute kidney injury but not from patients with chronic kidney disease or controls. Urinary WT 1 was present in animal models before significant glomerular sclerosis and in 9/10 patients with focal segmental glomerulosclerosis but not in 8 controls. Our findings suggest that transcription factor ATF3 may provide a novel renal tubular cell biomarker for acute kidney injury while WT-1 may detect early podocyte injury. Measurement of urinary exosomal transcription factors may offer insight into cellular regulatory pathways. PMID- 18509322 TI - Triptolide protects podocytes from puromycin aminonucleoside induced injury in vivo and in vitro. AB - Extracts of Tripterygium wilfordii Hook F have been used to treat glomerulonephritis for more than 30 years in China with dramatic antiproteinuric effects. Triptolide, a diterpene triepoxide, is one of the major active components of these extracts. To clarify its antiproteinuric effects we induced podocyte injury by puromycin aminonucleoside. Triptolide effectively reduced the proteinuria induced by puromycin in nephrotic rats without reducing the glomerular filtration rate. The antiproteinuric effect was associated with improvement in the foot process effacement, a decrease in the podocyte injury marker desmin as well as the restoration of nephrin and podocin expression and distribution. In cultured mouse podocytes triptolide pretreatment prevented the puromycin-induced disruption of the actin cytoskeleton and microfilament associated synaptopodin while protecting nephrin and podocin expression. Triptolide suppressed reactive oxygen species generation and p38 mitogen activated protein kinase activation while restoring RhoA signaling activity. These results show that triptolide ameliorates puromycin aminonucleoside-mediated podocyte injury in vivo and in vitro. PMID- 18509323 TI - Effect of apelin on glomerular hemodynamic function in the rat kidney. AB - Apelin is a vasoactive peptide identified as the endogenous ligand of an orphan G protein-coupled receptor called APJ. Apelin and its receptor have been found in the brain and the cardiovascular system. Here we show that the apelin receptor mRNA is highly expressed in the glomeruli while its level of expression is lower in all nephron segments including collecting ducts that express vasopressin V2 receptors. Intravenous injection of apelin 17 into lactating rats induced a significant diuresis. Apelin receptor mRNA was also found in endothelial and vascular smooth muscle cells of glomerular arterioles. Apelin administration caused vasorelaxation of angiotensin II-preconstricted efferent and afferent arterioles as shown by an increase in their diameter. Activation of endothelial apelin receptors caused release of nitric oxide which inhibited angiotensin II induced rise in intracellular calcium. In addition, it appears that apelin had a direct receptor-mediated vasoconstrictive effect on vascular smooth muscle. These results show that apelin has complex effects on the pre- and post glomerular microvasculature regulating renal hemodynamics. Its role on tubular function (if any) remains to be determined. PMID- 18509324 TI - Renal safety of annual zoledronic acid infusions in osteoporotic postmenopausal women. AB - Intravenous bisphosphonates reduce fracture risk but have been associated in rare cases with deteriorating renal-function in cancer patients. The renal effects of zoledronic acid were assessed in osteoporotic postmenopausal women from 27 countries who received three annual infusions of zoledronic acid or a placebo in a randomized, double-blind trial. Serum creatinine, estimated creatinine clearance and urinary protein were measured before and after at least one infusion in a predefined renal safety cohort of 5035 equally divided patients. This group was compared to 7714 patients whose parameters were measured annually. Significantly more transient pre- to post-infusion increases in serum creatinine occurred in zoledronic acid than placebo-treated patients with significant elevations, relative to pre-infusion, only in the second year. All 31 zoledronic acid and 8 of 10 patients on placebo recovered their pre-infusion serum creatinine value within 12 months. No differences in mean changes in serum creatinine, estimated creatinine clearance or adverse renal events were found. We found that transient changes in renal function can occur following an annual zoledronic acid infusion but, in the long term, renal function was not different from control patients. PMID- 18509325 TI - Kt/V underestimates the hemodialysis dose in women and small men. AB - Current guidelines suggest a minimum Kt/V of 1.2 for three weekly hemodialysis sessions; however, using V as a normalizing factor has been questioned. Parameters such as weight(0.67) (W(0.67)) and body surface area (BSA) that reflect the metabolic rate may be preferable. To determine this, we studied 328 hemodialysis patients (221 male) with a target Kt/V of 1.2. Using this relationship and the individual's Watson Volume, we calculated the Kt, Kt/BSA, and Kt/W(0.67) equivalent to the target and measured the effects of body size and gender on these parameters for each patient. The target corresponded to a range of equivalent Kt/BSA and Kt/W(0.67) each significantly higher in males than females and in larger than smaller males. V/BSA and V/W(0.67), the conversion factors of Kt/V to Kt/BSA and Kt/W(0.67) respectively, were significantly greater in males than females and heavier than lighter men. Our study shows that if Kt/BSA and Kt/W(0.67) reflect the true required dose, prescribing a target Kt/V of 1.2 would underestimate this in females and in small males. Further work is required to develop clinical outcome-based adequacy targets. PMID- 18509326 TI - Adverse drug reactions with halogenated anesthetics. AB - Problems with organ toxicity from the halogenated volatile anesthetics have influenced clinical practice, anesthetic selection, and drug development for half a century.(1,2) In turn, investigations that elucidated the mechanisms of volatile anesthetics adverse effects identified new mechanisms, provided remarkable insights into human toxicology, and represented seminal contributions to clinical pharmacology. This review focuses on adverse organ effects (hepatic, renal, and others) that are attributable to anesthetic metabolism and/or degradation. Routine reversible effects on pulmonary, cardiac, and other organ functions are not addressed. PMID- 18509327 TI - Pharmacogenetic pathway analysis of docetaxel elimination. AB - The purpose of this study was to evaluate the affinity of docetaxel for 14 transporter proteins and assess the functional significance of 17 variants in five genes involved in drug elimination. Among the transfected models investigated, OATP1B3 (SLCO1B3) was identified as the most efficient influx transporter for docetaxel. None of the observed genotypes (SLCO1B3, ABCB1, and ABCC2) was related with docetaxel clearance in 92 white patients (P > 0.17). However, the simultaneous presence of the CYP3A4*1B and CYP3A5*1A alleles was associated with a 64% increase in docetaxel clearance (P = 0.0015), independent of both sex and CYP3A activity (as determined using the erythromycin breath test). This haplotype was also associated with increased midazolam clearance in another population (P = 0.0198). An analysis of the CYP3A locus among CEPH-HapMap samples revealed that CYP3A4*1B is present exclusively among a subset of CYP3A5 expressors. Therefore, future studies should first stratify the population on the basis of CYP3A5 genotype and then compare CYP3A activity between individuals with and without the CYP3A4*1B allele. PMID- 18509328 TI - Influence of solute carriers on the pharmacokinetics of CYP3A4 probes. AB - We hypothesized that the assessment of baseline CYP3A4 activity is influenced by probe-specific differences in hepatocellular uptake mechanisms. There was no significant correlation between the erythromycin breath test (ERMBT) parameters and midazolam clearance in 30 cancer patients (R(2) < 0.01), regardless of their CYP3A5 genotype status. In cellular models overexpressing 10 different solute carriers, erythromycin uptake was significantly increased by OATP1A2 (P < 0.005) and OATP1B3 (P < 0.01). Midazolam was not a substrate for any of the tested transporters. In a separate cohort of 119 patients, 6 nonsynonymous variants in the OATP1B3 gene SLCO1B3 were identified. Individuals carrying two copies of the T allele at the 334 locus had a 2.4-fold lower value for ERMBT 1/T(max) (P = 0.001), a measure reflecting more rapid hepatic uptake. These findings suggest that differential affinities for solute carriers should be considered when selecting an appropriate phenotypic probe to allow tailored dosing of pharmaceuticals that are CYP3A4 substrates. PMID- 18509329 TI - Etoposide sensitivity does not predict MLL rearrangements or risk of therapy related acute myeloid leukemia. AB - Therapy-related acute myeloid leukemia (t-AML) caused by MLL rearrangements (rMLL) can arise from topoisomerase II agents. However, whether rMLL-related leukemogenesis is inextricably linked to drug cytotoxicity remains controversial. We therefore compared (i) rMLL in children with acute lymphoblastic leukemia (ALL) who developed t-AML and those who did not, (ii) epipodophyllotoxin toxicity in patients with t-AML and in controls, and (iii) rMLL in cells sensitive to etoposide and in those resistant to etoposide. In children with ALL, rMLL appeared to be more frequent in children who developed t-AML than in those who did not (seven pairs, P = 0.04), although independent of the cumulative etoposide dose (P = 0.5). Similarly, the frequency of epipodophyllotoxin-related toxicities did not differ between patients with t-AML and controls (26 pairs, P > 0.17). Moreover, in 25 cell lines, etoposide-induced MLL fusions did not differ in sensitive vs. resistant lines at equitoxic concentrations (P = 0.65). Together, these results indicate that epipodophyllotoxin-mediated leukemogenesis is not directly linked to drug cytotoxicity. PMID- 18509330 TI - Disruptive effect of midazolam on fear memory reconsolidation: decisive influence of reactivation time span and memory age. AB - Benzodiazepine (BDZ) administered shortly after retrieval disrupts the reconsolidation of fear memory. In this research, we explored the way in which different factors that limit the emergence of such process may affect BDZ's disruptive effect on fear memory reconsolidation. Animals were conditioned in a contextual fear paradigm; the consolidated memory was reactivated by exposure to the associated context for different periods of time that were followed by midazolam (MDZ) administration. We also studied MDZ amnesic effect after reactivating fear memories of several ages. We finally analyzed the effectiveness of different MDZ doses in preventing the reconsolidation of different age fear memories. The memory trace was disrupted following MDZ when the reactivation session lasted 3-5 min but it was not after a briefer 1-min reactivation period. Over a 10-min reactivation session, all animals gradually reduced their fear response, which indicates the emergence of the extinction process. When tested, MDZ rats exhibited a robust fear, suggesting that MDZ impaired the consolidation of extinction. In a 3-min reactivation session, MDZ (1-1.5 mg/kg) prevented the reconsolidation of recently acquired memories. A 21-day-old fear memory was only vulnerable to MDZ at a 1.5 mg/kg dose with a reactivation session of 5 and not 3 min, whereas a 36-day-old memory was only disrupted with a higher MDZ dose (3 mg/kg) regardless of the reactivation trial's duration. This study demonstrated MDZ's interference on fear-memory reconsolidation within a relatively short reactivation period in recently acquired memories. Over longer reexposure, MDZ disrupts the consolidation of extinction. A longer duration of the reexposure session, as well as higher MDZ doses, is required to prevent the reconsolidation process of remote fear memories. PMID- 18509331 TI - Hydrogen sulphide release to surface waters at the Precambrian/Cambrian boundary. AB - Animal-like multicellular fossils appeared towards the end of the Precambrian, followed by a rapid increase in the abundance and diversity of fossils during the Early Cambrian period, an event also known as the 'Cambrian explosion'. Changes in the environmental conditions at the Precambrian/Cambrian transition (about 542 Myr ago) have been suggested as a possible explanation for this event, but are still a matter of debate. Here we report molybdenum isotope signatures of black shales from two stratigraphically correlated sample sets with a depositional age of around 542 Myr. We find a transient molybdenum isotope signal immediately after the Precambrian/Cambrian transition. Using a box model of the oceanic molybdenum cycle, we find that intense upwelling of hydrogen sulphide-rich deep ocean water best explains the observed Early Cambrian molybdenum isotope signal. Our findings suggest that the Early Cambrian animal radiation may have been triggered by a major change in ocean circulation, terminating a long period during which the Proterozoic ocean was stratified, with sulphidic deep water. PMID- 18509332 TI - Two-band superconductivity in LaFeAsO0.89F0.11 at very high magnetic fields. AB - The recent synthesis of the superconductor LaFeAsO(0.89)F(0.11) with transition temperature T(c) approximately 26 K (refs 1-4) has been quickly followed by reports of even higher transition temperatures in related compounds: 41 K in CeFeAsO(0.84)F(0.16) (ref. 5), 43 K in SmFeAsO(0.9)F(0.1) (ref. 6), and 52 K in NdFeAsO(0.89)F(0.11) and PrFeAsO(0.89)F(0.11) (refs 7, 8). These discoveries have generated much interest in the mechanisms and manifestations of unconventional superconductivity in the family of doped quaternary layered oxypnictides LnOTMPn (Ln: La, Pr, Ce, Sm; TM: Mn, Fe, Co, Ni; Pn: P, As), because many features of these materials set them apart from other known superconductors. Here we report resistance measurements of LaFeAsO(0.89)F(0.11) at high magnetic fields, up to 45 T, that show a remarkable enhancement of the upper critical field B(c2) compared to values expected from the slopes dB(c2)/dT approximately 2 T K(-1) near T(c), particularly at low temperatures where the deduced B(c2)(0) approximately 63-65 T exceeds the paramagnetic limit. We argue that oxypnictides represent a new class of high-field superconductors with B(c2) values surpassing those of Nb(3)Sn, MgB(2) and the Chevrel phases, and perhaps exceeding the 100 T magnetic field benchmark of the high-T(c) copper oxides. PMID- 18509333 TI - Magnetic order close to superconductivity in the iron-based layered LaO1-xFxFeAs systems. AB - Following the discovery of long-range antiferromagnetic order in the parent compounds of high-transition-temperature (high-T(c)) copper oxides, there have been efforts to understand the role of magnetism in the superconductivity that occurs when mobile 'electrons' or 'holes' are doped into the antiferromagnetic parent compounds. Superconductivity in the newly discovered rare-earth iron-based oxide systems ROFeAs (R, rare-earth metal) also arises from either electron or hole doping of their non-superconducting parent compounds. The parent material LaOFeAs is metallic but shows anomalies near 150 K in both resistivity and d.c. magnetic susceptibility. Although optical conductivity and theoretical calculations suggest that LaOFeAs exhibits a spin-density-wave (SDW) instability that is suppressed by doping with electrons to induce superconductivity, there has been no direct evidence of SDW order. Here we report neutron-scattering experiments that demonstrate that LaOFeAs undergoes an abrupt structural distortion below 155 K, changing the symmetry from tetragonal (space group P4/nmm) to monoclinic (space group P112/n) at low temperatures, and then, at approximately 137 K, develops long-range SDW-type antiferromagnetic order with a small moment but simple magnetic structure. Doping the system with fluorine suppresses both the magnetic order and the structural distortion in favour of superconductivity. Therefore, like high-T(c) copper oxides, the superconducting regime in these iron-based materials occurs in close proximity to a long-range ordered antiferromagnetic ground state. PMID- 18509334 TI - Dissecting direct reprogramming through integrative genomic analysis. AB - Somatic cells can be reprogrammed to a pluripotent state through the ectopic expression of defined transcription factors. Understanding the mechanism and kinetics of this transformation may shed light on the nature of developmental potency and suggest strategies with improved efficiency or safety. Here we report an integrative genomic analysis of reprogramming of mouse fibroblasts and B lymphocytes. Lineage-committed cells show a complex response to the ectopic expression involving induction of genes downstream of individual reprogramming factors. Fully reprogrammed cells show gene expression and epigenetic states that are highly similar to embryonic stem cells. In contrast, stable partially reprogrammed cell lines show reactivation of a distinctive subset of stem-cell related genes, incomplete repression of lineage-specifying transcription factors, and DNA hypermethylation at pluripotency-related loci. These observations suggest that some cells may become trapped in partially reprogrammed states owing to incomplete repression of transcription factors, and that DNA de-methylation is an inefficient step in the transition to pluripotency. We demonstrate that RNA inhibition of transcription factors can facilitate reprogramming, and that treatment with DNA methyltransferase inhibitors can improve the overall efficiency of the reprogramming process. PMID- 18509335 TI - Magnetic resonance imaging of pH in vivo using hyperpolarized 13C-labelled bicarbonate. AB - As alterations in tissue pH underlie many pathological processes, the capability to image tissue pH in the clinic could offer new ways of detecting disease and response to treatment. Dynamic nuclear polarization is an emerging technique for substantially increasing the sensitivity of magnetic resonance imaging experiments. Here we show that tissue pH can be imaged in vivo from the ratio of the signal intensities of hyperpolarized bicarbonate (H(13)CO(3)(-)) and (13)CO(2) following intravenous injection of hyperpolarized H(13)CO(3)(-). The technique was demonstrated in a mouse tumour model, which showed that the average tumour interstitial pH was significantly lower than the surrounding tissue. Given that bicarbonate is an endogenous molecule that can be infused in relatively high concentrations into patients, we propose that this technique could be used clinically to image pathological processes that are associated with alterations in tissue pH, such as cancer, ischaemia and inflammation. PMID- 18509336 TI - Analysis of a spatial orientation memory in Drosophila. AB - Flexible goal-driven orientation requires that the position of a target be stored, especially in case the target moves out of sight. The capability to retain, recall and integrate such positional information into guiding behaviour has been summarized under the term spatial working memory. This kind of memory contains specific details of the presence that are not necessarily part of a long term memory. Neurophysiological studies in primates indicate that sustained activity of neurons encodes the sensory information even though the object is no longer present. Furthermore they suggest that dopamine transmits the respective input to the prefrontal cortex, and simultaneous suppression by GABA spatially restricts this neuronal activity. Here we show that Drosophila melanogaster possesses a similar spatial memory during locomotion. Using a new detour setup, we show that flies can remember the position of an object for several seconds after it has been removed from their environment. In this setup, flies are temporarily lured away from the direction towards their hidden target, yet they are thereafter able to aim for their former target. Furthermore, we find that the GABAergic (stainable with antibodies against GABA) ring neurons of the ellipsoid body in the central brain are necessary and their plasticity is sufficient for a functional spatial orientation memory in flies. We also find that the protein kinase S6KII (ignorant) is required in a distinct subset of ring neurons to display this memory. Conditional expression of S6KII in these neurons only in adults can restore the loss of the orientation memory of the ignorant mutant. The S6KII signalling pathway therefore seems to be acutely required in the ring neurons for spatial orientation memory in flies. PMID- 18509337 TI - Cortical control of a prosthetic arm for self-feeding. AB - Arm movement is well represented in populations of neurons recorded from the motor cortex. Cortical activity patterns have been used in the new field of brain machine interfaces to show how cursors on computer displays can be moved in two- and three-dimensional space. Although the ability to move a cursor can be useful in its own right, this technology could be applied to restore arm and hand function for amputees and paralysed persons. However, the use of cortical signals to control a multi-jointed prosthetic device for direct real-time interaction with the physical environment ('embodiment') has not been demonstrated. Here we describe a system that permits embodied prosthetic control; we show how monkeys (Macaca mulatta) use their motor cortical activity to control a mechanized arm replica in a self-feeding task. In addition to the three dimensions of movement, the subjects' cortical signals also proportionally controlled a gripper on the end of the arm. Owing to the physical interaction between the monkey, the robotic arm and objects in the workspace, this new task presented a higher level of difficulty than previous virtual (cursor-control) experiments. Apart from an example of simple one-dimensional control, previous experiments have lacked physical interaction even in cases where a robotic arm or hand was included in the control loop, because the subjects did not use it to interact with physical objects-an interaction that cannot be fully simulated. This demonstration of multi-degree-of-freedom embodied prosthetic control paves the way towards the development of dexterous prosthetic devices that could ultimately achieve arm and hand function at a near-natural level. PMID- 18509338 TI - Induced ncRNAs allosterically modify RNA-binding proteins in cis to inhibit transcription. AB - With the recent recognition of non-coding RNAs (ncRNAs) flanking many genes, a central issue is to obtain a full understanding of their potential roles in regulated gene transcription programmes, possibly through different mechanisms. Here we show that an RNA-binding protein, TLS (for translocated in liposarcoma), serves as a key transcriptional regulatory sensor of DNA damage signals that, on the basis of its allosteric modulation by RNA, specifically binds to and inhibits CREB-binding protein (CBP) and p300 histone acetyltransferase activities on a repressed gene target, cyclin D1 (CCND1) in human cell lines. Recruitment of TLS to the CCND1 promoter to cause gene-specific repression is directed by single stranded, low-copy-number ncRNA transcripts tethered to the 5' regulatory regions of CCND1 that are induced in response to DNA damage signals. Our data suggest that signal-induced ncRNAs localized to regulatory regions of transcription units can act cooperatively as selective ligands, recruiting and modulating the activities of distinct classes of RNA-binding co-regulators in response to specific signals, providing an unexpected ncRNA/RNA-binding protein-based strategy to integrate transcriptional programmes. PMID- 18509339 TI - Neuroscience: Brain control of a helping hand. PMID- 18509340 TI - Central mechanisms of osmosensation and systemic osmoregulation. AB - Systemic osmoregulation is a vital process whereby changes in plasma osmolality, detected by osmoreceptors, modulate ingestive behaviour, sympathetic outflow and renal function to stabilize the tonicity and volume of the extracellular fluid. Furthermore, changes in the central processing of osmosensory signals are likely to affect the hydro-mineral balance and other related aspects of homeostasis, including thermoregulation and cardiovascular balance. Surprisingly little is known about how the brain orchestrates these responses. Here, recent advances in our understanding of the molecular, cellular and network mechanisms that mediate the central control of osmotic homeostasis in mammals are reviewed. PMID- 18509341 TI - KIR2DL5 alleles mark certain combination of activating KIR genes. AB - Killer cell Ig-like receptors (KIR) control the immune response of NK cells and some T cells to infections and tumors. KIR genes evolve rapidly and are variable between individuals in their number, type and sequence. Here, we determined the nature of KIR2DL5 gene polymorphism in four ethnic groups using direct DNA sequencing method. Nine new sequences were discovered. Within the panel of 248 KIR2DL5-positive individuals, 14 KIR2DL5-sequences differing in coding regions were observed. They differed at only seven amino acid positions, and such limited polymorphism is consistent with its conserved nature throughout the hominoid lineage. Ethnic deviation was seen in the distribution of KIR2DL5A, KIR2DL5B and their alleles. African Americans had more KIR2DL5 alleles than other populations indicating that more polymorphisms are yet to be discovered in Africans. Linkage between KIR2DL5-alleles and certain activating-KIR genes were observed, but frequency of these linked clusters differed substantially between populations. Consequently, KIR2DL5 alleles can be used as markers to predict the activating KIR gene content. Typing system distinguishing A*001 and B*002 alleles can serve as a powerful screening test to assess the content of most variable activating KIR genes that have been implicated in human disease and in the outcome of hematopoietic stem cell transplantation. PMID- 18509342 TI - Targeting the arterial wall, but what is the target? PMID- 18509343 TI - A modified ambulatory arterial stiffness index is independently associated with all-cause mortality. AB - Dependence of the ambulatory arterial stiffness index (AASI) on data scattering interferes with its potential clinical relevance. We assessed the correlates and all-cause mortality associations of a modified AASI (s-AASI). AASI was derived from the 24-h diastolic vs. systolic blood pressure linear regression line, whereas s-AASI was derived by symmetric regression (bisecting the line of diastolic vs systolic and systolic vs. diastolic). Of 2918 patients 55% were women; age was 56 +/- 16 years and body mass index was 27.3 +/- 4.5 kg/m(2). Average 24-h ambulatory blood pressure was 138 +/- 16/78 +/- 10 mm Hg. Applying the modified method for calculating AASI yielded a different measure: the negative correlation between AASI and blood pressure dipping (r = -0.304, P < 0.0001) was abolished (r = +0.223, P < 0.0001), s-AASI was more dependent on age (r = 0.266 vs. r = 0.089 for AASI), and prediction of all-cause mortality was enhanced; hazard ratio (95% confidence intervals) 1.17 (1.00-1.36) per 1 s.d. increase in s-AASI in the fully adjusted model as compared with 1.15 (0.97-1.36) for AASI. PMID- 18509344 TI - Arterial stiffness: beyond pulse wave velocity and its measurement. PMID- 18509345 TI - Antihypertensive drug class and dyslipidemia: risk association among Chinese patients with uncomplicated hypertension. PMID- 18509346 TI - Macro and microvasculature in hypertension: therapeutic aspects. AB - Macrovasculature, microvasculature and the heart determine the structure and function of the circulatory system. Due to the viscoelastic properties of large arteries, the pulsatile pressure and flow that result from intermittent ventricular ejection is smoothed out, so that microvasculature mediates the delivery of nutrients and oxygen to tissues steadily. The disruption of this function, which occurs when microvascular structure develops in response to hypertension, leads to end-organ damage. Microvascular structure is not only the site of vascular resistance, but also the origin of most of the wave reflections generating increased central systolic blood pressure (SBP) in the elderly. Nowadays many data of the literature suggest that hypertension-related damage to the micro and macrovascular system may be manageable through pharmacological agents. Among them, beta-blocking agents and diuretics poorly modify microvascular structure, whereas angiotensin and calcium entry blockade has an opposite effect, thereby reducing central wave reflections and, finally, causing a selective SBP reduction. PMID- 18509347 TI - Predictors of new-onset diabetes mellitus in hypertensive patients: the VALUE trial. AB - Diabetes mellitus often develops in patients with hypertension. We investigated predictors of diabetes mellitus development in hypertensives at risk of developing the disease in the VALUE trial population. Among the 9995 non-diabetic hypertensive patients at baseline, 1298 patients developed diabetes mellitus during the average follow-up of 4.2 years. New-onset diabetes mellitus was defined from adverse event reports, information about new antidiabetic medication and/or a fasting glucose >or=7.0 mmol l(-1) at the end of trial. Twenty-five potential baseline predictors of new-onset diabetes mellitus were analysed by univariate logistic regression and 14 of 25 predictors were found to be statistically significant with a P-value <0.05. The predictors were in order of decreasing significance; glucose, body mass index (BMI), age, uric acid, non Caucasian race, haemoglobin, heart rate, randomized study treatment, history of coronary heart disease (CHD), gender, total cholesterol, proteinuria, potassium and creatinine. Multivariate stepwise logistic regression analyses were used and potential baseline predictors of new-onset diabetes mellitus were considered significant by four different models (P-value <0.001). The final multivariate model selected included all patients, but not treatment group as a potential predictor, and the six significant predictors identified from this model were glucose, BMI, non-Caucasian race, age, heart rate and history of CHD. In conclusion, glucose and BMI were the most important predictors of new-onset diabetes mellitus in hypertensive patients at high cardiovascular risk, and easily accessible clinical characteristics strongly predict patients at risk of developing diabetes mellitus. PMID- 18509348 TI - The prognostic value of haemodynamic parameters in the recovery phase of an exercise test. The Finnish Cardiovascular Study. AB - We tested the hypothesis that the change from the peak to recovery values of systolic arterial pressure (SAP recovery) and rate-pressure product (RPP recovery) can be used to predict all-cause and cardiovascular mortality, as well as sudden cardiac death (SCD) in patients referred to a clinical exercise stress test. As a part of the Finnish Cardiovascular Study (FINCAVAS), consecutive patients (n=2029; mean age+/-SD=57+/-13 years; 1290 men and 739 women) with a clinically indicated exercise test using a bicycle ergometer were included in the present study. Capacities of attenuated SAP recovery, RPP recovery and heart rate recovery (HRR) to stratify the risk of death were estimated. During a follow-up (mean+/-s.d.) of 47+/-13 months, 122 patients died; 58 of the deaths were cardiovascular and 33 were SCD. In Cox regression analysis after adjustment for the peak level of the variable under assessment, age, sex, use of beta-blockers, previous myocardial infarction and other common coronary risk factors, the hazard ratio of the continuous variable RPP recovery (in units 1000 mm Hg x b.p.m.) was 0.85 (95% CI: 0.73-0.98) for SCD, 0.87 (0.78-0.97) for cardiovascular mortality, and 0.87 (0.81 to 0.94) for all-cause mortality. SAP recovery was not a predictor of mortality. The relative risks of having HRR below 18 b.p.m., a widely used cutoff point, were as follows: for SCD 1.28 (0.59-2.81, ns), for cardiovascular mortality 2.39 (1.34-4.26) and for all-cause mortality 2.40 (1.61-3.58). In conclusion, as a readily available parameter, RPP recovery is a promising candidate for a prognostic marker. PMID- 18509349 TI - Noise pollution in hemodialysis centers. PMID- 18509350 TI - 5-Azacitidine has limited therapeutic activity in myelofibrosis. PMID- 18509351 TI - Aberrant expression of IL-22 receptor 1 and autocrine IL-22 stimulation contribute to tumorigenicity in ALK+ anaplastic large cell lymphoma. AB - One of the characteristic features of anaplastic lymphoma kinase (ALK)(+), anaplastic large cell lymphoma (ALK(+)ALCL) is the constitutive activation of signal transducers and activators of transcription-3 (STAT3), a defect believed to be important for the pathogenesis of these tumors. In this report, we describe the existence of an autocrine stimulatory loop involving interleukin-22 (IL-22) that contributes to STAT3 activation and tumorigenicity of ALK(+)ALCL. The IL-22 receptor, a heterodimer composed of IL-22R1 and IL-10R2, was expressed in all ALK(+)ALCL cell lines and tumors examined. The expression of IL-22R1 in ALK(+)ALCL is aberrant, as this protein is absent in benign lymphocytes. Although ALK(+)ALCL cells produce endogenous IL-22, addition of recombinant IL-22 to ALK(+)ALCL cell lines significantly increased STAT3 activation, cell proliferation and colony formation in soft agar. Opposite biological effects were observed in cells treated with recombinant IL-22 binding protein (a naturally occurring IL-22 decoy) or IL-22-neutralizing antibody. Nucleophosmin (NPM)-ALK, the characteristic fusion gene oncoprotein expressed in ALK(+)ALCL, directly contributes to the aberrant expression of IL-22R1, as transfection of NPM-ALK in Jurkat cells-induced IL-22R1 expression and IL-22-mediated STAT3 activation. To conclude, for the first time, we demonstrate the importance of the IL-22 autocrine pathway in a lymphoid malignancy, and reveal yet another novel function of NPM-ALK. PMID- 18509353 TI - Resistance to FLT3 inhibition in an in vitro model of primary AML cells with a stem cell phenotype in a defined microenvironment. AB - Relapse in acute myeloid leukaemia (AML) is mediated by survival of leukaemic stem cells following remission-induction chemotherapy. It would therefore be useful to identify therapeutic agents that target leukaemic stem cells. We devised a flow cytometric chemosensitivity assay allowing 48 h culture of leukaemic blasts in a defined microenvironment followed by enumeration of viable CD34+CD38-CD123+ leukaemic stem and progenitor cells (LSPC). The assay was used to investigate the LSPC response to cytosine arabinoside (Ara-C) and to the FLT3 inhibitor AG1296. There was a 3.6-fold increase in Ara-C-treated LSPC survival under defined 'niche-like' conditions compared to culture without microenvironmental support. Nine AML samples with internal tandem duplications of FLT3 (FLT3/ITDs) were treated with AG1296. Three samples were very sensitive (>50% kill) and 4 were moderately sensitive (10-50% kill) in bulk suspension culture without microenvironmental support. However, under defined 'niche-like' conditions, the survival of LSPC was enhanced rather than inhibited by AG1296 treatment. We conclude that an interaction between LSPC and a defined in vitro microenvironment models a chemoresistant niche. Our data point to a need to investigate more novel chemotherapeutic agents under these stringent conditions to identify agents that may be suitable to target minimal residual disease in AML. PMID- 18509352 TI - The role of vertebral augmentation in multiple myeloma: International Myeloma Working Group Consensus Statement. PMID- 18509354 TI - An activating KRAS mutation in imatinib-resistant chronic myeloid leukemia. PMID- 18509355 TI - Treatment of plasma cell dyscrasias with lenalidomide. AB - Lenalidomide is an immunomodulatory drug, structurally related to thalidomide, with pleiotropic activity including antiangiogenic and antineoplastic properties. It is the product of advances in our understanding of the biology of myeloma cells, their interactions with the microenvironment and of the underlying molecular pathways. In preclinical and clinical studies, lenalidomide was more potent and less toxic than thalidomide. Subsequent phase II and III studies confirmed the activity of lenalidomide either as a single agent or in combination with dexamethasone in relapsed or refractory myeloma patients, whereas combinations with chemotherapy induce high response rates and durable remissions. Lenalidomide has been used successfully as an upfront treatment either with high or low dose dexamethasone or with melphalan and prednisone, resulting in high overall response and complete response rates and excellent 1-year survival. Lenalidomide causes less neuropathy than thalidomide; however, the risk of thromboembolism is high, especially in patients treated with lenalidomide and steroids. In this review, we summarize the mechanisms of action, toxicity and clinical activity, and the current role of lenalidomide in patients with multiple myeloma or other related plasma cell disorders. PMID- 18509356 TI - Oncolytic virotherapy for multiple myeloma using a tumour-specific double-deleted vaccinia virus. PMID- 18509357 TI - Inactive and highly active, proteolytically processed transglutaminase-5 in epithelial cells. AB - Transglutaminases (TGs) are Ca(2+)-dependent enzymes capable of catalyzing transamidation of glutamine residues to form intermolecular isopeptide bonds. These enzymes are involved in various biological phenomena, including blood coagulation, wound healing, cell death, tissue repair, and terminal differentiation of keratinocytes. Among the TG-family members, TG5 is one of the latest identified enzymes and therefore the less characterized at the functional level. In this work, we reported that TG5 is proteolytically processed in the baculovirus expression system and in mammal epithelial cells. Similar to other members of the TG family-TG1, TG3, and factor XIIIa -, TG5 full-length enzyme has very low enzymatic activity, while the 53-kDa proteolytically processed form is highly active. PMID- 18509358 TI - X-chromosome inactivation and skin disease. AB - X-chromosome inactivation (XCI) is the process in which females transcriptionally silence one of their two X chromosomes in early embryonic development, equalizing X chromosome gene expression between males and females. XCI depends on a gene called XIST, a functional RNA molecule that does not code for a protein. Recent studies indicate abundant intergenic transcription and nonprotein coding RNAs in the human genome, which are suspected to function in modulating gene expression. XCI may therefore serve as a useful model to learn and understand the potential function of these elements, as well as their effects on human disease. Here, we review the genetic and molecular basis of XCI and describe how the mechanistics of this process lead to the phenotypes of X-linked skin diseases, most notably in the pattern of lines, swirls, and whorls first noted by the dermatologist Alfred Blaschko. We suggest that XCI, and other epigenetic phenomena, will continue to impact our understanding of the genetic mechanisms of disease.Journal of Investigative Dermatology (2008) 128, 2753-2759; doi:10.1038/jid.2008.145; published online 29 May 2008. PMID- 18509359 TI - IL-12 deficiency exacerbates inflammatory responses in UV-irradiated skin and skin tumors. AB - IL-12 deficiency has been shown to promote photocarcinogenesis in mice. As UVB induced inflammation is an important tumor-promoting event in the development of skin tumors, we determined the effects of IL-12-deficiency on UVB-induced inflammatory responses in mice. For this purpose, IL-12-knockout (IL-12 KO) and their wild-type counterparts were subjected to a photocarcinogenesis protocol; skin and tumor samples were collected at the termination of the experiment, and analyzed for biomarkers of inflammation and their mediators. We found that the levels of infiltrating leukocytes, myeloperoxidase, proliferating cell-nuclear antigen (PCNA), COX-2, PGE2, and the proinflammatory cytokines IL-1beta, TNF alpha, and IL-6 were higher in the UVB-exposed skin of IL-12 KO than in that of wild-type mice. In a short-term experiment, pretreatment of IL-12 KO mice with rIL-12 (50 ng per mouse) before each exposure to UVB increased the repair rate of UVB-induced cyclobutane pyrimidine dimers, while inhibiting UVB-induced increases in myeloperoxidase, COX-2, PGE2, PCNA, TNF-alpha, and IL-1beta in the skin as compared with non-rIL-12-treated IL-12 KO mice. Similarly, tumors of IL-12 KO mice expressed higher levels of inflammatory responses than those of wild-type mice. Together, our data suggest that IL-12 KO mice are more susceptible to both UVB-induced inflammation and photocarcinogenesis because of the deficiency in the repair of UVB-induced DNA damage. PMID- 18509360 TI - Skin-infiltrating monocytes/macrophages migrate to draining lymph nodes and produce IL-10 after contact sensitizer exposure to UV-irradiated skin. AB - Low-dose UVB exposure induces antigen-specific unresponsiveness to antigen(s) introduced through UV-irradiated skin (tolerance). Analysis of cytokine expression in murine draining lymph nodes (DLNs) revealed that IL-12p40 mRNA and protein expression as well as IL-12p70 protein were upregulated after application of the contact sensitizer 2,4 dinitro-1-fluorobenzene (DNFB) to normal skin. The cellular source of IL-12p40 mRNA was CD11c+ cells. By contrast, following DNFB application to UV-irradiated skin (UV+DNFB), IL-12p40 mRNA was not upregulated, and DLN IL-12p40 and p70 proteins were reduced. UVB irradiation alone did not upregulate IL-10 mRNA, but UV+DNFB upregulated IL-10 mRNA as early as 3-6 hours after DNFB application, immediately preceding a decrease of IL-12p40 mRNA from the level induced by UVB. The infiltration of F4/80+ cells into UV-irradiated skin was followed by a rapid and remarkable increase of F4/80+CD11c(-) cells in DLN 3 hours following DNFB application. FITC/DNFB skin painting and subsequent enzyme-linked immunospot assay demonstrated that flow-sorted FITC+F4/80+CD11c(-) cells from the DLN produce IL-10. Thus, monocytes/macrophages that infiltrated into the skin following UVB exposure migrate to the DLN triggered by contact sensitizers. Production of IL-10 by migrating macrophages, in conjunction with IL 12 inhibition in the DLN, likely reflects a role as mobile suppressive mediators for locally induced UV tolerance. PMID- 18509361 TI - Lack of cytoplasmic ERK activation is an independent adverse prognostic factor in primary cutaneous melanoma. AB - The aim of this study was to estimate the impact on survival of NRAS and BRAF mutations and activation of Akt and extracellular signal-regulated kinase (ERK) in primary melanomas. A cohort of 57 primary cutaneous T1-2 melanoma tumors was analyzed. Mutation frequency for both genes was 61% (NRAS 26% and BRAF 39%). In a univariate analysis, shorter overall survival was associated with the presence of ulceration (P=0.001) and BRAF exon 15 mutations (P=0.005) as well as the absence of nuclear activation of Akt (P=0.022) and of cytoplasmic activation of ERK (P=0.003). Unexpectedly, ulceration was a significant adverse prognostic factor only in melanomas with BRAF mutations, whereas there was no effect of ulceration on overall survival in tumors with wild-type BRAF. A multivariate analysis showed that significant independent adverse survival prognostic markers were absence of cytoplasmic activation of ERK (P=0.007) and ulceration (P=0.008), whereas BRAF exon 15 mutation status showed a nonsignificant trend (P=0.066). The absence of cytoplasmic ERK activation in poor prognosis T1-2 melanomas may be associated with activation of some other uncharacterized pathway leading to tumor progression and adverse outcome. Immunohistochemical analysis of cytoplasmic phosphorylated ERK could be used as a prognostic marker in primary melanomas if confirmed in another data set. PMID- 18509362 TI - Inactivation of the vitamin D receptor enhances susceptibility of murine skin to UV-induced tumorigenesis. AB - 1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) is the biologically active ligand for the vitamin D receptor (VDR). VDR(-/-) mice have a hair follicle-cycling defect resulting in alopecia. However, mice lacking 25-hydroxyvitamin D(3) 1alpha hydroxylase (CYP27B1(-/-)), and having no circulating 1,25(OH)(2)D(3), have normal follicular function. These mouse models indicate that VDR functions independently of 1,25(OH)(2)D(3) in regulating hair-follicle cycling. Here, we show that VDR(-/-) mice rapidly develop chemically induced skin tumors, whereas CYP27B1(-/-) and wild-type mice do not, indicating that VDR, and not the 1,25(OH)(2)D(3) ligand, is essential for protection against skin tumorigenesis. Because the majority of human skin cancer results from exposure to UV, the susceptibility of VDR(-/-) mice to this carcinogen was also evaluated. VDR(-/-) mice developed UV-induced tumors more rapidly and with greater penetrance than did VDR(+/+) mice. p53 protein levels were upregulated at similar rates in UV treated keratinocytes of VDR(-/-) and VDR(+/+) mice. However, rates of thymine dimer repair and UV-induced apoptosis were significantly lower in VDR(-/-) epidermis compared with the wild type epidermis. UV-induced epidermal thickening was also attenuated in VDR(-/-) skin, indicating that VDR plays a critical role in the repair and removal of severely damaged keratinocytes and adaptation of the skin to chronic UV exposure. PMID- 18509364 TI - Special issue II: Clinical Sandor Ferenczi Conference in Baden-Baden. PMID- 18509363 TI - Primary cutaneous T-cell lymphomas do not show specific NAV3 gene deletion or translocation. AB - The mapping of a balanced t(12;18)(q21;q21.2) translocation in a Sezary syndrome (SS) case led Karenko et al. to identify NAV3 gene (12q21-22) deletion by interphase fluorescence in situ hybridization (FISH) in 15/21 patients with mycosis fungoides (MF) or SS. To determine whether the NAV3 deletion is the result of a specific gene breakpoint, we used FISH with dual-color split or break apart bacterial artificial chromosome (BAC) probes covering the NAV3 locus. A total of 31 samples (18 skin, 11 blood, 1 lymph node, and 1 spleen) from 24 patients with advanced MF/SS (18 with large-cell transformation) were studied. Chromosome 12 imbalances were analyzed by comparative genomic hybridization (CGH) array with a 3K BAC probes in 24 samples from 22 patients. Both normal FISH and CGH array patterns were observed in 22 samples from 18 patients. In 6 patients, abnormal patterns were observed with an abnormal number of chromosome 12 set in 5 of them. Chromosome 12 structural abnormalities were seen in four of these six patients. An imbalanced FISH pattern between NAV3 and pericentromeric control probes was seen in three patients in accordance with CGH array data (one with a pericentromeric deletion and two with a large 12q deletion including NAV3). No NAV3 specific breakpoint or partial deletion was detected. PMID- 18509365 TI - Freud, his illness, and ourselves. AB - The history of Freud's illness shows that he tried to avoid confrontation with it, and to treat it as unimportant. In his personal letters, the ill body remains outside-as another person, "Konrad," not he himself-and it is not taken into account. Particularly in Freud's correspondence with Ferenczi, we realize to what extent certain phenomena, especially depressive ones, he considered somatic, with a tendency to dismiss them, and this despite important occasional insights, such as about the role played by hate in psychosomatic illnesses. In the post-Freudian development, these topics have been more and more integrated in the dialogue, in the discourse between the analyst and the analysand. PMID- 18509366 TI - Inventing Freud. AB - Written to commemorate the 150th anniversary of Freud's birth, this paper construes Nina Coltart's statement that "if Freud did not exist it would be necessary to invent him," with its implicit comparison of Freud to God, to refer to (a) the things that Freud taught that are incontrovertibly true; (b) the unavoidable subjectivity in all judgments of Freud; and (c) the resemblances between psychoanalysis and religion. This last comparison is likewise seen to have both positive and negative aspects. Freud's ideas have inspired many people, yet he unscientifically arrogated sovereign authority over psychoanalysis. Freud's admirers are reminded of his extreme difficulty in admitting he was wrong and changing his mind when he should have known better, while his detractors are encouraged to consider the evidence supporting many of Freud's core tenets and to recognize that his discovery of psychoanalysis is indeed one of the supreme achievements in human history. PMID- 18509367 TI - Ferenczi's collaboration with rank: on paradigm shift and the origins of complementarity in psychoanalysis. AB - The publication in 1923 of Sigmund Freud's The Ego and the Id is generally recognized as a watershed event in the history of psychoanalysis. The following year Ferenczi and Rank jointly published a book on psychoanalytic technique, The Development of Psychoanalysis (Ger: Entwicklungsziele der Psychoanalyse), concurrently with the independent publication of Ferenczi's Thalassa: A Theory of Genitality (Ger: Versuch einer Genitaltheorie) and Rank's The Trauma of Birth, the latter of which sparked a storm of controversy within the psychoanalytic community. The ensuing debate over technical and theoretical issues raised in these works signaled the emergence of two rival paradigms (following the model described by Thomas Kuhn) in psychoanalytic technique, the one based on ego psychology and drive-dynamics, the other on preoedipal ego development and object relations, which, when viewed from a present-day vantage point, may be considered complementary. The current crisis in psychoanalysis represents a continuation of this not yet completed paradigm shift. PMID- 18509368 TI - Definition, foundation and meaning of illness: locating Georg Groddeck in the history of medicine. AB - The paper reviews and discusses Groddeck's conception of illness. I first argue that Groddeck was a late Romanticist as much as he was a "wild" psychoanalyst. Then I use Groddeck's scattered formulations regarding definition, foundation, and meaning of illness in order to articulate them in the form of more explicit scientific statements. Finally, I suggest that Groddeck's theory of illness is fundamentally different from current medical conceptions, which, nevertheless, does not make our dialogue with him either less useful or indeed less necessary. It is through an investigation of Groddeck's therapeutic attitude and the spirit of his work that psychoanalysis and medicine can build foundations of bold, innovative, and healing future. PMID- 18509369 TI - Psychoanalysis, a bridge between attachment research and neurobiology. AB - Attachment theory and neurobiology are at the forefront of scientific research, particularly in the area of child psychiatry. Several authors have encountered a surprising isomorphism between findings in these areas and concepts central in psychoanalysis. The author postulates that attachment theory is an outcome of the history of the transformation of the old concept of libido now applied interpersonally. The author also postulates that the neurobiology of attachment mediates but does not substitute psychoanalysis for the final understanding of the human bond. Findings in both fields, attachment theory and psychobiology, often compellingly lead to psychoanalytic concepts; on the other hand, the neurobiology of the psyche does not make full sense without the wealth of research conducted by psychoanalysts during the 20th century. The concept of seduction in particular, in its broad sense, is akin to mirroring and attachment and a useful tool in this regard. Although some cling to the idea of the purity of psychoanalysis and see these kin areas as a threat to its integrity, the author believes that psychoanalysis, serving as a bridge between them, can only be enriched by their empirical and experimental outcomes. The American Journal of Psychoanalysis (2008) 68, 148-155. doi:10.1057/ajp.2008.4. PMID- 18509370 TI - The man who could not cry and the psychoanalyst who could: mutual healing in the maternal transference/countertransference. AB - This is a clinical/theoretical study of mutual healing in the maternal transference/countertransference. Therapist and patient began their work when each was in extreme mourning. I detail sensory, affective transactions between them that proved transformative. I deem these basic to the healing that was unusually rapid. It broke through the patient's dissociation, and the therapist's sorrow. I note that the treatment modalities were similar to those essential to mother/infant bonding. I connect the treatment's transformative effects with those discovered through infant research and studies with the functioning magnetic resonance imagery test (fMRI). All three of these modes of interpersonal study-psychotherapy, infant, and fMRI research-exhibit mind/body imprinting of the mother/infant deeply connected experience. PMID- 18509371 TI - Fear of the mind. The annihilating power of the gaze. AB - The concern for the annihilating power of the gaze is not part of the Freudian discourse, but represents one of the most valuable contributions of phenomenology to psychoanalysis. When clear boundaries between the self and the others are not yet established, the gaze is experienced as a disembodied force that radiates from the eyes and can dangerously penetrate into the mind. In this regard, the body or parts of it can be used as a shelter. If the external body is not sufficiently cathected, its sheltering function is also decreased, to the point that the body is experienced as transparent, and the most intimate feelings and thoughts become dangerously available to the others. In primitive societies this situation is experienced as the danger of losing the soul. The unconscious fantasy of obstructing the sight can be used to neutralize the annihilating power of the gaze by introducing an artificial barrier between the minds. In dreams and in other expressions of the unconscious, the black color might hint at such an artificial barrier. What is then blackened are moments of the meeting of the mind that cannot be introjected. Blind spots in the perception of the mind of the other as well as in the perception of the self are a specific consequence of this kind of defense. PMID- 18509372 TI - Allergic to people: building bridges in a ripped psychic-soma. AB - An exploration of the use mind/body metaphors in a woman whose physical, environmental and psychoneurotic trauma culminated in an irreversible colostomy. She lived in a world of concrete symbols, her primary process damaged such that she could not create generative symbols to process her trauma. She regressed to a state of infantile megalomania, recoiling from the external reality of subjective others. Her introjective disorder mirrored her digestive disorder as she could absorb neither good objects nor good nutrients. The analytic situation has been an auxiliary fecal container and we work to bridge her mind body split with mind/body metaphors. As she reclaims lost development mastery, she displays a symbolized sphincter. As her capacity to form symbols grows, she rages and mourns for the loss of her fantasized ideal parents and her ideal body. PMID- 18509373 TI - A conversation about psychoanalysis, literature and anthropology between Anthony Molino, and Benjamin Kilborne, Date: May 18, 2007. PMID- 18509378 TI - Syncytia formation affects the yield and cytotoxicity of an adenovirus expressing a fusogenic glycoprotein at a late stage of replication. AB - Fusogenic membrane glycoproteins (FMGs) may enhance the cytotoxicity of conditionally replicative adenoviruses. However, expression at early stages of infection impairs virus replication. We have inserted the hyperfusogenic form of the gibbon ape leukemia virus (GALV) envelope glycoprotein as a new splice unit of the major late promoter (MLP) to generate a replication-competent adenovirus expressing this protein. At high multiplicity of infection (MOI), this virus replicated efficiently forming clumps of fused cells and showing a faster release. In contrast, at low MOI, infected cells formed syncytia where only one nucleus contained virus DNA, decreasing total virus production but increasing cytotoxicity. PMID- 18509379 TI - Targeted inhibition of platelet-derived growth factor receptor-beta subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats. AB - The activation of hepatic stellate cells (HSCs) is the key event of the pathogenesis of hepatic fibrosis. Platelet-derived growth factor (PDGF) is the most potent mitogen for HSCs, and PDGF receptor-beta subunit (PDGFR-beta) is required for the proliferation of HSCs induced by PDGF. In this study, a high gene-silencing-efficacy PDGFR-beta small interference RNA (siRNA) was synthesized that could suppress the PDGFR-beta expression and inhibit the activation and proliferation but could not induce the apoptosis of HSCs in vitro. To avoid the side effect of nonspecific interference of PDGFR-beta, we constructed an HSCs specific short hairpin RNA (shRNA) expression plasmid in which PDGFR-beta shRNA was driven by a glial fibrillary acidic protein (GFAP) promoter. The double staining immunofluorescence examination indicated that GFAP promoter could target the transgene expression into HSCs in carbon tetrachloride induced acute injured rat's liver and bile duct ligation (BDL)-induced chronic injured rat's liver. Furthermore, HSCs-specific PDGFR-beta shRNA could relieve liver injury and hepatic fibrosis in the rat's model induced by BDL. This study demonstrates that PDGFR-beta siRNA may be presented as an antifibrogenic agent. The application of HSCs-specific RNA interference induced by the GFAP promoter might supply a new powerful tool for cell-specific gene therapy of hepatic fibrogenesis. PMID- 18509380 TI - Cell entry targeting restricts biodistribution of replication-competent retroviruses to tumour tissue. AB - Virotherapy is currently being developed for many different types of viruses including replication-competent murine leukaemia virus (MLV) as a novel tool in cancer therapy. However, there is the risk of insertional mutagenesis associated with this virus, making careful preclinical studies necessary before its first application in man. We have previously generated conditionally replication competent MLV variants that require activation by tumour-associated proteases to become infectious. Here we analysed in a comparative study the spreading of non targeted and of such tumour-targeted MLV variants to tumour and extratumoural organs in immunodeficient mice. Both virus types were able to efficiently infect tumour cells after systemic administration. The non-targeted virus, however, also infected extratumoural organs like bone marrow, spleen and liver efficiently. In contrast, the targeted viruses revealed in a quantitative analysis of virus spreading an up to 500-fold more selective infection of tumour tissue than the non-targeted virus. The data raise serious doubts about a safe clinical use of non-targeted MLV. Engineering the virus to become activatable by tumour associated proteases can significantly improve the safety of MLV. PMID- 18509381 TI - Isolation of myogenic progenitor populations from Pax7-deficient skeletal muscle based on adhesion characteristics. AB - In an attempt to determine whether muscle-derived stem cells are distinct from satellite cells, we investigated whether muscle-derived stem cells could be isolated from the skeletal muscle of Pax7-deficient mice, which have been shown to be devoid of or to contain only a minimal number of satellite cells. Utilizing a technique that separates cells based on their adhesion characteristics (the preplate technique), several distinct populations of muscle-derived cells were isolated. In these mice, the Pax7 gene was knocked out with the insertion of the LacZ gene. One population was both rapidly adhering, LacZ-positive, and displayed a high myogenic index, but was rapidly lost to terminal differentiation when continuously replated. A second population, which persisted over 50 passages, was LacZ-negative and displayed a low myogenic index. Although Pax3 may have acted as a compensatory mechanism for the myogenic commitment of the LacZ-positive cells, the LacZ-negative cells, despite expressing Pax3, required Pax7 transduction to restore their myogenic capacity. We believe that these two populations of myogenic progenitor cells, each endowed with different adhesion characteristics, may help explain the discrepancy in the literature concerning the presence of myogenic cells found in Pax7-deficient mice. PMID- 18509382 TI - Function and evolution of the psbA gene family in marine Synechococcus: Synechococcus sp. WH7803 as a case study. AB - In cyanobacteria, the D1 protein of photosystem II (PSII) is encoded by the psbA multigene family. In most freshwater strains, a D1:1 isoform of this protein is exchanged for a D1:2 isoform in response to various stresses, thereby altering PSII photochemistry. To investigate PSII responses to stress in marine Synechococcus, we acclimated cultures of the WH7803 strain to different growth irradiances and then exposed them to high light (HL) or ultraviolet (UV) radiation. Measurement of PSII quantum yield and quantitation of the D1 protein pool showed that HL-acclimated cells were more resistant to UV light than were low light- (LL) or medium light- (ML) acclimated cells. Both UV and HL induced the expression of psbA genes encoding D1:2 and the repression of the psbA gene encoding D1:1. Although three psbA genes encode identical D1:2 isoforms in Synechococcus sp. WH7803, only one was strongly stress responsive in our treatment conditions. Examination of 11 marine Synechococcus genomic sequences identified up to six psbA copies per genome, with always a single gene encoding D1:1. In phylogenetic analyses, marine Synechococcus genes encoding D1:1 clustered together, while the genes encoding D1:2 grouped by genome into subclusters. Moreover, examination of the genomic environment of psbA genes suggests that the D1:2 genes are hotspots for DNA recombination. Collectively, our observations suggest that while all psbA genes follow a concerted evolution within each genome, D1:2 coding genes are subject to intragenome homogenization most probably mediated by gene conversion. PMID- 18509386 TI - Animal tests inescapable. PMID- 18509387 TI - Who are we? PMID- 18509388 TI - Linking up development. PMID- 18509400 TI - Venerable institute gets a refit. PMID- 18509401 TI - Climate anomaly is an artefact. PMID- 18509402 TI - Genetic testing for everyone. PMID- 18509403 TI - France's research agency splits up. PMID- 18509405 TI - The oldest pregnant mum. PMID- 18509412 TI - Microbiology: the inside story. PMID- 18509413 TI - Microbiology: straight from the gut. PMID- 18509414 TI - Marine microbiology: origins of death. PMID- 18509415 TI - Analyses support theory of stochastic regulation of fisheries. PMID- 18509416 TI - A prime problem that even quantum computing can't solve. PMID- 18509417 TI - Costa Rica's biotech project still on track for end of year. PMID- 18509418 TI - Increasing use of stimulants warns of potential abuse. PMID- 18509419 TI - Time to break the silence around virtual-water imports. PMID- 18509420 TI - Ancient asexuals: darwinulids not exposed. PMID- 18509421 TI - If you don't need change, maybe you don't need sex. PMID- 18509422 TI - Retiring retirement. PMID- 18509429 TI - Science & music: the neural roots of music. PMID- 18509430 TI - Climate change: hot questions of temperature bias. PMID- 18509431 TI - Immunology: soothing intestinal sugars. PMID- 18509432 TI - Cancer: whispering sweet somethings. PMID- 18509433 TI - Evolutionary biology: sex ratios writ small. PMID- 18509434 TI - Mathematical physics: packings close and loose. PMID- 18509435 TI - Sex ratio adjustment and kin discrimination in malaria parasites. AB - Malaria parasites and related Apicomplexans are the causative agents of the some of the most serious infectious diseases of humans, companion animals, livestock and wildlife. These parasites must undergo sexual reproduction to transmit from vertebrate hosts to vectors, and their sex ratios are consistently female-biased. Sex allocation theory, a cornerstone of evolutionary biology, is remarkably successful at explaining female-biased sex ratios in multicellular taxa, but has proved controversial when applied to malaria parasites. Here we show that, as predicted by theory, sex ratio is an important fitness-determining trait and Plasmodium chabaudi parasites adjust their sex allocation in response to the presence of unrelated conspecifics. This suggests that P. chabaudi parasites use kin discrimination to evaluate the genetic diversity of their infections, and they adjust their behaviour in response to environmental cues. Malaria parasites provide a novel way to test evolutionary theory, and support the generality and power of a darwinian approach. PMID- 18509436 TI - A microbial symbiosis factor prevents intestinal inflammatory disease. AB - Humans are colonized by multitudes of commensal organisms representing members of five of the six kingdoms of life; however, our gastrointestinal tract provides residence to both beneficial and potentially pathogenic microorganisms. Imbalances in the composition of the bacterial microbiota, known as dysbiosis, are postulated to be a major factor in human disorders such as inflammatory bowel disease. We report here that the prominent human symbiont Bacteroides fragilis protects animals from experimental colitis induced by Helicobacter hepaticus, a commensal bacterium with pathogenic potential. This beneficial activity requires a single microbial molecule (polysaccharide A, PSA). In animals harbouring B. fragilis not expressing PSA, H. hepaticus colonization leads to disease and pro inflammatory cytokine production in colonic tissues. Purified PSA administered to animals is required to suppress pro-inflammatory interleukin-17 production by intestinal immune cells and also inhibits in vitro reactions in cell cultures. Furthermore, PSA protects from inflammatory disease through a functional requirement for interleukin-10-producing CD4+ T cells. These results show that molecules of the bacterial microbiota can mediate the critical balance between health and disease. Harnessing the immunomodulatory capacity of symbiosis factors such as PSA might potentially provide therapeutics for human inflammatory disorders on the basis of entirely novel biological principles. PMID- 18509437 TI - An infrared ring around the magnetar SGR 1900+14. AB - Magnetars are a special class of slowly rotating (period approximately 5-12 s) neutron stars with extremely strong magnetic fields (>10(14 )G)--at least an order of magnitude larger than those of the 'normal' radio pulsars. The potential evolutionary links and differences between these two types of object are still unknown; recent studies, however, have provided circumstantial evidence connecting magnetars with very massive progenitor stars. Here we report the discovery of an infrared elliptical ring or shell surrounding the magnetar SGR 1900+14. The appearance and energetics of the ring are difficult to interpret within the framework of the progenitor's stellar mass loss or the subsequent evolution of the supernova remnant. We suggest instead that a dust-free cavity was produced in the magnetar environment by the giant flare emitted by the source in August 1998. Considering the total energy released in the flare, the theoretical dust-destruction radius matches well with the observed dimensions of the ring. We conclude that SGR 1900+14 is unambiguously associated with a cluster of massive stars, thereby solidifying the link between magnetars and massive stars. PMID- 18509438 TI - A phase diagram for jammed matter. AB - The problem of finding the most efficient way to pack spheres has a long history, dating back to the crystalline arrays conjectured by Kepler and the random geometries explored by Bernal. Apart from its mathematical interest, the problem has practical relevance in a wide range of fields, from granular processing to fruit packing. There are currently numerous experiments showing that the loosest way to pack spheres (random loose packing) gives a density of approximately 55 per cent. On the other hand, the most compact way to pack spheres (random close packing) results in a maximum density of approximately 64 per cent. Although these values seem to be robust, there is as yet no physical interpretation for them. Here we present a statistical description of jammed states in which random close packing can be interpreted as the ground state of the ensemble of jammed matter. Our approach demonstrates that random packings of hard spheres in three dimensions cannot exceed a density limit of approximately 63.4 per cent. We construct a phase diagram that provides a unified view of the hard-sphere packing problem and illuminates various data, including the random-loose-packed state. PMID- 18509439 TI - Quantum phase transition in a single-molecule quantum dot. AB - Quantum criticality is the intriguing possibility offered by the laws of quantum mechanics when the wave function of a many-particle physical system is forced to evolve continuously between two distinct, competing ground states. This phenomenon, often related to a zero-temperature magnetic phase transition, is believed to govern many of the fascinating properties of strongly correlated systems such as heavy-fermion compounds or high-temperature superconductors. In contrast to bulk materials with very complex electronic structures, artificial nanoscale devices could offer a new and simpler means of understanding quantum phase transitions. Here we demonstrate this possibility in a single-molecule quantum dot, where a gate voltage induces a crossing of two different types of electron spin state (singlet and triplet) at zero magnetic field. The quantum dot is operated in the Kondo regime, where the electron spin on the quantum dot is partially screened by metallic electrodes. This strong electronic coupling between the quantum dot and the metallic contacts provides the strong electron correlations necessary to observe quantum critical behaviour. The quantum magnetic phase transition between two different Kondo regimes is achieved by tuning gate voltages and is fundamentally different from previously observed Kondo transitions in semiconductor and nanotube quantum dots. Our work may offer new directions in terms of control and tunability for molecular spintronics. PMID- 18509440 TI - Anatase TiO2 single crystals with a large percentage of reactive facets. AB - Owing to their scientific and technological importance, inorganic single crystals with highly reactive surfaces have long been studied. Unfortunately, surfaces with high reactivity usually diminish rapidly during the crystal growth process as a result of the minimization of surface energy. A typical example is titanium dioxide (TiO2), which has promising energy and environmental applications. Most available anatase TiO(2) crystals are dominated by the thermodynamically stable {101} facets (more than 94 per cent, according to the Wulff construction), rather than the much more reactive {001} facets. Here we demonstrate that for fluorine terminated surfaces this relative stability is reversed: {001} is energetically preferable to {101}. We explored this effect systematically for a range of non metallic adsorbate atoms by first-principle quantum chemical calculations. On the basis of theoretical predictions, we have synthesized uniform anatase TiO(2) single crystals with a high percentage (47 per cent) of {001} facets using hydrofluoric acid as a morphology controlling agent. Moreover, the fluorated surface of anatase single crystals can easily be cleaned using heat treatment to render a fluorine-free surface without altering the crystal structure and morphology. PMID- 18509441 TI - Snowball Earth termination by destabilization of equatorial permafrost methane clathrate. AB - The start of the Ediacaran period is defined by one of the most severe climate change events recorded in Earth history--the recovery from the Marinoan 'snowball' ice age, approximately 635 Myr ago (ref. 1). Marinoan glacial-marine deposits occur at equatorial palaeolatitudes, and are sharply overlain by a thin interval of carbonate that preserves marine carbon and sulphur isotopic excursions of about -5 and +15 parts per thousand, respectively; these deposits are thought to record widespread oceanic carbonate precipitation during postglacial sea level rise. This abrupt transition records a climate system in profound disequilibrium and contrasts sharply with the cyclical stratigraphic signal imparted by the balanced feedbacks modulating Phanerozoic deglaciation. Hypotheses accounting for the abruptness of deglaciation include ice albedo feedback, deep-ocean out-gassing during post-glacial oceanic overturn or methane hydrate destabilization. Here we report the broadest range of oxygen isotope values yet measured in marine sediments (-25 per thousand to +12 per thousand) in methane seeps in Marinoan deglacial sediments underlying the cap carbonate. This range of values is likely to be the result of mixing between ice-sheet-derived meteoric waters and clathrate-derived fluids during the flushing and destabilization of a clathrate field by glacial meltwater. The equatorial palaeolatitude implies a highly volatile shelf permafrost pool that is an order of magnitude larger than that of the present day. A pool of this size could have provided a massive biogeochemical feedback capable of triggering deglaciation and accounting for the global postglacial marine carbon and sulphur isotopic excursions, abrupt unidirectional warming, cap carbonate deposition, and a marine oxygen crisis. Our findings suggest that methane released from low-latitude permafrost clathrates therefore acted as a trigger and/or strong positive feedback for deglaciation and warming. Methane hydrate destabilization is increasingly suspected as an important positive feedback to climate change that coincides with critical boundaries in the geological record and may represent one particularly important mechanism active during conditions of strong climate forcing. PMID- 18509442 TI - A large discontinuity in the mid-twentieth century in observed global-mean surface temperature. AB - Data sets used to monitor the Earth's climate indicate that the surface of the Earth warmed from approximately 1910 to 1940, cooled slightly from approximately 1940 to 1970, and then warmed markedly from approximately 1970 onward. The weak cooling apparent in the middle part of the century has been interpreted in the context of a variety of physical factors, such as atmosphere-ocean interactions and anthropogenic emissions of sulphate aerosols. Here we call attention to a previously overlooked discontinuity in the record at 1945, which is a prominent feature of the cooling trend in the mid-twentieth century. The discontinuity is evident in published versions of the global-mean temperature time series, but stands out more clearly after the data are filtered for the effects of internal climate variability. We argue that the abrupt temperature drop of approximately 0.3 degrees C in 1945 is the apparent result of uncorrected instrumental biases in the sea surface temperature record. Corrections for the discontinuity are expected to alter the character of mid-twentieth century temperature variability but not estimates of the century-long trend in global-mean temperatures. PMID- 18509443 TI - Live birth in the Devonian period. AB - The extinct placoderm fishes were the dominant group of vertebrates throughout the Middle Palaeozoic era, yet controversy about their relationships within the gnathostomes (jawed vertebrates) is partly due to different interpretations of their reproductive biology. Here we document the oldest record of a live-bearing vertebrate in a new ptyctodontid placoderm, Materpiscis attenboroughi gen. et sp. nov., from the Late Devonian Gogo Formation of Australia (approximately 380 million years ago). The new specimen, remarkably preserved in three dimensions, contains a single, intra-uterine embryo connected by a permineralized umbilical cord. An amorphous crystalline mass near the umbilical cord possibly represents the recrystallized yolk sac. Another ptyctodont from the Gogo Formation, Austroptyctodus gardineri, also shows three small embryos inside it in the same position. Ptyctodontids have already provided the oldest definite evidence for vertebrate copulation, and the new specimens confirm that some placoderms had a remarkably advanced reproductive biology, comparable to that of some modern sharks and rays. The new discovery points to internal fertilization and viviparity in vertebrates as originating earliest within placoderms. PMID- 18509446 TI - Metagenomics: exploring unseen communities. PMID- 18509444 TI - Abundance and diversity of microbial life in ocean crust. AB - Oceanic lithosphere exposed at the sea floor undergoes seawater-rock alteration reactions involving the oxidation and hydration of glassy basalt. Basalt alteration reactions are theoretically capable of supplying sufficient energy for chemolithoautotrophic growth. Such reactions have been shown to generate microbial biomass in the laboratory, but field-based support for the existence of microbes that are supported by basalt alteration is lacking. Here, using quantitative polymerase chain reaction, in situ hybridization and microscopy, we demonstrate that prokaryotic cell abundances on seafloor-exposed basalts are 3-4 orders of magnitude greater than in overlying deep sea water. Phylogenetic analyses of basaltic lavas from the East Pacific Rise (9 degrees N) and around Hawaii reveal that the basalt-hosted biosphere harbours high bacterial community richness and that community membership is shared between these sites. We hypothesize that alteration reactions fuel chemolithoautotrophic microorganisms, which constitute a trophic base of the basalt habitat, with important implications for deep-sea carbon cycling and chemical exchange between basalt and sea water. PMID- 18509450 TI - Th1 disabled function in response to TLR4 stimulation of monocyte-derived DC from patients chronically-infected by hepatitis C virus. AB - BACKGROUND: Lack of protective antibodies and inefficient cytotoxic responses are characteristics of chronic hepatitis C infection. A defect in dendritic cell (DC) function has thus been suspected, but this remains a controversial issue. METHODS AND FINDINGS: Here we show that monocyte-derived DC (MoDC) from chronically infected patients can mature in response to TLR1/2, TLR2/6 or TLR3 ligands. In contrast, when stimulated with the TLR4 ligand LPS, MoDC from patients show a profound defect in inducing IFNgamma secretion by allogeneic T cells. This defect is not due to defective phenotypic maturation or to the presence of HCV-RNA in DC or monocytes but is correlated to reduced IL-12 secretion by DC. Restoration of DC ability to stimulate IFNgamma secretion can be obtained by blocking MEK activation in DC, indicating that MEK/ERK pathway is involved in the Th1 defect of MoDC. Monocytes from HCV patients present increased spontaneous secretion of cytokines and chemokines, especially MIP-1beta. Addition of MIP-1beta on healthy monocytes during differentiation results in DC that have Th1 defect characteristic of MoDC from HCV patients, suggesting that MIP-1beta secretion by HCV monocytes participates in the Th1 defect of DC. CONCLUSIONS: Our data indicate that monocytes from HCV patients are activated in vivo. This interferes with their differentiation into DC, leading to deficient TLR4 signaling in these cells that are enable to induce a Th1 response. This specific defect is linked to the activation of the MEK/ERK pathway. PMID- 18509451 TI - Orientation sensitivity at different stages of object processing: evidence from repetition priming and naming. AB - BACKGROUND: An ongoing debate in the object recognition literature centers on whether the shape representations used in recognition are coded in an orientation dependent or orientation-invariant manner. In this study, we asked whether the nature of the object representation (orientation-dependent vs orientation invariant) depends on the information-processing stages tapped by the task. METHODOLOGY/FINDINGS: We employed a repetition priming paradigm in which briefly presented masked objects (primes) were followed by an upright target object which had to be named as rapidly as possible. The primes were presented for variable durations (ranging from 16 to 350 ms) and in various image-plane orientations (from 0 degrees to 180 degrees, in 30 degrees steps). Significant priming was obtained for prime durations above 70 ms, but not for prime durations of 16 ms and 47 ms, and did not vary as a function of prime orientation. In contrast, naming the same objects that served as primes resulted in orientation-dependent reaction time costs. CONCLUSIONS/SIGNIFICANCE: These results suggest that initial processing of object identity is mediated by orientation-independent information and that orientation costs in performance arise when objects are consolidated in visual short-term memory in order to be reported. PMID- 18509452 TI - Mouse apolipoprotein B editing complex 3 (APOBEC3) is expressed in germ cells and interacts with dead-end (DND1). AB - BACKGROUND: The dead-end (Dnd1) gene is essential for maintaining the viability of germ cells. Inactivation of Dnd1 results in sterility and testicular tumors. The Dnd1 encoded protein, DND1, is able to bind to the 3'-untranslated region (UTR) of messenger RNAs (mRNAs) to displace micro-RNA (miRNA) interaction with mRNA. Thus, one function of DND1 is to prevent miRNA mediated repression of mRNA. We report that DND1 interacts specifically with APOBEC3. APOBEC3 is a multi functional protein. It inhibits retroviral replication. In addition, recent studies show that APOBEC3 interacts with cellular RNA-binding proteins and to mRNA to inhibit miRNA-mediated repression of mRNA. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that DND1 specifically interacts with another cellular protein, APOBEC3. We present our data which shows that DND1 co-immunoprecipitates APOBEC3 from mammalian cells and also endogenous APOBEC3 from mouse gonads. Whether the two proteins interact directly remains to be elucidated. We show that both DND1 and APOBEC3 are expressed in germ cells and in the early gonads of mouse embryo. Expression of fluorescently-tagged DND1 and APOBEC3 indicate they localize to the cytoplasm and when DND1 and APOBEC3 are expressed together in cells, they sequester near peri-nuclear sites. CONCLUSIONS/SIGNIFICANCE: The 3' UTR of mRNAs generally encode multiple miRNA binding sites as well as binding sites for a variety of RNA binding proteins. In light of our findings of DND1 APOBEC3 interaction and taking into consideration reports that DND1 and APOBEC3 bind to mRNA to inhibit miRNA mediated repression, our studies implicate a possible role of DND1-APOBEC3 interaction in modulating miRNA-mediated mRNA repression. The interaction of DND1 and APOBEC3 could be one mechanism for maintaining viability of germ cells and for preventing germ cell tumor development. PMID- 18509453 TI - Microbiological implications of periurban agriculture and water reuse in Mexico City. AB - BACKGROUND: Recycled treated or untreated wastewater represents an important health challenge in developing countries due to potential water related microbiological exposure. Our aim was to assess water quality and health implications in a Mexico City periurban agricultural area. METHODOLOGY/PRINCIPAL FINDINGS: A longitudinal study in the Xochimilco wetland area was conducted, and 42 sites were randomly selected from 211, including irrigation water canals and effluents of treatment plants. Sample collection took place during rainy and dry seasons (2000-2001). Microbiological parameters (total coliforms, fecal coliforms, streptococci/enterococci, and bacteria other than Vibrio grown on TCBS), Helicobacter pylori, and physicochemical parameters including trihalomethanes (THM) were determined. Fecal coliforms and fecal streptococci are appropriate indicators of human or animal fecal contamination. Fecal coliform counts surpass Mexican and World Health Organization irrigation water guidelines. Identified microorganisms associated with various pathologies in humans and domestic animals comprise Escherichia coli, Klebsiella spp., Salmonella spp., Enterobacter spp., Enterococcus spp., and Pseudomonas spp; H. pylori was also present in the water. An environmental characteristic of the canal system showed high Total Organic Carbon content and relatively low dissolved oxygen concentration; residual chlorine as a disinfection control is not efficient, but THMs do not represent a problem. During the rainy season, temperature and conductivity were higher; in contrast, pH, dissolved oxygen, ammonia, and residual chlorine were lower. This is related with the continuous load of feces from human and animal sources, and to the aquatic systems, which vary seasonally and exhibit evidence of lower water quality in effluents from treatment plants. CONCLUSIONS/SIGNIFICANCE: There is a need for improvement of wastewater treatment systems, as well as more efficient monitoring, regulation, and enforcement procedures for wastewater disposal into bodies of water. PMID- 18509454 TI - Variations of plasmid content in Rickettsia felis. AB - BACKGROUND: Since its first detection, characterization of R. felis has been a matter of debate, mostly due to the contamination of an initial R. felis culture by R. typhi. However, the first stable culture of R. felis allowed its precise phenotypic and genotypic characterization, and demonstrated that this species belonged to the spotted fever group rickettsiae. Later, its genome sequence revealed the presence of two forms of the same plasmid, physically confirmed by biological data. In a recent article, Gillespie et al. (PLoS One. 2007;2(3):e266.) used a bioinformatic approach to refute the presence of the second plasmid form, and proposed the creation of a specific phylogenetic group for R. felis. METHODOLOGY/PRINCIPAL FINDINGS: In the present report, we, and five independent international laboratories confirmed unambiguously by PCR the presence of two plasmid forms in R. felis strain URRWXCal(2) (T), but observed that the plasmid content of this species, from none to 2 plasmid forms, may depend on the culture passage history of the studied strain. We also demonstrated that R. felis does not cultivate in Vero cells at 37 degrees C but generates plaques at 30 degrees C. Finally, using a phylogenetic study based on 667 concatenated core genes, we demonstrated the position of R. felis within the spotted fever group. SIGNIFICANCE: We demonstrated that R. felis, which unambiguously belongs to the spotted fever group rickettsiae, may contain up to two plasmid forms but this plasmid content is unstable. PMID- 18509455 TI - Why are clinicians not embracing the results from pivotal clinical trials in severe sepsis? A bayesian analysis. AB - BACKGROUND: Five pivotal clinical trials (Intensive Insulin Therapy; Recombinant Human Activated Protein C [rhAPC]; Low-Tidal Volume; Low-Dose Steroid; Early Goal Directed Therapy [EGDT]) demonstrated mortality reduction in patients with severe sepsis and expert guidelines have recommended them to clinical practice. Yet, the adoption of these therapies remains low among clinicians. OBJECTIVES: We selected these five trials and asked: Question 1--What is the current probability that the new therapy is not better than the standard of care in my patient with severe sepsis? Question 2--What is the current probability of reducing the relative risk of death (RRR) of my patient with severe sepsis by meaningful clinical thresholds (RRR >15%; >20%; >25%)? METHODS: Bayesian methodologies were applied to this study. Odds ratio (OR) was considered for Question 1, and RRR was used for Question 2. We constructed prior distributions (enthusiastic; mild, moderate, and severe skeptic) based on various effective sample sizes of other relevant clinical trials (unfavorable evidence). Posterior distributions were calculated by combining the prior distributions and the data from pivotal trials (favorable evidence). MAIN FINDINGS: Answer 1--The analysis based on mild skeptic prior shows beneficial results with the Intensive Insulin, rhAPC, and Low-Tidal Volume trials, but not with the Low-Dose Steroid and EGDT trials. All trials' results become unacceptable by the analyses using moderate or severe skeptic priors. Answer 2--If we aim for a RRR>15%, the mild skeptic analysis shows that the current probability of reducing death by this clinical threshold is 88% for the Intensive Insulin, 62-65% for the Low-Tidal Volume, rhAPC, EGDT trials, and 17% for the Low-Dose Steroid trial. The moderate and severe skeptic analyses show no clinically meaningful reduction in the risk of death for all trials. If we aim for a RRR >20% or >25%, all probabilities of benefits become lower independent of the degree of skepticism. CONCLUSIONS: Our clinical threshold analysis offers a new bedside tool to be directly applied to the care of patients with severe sepsis. Our results demonstrate that the strength of evidence (statistical and clinical) is weak for all trials, particularly for the Low-Dose Steroid and EGDT trials. It is essential to replicate the results of each of these five clinical trials in confirmatory studies if we want to provide patient care based on scientifically sound evidence. PMID- 18509456 TI - Gene-specific signatures of elevated non-synonymous substitution rates correlate poorly across the Plasmodium genus. AB - BACKGROUND: Comparative genome analyses of parasites allow large scale investigation of selective pressures shaping their evolution. An acute limitation to such analysis of Plasmodium falciparum is that there is only very partial low coverage genome sequence of the most closely related species, the chimpanzee parasite P. reichenowi. However, if orthologous genes have been under similar selective pressures throughout the Plasmodium genus then positive selection on the P. falciparum lineage might be predicted to some extent by analysis of other lineages. PRINCIPAL FINDINGS: Here, three independent pairs of closely related species in different sub-generic clades (P. falciparum and P. reichenowi; P. vivax and P. knowlesi; P. yoelii and P. berghei) were compared for a set of 43 candidate ligand genes considered likely to be under positive directional selection and a set of 102 control genes for which there was no selective hypothesis. The ratios of non-synonymous to synonymous substitutions (dN/dS) were significantly elevated in the candidate ligand genes compared to control genes in each of the three clades. However, the rank order correlation of dN/dS ratios for individual candidate genes was very low, less than the correlation for the control genes. SIGNIFICANCE: The inability to predict positive selection on a gene in one lineage by identifying elevated dN/dS ratios in the orthologue within another lineage needs to be noted, as it reflects that adaptive mutations are generally rare events that lead to fixation in individual lineages. Thus it is essential to complete the genome sequences of particular species of phylogenetic importance, such as P. reichenowi. PMID- 18509457 TI - Ligand specificity of group I biotin protein ligase of Mycobacterium tuberculosis. AB - BACKGROUND: Fatty acids are indispensable constituents of mycolic acids that impart toughness & permeability barrier to the cell envelope of M. tuberculosis. Biotin is an essential co-factor for acetyl-CoA carboxylase (ACC) the enzyme involved in the synthesis of malonyl-CoA, a committed precursor, needed for fatty acid synthesis. Biotin carboxyl carrier protein (BCCP) provides the co-factor for catalytic activity of ACC. METHODOLOGY/PRINCIPAL FINDINGS: BPL/BirA (Biotin Protein Ligase), and its substrate, biotin carboxyl carrier protein (BCCP) of Mycobacterium tuberculosis (Mt) were cloned and expressed in E. coli BL21. In contrast to EcBirA and PhBPL, the approximately 29.5 kDa MtBPL exists as a monomer in native, biotin and bio-5'AMP liganded forms. This was confirmed by molecular weight profiling by gel filtration on Superdex S-200 and Dynamic Light Scattering (DLS). Computational docking of biotin and bio-5'AMP to MtBPL show that adenylation alters the contact residues for biotin. MtBPL forms 11 H-bonds with biotin, relative to 35 with bio-5'AMP. Docking simulations also suggest that bio-5'AMP hydrogen bonds to the conserved 'GRGRRG' sequence but not biotin. The enzyme catalyzed transfer of biotin to BCCP was confirmed by incorporation of radioactive biotin and by Avidin blot. The K(m) for BCCP was approximately 5.2 microM and approximately 420 nM for biotin. MtBPL has low affinity (K(b) = 1.06x10(-6) M) for biotin relative to EcBirA but their K(m) are almost comparable suggesting that while the major function of MtBPL is biotinylation of BCCP, tight binding of biotin/bio-5'AMP by EcBirA is channeled for its repressor activity. CONCLUSIONS/SIGNIFICANCE: These studies thus open up avenues for understanding the unique features of MtBPL and the role it plays in biotin utilization in M. tuberculosis. PMID- 18509458 TI - Cryptic contamination and phylogenetic nonsense. AB - Ancient human DNA has been treated cautiously ever since the problems related to this type of material were exposed in the early 1990s, but as sequential genetic data from ancient specimens have been key components in several evolutionary and ecological studies, interest in ancient human DNA is on the increase again. It is especially tempting to approach archaeological and anthropological questions through this type of material, but DNA from ancient human tissue is notoriously complicated to work with due to the risk of contamination with modern human DNA. Various ways of authenticating results based on ancient human DNA have been developed to circumvent the problems. One commonly used method is to predict what the contamination is expected to look like and then test whether the ancient human DNA fulfils this prediction. If it does, the results are rejected as contamination, while if it does not, they are often considered authentic. We show here that human contamination in ancient material may well deviate from local allele frequencies or the distributions to be found among the laboratory workers and archaeologists. We conclude that it is not reliable to authenticate ancient human DNA solely by showing that it is different from what would be expected from people who have handled the material. PMID- 18509459 TI - Functional implications of plasma membrane condensation for T cell activation. AB - The T lymphocyte plasma membrane condenses at the site of activation but the functional significance of this receptor-mediated membrane reorganization is not yet known. Here we demonstrate that membrane condensation at the T cell activation sites can be inhibited by incorporation of the oxysterol 7 ketocholesterol (7KC), which is known to prevent the formation of raft-like liquid-ordered domains in model membranes. We enriched T cells with 7KC, or cholesterol as control, to assess the importance of membrane condensation for T cell activation. Upon 7KC treatment, T cell antigen receptor (TCR) triggered calcium fluxes and early tyrosine phosphorylation events appear unaltered. However, signaling complexes form less efficiently on the cell surface, fewer phosphorylated signaling proteins are retained in the plasma membrane and actin restructuring at activation sites is impaired in 7KC-enriched cells resulting in compromised downstream activation responses. Our data emphasizes lipids as an important medium for the organization at T cell activation sites and strongly indicates that membrane condensation is an important element of the T cell activation process. PMID- 18509460 TI - Duplicated paralogous genes subject to positive selection in the genome of Trypanosoma brucei. AB - BACKGROUND: Whole genome studies have highlighted duplicated genes as important substrates for adaptive evolution. We have investigated adaptive evolution in this class of genes in the human parasite Trypanosoma brucei, as indicated by the ratio of non-synonymous (amino-acid changing) to synonymous (amino acid retaining) nucleotide substitution rates. METHODOLOGY/PRINCIPAL FINDINGS: We have identified duplicated genes that are most rapidly evolving in this important human parasite. This is the first attempt to investigate adaptive evolution in this species at the codon level. We identify 109 genes within 23 clusters of paralogous gene expansions to be subject to positive selection. CONCLUSIONS/SIGNIFICANCE: Genes identified include surface antigens in both the mammalian and insect host life cycle stage suggesting that competitive interaction is not solely with the adaptive immune system of the mammalian host. Also surface transporters related to drug resistance and genes related to developmental progression are detected. We discuss how adaptive evolution of these genes may highlight lineage specific processes essential for parasite survival. We also discuss the implications of adaptive evolution of these targets for parasite biology and control. PMID- 18509461 TI - Enhancement of both long-term depression induction and optokinetic response adaptation in mice lacking delphilin. AB - In the cerebellum, Delphilin is expressed selectively in Purkinje cells (PCs) and is localized exclusively at parallel fiber (PF) synapses, where it interacts with glutamate receptor (GluR) delta2 that is essential for long-term depression (LTD), motor learning and cerebellar wiring. Delphilin ablation exerted little effect on the synaptic localization of GluRdelta2. There were no detectable abnormalities in cerebellar histology, PC cytology and PC synapse formation in contrast to GluRdelta2 mutant mice. However, LTD induction was facilitated at PF PC synapses in Delphilin mutant mice. Intracellular Ca(2+) required for the induction of LTD appeared to be reduced in the mutant mice, while Ca(2+) influx through voltage-gated Ca(2+) channels and metabotropic GluR1-mediated slow synaptic response were similar between wild-type and mutant mice. We further showed that the gain-increase adaptation of the optokinetic response (OKR) was enhanced in the mutant mice. These findings are compatible with the idea that LTD induction at PF-PC synapses is a crucial rate-limiting step in OKR gain-increase adaptation, a simple form of motor learning. As exemplified in this study, enhancing synaptic plasticity at a specific synaptic site of a neural network is a useful approach to understanding the roles of multiple plasticity mechanisms at various cerebellar synapses in motor control and learning. PMID- 18509462 TI - Brain networks for integrative rhythm formation. AB - BACKGROUND: Performance of externally paced rhythmic movements requires brain and behavioral integration of sensory stimuli with motor commands. The underlying brain mechanisms to elaborate beat-synchronized rhythm and polyrhythms that musicians readily perform may differ. Given known roles in perceiving time and repetitive movements, we hypothesized that basal ganglia and cerebellar structures would have greater activation for polyrhythms than for on-the-beat rhythms. METHODOLOGY/PRINCIPAL FINDINGS: Using functional MRI methods, we investigated brain networks for performing rhythmic movements paced by auditory cues. Musically trained participants performed rhythmic movements at 2 and 3 Hz either at a 1:1 on-the-beat or with a 3:2 or a 2:3 stimulus-movement structure. Due to their prior musical experience, participants performed the 3:2 or 2:3 rhythmic movements automatically. Both the isorhythmic 1:1 and the polyrhythmic 3:2 or 2:3 movements yielded the expected activation in contralateral primary motor cortex and related motor areas and ipsilateral cerebellum. Direct comparison of functional MRI signals obtained during 3:2 or 2:3 and on-the-beat rhythms indicated activation differences bilaterally in the supplementary motor area, ipsilaterally in the supramarginal gyrus and caudate-putamen and contralaterally in the cerebellum. CONCLUSIONS/SIGNIFICANCE: The activated brain areas suggest the existence of an interconnected brain network specific for complex sensory-motor rhythmic integration that might have specificity for elaboration of musical abilities. PMID- 18509463 TI - A population dynamics analysis of the interaction between adaptive regulatory T cells and antigen presenting cells. AB - BACKGROUND: Regulatory T cells are central actors in the maintenance of tolerance of self-antigens or allergens and in the regulation of the intensity of the immune response during infections by pathogens. An understanding of the network of the interaction between regulatory T cells, antigen presenting cells and effector T cells is starting to emerge. Dynamical systems analysis can help to understand the dynamical properties of an interaction network and can shed light on the different tasks that can be accomplished by a network. METHODOLOGY AND PRINCIPAL FINDINGS: We used a mathematical model to describe a interaction network of adaptive regulatory T cells, in which mature precursor T cells may differentiate into either adaptive regulatory T cells or effector T cells, depending on the activation state of the cell by which the antigen was presented. Using an equilibrium analysis of the mathematical model we show that, for some parameters, the network has two stable equilibrium states: one in which effector T cells are strongly regulated by regulatory T cells and another in which effector T cells are not regulated because the regulatory T cell population is vanishingly small. We then simulate different types of perturbations, such as the introduction of an antigen into a virgin system, and look at the state into which the system falls. We find that whether or not the interaction network switches from the regulated (tolerant) state to the unregulated state depends on the strength of the antigenic stimulus and the state from which the network has been perturbed. CONCLUSION/SIGNIFICANCE: Our findings suggest that the interaction network studied in this paper plays an essential part in generating and maintaining tolerance against allergens and self-antigens. PMID- 18509464 TI - Expression of foot-and-mouth disease virus capsid proteins in silkworm baculovirus expression system and its utilization as a subunit vaccine. AB - BACKGROUND: Foot-and-mouth disease (FMD) is a highly contagious disease of livestock that causes severe economic loss in susceptible cloven-hoofed animals. Although the traditional inactivated vaccine has been proved effective, it may lead to a new outbreak of FMD because of either incomplete inactivation of FMDV or the escape of live virus from vaccine production workshop. Thus, it is urgent to develop a novel FMDV vaccine that is safer, more effective and more economical than traditional vaccines. METHODOLOGY AND PRINCIPAL FINDINGS: A recombinant silkworm baculovirus Bm-P12A3C which contained the intact P1-2A and 3C protease coding regions of FMDV Asia 1/HNK/CHA/05 was developed. Indirect immunofluorescence test and sandwich-ELISA were used to verify that Bm-P12A3C could express the target cassette. Expression products from silkworm were diluted to 30 folds and used as antigen to immunize cattle. Specific antibody was induced in all vaccinated animals. After challenge with virulent homologous virus, four of the five animals were completely protected, and clinical symptoms were alleviated and delayed in the remaining one. Furthermore, a PD(50) (50% bovine protective dose) test was performed to assess the bovine potency of the subunit vaccine. The result showed the subunit vaccine could achieve 6.34 PD(50) per dose. CONCLUSION: The results suggest that this strategy might be used to develop the new subunit FMDV vaccine. PMID- 18509465 TI - Age of child, more than HPV type, is associated with clinical course in recurrent respiratory papillomatosis. AB - BACKGROUND: RRP is a devastating disease in which papillomas in the airway cause hoarseness and breathing difficulty. The disease is caused by human papillomavirus (HPV) 6 or 11 and is very variable. Patients undergo multiple surgeries to maintain a patent airway and in order to communicate vocally. Several small studies have been published in which most have noted that HPV 11 is associated with a more aggressive course. METHODOLOGY/PRINCIPAL FINDINGS: Papilloma biopsies were taken from patients undergoing surgical treatment of RRP and were subjected to HPV typing. 118 patients with juvenile-onset RRP with at least 1 year of clinical data and infected with a single HPV type were analyzed. HPV 11 was encountered in 40% of the patients. By our definition, most of the patients in the sample (81%) had run an aggressive course. The odds of a patient with HPV 11 running an aggressive course were 3.9 times higher than that of patients with HPV 6 (Fisher's exact p = 0.017). However, clinical course was more closely associated with age of the patient (at diagnosis and at the time of the current surgery) than with HPV type. Patients with HPV 11 were diagnosed at a younger age (2.4y) than were those with HPV 6 (3.4y) (p = 0.014). Both by multiple linear regression and by multiple logistic regression HPV type was only weakly associated with metrics of disease course when simultaneously accounting for age. CONCLUSIONS/SIGNIFICANCE ABSTRACT: The course of RRP is variable and a quarter of the variability can be accounted for by the age of the patient. HPV 11 is more closely associated with a younger age at diagnosis than it is associated with an aggressive clinical course. These data suggest that there are factors other than HPV type and age of the patient that determine disease course. PMID- 18509466 TI - Case-control cohort study of patients' perceptions of disability in mastocytosis. AB - BACKGROUND: Indolent forms of mastocytosis account for more than 90% of all cases, but the types and type and severity of symptoms and their impact on the quality of life have not been well studied. We therefore performed a case-control cohort study to examine self-reported disability and impact of symptoms on the quality of life in patients with mastocytosis. METHODOLOGY/PRINCIPAL FINDINGS: In 2004, 363 mastocytosis patients and 90 controls in France were asked to rate to their overall disability (OPA score) and the severity of 38 individual symptoms. The latter was used to calculate a composite score (AFIRMM score). Of the 363 respondents, 262 were part of an ongoing pathophysiological study so that the following data were available: World Health Organization classification, standard measures of physical and psychological disability, existence of the D816V KIT mutation, and serum tryptase level. The mean OPA and AFIRMM scores and the standard measures of disability indicated that most mastocytosis patients suffer from disabilities due to the disease. Surprisingly, the patient's measurable and perceived disabilities did not differ according to disease classification or presence or absence of the D816V KIT mutation or an elevated (> or = 20 ng/mL) serum tryptase level. Also, 32 of the 38 AFIRMM symptoms were more common in patients than controls, but there were not substantial differences according to disease classification, presence of the D816V mutation, or the serum tryptase level. CONCLUSIONS: On the basis of these results and for the purposes of treatment, we propose that mastocytosis be first classified as aggressive or indolent and that indolent mastocytosis then be categorized according to the severity of patients' perceived symptoms and their impact on the quality of life. In addition, it appears that mastocytosis patients suffer from more symptoms and greater disability than previously thought, that mastocytosis may therefore be under-diagnosed, and that the symptoms of the indolent forms of mastocytosis might be due more to systemic release of mediators than mast cell burden. PMID- 18509467 TI - The dark side of the salad: Salmonella typhimurium overcomes the innate immune response of Arabidopsis thaliana and shows an endopathogenic lifestyle. AB - Salmonella enterica serovar typhimurium contaminated vegetables and fruits are considerable sources of human infections. Bacteria present in raw plant-derived nutrients cause salmonellosis, the world wide most spread food poisoning. This facultative endopathogen enters and replicates in host cells and actively suppresses host immune responses. Although Salmonella survives on plants, the underlying bacterial infection mechanisms are only poorly understood. In this report we investigated the possibility to use Arabidopsis thaliana as a genetically tractable host system to study Salmonella-plant interactions. Using green fluorescent protein (GFP) marked bacteria, we show here that Salmonella can infect various Arabidopsis tissues and proliferate in intracellular cellular compartments. Salmonella infection of Arabidopsis cells can occur via intact shoot or root tissues resulting in wilting, chlorosis and eventually death of the infected organs. Arabidopsis reacts to Salmonella by inducing the activation of mitogen-activated protein kinase (MAPK) cascades and enhanced expression of pathogenesis related (PR) genes. The induction of defense responses fails in plants that are compromised in ethylene or jasmonic acid signaling or in the MKK3 MPK6 MAPK pathway. These findings demonstrate that Arabidopsis represents a true host system for Salmonella, offering unique possibilities to study the interaction of this human pathogen with plants at the molecular level for developing novel drug targets and addressing current safety issues in human nutrition. PMID- 18509468 TI - Environmental factors contributing to the spread of H5N1 avian influenza in mainland China. AB - BACKGROUND: Since late 2003, highly pathogenic avian influenza (HPAI) outbreaks caused by infection with H5N1 virus has led to the deaths of millions of poultry and more than 10 thousands of wild birds, and as of 18-March 2008, at least 373 laboratory-confirmed human infections with 236 fatalities, have occurred. The unrestrained worldwide spread of this disease has caused great anxiety about the potential of another global pandemic. However, the effect of environmental factors influencing the spread of HPAI H5N1 virus is unclear. METHODOLOGY/PRINCIPAL FINDINGS: A database including incident dates and locations was developed for 128 confirmed HPAI H5N1 outbreaks in poultry and wild birds, as well as 21 human cases in mainland China during 2004-2006. These data, together with information on wild bird migration, poultry densities, and environmental variables (water bodies, wetlands, transportation routes, main cities, precipitation and elevation), were integrated into a Geographical Information System (GIS). A case-control design was used to identify the environmental factors associated with the incidence of the disease. Multivariate logistic regression analysis indicated that minimal distance to the nearest national highway, annual precipitation and the interaction between minimal distance to the nearest lake and wetland, were important predictive environmental variables for the risk of HPAI. A risk map was constructed based on these factors. CONCLUSIONS/SIGNIFICANCE: Our study indicates that environmental factors contribute to the spread of the disease. The risk map can be used to target countermeasures to stop further spread of the HPAI H5N1 at its source. PMID- 18509469 TI - Computational insights on the competing effects of nitric oxide in regulating apoptosis. AB - Despite the establishment of the important role of nitric oxide (NO) on apoptosis, a molecular-level understanding of the origin of its dichotomous pro- and anti-apoptotic effects has been elusive. We propose a new mathematical model for simulating the effects of nitric oxide (NO) on apoptosis. The new model integrates mitochondria-dependent apoptotic pathways with NO-related reactions, to gain insights into the regulatory effect of the reactive NO species N(2)O(3), non-heme iron nitrosyl species (FeL(n)NO), and peroxynitrite (ONOO(-)). The biochemical pathways of apoptosis coupled with NO-related reactions are described by ordinary differential equations using mass-action kinetics. In the absence of NO, the model predicts either cell survival or apoptosis (a bistable behavior) with shifts in the onset time of apoptotic response depending on the strength of extracellular stimuli. Computations demonstrate that the relative concentrations of anti- and pro-apoptotic reactive NO species, and their interplay with glutathione, determine the net anti- or pro-apoptotic effects at long time points. Interestingly, transient effects on apoptosis are also observed in these simulations, the duration of which may reach up to hours, despite the eventual convergence to an anti-apoptotic state. Our computations point to the importance of precise timing of NO production and external stimulation in determining the eventual pro- or anti-apoptotic role of NO. PMID- 18509470 TI - Activation of estrogen receptor-alpha by E2 or EGF induces temporally distinct patterns of large-scale chromatin modification and mRNA transcription. AB - Estrogen receptor-alpha (ER) transcription function is regulated in a ligand dependent (e.g., estradiol, E2) or ligand-independent (e.g., growth factors) manner. Our laboratory seeks to understand these two modes of action. Using a cell line that contains a visible prolactin enhancer/promoter array (PRL-HeLa) regulated by ER, we analyzed ER response to E2 and EGF by quantifying image-based results. Data show differential recruitment of GFP-ER to the array, with the AF1 domain playing a vital role in EGF-mediated responsiveness. Temporal analyses of large-scale chromatin dynamics, and accumulation of array-localized reporter mRNA over 24 hours showed that the EGF response consists of a single pulse of reporter mRNA accumulation concomitant with transient increase in array decondensation. Estradiol induced a novel cyclical pattern of mRNA accumulation with a sustained increase in array decondensation. Collectively, our work shows that there is a stimuli-specific pattern of large-scale chromatin modification and transcript levels by ER. PMID- 18509471 TI - Holidays in the sun and the Caribbean's forgotten burden of neglected tropical diseases. PMID- 18509472 TI - Controlling schistosomiasis: significant decrease of anaemia prevalence one year after a single dose of praziquantel in Nigerian schoolchildren. AB - BACKGROUND: In the framework of the monitoring and evaluation of the Nigerian schistosomiasis and soil-transmitted helminth control programme, a follow-up of children took place in eight sentinel sites. The objective of the study was to assess the evolution of Schistosoma haematobium infection and anaemia in schoolchildren after a single administration of praziquantel (PZQ) and albendazole. METHODS/PRINCIPAL FINDINGS: Pre-treatment examination and follow-up at one year post-treatment of schoolchildren aged 7, 8, and 11 years, including interview, urine examination, ultrasound examination of the urinary tract, and measurement of haemoglobin. Before treatment, the overall prevalence of S. heamatobium infection was 75.4% of the 1,642 enrolled children, and 21.8% of children excreted more than 50 eggs/10 ml urine. Prevalence increased with age. The overall prevalence of anaemia (haemoglobin <11.5 g/dl) was 61.6%, decreasing significantly with increasing age. The mean haemoglobinemia was 11 g/dl. In bivariate analysis, anaemia was significantly more frequent in children infected with S. haematobium, although it was not correlated to the intensity of infection. Anaemia was also associated with micro-haematuria and to kidney distensions. In a sub-sample of 636 children tested for P. falciparum infection, anaemia was significantly more frequent in malaria-infected children. In multivariate analysis, significant predictors of anaemia were P. falciparum infection, kidney distension, and the village. One year after a single-dose praziquantel treatment (administered using the WHO PZQ dose pole) co-administered with albendazole (400 mg single dose) for de-worming, the prevalence of S. haematobium infection was 38%, while the prevalence of anaemia fell to 50.4%. The mean haemoglobinemia showed a statistically significant increase of 0.39 g/dl to reach 11.4 g/dl. Anaemia was no longer associated with S. haematobium or to P. falciparum infections, or to haematuria or ultrasound abnormalities of the urinary tract. CONCLUSIONS: The high prevalence of anaemia in Nigerian children is clearly a result of many factors and not of schistosomiasis alone. Nevertheless, treatment of schistosomiasis and de-worming were followed by a partial, but significant, reduction of anaemia in schoolchildren, not explainable by any other obvious intervention. PMID- 18509473 TI - Irradiation in adulthood as a new model of schizophrenia. AB - BACKGROUND: Epidemiological studies suggest that radiation exposure may be a potential risk factor for schizophrenia in adult humans. Here, we investigated whether adult irradiation in rats caused behavioral abnormalities relevant to schizophrenia. METHODOLOGY/PRINCIPAL FINDINGS: A total dose of 15-Gy irradiation in six fractionations during 3 weeks was exposed to the forebrain including the subventricular zone (SVZ) and subgranular zone (SGZ) with male rats in the prone position. Behavioral, immunohistochemical, and neurochemical studies were performed three months after fractionated ionizing irradiation. Three months after fractionated ionizing irradiation, the total numbers of BrdU-positive cells in both the SVZ and SGZ zones of irradiated rats were significantly lower than those of control (sham-irradiated) rats. Hyperactivity after administration of the dopaminergic agonist methamphetamine, but not the N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine, was significantly enhanced in the irradiated rats although spontaneous locomotion in the irradiated rats was significantly lower than that of controls. Behavioral abnormalities including auditory sensory gating deficits, social interaction deficits, and working memory deficits were observed in the irradiated rats. CONCLUSION/SIGNIFICANCE: The present study suggests that irradiation in adulthood caused behavioral abnormalities relevant to schizophrenia, and that reduction of adult neurogenesis by irradiation may be associated with schizophrenia-like behaviors in rats. PMID- 18509474 TI - High levels of genetic differentiation between Ugandan Glossina fuscipes fuscipes populations separated by Lake Kyoga. AB - BACKGROUND: Glossina fuscipes fuscipes is the major vector of human African trypanosomiasis, commonly referred to as sleeping sickness, in Uganda. In western and eastern Africa, the disease has distinct clinical manifestations and is caused by two different parasites: Trypanosoma brucei rhodesiense and T. b. gambiense. Uganda is exceptional in that it harbors both parasites, which are separated by a narrow 160-km belt. This separation is puzzling considering there are no restrictions on the movement of people and animals across this region. METHODOLOGY AND RESULTS: We investigated whether genetic heterogeneity of G. f. fuscipes vector populations can provide an explanation for this disjunct distribution of the Trypanosoma parasites. Therefore, we examined genetic structuring of G. f. fuscipes populations across Uganda using newly developed microsatellite markers, as well as mtDNA. Our data show that G. f. fuscipes populations are highly structured, with two clearly defined clusters that are separated by Lake Kyoga, located in central Uganda. Interestingly, we did not find a correlation between genetic heterogeneity and the type of Trypanosoma parasite transmitted. CONCLUSIONS: The lack of a correlation between genetic structuring of G. f. fuscipes populations and the distribution of T. b. gambiense and T. b. rhodesiense indicates that it is unlikely that genetic heterogeneity of G. f. fuscipes populations explains the disjunct distribution of the parasites. These results have important epidemiological implications, suggesting that a fusion of the two disease distributions is unlikely to be prevented by an incompatibility between vector populations and parasite. PMID- 18509475 TI - Rabies post-exposure prophylaxis in the Philippines: health status of patients having received purified equine F(ab')(2) fragment rabies immunoglobulin (Favirab). AB - BACKGROUND: Recommended treatment for severe rabies exposure in unvaccinated individuals includes wound cleaning, administration of rabies immunoglobulins (RIG), and rabies vaccination. We conducted a survey of rabies treatment outcomes in the Philippines. METHODS: This was a case series involving 7,660 patients (4 months to 98 years of age) given purified equine RIG (pERIG) at the Research Institute for Tropical Medicine (Muntinlupa, Philippines) from July 2003 to August 2004 following Category II or III exposures. Data on local and systemic adverse reactions (AR) within 28 days and biting animal status were recorded; outcome data were obtained by telephone or home visit 6-29 months post-exposure. RESULTS: Follow-up data were collected for 6,464 patients. Of 151 patients with laboratory-confirmed rabies exposure, 143 were in good health 6-48 months later, seven could not be contacted, and one 4-year-old girl died. Of 16 deaths in total, 14 were unrelated to rabies exposure or treatment. Two deaths were considered PEP failures: the 4-year old girl, who had multiple deep lacerated wounds from a rabid dog of the nape, neck, and shoulders requiring suturing on the day of exposure, and an 8-year-old boy who only received rabies PEP on the day of exposure. CONCLUSIONS: This extensive review of outcomes in persons with Category III exposure shows the recommended treatment schedule at RITM using pERIG is well tolerated, while survival of 143 laboratory-confirmed rabies exposures confirms the intervention efficacy. Two PEP intervention failures demonstrate that sustained education and training is essential in rabies management. PMID- 18509476 TI - A bacterial cytotoxin identifies the RhoA exchange factor Net1 as a key effector in the response to DNA damage. AB - BACKGROUND: Exposure of adherent cells to DNA damaging agents, such as the bacterial cytolethal distending toxin (CDT) or ionizing radiations (IR), activates the small GTPase RhoA, which promotes the formation of actin stress fibers and delays cell death. The signalling intermediates that regulate RhoA activation and promote cell survival are unknown. PRINCIPAL FINDINGS: We demonstrate that the nuclear RhoA-specific Guanine nucleotide Exchange Factor (GEF) Net1 becomes dephosphorylated at a critical inhibitory site in cells exposed to CDT or IR. Expression of a dominant negative Net1 or Net1 knock down by iRNA prevented RhoA activation, inhibited the formation of stress fibers, and enhanced cell death, indicating that Net1 activation is required for this RhoA mediated responses to genotoxic stress. The Net1 and RhoA-dependent signals involved activation of the Mitogen-Activated Protein Kinase p38 and its downstream target MAPK-activated protein kinase 2. SIGNIFICANCE: Our data highlight the importance of Net1 in controlling RhoA and p38 MAPK mediated cell survival in cells exposed to DNA damaging agents and illustrate a molecular pathway whereby chronic exposure to a bacterial toxin may promote genomic instability. PMID- 18509477 TI - iTools: a framework for classification, categorization and integration of computational biology resources. AB - The advancement of the computational biology field hinges on progress in three fundamental directions--the development of new computational algorithms, the availability of informatics resource management infrastructures and the capability of tools to interoperate and synergize. There is an explosion in algorithms and tools for computational biology, which makes it difficult for biologists to find, compare and integrate such resources. We describe a new infrastructure, iTools, for managing the query, traversal and comparison of diverse computational biology resources. Specifically, iTools stores information about three types of resources--data, software tools and web-services. The iTools design, implementation and resource meta-data content reflect the broad research, computational, applied and scientific expertise available at the seven National Centers for Biomedical Computing. iTools provides a system for classification, categorization and integration of different computational biology resources across space-and-time scales, biomedical problems, computational infrastructures and mathematical foundations. A large number of resources are already iTools accessible to the community and this infrastructure is rapidly growing. iTools includes human and machine interfaces to its resource meta-data repository. Investigators or computer programs may utilize these interfaces to search, compare, expand, revise and mine meta-data descriptions of existent computational biology resources. We propose two ways to browse and display the iTools dynamic collection of resources. The first one is based on an ontology of computational biology resources, and the second one is derived from hyperbolic projections of manifolds or complex structures onto planar discs. iTools is an open source project both in terms of the source code development as well as its meta-data content. iTools employs a decentralized, portable, scalable and lightweight framework for long-term resource management. We demonstrate several applications of iTools as a framework for integrated bioinformatics. iTools and the complete details about its specifications, usage and interfaces are available at the iTools web page http://iTools.ccb.ucla.edu. PMID- 18509478 TI - The transmembrane isoform of Plasmodium falciparum MAEBL is essential for the invasion of Anopheles salivary glands. AB - Malaria transmission depends on infective stages in the mosquito salivary glands. Plasmodium sporozoites that mature in midgut oocysts must traverse the hemocoel and invade the mosquito salivary glands in a process thought to be mediated by parasite ligands. MAEBL, a homologue of the transmembrane EBP ligands essential in merozoite invasion, is expressed abundantly in midgut sporozoites. Alternative splicing generates different MAEBL isoforms and so it is unclear what form is functionally essential. To identify the MAEBL isoform required for P. falciparum (NF54) sporozoite invasion of salivary glands, we created knockout and allelic replacements each carrying CDS of a single MAEBL isoform. Only the transmembrane form of MAEBL is essential and is the first P. falciparum ligand validated as essential for invasion of Anopheles salivary glands. MAEBL is the first P. falciparum ligand experimentally determined to be essential for this important step in the life cycle where the vector becomes infectious for transmitting sporozoites to people. With an increasing emphasis on advancing vector-based transgenic methods for suppression of malaria, it is important that this type of study, using modern molecular genetic tools, is done with the agent of the human disease. Understanding what P. falciparum sporozoite ligands are critical for mosquito transmission will help validate targets for vector-based transmission blocking strategies. PMID- 18509479 TI - Unsupervised learning in detection of gene transfer. AB - The tree representation as a model for organismal evolution has been in use since before Darwin. However, with the recent unprecedented access to biomolecular data, it has been discovered that, especially in the microbial world, individual genes making up the genome of an organism give rise to different and sometimes conflicting evolutionary tree topologies. This discovery calls into question the notion of a single evolutionary tree for an organism and gives rise to the notion of an evolutionary consensus tree based on the evolutionary patterns of the majority of genes in a genome embedded in a network of gene histories. Here, we discuss an approach to the analysis of genomic data of multiple genomes using bipartition spectral analysis and unsupervised learning. An interesting observation is that genes within genomes that have evolutionary tree topologies, which are in substantial conflict with the evolutionary consensus tree of an organism, point to possible horizontal gene transfer events which often delineate significant evolutionary events. PMID- 18509480 TI - Reconstruction of a severe open distal humerus fracture with complete loss of medial column by using a free fibular osteocutaneous graft. AB - OBJECTIVE: Open intra-articular fractures of the distal humerus can be associated with considerable bone loss and extensive soft tissue damage. The management of these injuries is quite challenging, and the restoration of elbow anatomy may require multiple bone and soft tissue surgical procedures. The purpose of this case report is to present the option of addressing at the same time a complex skin, muscular, and bone distal humerus defect by using a composite vascularized graft. METHODS: We present a case of a high-energy open fracture of the distal humerus in a polytrauma young patient. Apart from the significant damage of all skin layers and underlying muscle units at the posterolateral side of the elbow, the medial column of the distal humerus (6 cm in length) was completely absent. After patient resuscitation and wound debridement, a free vascularized osteocutaneous fibular graft was used for the reconstruction of the bone defect and the restoration of elbow anatomy. RESULTS: No complications were encountered during the postoperative period, and both bone and soft tissues progressed to sound healing. At 26 months follow-up, the patient had a functional and stable elbow and the Mayo Elbow Performance Score was 85 points, which is equivalent to a good result. CONCLUSIONS: Vascularized osteocutaneous fibular graft could effectively address complex traumatic defects of the elbow joint and enhance the potential for bone healing and early functional recovery. PMID- 18509481 TI - Probabilistic latent variable models as nonnegative factorizations. AB - This paper presents a family of probabilistic latent variable models that can be used for analysis of nonnegative data. We show that there are strong ties between nonnegative matrix factorization and this family, and provide some straightforward extensions which can help in dealing with shift invariances, higher-order decompositions and sparsity constraints. We argue through these extensions that the use of this approach allows for rapid development of complex statistical models for analyzing nonnegative data. PMID- 18509482 TI - Targeting mTOR in HIV-Negative Classic Kaposi's Sarcoma. AB - A 66-year old female with HIV-negative classic Kaposi's sarcoma responded to mTOR targeting by rapamycin. The response was well documented by PET-CT. This case provides supporting evidence that the mTOR pathway may be important in the tumorigenesis of KS and that rapamycin may have activity in this disease. PMID- 18509483 TI - Rapid and sensitive determination of trace chloride ion in drinks using resonance light scattering technique. AB - A resonance light scattering (RLS) technique to determine chloride ions in drinks was developed. Chloride ions were found to bind Ag(+) forming AgCl aggregates that produced intense resonance scattering light. Effects of factors such as acidity, ionic strength, and coexistent interferents on the RLS of AgCl aggregates were investigated. The pH of solution almost did not affect the production of RLS and few foreign species interfered with the detection of chloride ions. The resonance scattering light intensity at the maximum peak of 571 nm was linear to the concentration of chloride ions in the range of 1.42-8.52 ng mL(-1) with a detection limit of 0.71 ng mL(-1). To determine the feasibility of the proposed method, some samples of water and drinks were analyzed. The attained results were in agreement with that of ion-selective electrode method. Good recovery results were also obtained with the range of 94.08-105.63%. The sensitivity and selectivity of the RLS method are high enough to determine trace amounts of chloride ions without any significant interference from high concentration of other components such as common anions and cations. PMID- 18509484 TI - Human granulocytic ehrlichiosis complicating early pregnancy. AB - BACKGROUND: The goal of this case is to review the zoonotic infection, human granulocytic ehrlichiosis, presenting with pyrexia. Case. A 22-year-old multigravid female presented to the emergency department with a painful skin rash, high fever, and severe myalgias. The patient underwent a diagnostic evaluation for zoonotic infections due to her geographical and seasonal risk factors. Treatment of human granulocytic ehrlichiosis was successful though the patient spontaneously aborted presumably due to the severity of the acute illness. CONCLUSION: Treatment of human granulocytic ehrlichiosis in pregnancy presents unique challenges. Management of pyrexia during pregnancy is limited to external cooling in the setting of thrombocytopenia and elevated aminotransferases. Extensive counseling regarding teratogenic potential of medications allows the patient to weigh the pros and cons of treatment. PMID- 18509485 TI - Influence of muscle-tendon wrapping on calculations of joint reaction forces in the equine distal forelimb. AB - The equine distal forelimb is a common location of injuries related to mechanical overload. In this study, a two-dimensional model of the musculoskeletal system of the region was developed and applied to kinematic and kinetic data from walking and trotting horses. The forces in major tendons and joint reaction forces were calculated. The components of the joint reaction forces caused by wrapping of tendons around sesamoid bones were found to be of similar magnitude to the reaction forces between the long bones at each joint. This finding highlighted the importance of taking into account muscle-tendon wrapping when evaluating joint loading in the equine distal forelimb. PMID- 18509486 TI - Citrus genomics. AB - Citrus is one of the most widespread fruit crops globally, with great economic and health value. It is among the most difficult plants to improve through traditional breeding approaches. Currently, there is risk of devastation by diseases threatening to limit production and future availability to the human population. As technologies rapidly advance in genomic science, they are quickly adapted to address the biological challenges of the citrus plant system and the world's industries. The historical developments of linkage mapping, markers and breeding, EST projects, physical mapping, an international citrus genome sequencing project, and critical functional analysis are described. Despite the challenges of working with citrus, there has been substantial progress. Citrus researchers engaged in international collaborations provide optimism about future productivity and contributions to the benefit of citrus industries worldwide and to the human population who can rely on future widespread availability of this health-promoting and aesthetically pleasing fruit crop. PMID- 18509487 TI - PPAR-gamma Thiazolidinedione Agonists and Immunotherapy in the Treatment of Brain Tumors. AB - Thiazolidinediones (TZDs) are selective agonists of the peroxisome proliferator activated receptor (PPAR) gamma, a transcription factor belonging to the superfamily of nuclear hormone receptors. Although activation of PPARgamma by TZDs has been best characterized by its ability to regulate expression of genes associated with lipid metabolism, PPARgamma agonists have other physiological effects including modulating pro- and anti-inflammatory gene expression and inducing apoptosis in several cell types including glioma cells and cell lines. Immunotherapeutic approaches to reducing brain tumors are focused on means to reduce the immunosuppressive responses of tumors which dampen the ability of cytotoxic T-lymphocytes to kill tumors. Initial studies from our lab show that combination of an immunotherapeutic strategy with TZD treatment provides synergistic benefit in animals with implanted tumors. The potential of this combined approach for treatment of brain tumors is reviewed in this report. PMID- 18509488 TI - The creatine transporter gene paralogous at 16p11.2 is expressed in human brain. AB - Autism is a complex neurodevelopmental disorder characterized by impairment of social interaction, language, communication, and stereotyped, repetitive behavior. Genetic predisposition to autism has been demonstrated in families and twin studies. About 5-10% of autism cases are associated with chromosomal abnormalities or monogenic disorders. The identification of genes involved in the origin of autism is expected to increase our understanding of the pathogenesis. We report on the clinical, cytogenetic, and molecular findings in a boy with autism carrying a de novo translocation t(7;16)(p22.1;p11.2). The chromosome 16 breakpoint disrupts the paralogous SLC6A8 gene also called SLC6A10 or CT2. Predicted translation of exons and RT-PCR analysis reveal specific expression of the creatine transporter paralogous in testis and brain. Several studies reported on the role of X-linked creatine transporter mutations in individuals with mental retardation, with or without autism. The existence of disruption in SLC6A8 paralogous gene associated with idiopathic autism suggests that this gene may be involved in the autistic phenotype in our patient. PMID- 18509489 TI - Anticancer Properties of PPARalpha-Effects on Cellular Metabolism and Inflammation. AB - Peroxisome proliferator-activated receptors (PPARs) have lately attracted much attention as therapeutic targets. Previously, PPAR ligands were associated with the treatment of diabetes, hyperlipidemia and cardiovascular diseases, as they modulate the expression of genes regulating glucose and lipid metabolism. Recently, PPAR ligands have been also considered as potential anticancer agents, with relatively low systemic toxicity. The emerging evidence for antiproliferative, proapoptotic, antiinflammatory and potential antimetastatic properties of PPARalpha ligands prompted us to discuss possible roles of PPARalpha in tumor suppression. PPARalpha activation can target cancer cells energy balance by blocking fatty acid synthesis and by promoting fatty acid beta oxidation. In the state of limited nutrient availability, frequently presents in the tumor microenvironment, PPARalpha cooperates with AMP-dependent protein kinase in: (i) repressing oncogenic Akt activity, (ii) inhibiting cell proliferation, and (iii) forcing glycolysis-dependent cancer cells into "metabolic catastrophe." Other potential anticancer effects of PPARalpha include suppression of inflammation, and upregulation of uncoupling proteins (UCPs), which attenuates mitochondrial reactive oxygen species production and cell proliferation. In conclusion, there are strong premises that the low-toxic and well-tolerated PPAR ligands should be considered as new therapeutic agents to fight disseminating cancer, which represents the major challenge for modern medicine and basic research. PMID- 18509490 TI - Worms and the treatment of inflammatory bowel disease: are molecules the answer? AB - The lack of exposure to helminth infections, as a result of improved living standards and medical conditions, may have contributed to the increased incidence of IBD in the developed world. Epidemiological, experimental, and clinical data sustain the idea that helminths could provide protection against IBD. Studies investigating the underlying mechanisms by which helminths might induce such protection have revealed the importance of regulatory pathways, for example, regulatory T-cells. Further investigation on how helminths influence both innate and adaptive immune reactions will shed more light on the complex pathways used by helminths to regulate the hosts immune system. Although therapy with living helminths appears to be effective in several immunological diseases, the disadvantages of a treatment based on living parasites are explicit. Therefore, the identification and characterization of helminth-derived immunomodulatory molecules that contribute to the protective effect could lead to new therapeutic approaches in IBD and other immune diseases. PMID- 18509491 TI - Methylated trivalent arsenic-glutathione complexes are more stable than their arsenite analog. AB - The trivalent arsenic glutathione complexes arsenic triglutathione, methylarsonous diglutathione, and dimethylarsinous glutathione are key intermediates in the mammalian metabolism of arsenite and possibly represent the arsenic species that are transported from the liver to the kidney for urinary excretion. Despite this, the comparative stability of the arsenic-sulfur bonds in these complexes has not been investigated under physiological conditions resembling hepatocyte cytosol. Using size-exclusion chromatography and a glutathione-containing phosphate buffered saline mobile phase (5 or 10 mM glutathione, pH 7.4) in conjunction with an arsenic-specific detector, we chromatographed arsenite, monomethylarsonous acid, and dimethylarsinous acid. The on-column formation of the corresponding arsenic-glutathione complexes between 4 and 37 degrees C revealed that methylated arsenic-glutathione complexes are more stable than arsenic triglutathione. The relevance of these results with regard to the metabolic fate of arsenite in mammals is discussed. PMID- 18509492 TI - Subcellular localization and RNA interference of an RNA methyltransferase gene from silkworm, Bombyx mori. AB - RNA methylation, which is a form of posttranscriptional modification, is catalyzed by S-adenosyl-L-methionone-dependent RNA methyltransterases (RNA MTases). We have identified a novel silkworm gene, BmRNAMTase, containing a 369 bp open reading frame that encodes a putative protein containing 122 amino acid residues and having a molecular weight of 13.88 kd. We expressed a recombinant His-tagged BmRNAMTase in E. coli BL21 (DE3), purified the fusion protein by metal chelation affinity chromatography, and injected a New Zealand rabbit with the purified protein to generate anti-BmRNAMTase polyclonal antibodies. Immunohistochemistry revealed that BmRNAMTase is abundant in the cytoplasm of Bm5 cells. In addition, using RNA interference to reduce the intracellular activity and content of BmRNAMTase, we determined that this cytoplasmic RNA methyltransferase may be involved in preventing cell death in the silkworm. PMID- 18509493 TI - Robust object recognition under partial occlusions using NMF. AB - In recent years, nonnegative matrix factorization (NMF) methods of a reduced image data representation attracted the attention of computer vision community. These methods are considered as a convenient part-based representation of image data for recognition tasks with occluded objects. A novel modification in NMF recognition tasks is proposed which utilizes the matrix sparseness control introduced by Hoyer. We have analyzed the influence of sparseness on recognition rates (RRs) for various dimensions of subspaces generated for two image databases, ORL face database, and USPS handwritten digit database. We have studied the behavior of four types of distances between a projected unknown image object and feature vectors in NMF subspaces generated for training data. One of these metrics also is a novelty we proposed. In the recognition phase, partial occlusions in the test images have been modeled by putting two randomly large, randomly positioned black rectangles into each test image. PMID- 18509494 TI - PPARs in Eye Biology and Disease. PMID- 18509495 TI - History and diagnostic significance of C-peptide. AB - Starting with the epoch-making discovery of proinsulin, C-peptide has played an important interdisciplinary role, both as part of the single-chain precursor molecule and as an individual entity. In the pioneering years, fundamental systematic experiments unravelled new biochemical mechanisms and chemical structures. After the first detection of C-peptide in human serum, it quickly became a most useful independent indicator of insulin biosynthesis and secretion, finding application in a rapidly growing number of clinical investigations. A prerequisite was the development of specific immuno assays for proinsulin and C peptide. Further milestones were: the chemical synthesis of several C-peptides and the accomplishments in the synthesis of proinsulin; the detection of preproinsulin with its bearings on understanding protein biosynthesis; the pioneering role of insulin, proinsulin, C-peptide, and mini-C-peptides in the development of recombinant DNA technology; and the discovery of the enzymes for the endoproteolytic processing of proinsulin into insulin and C-peptide, completing the pathway of biosynthesis. Today, C-peptide continues to serve as a special diagnostic tool in Diabetology and related fields. Thus, its passive role is well established. Evidence for its active role in physiology and pathophysiology is more recent and is subject of the following contributions. PMID- 18509496 TI - Activation and Molecular Targets of Peroxisome Proliferator-Activated Receptor gamma Ligands in Lung Cancer. AB - Lung cancer is the leading cause of cancer death, and five-year survival remains poor, raising the urgency for new treatment strategies. Activation of PPARgamma represents a potential target for both the treatment and prevention of lung cancer. Numerous studies have examined the effect of thiazolidinediones such as rosiglitazone and pioglitazone on lung cancer cells in vitro and in xenograft models. These studies indicate that activation of PPARgamma inhibits cancer cell proliferation as well as invasiveness and metastasis. While activation of PPARgamma can occur by direct binding of pharmacological ligands to the molecule, emerging data indicate that PPARgamma activation can occur through engagement of other signal transduction pathways, including Wnt signaling and prostaglandin production. Data, both from preclinical models and retrospective clinical studies, indicate that activation of PPARgamma may represent an attractive chemopreventive strategy. This article reviews the existing biological and mechanistic experiments focusing on the role of PPARgamma in lung cancer, focusing specifically on nonsmall cell lung cancer. PMID- 18509497 TI - The Role of PPARgamma in Hepatocellular Carcinoma. AB - Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. This cancer develops mainly in cirrhotic patients. The cirrhotic liver is considered to be a preneoplastic organ, suggesting the rationale for cancer prevention. PPARgamma is a nuclear transcription factor whose activation leads to interaction in the metabolism of lipids, insulin sensitization of peripheral cells, anti-inflammatory action. It can also induce differentiation and inhibits proliferation of cancer cells. Until now, data using PPARgamma ligands in HCC have demonstrated mainly in in vitro models that its activation could be due to an antiproliferative effect. PPARgamma ligand administration has also been associated with a diminution of liver fibrosis in animal models, and potentially also on tumoral cell death. Soma data show that the favorable effect of natural and synthetized PPARgamma agonists could also be independent of PPARgamma activation. Furthermore, in some situations, PPARgamma antagonists have also an anticancer effect. Therefore, we can conclude that the link between activation of the PPARgamma pathway and an anticancer activity is suggested but until now not firmly established in HCC. PMID- 18509498 TI - PPARgamma Inhibitors as Novel Tubulin-Targeting Agents. AB - The microtubule-targeting agents (MTAs) are a very successful class of cancer drugs with therapeutic benefits in both hematopoietic and solid tumors. However, resistance to these drugs is a significant problem. Current MTAs bind to microtubules, and/or to their constituent tubulin heterodimers, and affect microtubule polymerization and dynamics. The PPARgamma inhibitor T0070907 can reduce tubulin levels in colorectal cancer cell lines and suppress tumor growth in a murine xenograft model. T0070907 does not alter microtubule polymerization in vitro, and does not appear to work by triggering modulation of tubulin RNA levels subsequent to decreased polymerization. This observation suggests the possible development of antimicrotubule drugs that work by a novel mechanism, and implies the presence of cancer therapeutic targets that have not yet been exploited. This review summarizes what is known about PPARgamma inhibitors and cancer cell death, with emphasis on the tubulin phenotype and PPAR-dependence, and identifies potential mechanisms of action. PMID- 18509499 TI - PPARgamma Agonists: Potential as Therapeutics for Neovascular Retinopathies. AB - The angiogenic, neovascular proliferative retinopathies, proliferative diabetic retinopathy (PDR), and age-dependent macular degeneration (AMD) complicated by choroidal neovascularization (CNV), also termed exudative or "wet" AMD, are common causes of blindness. The antidiabetic thiazolidinediones (TZDs), rosiglitazone, and troglitazone are PPARgamma agonists with demonstrable antiproliferative, and anti-inflammatory effects, in vivo, were shown to ameliorate PDR and CNV in rodent models, implying the potential efficacy of TZDs for treating proliferative retinopathies in humans. Activation of the angiotensin II type 1 receptor (AT1-R) propagates proinflammatory and proliferative pathogenic determinants underlying PDR and CNV. The antihypertensive dual AT1-R blocker (ARB), telmisartan, recently was shown to activate PPARgamma and improve glucose and lipid metabolism and to clinically improve PDR and CNV in rodent models. Therefore, the TZDs and telmisartan, clinically approved antidiabetic and antihypertensive drugs, respectively, may be efficacious for treating and attenuating PDR and CNV humans. Clinical trials are needed to test these possibilities. PMID- 18509501 TI - SNOMED CT survey: an assessment of implementation in EMR/EHR applications. AB - A descriptive study of health information technology (HIT) vendors was conducted to identify which EMR/EHR vendors currently work or anticipate working with SNOMED CT, determine the prevalence of SNOMED CT integration in electronic medical record (EMR) and electronic health record (EHR) products, identify the available and potential future applications for SNOMED CT in EMR/EHR systems, and learn what prompts vendors to include SNOMED CT in EMR/EHR systems. The Web-based survey consisting of 25 questions was fielded in November-December 2006. Seventy two responses were received. The results from this survey on SNOMED CT show a mixed message from respondents with regard to the prevalence of SNOMED CT integration in EMR/EHR products. Those with plans for implementation cited strategic reasons most often. However, HIT vendors who have not yet obtained a SNOMED CT license are waiting for market forces to drive deployment in their systems. Finally, survey respondents currently working with SNOMED CT indicated an expected increase over the next three years in EMR/EHR applications where SNOMED CT will be implemented. PMID- 18509500 TI - C-Peptide effects on renal physiology and diabetes. AB - The C-peptide of proinsulin is important for the biosynthesis of insulin and has for a long time been considered to be biologically inert. Animal studies have shown that some of the renal effects of the C-peptide may in part be explained by its ability to stimulate the Na,K-ATPase activity. Precisely, the C-peptide reduces diabetes-induced glomerular hyperfiltration both in animals and humans, therefore, resulting in regression of fibrosis. The tubular function is also concerned as diabetic animals supplemented with C-peptide exhibit better renal function resulting in reduced urinary sodium waste and protein excretion together with the reduction of the diabetes-induced glomerular hyperfiltration. The tubular effectors of C-peptide were considered to be tubule transporters, but recent studies have shown that biochemical pathways involving cellular kinases and inflammatory pathways may also be important. The matter theory concerning the C-peptide effects is a metabolic one involving the effects of the C-peptide on lipidic metabolic status. This review concentrates on the most convincing data which indicate that the C-peptide is a biologically active hormone for renal physiology. PMID- 18509502 TI - THE PERVERSITY OF INANIMATE OBJECTS: STIMULUS CONTROL BY INCIDENTAL MUSICAL NOTATION. AB - Social cognition research suggests that incidental, environmental stimuli (e.g., business suits) can nonconsciously influence the degree to which behavioral dispositions (e.g., competitiveness) are expressed. Similarly, cognitive research suggests that incidental action-related objects (e.g., hammers) can prime action plans that then affect the speed with which a concurrent, intended action (e.g., power grip) is executed. However, whether incidental stimuli can instigate actions that run counter to one's current goals has yet to be determined. Moving beyond indirect effects, we show that such stimuli can directly cause the expression of undesired actions: Incidental stimuli resembling musical notation caused the systematic expression of unintended key presses in musicians, but not in nonmusicians. Moreover, the effect was found even when targets and distracters bore no apparent perceptual or semantic relation. We discuss the implications of these findings for models of action production and for social-cognitive concepts (e.g., applicability) regarding the limits of nonconscious processing. PMID- 18509503 TI - Effects of Attention on the Strength of Lexical Influences on Speech Perception: Behavioral Experiments and Computational Mechanisms. AB - The effects of lexical context on phonological processing are pervasive and there have been indications that such effects may be modulated by attention. However, attentional modulation in speech processing is neither well-documented nor well understood. Experiment 1 demonstrated attentional modulation of lexical facilitation of speech sound recognition when task and critical stimuli were identical across attention conditions. We propose modulation of lexical activation as a neurophysiologically-plausible computational mechanism that can account for this type of modulation. Contrary to the claims of critics, this mechanism can account for attentional modulation without violating the principle of interactive processing. Simulations of the interactive TRACE model extended to include two different ways of modulating lexical activation showed that each can account for attentional modulation of lexical feedback effects. Experiment 2 tested conflicting predictions from the two implementations and provided evidence that is consistent with bias input as the mechanism of attentional control of lexical activation. PMID- 18509504 TI - Affect is a form of cognition: A neurobiological analysis. AB - In this paper, we suggest that affect meets the traditional definition of "cognition" such that the affect-cognition distinction is phenomenological, rather than ontological. We review how the affect-cognition distinction is not respected in the human brain, and discuss the neural mechanisms by which affect influences sensory processing. As a result of this sensory modulation, affect performs several basic "cognitive" functions. Affect appears to be necessary for normal conscious experience, language fluency, and memory. Finally, we suggest that understanding the differences between affect and cognition will require systematic study of how the phenomenological distinction characterising the two comes about, and why such a distinction is functional. PMID- 18509505 TI - Identification of CD133-positive radioresistant cells in atypical teratoid/rhabdoid tumor. AB - Atypical teratoid/rhabdoid tumor (AT/RT) is an extremely malignant neoplasm in the central nervous system (CNS) which occurs in infancy and childhood. Recent studies suggested that CD133 could be considered a marker for brain cancer stem like cells (CSCs). However, the role of CD133 in AT/RT has never been investigated. Herein we report the isolation of CD133-positive cells (CD133(+)), found to have the potential to differentiate into three germ layer tissues, from tissues of nine AT/RT patients. The migration/invasion/malignancy and radioresistant capabilities of CD133(+) were significantly augmented when compared to CD133(-). The clinical data showed that the amount of CD133(+) in AT/RTs correlated positively with the degree of resistance to radiation therapy. Using cDNA microarray analysis, the genotoxic-response profiles of CD133(+) and CD133(-) irradiated with 10 Gy ionizing radiation (IR) were analyzed 0.5, 2, 6, 12 and 24 h post-IR. We then validated these microarray data and showed increased phosphorylation after IR of p-ATM, p-RAD17, and p-CHX2 as well as increased expression of BCL-2 protein in CD133(+) compared to CD133(-). Furthermore, we found that CD133(+) can effectively resist IR with cisplatin- and/or TRAIL induced apoptosis. Immunohistochemical analysis confirmed the up-regulated expression of p-ATM and BCL-2 proteins in IR-treated CD133(+) xenotransgrafts in SCID mice but not in IR-treated CD133(-). Importantly, the effect of IR in CD133(+) transplanted mice can be significantly improved by a combination of BCL 2 siRNA with debromohymenialdisine, an inhibitor of checkpoint kinases. In sum, this is the first report indicating that CD133(+) AT/RT cells demonstrate the characteristics of CSCs. The IR-resistant and anti-apoptotic properties in CD133(+) may reflect the clinical refractory malignancy of AT/RTs and thus the activated p-ATM pathway and BCL-2 expression in CD133(+) could be possible targets to improve future treatment of deadly diseases like AT/RT. PMID- 18509506 TI - Spatial relational memory requires hippocampal adult neurogenesis. AB - The dentate gyrus of the hippocampus is one of the few regions of the mammalian brain where new neurons are generated throughout adulthood. This adult neurogenesis has been proposed as a novel mechanism that mediates spatial memory. However, data showing a causal relationship between neurogenesis and spatial memory are controversial. Here, we developed an inducible transgenic strategy allowing specific ablation of adult-born hippocampal neurons. This resulted in an impairment of spatial relational memory, which supports a capacity for flexible, inferential memory expression. In contrast, less complex forms of spatial knowledge were unaltered. These findings demonstrate that adult-born neurons are necessary for complex forms of hippocampus-mediated learning. PMID- 18509507 TI - Assessment and treatment of PTSD after a motor vehicle collision: Empirical findings and clinical observations. AB - Individuals who experience a serious motor vehicle accident (MVA) are at increased risk for psychological problems, particularly Posttraumatic Stress Disorder (PTSD). In this article, we review the literature on PTSD among MVA survivors, with particular attention to available instruments to screen for and assess symptomatology of the disorder. Approaches to the treatment of PTSD in this population are reviewed, separated into interventions designed to prevent PTSD in unselected samples, treatment targeting individuals with Acute Stress Disorder that are designed to prevent subsequent development of PTSD, and therapy for individuals with chronic PTSD. Treatment process issues are discussed, in an effort to integrate empirical findings with clinical observations. The empirical literature suggests several approaches to treatment that have good potential outcomes, although continued work is needed to identify factors that predict treatment response, as well as augment individual-based treatment formats. PMID- 18509508 TI - An Accurate Scatter Measurement and Correction Technique for Cone Beam Breast CT Imaging Using Scanning Sampled Measurement (SSM) Technique. AB - We developed and investigated a scanning sampled measurement (SSM) technique for scatter measurement and correction in cone beam breast CT imaging. A cylindrical polypropylene phantom (water equivalent) was mounted on a rotating table in a stationary gantry experimental cone beam breast CT imaging system. A 2-D array of lead beads, with the beads set apart about ~1 cm from each other and slightly tilted vertically, was placed between the object and x-ray source. A series of projection images were acquired as the phantom is rotated 1 degree per projection view and the lead beads array shifted vertically from one projection view to the next. A series of lead bars were also placed at the phantom edge to produce better scatter estimation across the phantom edges. Image signals in the lead beads/bars shadow were used to obtain sampled scatter measurements which were then interpolated to form an estimated scatter distribution across the projection images. The image data behind the lead bead/bar shadows were restored by interpolating image data from two adjacent projection views to form beam-block free projection images. The estimated scatter distribution was then subtracted from the corresponding restored projection image to obtain the scatter removed projection images.Our preliminary experiment has demonstrated that it is feasible to implement SSM technique for scatter estimation and correction for cone beam breast CT imaging. Scatter correction was successfully performed on all projection images using scatter distribution interpolated from SSM and restored projection image data. The resultant scatter corrected projection image data resulted in elevated CT number and largely reduced the cupping effects. PMID- 18509510 TI - RESPONSES OF MALE TROPICAL MOCKINGBIRDS TO VARIATION IN WITHIN-SONG AND BETWEEN SONG VERSATILITY. AB - Despite their large vocal repertoires and otherwise highly versatile singing style, male mockingbirds sometimes sing in a highly repetitive fashion. We conducted a playback experiment to determine the possible signal value of different syllable presentation patterns during simulated male intrusions in the Tropical Mockingbird (Mimus gilvus) testing the hypothesis that more repetitive singing represents a stronger threat and generates a stronger aggressive response. Responses were measured in terms of approach and singing behavior and were analyzed using McGregor's (1992) multivariate method. We also introduce the use of survival analysis for analyzing response variables for which subjects do not perform the behavior in question in at least one of the replicates (known as 'right-censored variables' in the statistical literature). As predicted by theory, experimental subjects responded more aggressively to songs composed of a single note than to variable ones. However, versatility at the between-song level had an opposite effect as high song switching rates generated stronger responses than low ones. Given the lack of a statistical interaction between within-song versatility and switching rate, we conclude that these two parameters may serve independent purposes and possibly transmit different information. We discuss the possibility that the signal value of variation in vocal versatility lies in the mediation of territorial conflicts, the attraction of female partners and/or the mediation of conflicts over access to reproductive females. PMID- 18509509 TI - Vascular endothelial growth factor restores delayed tumor progression in tumors depleted of macrophages. AB - Genetic depletion of macrophages in Polyoma Middle T oncoprotein (PyMT)-induced mammary tumors in mice delayed the angiogenic switch and the progression to malignancy. To determine whether vascular endothelial growth factor A (VEGF-A) produced by tumor-associated macrophages regulated the onset of the angiogenic switch, a genetic approach was used to restore expression of VEGF-A into tumors at the benign stages. This stimulated formation of a high-density vessel network and in macrophage-depleted mice, was followed by accelerated tumor progression. The expression of VEGF-A led to a massive infiltration into the tumor of leukocytes that were mostly macrophages. This study suggests that macrophage produced VEGF regulates malignant progression through stimulating tumor angiogenesis, leukocytic infiltration and tumor cell invasion. PMID- 18509511 TI - Early neurodegeneration progresses independently of microglial activation by heparan sulfate in the brain of mucopolysaccharidosis IIIB mice. AB - BACKGROUND: In mucopolysaccharidosis type IIIB, a lysosomal storage disease causing early onset mental retardation in children, the production of abnormal oligosaccharidic fragments of heparan sulfate is associated with severe neuropathology and chronic brain inflammation. We addressed causative links between the biochemical, pathological and inflammatory disorders in a mouse model of this disease. METHODOLOGY/PRINCIPAL FINDINGS: In cell culture, heparan sulfate oligosaccharides activated microglial cells by signaling through the Toll-like receptor 4 and the adaptor protein MyD88. CD11b positive microglial cells and three-fold increased expression of mRNAs coding for the chemokine MIP1alpha were observed at 10 days in the brain cortex of MPSIIIB mice, but not in MPSIIIB mice deleted for the expression of Toll-like receptor 4 or the adaptor protein MyD88, indicating early priming of microglial cells by heparan sulfate oligosaccharides in the MPSIIIB mouse brain. Whereas the onset of brain inflammation was delayed for several months in doubly mutant versus MPSIIIB mice, the onset of disease markers expression was unchanged, indicating similar progression of the neurodegenerative process in the absence of microglial cell priming by heparan sulfate oligosaccharides. In contrast to younger mice, inflammation in aged MPSIIIB mice was not affected by TLR4/MyD88 deficiency. CONCLUSIONS/SIGNIFICANCE: These results indicate priming of microglia by HS oligosaccharides through the TLR4/MyD88 pathway. Although intrinsic to the disease, this phenomenon is not a major determinant of the neurodegenerative process. Inflammation may still contribute to neurodegeneration in late stages of the disease, albeit independent of TLR4/MyD88. The results support the view that neurodegeneration is primarily cell autonomous in this pediatric disease. PMID- 18509512 TI - Diagnostic and genetic aspects of the Brugada and other inherited arrhythmias syndromes. AB - Doctor Wilde, presenting on behalf of himself and Dr Eckardt, discussed the role of invasive and noninvasive tests for risk stratification of Brugada syndrome. Doctor Hiraoka, presenting on behalf of Y. Yokoyama, M. Takagi, N. Aihara, K. Aonuma, and the Japan Idiopathic Ventricular Fibrillation Study Investigators, further discussed the diagnostic criteria for the Brugada syndrome. Doctor Antzelevitch examined the hypothesis that amplification of spatial dispersion of repolarization in the form of transmural dispersion of repolarization underlies the development of life-threatening ventricular arrhythmias associated with inherited ion channelopathies including the long QT, short QT, and Brugada syndromes. Doctor Corrado discussed the relationship between channelopathies and heart muscle diseases. PMID- 18509513 TI - Synthetic applications of fluorous solid-phase extraction (F-SPE). PMID- 18509514 TI - Waiting Time as a Barrier to Treatment Entry: Perceptions of Substance Users. AB - Many substance users report that they experience multiple barriers that produce significant challenges to linking with treatment services. Being on a waiting list is frequently mentioned as a barrier, leading some people to give up on treatment and to continue using, while prompting others to view sobriety during the waiting period as proof they do not need treatment. This ethnographic study examines the views that 52 substance users have of the waiting time before treatment and the strategies they created to overcome it. Understanding how substance users react to waiting time itself and in relation to other barriers can lead to services that are effective in encouraging treatment linkage. PMID- 18509516 TI - Space-time visualization and analysis in the Cancer Atlas Viewer. AB - This article describes the Cancer Atlas Viewer: free, downloadable software for the exploration of United States cancer mortality data. We demonstrate the software by exploring spatio-temporal patterns in colon cancer mortality rates for African-American and white females and males in the southeastern United States over the period 1970-1995. We compare the results of two cluster statistics: the local Moran and the local G*, through time.. Overall, the two statistics reach similar conclusions for most locations, although where they disagree reveals some interesting patterns in the data. There are only two persistent clusters of colon cancer mortality, and these are clusters of low values. PMID- 18509515 TI - A Social Neuroscience Perspective on Adolescent Risk-Taking. AB - This article proposes a framework for theory and research on risk-taking that is informed by developmental neuroscience. Two fundamental questions motivate this review. First, why does risk-taking increase between childhood and adolescence? Second, why does risk-taking decline between adolescence and adulthood? Risk taking increases between childhood and adolescence as a result of changes around the time of puberty in the brain's socio-emotional system leading to increased reward-seeking, especially in the presence of peers, fueled mainly by a dramatic remodeling of the brain's dopaminergic system. Risk-taking declines between adolescence and adulthood because of changes in the brain's cognitive control system - changes which improve individuals' capacity for self-regulation. These changes occur across adolescence and young adulthood and are seen in structural and functional changes within the prefrontal cortex and its connections to other brain regions. The differing timetables of these changes make mid-adolescence a time of heightened vulnerability to risky and reckless behavior. PMID- 18509518 TI - Adolescent perceptions of friendship and their associations with individual adjustment. AB - This study of 282 dyads examines early- and mid-adolescents' perceptions of friendship quality and their association with daily disagreements, self- and mother reports of behaviour problems, and school grades. Actor and partner analyses identify unique associations between perceptions of friendship quality and perceptions of daily conflict. Actor effects reveal links between friendship negativity and self-perceptions of conflict affective intensity, relationship impact, post-conflict interaction, and post-conflict separation, and between friendship positivity and self-perceptions of relationship impact. Partner effects reveal links between friendship negativity and partner perceptions of conflict outcomes. Perceptions of relationship quality were also associated with self- and mother reports of behaviour problems and with school grades, such that individual and dyadic views of friendship negativity were linked to detrimental outcomes. The worst outcomes tended to be reserved for dyads in which one or both friends reported high levels of relationship negativity. PMID- 18509519 TI - Analysis of Cardiac Function from MR Images. PMID- 18509520 TI - Is bird song a reliable signal of aggressive intent? AB - In this commentary, we discuss recent experiments on the reliability of bird song as a signal of aggressive intent during territorial conflicts. We outline relevant theoretical views on honest signaling, highlighting the vulnerability handicap hypothesis as a possible explanation for soft song's reliability in predicting attack. We also sketch possible methods of testing whether soft song agrees with key predictions of the vulnerability handicap hypothesis. Finally, we suggest possible empirical refinements that may be useful in future studies of signals of intent, both in birds and in animals broadly. In particular, we argue that future studies of intent should strive to incorporate the following elements into their experimental design: (1) multi-modal signal components, (2) interaction dynamics, and (3) minimal time intervals. Simulated exchanges using dynamically-interactive models may provide a powerful means of incorporating all three of these design features simultaneously. PMID- 18509521 TI - Estimation of relevant variables on high-dimensional biological patterns using iterated weighted kernel functions. AB - BACKGROUND: The analysis of complex proteomic and genomic profiles involves the identification of significant markers within a set of hundreds or even thousands of variables that represent a high-dimensional problem space. The occurrence of noise, redundancy or combinatorial interactions in the profile makes the selection of relevant variables harder. METHODOLOGY/PRINCIPAL FINDINGS: Here we propose a method to select variables based on estimated relevance to hidden patterns. Our method combines a weighted-kernel discriminant with an iterative stochastic probability estimation algorithm to discover the relevance distribution over the set of variables. We verified the ability of our method to select predefined relevant variables in synthetic proteome-like data and then assessed its performance on biological high-dimensional problems. Experiments were run on serum proteomic datasets of infectious diseases. The resulting variable subsets achieved classification accuracies of 99% on Human African Trypanosomiasis, 91% on Tuberculosis, and 91% on Malaria serum proteomic profiles with fewer than 20% of variables selected. Our method scaled-up to dimensionalities of much higher orders of magnitude as shown with gene expression microarray datasets in which we obtained classification accuracies close to 90% with fewer than 1% of the total number of variables. CONCLUSIONS: Our method consistently found relevant variables attaining high classification accuracies across synthetic and biological datasets. Notably, it yielded very compact subsets compared to the original number of variables, which should simplify downstream biological experimentation. PMID- 18509522 TI - FAST ANALYTICAL RECONSTRUCTION OF GATED CARDIAC SPECT WITH NON-UNIFORM ATTENUATION COMPENSATION. AB - Conventionally, the inverse problem of gated cardiac SPECT is solved by reconstructing the images frame-by-frame, ignoring the inter-frame correlation along the time dimension. To compensate for the non-uniform attenuation for quantitative cardiac imaging, iterative image reconstruction has been a choice which could utilize an a priori constraint on the inter-frame correlation for a penalized maximum likelihood (ML) solution. However, iterative image reconstruction in the 4D space involves intensive computations. In this paper, an efficient method for 4D gated SPECT reconstruction is developed based on Karhune Loeve (KL) transform and Novikov's inverse formula. The temporal KL transform is first applied on the data sequence to de-correlate the inter-frame correlation and then the 3D principal components in the KL domain are reconstructed frame-by frame using Novikov's inverse formula with non-uniform attenuation compensation. Finally an inverse KL transform is performed to obtain quantitatively reconstructed 4D images in the original space. With the proposed method, 4D reconstruction can be achieved at a reasonable computational cost. The results from computer simulations are very encouraging as compared to conventional frame by-frame filtered back-projection and iterative ordered-subsets ML reconstructions. By discarding high-order KL components for further noise reduction, the computation time could be further reduced. PMID- 18509523 TI - The binary protein interactome of Treponema pallidum--the syphilis spirochete. AB - Protein interaction networks shed light on the global organization of proteomes but can also place individual proteins into a functional context. If we know the function of bacterial proteins we will be able to understand how these species have adapted to diverse environments including many extreme habitats. Here we present the protein interaction network for the syphilis spirochete Treponema pallidum which encodes 1,039 proteins, 726 (or 70%) of which interact via 3,649 interactions as revealed by systematic yeast two-hybrid screens. A high confidence subset of 991 interactions links 576 proteins. To derive further biological insights from our data, we constructed an integrated network of proteins involved in DNA metabolism. Combining our data with additional evidences, we provide improved annotations for at least 18 proteins (including TP0004, TP0050, and TP0183 which are suggested to be involved in DNA metabolism). We estimate that this "minimal" bacterium contains on the order of 3,000 protein interactions. Profiles of functional interconnections indicate that bacterial proteins interact more promiscuously than eukaryotic proteins, reflecting the non compartmentalized structure of the bacterial cell. Using our high-confidence interactions, we also predict 417,329 homologous interactions ("interologs") for 372 completely sequenced genomes and provide evidence that at least one third of them can be experimentally confirmed. PMID- 18509525 TI - Concerted action is needed to tackle liver fluke infections in Asia. PMID- 18509524 TI - Life-cycle and genome of OtV5, a large DNA virus of the pelagic marine unicellular green alga Ostreococcus tauri. AB - Large DNA viruses are ubiquitous, infecting diverse organisms ranging from algae to man, and have probably evolved from an ancient common ancestor. In aquatic environments, such algal viruses control blooms and shape the evolution of biodiversity in phytoplankton, but little is known about their biological functions. We show that Ostreococcus tauri, the smallest known marine photosynthetic eukaryote, whose genome is completely characterized, is a host for large DNA viruses, and present an analysis of the life-cycle and 186,234 bp long linear genome of OtV5. OtV5 is a lytic phycodnavirus which unexpectedly does not degrade its host chromosomes before the host cell bursts. Analysis of its complete genome sequence confirmed that it lacks expected site-specific endonucleases, and revealed the presence of 16 genes whose predicted functions are novel to this group of viruses. OtV5 carries at least one predicted gene whose protein closely resembles its host counterpart and several other host-like sequences, suggesting that horizontal gene transfers between host and viral genomes may occur frequently on an evolutionary scale. Fifty seven percent of the 268 predicted proteins present no similarities with any known protein in Genbank, underlining the wealth of undiscovered biological diversity present in oceanic viruses, which are estimated to harbour 200Mt of carbon. PMID- 18509526 TI - Human T-lymphotropic virus-1 visualized at the virological synapse by electron tomography. AB - Human T-lymphotropic virus 1 (HTLV-1) is transmitted directly between cells via an organized cell-cell contact called a virological synapse (VS). The VS has been studied by light microscopy, but the ultrastructure of the VS and the nature of the transmitted viral particle have remained unknown. Cell-free enveloped virions of HTLV-1 are undetectable in the serum of individuals infected with the human T lymphotropic virus 1 (HTLV-1) and during in vitro culture of naturally infected lymphocytes. However, the viral envelope protein is required for infectivity of HTLV-1, suggesting that complete, enveloped HTLV-1 virions are transferred across the synapse. Here, we use electron tomography combined with immunostaining of viral protein to demonstrate the presence of enveloped HTLV-1 particles within the VS formed between naturally infected lymphocytes. We show in 3D that HTLV-1 particles can be detected in multiple synaptic clefts at different locations simultaneously within the same VS. The synaptic clefts are surrounded by the tightly apposed plasma membranes of the two cells. HTLV-1 virions can contact the recipient cell membrane before detaching from the infected cell. The results show that the HTLV-1 virological synapse that forms spontaneously between lymphocytes of HTLV-1 infected individuals allows direct cell-cell transmission of the virus by triggered, directional release of enveloped HTLV-1 particles into confined intercellular spaces. PMID- 18509527 TI - Elevated expression of phospholipid transfer protein in bone marrow derived cells causes atherosclerosis. AB - BACKGROUND: Phospholipid transfer protein (PLTP) is expressed by various cell types. In plasma, it is associated with high density lipoproteins (HDL). Elevated levels of PLTP in transgenic mice result in decreased HDL and increased atherosclerosis. PLTP is present in human atherosclerotic lesions, where it seems to be macrophage derived. The aim of the present study is to evaluate the atherogenic potential of macrophage derived PLTP. METHODS AND FINDINGS: Here we show that macrophages from human PLTP transgenic mice secrete active PLTP. Subsequently, we performed bone marrow transplantations using either wild type mice (PLTPwt/wt), hemizygous PLTP transgenic mice (huPLTPtg/wt) or homozygous PLTP transgenic mice (huPLTPtg/tg) as donors and low density lipoprotein receptor deficient mice (LDLR-/-) as acceptors, in order to establish the role of PLTP expressed by bone marrow derived cells in diet-induced atherogenesis. Atherosclerosis was increased in the huPLTPtg/wt-->LDLR-/- mice (2.3-fold) and even further in the huPLTPtg/tg-->LDLR-/- mice (4.5-fold) compared with the control PLTPwt/wt-->LDLR-/- mice (both P<0.001). Plasma PLTP activity levels and non-HDL cholesterol were increased and HDL cholesterol decreased compared with controls (all P<0.01). PLTP was present in atherosclerotic plaques in the mice as demonstrated by immunohistochemistry and appears to co-localize with macrophages. Isolated macrophages from PLTP transgenic mice do not show differences in cholesterol efflux or in cytokine production. Lipopolysaccharide activation of macrophages results in increased production of PLTP. This effect was strongly amplified in PLTP transgenic macrophages. CONCLUSIONS: We conclude that PLTP expression by bone marrow derived cells results in atherogenic effects on plasma lipids, increased PLTP activity, high local PLTP protein levels in the atherosclerotic lesions and increased atherosclerotic lesion size. PMID- 18509528 TI - Prolonged visual experience in adulthood modulates holistic face perception. AB - BACKGROUND: Using the well-known composite illusion as a marker of the holistic perception of faces, we tested how prolonged visual experience with a specific population of faces (4- to 6-year-old children) modulates the face perception system in adulthood. METHODOLOGY/PRINCIPAL FINDINGS: We report a face composite effect that is larger for adult than children faces in a group of adults without experience with children faces ("children-face novices"), while it is of equal magnitude for adults and children faces in a population of preschool teachers ("children-face experts"). When considering preschool teachers only, we observed a significant correlation between the number of years of experience with children faces and the differential face composite effect between children and adults faces. Participants with at least 10 years of qualitative experience with children faces had a larger composite face effect for children than adult faces. CONCLUSIONS/SIGNIFICANCE: Overall, these observations indicate that even in adulthood face processes can be reshaped qualitatively, presumably to facilitate efficient processing of the differential morphological features of the frequently encountered population of faces. PMID- 18509529 TI - Resistance of Foxp3+ regulatory T cells to Nur77-induced apoptosis promotes allograft survival. AB - The NR4A nuclear receptor family member Nur77 (NR4A1) promotes thymocyte apoptosis during negative selection of autoreactive thymocytes, but may also function in mature extrathymic T cells. We studied the effects of over-expression of Nur77 on the apoptosis of murine peripheral T cells, including thymic-derived Foxp3+ regulatory (Treg) cells. Overexpression of Nur77 in the T cell lineage decreased numbers of peripheral CD4 and CD8 T cells by approximately 80% compared to wild-type (WT) mice. However, the proportions of Treg cells were markedly increased in the thymus (61% of CD4+Foxp3+ singly positive thymocytes vs. 8% in WT) and secondary lymphoid organs (40-50% of CD4+Foxp3+ T cells vs. 7-8% in WT) of Nur77 transgenic (Nur77Tg) mice, and immunoprecipitation studies showed Nur77 was associated with a recently identified HDAC7/Foxp3 transcriptional complex. Upon activation through the T cell receptor in vitro or in vivo, Nur77Tg T cells showed only marginally decreased proliferation but significantly increased apoptosis. Fully allogeneic cardiac grafts transplanted to Nur77Tg mice survived long-term with well-preserved structure, and recipient splenocytes showed markedly enhanced apoptosis and greatly reduced anti-donor recall responses. Allografts in Nur77Tg recipients had significantly increased expression of multiple Treg-associated genes, including Foxp3, Foxp1, Tip60 and HDAC9. Allograft rejection was restored by CD25 monoclonal antibody therapy, indicating that allograft acceptance was dependent upon Treg function in Nur77Tg recipients. These data show that compared to conventional CD4 and CD8 T cells, Foxp3+ Tregs are relatively resistant to Nur77-mediated apoptosis, and that tipping the balance between the numbers of Tregs and responder T cells in the early period post-transplantation can determine the fate of the allograft. Hence, induced expression of Nur77 might be a novel means to achieve long-term allograft survival. PMID- 18509530 TI - M867, a novel selective inhibitor of caspase-3 enhances cell death and extends tumor growth delay in irradiated lung cancer models. AB - BACKGROUND: Lung cancer remains the leading cause of cancer death worldwide. Radioresistance of lung cancer cells results in unacceptable rate of loco regional failure. Although radiation is known to induce apoptosis, our recent study showed that knockdown of pro-apoptotic proteins Bak and Bax resulted in an increase in autophagic cell death and lung cancer radiosensitivity in vitro. To further explore the potential of apoptosis inhibition as a way to sensitize lung cancer for therapy, we tested M867, a novel chemical and reversible caspase-3 inhibitor, in combination with ionizing radiation in vivo and in vitro. METHODS AND FINDINGS: M867 reduced clonogenic survival in H460 lung cancer cells (DER = 1.27, p = 0.007) compared to the vehicle-treated treated cells. We found that administration of M867 with ionizing radiation in an in vivo mouse hind limb lung cancer model was well tolerated, and produced a significant tumor growth delay compared to radiation alone. A dramatic decrease in tumor vasculature was observed with M867 and radiation using von Willebrand factor staining. In addition, Ki67 index showed >5-fold reduction of tumor proliferation in the combination therapy group, despite the reduced levels of apoptosis observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Radiosensitizing effect of M867 through inhibiting caspases was validated using caspase-3/-7 double-knockout (DKO) mouse embryonic fibroblasts (MEF) cell model. Consistent with our previous study, autophagy contributed to the mechanism of increased cell death, following inhibition of apoptosis. In addition, matrigel assay showed a decrease in in vitro endothelial tubule formation during the M867/radiation combination treatment. CONCLUSIONS: M867 enhances the cytotoxic effects of radiation on lung cancer and its vasculature both in vitro and in vivo. M867 has the potential to prolong tumor growth delay by inhibiting tumor proliferation. Clinical trials are needed to determine the potential of this combination therapy in patients with locally advanced lung cancer. PMID- 18509531 TI - Metalloprotease meprin beta in rat kidney: glomerular localization and differential expression in glomerulonephritis. AB - Meprin (EC 3.4.24.18) is an oligomeric metalloendopeptidase found in microvillar membranes of kidney proximal tubular epithelial cells. Here, we present the first report on the expression of meprin beta in rat glomerular epithelial cells and suggest a potential involvement in experimental glomerular disease. We detected meprin beta in glomeruli of immunostained rat kidney sections on the protein level and by quantitative RT-PCR of laser-capture microdissected glomeruli on the mRNA level. Using immuno-gold staining we identified the membrane of podocyte foot processes as the main site of meprin beta expression. The glomerular meprin beta expression pattern was altered in anti-Thy 1.1 and passive Heymann nephritis (PHN). In addition, the meprin beta staining pattern in the latter was reminiscent of immunostaining with the sheep anti-Fx1A antiserum, commonly used in PHN induction. Using Western blot and immunoprecipitation assays we demonstrated that meprin beta is recognized by Fx1A antiserum and may therefore represent an auto-antigen in PHN. In anti-Thy 1.1 glomerulonephritis we observed a striking redistribution of meprin beta in tubular epithelial cells from the apical to the basolateral side and the cytosol. This might point to an involvement of meprin beta in this form of glomerulonephritis. PMID- 18509532 TI - Plasmodium falciparum antigens on the surface of the gametocyte-infected erythrocyte. AB - BACKGROUND: The asexual blood stages of the human malaria parasite Plasmodium falciparum produce highly immunogenic polymorphic antigens that are expressed on the surface of the host cell. In contrast, few studies have examined the surface of the gametocyte-infected erythrocyte. METHODOLOGY/PRINCIPAL FINDINGS: We used flow cytometry to detect antibodies recognising the surface of live cultured erythrocytes infected with gametocytes of P. falciparum strain 3D7 in the plasma of 200 Gambian children. The majority of children had been identified as carrying gametocytes after treatment for malaria, and each donated blood for mosquito feeding experiments. None of the plasma recognised the surface of erythrocytes infected with developmental stages of gametocytes (I-IV), but 66 of 194 (34.0%) plasma contained IgG that recognised the surface of erythrocytes infected with mature (stage V) gametocytes. Thirty-four (17.0%) of 200 plasma tested recognised erythrocytes infected with trophozoites and schizonts, but there was no association with recognition of the surface of gametocyte-infected erythrocytes (odds ratio 1.08, 95% C.I. 0.434-2.57; P = 0.851). Plasma antibodies with the ability to recognise gametocyte surface antigens (GSA) were associated with the presence of antibodies that recognise the gamete antigen Pfs 230, but not Pfs48/45. Antibodies recognising GSA were associated with donors having lower gametocyte densities 4 weeks after antimalarial treatment. CONCLUSIONS/SIGNIFICANCE: We provide evidence that GSA are distinct from antigens detected on the surface of asexual 3D7 parasites. Our findings suggest a novel strategy for the development of transmission-blocking vaccines. PMID- 18509533 TI - Iron(III)-salophene: an organometallic compound with selective cytotoxic and anti proliferative properties in platinum-resistant ovarian cancer cells. AB - BACKGROUND: In this pioneer study to the biological activity of organometallic compound Iron(III)-salophene (Fe-SP) the specific effects of Fe-SP on viability, morphology, proliferation, and cell-cycle progression on platinum-resistant ovarian cancer cell lines were investigated. METHODOLOGY/PRINCIPAL FINDINGS: Fe SP displayed selective cytotoxicity against SKOV-3 and OVCAR-3 (ovarian epithelial adenocarcinoma) cell lines at concentrations between 100 nM and 1 microM, while the viability of HeLa cells (epithelial cervix adenocarcinoma) or primary lung or skin fibroblasts was not affected. SKOV-3 cells in contrast to fibroblasts after treatment with Fe-SP revealed apparent hallmarks of apoptosis including densely stained nuclear granular bodies within fragmented nuclei, highly condensed chromatin and chromatin fragmentation. Fe-SP treatment led to the activation of markers of the extrinsic (Caspase-8) and intrinsic (Caspase-9) pathway of apoptosis as well as of executioner Caspase-3 while PARP-1 was deactivated. Fe-SP exerted effects as an anti-proliferative agent with an IC(50) value of 300 nM and caused delayed progression of cells through S-phase phase of the cell cycle resulting in a complete S-phase arrest. When intra-peritoneally applied to rats Fe-SP did not show any systemic toxicity at concentrations that in preliminary trials were determined to be chemotherapeutic relevant doses in a rat ovarian cancer cell model. CONCLUSION/SIGNIFICANCE: The present report suggests that Fe-SP is a potent growth-suppressing agent in vitro for cell lines derived from ovarian cancer and a potential therapeutic drug to treat such tumors in vivo. PMID- 18509534 TI - Toward the assessment of food toxicity for celiac patients: characterization of monoclonal antibodies to a main immunogenic gluten peptide. AB - BACKGROUND AND AIMS: Celiac disease is a permanent intolerance to gluten prolamins from wheat, barley, rye and, in some patients, oats. Partially digested gluten peptides produced in the digestive tract cause inflammation of the small intestine. High throughput, immune-based assays using monoclonal antibodies specific for these immunotoxic peptides would facilitate their detection in food and enable monitoring of their enzymatic detoxification. Two monoclonal antibodies, G12 and A1, were developed against a highly immunotoxic 33-mer peptide. The potential of each antibody for quantifying food toxicity for celiac patients was studied. METHODS: Epitope preferences of G12 and A1 antibodies were determined by ELISA with gluten-derived peptide variants of recombinant, synthetic or enzymatic origin. RESULTS: The recognition sequences of G12 and A1 antibodies were hexameric and heptameric epitopes, respectively. Although G12 affinity for the 33-mer was superior to A1, the sensitivity for gluten detection was higher for A1. This observation correlated to the higher number of A1 epitopes found in prolamins than G12 epitopes. Activation of T cell from gluten digested by glutenases decreased equivalently to the detection of intact peptides by A1 antibody. Peptide recognition of A1 included gliadin peptides involved in the both the adaptive and innate immunological response in celiac disease. CONCLUSIONS: The sensitivity and epitope preferences of the A1 antibody resulted to be useful to detect gluten relevant peptides to infer the potential toxicity of food for celiac patients as well as to monitor peptide modifications by transglutaminase 2 or glutenases. PMID- 18509535 TI - Function of the Shaw potassium channel within the Drosophila circadian clock. AB - BACKGROUND: In addition to the molecular feedback loops, electrical activity has been shown to be important for the function of networks of clock neurons in generating rhythmic behavior. Most studies have used over-expression of foreign channels or pharmacological manipulations that alter membrane excitability. In order to determine the cellular mechanisms that regulate resting membrane potential (RMP) in the native clock of Drosophila we modulated the function of Shaw, a widely expressed neuronal potassium (K(+)) channel known to regulate RMP in Drosophila central neurons. METHODOLOGY/PRINCIPAL FINDINGS: We show that Shaw is endogenously expressed in clock neurons. Differential use of clock gene promoters was employed to express a range of transgenes that either increase or decrease Shaw function in different clusters of clock neurons. Under LD conditions, increasing Shaw levels in all clock neurons (LNv, LNd, DN(1), DN(2) and DN(3)), or in subsets of clock neurons (LNd and DNs or DNs alone) increases locomotor activity at night. In free-running conditions these manipulations result in arrhythmic locomotor activity without disruption of the molecular clock. Reducing Shaw in the DN alone caused a dramatic lengthening of the behavioral period. Changing Shaw levels in all clock neurons also disrupts the rhythmic accumulation and levels of Pigment Dispersing Factor (PDF) in the dorsal projections of LNv neurons. However, changing Shaw levels solely in LNv neurons had little effect on locomotor activity or rhythmic accumulation of PDF. CONCLUSIONS/SIGNIFICANCE: Based on our results it is likely that Shaw modulates pacemaker and output neuronal electrical activity that controls circadian locomotor behavior by affecting rhythmic release of PDF. The results support an important role of the DN clock neurons in Shaw-mediated control of circadian behavior. In conclusion, we have demonstrated a central role of Shaw for coordinated and rhythmic output from clock neurons. PMID- 18509536 TI - A role for cytoplasmic PML in cellular resistance to viral infection. AB - PML gene was discovered as a fusion partner with retinoic acid receptor (RAR) alpha in the t(15:17) chromosomal translocation associated with acute promyelocytic leukemia (APL). Nuclear PML protein has been implicated in cell growth, tumor suppression, apoptosis, transcriptional regulation, chromatin remodeling, DNA repair, and anti-viral defense. The localization pattern of promyelocytic leukemia (PML) protein is drastically altered during viral infection. This alteration is traditionally viewed as a viral strategy to promote viral replication. Although multiple PML splice variants exist, we demonstrate that the ratio of a subset of cytoplasmic PML isoforms lacking exons 5 & 6 is enriched in cells exposed to herpes simplex virus-1 (HSV-1). In particular, we demonstrate that a PML isoform lacking exons 5 & 6, called PML Ib, mediates the intrinsic cellular defense against HSV-1 via the cytoplasmic sequestration of the infected cell protein (ICP) 0 of HSV-1. The results herein highlight the importance of cytoplasmic PML and call for an alternative, although not necessarily exclusive, interpretation regarding the redistribution of PML that is seen in virally infected cells. PMID- 18509537 TI - Evidence of authentic DNA from Danish Viking Age skeletons untouched by humans for 1,000 years. AB - BACKGROUND: Given the relative abundance of modern human DNA and the inherent impossibility for incontestable proof of authenticity, results obtained on ancient human DNA have often been questioned. The widely accepted rules regarding ancient DNA work mainly affect laboratory procedures, however, pre-laboratory contamination occurring during excavation and archaeological-/anthropological handling of human remains as well as rapid degradation of authentic DNA after excavation are major obstacles. METHODOLOGY/PRINCIPAL FINDINGS: We avoided some of these obstacles by analyzing DNA from ten Viking Age subjects that at the time of sampling were untouched by humans for 1,000 years. We removed teeth from the subjects prior to handling by archaeologists and anthropologists using protective equipment. An additional tooth was removed after standard archaeological and anthropological handling. All pre-PCR work was carried out in a "clean- laboratory" dedicated solely to ancient DNA work. Mitochondrial DNA was extracted and overlapping fragments spanning the HVR-1 region as well as diagnostic sites in the coding region were PCR amplified, cloned and sequenced. Consistent results were obtained with the "unhandled" teeth and there was no indication of contamination, while the latter was the case with half of the "handled" teeth. The results allowed the unequivocal assignment of a specific haplotype to each of the subjects, all haplotypes being compatible in their character states with a phylogenetic tree drawn from present day European populations. Several of the haplotypes are either infrequent or have not been observed in modern Scandinavians. The observation of haplogroup I in the present study (<2% in modern Scandinavians) supports our previous findings of a pronounced frequency of this haplogroup in Viking and Iron Age Danes. CONCLUSION: The present work provides further evidence that retrieval of ancient human DNA is a possible task provided adequate precautions are taken and well-considered sampling is applied. PMID- 18509538 TI - General practice and pandemic influenza: a framework for planning and comparison of plans in five countries. AB - BACKGROUND: Although primary health care, and in particular, general practice will be at the frontline in the response to pandemic influenza, there are no frameworks to guide systematic planning for this task or to appraise available plans for their relevance to general practice. We aimed to develop a framework that will facilitate planning for general practice, and used it to appraise pandemic plans from Australia, England, USA, New Zealand and Canada. METHODOLOGY/PRINCIPAL FINDINGS: We adapted the Haddon matrix to develop the framework, populating its cells through a multi-method study that incorporated the peer-reviewed and grey literature, interviews with general practitioners, practice nurses and senior decision-makers, and desktop simulation exercises. We used the framework to analyse 89 publicly-available jurisdictional plans at similar managerial levels in the five countries. The framework identifies four functional domains: clinical care for influenza and other needs, public health responsibilities, the internal environment and the macro-environment of general practice. No plan addressed all four domains. Most plans either ignored or were sketchy about non-influenza clinical needs, and about the contribution of general practice to public health beyond surveillance. Collaborations between general practices were addressed in few plans, and inter-relationships with the broader health system, even less frequently. CONCLUSIONS: This is the first study to provide a framework to guide general practice planning for pandemic influenza. The framework helped identify critical shortcomings in available plans. Engaging general practice effectively in planning is challenging, particularly where governance structures for primary health care are weak. We identify implications for practice and for research. PMID- 18509539 TI - A reappraisal of azhdarchid pterosaur functional morphology and paleoecology. AB - Azhdarchid pterosaurs were among the most widespread and successful of pterosaur clades, but their paleoecology remains controversial. Morphological features common to all azhdarchids include a long, shallow rostrum; elongate, cylindrical cervical vertebrae that formed a long and unusually inflexible neck; and proportionally short wings with an abbreviated fourth phalanx. While azhdarchids have been imagined as vulture-like scavengers, sediment probers, swimmers, waders, aerial predators, or stork-like generalists, most recent authors have regarded them as skim-feeders, trawling their lower jaws through water during flight and seizing aquatic prey from the water's surface. Although apparently widely accepted, the skim-feeding model lacks critical support from anatomy and functional morphology. Azhdarchids lack the many cranial specialisations exhibited by extant skim-feeding birds, most notably the laterally compressed lower jaw and shock absorbing apparatus required for this feeding style. Well preserved azhdarchid skulls are rare, but their rostra and lower jaws appear to have been sub-triangular in cross-section, and thus dissimilar to those of skim feeders and sediment probers. Taphonomic data indicates that azhdarchids predominately inhabited inland settings, and azhdarchid morphology indicates that they were poorly suited for all proposed lifestyles bar wading and terrestrial foraging. However, azhdarchid footprints show that their feet were relatively small, padded and slender, and thus not well suited for wading. We argue that azhdarchids were stork- or ground hornbill-like generalists, foraging in diverse environments for small animals and carrion. Proficient terrestrial abilities and a relatively inflexible neck are in agreement with this interpretation. PMID- 18509540 TI - Effective detection of human leukocyte antigen risk alleles in celiac disease using tag single nucleotide polymorphisms. AB - BACKGROUND: The HLA genes, located in the MHC region on chromosome 6p21.3, play an important role in many autoimmune disorders, such as celiac disease (CD), type 1 diabetes (T1D), rheumatoid arthritis, multiple sclerosis, psoriasis and others. Known HLA variants that confer risk to CD, for example, include DQA1*05/DQB1*02 (DQ2.5) and DQA1*03/DQB1*0302 (DQ8). To diagnose the majority of CD patients and to study disease susceptibility and progression, typing these strongly associated HLA risk factors is of utmost importance. However, current genotyping methods for HLA risk factors involve many reactions, and are complicated and expensive. We sought a simple experimental approach using tagging SNPs that predict the CD associated HLA risk factors. METHODOLOGY: Our tagging approach exploits linkage disequilibrium between single nucleotide polymorphism (SNPs) and the CD associated HLA risk factors DQ2.5 and DQ8 that indicate direct risk, and DQA1*0201/DQB1*0202 (DQ2.2) and DQA1*0505/DQB1*0301 (DQ7) that attribute to the risk of DQ2.5 to CD. To evaluate the predictive power of this approach, we performed an empirical comparison of the predicted DQ types, based on these six tag SNPs, with those executed with current validated laboratory typing methods of the HLA-DQA1 and -DQB1 genes in three large cohorts. The results were validated in three European celiac populations. CONCLUSION: Using this method, only six SNPs were needed to predict the risk types carried by >95% of CD patients. We determined that for this tagging approach the sensitivity was >0.991, specificity >0.996 and the predictive value >0.948. Our results show that this tag SNP method is very accurate and provides an excellent basis for population screening for CD. This method is broadly applicable in European populations. PMID- 18509541 TI - Blood parasites in owls with conservation implications for the Spotted Owl (Strix occidentalis). AB - The three subspecies of Spotted Owl (Northern, Strix occidentalis caurina; California, S. o. occidentalis; and Mexican, S. o. lucida) are all threatened by habitat loss and range expansion of the Barred Owl (S. varia). An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp.) or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not been documented in the Spotted Owl. We screened 111 Spotted Owls, 44 Barred Owls, and 387 owls of nine other species for haemosporidian parasites (Leucocytozoon, Plasmodium, and Haemoproteus spp.). California Spotted Owls had the greatest number of simultaneous multi-species infections (44%). Additionally, sequencing results revealed that the Northern and California Spotted Owl subspecies together had the highest number of Leucocytozoon parasite lineages (n = 17) and unique lineages (n = 12). This high level of sequence diversity is significant because only one Leucocytozoon species (L. danilewskyi) has been accepted as valid among all owls, suggesting that L. danilewskyi is a cryptic species. Furthermore, a Plasmodium parasite was documented in a Northern Spotted Owl for the first time. West Coast Barred Owls had a lower prevalence of infection (15%) when compared to sympatric Spotted Owls (S. o. caurina 52%, S. o. occidentalis 79%) and Barred Owls from the historic range (61%). Consequently, Barred Owls on the West Coast may have a competitive advantage over the potentially immune compromised Spotted Owls. PMID- 18509542 TI - Structural dynamic of a self-assembling peptide d-EAK16 made of only D-amino acids. AB - We here report systematic study of structural dynamics of a 16-residue self assembling peptide d-EAK16 made of only D-amino acids. We compare these results with its chiral counterpart L-form, l-EAK16. Circular dichroism was used to follow the structural dynamics under various temperature and pH conditions. At 25 degrees C the d-EAK16 peptide displayed a typical beta-sheet spectrum. Upon increasing the temperature above 70 degrees C, there was a spectrum shift as the 218 nm valley widens toward 210 nm. Above 80 degrees C, the d-EAK16 peptide transformed into a typical alpha-helix CD spectrum without going through a detectable random-coil intermediate. When increasing the temperature from 4 degrees C to 110 degrees C then cooling back from 110 degrees C to 4 degrees C, there was a hysteresis: the secondary structure from beta-sheet to alpha-helix and then from alpha-helix to beta-sheet occurred. d-EAK16 formed an alpha-helical conformation at pH0.76 and pH12 but formed a beta-sheet at neutral pH. The effects of various pH conditions, ionic strength and denaturing agents were also noted. Since D-form peptides are resistant to natural enzyme degradation, such drastic structural changes may be exploited for fabricating molecular sensors to detect minute environmental changes. This provides insight into the behaviors of self-assembling peptides made of D-amino acids and points the way to designing new peptide materials for biomedical engineering and nanobiotechnology. PMID- 18509543 TI - Pentamidine dosage: a base/salt confusion. AB - Pentamidine has a long history in the treatment of human African trypanosomiasis (HAT) and leishmaniasis. Early guidelines on the dosage of pentamidine were based on the base-moiety of the two different formulations available. Confusion on the dosage of pentamidine arose from a different labelling of the two available products, either based on the salt or base moiety available in the preparation. We provide an overview of the various guidelines concerning HAT and leishmaniasis over the past decades and show the confusion in the calculation of the dosage of pentamidine in these guidelines and the subsequent published reports on clinical trials and reviews. At present, only pentamidine isethionate is available, but the advised dosage for HAT and leishmaniasis is (historically) based on the amount of pentamidine base. In the treatment of leishmaniasis this is probably resulting in a subtherapeutic treatment. There is thus a need for a new, more transparent and concise guideline concerning the dosage of pentamidine, at least in the treatment of HAT and leishmaniasis. PMID- 18509544 TI - A revision of Malagasy species of Anochetus mayr and Odontomachus latreille (Hymenoptera: Formicidae). AB - Species inventories are essential for documenting global diversity and generating necessary material for taxonomic study and conservation planning. However, for inventories to be immediately relevant, the taxonomic process must reduce the time to describe and identify specimens. To address these concerns for the inventory of arthropods across the Malagasy region, we present here a collaborative approach to taxonomy where collectors, morphologists and DNA barcoders using cytochrome c oxidase 1 (CO1) participate collectively in a team driven taxonomic process. We evaluate the role of DNA barcoding as a tool to accelerate species identification and description. This revision is primarily based on arthropod surveys throughout the Malagasy region from 1992 to 2006. The revision is based on morphological and CO1 DNA barcode analysis of 500 individuals. In the region, five species of Anochetus (A. boltonisp. nov., A. goodmanisp. nov., A. grandidieri, and A. madagascarensis from Madagascar, and A. pattersonisp. nov. from Seychelles) and three species of Odontomachus (O. coquereli, O. troglodytes and O. simillimus) are recognized. DNA barcoding (using cytochrome c oxidase 1 (CO1)) facilitated caste association and type designation, and highlighted population structure associated with reproductive strategy, biogeographic and evolutionary patterns for future exploration. This study provides an example of collaborative taxonomy, where morphology is combined with DNA barcoding. We demonstrate that CO1 DNA barcoding is a practical tool that allows formalized alpha-taxonomy at a speed, detail, precision, and scale unattainable by employing morphology alone. PMID- 18509545 TI - DNA methylation causes predominant maternal controls of plant embryo growth. AB - The parental conflict hypothesis predicts that the mother inhibits embryo growth counteracting growth enhancement by the father. In plants the DNA methyltransferase MET1 is a central regulator of parentally imprinted genes that affect seed growth. However the relation between the role of MET1 in imprinting and its control of seed size has remained unclear. Here we combine cytological, genetic and statistical analyses to study the effect of MET1 on seed growth. We show that the loss of MET1 during male gametogenesis causes a reduction of seed size, presumably linked to silencing of the paternal allele of growth enhancers in the endosperm, which nurtures the embryo. However, we find no evidence for a similar role of MET1 during female gametogenesis. Rather, the reduction of MET1 dosage in the maternal somatic tissues causes seed size increase. MET1 inhibits seed growth by restricting cell division and elongation in the maternal integuments that surround the seed. Our data demonstrate new controls of seed growth linked to the mode of reproduction typical of flowering plants. We conclude that the regulation of embryo growth by MET1 results from a combination of predominant maternal controls, and that DNA methylation maintained by MET1 does not orchestrate a parental conflict. PMID- 18509546 TI - Electromagnetic noise inhibits radiofrequency radiation-induced DNA damage and reactive oxygen species increase in human lens epithelial cells. AB - PURPOSE: The goal of this study was to investigate whether superposing of electromagnetic noise could block or attenuate DNA damage and intracellular reactive oxygen species (ROS) increase of cultured human lens epithelial cells (HLECs) induced by acute exposure to 1.8 GHz radiofrequency field (RF) of the Global System for Mobile Communications (GSM). METHODS: An sXc-1800 RF exposure system was used to produce a GSM signal at 1.8 GHz (217 Hz amplitude-modulated) with the specific absorption rate (SAR) of 1, 2, 3, and 4 W/kg. After 2 h of intermittent exposure, the ROS level was assessed by the fluorescent probe, 2',7' dichlorodihydrofluorescein diacetate (DCFH-DA). DNA damage to HLECs was examined by alkaline comet assay and the phosphorylated form of histone variant H2AX (gammaH2AX) foci formation assay. RESULTS: After exposure to 1.8 GHz RF for 2 h, HLECs exhibited significant intracellular ROS increase in the 2, 3, and 4 W/kg groups. RF radiation at the SAR of 3 W/kg and 4 W/kg could induce significant DNA damage, examined by alkaline comet assay, which was used to detect mainly single strand breaks (SSBs), while no statistical difference in double strand breaks (DSBs), evaluated by gammaH2AX foci, was found between RF exposure (SAR: 3 and 4 W/kg) and sham exposure groups. When RF was superposed with 2 muT electromagnetic noise could block RF-induced ROS increase and DNA damage. CONCLUSIONS: DNA damage induced by 1.8 GHz radiofrequency field for 2 h, which was mainly SSBs, may be associated with the increased ROS production. Electromagnetic noise could block RF-induced ROS formation and DNA damage. PMID- 18509547 TI - Age-dependent association of gamma-crystallins with aged alpha-crystallins from old bovine lens. AB - PURPOSE: Previous theoretical and experimental studies have predicted that the loss of weak protein interactions between alpha- and gamma-crystallins could result in a decrease in the transparent properties of the aging lens. METHODS: alpha-Crystallins were prepared from the nucleus of old bovine lens, and gamma crystallins were prepared from whole fetal bovine lens or from the nucleus of old bovine lens. The possible interactions of old alpha-crystallins with either old gamma-crystallins or fetal gamma-crystallins were quantitated at equilibrium using microequilibrium dialysis. The amount of each gamma-crystallin species in the "full" versus "empty" chambers was determined by reverse phase chromatography to obtain a binding ratio (full/empty). RESULTS: A binding ratio greater than 1.00 is indicative of a alpha-crystallin/gamma-crystallin interaction. Within experimental error (+/-2X standard deviation), there were no interactions between aged gamma-crystallins with aged alpha-crystallins while there were significant interactions between some of the fetal gamma-crystallins with aged alpha crystallins. CONCLUSIONS: In the aged bovine lens, when transparency is known to decrease, there is no detectable interaction of gamma-crystallins with alpha crystallins as measured by microequilibrium dialysis. The results are consistent with the hypothesis that short-range, weak, attractive interactions between alpha and gamma-crystallins are necessary for maximum transparency of the lens. PMID- 18509548 TI - The protective role of transferrin in Muller glial cells after iron-induced toxicity. AB - PURPOSE: Transferrin (Tf) expression is enhanced by aging and inflammation in humans. We investigated the role of transferrin in glial protection. METHODS: We generated transgenic mice (Tg) carrying the complete human transferrin gene on a C57Bl/6J genetic background. We studied human (hTf) and mouse (mTf) transferrin localization in Tg and wild-type (WT) C57Bl/6J mice using immunochemistry with specific antibodies. Muller glial (MG) cells were cultured from explants and characterized using cellular retinaldehyde binding protein (CRALBP) and vimentin antibodies. They were further subcultured for study. We incubated cells with FeCl(3)-nitrilotriacetate to test for the iron-induced stress response; viability was determined by direct counting and measurement of lactate dehydrogenase (LDH) activity. Tf expression was determined by reverse transcriptase-quantitative PCR with human- or mouse-specific probes. hTf and mTf in the medium were assayed by ELISA or radioimmunoassay (RIA), respectively. RESULTS: mTf was mainly localized in retinal pigment epithelium and ganglion cell layers in retina sections of both mouse lines. hTf was abundant in MG cells. The distribution of mTf and hTf mRNA was consistent with these findings. mTf and hTf were secreted into the medium of MG cell primary cultures. Cells from Tg mice secreted hTf at a particularly high level. However, both WT and Tg cell cultures lose their ability to secrete Tf after a few passages. Tg MG cells secreting hTf were more resistant to iron induced stress toxicity than those no longer secreted hTf. Similarly, exogenous human apo-Tf, but not human holo-Tf, conferred resistance to iron-induced stress on MG cells from WT mice. CONCLUSIONS: hTf localization in MG cells from Tg mice was reminiscent of that reported for aged human retina and age-related macular degeneration, both conditions associated with iron deposition. The role of hTf in protection against toxicity in Tg MG cells probably involves an adaptive mechanism developed in neural retina to control iron-induced stress. PMID- 18509549 TI - Psf2 plays important roles in normal eye development in Xenopus laevis. AB - PURPOSE: Psf2 (partner of Sld5 2) represents a member of the GINS (go, ichi, ni, san) heterotetramer [1] and functions in DNA replication as a "sliding clamp." Previous in situ hybridization analyses revealed that Psf2 is expressed during embryonic development in a tissue-specific manner, including the optic cup (retina) and the lens [2]. This article provides an analysis of Psf2 function during eye development in Xenopus laevis. METHODS: A morpholino targeted to Psf2 mRNA was designed to knockdown Psf2 translation and was injected into specific embryonic cells during early cleavage stages in the frog, Xenopus laevis. Injected embryos were assayed for specific defects in morphology, cell proliferation, and apoptosis. Synthetic Psf2 RNA was also co-injected with the morpholino to rescue morpholino-mediated developmental defects. It is well known that reciprocal inductive interactions control the development of the optic cup and lens. Therefore, control- and morpholino-injected embryos were used for reciprocal transplantation experiments to distinguish the intrinsic role of Psf2 in the development of the optic cup (retina) versus the lens. RESULTS: Morpholino mediated knockdown of Psf2 expression resulted in dosage-dependent phenotypes, which included microphthalmia, incomplete closure of the ventral retinal fissure, and retinal and lens dysgenesis. Defects were also observed in other embryonic tissues that normally express Psf2 including the pharyngeal arches and the otic vesicle, although other tissues that express Psf2 were not found to be grossly defective. Eye defects could be rescued by co-injection of synthetic Psf2 RNA. Examination of cell proliferation via an antibody against phospho-histone H3 S10P revealed no significant differences in the retina and lens following Psf2 knockdown. However, there was a significant increase in the level of apoptosis in retinal as well as forebrain tissues, as revealed by TUNEL (terminal deoxynucleotide transferase dUTP nick end labeling) assay. CONCLUSIONS: The results demonstrate intrinsic roles for Psf2 in both retinal and to a lesser extent, lens tissues. Observed lens defects can mainly be attributed to deficiencies in retinal development and consequently the late phase of lens induction, which involves instructive cues from the optic cup. Developmental defects were not observed in all tissues that express Psf2, which could be related to differences in the translation of Psf2 or redundant effects of related factors such as proliferating cell nuclear antigen (PCNA). PMID- 18509550 TI - Karyotype changes in cultured human corneal endothelial cells. AB - PURPOSE: To examine karyotype changes in cultured human corneal endothelial cells (HCECs). METHODS: HCECs with Descemet's membrane were removed from 20 donors of various ages (range, 2-77 years; average, 43.7+/-26.4 years) and cultured on dishes coated with extracellular matrix produced by bovine corneal endothelial cells (BCECs). Karyotype changes were examined by G-band karyotyping of HCECs at the third passage from 12 donors and the fifth passage from 16 donors. The number of chromosomes was analyzed in 40-50 cells from the third and fifth passages of each HCEC preparation. A detailed karyotype analysis of 10-16 cells from the third and fifth passages of each HCEC preparation was performed. The frequency of aneuploid cells per case (the number of abnormal cells divided by the total number of cells examined at metaphase) was tested for correlation with age by Spearman's correlation analysis. RESULTS: At the third passage, five cases (41.7%) showed an almost normal karyotype, and five cases (41.7%) showed sex chromosome loss. One case (8.3%) showed chromosome 21 trisomy. At the fifth passage, five cases (31.3%) showed an almost normal karyotype, and four cases (25%) showed sex chromosome loss. Three cases (18.8%) showed chromosome 8 trisomy, and one case (6.3%) showed chromosome 21 trisomy. Donor age and the frequency of aneuploidy had a statistically significant correlation at the fifth passage (R=0.653, p=0.042). CONCLUSIONS: Donor age and the frequency of aneuploidy have a positive correlation in cultured HCECs at the fifth passage. Therefore, HCECs for clinical therapies should be obtained from donors as young as possible. Karyotyping cultured HCECs is crucial before clinical application. PMID- 18509552 TI - Mutations in the TOPORS gene cause 1% of autosomal dominant retinitis pigmentosa. AB - PURPOSE: The purpose of this project was to determine if mutations, including large insertions or deletions, in the recently identified RP31 gene topoisomerase I-binding arginine-serine rich (RS) protein (TOPORS), cause an appreciable fraction of autosomal dominant retinitis pigmentosa (adRP). METHODS: An adRP cohort of 215 families was used to determine the frequency of TOPORS mutations. We looked for mutations in TOPORS by testing 89 probands from the cohort without mutations in other known adRP genes. Mutation detection was performed by fluorescent capillary sequencing and by multiplex ligation probe amplification. RESULTS: Two different TOPORS mutations, p.Glu808X and p.Arg857GlyfsX9, were each identified in one proband. Patients with these mutations exhibited clinical signs typical of advanced adRP. No large deletions or insertions of TOPORS were identified in our study. CONCLUSIONS: Point mutations and small insertions or deletions in TOPORS cause approximately 1% of adRP. Large deletions or insertions of TOPORS are not an appreciable cause of adRP. Contrary to previous reports, no distinct clinical phenotype was seen in these patients. PMID- 18509551 TI - The Tg(ccnb1:EGFP) transgenic zebrafish line labels proliferating cells during retinal development and regeneration. AB - PURPOSE: To create the Tg(ccnb1:EGFP)(nt18) zebrafish line that spatially and temporally labels retinal progenitor cells with enhanced green fluorescent protein (EGFP) during zebrafish retinal development and regeneration. METHODS: We cloned the 1.5 kb promoter region of the zebrafish cyclin B1 (ccnb1) gene upstream of the EGFP gene in the Tol2 vector, which was used to generate the stable Tg(ccnb1:EGFP)(nt18) transgenic zebrafish line. Immunohistochemistry and in situ hybridization techniques verified that the ccnb1:EGFP transgene was expressed in retinal progenitor cells during retinal development, in the undamaged adult retina, and in the regenerating adult retina. RESULTS: At 36 h post-fertilization, both the enhanced green fluorescent protein (EGFP) and proliferating cell nuclear antigen (PCNA) expressions were observed throughout the developing transgenic retina, but they became restricted to the circumferential marginal zone by five days post-fertilization. In situ hybridization confirmed that this EGFP expression matched the cyclin B1 mRNA expression pattern. In comparison to the Tg(1016a1tubulin:EGFP) transgenic line that expresses EGFP in neuronal progenitor cells, the Tg(ccnb1:EGFP)(nt18) line more faithfully follows the rise and fall of PCNA expression through the developing retina and brain. In the adult retina, there are three cell types that continue to proliferate, the Muller glia in the inner nuclear layer, the rod precursor cells in the outer nuclear layer, and the stem cells in the circumferential marginal zone. In the Tg(ccnb1:EGFP)(nt18) retina, EGFP coexpressed with PCNA in all three of these proliferating cell types. Exposing the adult retina to constant intense light destroys the rod and cone photoreceptors and induces an increase in the number of proliferating Muller glia, which produces actively dividing neuronal progenitor cells that migrate to the outer nuclear layer (ONL) and replenish the lost photoreceptors. Following constant light damage, Tg(ccnb1:EGFP)(nt18) zebrafish expressed EGFP in both the proliferating Muller glia and the migrating neuronal progenitor cells. CONCLUSIONS: The spatial and temporal patterning of EGFP expression in the Tg(ccnb1:EGFP)(nt18) line directly reflects the known locations of proliferating cells in the zebrafish retina, making it a useful marker to study the transient nature of neuronal progenitor cells during the development and regeneration of the zebrafish retina. PMID- 18509553 TI - Linking health promotion with physiotherapy for low back pain: a review. AB - OBJECTIVE: The objectives of this paper are: (i) to present the results of a descriptive literature review highlighting conceptual and practical links between the fields of physiotherapy and health promotion, and (ii) to provide recommendations based on this review of the literature in order to contribute towards the improvement of physiotherapists' interventions with people presenting low back pain. METHODS: A literature review of publications in the fields of health promotion, public health, physiotherapy and rehabilitation. The concepts of health and empowerment are discussed. Health promotion strategies used in the field of physiotherapy are also reported. RESULTS: The results of the literature review indicate that conceptualizations of health differ between the fields of health promotion and physiotherapy, although there are some common points. Empowerment, a central concept in health promotion, is probably not facilitated in physiotherapy interventions based on the biomedical model. Health education is the most used health promotion strategy in physiotherapy practice. Recommendations are put forward. CONCLUSION: In the future, further efforts should be made towards linking the principles and practices of health promotion with physiotherapy. This may help improve physiotherapists' interventions with people presenting low back pain. PMID- 18509554 TI - Comprehensive rehabilitation at Beitostolen Healthsports Centre: influence on mental and physical functioning. AB - OBJECTIVE: To characterize the mental and physical functioning of subjects admitted to Beitostolen Healthsports Centre, and evaluate whether functioning was improved during rehabilitation. MATERIAL AND METHODS: A total of 228 subjects admitted for rehabilitation completed the Medical Outcome Study Short Form 12 (SF 12) on admission and on discharge after approximately 4 weeks. A total of 174 subjects also completed SF-12 3 months after discharge. The physical and mental sum scores and the scores for the 8 subscales of SF-12 were calculated. In a stepwise backward multiple regression model, demographic and medical factors influencing improvement during rehabilitation were explored. RESULTS: Physical, as well as mental, functioning was reduced on admission. Bodily pain and emotional factors represented the most dominant components. Both physical and mental functioning improved during the stay, and the improvement in physical functioning remained stable 3 months after discharge. The improvement in physical functioning was greatest for those subjects living in urban areas and having a painful musculoskeletal diagnosis. The improvement in mental functioning was greatest for females. CONCLUSION: This approach to rehabilitation appears to be effective for improving physical functioning in the relevant group of individuals. However, carefully controlled studies with longer follow-up periods are required. PMID- 18509555 TI - Assessment of motion in the cervico-thoracic spine in patients with subacute whiplash-associated disorders. AB - OBJECTIVE: To investigate the distribution of segmental flexion mobility in the cervico-thoracic spine of men and women with whiplash-associated disorders. The study also assesses the relationship between cervical mobility and segmental flexion mobility, and whether hypomobility in C7-T1 is associated with neck pain and weakness in the hands. Finally, the study investigates the impact of fear of movement/(re)injury and pain intensity on cervical mobility. METHOD: The data in this longitudinal study was obtained from a previous trial on 47 patients. Assessments were performed at baseline and 3 and 9 months following randomization using a cervical range of motion instrument, the cervico-thoracic ratio, a Grippit, a visual analogue scale, a Painmatcher, and the Tampa Scale for Kinesiophobia. RESULTS: Women exhibited hypomobility in C7-T1. There was no significant association between the 2 cervical measurement techniques. The correlation between flexion mobility in C7-T1 and grip strength was weak but significant (r=-0.3) (p<0.05). There was also a weak but significant negative correlation between total cervical mobility and neck pain intensity. No significant correlation was found between fear of movement/(re)injury and cervical mobility. CONCLUSION: Women with whiplash-associated disorders seem to exhibit flexion hypomobility in C7-T1. The flexion mobility in C7-T1 was weakly, but significantly, correlated with grip strength, but was not shown to predict neck pain. Neck pain may give rise to restricted range of motion. PMID- 18509556 TI - Altered interpretation of neck proprioceptive signals in persons with subclinical recurrent neck pain. AB - OBJECTIVE: To evaluate whether subjects with subclinical recurrent neck pain have an altered interpretation of neck proprioceptive signals. DESIGN: A comparative group design. SUBJECTS: Twelve subjects with recurrent neck pain and 12 control subjects. METHODS: The shoulder, head, trunk and whole-body position were measured under the following conditions: active and passive elevation and depression of the right shoulder and vibration of the trapezius muscle. RESULTS: During passive shoulder movements both groups moved their head in the opposite direction (p<0.05). During passive elevation subjects with recurrent neck pain showed a significant over-appraisal of shoulder position. Both groups responded similarly to active movements. Subjects with recurrent neck pain and control subjects showed similar trunk and head-to-trunk movements during shoulder movements. Subjects with recurrent neck pain, however, made larger movements compared with healthy subjects (p<0.05). During trapezius muscle vibration similar whole-body movements were found in both groups. CONCLUSION: These results show a modified interpretation of neck proprioceptive signals in subjects with recurrent neck pain and may reflect an offset in the egocentric reference frame or a decreased capacity to switch between reference frames. Better insight into these mechanisms might lead to better evaluation and treatment of subjects with recurrent neck pain and to a reduction in recurrent episodes. PMID- 18509557 TI - Manual therapy in addition to physiotherapy does not improve clinical or economic outcomes after ankle fracture. AB - OBJECTIVE: The primary aim of this study was to determine the effectiveness and cost-effectiveness of adding manual therapy to a physiotherapy programme for ankle fracture. DESIGN: Assessor-blinded randomized controlled trial. PARTICIPANTS: Ninety-four adults were recruited within one week of cast removal for isolated ankle fracture. Inclusion criteria were: they were able to weight bear as tolerated or partial weight-bear, were referred for physiotherapy, and experienced pain. Ninety-one participants completed the study. METHODS: Participants were randomly allocated to receive manual therapy (anterior posterior joint mobilization over the talus) plus a standard physiotherapy programme (experimental), or the standard physiotherapy programme only (control). They were assessed by a blinded assessor at baseline, and at 4, 12 and 24 weeks. The main outcomes were activity limitation and quality of life. Information on costs and healthcare utilization was collected every 4 weeks up to 24 weeks. RESULTS: There were no clinically worthwhile differences in activity limitation or quality of life between groups at any time-point. There was also no between group difference in quality-adjusted life-years, but the experimental group incurred higher out-of-pocket costs (mean between-group difference = AU$200, 95% confidence interval 26-432). CONCLUSION: When provided in addition to a physiotherapy programme, manual therapy did not enhance outcome in adults after ankle fracture. PMID- 18509558 TI - Worker's compensation and return-to-work following orthopaedic injury to extremities. AB - OBJECTIVE: To explore the influence of workers' compensation coverage on injury patterns and return-to-work outcome in orthopaedic injuries of the extremities. DESIGN: Prospective and follow-up study. SUBJECTS: A total of 154 subjects were recruited and 37.7% of the injuries were covered by workers' compensation. METHODS: Baseline data were obtained by questionnaire and chart review. The main outcome variable was time of first return to work. Subjects were followed up at 1, 3 and 6 months after initial interview. Cox proportional hazards model was used to analyse associations between potential predictors and return-to-work. RESULTS: The workers' compensation group were most likely to be men employed in labour occupations and were most likely to sustain crushing injuries, or injuries from being struck by objects. The 6-month return-to-work rates for the workers' compensation and non-workers' compensation groups were 70.7% and 71.9%. Early return-to-work was associated with more years in higher education, and increased self-efficacy in both groups. Moreover, age older than 45 years and hospitalization less than 14 days were associated with early return-to-work in the non-workers' compensation group. CONCLUSION: The injury patterns of workers' compensation and non-workers' compensation groups differed, but the likelihood of return-to-work at 6-month follow-up was similar. More years of education and self efficacy were positive predictors of return-to-work. PMID- 18509559 TI - A single home visit by an occupational therapist reduces the risk of falling after hip fracture in elderly women: a quasi-randomized controlled trial. AB - OBJECTIVE: To assess the effectiveness of a single home visit by an occupational therapist in the reduction of fall risk after hip fracture in elderly women. DESIGN: Quasi-randomized controlled trial. PARTICIPANTS: Ninety-five women aged 60 years or older, living in the community, who sustained a fall-related hip fracture. METHODS: The women were allocated alternately to intervention or control groups. All the women underwent a multidisciplinary programme targeted at fall prevention during in-patient rehabilitation. Additionally, the intervention group received a home visit by an occupational therapist a median of 20 days after discharge. Falls were recorded at a 6-month follow-up. RESULTS: Thirteen of the 50 women in the control group sustained 20 falls during 9231 days, whereas 6 of the 45 women in the intervention group sustained 9 falls during 8970 days. After adjustment for observation periods, Barthel Index scores, and body height, a significantly lower proportion of fallers was found in the intervention group: the odds ratio was 0.275 (95% confidence interval 0.081-0.937, p=0.039). CONCLUSION: A single home visit by an occupational therapist after discharge from a rehabilitation hospital significantly reduced the risk of falling in a sample of elderly women following hip fracture. PMID- 18509560 TI - Specific exercises reduce brace prescription in adolescent idiopathic scoliosis: a prospective controlled cohort study with worst-case analysis. AB - OBJECTIVE: To compare the effect of Scientific Exercises Approach to Scoliosis (SEAS) exercises with "usual care" rehabilitation programmes in terms of the avoidance of brace prescription and prevention of curve progression in adolescent idiopathic scoliosis. DESIGN: Prospective controlled cohort observational study. PATIENTS: Seventy-four consecutive outpatients with adolescent idiopathic scoliosis, mean 15 degrees (standard deviation 6) Cobb angle, 12.4 (standard deviation 2.2) years old, at risk of bracing who had not been treated previously. METHODS: Thirty-five patients were included in the SEAS exercises group and 39 in the usual physiotherapy group. The primary outcome included the number of braced patients, Cobb angle and the angle of trunk rotation. RESULTS: There were 6.1% braced patients in the SEAS exercises group vs 25.0% in the usual physiotherapy group. Failures of treatment in the worst-case analysis were 11.5% and 30.8%, respectively. In both cases the differences were statistically significant. Cobb angle improved in the SEAS exercises group, but worsened in the usual physiotherapy group. In the SEAS exercises group, 23.5% of patients improved and 11.8% worsened, while in the usual physiotherapy group 11.1% improved and 13.9% worsened. CONCLUSION: These data confirm the effectiveness of exercises in patients with scoliosis who are at high risk of progression. Compared with non adapted exercises, a specific and personalized treatment (SEAS) appears to be more effective. PMID- 18509561 TI - Relationship of glenohumeral elevation and 3-dimensional scapular kinematics with disability in patients with shoulder disorders. AB - OBJECTIVE: To determine variables among glenohumeral elevation and 3-dimensional scapular rotations that are related to shoulder function as assessed by the Disability of the Arm, Shoulder and Hand-Disability/Symptom (DASH-D/S) scale. DESIGN: Prospective, cross-sectional study. PATIENTS: Eighty-eight patients with shoulder pain and limited range of motion were included. METHODS: Each patient performed 2 full active range of motion activities, forward flexion and abduction, and 2 activities of daily living, combing hair and simulating washing the back. Glenohumeral elevation and scapular rotations were measured by the Polhemus Fastrak electromagnetic system. RESULTS: On multiple regression analysis, glenohumeral elevation in combing hair and scapular lateral rotation in both abduction and simulating washing the back were the best predictors of shoulder function and explained 39.7% of the variance of the DASH-D/S score. CONCLUSION: These findings support the classical rehabilitation of the shoulder based on glenohumeral elevation and suggest the importance of attention paid to scapular lateral rotation. PMID- 18509562 TI - Barriers to and facilitators of everyday physical activity in persons with a spinal cord injury after discharge from the rehabilitation centre. AB - OBJECTIVE: To determine the most important barriers to and facilitators of the level of everyday physical activity in persons with a spinal cord injury after discharge from the rehabilitation centre. DESIGN: Qualitative study with both cross-sectional and retrospective questions. SUBJECTS: Thirty-two persons with a spinal cord injury. METHODS: Semi-structured interview with questions concerning the current situation (>9 months after discharge) and retrospective questions concerning the period shortly after discharge (=3 months). The interview consisted of 10 topic categories assumed to have an impact on the level of everyday physical activity and covering the main parts of the International Classification of Functioning, Disability and Health (ICF) model. RESULTS: In the current situation, the most important barriers were problems with accessibility of stores and buildings, physical health problems and mental health problems. Shortly after discharge, the most important barriers were emotional distress, problems with self-care, and mental health problems. The most frequently mentioned facilitators were preparation in the rehabilitation centre with respect to daily activities and social activities and stimulation to be physically active. CONCLUSION: Persons with a spinal cord injury experience important barriers to physical activity, particularly on the ICF component Body Functions and Structure. PMID- 18509563 TI - Quantification of reaction forces during sitting pivot transfers performed by individuals with spinal cord injury. AB - OBJECTIVES: To quantify the reaction forces exerted under the hands, feet and buttocks when individuals with spinal cord injury performed sitting pivot transfers. DESIGN: Twelve men with paraplegia completed 3 transfers between seats of the same height (0.5 m high) and 3 transfers to a high target seat (0.6 m high). RESULTS: Greater mean and peak vertical reaction forces were always recorded under the hands compared with the feet (p<0.001) during the transfers. Mean vertical reaction forces were similar between the leading and trailing hands (p>0.088) for the 2 transfers studied. However, the mean vertical reaction force underneath the leading hand was greater when transferring between a seat of the same height compared with one of a higher height (p=0.021) and vice-versa for the trailing hand (p=0.0001). The peak vertical reaction force always occurred earlier (p<0.0001) and was greater underneath the trailing hand compared with the leading one (p<0.02), and reached its highest value when transferring to the high target seat (p=0.003). Peak and mean horizontal reaction forces were always higher underneath the trailing hand compared with the leading hand (p<0.001). CONCLUSION: These results provide evidence-based data to better understand transfers and strengthen clinical practice guidelines targeting the preservation of upper extremity integrity. PMID- 18509564 TI - Positive emotion following a stroke. AB - OBJECTIVE: Interest in exploring the positive emotional health of adults has increased. The current study investigated the positive emotion of adults with stroke at discharge from in-patient medical rehabilitation and 3 months post discharge. DESIGN: A longitudinal study in which information was collected during in-patient medical rehabilitation stay and 3 months post-discharge. SUBJECTS: The study included 856 persons with stroke aged 55 years or older. RESULTS: Mean age for the sample was 72.5 years, 78.7% of subjects were non-Hispanic white and 51.9% were women. Mean length of hospital stay was 20.2 days. More than one-third of patients reported higher positive emotion over the 3-month follow-up, while 29.7% reported lower positive emotion. In addition to discharge positive emotion, 4 factors, including depression, level of education and motor and cognition functional status score, significantly predicted lower depression at 3-month follow-up. CONCLUSION: A large percentage of adults reported high positive emotion in the initial months following a stroke. This finding adds to work on stroke recovery and indicates the emotional resilience of adults when faced with a health challenge. Understanding the role of positive emotion in persons living with stroke may provide insight into long-term recovery. PMID- 18509565 TI - Kinematics of the contralateral and ipsilateral shoulder: a possible relationship with post-stroke shoulder pain. AB - OBJECTIVE: Post-stroke shoulder pain is a common phenomenon in hemiplegia and impedes rehabilitation. The aim of this study was to identify a possible relationship between post-stroke shoulder pain, scapula resting position and shoulder motion. METHODS: Shoulder kinematics of 27 patients after stroke (17 men) were compared with 10 healthy age-matched control subjects. Using an electromagnetic tracking device, the kinematics of both the contralateral and ipsilateral (i.e. paretic and non-paretic) arm during active and passive abduction and forward flexion were measured and expressed in Euler angles. RESULTS: Scapular lateral rotation relative to the thorax was increased in patients with post-stroke shoulder pain compared with both patients without post stroke shoulder pain and control subjects at rest as well as during arm abduction and forward flexion. Additionally, glenohumeral elevation was decreased in patients with post-stroke shoulder pain during passive abduction. No differences were found regarding scapula position (displacement relative to the thorax). CONCLUSION: In patients with post-stroke shoulder pain a particular kinematical shoulder pattern was established, characterized by enhanced scapular lateral rotation and diminished glenohumeral mobility. PMID- 18509566 TI - Functional recovery differs between ischaemic and haemorrhagic stroke patients. AB - OBJECTIVE: To determine whether there is a difference between patients with a cerebral infarction and those with an intracerebral haemorrhage with respect to the development of independence in activities of daily living over the first year post-stroke. METHODS: Patients after first-ever stroke who were admitted to an inpatient rehabilitation programme were included. The study had a longitudinal design and measurements took place at admission, 8, 10, 12, 26 and 52 weeks post stroke. The relationship between the development over time of activities of daily living independence, measured by the Barthel Index, and type of stroke was analysed using Generalized Estimating Equations. RESULTS: A total of 229 patients with cerebral infarction and 45 with intracerebral haemorrhage were included. From 12 to 26 weeks post-stroke, patients with cerebral infarction showed a significantly faster recovery. The time window for recovery was more restricted for patients with intracerebral haemorrhage; a statistically significant increase in activities of daily living was found until 10 weeks post-stroke in patients with intracerebral haemorrhage, whereas patients with cerebral infarction showed statistically significant recovery until 26 weeks post-stroke. CONCLUSION: The differences in activities of daily living recovery between patients with cerebral infarction and those with intracerebral haemorrhage should be taken into consideration in rehabilitation management. PMID- 18509567 TI - Organizing human functioning and rehabilitation research into distinct scientific fields. PMID- 18509568 TI - Publishing in physical and rehabilitation medicine: a European point of view. PMID- 18509570 TI - Comparison of individual, group and combined intervention formats in a randomized controlled trial for facilitating goal attainment and improving psychosocial function following acquired brain injury. AB - OBJECTIVE: To compare individual, group and combined intervention formats for improving goal attainment and psychosocial function following acquired brain injury. DESIGN: Randomized controlled trial, waiting list controls. PARTICIPANTS: Thirty-five participants with a mean time of 5.29 years (standard deviation = 3.9) since acquired brain injury were randomly allocated into 6 groups involving an intervention or waiting list control condition for 1 of 3 intervention formats. METHODS: Interventions were 3 h/week for 8 weeks. Formats included: group-based support (n = 12), individual occupation-based support (n = 11), and a combined group and individual support intervention (n = 12). Participant outcomes were examined at pre-, post-, and 3-month follow-up assessment on the Canadian Occupational Performance Measure, Patient Competency Rating Scale, and Brain Injury Community Rehabilitation Outcome 39 Scales. RESULTS: Overall, the findings indicated that the individual intervention component appeared to contribute particularly to gains in performance in goal-specific areas. The combined intervention was associated with maintained gains in performance and satisfaction. However, gains in behavioural competency and psychological well being were more likely to occur after the group and individual interventions. CONCLUSION: These findings generally support the efficacy of brief intervention formats following acquired brain injury, although further research is needed to examine clients' suitability for particular interventions. PMID- 18509571 TI - The complexity of participation in daily life: a qualitative study of the experiences of persons with acquired brain injury. AB - OBJECTIVE: To describe and enhance the understanding of how adults with acquired brain injury experience participation in daily life. PATIENTS AND METHODS: Qualitative interviews with 11 persons of working age with acquired brain injuries were analysed using qualitative content analysis. RESULTS: The informants' experiences formed 5 categories: "Performing tasks"; "Making decisions and exerting influence"; "Being engaged in meaningful activities"; "Doing things for others"; and "Belonging". The categories that needed to be present for the informants to experience a feeling of participation varied according to their individual daily life situations. In addition, their experiences showed that a variety of conditions, related to each of the 5 categories, influenced their participation. Individuals adopted a variety of strategies to enhance their experience of participation. CONCLUSION: The meaning of participation and the conditions and strategies influencing participation are complex. Many of the categories identified for participation can be understood only through subjective experience and cannot be captured by professionals' observation of the performance of activities. These results emphasize the importance of considering clients' unique experiences of participation when designing individually tailored rehabilitation programmes intended to enhance participation. PMID- 18509572 TI - Impact of internal capsule lesions on outcome of motor hand function at one year post-stroke. AB - OBJECTIVE: To investigate the association between damage to different levels of the corticofugal tract and long-term hand motor recovery. DESIGN: Prospective cohort study. PATIENTS: Seventy-five first-ever middle cerebral artery stroke survivors. METHODS: Hand motor function was assessed with the Fugl-Meyer Motor Assessment Scale at one year post-stroke. Localization of the corticofugal tract was assessed on axial magnetic resonance imaging slices using a corticofugal mask defining involvement of the motor cortex, corona radiata or internal capsule and combinations of these structures, adjusted for lesion volume. RESULTS: Longitudinal involvement of all 3 levels of the corticofugal tract and partial involvement of the internal capsule were associated with a significant probability of poor motor hand function recovery. The probability of regaining hand function ranged from 54% if the corticofugal tract was only partly affected to 13% if both motor cortex and internal capsule were affected. CONCLUSION: At one year post-stroke, lesions of the internal capsule were associated with a significantly lower probability of return of isolated hand motor function than lesions of the cortex, subcortex and corona radiata. Since recovery of isolated hand and finger movements is important for regaining a functional upper limb in everyday living, these patients should be identified early post-stroke in order to evaluate specific hand function training. PMID- 18509573 TI - Mapping the Stroke Impact Scale (SIS-16) to the International Classification of Functioning, Disability and Health. AB - OBJECTIVE: To demonstrate how the International Classification of Functioning, Disability and Health (ICF) can be used to create coded functional status indicators specific for stroke from a simple stroke-specific functional index, the Stroke Impact Scale-16 (SIS-16). SUBJECTS: Nineteen professionals for the mapping portion and 8 persons with stroke for the cognitive debriefing portion. METHODS: Participants were asked to identify appropriate codes for the corresponding items of the SIS-16 following a structured protocol for mapping measures to the ICF. A Delphi technique was used in order to reach consensus for as many items as possible. In addition, cognitive debriefing was conducted with persons with stroke. RESULTS: A total of 13 items had Functional Status Indicators endorsed (8 items at the 4 digit level and 5 items at the 3 digit level). There were 3 items that did not reach consensus. The cognitive debriefing sessions demonstrated the differences in interpretation from the persons with stroke and the intentions by the developers. CONCLUSION: This study has shown how the ICF can capture most items from functional status measures, such as the SIS 16. Furthermore, the items can be used to map onto a standard coding framework, illustrating the potential for increased use of Functional Status Indicators. PMID- 18509574 TI - Isotonic and isometric contractions exert the same amount of corticomotor system excitability in healthy subjects and patients after stroke. AB - OBJECTIVE: Active, voluntary training of the centrally paretic upper limb is crucial for functional recovery after brain damage. The aim of this study was to determine whether the type of voluntary contraction has a differential influence on corticomotor system excitability in healthy subjects and patients after stroke. DESIGN: Experimental cross-sectional study. SUBJECTS: Fifteen healthy volunteers and 15 patients after stroke. METHODS: Participants performed dynamic isotonic and isometric voluntary wrist extensions with the non-dominant or the paretic hand, respectively, with force levels of 10%, 20% and 30% of the maximum voluntary surface electromyogram. Excitability was measured by comparing the amplitude of motor evoked potentials elicited by transcranial magnetic stimulation. RESULTS: The type of contraction did not have any effect on the amplitude of motor evoked potentials, either in healthy subjects or in patients after stroke. CONCLUSION: Dynamic isometric and isotonic voluntary contractions seem to have the same effect on the excitability of the corticomotor system. PMID- 18509575 TI - Social, intimate and sexual relationships of adolescents with cerebral palsy compared with able-bodied age-mates. AB - OBJECTIVE: To describe the social, intimate and sexual relationships of Dutch adolescents with cerebral palsy compared with their able-bodied age mates. DESIGN: Cross-sectional study. SUBJECTS: A total of 103 adolescents with cerebral palsy without severe learning problems aged 16-20 years. METHODS: We used a structured interview and questionnaires to assess subject characteristics such as age, type of cerebral palsy, gross motor function and level of education. Main outcome measures on social, intimate and sexual relationships are the Life-Habits questionnaire, the Vineland Adaptive Behaviour Scale, and a structured interview developed for Dutch studies in able-bodied persons and persons with spina bifida. Experienced competence was assessed with the Dutch version of the Self-Perception Profile of Adolescents and the Physical Disability Sexual and Body Esteem Scale. These data were compared with matching reference data, mainly from able-bodied (Dutch) adolescents. RESULTS: Approximately 30% of the subjects functioned socially below their age level. Adolescents with cerebral palsy find it difficult to develop intimate relationships and they have less sexual experience than their able-bodied age mates. CONCLUSION: Although adolescents with cerebral palsy do have social relationships, it is difficult for them to develop intimate relationships. They perceive various barriers, but seem to have a positive self perception. PMID- 18509576 TI - Patients with acute spinal cord injury benefit from normocapnic hyperpnoea training. AB - BACKGROUND: Functional loss of respiratory muscles in persons with spinal cord injury leads to impaired pulmonary function and respiratory complications. In addition, respiratory complications are responsible for 50-67% of the morbidity in this population. OBJECTIVE: To investigate the effects of normocapnic hyperpnoea training in acute spinal cord injury. PATIENTS AND METHODS: Fourteen patients were randomized between control (sham) and an experimental normocapnic hyperpnoea training group. Vital capacity, maximal voluntary ventilation, respiratory muscle strength and endurance, respiratory complications and symptoms were evaluated before, after 4 and 8 weeks of training and after 8 weeks follow up. RESULTS: Maximal voluntary ventilation, respiratory muscle strength and endurance improved significantly in the experimental group compared with the control group (p < 0.05). Improvements in vital capacity tended to be different from the control group at 8 weeks of training. The Index of Pulmonary Dysfunction decreased after 4 weeks of training and respiratory complications were reported less frequently in the experimental group compared with the control group. CONCLUSION: Normocapnic hyperpnoea training in patients with spinal cord injury improved respiratory muscle strength and endurance. Respiratory complications occurred less frequently after training. PMID- 18509577 TI - Quality of life of traumatic spinal cord injured patients in Hong Kong. AB - OBJECTIVE: To assess the health-related quality of life of patients with traumatic spinal cord injury living in Hong Kong. DESIGN: A cross-sectional retrospective study. SUBJECTS: Subjects with paraplegia following traumatic spinal cord injury were recruited from our department, which is one of the 3 main regional spinal cord injury rehabilitation centres in Hong Kong. A total of 120 potential participants were invited to join the study. The final study group comprised 58 patients (25 (43%) men and 33 (57%) women, 30 (50%) of whom were under 40 years of age). METHODS: Health-related quality of life was assessed using the Hong Kong Chinese version of the World Health Organization Quality of Life Measure Abbreviated version (WHO-QOL-BREF(HK)). Results were summarized and compared with previous results from healthy and other disease populations in Hong Kong. RESULTS: There was no significant difference in quality of life between men and women, or between different durations of spinal cord injury, different educational levels or different marital status (p > 0.05). However, a significant difference in quality of life was found in different age groups and between people with incomplete and complete paraplegia (p < 0.05). Compared with healthy and other disease populations, people with paraplegia had the lowest quality of life in Hong Kong (p < 0.001). CONCLUSION: People with paraplegia in Hong Kong have a lower quality of life than the general Hong Kong population. More community support is required to meet the needs of these patients in the areas of transportation, medical care and social life, in particular. PMID- 18509578 TI - Manual dexterity in hereditary motor and sensory neuropathy type 1a: severity of limitations and feasibility and reliability of two assessment instruments. AB - OBJECTIVE: To assess the prevalence and significance of impaired manual dexterity in hereditary motor and sensory neuropathy type 1a (HMSN 1a), with the Sollerman hand function and the Functional Dexterity test, and compare the reliability and agreement of the tests. DESIGN: Descriptive cross-sectional study. SUBJECTS: Forty-nine subjects with HMSN 1a. RESULTS: Forty-six (94%) subjects had an abnormal Sollerman sum score (< 80) for the dominant hand. The most difficult subtests required finger grips such as pulp, tripod and lateral pinches. Dexterity scores of both hands were categorized as "moderately functional". Test retest reliability was excellent for the Sollerman test, with intraclass correlation coefficients between 0.98 and 0.99 (95% confidence interval (CI) 0.97 0.99), and good for Functional Dexterity test scores with correlation coefficients between 0.83 and 0.95 (95% CI. 71-0.97). The 95% limits of agreement between Sollerman tests showed that differences greater than 3 points can be interpreted as a change in dexterity. The Functional Dexterity test limits were wide. CONCLUSION: Impaired manual dexterity is common among subjects with HMSN 1a, stressing that the evaluation of dexterity is an essential element of the functional assessment. Both tests are able to detect impaired manual performance in HMSN 1a. For monitoring of disease progression and the effects of treatment programmes the Sollerman test is most suitable. PMID- 18509579 TI - A randomized controlled trial of aquatic and land-based exercise in patients with knee osteoarthritis. AB - OBJECTIVE: To compare the efficacy of aquatic exercise and a land-based exercise programme vs control in patients with knee osteoarthritis. METHODS: Primary outcome was change in pain, and in addition Knee Injury and Osteoarthritis Outcome Score questionnaire (KOOS). Standing balance and strength was also measured after and at 3-month follow-up. Seventy-nine patients (62 women), with a mean age of 68 years (age range 40-89 years) were randomized to aquatic exercise (n = 27), land-based exercise (n = 25) or control (n = 27). RESULTS: No effect was observed immediately after exercise cessation (8 weeks). At 3-month follow-up a reduction in pain was observed only in the land-based exercise group compared with control (-8.1 mm, (95% confidence interval -15.4 to -0.4; p = 0.039), but no differences between groups were observed for KOOS; and no improvement following aquatic exercise. Eleven patients reported adverse events (i.e. discomfort) in land-based exercise, while only 3 reported adverse events in the aquatic exercise. CONCLUSION: Only land-based exercise showed some improvement in pain and muscle strength compared with the control group, while no clinical benefits were detectable after aquatic exercise compared with the control group. However, aquatic exercise has significantly less adverse effects compared with a land based programme. PMID- 18509580 TI - Effects of deep and superficial heating in the management of frozen shoulder. AB - OBJECTIVES: To determine whether the addition of deep or superficial heating to stretching produces better clinical outcomes than stretching alone in the management of frozen shoulder. DESIGN: A single-blinded, randomized controlled study. SUBJECTS: Thirty subjects suffering from the stiffness phase of frozen shoulder. METHODS: Subjects were randomly allocated to receive: (i) deep heating plus stretching; (ii) superficial heating plus stretching; or (iii) stretching alone. Both heating groups received the respective treatments 3 times per week for 4 weeks. All groups received a standard set of shoulder stretching exercises. The American Shoulder and Elbow Surgeons assessment form was recorded at the baseline, sessions 6 and 12, and at the 4-week follow-up session. RESULTS: A significant improvement was seen in all groups in all outcome measures except for that of shoulder flexion range. The improvement in the shoulder score index and in the range of motion was significantly better in the deep heating group than in the superficial heating group. CONCLUSION: The addition of deep heating to stretching exercises produced a greater improvement in pain relief, and resulted in better performance in the activities of daily living and in range of motion than did superficial heating. PMID- 18509581 TI - Vitality, perceived social support and disease activity determine the performance of social roles in recently diagnosed multiple sclerosis: a longitudinal analysis. AB - OBJECTIVE: The aim of this study was to identify the principal determinants that are longitudinally associated with the performance of social roles in the first 3 years following a diagnosis of multiple sclerosis. DESIGN: Inception cohort with 5 measurements over 3 years. PATIENTS: A total of 156 patients recently diagnosed with multiple sclerosis. METHOD: Performance of social roles was measured using the 2 role functioning and the social sub-scales of the Medical Outcome Study Short Form 36. Potential determinants (n = 43) were divided into the following clusters: patient and disease characteristics (n = 12), psychosocial characteristics (n = 10), basic functions (n = 18) and basic activities (n = 3). Multivariate longitudinal regression analyses were performed with generalized estimating equations. A backwards selection procedure for every cluster per outcome reduced the large number of potential determinants. In order to determine whether longitudinal associations are present the selected determinants were entered into an overall regression model. RESULTS: Twenty-three candidate determinants were selected. Vitality, measured with the SF36 sub-scale vitality, the T2-weighted supratentorial lesion load and the perceived amount of social support, measured with the Social Support List Discrepancies, were longitudinally associated with the performance of social roles in 2 or 3 of the models. CONCLUSION: Vitality, the perceived amount of social support, and disease activity, i.e. the T2-weighted supratentorial lesion load, determine the performance of social roles in the early stages of multiple sclerosis. PMID- 18509582 TI - Copper toxicity on juveniles of Hyalella pseudoazteca Gonzalez and Watling, 2003. AB - The aim of this work is to evaluate the toxic effect of water and sediment contaminated with Cu(2+) on the survival and growth of juveniles of Hyalella pseudoazteca. The LC(50) 96 h for Cu(2+) in moderately hard water was 0.17 mg/L. The concentration of 100 mg Cu(2+)/kg in sediment was found to have an inhibitory growth effect. This study provides information on the toxic effects of Cu(2+) on a native benthic species occurring in the Pampean region, Argentina and contributes to validate, in this region, the interim use of Cu(2+) values recommended by the sediment quality guidelines for the Northern hemisphere. PMID- 18509583 TI - Discrepancies between physician interview and a patient self-assessment questionnaire after surgery for pelvic organ prolapse. AB - The objective of this article was to investigate whether there were discrepancies between the physician interview and a patient's self-assessment questionnaire on the functional results after surgery for pelvic organ prolapse. Before their follow-up appointment, 79 patients were sent a validated urogynaecological questionnaire. At the follow-up visit, the physician interviewed the patients using a checklist with ten symptoms and scored the items as present or absent. Seventy-two patients (91.1%) attended the follow-up visit and had completed the questionnaire. There was poor to slight agreement between the interview score and the self-reported responses to the questionnaire on all items. This was illustrated by low kappa coefficients. The physician was more optimistic about the outcome of the operation than was justified according to the answers to a patient self-assessment questionnaire. We therefore recommend the use of validated questionnaires to evaluate surgical outcome because they provide a more realistic (objective) view of the functional results. PMID- 18509584 TI - Necrotizing fasciitis following transobturator tape treated by extensive surgery and hyperbaric oxygen. AB - The transobturator sling procedure (TVT-O) was developed to minimize surgical risks involved in treating genuine stress incontinence. All data suggest that most risks associated with the retropubic route such as injuries to the bladder, intestines or vessels are practically obsolete with the obturator route. However, severe soft-tissue infections have been reported with this new technique. In this case report, necrotizing fasciitis (NF) developed shortly after a TVT-O procedure. This life-threatening complication required extensive debridements, a diverting colostomy, antibiotics, and eight sessions of hyperbaric oxygen (HBO) therapy. We emphasize the importance of a unified interdisciplinary clinical approach in severe NF with rapid progression and systemic toxemia. Primary, aggressive but tissue-saving debridements together with antibiotics are the cornerstones of therapy. HBO therapy can oxygenate infected hypoxic tissues to help marginally viable tissues survive, reduce the inflammatory response, improve leukocyte bacterial oxidative killing capacity, and achieve infection control and healing. PMID- 18509585 TI - Long-term patient satisfaction with prolapse surgery in general gynecology. AB - This retrospective cohort study aimed to explore the factors associated with long term success of prolapse surgery in a general gynecology setting. A chart review was performed of 528 women who had had a total of 535 operations for prolapse at least 4 years earlier. Those who had not had further pelvic floor surgery were sent a questionnaire exploring their satisfaction with surgery. The outcome could be determined for 406 (75%) of the women, of whom 238 (59%) were improved. In multivariate analyses, women were more likely to be satisfied with surgery if they were older, did not have a prior or concurrent colposuspension, if the prolapse operation included a hysterectomy and if they were operated on by one particular team. Selection of the right operation for the correct patient by the correct surgeon could improve the success of prolapse surgery. PMID- 18509586 TI - Cerebellar gangliocytoma presenting with hemifacial spasms: clinical report, literature review and possible mechanisms. AB - Cerebellar lesions have classically been considered not to cause epilepsy. However, previous reports have attributed seizures, beginning as hemifacial spasms to lesions of the cerebellar peduncles. We report an example of paroxysmal facial contractions associated with a cerebellar gangliocytoma. The seizures began on the first day of life and consisted of paroxysmal contractions involving the left orbicularis oculi, often the left forehead and lower facial muscles, sometimes accompanied by nystagmoid eye movements to the right and by head deviation to the left. Video-EEG monitoring showed only artifacts from muscle contractions. Magnetic resonance imaging showed a mass arising from the left superior cerebellar peduncle and partially occupying the fourth ventricle. The lesion was removed subtotally and partial seizure control was achieved. The neuropathological findings were consistent with a gangliocytoma. The literature in the association of cerebellar lesions with hemifacial spasms is reviewed and its possible mechanisms discussed. PMID- 18509587 TI - Anterior clinoidal meningiomas: analysis of 43 consecutive surgically treated cases. AB - BACKGROUND: Anterior clinoidal meningiomas are considered different from meningiomas of the medial sphenoid wing, but there is still some confusion about identification. The current classification scheme only considers tumour origin and invasion pattern around the clinoid process as indicators of resectability. However, the size of the tumour has important effects on surgical outcome. The purpose of this study was to analyze our clinical experience with anterior clinoidal meningiomas and refine the current classification scheme. METHOD: Forty three consecutive cases of anterior clinoidal meningioma were retrospectively analyzed. All were surgically treated at the Marmara University Department of Neurosurgery, the Marmara University Institute for Neurological Sciences, and the Department of Neurosurgery at Acibadem Hospital between 1987 and 2006. A pterional approach was used in all cases. FINDINGS: The mean tumour volume was 34.2 +/- 46.6 cc. Sixteen (37.2%) of the tumours were giant (largest diameter >4 cm). Total surgical removal was achieved in 39 cases (90.7%) and subtotal removal in 4 cases (9.3%). Eight patients (18.6%) developed early postoperative complications. The median and range of follow-up time was 39, R = 99 (3, 102) months. Four (9.3%) of the 43 tumours recurred during follow-up and were treated with radiosurgery. CONCLUSION: Anterior clinoidal meningioma is a separate disease entity from other meningiomas of the medial third of the sphenoid wing. Relatively good outcomes can be stated in this specific subgroup if growth criteria competable with true diagnosis are fulfilled. Size should be incorporated into any classification scheme for determining clinical and surgical risk. The standard pterional approach is sufficient for removing anterior clinoidal meningiomas. PMID- 18509588 TI - Intradural cement leakage: a rare complication of percutaneous vertebroplasty. AB - Percutaneous vertebroplasty (PV) is one of the alternative treatments for vertebral fractures. Reported significant complications include pain, radiculopathy, spinal cord compression, pulmonary embolism, infection and rib fractures. In this report, we highlight intradural cement leakage which is a rare complication of the procedure. A 49 year old man with a T12 compression fracture due to multiple myeloma was referred to the neurosurgery department from the orthopaedics and traumatology clinic after developing a right lower limb weakness following percutaneous vertebroplasty with polymethylmethacrylate. An urgent thoraco-lumbar magnetic resonance imaging was performed. The T1 and T2-weighted images demonstrated intradural extramedullary and epidural cement leakages which were hypointense on both sequences. Total laminectomy was performed at T12 and L1 and two epidural cement collections were excised on the right. Then, a dural incision from T12 to the body of L1 was done and cement material seen in front of the rootlets excised without any nerve injury. The patient was discharged after a week and referred to the haematology clinic for additional therapy of multiple myeloma. Although the cement leakage was extensive, the right leg weakness improved significantly and he began to walk with assistance 3 months later. Good quality image monitoring and clear visualisation of cement are essential requirements for PV using polymethylmethacrylate to prevent this complication from the treatment. PMID- 18509589 TI - Endovascular coil transfixing a cranial nerve five years after embolisation. AB - After endovascular coiling, several authors have reported refilling of the aneurysm, appearance of a mass effect, coils protruding into the parent artery, migration of coils into parent artery or through aneurysm wall, and compression of the parent artery by coil impaction. This is the first report of an endovascular coil transfixing a cranial nerve. We present a 59 year old man who presented with a symptomatic bilobulated aneurysm of the right internal carotid artery. The aneurysm was embolised by endovascular coiling. Angiographic follow up showed occlusion of the aneurysm. Five years later, the patient complained of progressive diplopia with ptosis. Follow-up angiography showed renewed filling of the aneurysm at its neck. The aneurysm was clipped surgically. At operation, a coil mass effect was noted and one coil penetrated the fibres of the right oculomotor nerve. PMID- 18509590 TI - Molecular characterization of Lolium latent virus, proposed type member of a new genus in the family Flexiviridae. AB - Lolium latent virus (LoLV) was recently detected in the USA for the first time in ryegrass hybrids (Lolium perenne x Lolium multiflorum). The genome of one USA isolate, LoLV-US1, has now been fully sequenced. The positive strand genomic RNA is 7674 nucleotides (nt) long and is organized in five open reading frames (ORFs) encoding the replication-associated protein, the movement-associated triple gene block proteins and the coat protein (CP). The genome organization is similar to that of viruses in the genera Potexvirus and Foveavirus; however, analysis of the complete LoLV genomic sequence, phylogenetic analyses of the deduced amino acid (aa) sequences of the polymerase and the CP, presence of a putative ORF 6, and the in vivo detection of two CPs in equimolar amounts, highlight features peculiar to LoLV. These characteristics indicate that LoLV forms a monotypic group separate from existing genera and unassigned species within the family Flexiviridae, for which we propose the genus name Lolavirus. One-step RT-PCR was developed for quick and reliable LoLV detection. PMID- 18509591 TI - Effect of taurine on alcoholic liver disease in rats. AB - To investigate the effect of taurine on alcoholic liver disease in rats, male Wistar rats were administered alcohol intragastrically for 3 months. The effect of beta-alanine-mediated taurine depletion and taurine administration on the development of alcoholic liver disease was examined. It was found that taurine administration produced lower levels of aspartate aminotransferase and alkaline aminotransferase than that of the untreated group. In addition, the levels of hepatic total protein, glutathione and superoxide dismutase were higher in the taurine treated groups than those in the untreated control or the taurine depleted groups, while hepatic malondialdehyde content exhibited the negative effect. Moreover, the concentrations of hepatic hydroxyproline, serum hyaluronic acid, interleukin-2, interleukin-6, tumor necrosis factor-alpha and laminin were all decreased in the taurine treated groups. The pathological changes showed that the percentage of fatty degeneration and inflammation in the taurine groups were lower than that of the control, taurine depleted and automatic recovery groups. These in vivo findings demonstrate that hepatic disease caused by chronic alcohol consumption can be prevented and cured by administration of taurine. PMID- 18509592 TI - Postoperative results and complications of total elbow arthroplasty in patients with rheumatoid arthritis: three types of nonconstrained arthroplasty. AB - Postoperative results and complications of total elbow arthroplasty (TEA) conducted for rheumatoid arthritis (RA) patients at our institute were studied. Primary TEAs were performed in 72 patients. The mean follow-up period was 3.5 years. Three types of prostheses were implanted: JACE prosthesis in 34 elbows, STABLE prosthesis in 13 elbows, and KUDO prosthesis (type 5) in 32 elbows. The outcome was evaluated by the change in the range of motion and the Japanese Orthopaedic Association functional evaluation score for the elbow joint (JOA score). The arc of motion and the JOA score at discharge and at final examination significantly improved in patients with the three types of prosthesis. The loosening rates for the JACE, STABLE and KUDO prostheses were 15, 23, and 0%, respectively, although the follow-up periods were different. The loosening rate decreased to 2.5% when the humeral component was fixed with cement. Intraoperative fractures occurred in eight (10.1%) elbows and ulnar nerve palsy in six. Deep infection developed in three (4.8%) elbows and was treated by removing the prosthesis. Although there were considerable complications, the marked improvements in pain and function favor TEA in patients with rheumatoid elbow. PMID- 18509593 TI - Rich polymorphism of a rod-like liquid crystal (8CB) confined in two types of unidirectional nanopores. AB - We present a neutron and X-rays scattering study of the phase transitions of 4-n octyl-4' -cyanobiphenyl (8CB) confined in unidirectional nanopores of porous alumina and porous silicon (PSi) membranes with an average diameter of 30 nm. Spatial confinement reveals a rich polymorphism, with at least four different low temperature phases in addition to the smectic A phase. The structural study as a function of thermal treatments and conditions of spatial confinement allows us to get insights into the formation of these phases and their relative stability. It gives the first description of the complete phase behavior of 8CB confined in PSi and provides a direct comparison with results obtained in bulk conditions and in similar geometric conditions of confinement but with reduced quenched disorder effects using alumina anopore membranes. PMID- 18509594 TI - Responses of Rhodotorula sp. Y11 to cadmium. AB - Some aspects of the cellular responses to cadmium were extensively investigated in the yeast Rhodotorula sp. Y11. Scanning electron microscopy indicated that accumulation of cadmium in the Y11 did not cause any visible effects on cell morphology. More than 20% yeast cells still showed viability after 15 h of cadmium accumulation under 100 mg l(-1) cadmium concentration, and transmission electron microscopy analysis showed that plasmolysis and thickened cell wall were not observed in all of the cells. In the presence of cadmium, the activities of superoxide dismutase (SOD) and catalase (CAT) were all greater than the control, but the increase was in a dose-independent manner. Changes in SOD and CAT activities were also dependent on the time of exposure. Therefore, it suggests that antioxidative defenses play an important role in cadmium tolerance in Rhodotorula sp. Y11. Nondenaturing polyacrylamide gels revealed only one SOD isoforms in Y11 even under exposure to cadmium. PMID- 18509595 TI - Proteomic and molecular investigation on the physiological adaptation of Comamonas sp. strain CNB-1 growing on 4-chloronitrobenzene. AB - Comamonas sp. strain CNB-1 can utilize 4-chloronitrobenzene (4CNB) as sole carbon and nitrogen source for growth. Previous studies were focused on 4CNB degradative pathway and have showed that CNB-1 contained a plasmid pCNB1 harboring the genes (cnbABCaCbDEFGH, cnbZ) for the enzymes involving in 4CNB degradation, but only three gene products (CnbCa, CnbCb, and CnbZ) were identified in CNB-1 cells. Comamonas strain CNB-2 that lost pCNB1 was not able to grow on 4CNB. In this study, physiological adaptation to 4CNB by CNB-1 was investigated with proteomic and molecular tools. Comparative proteomes of strains CNB-1 and CNB-2 grown on 4CNB and/or succinate revealed that adaptation to 4CNB by CNB-1 included specific degradative pathway and general physiological responses: (1) Seven gene products (CnbA, CnbCa, CnbCb, CnbD, CnbE, CnbF, and CnbZ) for 4CNB degradation were identified in 4CNB-grown cells, and they were constitutively synthesized in CNB 1. Two genes cnbE and cnbF were cloned and simultaneously expressed in E. coli. The CnbE and CnbF together catalyzed the conversion of 2-oxohex-4-ene-5-chloro 1,6-dioate into 2-oxo-4-hydroxy-5-chloro-valeric acid; (2) Enzymes involving in glycolysis, tricarboxylic acid cycle, and synthesis of glutamate increased their abundances in 4CNB-grown cells. PMID- 18509596 TI - Elastase-sensitive elastomeric scaffolds with variable anisotropy for soft tissue engineering. AB - PURPOSE: To develop elastase-sensitive polyurethane scaffolds that would be applicable to the engineering of mechanically active soft tissues. METHODS: A polyurethane containing an elastase-sensitive peptide sequence was processed into scaffolds by thermally induced phase separation. Processing conditions were manipulated to alter scaffold properties and anisotropy. The scaffold's mechanical properties, degradation, and cytocompatibility using muscle-derived stem cells were characterized. Scaffold in vivo degradation was evaluated by subcutaneous implantation. RESULTS: When heat transfer was multidirectional, scaffolds had randomly oriented pores. Imposition of a heat transfer gradient resulted in oriented pores. Both scaffolds were flexible and relatively strong with mechanical properties dependent upon fabrication conditions such as solvent type, polymer concentration and quenching temperature. Oriented scaffolds exhibited anisotropic mechanical properties with greater tensile strength in the orientation direction. These scaffolds also supported muscle-derived stem cell growth more effectively than random scaffolds. The scaffolds expressed over 40% weight loss after 56 days in elastase containing buffer. Elastase-sensitive scaffolds were complete degraded after 8 weeks subcutaneous implantation in rats, markedly faster than similar polyurethanes that did not contain the peptide sequence. CONCLUSION: The elastase-sensitive polyurethane scaffolds showed promise for application in soft tissue engineering where controlling scaffold mechanical properties and pore architecture are desirable. PMID- 18509597 TI - Conversion of nanosuspensions into dry powders by spray drying: a case study. AB - PURPOSE: Drying of nanosuspensions can cause destabilization of the particles, leading to irreversible aggregation. In order to prepare an effective solid dosage form for a nanosuspension, it is imperative that the spray-dried nanoparticles should go back to their original particle size when reconstituted in an aqueous system. This case study examines impact of various formulation and processing parameters on redispersibility of the spray dried nanoparticles. METHODS: Nanosuspensions were prepared using the microprecipitation homogenization process. Spray drying of nanosuspensions was achieved using a lab scale Buchi spray dryer. RESULTS: Formulation components appeared to have the most significant impact on redispersibility of spray dried particles. Absence of surface charge led to particles that could not be redispersed. On the other hand, charged particles stabilized with an appropriate sugar led to spray dried powders that were flowable and easily redispersible. Dissolution testing showed the presence of two phases--a lag phase that represented dispersion of the loose aggregates, and dissolution of the dispersed nanoparticles. CONCLUSIONS: Nanosuspensions of a poorly soluble drug could be spray dried to obtain flowable powders that could be easily redispersed. These optimized powders also showed significantly improved dissolution rates as compared to the micronized drug, or unoptimized nanosuspensions. PMID- 18509598 TI - Investigations into the formulation of metered dose inhalers of salmeterol xinafoate and fluticasone propionate microcrystals. AB - PURPOSE: To investigate the aerosolization and behaviour of microparticles of salmeterol xinafoate (SX) and fluticasone propionate (FP) suspended in hydrofluoroalkane (HFA) propellant. METHODS: Microcrystals of SX and FP were produced from poly(ethylene glycol) by antisolvent crystallization. The suspension behaviour and aerosolization of the microcrystals when formulated as metered dose inhalers (MDIs) in HFA 134a propellant was compared with that of microparticles produced by micronization (mSX and mFP) using a glass twin stage impinger and by laser light diffraction using a pressurized cell. RESULTS: FP microparticles underwent non-reversible aggregation in suspension as seen by a doubling in the volume median diameter compared to the raw material. The degree of aggregation of SX particles in suspension was found to decrease as the particle size of the original particles increased. However, because the SX aggregate size was lowest for the particles with the smallest initial size (mSX), the highest fine particle fraction (FPF) of SX was obtained from a suspension of mSX. The FPFs following aerosolization of FP suspensions were similar although the FPF was lowest for particles with the largest original size. CONCLUSIONS: The size of the aggregates in the HFA suspensions was found to correlate directly with the FPFs determined by impaction. PMID- 18509599 TI - Diammine dicarboxylic acid platinum enhances cytotoxicity in platinum-resistant ovarian cancer cells through induction of apoptosis and S-phase cell arrest. AB - PURPOSE: Polysaccharides such as chondroitin play a potent role in tumor growth, tissue repair and angiogenesis. These properties make chondroitin a good candidate for novel drug delivery systems. Diammine dicarboxylic acid platinum (DDAP), a novel polymeric platinum compound, was developed by conjugating the platinum analogue to aspartate-chondroitin for drug delivery to tumor cells. DDAP improves platinum solubility which may reduce systemic toxicity and be more efficacious than cisplatin in killing tumor cells. METHODS: We tested and compared the cytotoxic effects of DDAP and CDDP on the platinum-sensitive 2008 and A2780 ovarian cancer cell lines and their platinum-resistant sublines 2008.C13 and A2780cis; we also investigated DDAP's mechanism of action. RESULTS: In the platinum-sensitive cell lines, the cytotoxic effects of DDAP and CDDP were comparable. However, in the platinum-resistant sublines, significantly greater cell-growth inhibition was induced by DDAP than by CDDP, especially at lower doses. DDAP also induced more apoptosis than CDDP did in the 2008.C13 subline, which was partially mediated by the caspase 3-dependent pathway. In addition, lower (but not higher) doses of DDAP arrested 90% of S-phase 2008.C13 cells, which might be associated with up-regulation of p21 and maintenance of low cyclin A expression. Furthermore, greater cellular uptake of DDAP was seen in platinum resistant than in platinum-sensitive ovarian cancer cells. CONCLUSIONS: Low-dose DDAP enhances drug delivery to platinum-resistant ovarian cancer cells and substantially inhibits their growth by inducting apoptosis and arresting cells in the S-phase, suggesting that DDAP may overcome platinum resistance in ovarian cancer. PMID- 18509600 TI - Designing 3D photopolymer hydrogels to regulate biomechanical cues and tissue growth for cartilage tissue engineering. AB - PURPOSE: Synthetic hydrogels fabricated from photopolymerization are attractive for tissue engineering for their controlled macroscopic properties, the ability to incorporate biological functionalities, and cell encapsulation. The goal of the present study was to exploit the attractive features of synthetic hydrogels to elucidate the role of gel structure and chemistry in regulating biomechanical cues. METHODS: Cartilage cells were encapsulated in poly(ethylene glycol) (PEG) hydrogels with different crosslinking densities. Cellular deformation was examined as a function of gel crosslinking. The effects of continuous versus intermittent dynamic loading regimens were examined. RGD, a cell adhesion peptide, was incorporated into PEG gels and subjected to mechanical loading. Chondrocyte morphology and activity was assessed by anabolic and catabolic ECM gene expression and matrix production by collagen and glycosaminoglycan production. RESULTS: Cell deformation was mediated by gel crosslinking. In the absence of loading, anabolic activity was moderately upregulated while catabolic activity was significantly inhibited regardless of gel crosslinking. Dynamic loading enhanced anabolic activities, but continuous loading inhibited catabolic activity, while intermittent loading stimulated catabolic activity. RGD acted as a mechanoreceptor to influence tissue deposition. CONCLUSIONS: We demonstrate the ability to regulate biomechanical cues through manipulations in the gel structure and chemistry and cartilage tissue engineering. PMID- 18509601 TI - Organic-inorganic surface modifications for titanium implant surfaces. AB - This paper reviews current physicochemical and biochemical coating techniques that are investigated to enhance bone regeneration at the interface of titanium implant materials. By applying coatings onto titanium surfaces that mimic the organic and inorganic components of living bone tissue, a physiological transition between the non-physiological titanium surface and surrounding bone tissue can be established. In this way, the coated titanium implants stimulate bone formation from the implant surface, thereby enhancing early and strong fixation of bone-substituting implants. As such, a continuous transition from bone tissue to implant surface is induced. This review presents an overview of various techniques that can be used to this end, and that are inspired by either inorganic (calcium phosphate) or organic (extracellular matrix components, growth factors, enzymes, etc.) components of natural bone tissue. The combination, however, of both organic and inorganic constituents is expected to result into truly bone-resembling coatings, and as such to a new generation of surface modified titanium implants with improved functionality and biological efficacy. PMID- 18509602 TI - Click chemistry, a powerful tool for pharmaceutical sciences. AB - Click chemistry refers to a group of reactions that are fast, simple to use, easy to purify, versatile, regiospecific, and give high product yields. While there are a number of reactions that fulfill the criteria, the Huisgen 1,3-dipolar cycloaddition of azides and terminal alkynes has emerged as the frontrunner. It has found applications in a wide variety of research areas, including materials sciences, polymer chemistry, and pharmaceutical sciences. In this manuscript, important aspects of the Huisgen cycloaddition will be reviewed, along with some of its many pharmaceutical applications. Bioconjugation, nanoparticle surface modification, and pharmaceutical-related polymer chemistry will all be covered. Limitations of the reaction will also be discussed. PMID- 18509603 TI - In vitro and in vivo investigation on PLA-TPGS nanoparticles for controlled and sustained small molecule chemotherapy. AB - PURPOSE: The aim of this work was to evaluate in vivo poly(lactide)-D: -alpha tocopheryl polyethylene glycol 1,000 succinate nanoparticles (PLA-TPGS NPs) for controlled and sustained small molecule drug chemotherapy. METHODS: The drug loaded PLA-TPGS NPs were prepared by the dialysis method. Particle size, surface morphology and surface chemistry, in vitro drug release and cellular uptake of NPs were characterized. In vitro and in vivo therapeutic effects of the nanoparticle formulation were evaluated in comparison with Taxol. RESULTS: The PLA-TPGS NP formulation exhibited significant advantages in in vivo pharmacokinetics and xenograft tumor model versus the PLGA NP formulation and the pristine drug. Compared with Taxol, the PLA-TPGS NP formulation achieved 27.4 fold longer half-life in circulation, 1.6-fold larger area-under-the-curve (AUC) with no portion located above the maximum tolerance concentration level. For the first time in the literature, one shot for 240 h chemotherapy was achieved in comparison with only 22 h chemotherapy for Taxol at the same 10 mg/kg paclitaxel dose. Xenograft tumor model further confirmed the advantages of the NP formulation versus Taxol. CONCLUSIONS: The PLA-TPGS NP formulation can realize a way of controlled and sustained drug release for more than 10 days, which relieves one of the two major concerns on cancer nanotechnology, i.e. feasibility. PMID- 18509604 TI - Use of tc-99m mebrofenin as a clinical probe to assess altered hepatobiliary transport: integration of in vitro, pharmacokinetic modeling, and simulation studies. AB - PURPOSE: Transport of the hepatobiliary scintigraphy agent Tc-99m mebrofenin (MEB) was characterized and simulation studies were conducted to examine the effects of altered hepatic transport on MEB pharmacokinetics in humans. METHODS: MEB transport was investigated in Xenopus laevis oocytes expressing OATP1B1 or OATP1B3, and in membrane vesicles prepared from HEK293 cells transfected with MRP2 or MRP3. A pharmacokinetic model was developed based on blood, urine and bile concentration-time profiles obtained in healthy humans, and the effect of changes in hepatic uptake and/or excretion associated with disease states (hyperbilirubinemia and cholestasis) on MEB disposition was simulated. RESULTS: MEB (80 pM) transport by OATP1B1 and OATP1B3 was inhibited by rifampicin (50 microM) to 10% and 4% of control, respectively. MEB (0.4 nM) transport by MRP2 was inhibited to 12% of control by MK571 (50 microM); MRP3-mediated transport was inhibited to 5% of control by estradiol-17-beta-glucuronide (100 microM). A two compartment model described MEB (2.5 mCi) systemic disposition in humans (systemic clearance = 16.2 +/- 2.7 ml min(-1) kg(-1)); biliary excretion was the predominant route of hepatic elimination (efflux rate constants ratio canalicular/sinusoidal = 3.4 +/- 0.8). Based on simulations, altered hepatic transport markedly influenced MEB systemic and hepatic exposure. CONCLUSIONS: MEB may be a useful probe to assess how altered hepatic function at the transport level modulates hepatobiliary drug disposition. PMID- 18509605 TI - Hyoscyamine biosynthesis in Datura stramonium hairy root in vitro systems with different ploidy levels. AB - Hyoscyamine biosynthesis in Datura stramonium hairy roots with different ploidy levels was investigated. For the first time, we report that hairy roots undergo endoreduplication and therefore consist mainly of cells with doupled sets of chromosomes of primary plant tissues, used for Agrobacterium transformation. The alkaloid profiles of hairy roots obtained from diploid and tetraploid plants were similar in terms of the major compounds, but they differed significantly with respect to the minor compounds (here defined as those that accounted for <1% of the total ion current of the alkaloid mixture in gas chromatography-mass spectrometric analyses). Significant differences in the effects of the main nutrients on the growth of the hairy roots obtained from diploid and tetraploid plants and their hyoscyamine contents were observed. The maximal yield of hyoscyamine (177 mg/L) was obtained when hairy roots from tetraploid plants were cultivated in Murashige-Skoog nutrient medium supplemented with 6% sucrose. Time courses of utilization of the main nutrients in the medium during cultivation of D. stramonium hairy root cultures are also presented. PMID- 18509606 TI - Superoxide dismutase-loaded PLGA nanoparticles protect cultured human neurons under oxidative stress. AB - The objective of our study was to investigate the neuroprotective efficacy of superoxide dismutase (SOD), loaded in poly(D,L-lactide co-glycolide; PLGA) nanoparticles (NPs), in cultured human neurons challenged with hydrogen peroxide (H(2)O(2))-induced oxidative stress. We hypothesized that the protected and sustained intracellular delivery of SOD encapsulated in NPs would demonstrate better neuroprotection from oxidative stress than either SOD or pegylated SOD (PEG-SOD) in solution. SOD-NPs (approximately 81 +/- 4 nm in diameter, 0.9% w/w SOD loading) released the encapsulated SOD in an active form with 8.2% cumulative release during the first 24 h, followed by a slower release thereafter. The results demonstrated that PLGA-NPs are compatible with human neurons, and the neuroprotective effect of SOD-NPs is dose-dependent, with efficacy seen at >100 U SOD, and less significant effects at lower doses. Neither SOD (25-200 U) nor PEG SOD (100 U) in solution demonstrated the neuroprotective effect under similar conditions. The neuroprotective effect of SOD-NPs was seen up to 6 h after H(2)O(2)-induced oxidative stress, but the effect diminished thereafter. Confocal microscopic studies demonstrated better intracellular neuronal uptake of the encapsulated model protein (fluorescein isothiocyanate-labeled BSA) than the protein in solution. Thus, the mechanism of efficacy of SOD-NPs appears to be due to the stability of the encapsulated enzyme and its better neuronal uptake after encapsulation. PMID- 18509607 TI - Vermicomposting of solid waste generated from leather industries using epigeic earthworm Eisenia foetida. AB - Animal fleshing (ANFL) generated as solid waste from tannery industries was vermicomposted using the epigeic earthworm Eisenia foetida. The mixing ratio of ANFL with cow dung and agricultural residues as feed mixtures was maintained to be 3:1:1 respectively during the vermicomposting experiments for 50 days. Vermicomposting resulted in the reduction of pH 6.74 and C:N ratio 15.5 compared to the control sample. A notable increase in earthworm biomass was also observed in the vermin bioreactor. The germination index of 84% for tomato seedlings (Lycopersicon esculentum cv. PKM1) was observed for the vermicomposted soil. Scanning electron microscope and Fourier transform infrared spectroscopy were recorded to identify the changes in surface morphology and functional groups in the control and vermicomposted samples. The results obtained from the present study indicated that the earthworm E. foetida was able to convert ANFL into nutrient-enriched products. PMID- 18509608 TI - Cloning of Qiantang River triangular bream (Megalobrama terminalis) IGF-I gene and expression of the recombinant pre-IGF-I in Escherichia coli. AB - The insulin-like growth factor (IGF-I) gene (GenBank accession no. AY247412) of Qiantang River triangular bream (Megalobrama terminalis) was cloned for the first time from the liver by reverse transcriptase polymerase chain reaction. The gene was inserted into pMD 18-T vector to construct the recombinant plasmid pMD 18 T/IGF-I. Sequence analysis indicated that the IGF-I cDNA consisted of 486 nucleotides encoding 161 amino acids, which spanned the complete signal peptide, mature peptide (including B, C, A, and D domains), and E-domain. Analysis of the E domain indicated that triangular bream IGF-I gene belonged to the IGF-I Ea-2 subtype. To construct the expression plasmid, the IGF-I gene was subcloned into prokaryotic expressing vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/IGF-I was transformed into Escherichia coli BL21 (DE3), and the transgene expression was observed after being induced with isopropylthiogalactoside. The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity, and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin-layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3, and 18.8% of total bacterial protein, respectively. PMID- 18509609 TI - [Video-assisted patient education in anaesthesiology: possibilities and limits of a new procedure for improvement of patient information]. AB - In video-assisted patient education (ViPa), patients watch an educational video about the process and the risks of anaesthesia in addition to the preanaesthetic interview with the anaesthesiologist. Used as a supplement to the preanaesthetic visit, the videos can increase patients' knowledge and satisfaction without having any negative effect on perioperative anxiety. Because the video graphically depicts the basic information, the preanaesthetic visit can then focus on specific aspects of the individual patient, i.e. high anxiety or specific questions. The redundant and monotonous explanations about the procedures and risks of anaesthesia by the interviewing anaesthesiologist are partly replaced by the video, but for medico-legal reasons the ViPa cannot totally replace the preanesthetic interview. It can be used in pediatric anaesthesia and reduces parental anxiety. Because of the lack of studies, the effects of the ViPa on perioperative patient compliance, especially for outpatient surgery, and on the economics of anaesthesia clinics are unclear. PMID- 18509610 TI - [Surgical voice rehabilitation in unilateral vocal fold paralysis]. AB - Unilateral recurrent nerve paralysis leads to glottic insufficiency, significantly reducing vocal ability. Due to its unusually long course, the recurrent laryngeal nerve is prone to iatrogenic lesions involves many medical fields generally with little expertise in voice disorders. Whenever the etiology is uncertain, a complete diagnostic work-up is mandatory. Indirect laryngoscopy confirms the diagnosis. Laryngeal electromyography is of great value because it differentiates between paralysis and ankylosis of the cricoarytenoid joint. Moreover in many cases laryngeal electromyography yields a reliable prognosis of clinical outcome. While unfavorable results can be predicted with high accuracy, correct prognosis of complete recovery is more difficult. Speech therapy is the treatment of choice in cases of unilateral recurrent nerve palsy. Patients with persistent glottal gap may express the wish for surgical voice rehabilitation. Nowadays a broad spectrum of endoscopic and open approaches are available for this purpose. This review describes advanced techniques of voice-improving surgery available in specialized centers today. PMID- 18509611 TI - A standardized perioperative surgical site infection care process among children with stoma closure: a before-after study. AB - BACKGROUND: We report on the effectiveness of a standardized perioperative care process for lowering surgical site infection (SSI) rates among children with stoma closure at a tertiary-care public pediatric teaching hospital in Mexico City. METHODS: All consecutive children with stoma closure operated on between November 2003 and October 2005 were prospectively followed for 30 days postoperatively. We conducted a before-after study to evaluate standardized perioperative bowel- and abdominal-wall care process results on SSI rates. RESULTS: Seventy-one patients were operated on, and all completed follow-up. SSI rates declined from 42.8% (12/28) before to 13.9% (6/43) after the standardization procedure (relative risk (RR) = 3.1; 95% confidence interval (CI) = 1.3-7.2; p = 0.006). SSI independently associated risk factors comprised peristomal skin inflammation >3 mm (odds ratio (OR) = 9.6; 95% CI = 1.8-49.6; p = 0.007) and intraoperative complications (OR = 13.3; 95% CI = 1.4-127.2; p = 0.02). Being operated on during the after-study period was shown to be a protective factor against SSI (OR = 0.2; 95% CI = 0.4-0.97; p = 0.04). CONCLUSION: Standardization was able to reduce SSI rates threefold in children with stoma closure in a short period of time. PMID- 18509612 TI - [Cardiac arrhythmias in the pregnant woman and the fetus]. AB - For the acute treatment of supraventricular (SVT) and ventricular tachycardias (VT) in pregnant women, electrical cardioversion with 50-100 J is indicated in all unstable patients. In stable SVT the initial therapy includes vagal maneuvers or intravenous adenosine. For long-term therapy, beta-blocking agents with beta(1)-selectivity are first-line drugs or specific anti-arrhythmic drugs. An implantable cardioverter-defibrillator is another approach. In patients with symptomatic bradycardia, a pacemaker can be implanted using echocardiography at any stage of pregnancy. Evaluation of fetal arrhythmias in clinical practice is based on assessment of the chronological relationship between atrial and ventricular contraction (M-mode and Doppler ultrasound or magnetocardiography). Most forms of SVT can be treated with transplacental administration of anti arrhythmic drugs. Atrioventricular (AV) block in fetuses with structural heart disease is frequently associated with hydrops fetalis and intrauterine death. Administration of corticoids and beta-mimetic drugs is used to treat antibody mediated AV block and cardiomyopathy. PMID- 18509613 TI - [Hypertensive diseases in pregnancy]. AB - Hypertension affects about 10% of pregnancies and is a leading cause of both maternal and fetal morbidity and mortality worldwide. Hypertension in pregnancy includes a spectrum of conditions, including preeclampsia or eclampsia, chronic arterial hypertension, gestational hypertension, and pulmonary hypertension. In this review article, current data and recommendations are summarized regarding definitions, diagnoses, and treatment options for hypertensive disorders in pregnancy. PMID- 18509614 TI - [What does the obstetrician expect from the internist during prenatal care?]. AB - Cooperation between the obstetrician and the physician for internal medicine is especially needed when a pregnant woman suffers from complex medical diseases. This contribution focuses on the special circumstances of maternal treatment also considering the fetal state with the following examples: drugs in the first trimester, cardiac diseases, pregnancy-induced hypertension, HELLP syndrome, gestational and type I diabetes, thrombosis, pulmonary diseases, infections, and treatment of cancer during pregnancy. Special obstetric requests to the physician are: keeping in close contact, avoiding recommendations for the mode of delivery, avoiding overtreatment of hypertensive disorders as well as intensifying the surveillance including the creation of preventive schedules for pregnant women with an increased risk of developing metabolic syndrome in their later life. PMID- 18509615 TI - Shocking effects of endothelial bradykinin B1 receptors. PMID- 18509616 TI - A new azhdarchoid pterosaur from the Lower Cretaceous of China and its implications for pterosaur phylogeny and evolution. AB - Toothless pterosaurs played a key role in broadening the taxonomic, morphological and ecological diversity of Cretaceous pterosaurs. Here we report a complete, articulated skeleton of a 1.4-m-wingspan pterosaur from the Lower Cretaceous Jiufotang Formation of Liaoning Province, China which is identified as a new genus and species, Shenzhoupterus chaoyangensis gen. et sp. nov. The new taxon is edentulous, with a relatively large skull and a remarkably large, tall nasoantorbital fenestra that extends well above the main part of the braincase. Phylogenetic analysis shows that Shenzhoupterus gen. nov. belongs in a distinct clade of azhdarchoid pterosaurs, formally recognised here as a new family, Chaoyangopteridae, that also includes Chaoyangopterus, Jidapterus and Eoazhdarcho from the Jiufotang Formation and Eopteranodon from the Yixian Formation. These new data clarify recent confusion surrounding the systematics of these Lower Cretaceous taxa and provide new insights into the evolutionary history of pterosaurs. PMID- 18509618 TI - [Combined hip-dysplasia and femuroacetabular impingement. Diagnosis and simultaneous surgical treatment]. AB - Developmental dysplasia of the hip as well as femuroacetabular impingement are recognized causes of hip pain and degenerative disorders. While accepted treatment recommendations for both entities currently exist, there are no standardised approaches published for the management of combined lesions. We therefore present an algorithm of diagnosis and simultaneous surgical treatment based on a combination of periacetabular osteotomy and arthroscopically-assisted minimally invasive anterior osteochondroplasty. Short-term treatment results of 19 patients show an effective improvement of clinical symtoms and radiographic deformity. PMID- 18509619 TI - Non-invasive assessment of intracranial pressure using ocular sonography in neurocritical care patients. AB - OBJECTIVE: To assess the relationship between optic nerve sheath diameter (ONSD) and intracranial pressure (ICP) in neurocritical care patients. DESIGN: Prospective, observational study. SETTING: Surgical critical care unit, level 1 trauma center. PATIENTS: A total number of 37 adult patients requiring sedation and ICP monitoring after severe traumatic brain injury, subarachnoid hemorrhage, intracranial hematoma, or stroke. MEASUREMENTS AND MAIN RESULTS: Optic nerve sheath diameter was measured with a 7.5 MHz linear ultrasound probe. ICP was measured invasively via a parenchymal device. Simultaneous measurements were performed at least once a day during the first 2 days after ICP insertion and in cases of acute changes. There was a significant relationship between ONSD and ICP (78 simultaneous measures, r = 0.71, P < 0.0001). Changes in ICP were strongly correlated with changes in ONSD (39 measures, r = 0.73, P < 0.0001). Enlarged ONSD was a suitable predictor of elevated ICP (>20 mmHg) (area under ROC curve = 0.91). When ONSD was less than 5.86 mm, the negative likehood ratio for raised ICP was 0.06. CONCLUSION: In sedated neurocritical care patients, non-invasive sonographic measurements of ONSD are correlated with invasive ICP, and the probability to have raised ICP if ONSD is less than 5.86 mm is very low. This method could be used as a screening test when raised ICP is suspected. PMID- 18509620 TI - Benfotiamine reduces genomic damage in peripheral lymphocytes of hemodialysis patients. AB - Hemodialysis patients have an elevated genomic damage in peripheral blood lymphocytes (PBLs) and an increased cancer incidence, possibly due to accumulation of uremic toxins like advanced glycation end products (AGEs). Because the vitamin B1 prodrug benfotiamine reduces AGE levels in experimental diabetes, and dialysis patients often suffer from vitamin B1 deficiency, we conducted two consecutive studies supplementing hemodialysis patients with benfotiamine. In both studies, genomic damage was measured as micronucleus frequency of PBLs before and at three time-points after initiation of benfotiamine supplementation. AGE-associated fluorescence in plasma, and in the second study additionally, the antioxidative capacity of plasma was analyzed. Benfotiamine significantly lowered the genomic damage of PBLs in hemodialysis patients of both studies independent of changes in plasma AGE levels. The second study gave a hint to the mechanism, as the antioxidative capacity of the plasma of the treated patients clearly increased, which might ameliorate the DNA damage. PMID- 18509621 TI - Attenuation of cocaine self-administration in squirrel monkeys following repeated administration of the mGluR5 antagonist MPEP: comparison with dizocilpine. AB - RATIONALE: The mGluR5 antagonist MPEP has effects that suggest potential as a pharmacotherapy for cocaine addiction. MPEP can attenuate self-administration of cocaine in animals; however, studies usually involved only acute treatment with MPEP and a single dose of self-administered cocaine. Cocaine addicts use varied amounts of cocaine over long periods of time, and an effective pharmacotherapy would almost certainly require more chronic treatment. OBJECTIVES: The present study (1) compared the effects of repeated treatment with MPEP or the NMDA receptor antagonist dizocilpine on the reinforcing effects of a range of doses of cocaine and (2) determined the pharmacological specificity of the effects of the drugs in attenuating cocaine self-administration compared to food-reinforced behavior. An effective pharmacotherapy should selectively reduce cocaine self administration. MATERIALS AND METHODS: Groups of monkeys responded under a fixed ratio schedule of i.v. cocaine self-administration or food-pellet delivery. The effects of daily treatment with MPEP and dizocilpine were determined under both the schedule of i.v. cocaine injection and food delivery. RESULTS: Treatment with MPEP and dizocilpine significantly reduced cocaine self-administration, producing rightward and downward shifts in the ascending limb of the cocaine dose-response function. MPEP and dizocilpine selectively and significantly attenuated self administration of a low reinforcing dose of cocaine compared to food without evidence of tolerance. CONCLUSIONS: Both MPEP and dizocilpine functioned as partially surmountable antagonists of the reinforcing effects of cocaine. The similar effects of the two drugs raises the possibility that MPEP attenuated the reinforcing effects of cocaine, at least in part, via mGluR5-mediated inhibition of NMDA receptor activity. PMID- 18509622 TI - CB(1) cannabinoid receptor activation dose dependently modulates neuronal activity within caudal but not rostral song control regions of adult zebra finch telencephalon. AB - RATIONALE: CB(1) cannabinoid receptors are distinctly expressed at high density within several regions of zebra finch telencephalon, including those known to be involved in song learning (lMAN and Area X) and production (HVC and RA) because (1) exposure to cannabinoid agonists during developmental periods of auditory and sensory-motor song learning alters song patterns produced later in adulthood and (2) densities of song region expression of CB(1) waxes and wanes during song learning. It is becoming clear that CB(1)-receptor-mediated signaling is important to normal processes of vocal development. MATERIALS AND METHODS: To better understand the mechanisms involved in cannabinoid modulation of vocal behavior, we have investigated the dose-response relationship between systemic cannabinoid exposure and changes in neuronal activity (as indicated by expression of the transcription factor, c-Fos) within telencephalic brain regions, with established involvement in song learning and/or control. RESULTS: In adults, we have found that low doses (0.1 mg/kg) of the cannabinoid agonist WIN-55212-2 decrease neuronal activity (as indicated by densities of c-fos-expressing nuclei) within vocal motor regions of caudal telencephalon (HVC and RA) while higher doses (3 mg/kg) stimulate activity. Both effects were reversed by pretreatment with the CB(1)-selective antagonist rimonabant. Interestingly, no effects of cannabinoid treatment were observed within the rostral song regions lMAN and Area X, despite distinct and dense CB(1) receptor expression within these areas. CONCLUSIONS: Overall, our results demonstrate that, depending on dosage, CB(1) agonism can both inhibit and stimulate neuronal activity within brain regions controlling adult vocal motor output, implicating involvement of multiple CB(1) sensitive neuronal circuits. PMID- 18509623 TI - Effect of +-methamphetamine on path integration learning, novel object recognition, and neurotoxicity in rats. AB - RATIONALE: Methamphetamine (MA) has been implicated in cognitive deficits in humans after chronic use. Animal models of neurotoxic MA exposure reveal persistent damage to monoaminergic systems but few associated cognitive effects. OBJECTIVES: Since questions have been raised about the typical neurotoxic dosing regimen used in animals and whether it adequately models human cumulative drug exposure, these experiments examined two different dosing regimens. MATERIALS AND METHODS: Rats were treated with one of the two regimens: one based on the typical neurotoxic regimen (4 x 10 mg/kg every 2 h) and one based on pharmacokinetic modeling (Cho AK, Melega WP, Kuczenski R, Segal DS Synapse 39:161-166, 2001) designed to better represent accumulating plasma concentrations of MA as seen in human users (24 x 1.67 mg/kg once every 15 min) matched for total daily dose. In two separate experiments, dosing regimens were compared for their effects on markers of neurotoxicity or on behavior. RESULTS: On markers of neurotoxicity, MA showed decreased dopamine (DA) and 5-HT, increased glial fibrillary acidic protein, and increased corticosterone levels regardless of dosing regimen 3 days post-treatment. Behaviorally, MA-treated groups, regardless of dosing regimen, showed hypoactivity, increased initial hyperactivity to a subsequent MA challenge, impaired novel object recognition, impaired learning in a multiple T water maze test of path integration, and no differences on spatial navigation or reference memory in the Morris water maze. After behavioral testing, reductions of DA and 5-HT remained. CONCLUSIONS: MA treatment induces an effect on path integration learning not previously reported. Dosing regimen had no differential effects on behavior or neurotoxicity. PMID- 18509624 TI - A highly sensitive and specific polyclonal antibody-based enzyme-linked immunosorbent assay for detection of antibiotic olaquindox in animal feed samples. AB - The use of olaquindox (OLA) as an additive in animal feedstuffs has been prohibited in the European Union and many other countries. In this study, a highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (ELISA) for determination of OLA in animal feed samples was developed. OLA was activated by N'N-carbonyldiimidazole and coupled with bovine serum albumin (BSA) and ovalbumin (OVA). It was found that the sensitivity and specificity of the two antisera were very similar, with the IC(50) values of 16 ng mL(-1) and 19 ng mL(-1), respectively. Cross-reactivity was less than 35% for four structurally related compounds and no recognition of five other antibiotics was observed. The better antiserum I was selected for further experiments, for example testing stability, solvent effect, accuracy, and precision. The IC(50) value for eight standard curves was in the range 12-18 ng mL(-1) and the LOD at a signal-to-noise ratio of 3 (S/N = 3) was 0.31 +/- 0.11 ng mL(-1). The ELISA tolerated 5% methanol without significant influence on IC(50) value. The recoveries of spiked OLA in five different animal feed types including auxin, pig complex feed, fish complex feed, broiler concentrated feed, and pig premix feed were in the range 88.3 119.0% and the intra-assay relative standard deviation (RSD) was within 4.7-33.5% (n = 3). The ELISA for unspiked feed samples was confirmed by high-performance liquid chromatography (HPLC), with a high correlation coefficient of 0.9862 (n = 5). The proposed ELISA could be a feasible quantitative/screening method for OLA analysis in feed samples with the properties of high sensitivity, specificity, simplicity of sample pretreatment, high sample throughput, and low expense. PMID- 18509626 TI - Was the thrombotic risk of rofecoxib predictable from the French Pharmacovigilance Database before 30 September 2004? AB - OBJECTIVES: Rofecoxib was withdrawn from the market on 30 September 2004 following the results of a randomized controlled trial. Following this sudden decision, several controversies occurred in the literature to determine whether this adverse drug reaction (ADR) could have been detected earlier. The aim of this study was to investigate whether this kind of signal could have been seen using the French Pharmacovigilance Database before this date of rofecoxib withdrawal. METHODS: Using cases registered in the French Pharmacovigilance Database from May 2000 to December 2006, we applied the case-noncase method to "serious" thrombotic ADRs reported with oral formulations of rofecoxib or celecoxib in patients older than 15 years. Cases were all notifications of thrombotic ADRs [World Health Organization Adverse Reaction Terminology (WHO-ART) codes 1300] occurred under coxib (rofecoxib, celecoxib) and noncases all other reports registered in the database (whatever the drug). We calculated a cumulative odds ratio (OR) from 20 May 2000 to 31 December 2006, with a special interest for the period before the 30 September 2004. RESULTS: Among the 50,087 "serious" ADRs registered in the database during this period, 1,127 were thrombotic ones. Rofecoxib exposure was significantly associated with high values of odds ratio (OR) [4.2 (95% CI 1.97-8.61)] for thrombotic ADRs as early as the end of 2001. The values of ADR reporting ORs remained high (3.0-3.5) until 2006. For celecoxib, a significant trend occurred only from September 2004. CONCLUSION: Despite the compulsory limits of the case/noncase methodology, this study found an association between rofecoxib exposure and the occurrence of "serious" thrombotic ADRs as early as the end of the first year of rofecoxib marketing in France. The association between celecoxib and the occurrence of such ADRs appears less clear. Our work also shows the potential use of careful analysis of pharmacovigilance databases (investigating, for example, cumulative values of risk) in the early identification of new ADRs. PMID- 18509627 TI - Perfusion CT compared to H(2) (15)O/O (15)O PET in patients with chronic cervical carotid artery occlusion. AB - INTRODUCTION: The purpose of this study was to compare the results of perfusion computed tomography (PCT) with those of (15)O(2)/H(2) (15)O positron emission tomography (PET) in a subset of Carotid Occlusion Surgery Study (COSS) patients. MATERIALS AND METHODS: Six patients enrolled in the COSS underwent a standard-of care PCT in addition to the (15)O(2)/H(2) (15)O PET study used for selection for extracranial-intracranial bypass surgery. PCT and PET studies were coregistered and then processed separately by different radiologists. Relative measurement of cerebral blood flow (CBF) and oxygen extraction fraction (OEF) were calculated from PET. PCT datasets were processed using different arterial input functions (AIF). Relative PCT and PET CBF values from matching regions of interest were compared using linear regression model to determine the most appropriate arterial input function for PCT. Also, PCT measurements using the most accurate AIF were evaluated for linear regression with respect to relative PET OEF values. RESULTS: The most accurate PCT relative CBF maps with respect to the gold standard PET CBF were obtained when CBF values for each arterial territory are calculated using a dedicated AIF for each territory (R (2) = 0.796, p < 0.001). PCT mean transit time (MTT) is the parameter that showed the best correlation with the count-based PET OEF ratios (R (2) = 0.590, p < 0.001). CONCLUSION: PCT relative CBF compares favorably to PET relative CBF in patients with chronic carotid occlusion when processed using a dedicated AIF for each territory. The PCT MTT parameter correlated best with PET relative OEF. PMID- 18509628 TI - Long-term results with Matrix coils vs. GDC: an angiographic and histopathological comparison. AB - INTRODUCTION: The aim of the study was to compare standard platinum Guglielmi detachable coils (GDC) with coated platinum coils (Matrix; both Boston Scientific, Fremont, CA) regarding handling, complications, occlusion and recanalization rate after 3 and 6A months. METHODS: Aneurysms in the right common carotid artery were created in 25 rabbits. The animals were divided into five groups of five animals each. The animals of group 1 (the control group) received no treatment of the induced aneurysms, the animals of groups 2 and 3 (killed at 3 and 6A months) were treated with standard GDC, and the animals of groups 4 and 5 (killed at 3 and 6A months) were treated with Matrix coils. RESULTS: Histopathological evaluation showed organized thrombus formation and connective tissue with neovascularization around the implanted coils in all the treated groups. The achieved occlusion rates in groups 2 and 3 were identical to those in groups 4 and 5. Thus the long-term results of aneurysm treatment with GDC and Matrix coils show no differences regarding occlusion and recanalization rates. The only noticeable difference was the difference in handling. More force was required to pushing the Matrix coils forward through the microcatheter and there was more friction in coil interaction in the aneurysm. CONCLUSION: The bioactive coating of the Matrix coil produces no significant benefit in achieving higher occlusion and lower recanalization rates, and the coil is more difficult to handle. Future bioactive coils must be shown to produce significantly better long term results than GDC and their ease of handling has to be improved. PMID- 18509629 TI - Improving outcome after positive interim PET in advanced Hodgkin's disease: reality vs expectation. PMID- 18509630 TI - 11C-NMSP/ 18F-FDG microPET to monitor neural stem cell transplantation in a rat model of traumatic brain injury. AB - PURPOSE: To investigate whether (11)C-N-methylspiperone ((11)C-NMSP) microPET could be used for imaging neural stem cells (NSCs) transplantation in a rat model of traumatic brain injury. METHODS: NSCs were induced to express dopamine receptor type 2 (DRD(2)), then confirmed by RT-PCR, Western blotting and immunocytochemistry. Eighteen rats were subjected to focal traumatic brain injury in the right parietal lobe and then assigned randomly to the transplantation group and the control group. NSCs labeled with 5-bromo-2-deoxyuridine (BrdU) were transplanted into the cerebral lesion of the transplantation group. MicroPET scan using (11)C-NMSP and (18)F-FDG were performed to detect the DRD(2) expression of transplanted NSCs and the regional glucose metabolism in the cerebral lesion, respectively. Behavioral neurological function of rats were also tested. RESULTS: Histological analysis identified viable NSCs. Western blotting and immunofluorescence showed high level of NSCs-induced DRD(2) expression. Immunostaining demonstrated high levels of survived BrdU+ and DRD(2)+ donor cells in the cerebral lesion 2 weeks after transplantation. The lesion-to-normal contralateral ratio (L/N ratio) of (11)C-NMSP in the cerebral lesion decreased significantly from 97% to 68% after injury and increased dramatically to 137% 1 day after the transplantation and then decreased gradually. Glucose metabolism showed a decrease of 35% in the cerebral lesion 1 day after injury and recovered to 87% 2 weeks after transplantation. The behavioral neurological function of the transplantation group was significantly improved compared with the control group. CONCLUSIONS: This study verified that (11)C-NMSP microPET can be used to assess the NSCs-induced DRD(2) expression in rat model. PMID- 18509631 TI - Cardiac (123)I-metaiodobenzylguanidine imaging allows early identification of dementia with Lewy bodies during life. AB - PURPOSE: Differential diagnosis between dementia with Lewy bodies (DLB) and other neurodegenerative diseases with cognitive impairment represents a clinical challenge. Due to the overlapping of symptoms, the clinical diagnosis can be modified during the prolonged follow-up of these diseases. The purpose of this study was to assess the ability of cardiac metaiodobenzylguanidine (MIBG) imaging for early identification of DLB. MATERIALS AND METHODS: Since January 2003, all patients with neurodegenerative diseases with cognitive impairment at their first visit at the Memory Unit and clinical criteria of DLB were consecutively recruited and underwent a cardiac (123)I-MIBG study. The heart-to-mediastinum ratio (HMR) and the washout rate (WR) of cardiac MIBG uptake were obtained. RESULTS: Sixty-five patients were included. After a clinical follow-up of 4 years, the progress of the disease procured a definite diagnosis in 44 (68%) patients: 19 DLB, 12 Alzheimer disease (AD), and 13 other neurodegenerative diseases with cognitive impairment. HMR was significantly decreased in DLB with respect to the other neurodegenerative diseases. WR was only significantly different between DLB and AD. The HMR cut off point of 1.36 differentiated DLB from the other dementias with a sensitivity of 94% and a specificity of 96% with an accuracy of 95%. CONCLUSIONS: Cardiac MIBG imaging performed at the time of the first clinical diagnosis of DLB can help early clinical identification or exclusion of this disease. PMID- 18509632 TI - What oncologists need and require from nuclear medicine. PMID- 18509633 TI - Normal differential renal function does not indicate a normal kidney after partial ureteropelvic obstruction and subsequent relief in 2-week-old piglets. AB - PURPOSE: We investigated the functional consequences of relieving ureteric obstruction in young pigs with experimental hydronephrosis (HN) induced by partial unilateral ureteropelvic obstruction. MATERIALS AND METHODS: Three groups of animals were followed from the age of 2 weeks to the age of 14 weeks: Eight animals had severe or grades 3-4 HN throughout the study. Six animals had relief of the obstruction after 4 weeks. Six animals received sham operations at both ages. Morphological and functional examinations were performed at age 6 weeks and again at age 14 weeks and consisted of magnetic resonance imaging (MRI), technetium-diethylenetriaminepentaaceticacid ((99m)Tc-DTPA) renography, renal technetium-dimercaptosuccinicacid ((99m)Tc-DMSA) scintigraphy, and glomerular filtration rate (GFR) measurement. RESULTS: After relief of the partial obstruction, there was reduction of the pelvic diameter and improvement of urinary drainage. Global and relative kidney function was not significantly affected by either obstruction or its relief. Renal (99m)Tc-DMSA scintigraphy showed a change in both the appearance of the kidney and a change in the distribution within kidneys even after relief of obstruction. CONCLUSION: This study shows that partial ureteric obstruction in young pigs may be associated with little effect on global and differential kidney function. However, even after relief of HN, the distribution of (99m)Tc-DMSA in the kidney remains abnormal suggesting that a normal differential renal function may not represent a normal kidney. PMID- 18509634 TI - Use of positron emission tomography for staging, preoperative response assessment and posttherapeutic evaluation in children with Wilms tumour. AB - PURPOSE: To evaluate FDG-PET for staging, grading, preoperative response assessment and posttherapeutic evaluation in children with Wilms tumour (WT). METHODS: In this study, 23 FDG-PET examinations in 12 paediatric patients (female, n = 5; male, n = 7; age, 1-19 years) with WT (primary, n = 9; relapsed, n = 3) were analysed. All patients were examined with conventional imaging methods (CIM) according to the SIOP2001/GPOH trial protocol. Additionally, FDG PET/PET-CT was performed for staging (n = 12), preoperative response assessment (n = 6) and posttherapeutic evaluation (n = 5). Imaging results of FDG-PET and CIM were analysed regarding the accuracy in tumour visualisation, impact on therapeutic management and preoperative response assessment, with clinical follow up and histopathology as the standard of reference. RESULTS: FDG-PET and CIM showed concordant results for staging of primary WT, whereas FDG-PET was superior in 1/3 cases with recurrent WT. Concerning histological differentiation, one case with anaplastic WT had an standard uptake value (SUV) of 12.3, which was remarkably higher than the average SUV in the eight cases with intermediate risk histology. No parameter analysed for PET or CIM was reliably predictive for histological regression or clinical outcome. After completion of therapy, FDG-PET was superior to CIM in 2/5 cases in detecting residual disease with therapeutic relevance. CONCLUSION: FDG-PET does not provide additional information to the traditional imaging work-up for staging WT patients, preoperative response assessment and clinical outcome. FDG-PET was advantageous in ruling out residual disease after completion of first line treatment and in pretherapeutic staging of relapse patients. Furthermore, there seems to be a good correlation of initial SUV and histological differentiation. PMID- 18509635 TI - Tetrafluorophenolate of HBED-CC: a versatile conjugation agent for 68Ga-labeled small recombinant antibodies. AB - PURPOSE: The success of (68)Ga-labeled peptides for positron emission tomography of neuroendocrine tumors is mainly depending on the complex chemistry of this radioisotope. 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), the chelator of choice has however limitations if its application is expanded to heat-sensitive proteins. Recombinant antibodies like single chain Fv or diabodies belong to this class of proteins. They are suited to provide imaging contrast despite the short-lived (68)Ga because of their rapid blood clearances and nanomolar affinities. The heterobifunctional agent N,N'-bis[2-hydroxy-5 (carboxyethyl)benzyl]ethylenediamine-N,N'-diacetic acid (HBED-CC) was chosen as an alternative ligand because this agent is complexing [(68)Ga]Ga(3+) much faster than DOTA at ambient temperatures. MATERIALS AND METHODS: A versatile technology for HBED-CC conjugation of proteins and (68)Ga-labeling has been developed. This included HBED-CC-tetrafluorophenol (TFP) ester synthesis, coupling to the antibody at various pH and complexation reactions performed in 4-(2-hydroxyethyl) 1-piperazineethanesulfonic acid (HEPES) buffer under different conditions. RESULTS: The synthesis of the monoreactive 2,3,5,6-tetrafluorophenolate of HBED CC at a carboxyl group not participating in complex formation used [Fe(HBED-CC)]( ) for ester formation. The removal of Fe(3+) from purified (HBED-CC)TFP ester was achieved with RP(18) cartridge technology. The conjugation chemistry was performed with mAb425 which binds to the epidermal growth factor receptor (EGFR). This protein was used for optimizing purposes only. The influence of complexation parameters like temperature, pH, reaction time, and HBED-CC/antibody ratio on the biological activity of this model antibody was investigated. Furthermore, the outcome of this labeling procedure on the biological activity of a recombinant diabody (50 kDa) was studied. CONCLUSION: It is known that small HBED-CC/antibody ratios are prerequisites for minimal interference of labels with antigen-binding domains. Here, the coupling of about one HBED-CC per antibody proved to be sufficient for efficient (68)Ga labeling, pointing to the successful application of (68)Ga for molecular imaging with small recombinant proteins. PMID- 18509636 TI - Macrocyclic chelator-coupled gastrin-based radiopharmaceuticals for targeting of gastrin receptor-expressing tumours. AB - PURPOSE: Diethylenetriamine-pentaacetic acid (DTPA)-coupled minigastrins are unsuitable for therapeutic application with the available beta-emitting radiometals due to low complex stability. Low tumour-to-kidney ratio of the known radiopharmaceuticals is further limiting their potency. We used macrocyclic chelators for coupling to increase complex stability, modified the peptide sequence to enhance radiolytic stability and studied tumour-to-kidney ratio and metabolic stability using (111)In-labelled derivatives. METHODS: Gastrin derivatives with decreasing numbers of glutamic acids were synthesised using (111)In as surrogate for therapeutic radiometals for in vitro and in vivo studies. Gastrin receptor affinities of the (nat)In-metallated compounds were determined by receptor autoradiography using (125)I-CCK as radioligand. Internalisation was evaluated in AR4-2J cells. Enzymatic stability was determined by incubating the (111)In-labelled peptides in human serum. Biodistribution was performed in AR4-2J-bearing Lewis rats. RESULTS: IC(50) values of the (nat)In metallated gastrin derivatives vary between 1.2 and 4.8 nmol/L for all methionine containing derivatives. Replacement of methionine by norleucine, isoleucine, methionine-sulfoxide and methionine-sulfone resulted in significant decrease of receptor affinity (IC(50) between 9.9 and 1,195 nmol/L). All cholecystokinin receptor affinities were >100 nmol/L. All (111)In-labelled radiopeptides showed receptor-specific internalisation. Serum mean-life times varied between 2.0 and 72.6 h, positively correlating with the number of Glu residues. All (111)In labelled macrocyclic chelator conjugates showed higher tumour-to-kidney ratios after 24 h (0.37-0.99) compared to (111)In-DTPA-minigastrin 0 (0.05). Tumour wash out between 4 and 24 h was low. Imaging studies confirmed receptor-specific blocking of the tumour uptake. CONCLUSIONS: Reducing the number of glutamates increased tumour-to-kidney ratio but resulted in lower metabolic stability. The properties of the macrocyclic chelator-bearing derivatives make them potentially suitable for clinical purposes. PMID- 18509637 TI - FDG uptake, a surrogate of tumour hypoxia? AB - INTRODUCTION: Tumour hyperglycolysis is driven by activation of hypoxia-inducible factor-1 (HIF-1) through tumour hypoxia. Accordingly, the degree of 2-fluro-2 deoxy-D: -glucose (FDG) uptake by tumours might indirectly reflect the level of hypoxia, obviating the need for more specific radiopharmaceuticals for hypoxia imaging. DISCUSSION: In this paper, available data on the relationship between hypoxia and FDG uptake by tumour tissue in vitro and in vivo are reviewed. In pre clinical in vitro studies, acute hypoxia was consistently shown to increase FDG uptake by normal and tumour cells within a couple of hours after onset with mobilisation or modification of glucose transporters optimising glucose uptake, followed by a delayed response with increased rates of transcription of GLUT mRNA. In pre-clinical imaging studies on chronic hypoxia that compared FDG uptake by tumours grown in rat or mice to uptake by FMISO, the pattern of normoxic and hypoxic regions within the human tumour xenografts, as imaged by FMISO, largely correlated with glucose metabolism although minor locoregional differences could not be excluded. In the clinical setting, data are limited and discordant. CONCLUSION: Further evaluation of FDG uptake by various tumour types in relation to intrinsic and bioreductive markers of hypoxia and response to radiotherapy or hypoxia-dependent drugs is needed to fully assess its application as a marker of hypoxia in the clinical setting. PMID- 18509638 TI - Multicentre use of a porous tantalum monoblock acetabular component. AB - The purpose of this study was to evaluate the minimum five-year prospective results from the multicentre use of a porous tantalum monoblock acetabular component for primary total hip arthroplasty (THA). A multicentre study was performed in 253 consecutive primary THAs in three separate surgical centres. All patients underwent identical postoperative protocols including radiological and clinical evaluation. The average preoperative total HHS score was 44.0 +/- 13.8 and increased at one-year follow-up to 95.2 +/- 4.8 (p <0.05), remaining constant through the five-year follow-up at 97.0 +/- 6.2 (p < 0.05). There was no radiographic evidence of gross polyethylene wear, progressive radiolucencies, osteolytic lesions, acetabular fracture, or component subsidence. From these results, we can recommend the continued use of this material for acetabular components in primary THA and that further review of the current multicentre population is warranted to determine the long-term durability of the acetabular composite. PMID- 18509639 TI - Is extreme flexion of the knee after total knee replacement a prerequisite for patient satisfaction? AB - The focus of this study was to evaluate the functional result and to specifically ascertain whether the absence of the ability to squat and sit cross-legged altered the patient's satisfaction level after a successful standard total knee replacement. Squatting and sitting cross-legged are common practices in Asia. These activities are not possible following standard total knee replacement. Patients were followed-up for a minimum of 12 months post surgery. Their level of satisfaction was assessed using a Likert scale. The Knee Society Score (KSS) was used to assess range of motion and function of the knee. Twenty-one out of 25 patients were satisfied with the surgical result in spite of an inability to squat. Deep knee flexion may not be an essential prerequisite for patient satisfaction after total knee replacement, even in a population where squatting and sitting cross-legged are part of the normal lifestyle. PMID- 18509640 TI - Acquired long QT syndrome and sudden cardiac death due to secondary hemochromatosis with multitransfusions for severe aplastic anemia. PMID- 18509641 TI - Fluorine-18 fluorodeoxyglucose positron emission tomography in mature T-cell and natural killer cell malignancies. AB - Fluorine-18 fluorodeoxyglucose (FDG)-positron emission tomography (PET) is useful in Hodgkin and B-cell lymphomas. Few data exist on T-cell and natural killer (NK) cell lymphomas. Thirty consecutive T-cell and NK-cell lymphomas were investigated with PET-computerized tomography (CT). In 12 NK-cell lymphomas, all nasal/extranasal lesions were FDG-avid. In nasal/maxillary areas, FDG-avid tumours were consistently more localised than on CT, suggesting that soft tissue masses on CT were partly due to inflammation. These findings have important implications in radiotherapy planning. In two NK-cell lymphomas, PET did not detect morphologically occult marrow infiltration uncovered by in situ hybridisation for Epstein-Barr-virus-encoded small RNA. In angioimmunoblastic lymphoma (n = 7), peripheral T-cell lymphoma, unspecified (PTCL-U, n = 4) and anaplastic large cell lymphoma (ALCL, n = 3), involved nodal/extranodal sites shown on CT and/or biopsy were concordantly PET-positive. In one PTCL-U, PET detected FDG-avid marrow infiltrations not shown on biopsies. In contrast, cutaneous ALCL (n = 1) and mycosis fungoides (n = 2) showed minimal FDG uptake. In one case of T-cell large granular lymphocyte leukaemia, marrow, nodal and bowel infiltrations were not FDG-avid. PET maximum standardised uptake value did not correlate with clinicopathological features and prognosis. These observations defined the pre-treatment value of PET-CT in T-cell and NK-cell lymphomas. The post-treatment role requires further studies. PMID- 18509642 TI - Efficacy, pharmacokinetics, tisssue distribution, and metabolism of the Myc-Max disruptor, 10058-F4 [Z,E]-5-[4-ethylbenzylidine]-2-thioxothiazolidin-4-one, in mice. AB - OBJECTIVES: c-Myc is commonly activated in many human tumors and is functionally important in cellular proliferation, differentiation, apoptosis and cell cycle progression. The activity of c-Myc requires noncovalent interaction with its client protein Max. In vitro studies indicate the thioxothiazolidinone, 10058-F4, inhibits c-Myc/Max dimerization. In this study, we report the efficacy, pharmacokinetics and metabolism of this novel protein-protein disruptor in mice. METHODS: SCID mice bearing DU145 or PC-3 human prostate cancer xenografts were treated with either 20 or 30 mg/kg 10058-F4 on a qdx5 schedule for 2 weeks for efficacy studies. For pharmacokinetics and metabolism studies, mice bearing PC-3 or DU145 xenografts were treated with 20 mg/kg of 10058-F4 i.v. Plasma and tissues were collected 5-1440 min after dosing. The concentration of 10058-F4 in plasma and tissues was determined by HPLC, and metabolites were characterized by LC-MS/MS. RESULTS: Following a single iv dose, peak plasma 10058-F4 concentrations of approximately 300 muM were seen at 5 min and declined to below the detection limit at 360 min. Plasma concentration versus time data were best approximated by a two-compartment, open, linear model. The highest tissue concentrations of 10058-F4 were found in fat, lung, liver, and kidney. Peak tumor concentrations of 10058-F4 were at least tenfold lower than peak plasma concentrations. Eight metabolites of 10058-F4 were identified in plasma, liver, and kidney. The terminal half-life of 10058-F4 was approximately 1 h, and the volume of distribution was >200 ml/kg. No significant inhibition of tumor growth was seen after i.v. treatment of mice with either 20 or 30 mg/kg 10058-F4. CONCLUSION: The lack of significant antitumor activity of 10058-F4 in tumor bearing mice may have resulted from its rapid metabolism and low concentration in tumors. PMID- 18509643 TI - Toxicity and toxicokinetics of the cyclin-dependent kinase inhibitor AG-024322 in cynomolgus monkeys following intravenous infusion. AB - PURPOSE: Cyclin-dependent kinases (CDKs) play a significant role in the control of cell-cycle progression and exhibit aberrant regulation in various neoplastic diseases. AG-024322 is a potent inhibitor of CDK1, CDK2, and CDK4 that produces cell-cycle arrest and antitumor activity in preclinical models. This study evaluated the toxicity of AG-024322 when given by intravenous (IV) infusion to cynomolgus monkeys, including reversibility of effects. METHODS: Male and female monkeys received AG-024322 by 30-min IV infusion once daily for 5 days at doses of 2, 6, and 10 mg/kg (24, 72, and 120 mg/m(2), respectively). Controls received vehicle alone which was aqueous 5% dextrose, pH 3.8. Three animals/sex/group were necropsied on day 6, and two animals/sex/group at 6 and 10 mg/kg were necropsied on day 22 (reversal cohort). Doses were based upon the results of a dose range finding study in monkeys; decreased white blood cells occurred at > or =3 mg/kg and 12 mg/kg produced central nervous system effects and was above the maximum tolerated dose. RESULTS: No deaths occurred and clinical signs of toxicity, including swelling at the IV administration site, were seen at > or =6 mg/kg. AG 024322 at > or =6 mg/kg produced pancytic bone marrow hypocellularity, lymphoid depletion, and vascular injury at the injection site. Renal tubular degeneration occurred at 10 mg/kg. These changes were either reversible or in a process of repair following the 17-day recovery period. Hematology changes included decreases in reticulocytes and/or granulocytes at > or =6 mg/kg, which were reversible and consistent with changes in the bone marrow. Lymphoid and bone marrow depletion are consistent with pharmacologic inhibition of CDKs by AG 024322 and were expected findings. On day 22, vacuolar degeneration of pancreatic acinar cells with increased serum amylase and lipase levels occurred in one female at 10 mg/kg. Neither sex-related differences in toxicokinetics nor plasma accumulation over 5 days of dosing were seen. Terminal phase overall mean half life on day 5 ranged from 6.69 to 8.87 h (across dose levels) and was not dose dependent. CONCLUSION: The no-adverse-effect dose of AG-024322 was 2 mg/kg and associated with overall mean plasma AUC(0-24.5) of 2.11 microg h/mL. PMID- 18509644 TI - Targeting MET transcription as a therapeutic strategy in multiple myeloma. AB - Multiple myeloma (MM) is an incurable indolent malignancy with an average lifespan of 3 years, underscoring the need for new therapies. Studies have shown that the receptor MET and its ligand hepatocyte growth factor play an important role in proliferation, migration, adhesion, and survival of MM cells. Hence, an effective way to decrease MET receptor may act as a viable therapeutic option. Since MET mRNA and protein have short half-lives, we hypothesized that transcription inhibitor will reduce MET transcript and protein levels and this will lead to cell death. Pharmacological (flavopiridol) and molecular (shRNA) transcription inhibitor were used to impede formation of MET transcripts. The diminution of global RNA synthesis with flavopiridol was related to phosphorylation status of Ser residues (r (2) = 0.90 and 0.92 for Ser2 and Ser5) on the C-terminal-domain of RNA polymerase II. This was accompanied with a time dependent decrease in MET transcript, which reached to less than 30% (1 microM) and 10% (3 microM) by 24 h. This decline in transcript level was directly associated with a reduction in MET protein level (r (2) = 0.82) and resulted in cell death. Assessment of MET in MM survival was done by using shRNA targeted towards MET. When cells were infected with shRNA viral construct, there was increased cell death with a decline in MET transcript and protein. Taken together, our study demonstrates that MET plays a critical role in the survival and removal or lowering of MET by flavopiridol or shRNA results in the demise of MM cells. PMID- 18509645 TI - A dose-ranging study of the pharmacokinetics and pharmacodynamics of the selective apoptotic antineoplastic drug (SAAND), OSI-461, in patients with advanced cancer, in the fasted and fed state. AB - PURPOSE: To evaluate the safety, pharmacokinetics and determine the recommended dose of the selective apoptotic antineoplastic drug, OSI-461 administered on a twice-daily regimen to patients with advanced solid malignancies. METHODS: In this phase I trial, 33 patients were treated with OSI-461 doses ranging from 400 to 1,200 mg given twice daily in 4-week cycles. Pharmacokinetic studies were performed to characterize the plasma disposition of OSI-461 and the effect of food intake on OSI-461 absorption. Secondary biomarker studies were performed to assess the biologic activity of OSI-461 including the measurement of pGSK-3beta, a PKG substrate, and pharmacogenetic studies to identify polymorphisms of CYP3A that influence drug metabolism and of ABCG2, involved in drug resistance. RESULTS: Thirty-three patients were treated with 86 courses of OSI-461. The dose limiting toxicities were grade 3 abdominal pain, found in one patient at the 1,000 mg BID fed dose level and all patients at the 1,200 mg BID fed dose level. There was also one episode each of grade 3 fatigue and grade 3 constipation at the 1,000 and 1,200 mg BID fed dose levels, respectively. Other common toxicities included mild to moderate fatigue, nausea, anorexia and mild elevation in bilirubin. Pharmacokinetic studies of OSI-461 revealed approximately a twofold increase in AUC(0-24) when OSI-461 was administered with food. An increase in pGSK-3beta post-dose was seen in the majority of patients and was greater at higher dose levels. No patients exhibited CYP3A4 polymorphisms, while 100% of patients were found to have the CYP3A5*3/CYP3A5*3 polymorphism. Two known polymorphisms of the ABCG2 gene, G34 --> A34 and C421 --> A421, occurred at frequencies of 11.76 and 29%, respectively. CONCLUSIONS: Toxicity and pharmacodynamic data show that the recommended oral dose of OSI-461 is 800 mg twice daily administered with food. The drug appears to be well-tolerated, and overall bioavailability appears to be markedly increased when the drug is administered with food. These results support further disease-directed evaluations of OSI-461 at a dose of 800 mg BID in combination with other chemotherapeutic agents. PMID- 18509646 TI - NOX enzymes as novel targets for drug development. AB - The members of the NOX/DUOX family of NADPH oxidases mediate such physiologic functions as host defense, cell signaling, and thyroid hormone biosynthesis through the generation of reactive oxygen species (ROS), including superoxide anion and hydrogen peroxide. Moreover, ROS are involved in a broad range of fundamental biochemical and cellular processes, and data accumulated in recent years indicate that the NOX enzymes comprise one of the most important biological sources of ROS. Given the high biochemical reactivity of ROS, it is not surprising that they have been implicated in a wide variety of pathologies and diseases. Prominent among the settings that feature ROS-mediated tissue injury are disorders associated with inflammation, aging, and progressive degenerative changes in cells and organ systems, and it appears that essentially no organ system is exempt. Among the disorders currently believed to be mediated at least in part by NOX-derived ROS are hypertension, aortic aneurysm, myocardial infarction (and other ischemia-reperfusion disorders), pulmonary fibrosis and hypertension, amyotropic lateral sclerosis, Alzheimer's disease, Parkinson's disease, ischemic stroke, diabetic nephropathy, and renal cell carcinoma. Several small-molecule and peptide inhibitors of the NOX enzymes have been useful in experimental studies, but issues of specificity, potency, and toxicity militate against any of the existing published compounds as candidates for drug development. Given the broad array of disease targets documented in recent work, the time is here for vigorous efforts to develop clinically useful inhibitors of the NOX enzymes. As most (though not all) NOX-related diseases appear to be mediated by a single member of the NOX family, agents with isoform specificity will be preferred, although broadly active NOX inhibitors may prove to be useful in some settings. PMID- 18509647 TI - Genetics and immunopathology of chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a primary immunodeficiency syndrome characterized by a greatly increased susceptibility to severe fungal and bacterial infections. CGD results from a failure of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme in the patient's phagocytes to produce superoxide. It is caused by mutations in any of four genes that encode the components of the NADPH oxidase. Investigation of CGD patients has identified the different subunits and the genes encoding them. Study of rare CGD variants has highlighted sequences involved in the structural stability of affected components or has provided valuable insights into their function in the oxidase activation mechanism. Functional and molecular CGD diagnosis tests are discussed in this review. Long-term antibiotic prophylaxis has been essential in fighting infections associated with CGD, but approaches based on hematopoietic stem cell transplantation and gene therapy offer great hope for the near future. PMID- 18509648 TI - Hyperinflammation in chronic granulomatous disease and anti-inflammatory role of the phagocyte NADPH oxidase. AB - Chronic granulomatous disease (CGD) is an immunodeficiency caused by the lack of the superoxide-producing phagocyte nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. However, CGD patients not only suffer from recurrent infections, but also present with inflammatory, non-infectious conditions. Among the latter, granulomas figure prominently, which gave the name to the disease, and colitis, which is frequent and leads to a substantial morbidity. In this paper, we systematically review the inflammatory lesions in different organs of CGD patients and compare them to observations in CGD mouse models. In addition to the more classical inflammatory lesions, CGD patients and their relatives have increased frequency of autoimmune diseases, and CGD mice are arthritis-prone. Possible mechanisms involved in CGD hyperinflammation include decreased degradation of phagocytosed material, redox-dependent termination of proinflammatory mediators and/or signaling, as well as redox-dependent cross-talk between phagocytes and lymphocytes (e.g. defective tryptophan catabolism). As a conclusion from this review, we propose the existence of ROS high and ROS low inflammatory responses, which are triggered as a function of the level of reactive oxygen species and have specific characteristics in terms of physiology and pathophysiology. PMID- 18509649 TI - A branching process for the early spread of a transposable element in a diploid population. AB - Transposable elements (TEs) face significant challenges upon transfer into a new host population, invariably beginning their invasion with only a single element. The fate of this element is a product of its internal properties, the population dynamics of the host species, and genetic drift. We present a continuous-time multi-type branching process to model the early stages of TE spread. The model incorporates seasonal population size changes and is applicable to diploid hosts for prevalences up to 10%. We reproduce standard results of TE population dynamics and show that population growth may have a greater influence on reducing TE loss probability than a transpositional burst. These results are applied to the planned use of a TE to drive an antimalarial gene into an Anopheles gambiae population. The model favors a transgenic release immediately following the dry season when the An. gambiae population begins to grow. Increasing the number of transgenic hosts released has the greatest influence on reducing the probability of TE loss. Following release, the rate at which the TE increases its proportion in the population is most sensitive to its replicative transposition rate. The model recommends a replicative transposition rate greater than 0.1 per TE per generation to satisfy public health goals. PMID- 18509650 TI - Stochastic properties of generalised Yule models, with biodiversity applications. AB - The Yule model is a widely used speciation model in evolutionary biology. Despite its simplicity many aspects of the Yule model have not been explored mathematically. In this paper, we formalise two analytic approaches for obtaining probability densities of individual branch lengths of phylogenetic trees generated by the Yule model. These methods are flexible and permit various aspects of the trees produced by Yule models to be investigated. One of our methods is applicable to a broader class of evolutionary processes, namely the Bellman-Harris models. Our methods have many practical applications including biodiversity and conservation related problems. In this setting the methods can be used to characterise the expected rate of biodiversity loss for Yule trees, as well as the expected gain of including the phylogeny in conservation management. We briefly explore these applications. PMID- 18509651 TI - Effect of prayer on osteoarthritis and osteoporosis: any difference between men and women? PMID- 18509652 TI - Leukocytapheresis for the treatment of refractory Henoch-Schonlein purpura resistant to both prednisolone and intravenous immunoglobulin therapy. AB - Henoch-Schonlein purpura (HSP) is a childhood disorder, which is the vasculitis of systemic small vessels of unknown etiology. We encountered the dramatic efficacy of leukocytapheresis in a Japanese girl with refractory HSP resistant to combined prednisolone plus intravenous immunoglobulin therapy administration. PMID- 18509653 TI - Comparative physiological and molecular responses of a common aromatic indica rice cultivar to high salinity with non-aromatic indica rice cultivars. AB - In an attempt to understand the molecular basis of salt-stress response in the aromatic rice Gobindobhog, a comprehensive analysis encompassing physiological or biochemical assays and gene expression studies under high salt (200 mM NaCl) supply regimes were initiated and compared with a salt-sensitive (M-1-48) and salt-tolerant (Nonabokra) rice. The detrimental effects of salinity stress were the most pronounced in Gobindobhog, as reflected by the maximally increased root to shoot ratio, the highest chlorophyll degeneration, the highest foliar concentration of Na(+) ions and peroxide content, with their maximum increment after salt treatment. The amplification of oxidative damages was further stimulated by the accumulation of putrescine and lipid peroxidation-derived toxic degradation products (increased malondialdehyde and lipoxygenase activity), which were comparable in M-1-48 and Gobindobhog. Antioxidants like anthocyanin and particularly cysteine and the osmolytes like reducing sugar, proline and polyamines (spermidine and spermine) showed the highest level in Nonabokra. While the inhibition of catalase activity occurred in all the varieties following salt stress, the maximum induction in guaiacol peroxidase activity, elevated cysteine and proline levels in Gobindobhog probably constituted the detoxification mechanism obligatory for its survival. Intensification of the aroma content with salt treatment was markedly noted in Gobindobhog. A very low abundance of Rab16A/SamDC transcript and the corresponding proteins were observed both in M-1 48 and Gobindobhog, induced only after salt-stress, whereas they were constitutively expressed in Nonabokra. Thus, our data reflect Gobindobhog as a salt-sensitive cultivar, susceptible to high-stress-induced growth-inhibition, ion imbalances, membrane/oxidative damages with lower expression of stress tolerant genes. PMID- 18509654 TI - Mammalian pro-apoptotic bax gene enhances tobacco resistance to pathogens. AB - Emerging evidence suggests that plants and animals may share certain biochemical commonalities in apoptosis, or programmed cell death (PCD) pathways, though plants lack key animal apoptosis related genes. In plants, PCD has many important functions including a role in immunity and resistance to pathogen infection. In this study, a rice phenylalanine ammonia-lyase promoter is used to regulate the expression of the mouse pro-apoptotic bax gene in transgenic tobacco plants. Ectopic expression of the bax negatively affects the growth of transgenic plants. Nonetheless, results show that the bax transgene is induced upon infection by plant pathogens and accumulation of Bax is observed by Western blot analysis. By estimating and measuring the extent of cell death, release of active oxygen species, and accumulation defense-associated gene transcripts, it is shown that bax transgenic plants mount a more robust cell death response compared to control plants. The bax transgenic tobacco plants are also more resistant to infection by Phytophthora parasitica and Ralstonia solanacearum, but have no obvious resistance to tobacco mosaic virus. These results substantiate past studies and illustrate the powerful effects mammalian bax genes may have on plant development and disease resistance. PMID- 18509655 TI - Preoperative staging of biliary carcinoma using 18F-fluorodeoxyglucose PET: prospective comparison with PET+CT, MDCT and histopathology. AB - The aim of this study was to evaluate the value of positron emission tomography with (18)F-labeled fluorodeoxyglucose (FDG-PET) as a preoperative diagnostic investigation in patients with biliary carcinoma. Seventy-two patients with potentially resectable biliary carcinoma underwent preoperative multidetector-row computed tomography (MDCT) and FDG-PET. Both diagnoses were compared with subsequent histopathology and follow-up results. In 64 lesions with biliary carcinoma, 57 (89%) revealed an intense focal accumulation on FDG-PET and were interpreted as malignant. On the other hand, eight benign lesions did not show any specific accumulation. Detection rate of FDG-PET in the nodular type of the tumour (96% or 27/28) was superior to that of the infiltrating type (74% or 17/23) (p = 0.037). For the evaluation of lymph node metastasis, the overall accuracy was 69% (35/51) in both FDG-PET and MDCT: FDG-PET had a lower sensitivity (33% vs. 57%) and a higher specificity (97% vs. 79%) than MDCT, although the values were not significantly different. FDG-PET revealed all six lesions of distant metastases in six patients including two lesions missed by MDCT. FDG-PET has high detectability of biliary malignancies. Like MDCT, FDG-PET offers only modest accuracy for regional lymph node staging, but it may reveal distant metastases missed by MDCT. PMID- 18509656 TI - Demonstration of the Adamkiewicz artery in patients with descending or thoracoabdominal aortic aneurysm: optimization of contrast-medium application for 64-detector-row CT angiography. AB - Our aim was to prospectively investigate the contrast agent concentration and the infusion rate to obtain optimal images of the Adamkiewicz artery by 64-row MDCT. Eighty patients were divided into four groups based on the following protocol: A, 100 mL of 300 mg I/mL at 5.0 mL/s; B, 100 mL of 350 mg I/mL at 5.0 mL/s; C, 100 mL of 300 mg I/mL at 3.5 mL/s; D, 100 mL of 350 mg I/mL at 3.5 mL/s. Quantitative evaluation was performed by calculating mean aortic attenuation value from T5 to L3 level. Visual evaluation of the Adamkiewicz artery was also performed. In a quantitative evaluation, the rapid injection (A, 473.4 +/- 82.6 HU; B, 506.7 +/- 71.9 HU) was significantly superior to the slow injection (C, 371.3 +/- 65.1 HU; D, 391.5 +/- 60.8 HU). In a visual evaluation, the Adamkiewicz artery was assessable in 15 of 20 (75%), 16 of 20 (80%), 9 of 20 (45%), and 10 of 20 (50%) patients in groups A, B, C, and D, respectively. The injection protocol of 1.75 g iodine/s provides both the adequate aortic attenuation of more than 450 HU to assess the Adamkiewicz artery and the best visualization of the Adamkiewicz artery in 64-row MDCT. PMID- 18509657 TI - Image quality improvement of composed whole-spine MR images by applying a modified homomorphic filter--first results in cases of multiple myeloma. AB - To establish a modified homomorphic filter (BiFiC) for post-processing of composed MR images in clinical routine and to evaluate it in special regards to image quality and diagnostic safety. Twenty-three whole-spine examinations were post-processed with the filter. Qualitative image evaluation included documentation of lesions and their visualization at original and post-processed images. Variations of signal intensities were calculated pixel by pixel and visualized by color-coded maps. Quantitative data evaluation was conducted by region-by-region analysis with standardized regions of interests. The BiFiC filter could be implemented successfully on the scanner's software platform and used within clinical routine. Color-coded maps could demonstrate that the BiFiC filter improves the signal uniformity in all cases, including images with metallic artifacts caused by implants. The subjective image quality of the post processed images was improved in 22 out of the 23 MR examinations; in one case it was rated as equal. All pathologies were visualized on post-processed images without the need of additional contrast adjustments. The implemented BiFiC filter significantly improves image signal homogeneity. The algorithm can consequently be integrated into clinical routine as an automatic image post-processing step. PMID- 18509658 TI - Multi-contrast, isotropic, single-slab 3D MR imaging in multiple sclerosis. AB - To describe signal and contrast properties of an isotropic, single-slab 3D dataset [double inversion-recovery (DIR), fluid-attenuated inversion recovery (FLAIR), T2, and T1-weighted magnetization prepared rapid acquisition gradient echo (MPRAGE)] and to evaluate its performance in detecting multiple sclerosis (MS) brain lesions compared to 2D T2-weighted spin-echo (T2SE). All single-slab 3D sequences and 2D-T2SE were acquired in 16 MS patients and 9 age-matched healthy controls. Lesions were scored independently by two raters and characterized anatomically. Two-tailed Bonferroni-corrected Student's t-tests were used to detect differences in lesion detection between the various sequences per anatomical area after log-transformation. In general, signal and contrast properties of the 3D sequences enabled improved detection of MS brain lesions compared to 2D-T2SE. Specifically, 3D-DIR showed the highest detection of intracortical and mixed WM-GM lesions, whereas 3D-FLAIR showed the highest total number of WM lesions. Both 3D-DIR and 3D-FLAIR showed the highest number of infratentorial lesions. 3D-T2 and 3D-MPRAGE did not improve lesion detection compared to 2D-T2SE. Multi-contrast, isotropic, single-slab 3D MRI allowed an improved detection of both GM and WM lesions compared to 2D-T2SE. A selection of single-slab 3D contrasts, for example, 3D-FLAIR and 3D-DIR, could replace 2D sequences in the radiological practice. PMID- 18509659 TI - The effect of indomethacin on systemic and renal hemodynamics in neonatal piglets during experimental endotoxemia. AB - Systemic and renal hemodynamics are affected by prostaglandin production during endotoxemia. To study indomethacin effects on endotoxinemia in a neonatal piglet model, sixteen 7-10 day old piglets were anesthetized, ventilated, and catheterized. Mean arterial pressure (MAP), heart rate (HR), and urine output were continuously monitored. Endotoxin (0.06 mcg/kg) was injected after baseline measurements. We studied two groups with either endotoxinemia alone (n = 7) or an additional indomethacin infusion (0.2 mg/kg per h, n = 9). HR, MAP, renal blood flow (RBF), systemic and renal vascular resistance (SVR, RVR), cardiac index (CI), and glomerular filtration rate (GFR), were obtained at baseline, at 1, 2 and 3 h. We observed a drop in CI and an increase in SVR and HR within 3 h of endotoxinemia, while MAP remained unchanged. These effects were prevented by indomethacin. RVR was not altered significantly. Endotoxinemia triggered a drop of RBF in both control (P < 0.01) and intervention group (P < 0.05). In the intervention group, drop of GFR, urine volume, and paraaminohippuric acid clearance were apparent signs of nephrotoxicity (P < 0.01, <0.05, and <0.01). In conclusion, indomethacin maintains hemodynamic parameters during endotoxinemia at the expense of nephrotoxicity. We speculate that indomethacin counteracts the renoprotective effect of prostaglandins. PMID- 18509660 TI - Characteristic findings on defecography according to reconstruction method and defecatory disorder following sphincter-saving surgery for rectal cancer. AB - BACKGROUND AND AIMS: This study used postoperative defecography to characterize morphological features of defecatory disorders in patients following rectal resection. We also evaluated differences in dynamic defecatory condition depending on reconstruction methods for sphincter-saving surgery. MATERIALS AND METHODS: Subjects comprised 62 patients (male/female, 41/21; mean age, 61 years) who underwent defecography after sphincter-saving surgery for rectal cancer. Semisolid barium (100 ml) was introduced into the rectum, and images were taken in a sitting position. Characteristic dynamic findings in defecography were evaluated according to operative methods and were compared with symptoms of defecatory disorders. RESULTS: Defecographic findings closely associated with postoperative defecatory disorder were as follows: (1) low volume of neorectum in patients with worse incontinence grade (p < 0.05), (2) low evacuation fraction in patients with significantly impaired function such as soiling, urgency, and worsened incontinence score (p < 0.05), (3) minor alteration of anorectal angle at evacuation in patients with major soiling and worsened incontinence score (p < 0.05), and (4) barium shadow in the anal canal at rest in patients with urgency (p < 0.05). By reconstruction method, the J-pouch displayed a larger volume than straight anastomosis but a significantly wider anorectal angle than high anterior resection (HAR). Side-to-end anastomosis offered a moderate volume and a sharp anorectal angle as in HAR. CONCLUSIONS: Defecography is useful for visualizing and characterizing defecatory disorders following rectal resection. Based on defecography, J-pouch reconstruction offers advantageous volume, while side-to end anastomosis provides a more acute anorectal angle for patients who have received rectal resection with low anastomosis. A new reconstruction method offering both advantages was discussed. PMID- 18509661 TI - [Prevalence of musculoskeletal complaints and self-reported joint osteoarthritis in the population of Herne : a telephone survey]. AB - BACKGROUND: Musculoskeletal pain is among the most common symptoms in the population and osteoarthritis is the most important underlying disease. Due to demographic changes, an increase in problems with arthritis is to be expected. To assess the impact on individuals affected and the community, data on the prevalence of musculoskeletal pain and osteoarthritis are essential. METHODS: We performed a telephone survey in a sample of 1,270 inhabitants of the City of Herne, Germany, aged 40 years and older. Participants were asked to give their experience on musculoskeletal pain at the time of the questionnaire and during the last 4 weeks and 12 months. Further questions were whether the knee or hip were the site of most severe pain and if osteoarthritis was ever diagnosed by a physician. Standardized prevalences are reported according to age. RESULTS: A total of 862 (67.9%) persons participated and participants were significantly older and more often female. Musculoskeletal pain on the day of the questionnaire, during the past 4 weeks and the past 12 months was reported in 37.4%, 53.0% and 60.0%, respectively. The knee and hip were predominantly affected in 35.9% and 16.1%, respectively and 26.2% reported that a physician had previously diagnosed osteoarthritis. The prevalences were related to age and gender. CONCLUSIONS: Musculoskeletal pain and osteoarthritis affect an important part of the adult urban population. We found associations between self-reported musculoskeletal pain, osteoarthritis, age and gender. Further studies should evaluate the interference of pain with activities of daily living as well as the use of health services by affected patients. PMID- 18509662 TI - Close association of water channel AQP1 with amyloid-beta deposition in Alzheimer disease brains. AB - Aquaporin-1 (AQP1), a membrane water channel protein, is expressed exclusively in the choroid plexus epithelium in the central nervous system under physiological conditions. However, AQP1 expression is enhanced in reactive astrocytes, accumulating in brain lesions of Creutzfeldt-Jakob disease and multiple sclerosis, suggesting a role of AQP1-expressing astrocytes in brain water homeostasis under pathological conditions. To clarify a pathological implication of AQP1 in Alzheimer disease (AD), we investigated the possible relationship between amyloid-beta (Abeta) deposition and astrocytic AQP1 expression in the motor cortex and hippocampus of 11 AD patients and 16 age-matched other neurological disease cases. In all cases, AQP1 was expressed exclusively in a subpopulation of multipolar fibrillary astrocytes. The great majority of AQP1 expressing astrocytes were located either on the top of or in close proximity to Abeta plaques in AD brains but not in non-AD cases, whereas those independent of Abeta deposition were found predominantly in non-AD brains. By Western blot, cultured human astrocytes constitutively expressed AQP1, and the levels of AQP1 protein expression were not affected by exposure to Abeta(1-42) peptide, but were elevated by hypertonic sodium chloride. By immunoprecipitation, the C-terminal fragment-beta (CTFbeta) of amyloid precursor protein interacted with the N terminal half of AQP1 spanning the transmembrane helices H1, H2 and H3. These observations suggest the possible association of astrocytic AQP1 with Abeta deposition in AD brains. PMID- 18509663 TI - Bilateral dermoid cysts of the ovary in a pregnant woman: case report and review of the literature. AB - OBJECTIVE: Most nonphysiological ovarian masses discovered during pregnancy are benign dermoid cysts. The association of dermoid cysts with pregnancy has been increasingly reported since 1918. They usually present the dilemma of weighing the risks of surgery and anesthesia versus the risks of untreated adnexal mass. METHOD: We are reporting an illustrative case and presenting a review of the literature for recommendations regarding the management of such cases. RESULT: The bilateral dermoid cysts were surgically treated in the second trimester. CONCLUSION: Most references state that it is more feasible to treat bilateral dermoid cysts of the ovaries discovered during pregnancy if they grow beyond 6 cm in diameter. This is usually performed through laparotomy or very carefully through laparoscopy and should preferably be done in the second trimester. PMID- 18509664 TI - The effect of operating time on surgeon's hand tremor. AB - The objective of this prospective study, performed at two tertiary referral centers in the West Midlands, was to determine if operating has an effect on a surgeon's baseline tremor. A total of 10 head and neck surgery consultants, 2 ENT registrars and 19 normal controls participated in the study. The interventions were preoperative and postoperative tremor measurements for surgeons and pre and post-days' desk work for controls, with the main outcomes measure being the percentage change in tremor. No difference in baseline tremor was determined between consultants and registrars. Operating led to an increase in hand tremor in all subjects. Tremor increases in all subjects were directly proportional to the length of the time spent in operating. Operating compared to a normal day's desk work increased tremor by a factor of 8.4. In conclusion, surgeons should be aware that their tremor will increase as an operation progresses. More complex parts should be performed as early in the day as possible, or, in the case of a very long operation, a change of surgeons may occasionally be necessary. PMID- 18509665 TI - Investigation continues on the atomic bomb dosimetry. PMID- 18509666 TI - Hydrogen analysis for granite using proton-proton elastic recoil coincidence spectrometry. AB - In an effort to develop DS02, a new radiation dosimetry system for the atomic bomb survivors of Hiroshima and Nagasaki, measurements of neutron-induced activities have provided valuable information to reconstruct the radiation situation at the time of the bombings. In Hiroshima, the depth profile of (152)Eu activity measured in a granite pillar of the Motoyasu Bridge (128 m from the hypocenter) was compared with that calculated using the DS02 methodology. For calculation of the (152)Eu production due to the thermal-neutron activation reaction, (151)Eu(n,gamma)(152)Eu, information on the hydrogen content in granite is important because the transport and slowing-down process of neutrons penetrating into the pillar is strongly affected by collisions with the protons of hydrogen. In this study, proton-proton elastic recoil coincidence spectrometry has been used to deduce the proton density in the Motoyasu pillar granite. Slices of granite samples were irradiated by a 20 MeV proton beam, and the energies of scattered and recoil protons were measured with a coincidence method. The water concentration in the pillar granite was evaluated to be 0.30 +/- 0.07%wt. This result is consistent with earlier data on adsorptive water (II) and bound water obtained by the Karl Fisher method. PMID- 18509667 TI - Experimental study on antiviral activity of silicone oil in vitro. AB - OBJECTIVE: To study the antiviral activity of silicone oil against herpes simplex virus-1 (HSV-1) in vitro. METHODS: (1) TCID(50) (Tissue culture infectious dose 50% endpoint) of HSV-1 was titrated. (2) The Hela cells were placed in a 96-well plate. (3) The virus was diluted by 100TCID(50), 50TCID(50), 30TCID(50), 10TCID(50), and 1TCID(50) individually. (4) Each of the five different concentrations of virus was inoculated into 16 wells. Of the 16 wells, 0.1 ml silicone oil was added to eight of them as the experimental group, and the other eight wells were used as controls. (5) Sixteen additional wells were added: silicone oil and maintenance media were added to eight wells, and only maintenance media to the other eight wells. RESULTS: (1) The cytopathic effect (CPE) of wells inoculated with 30TCID(50) combined with silicone oil was significantly less than that of the viral control at 32 hours (P < 0.01), and the same results occurred in group 10TCID(50) combined with silicone oil at 45 hours (P < 0.01) and group 1TCID(50) combined with silicone oil at 51 hours (P < 0.01). (2) In the viral control, the cells in 30TCID(50), 10TCID(50) and 1TCID(50) had pathological changes at 45, 58 and 58 hours respectively. (3) The cells of either the viral control group or group 50TCID(50) and 100TCID(50) had pathological changes at 32 hours (P > 0.05). CONCLUSION: Silicone oil has an antiviral function against HSV-1. The antiviral effect of silicone oil is correlated with concentration of virus. PMID- 18509668 TI - Foveal RPE autofluorescence as a prognostic factor for anti-VEGF therapy in exudative AMD. AB - BACKGROUND: Autofluorescence (AF) of the retinal pigment epithelium (RPE) are thought to reflect metabolic activity of the RPE cells, which in turn is largely driven by photoreceptor outer segment renewal. In exudative AMD, choroidal new vessels (CNV) may be confined to Bruch's membrane, or transgress the RPE, with consequence loss of photoreceptor cells. It has been suggested that they may be distinguished with autofluorescence imaging. The aim of our study was to analyze the prognostic value of RPE autofluorescence in relationship to the therapeutic outcome of anti-VEGF (vascular endothelial growth factor) therapy in exudative AMD. PATIENTS AND METHODS: AF images (Heidelberg Retina Angiograph) were obtained from 95 eyes (95 patients, mean age 77.64 years, 39 male and 56 female) with exudative macular lesions and associated drusen before therapy with intravitreal Bevacizumab (Avastin). Increased, normal, or decreased AF of a central area with diameters of 500 and 1,000 microm around the foveola were distinguished, and compared with the outcome of central vision. As a measure of data reproducibility (inter- and intraobserver variability), the kappa statistics (K > 0.6 "good", K > 0.8 "excellent") and exact agreement in % were calculated. RESULTS: Analysis of AF showed a significant difference in outcome of visual acuity in eyes with changes in AF of the central 500 and 1000 microm (Mann-Whitney test, p500 mum < 0.001, p1,000 microm = 0.02). Comparison of eyes with increased AF to the other eyes also resulted a significant difference in visual acuity at follow-up (p (incr) < 0.001); those with decreased AF had no significant difference to the eyes with normal or increased AF (p (decr) = 0.1733). CONCLUSIONS: The RPE-AF of exudative AMD lesions varies greatly. The AF differences probably represent different kinds of metabolism disorders in the RPE. Furthermore, they apparently have a great influence on the chances of anti- vascular endothelial growth factor (VEGF) therapy success; in particular the development of visual acuity is less favorable in eyes with initially increased central AF. PMID- 18509669 TI - Response of lipid, lipoprotein-cholesterol, and electrophoretic characteristics of lipoproteins following a single bout of aerobic exercise in women. AB - The effects of a single session of moderate intensity (65% VO(2)max) aerobic exercise expending 500 kcal of energy on serum lipid and lipoprotein-cholesterol concentrations and the electrophoretic characteristics of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles were determined in 11 sedentary, eumenorrheic, premenopausal women immediately prior to, and 24 and 48 h following exercise. Repeated measures analysis of variance revealed significant reductions in triglyceride (25.0%), HDL-cholesterol (10.9%), and HDL(3) cholesterol (11.9%) concentrations at 48 h post-exercise. Despite these changes in lipid and lipoprotein-cholesterol concentrations, no significant changes were observed in peak LDL or HDL particle sizes or in the distribution of cholesterol within the LDL and HDL subfractions. Accordingly, it appears that a single session of moderate intensity aerobic exercise expending 500 kcal (2,092 kJ) of energy promotes reductions in triglyceride, HDL-C, and HDL(3)-C concentrations without concomitantly affecting the electrophoretic characteristics of LDL and HDL particles in this sample of women. PMID- 18509670 TI - Extravascular perivenous fibrin support leads to aneurysmal degeneration and intimal hyperplasia in arterialized vein grafts in the rat. AB - BACKGROUND AND AIMS: External support of vein grafts by fibrin glue possibly prevents overdistension, vascular remodeling, and neointimal hyperplasia. Previous animal models of neointimal hyperplasia showed conflicting results. Here, long-term effects of external fibrin glue support were studied in a new rat model of jugular vein to abdominal aorta transposition. MATERIALS AND METHODS AND METHODS: In male Wistar rats (250-300 g) right jugular vein (1.0-1.5 cm) was transposed to the infrarenal aorta. Fibrin glue (0.25 ml) covered the vein before releasing the vascular clamps (n = 6). Control vein grafts were exposed directly to blood pressure. After 16 weeks vein grafts were pressure-fixed for histology. Intima thickness, luminal and intimal area were measured by planimetry and elastic fibers demonstrated by Elastica van Giesson staining. RESULTS: Intimal thickness (74.04 +/- 6.7 microm vs 1245 +/- 187 microm, control vs fibrin treatment; p < 0.001), intimal area (2517.16 +/- 355 mm(2) vs 18424 +/- 4927 mm(2), control vs fibrin treatment; p < 0.05) and luminal area (2184.75 +/- 347 mm(2) vs 7231.85 +/- 1782 mm(2), control vs fibrin treatment; p < 0.05) were significantly increased, elastic fibers in the vessel wall were diminished and the vessel wall infiltrated by mononuclear cells in fibrin glue supported veins. CONCLUSION: External support of vein grafts by fibrin glue leads to aneurysmal degeneration and intimal hyperplasia, thereby possibly jeopardizing long-term graft patency. PMID- 18509671 TI - Expression of the muscular dystrophy-associated caveolin-3(P104L) mutant in adult mouse skeletal muscle specifically alters the Ca(2+) channel function of the dihydropyridine receptor. AB - Caveolins are plasma-membrane-associated proteins potentially involved in a variety of signalling pathways. Different mutations in CAV3, the gene encoding for the muscle-specific isoform caveolin-3 (Cav-3), lead to muscle diseases, but the underlying molecular mechanisms remain largely unknown. Here, we explored the functional consequences of a Cav-3 mutation (P104L) inducing the 1C type limb girdle muscular dystrophy (LGMD 1C) in human on intracellular Ca(2+) regulation of adult skeletal muscle fibres. A YFP-tagged human Cav-3(P104L) mutant was expressed in vivo in muscle fibres from mouse. Western blot analysis revealed that expression of this mutant led to an approximately 80% drop of the level of endogenous Cav-3. The L-type Ca(2+) current density was found largely reduced in fibres expressing the Cav-3(P104L) mutant, with no change in the voltage dependence of activation and inactivation. Interestingly, the maximal density of intramembrane charge movement was unaltered in the Cav-3(P104L)-expressing fibres, suggesting no change in the total amount of functional voltage-sensing dihydropyridine receptors (DHPRs). Also, there was no obvious alteration in the properties of voltage-activated Ca(2+) transients in the Cav-3(P104L)-expressing fibres. Although the actual role of the Ca(2+) channel function of the DHPR is not clearly established in adult skeletal muscle, its specific alteration by the Cav-3(P104L) mutant suggests that it may be involved in the physiopathology of LGMD 1C. PMID- 18509672 TI - Substance P stimulates CFTR-dependent fluid secretion by mouse tracheal submucosal glands. AB - The mucosa of the proximal airways defends itself and the lower airways from inhaled irritants such as capsaicinoids, allergens, and infections by several mechanisms. Sensory nerves monitor the luminal microenvironment and release the tachykinin substance P (SP) to stimulate mucus secretion. Here, we have studied the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in SP stimulation by comparing mouse airway submucosal gland responses in wild-type (WT) and CFTR-/- mice. Capsaicinoids (chili pepper oil) increased fluid secretion by glands from WT mice five-fold, and this response was abolished by exposing the basolateral aspect of the tracheas to L-732,138 (10 micromol/l), a specific antagonist of the neurokinin-1 receptor. Secretion was also stimulated 25-fold by basolateral application of SP, and this response was strongly inhibited by the CFTR inhibitor CFTR(inh)172. In contrast, submucosal glands from CFTR knockout mice failed to secrete when stimulated by SP (1 micromol/l), although those from wild-type control littermates were responsive. SP stimulation of wild-type glands was also abolished by clotrimazole (25 micromol/l), a blocker of Ca(2+)-activated K(+) channels. These results indicate that SP mediates local responses to capsaicinoids through a mechanism involving coordinated activation of CFTR and K(+) channels. To our knowledge, this is the first study in which CFTR-dependent responses to substance P have been directly demonstrated. Since CFTR regulation is qualitatively similar in human and mouse glands, loss of this local regulation in CF may contribute to reduced innate defenses in CF airways. PMID- 18509673 TI - Granulomatous hepatitis, perihepatic lymphadenopathies, and autoantibody positivity: an unusual association in a child with hepatitis C. AB - A 10-year-old boy with hepatitis C had granulomatous hepatitis (GH) at initial liver biopsy. He also had enlarged perihepatic lymph nodes and smooth muscle antibody (SMA) positivity. GH is a rare finding in hepatitis C virus (HCV) infection. Our patient is special since GH secondary to HCV infection was associated with both autoantibodies and multiple intraabdominal lymphadenopathies. After interferon (IFN) and ribavirin therapy, HCV RNA became negative, along with the resolution of hepatic granulomas (HG), lymphadenopathies, and SMA positivity. Although early virologic response was not achieved under IFN treatment, the therapy period was extended, contrary to routine practice, and resulted in a delayed response. We conclude that the usage of IFN for longer periods in GH-associated HCV infection might be promising. PMID- 18509674 TI - Cardiac arrest following naloxone in an extremely preterm neonate. AB - Naloxone is a pure opioid antagonist specifically indicated for respiratory depression due to opioid exposure. There is not enough data on safety of naloxone, especially in extremely preterm neonates. We report the case of a preterm neonate (gestation 27 weeks and 3 days, birth weight 485 g) who developed cardiac arrest following treatment with naloxone (dose 100 mcg/kg) for a tenfold morphine overdose on day 7 while being ventilated for respiratory distress syndrome. Asystolic cardiac arrest occurred immediately after administering naloxone and required full resuscitation, including adrenaline and external chest compression. Recovery from the cardiac arrest was complete. However, the neonate died on day 45 of life due to unrelated causes. Reports of similar adverse effects of the drug in adults as well as data from animal models raise concerns about safety of naloxone, especially in preterm neonates, given the lack of data on the pharmacokinetics of the drug in this high-risk population. Possible explanations for the adverse event under such a scenario include an idiosyncratic reaction, hypoxia, direct myocardial depressant effect and sympathoadrenal interactions. Awareness of this rare but potentially lethal complication of naloxone is necessary to optimise the response to such an adverse event. PMID- 18509675 TI - Clinical heterogeneity can hamper the diagnosis of patients with ZAP70 deficiency. AB - One of the severe combined immunodeficiencies (SCIDs), which is caused by a genetic defect in the signal transduction pathways involved in T-cell activation, is the ZAP70 deficiency. Mutations in ZAP70 lead to both abnormal thymic development and defective T-cell receptor (TCR) signaling of peripheral T-cells. In contrast to the lymphopenia in most SCID patients, ZAP70-deficient patients have lymphocytosis, despite the selective absence of CD8+ T-cells. The clinical presentation is usually before 2 years of age with typical findings of SCID. Here, we present three new ZAP70-deficient patients who vary in their clinical presentation. One of the ZAP70-deficient patients presented as a classical SCID, the second patient presented as a healthy looking wheezy infant, whereas the third patient came to clinical attention for the eczematous skin lesions simulating atopic dermatitis with eosinophilia and elevated immunoglobulin E (IgE), similar to the Omenn syndrome. This study illustrates that awareness of the clinical heterogeneity of ZAP70 deficiency is of utmost importance for making a fast and accurate diagnosis, which will contribute to the improvement of the adequate treatment of this severe immunodeficiency. PMID- 18509676 TI - Necrotising enterocolitis in a term neonate with trisomy 21 exposed to maternal HIV and antiretroviral medication. AB - Necrotising enterocolitis (NEC) was diagnosed in a term infant with Down's syndrome exposed to maternal human immunodeficiency virus (HIV) infection and antiretroviral therapy. PMID- 18509677 TI - Acaricidal activity of the hyacinthacine analogues derived from pyrrolizidine alkaloids on Rhipicephalus (Boophilus) microplus. AB - This study reports the effect of six hyacinthacine analogues derived from pyrrolizidine alkaloids on egg hatchability and mortality rates of newly hatched larvae of the cattle tick Rhipicephalus (Boophilus) microplus. All the compounds were toxic to the larvae of the ticks and inhibited the eggs hatchability in the higher concentration tested (5 microg/ml). Even in the lowest concentration (0.625 microg/ml), some effect in the eggs hatchability was observed. PMID- 18509678 TI - Decreased basal non-insulin-stimulated glucose uptake by diaphragm in streptozotocin-induced diabetic mice infected with Schistosoma mansoni. AB - It had been suggested that chronic exposure to Schistosoma mansoni prevents the onset of Th1-mediated diseases such as diabetes mellitus. The present study was carried out on four groups of mice: (1) control group, (2) group infected with S. mansoni, (3) group injected with streptozotocin to induce diabetes, and (4) group infected and then 3 months postinfection injected with streptozotocin. No differences were detected between the infected non-diabetic and infected diabetic groups regarding worm burden, tissue egg count, and oogram. At the same time, results showed a reducing effect of S. mansoni infection on the rate of glucose uptake by the diaphragm with reduction in glycogen content of soleus muscle. This an important issue since skeletal muscle is the primary site for insulin stimulated glucose disposal. In conclusion, because of the detected depressed peripheral glucose uptake by the diaphragm, the protecting effect of helminths infection in diabetes should be reconsidered, to be able to devise therapeutic strategies for the treatment of autoimmune diseases. PMID- 18509679 TI - Learning and orientation to odor in the bug Rhodnius prolixus Stal 1859 under laboratory conditions. AB - Two experiments are described investigating learning and orientation in the triatomine Rhodnius prolixus. In experiment 1, Pavlovian conditioning was investigated. The experiment differed from our previous work in that the intensity of the conditioned and unconditioned stimuli were reduced and the number of training trials increased. Once again, no evidence for Pavlovian conditioning was found. In experiment 2, an "orientation arena" was developed in which the orientation of R. prolixus to a human forearm was investigated when an area of the forearm was impregnated with the odor of ruda or almizcle compared to a forearm with no odor. The various paths of the animal from the bottom of the arena until ascending and piercing the forearm, located at the top of the arena, was scored using a grid system and videotaped. The results indicated that under the no odor condition R. prolixus predominately travels in a straight line from the bottom of the arena to the top where the forearm is located. In contrast, the most variable number of paths occurred with exposure to ruda. Exposure to almizcle elicited straight line paths but other paths were evident, although not as variable as that observed with ruda. PMID- 18509680 TI - Identification of Leishmania parasites in clinical samples obtained from cutaneous leishmaniasis patients using PCR-RFLP technique in endemic region, Sanliurfa province, in Turkey. AB - Antroponotic cutaneous leishmaniasis (ACL) is an endemic disease and one of the major health problems in Sanliurfa province located in the southeastern region of Turkey. Leishmania tropica is confirmed as the causative agent of ACL in this region. In Sanliurfa city alone, the recorded total cases of ACL were 6,817 between 2001 and 2006. We aimed to determine the effectiveness of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identification and differentiation of the Leishmania parasite in comparison to direct microscopic examination of clinical samples. The lesion exudates were collected from 51 ACL suspected patients and used for smear-slide preparations and DNA isolation. The isolated DNA was amplified by PCR, including primers selected on repetitive DNA for identification of a Leishmania subgenus, and the amplified DNA was restricted by HaeIII restriction endonuclease. The PCR-RFLP results showed that only L. tropica exists in this province. It is also determined that the positivity rate with PCR was higher (96%) than by microscopic examination (64%) in the diagnosis of ACL. Our results indicate that the PCR-RFLP method is more sensitive and specific for the detection and differentiation of agents of ACL in this area. PMID- 18509681 TI - Do two mutually exclusive gene modules define the phenotypic diversity of mammalian smooth muscle? AB - Smooth muscle cells (SMCs) are key components of all hollow organs, where they perform contractile, synthetic and other functions. Unlike other muscle cells, SMCs are not terminally differentiated, but exhibit considerable phenotypic variation. Such variation is manifested both across disease states such as asthma and atherosclerosis, and physiological states such as pregnancy and wound healing. While there has been considerable investigation into the diversity of SMCs at the level of morphology and individual biomarkers, less is known about the diversity of SMCs at the level of the transcriptome. To explore this question, we performed an extensive statistical analysis that integrates 200 transcriptional profiles obtained in different SMC phenotypes and reference tissues. Our results point towards a non-trivial hypothesis: that transcriptional variation in different SMC phenotypes is characterized by coordinated differential expression of two mutually exclusive (anti-correlating) gene modules. The first of these modules (C) encodes 19 co-transcribed cell cycle associated genes, whereas the other module (E) encodes 41 co-transcribed extra cellular matrix components. We propose that the positioning of smooth muscle cells along the C/E axis constitutes an important determinant of SMC phenotypes. In conclusion, our study introduces a new approach to assess phenotypic variation in smooth muscle cells, and is relevant as an example of how integrative bioinformatics analysis can shed light on not only terminal differentiated states but also subtler details in phenotypic variability. It also raises the broader question whether coordinated expression of gene modules is a common mechanism underlying phenotypic variability in mammalian cells. PMID- 18509682 TI - The phytoestrogens daidzein and genistein enhance the insulin-stimulated sulfate uptake in articular chondrocytes. AB - Clinical observations have suggested a relationship between osteoarthritis and a changed estrogen metabolism in menopausal women. Phytoestrogens have been shown to ameliorate various menopausal symptoms. Proteoglycans (PG) consisting of low and high sulfated glycosaminoglycans (GAG) are the main components of articular cartilage matrix, and their synthesis is increased by insulin in growth plate cartilage. We have investigated whether GAG synthesis and sodium [35S]sulfate incorporation in female bovine articular chondrocytes are affected by daidzein, genistein, and/or insulin. For comparative purposes, estradiol incubations were performed. Articular chondrocytes were cultured in monolayers at 5% O2 and 5% CO2 in medium containing serum for 7 days followed by the addition of 10(-11) M-10( 4) M daidzein, genistein, 17beta-estradiol, or 5 microg/ml insulin in a serum free culture phase of 2 days. Photometrically analyzed GAG synthesis was significantly suppressed by high doses (10(-5) M-10(-4) M) of daidzein, genistein, and 17beta-estradiol. Although insulin raised the sodium [35S]sulfate uptake significantly, different concentrations of daidzein, genistein, or 17beta estradiol showed no significant effects. However, the stimulating effect of insulin on sulfate incorporation was enhanced significantly after preincubation of cells with 10(-11) M-10(-5) M daidzein or 10(-9) M-10(-5) M genistein but not by 17beta-estradiol. In view of the risks of long-term estrogen replacement therapy, further experiments should clarify the potential benefit of phytoestrogens and insulin in articular cartilage metabolism. PMID- 18509683 TI - Clinical outcomes in pediatric hemodialysis patients in the USA: lessons from CMS' ESRD CPM Project. AB - Although prospective randomized trials have provided important information and allowed the development of evidence-based guidelines in adult hemodialysis (HD) patients, with approximately 800 prevalent pediatric HD patients in the United States, such studies are difficult to perform in this population. Observational data obtained through the Center for Medicare & Medicaid Services' (CMS') End Stage Renal Disease (ESRD) Clinical Performance Measures (CPM) Project have allowed description of the clinical care provided to pediatric HD patients as well as identification of risk factors for failure to reach adult targets for clinical parameters such as hemoglobin, single-pool Kt/V (spKt/V) and serum albumin. In addition, studies linking data from the ESRD CPM Project and the United States Renal Data System have allowed evaluation of associations between achievement of those targets and the outcomes of hospitalization and death. The results of those studies, while unable to prove cause and effect, suggest that the adult ESRD CPM targets may assist in identifying pediatric HD patients at risk for poor outcomes. PMID- 18509684 TI - Lipid peroxidation and antioxidant activity in saliva of periodontitis patients: effect of smoking and periodontal treatment. AB - The aim of this study was to measure lipid peroxidation (as an end product of oxidative stress) and corresponding antioxidant activity in patients with periodontitis and assess the influence of smoking and periodontal treatment on these parameters. Thirty healthy subjects (including 15 smokers) were compared to periodontitis patients (n = 30, including 15 smokers). Malondialdehyde (MDA), glutathione peroxidase (GSHPx) and the total antioxidant capacity (TAOC) were recorded in saliva. The lowest level of lipid peroxidation (MDA) was measured in saliva in the non-smoking periodontally healthy subjects (0.065 +/- 0.05 micromol/l). MDA levels were significantly higher in periodontitis patients who smoked (0.123 +/- 0.08 micromol/l) compared to non-smoking controls (0.065 +/- 0.05 micromol/l; p < 0.05). The periodontally healthy subjects demonstrated significantly lower levels of GSHPx (antioxidative parameter) than the periodontitis group (p < 0.05). The TAOC flow rate (delivered antioxidant components within saliva) was significantly lower in patients with periodontitis (0.34 +/- 0.26 micromol/ml) in comparison to the controls (0.62 +/- 0.24 micromol/ml; p < 0.05). Patients with periodontitis demonstrate more lipid peroxidation than healthy subjects, and this effect is enhanced by smoking. Imbalance between oxidative stress and antioxidant capacity may play a role in the pathogenesis of periodontal disease. Non-surgical periodontal treatment leads to a reduction of MDA and GSHPx to levels comparable to healthy controls. PMID- 18509685 TI - Influence of amino nitrogen in the culture medium enhances the production of delta-endotoxin and biomass of Bacillus thuringiensis var. israelensis for the large-scale production of the mosquito control agent. AB - We reported here the role of amino nitrogen in the commercial production of Bacillus thuringiensis var. israelensis media design. The insect pathogen B. thuringiensis var. israelensis was cultured in different media containing varying initial levels of amino nitrogen sources obtained from three different commercial venders. The biomass, mosquito larval toxicity and spore count produced were measured during the fermentation process. The results showed that the higher level of initial amino nitrogen concentrations in the medium led to higher yield of biomass (dry weight 4.78 g l(-1)), larvicidal activity (LC(50) 18.52 ng ml( 1)) and spore count (3.24 x 10(11) CFU ml(-1)). Similarly decreasing the initial amino nitrogen concentration in the medium led to a decreased biomass (dry weight 1.64 g l(-1)), larvicidal activity (LC(50) 27.01 ng ml(-1)) and spore count (3.7 x 10(10) CFUml(-1)). PMID- 18509686 TI - A low noise remotely controllable wireless telemetry system for single-unit recording in rats navigating in a vertical maze. AB - The use of cables for recording neural activity limits the scope of behavioral tests used in conscious free-moving animals. Particularly, cable attachments make it impossible to record in three-dimensional (3D) mazes where levels are vertically stacked or in enclosed spaces. Such environments are of particular interest in investigations of hippocampal place cells, in which neural activity is correlated with spatial position in the environment. We developed a flexible miniaturized Bluetooth-based wireless data acquisition system. The wireless module included an 8-channel analogue front end, digital controller, and Bluetooth transceiver mounted on a backpack. Our bidirectional wireless design allowed all data channels to be previewed at 1 kHz sample rate, and one channel, selected by remote control, to be sampled at 10 kHz. Extracellular recordings of neuronal activity are highly susceptible to ambient electrical noise due to the high electrode impedance. Through careful design of appropriate shielding and hardware configuration to avoid ground loops, mains power and Bluetooth hopping frequency noise were reduced sufficiently to yield signal quality comparable to those recorded by wired systems. With this system we were able to obtain single unit recordings of hippocampal place cells in rats running an enclosed vertical maze, over a range of 5 m. PMID- 18509687 TI - The ApoC-I content of VLDL particles is associated with plaque size in persons with carotid atherosclerosis. AB - Previous studies have shown that postprandial triglyceride-rich lipoproteins (TRLs) are enriched with apolipoprotein-C-I (apoC-I) in healthy individuals with increased intima-media thickness and in patients with coronary artery disease. The purpose of the present study was to determine apoC-I in TRL in persons with carotid atherosclerosis and its relation to plaque area. A population-based case (n = 42)-control (n = 39) study was conducted in persons with carotid atherosclerosis, assessed by B-mode ultrasound, and healthy controls. VLDL (Sf 20 400) was isolated in the fasting state and 4 h after ingestion of a standard fat meal. In the fasting state, persons with carotid atherosclerosis had increased number of apoC-I per VLDL-particle compared to persons without carotid atherosclerosis (8.6 +/- 5.4 vs. 6.3 +/- 4.2, P = 0.018). Total plaque area increased linearly (P = 0.017) across tertiles of apoC-I per VLDL-particle. In the postprandial state, a similar increase in the number of apoC-I per VLDL particle occurred in both cases and controls (P < 0.001), but no significant difference was observed between groups. The number of apoC-I per VLDL-particle in the fasting state was accompanied by delayed clearance of TRL in the postprandial state, and associated with cholesterol enrichment of the VLDL-particles. Our findings support the concept that the number of apoC-I per VLDL-particle may be of importance for initiation and progression of atherosclerosis. PMID- 18509688 TI - Farnesol decreases serum triglycerides in rats: identification of mechanisms including up-regulation of PPARalpha and down-regulation of fatty acid synthase in hepatocytes. AB - Obesity is associated with impaired fatty acid (FA) oxidation and increased de novo hepatic lipogenesis that may contribute to the development of hypertriglyceridemia, an important risk factor for the development of cardiovascular disease. Strategies to improve hepatocyte FA metabolism, including dietary interventions, are therefore important for the prevention of obesity associated co-morbidities. Farnesol is consumed in the diet as a component of plant products. In the present study, we administered farnesol orally to rats for seven days and found significantly reduced serum triglyceride concentrations compared with controls. Potential mechanisms underlying the hypotriglyceridemic effect of farnesol were investigated using clone-9 cultured rat hepatocytes. Farnesol significantly upregulated expression of peroxisome proliferator activated receptor alpha (PPARalpha) and the PPARalpha-regulated genes fatty acyl CoA oxidase and carnitine palmitoyl transferase 1a, suggesting that increased hepatic FA oxidation may contribute to serum triglyceride lowering in rats. Farnesol did not change SREBP-1c mRNA levels, but significantly down-regulated fatty acid synthase (FAS) mRNA and protein levels and activity, indicating that attenuated lipogenesis may also contribute to hypotriglyceridemic effects of farnesol in vivo. Rescue experiments revealed that down-regulation of FAS by farnesol was not related to activation of PPARalpha, but rather was caused by a 9 cis retinoic acid mediated mechanism that involved down-regulation of retinoid X receptor beta. Diets rich in plant products are associated with a lower risk of cardiovascular disease. Our findings suggest that farnesol may contribute to this protective effect by lowering serum TG levels. PMID- 18509689 TI - Increase of serum cholesterol levels by heat-moisture-treated high-amylose cornstarch in rats fed a high-cholesterol diet. AB - The effects of four cornstarches containing various contents of resistant starch on serum and liver cholesterol levels in rats fed a high-cholesterol diet were investigated. Male Sprague Dawley rats (aged 4 weeks) were divided into four groups (n = 7) and fed high-cholesterol diets containing 15% of cornstarch (CS), heat-moisture-treated CS (HCS), high-amylose CS (HA), or heat-moisture-treated HA (HHA) for 21 days. The results showed that the serum and hepatic level of total cholesterol, LDL-cholesterol, and triglyceride in rats of the HHA group and their arteriosclerosis index were significantly higher, suggesting that HHA increases the risk of arteriosclerosis under a high-cholesterol dietary condition. No significant between-group differences were noted in the levels of plasma mevalonic acid and hepatic HMG-CoA reductase mRNA, whereas fecal cholesterol excretion was significantly higher in the HHA group, indicating that the elevation of the serum and liver cholesterol levels was not due to the promotion of liver cholesterol synthesis and cholesterol absorption in the intestine. PMID- 18509690 TI - Vitamin E transfer from lipid emulsions to plasma lipoproteins: mediation by multiple mechanisms. AB - The present study determined alpha-tocopherol mass transfer from an alpha tocopherol-rich emulsion to LDL and HDL, and assessed the potential of different mechanisms to modulate alpha-tocopherol transfers. Emulsion particles rich in alpha-tocopherol were incubated in vitro with physiological concentrations of LDL or HDL. The influence of plasma proteins was assessed by adding human lipoprotein poor plasma (LPP) fraction with intact vs heat inactivated PLTP, or with a specific cholesteryl ester transfer protein (CETP) inhibitor, or by adding purified PLTP or pig LPP which lacks CETP activity. After 4 h incubation in absence of LPP, alpha-tocopherol content was increased by ~80% in LDL and ~160% in HDL. Addition of LPP markedly enhanced alpha-tocopherol transfer leading to 350-400% enrichment in LDL or HDL at 4 h. Higher (~10 fold) enrichment was achieved after 20 h incubation with LPP. Facilitation of alpha-tocopherol transfer was (i) more than 50% higher with human vs pig LPP (despite similar PLTP phospholipid transfer activity), (ii) reduced by specific CETP activity inhibition, (iii) not fully suppressed by heat inactivation, and (iv) not restored by purified PLTP. In conclusion, alpha-tocopherol content in LDL and HDL can be markedly raised by rapid transfer from an alpha-tocopherol-rich emulsion. Our results indicate that alpha-tocopherol mass transfer between emulsion particles and lipoproteins is mediated by more than one single mechanism and that this transfer may be facilitated not only by PLTP but likely also by other plasma proteins such as CETP. PMID- 18509691 TI - Compression of the deep branch of the ulnar nerve in Guyon's canal by a ganglion: two cases. AB - INTRODUCTION: Ulnar nerve compression at the wrist can be caused by a variety of intrinsic and extrinsic factors. Isolated compression of only the deep branch of ulnar nerve by a ganglion is very uncommon. Ultrasound examination can clearly show the cystic lesion compressing the nerves. MATERIALS AND METHODS: We present two cases of compression of deep branch of ulnar nerve by a ganglion in the Guyon's canal. Two male patients presented with history of progressive weakness and paraesthesia in the medial 1(1/2) digits of the non-dominant hand. Interestingly, both the patients noticed sudden onset and rapid progress of the symptoms and signs. Clinical examination revealed typical symptoms of ulnar nerve (deep branch) palsy. Nerve conduction studies showed severe denervation of the deep branch of the ulnar nerves in both the patients and ultrasound confirmed the diagnosis. Surgical decompression led to complete recovery. RESULTS AND DISCUSSION: Whilst compression by a ganglion in the Guyon's canal is rare but well recognized, a feature of both of our cases was the rapid progression and severe nature of the compressive symptoms and signs. This is in contrast to the more typical features of compressive neuropathy and should alert the clinician to the possible underlying cause of compression. Early decompression has the potential to promote a complete recovery. PMID- 18509692 TI - Childhood nonunion of ulna presenting with wrist deformity in an adult: a case report. AB - We describe an adult patient with traumatic, nonunion of ulna sustained at 11 years of age who presented with wrist deformity. The possible pathogenesis, differential diagnoses and its successful management are described. A 23-year- old right hand dominant male presented with a progressive wrist deformity of his right upper limb. At 11 years of age, he sustained an isolated open fracture of the right forearm. He had nonoperative treatment. He had 60 degrees of ulnar deviation at wrist. He had no pain in the wrist or elbow. He was able to do all activities using his right upper limb. Radiograph revealed a nonunion of ulna in mid-shaft. The radius was bowed. Radiographs at the time of injury revealed a displaced both bones forearm fracture in mid-shaft. He underwent open reduction, internal fixation of ulna with bone grafting and a corrective osteotomy of the radius. The contracted Extensor carpi ulnaris was Z lengthened. Seven months postoperative, both the nonunion of ulna and radius osteotomy were consolidated. The wrist had no deformity. He had returned to preoperative activity level. Though nonunion is rare in pediatric forearm fractures, asymmetric bone and soft tissue growth can lead to deformities even in the absence of physeal injury. In addition to the standard treatment of nonunion, maintenance of the relative lengths of radius and ulna is essential, to obtain optimum function. PMID- 18509697 TI - Mechanical stimulation of bone formation is normal in the SAMP6 mouse. AB - With aging, the skeleton may have diminished responsiveness to mechanical stimulation. The senescence-accelerated mouse SAMP6 has many features of senile osteoporosis and is thus a useful model to examine how the osteoporotic skeleton responds to mechanical loading. We performed in vivo tibial bending on 4-month old SAMP6 (osteoporotic) and SAMR1 (control) mice. Loading was applied daily (60 cycles/day, 5 days/week) for 2 weeks at peak force levels that produced estimated endocortical strains of 1,000 and 2,000 microepsilon In a separate group of mice, sham bending was applied. Comparisons were made between right (loaded) and left (nonloaded) tibiae. Tibial bone marrow cells were cultured under osteogenic conditions and stained for alkaline phosphatase (ALP) and alizarin red (ALIZ) at 14 and 28 days, respectively. Tibiae were then embedded in plastic and sectioned, and endocortical bone formation was assessed based on calcein labels. Tibial bending did not alter the osteogenic potential of the marrow as there were no significant differences in ALP or ALIZ staining between loaded and nonloaded bones. Tibial bending activated the formation of endocortical bone in both SAMP6 and SAMR1 mice, whereas sham bending did not elicit an endocortical response. Both groups of mice exhibited bending strain-dependent increases in bone formation rate. We found little evidence of diminished responsiveness to loading in the SAMP6 skeleton. In conclusion, the ability of the SAMP6 mouse to respond normally to an anabolic mechanical stimulus distinguishes it from chronologically aged animals. This finding highlights a limitation of the SAMP6 mouse as a model of senile osteoporosis. PMID- 18509698 TI - Dried plum polyphenols inhibit osteoclastogenesis by downregulating NFATc1 and inflammatory mediators. AB - Dried plums and their polyphenols have been shown to suppress bone resorption by downregulating receptor activator NF-kappaB ligand (RANKL). Due to the anti inflammatory and antioxidant properties of these compounds, this study was designed to investigate whether dried plum polyphenols exert additional, more direct effects on osteoclasts and their precursors. RAW 264.7 macrophages were used as a model to study osteoclast precursors and osteoclast differentiation and activity. Under inflammatory conditions induced by lipopolysaccharide (LPS), polyphenols extracted from dried plum (10, 20, and 30 microg/mL) downregulated osteoclast precursor cyclooxygenase expression and nitric oxide (NO) by inhibiting inducible NO synthase. NO and tumor necrosis factor (TNF)-alpha were also suppressed in the presence of RANKL during osteoclastogenesis by the polyphenols. Increased TNF-alpha production in response to oxidative stress, but not LPS, was decreased over time. As expected, LPS and H2O2 significantly increased the number of tartrate-resistant acid phosphatase-positive cells by 127% and 30%, respectively. Dried plum polyphenols decreased osteoclast differentiation under normal as well as inflammatory and oxidative stress conditions, coincident with the suppression of the transcription factor, nuclear factor for activated T cells (NFATcl). These inhibitory effects on osteoclastogenesis were confirmed in primary bone marrow cultures. Resorption pit formation was decreased to a similar extent as osteoclast differentiation, suggesting that dried plum polyphenols primarily affect osteoclast differentiation as opposed to activity. Our data demonstrate that dried plum polyphenols directly inhibit osteoclastogenesis, leading to a decrease in osteoclast activity, by downregulating NFATc1 and inflammatory mediators. PMID- 18509699 TI - Fat embolism syndromes following liposuction. AB - Fat embolism syndrome (FES) after liposuction is likely a life-threatening disorder, though its incidence is low. The three chief clinical manifestations include respiratory insufficiency, cerebral involvement, and petechial rash. Although FES is a multisystem disorder, the most seriously affected organs are the lungs, brain, cardiavascular system, and skin. Many laboratory findings are characteristic but nonspecific. The pathogenesis of FES after liposuction has been looked at both mechanically and biochemically. Diagnosis is difficult; Gurd and Wilson's diagnostic criteria based on clinical examination is still extensively used in clinics at present. There is no specific therapy for FES after liposuction for the moment, so prevention, early diagnosis, and supportive therapies are important. In this article we discuss the clinical presentation, pathogensis, and current methods to prevent FES and, if possible, ways to treat this complication. PMID- 18509700 TI - Landsat ETM+ images in the estimation of seasonal lake water quality in boreal river basins. AB - We investigated the use of Landsat ETM+ images in the monitoring of turbidity, colored dissolved organic matter (CDOM), and Secchi disk transparency (Z(SD)) in lakes of two river basins located in southern Finland. The ETM+ images were acquired in May, June, and September 2002 and were corrected for atmospheric disturbance using the simplified method of atmospheric correction (SMAC) model. The in situ measurements consisted of water sampling in the largest lake of the region, routine monitoring results for the whole study area, and Z(SD) observations made by volunteers. The ranges of the water quality variables in the dataset were as follows: turbidity, 0.6-25 FNU; absorption coefficient of CDOM at 400 nm, 1.0-12.2 m(-1); Z(SD), 0.5-5.5 m; and chlorophyll a concentration, 2.4-80 microg L(-1). The estimation accuracies of the image-specific empirical algorithms expressed as relative errors were 23.0% for turbidity, 17.4% for CDOM, and 21.1% for Z(SD). If concurrent in situ measurements had not been used for algorithm training, the average error would have been about 37%. The atmospheric correction improved the estimation accuracy only slightly compared with the use of top-of-atmospheric reflectances. The accuracy of the water quality estimates without concurrent in situ measurements could have been improved if in-image atmospheric parameters had been available. The underwater reflectance simulations of the ETM+ channel wavelengths using water quality typical for Finnish lakes (data from 1113 lakes) indicated that region-specific algorithms may be needed in other parts of the country, particularly in the case of Z(SD). Despite the limitations in the spectral and radiometric resolutions, ETM+ imagery can be an effective aid, particularly in the monitoring and management of small lakes (<1 km(2)), which are often not included in routine monitoring programs. PMID- 18509701 TI - Primary infrainguinal subintimal angioplasty in diabetic patients. AB - The aim of this study was to prospectively evaluate technical and clinical results of infrainguinal subintimal angioplasty in a series of diabetic patients with limb-threatening ischemia. From July 2003 to December 2007, 60 consecutive diabetic patients (M/F = 41/19; mean age, 69.4 +/- 9.4 years) with Fontaine stage IV critical limb ischemia, not suitable for surgical recanalization, underwent primary infrainguinal subintimal angioplasty. The technical success, perioperative morbidity and mortality, and clinical success (defined by ulcer healing) were evaluated. Kaplan-Meier life-table analysis was obtained for cumulative clinical success, limb salvage, and survival rates. The procedure was technically successful in 55 of 60 (91.7%) patients; in 5 cases we were not able to achieve a reentry. Periprocedural mortality was 5% (3 patients); three patients (5%) required major amputation periprocedurally. Mean follow-up was 23 months (range, 0-48 months). On an intention-to-treat basis, the limb salvage rate was 93.3% (56/60 patients); ulcer healing was observed in 45 of 60 (75%) patients and it was significantly (p < 0.05) associated with serum creatinine and HbA1c levels, diabetes duration, and infrapopliteal recanalization. One- and three-year cumulative survival rates were 91.5% and 83.1%, respectively; serum creatinine levels, patient age, and clinical success were significant predictors of survival. In conclusion, infrainguinal primary subintimal angioplasty is a safe and effective treatment in diabetic patients with limb-threatening ischemia not suitable for surgical recanalization. This procedure is aimed to create a "temporary bypass" that facilitates ulcer healing. PMID- 18509702 TI - Fibromuscular dysplasia presenting with bilateral renal infarction. AB - Fibromuscular dysplasia (FMD) describes a group of conditions which cause nonatheromatous arterial stenoses, most commonly of the renal and carotid arteries, typically in young women. We report a rare case of bilateral segmental renal infarction secondary to FMD in a young male patient. His initial presentation with loin pain and pyrexia resulted in a delay in the definitive diagnosis of FMD. He was successfully treated with bilateral balloon angioplasty. The delayed diagnosis in this patient until the condition had progressed to bilateral renal infarcts highlights the need for prompt investigation and diagnosis of suspected cases of FMD. PMID- 18509703 TI - Traversing tight ureteric strictures. PMID- 18509704 TI - Radiographic response to yttrium-90 radioembolization in anterior versus posterior liver segments. AB - The purpose of our study was to determine if preferential radiographic tumor response occurs in tumors located in posterior versus anterior liver segments following radioembolization with yttrium-90 glass microspheres. One hundred thirty-seven patients with chemorefractory liver metastases of various primaries were treated with yttrium-90 glass microspheres. Of these, a subset analysis was performed on 89 patients who underwent 101 whole-right-lobe infusions to liver segments V, VI, VII, and VIII. Pre- and posttreatment imaging included either triphasic contrast material-enhanced CT or gadolinium-enhanced MRI. Responses to treatment were compared in anterior versus posterior right lobe lesions using both RECIST and WHO criteria. Statistical comparative studies were conducted in 42 patients with both anterior and posterior segment lesions using the paired sample t-test. Pearson correlation was used to determine the relationship between pretreatment tumor size and posttreatment tumor response. Median administered activity, delivered radiation dose, and treatment volume were 2.3 GBq, 118.2 Gy, and 1,072 cm(3), respectively. Differences between the pretreatment tumor size of anterior and posterior liver segments were not statistically significant (p = 0.7981). Differences in tumor response between anterior and posterior liver segments were not statistically significant using WHO criteria (p = 0.8557). A statistically significant correlation did not exist between pretreatment tumor size and posttreatment tumor response (r = 0.0554, p = 0.4434). On imaging follow up using WHO criteria, for anterior and posterior regions of the liver, (1) response rates were 50% (PR = 50%) and 45% (CR = 9%, PR = 36%), and (2) mean changes in tumor size were -41% and -40%. In conclusion, this study did not find evidence of preferential radiographic tumor response in posterior versus anterior liver segments treated with yttrium-90 glass microspheres. PMID- 18509705 TI - Multigenic factors associated with a hydrocephalus-like phenotype found in inter subspecific consomic mouse strains. AB - Hydrocephalus is a significant clinical condition in humans and is known to be a multifactorial neurologic disorder. It has been thought that genetic factors are closely involved in the etiology of congenital hydrocephalus, but further investigation is required to elucidate the genetic architecture of hydrocephalus. By analyzing breeding records of a panel of inter-subspecific consomic mouse strains, we found that consomic strains with MSM/Ms (MSM) chromosomes 4, 5, 7, 11, 15, and 17 showed a significantly higher incidence of hydrocephalus, whereas both parental strains, MSM and C57BL/6J (B6), rarely showed this abnormality. Further analysis of the consomic Chr 17 strain revealed that apparently normal individuals of this strain also exhibited increased brain ventricle size compared to B6 and had larger individual variation of ventricle size within the strain. Thus, we concluded that hydrocephalus is an extreme phenotype of individual ventricle size variation. We then established and analyzed several subconsomic strains of Chr 17 to identify genetic factors related to hydrocephalus-like phenotype and successfully mapped one genetic locus around the proximal region of Chr 17. PMID- 18509706 TI - Outcomes following isolated limb infusion for melanoma. A 14-year experience. AB - BACKGROUND: Isolated limb infusion (ILI) is a minimally invasive technique for delivering regional chemotherapy in patients with advanced and metastatic melanoma confined to a limb. It is essentially a low-flow isolated limb perfusion (ILP) performed via percutaneous catheters without oxygenation. METHODS: From our prospective database 185 patients with advanced metastatic melanoma of the limb treated with a single ILI between 1993 and 2007 were identified. In all patients a cytotoxic drug combination of melphalan and actinomycin-D was used. Drug circulation time was 20-30 min under mild hyperthermic conditions (38-39 degrees C). RESULTS: The majority of patients (62%) were female. Their average age was 74 years (range 29-93 years). Most patients had MD Anderson stage III disease (134/185). The overall response rate was 84% [complete response (CR) rate 38%, partial response rate 46%]. Median response duration was 13 months (22 months for patients with CR; P = 0.01). Median follow-up was 20 months and median survival was 38 months. In those patients with a CR, the median survival was 53 months (P = 0.005). CR rate and survival time decreased with increasing stage of disease. On multivariate analysis significant factors for a favorable outcome were achievement of CR, stage of disease, thickness of primary melanoma, the CO(2 )level in the isolated circuit, and a Wieberdink limb toxicity score of III (considerable erythema and edema). CONCLUSION: The response rates and duration of response after ILI are comparable to those achieved by conventional ILP. ILI is a minimally invasive alternative to the much more complex and morbid conventional ILP technique for patients with advanced metastatic melanoma confined to a limb. PMID- 18509707 TI - Limits of day-before lymphoscintigraphy to localize sentinel nodes in women with cervical cancer. AB - BACKGROUND: Lymph node status in cervical cancer is a major prognostic factor. Sentinel lymph node (SN) biopsy using radiocolloid and blue dye labeling and preoperative lymphoscintigraphy has emerged as a potential alternative to systematic lymphadenectomy. The aim of this study was to evaluate the contribution of preoperative lymphoscintigraphy to SN biopsy. METHODS: Between April 2001 and December 2005, 71 of 77 patients with cervical cancer (38 patients with stages IA or IB1, and 39 patients with stage IB2, IIA or IIB) underwent laparoscopic SN procedure using radiocolloid and blue dye with day-before lymphoscintigraphy. The SN identification rates and false-negative rates were studied. RESULTS: Seventy patients underwent a combined technique and the last patient a radiocolloid technique alone due to blue dye allergic reaction. Detection rate of lymphoscintigraphy was 84.5% (60/71), with 1.4 sentinel nodes per patient. Three of 11 patients (27.3%) with no SN on lymphoscintigraphy had at least one SN during surgery. Sixteen of 27 patients (59.3%) with solitary SN on lymphoscintigraphy had multiple SNs. Nine of 35 patients (25.7%) with unilateral SNs on lymphoscintigraphy had bilateral SNs at surgery (kappa = 0.44 [0.19 0.64]). When categorized into <2 and >or=2 sentinel nodes, the correlation between lymphoscintigraphic and surgical detection was poor (kappa = 0.05 [0.0 0.18]). CONCLUSIONS: SN biopsy is a feasible and accurate method to stage early cervical cancer. However, day-before lymphoscintigraphy is poorly correlated to surgical SN mapping. PMID- 18509708 TI - Comparison of two methods of selective hepatic vascular exclusion for liver resection involving the roots of the hepatic veins. AB - BACKGROUND: Selective hepatic vascular exclusion (SHVE) is an effective hepatic vascular exclusion in controlling both inflow and outflow without interruption of caval flow, as it combines Pringle maneuver with extrahepatic selective occlusion of hepatic veins. But SHVE has not been widely used due to difficulty in extrahepatic dissection of hepatic veins. When the tumor is very close to the roots of the hepatic veins, dissecting the posterior wall of the hepatic vein may lead to rupture and massive bleeding of the hepatic vein. With our experience, clamping hepatic veins with Satinsky clamps is a safer and easier occlusion method by which the posterior wall of the hepatic veins does not need to be separated and encircled. In this report, we compared the results of selective hepatic vascular occlusion with tourniquet and Satinsky clamp for major liver resection involving the roots of the hepatic veins. METHODS: Between January 2003 to June 2006, 180 patients who underwent major liver resection with SHVE were divided into two groups according to different methods of hepatic vascular occlusion: occlusion with tourniquet (tourniquet group, n = 95) and occlusion with Satinsky clamp (Satinsky clamp group, n = 85). In the tourniquet group, the hepatic veins were encircled and occluded with tourniquet. In the Satinsky clamp group, the hepatic veins were not encircled and clamped directly by Satinsky clamp. RESULTS: Intraoperative and postoperative consequences of the patients were analyzed. The dissecting time for each hepatic vein was significantly shorter in the Satinsky group (6.2 +/- 2.4 min vs 18.3 +/- 6.2 min) than in the tourniquet group. In the tourniquet group, five hepatic veins (one right hepatic vein and four common trunk of left-middle hepatic veins) could not be dissected and encircled because the tumors involved the cava hepatic junction, and another common trunk of the left-middle hepatic vein had a small rupture during the dissection. These six patients then received successful occlusion with Satinsky clamp. There was no difference between the two groups regarding the operation duration, ischemia time, intraoperative blood loss, and postoperative complication rate. CONCLUSION: Both methods of the hepatic vein occlusion have the same effect on controlling hepatic vein bleeding, but occlusion with Satinsky clamp is safer, easier, and consumes less time in dissecting. PMID- 18509709 TI - A perspective: engineering periosteum for structural bone graft healing. AB - Autograft is superior to both allograft and synthetic bone graft in repair of large structural bone defect largely due to the presence of multipotent mesenchymal stem cells in periosteum. Recent studies have provided further evidence that activation, expansion and differentiation of the donor periosteal progenitor cells are essential for the initiation of osteogenesis and angiogenesis of donor bone graft healing. The formation of donor cell-derived periosteal callus enables efficient host-dependent graft repair and remodeling at the later stage of healing. Removal of periosteum from bone autograft markedly impairs healing whereas engraftment of multipotent mesenchymal stem cells on bone allograft improves healing and graft incorporation. These studies provide rationale for fabrication of a biomimetic periosteum substitute that could fit bone of any size and shape for enhanced allograft healing and repair. The success of such an approach will depend on further understanding of the molecular signals that control inflammation, cellular recruitment as well as mesenchymal stem cell differentiation and expansion during the early phase of the repair process. It will also depend on multidisciplinary collaborations between biologists, material scientists and bioengineers to address issues of material selection and modification, biological and biomechanical parameters for functional evaluation of bone allograft healing. PMID- 18509710 TI - Case report: meralgia paresthetica in a baseball pitcher. AB - We report a case of meralgia paresthetica occurring in an amateur baseball pitcher who experienced inguinal pain and dysesthesia in the anterolateral thigh during pitching practice. The lateral femoral cutaneous nerve was pushed up by the iliac muscle to the inguinal ligament at the sharp ridge of its fascia and ensheathed in the tendinous origin of the sartorius muscle. Neurolysis of the lateral femoral cutaneous nerve and partial dissection of the inguinal ligament and sartorius muscle promptly relieved the symptoms and the patient resumed pitching 1 month later. These anatomic variations of the lateral femoral cutaneous nerve in the inguinal region might render the nerve susceptible to compression and irritation, and repetitive contraction of inguinal muscles during throwing motion might induce and exacerbate the neuropathy of the lateral femoral cutaneous nerve. PMID- 18509711 TI - Controlled release of growth factors on allograft bone in vitro. AB - Allografts are important alternatives to autografts for treating defects after major bone loss. Bone growth factors have both local autocrine and paracrine effects and regulate the growth, proliferation, and differentiation of osteoprogenitor cells. To study the effects of prolonged, continuous, local delivery of growth factors on bone growth, we developed a new microelectromechanical system (MEMS) drug delivery device. Bone marrow cells from mice were seeded on mouse allograft discs and cultured in osteogenic media with osteogenic protein 1 (OP-1) and/or basic fibroblast growth factor (FGF-2) delivered from MEMS devices for 6 weeks. We monitored bone formation by changes of bone volume using micro-CT scanning and release of osteocalcin using ELISA. The data suggest the MEMS devices delivered constant concentrations of OP-1 and FGF-2 to the media. Bone marrow cells grew on the allografts and increased bone volume. Addition of OP-1 increased bone formation whereas FGF-2 decreased bone formation. Local delivery of growth factors over a prolonged period modulated the differentiation of osteoprogenitor cells on allograft bone. PMID- 18509712 TI - Accuracy of model-based RSA contour reduction in a typical clinical application. AB - Marker-based roentgen stereophotogrammetric analysis (RSA) is an accurate method for measuring in vivo implant migration, which requires attachment of tantalum markers to the implant. Model-based RSA allows migration measurement without implant markers; digital pose estimation, which can be thought of as casting a shadow of a surface model of the implant into the stereoradiographs, is used instead. The number of surface models required in a given clinical study depends on the number of implanted sizes and design variations of prostheses. Contour selection can be used to limit pose estimation to areas of the prosthesis that do not vary with design, reducing the number of surface models required. The effect of contour reduction on the accuracy of the model-based method was investigated using three different contour selection schemes on tibial components in 24 patients at 3 and 6 month followup. The agreement interval (mean +/- 2 standard deviations), which bounds the differences between the marker-based and model based methods with contour reduction was smaller than -0.028 +/- 0.254 mm. The data suggest that contour reduction does not result in unacceptable loss of model based RSA accuracy, and that the model-based method can be used interchangeably with the marker-based method for measuring tibial component migration. PMID- 18509714 TI - Occurrence of organ-specific and systemic autoimmune diseases among the first- and second-degree relatives of Caucasian patients with connective tissue diseases: report of data obtained through direct patient interviews. AB - Studies have demonstrated a familial aggregation of systemic and organ-specific autoimmune diseases. The aim of the present survey was to obtain, by patient interviews, a preliminary estimate of the prevalence of systemic and organ specific autoimmune diseases among the first- and second-degree relatives of Caucasian patients with connective tissue diseases (CTD) or inflammatory arthritis followed at our unit. Between June 2007 and January 2008, 626 patients and 85 controls (patients with osteoarthritis, osteoporosis, or fibromyalgia) were interviewed. Three hundred ten patients (50%) versus 21 controls (25%) were found to have at least one relative affected with an autoimmune condition (p < 0.0001). The most common conditions were organ-specific autoimmune diseases: 160 (34%) autoimmune thyroid (AT) disease, 112 (24%) psoriasis, 21 vitiligo, and 19 insulin-dependent diabetes mellitus. Systemic autoimmune diseases were reported in 126 relatives: rheumatoid arthritis (66 cases, 14%), 16 sacroileitis, and CTD (43 cases). A significant difference was observed in the prevalence of AT disease between the relatives of the patients and controls (3% versus 0.5%). In conclusion, these data confirm the high prevalence of autoimmune conditions, particularly of AT disease, among the relatives of patients. PMID- 18509713 TI - Atherosclerosis and osteoporosis: age-dependent degenerative processes or related entities? AB - Osteoporosis and atherosclerosis, two multifactorial and degenerative entities, are major public health problems. These diseases accompany the aging process and share common risk factors. Furthermore, several common pathophysiological factors have been suggested. These include similar molecular pathways involving bone and vascular mineralization, estrogen deficiency, parathyroid hormone, homocysteine, lipid oxidation products, inflammatory process, as well as vitamin D and K. Moreover, the use of statins, biphosphonates, beta-blockers and experimental dual purpose therapies based on the biological linkage of the above entities may simultaneously benefit bone loss and vascular disease. This review considers a potential link between osteoporosis and atherosclerosis beyond aging. These common factors may lead to appropriate treatment strategies. PMID- 18509715 TI - Formation of numerous rice bodies: an unusual finding of adult-onset Still's disease. AB - Adult-onset Still's disease (AOSD) is an inflammatory disorder characterized by high-spiking fever, arthritis, and an evanescent rash. The pattern of chronic arthritis is a universal feature of AOSD. However, chronic bursitis with the formation of numerous rice bodies is a very rare presentation in a patient with AOSD. To our knowledge, no case of formation of numerous rice bodies in AOSD has been reported thus far. We describe the case of a 28-year-old man with AOSD with rice bodies in his left shoulder joint. PMID- 18509716 TI - Sub-optimal pain control in patients with rheumatic disease. AB - The visual analog scale (VAS) of pain is a ubiquitous clinical and research tool with widespread application in the rheumatic diseases. The objectives of this study were to assess if patients report pain differently to doctors or nurses, to determine reproducibility of this test for diagnosis, age, gender, and treatment, and to ascertain the level of pain in patients attending general rheumatology clinics. Using a standardized line of exactly 100 mm and instructions with identical wording, consecutive patients attending general rheumatology clinics were asked to score their perceived level of pain in the preceding week. Two assessments were carried out, one before and one after the clinic visit, and each patient was questioned by both a doctor and a nurse. Differences between the first and second VAS scores (VAS1 and VAS2) were recorded. One hundred and eight patients completed the study (69 female). VAS1 and VAS2 scores were administered by a similar number of doctors and nurses. There was no significant difference between mean VAS1 and VAS2 scores (41.1 vs. 41.4 mm, p = 0.78). VAS1 and VAS2 differed by <4 mm in 59% of patients. Age, gender, or diagnosis did not influence VAS1 or VAS2. Differences in scores were independent of which health professional administered the scale (p = 0.19). Patients taking painkillers had higher mean VAS scores (49 mm) compared with those not on analgesia (27 mm; p < 0.001). Anti rheumatic treatment did not influence pain scores (p = 0.13). The VAS is a reliable and effective method of pain assessment. Results are independent of which health professional administers the scale. Patients with rheumatic disease report their pain similarly regardless of diagnosis. However, pain control is sub optimal in patients taking analgesia. Specific assessment of pain is, thus, important in patients attending rheumatology clinics. PMID- 18509717 TI - Selective induction hyperthermia following transcatheter arterial embolization with a mixture of nano-sized magnetic particles (ferucarbotran) and embolic materials: feasibility study in rabbits. AB - PURPOSE: To evaluate the possibility of selective hyperthermia following transcatheter arterial embolization (TAE) with ferucarbotran using a newly developed inductive heating (IH) device. MATERIALS AND METHODS: Twelve Japanese white rabbits were separated into four groups: those treated with TAE using a mixture of ferucarbotran and lipiodol (F-L group); those treated with ferucarbotran and gelatin sponge powder; those treated with saline and lipiodol; and a control group. These four groups received IH. Nine rabbits with renal VX2 carcinoma were separated into three groups: IH after TAE (IH-TAE tumor), TAE without IH (TAE tumor), and no treatment (control tumor). The temperature of the tumor was kept at 45 degrees C for 20 min. The therapeutic effect was pathologically evaluated by TUNEL staining. RESULTS: In the heating rates of the kidney, the F-L group showed significantly greater values than the group in which iron was not used. In the IH-TAE tumor group, tumors could be selectively heated. In TUNEL staining, the IH-TAE tumor and TAE tumor groups showed significantly greater values of apoptosis rate than in the control tumor group. CONCLUSION: IH following TAE with a mixture of ferucarbotran and lipiodol was capable of inducing selective hyperthermia with our device. However, further investigation is needed to confirm its safety and effectiveness in the treatment of malignant neoplasms in humans. PMID- 18509718 TI - Arterial catheterization and embolization for management of emergent or anticipated massive obstetrical hemorrhage. AB - PURPOSE: The aim of this study was to evaluate the indications, efficacy, and safety of arterial catheterization and embolization for the management of emergent or anticipated massive obstetrical hemorrhage and its effects on menses and fertility. MATERIALS AND METHODS: Eighteen patients underwent arterial catheterization and embolization for intractable obstetrical hemorrhage between January 2001 and December 2005. Three groups of patients were identified: group 1 (n = 6) experienced postpartum hemorrhage; group 2 (n = 5) had anticipated severe postpartum hemorrhage; and group 3 (n = 7) had a risk factor for anticipated severe hemorrhage after dilation and evacuation. Gynecological information after embolization was obtained from medical records and telephone interviews. RESULTS: All patients in group 1 had a favorable outcome after treatment with a single embolization. All patients in group 2 had a placenta previa with an estimated blood loss of 1215-3250 ml. In group 3, bleeding was controlled in six patients; one patient had a hysterectomy because embolization was not possible. There were no short-or long - term complications, and normal menstruation resumed. Four patients became pregnant after embolization. CONCLUSION: Arterial catheterization and embolization is an effective, safe method for treating intractable obstetrical hemorrhage and might eliminate the need for hysterectomy and maintain reproductive ability. PMID- 18509719 TI - Magnetic resonance imaging-guided focused ultrasound ablation of uterine fibroids: early clinical experience. AB - PURPOSE: The aim of this study was to assess the feasibility and effectiveness of magnetic resonance (MRI)-guided focused ultrasound (MRIgFUS) ablation for uterine fibroids and to identify the candidates for this treatment. MATERIALS AND METHODS: A total of 48 patients with a symptomatic uterine fibroid underwent MRIgFUS. The percent ablation volume was calculated, and the patients' characteristics and the MR imaging features of the fibroids that might predict the effect of this treatment were assessed. Changes in the symptoms related to the uterine fibroid were assessed at 6 and 12 months. RESULTS: The planned target zone were successfully treated in 32 patients with bulk-related and menstrual symptoms but unsuccessfully treated in the remaining 16 patients. These 16 patients were obese or their uterine fibroid showed heterogeneous high signal intensity on T2-weighted images. The 32 successfully treated patients were followed up for 6 months. At the 6-month follow-up, bulk-related and menstrual symptoms were diminished in 60% and 51% of patients, respectively. Among them, 17 patients were followed up for 12 months, and 9 of them who showed alleviation of bulk-related symptoms at 6 months had further improvement. The mean percent ablation volume of those nine patients was 51%. In 5 (33%) of the 15 patients with alleviation of menstrual symptoms at 6 months, the symptoms became worse at 12 months. There was a significant difference in the mean percent ablation volume between patients with alleviation of menstrual symptoms and those without (54% vs. 37%; P = 0.03). CONCLUSION: MRIgFUS ablation is a safe, effective treatment for nonobese patients with symptomatic fibroids that show low signal intensity on T2-weighted images. Ablation of more than 50% of the fibroid volume may be needed with a short-term follow-up. PMID- 18509720 TI - Angiographic evaluation of hepatic arterial damage after transarterial chemoembolization for hepatocellular carcinoma. AB - PURPOSE: The aim of this study was to assess the incidence, degree, and predictors of hepatic arterial damage (HAD) after transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: A total of 33 patients with unresectable HCC underwent TACE alone using a mixture of iodized oil, epirubicin, and gelatin sponge. A follow-up angiogram was available for 76 of 109 sessions, and HAD was evaluated at each subsegment of the hepatic artery using a three-grade scale (1, no or slight wall irregularity; 2, overt stenosis; 3, occlusion). Grades 2 and 3 were considered to indicate significant HAD. The predictors of HAD were analyzed by multivariate analysis. RESULTS: A total of 161 hepatic arteries were embolized from the lobar (n = 43), segmental (n = 40), subsegmental (n = 72), or more distal (n = 6) level. The follow-up period between the initial and last sessions ranged from 70 to 1505 days (median 497 days). Significant HAD occurred in 37 of 231 subsegmental hepatic arteries (16%) and in 16 of 33 patients (48%). The accumulated dose of epirubicin per artery (P = 0.001) and Child-Pugh score (P < 0.001) were significant predictors. CONCLUSION: TACE is more likely to induce HAD in cirrhotic patients with impaired liver function and when a high dose of the chemotherapeutic agent was used. PMID- 18509721 TI - Functional computed tomography imaging of tumor-induced angiogenesis: preliminary results of new tracer kinetic modeling using a computer discretization approach. AB - PURPOSE: The aim of this study was to establish functional computed tomography (CT) imaging as a method for assessing tumor-induced angiogenesis. MATERIALS AND METHODS: Functional CT imaging was mathematically analyzed for 14 renal cell carcinomas by means of two-compartment modeling using a computer-discretization approach. The model incorporated diffusible kinetics of contrast medium including leakage from the capillary to the extravascular compartment and back-flux to the capillary compartment. The correlations between functional CT parameters [relative blood volume (rbv), permeability 1 (Pm1), and permeability 2 (Pm2)] and histopathological markers of angiogenesis [microvessel density (MVD) and vascular endothelial growth factor (VEGF)] were statistically analyzed. RESULTS: The modeling was successfully performed, showing similarity between the mathematically simulated curve and the measured time-density curve. There were significant linear correlations between MVD grade and Pm1 (r = 0.841, P = 0.001) and between VEGF grade and Pm2 (r = 0.804, P = 0.005) by Pearson's correlation coefficient. CONCLUSION: This method may be a useful tool for the assessment of tumor-induced angiogenesis. PMID- 18509722 TI - Relation between cancer cellularity and apparent diffusion coefficient values using diffusion-weighted magnetic resonance imaging in breast cancer. AB - PURPOSE: The purpose of this study was to examine the relation between cancer cellularity and the apparent diffusion coefficient (ADC) value using diffusion weighted magnetic resonance imaging in breast cancer. MATERIALS AND METHODS: The subjects were 27 women who had undergone operation for breast cancer. There were 27 breast cancer lesions, 24 of which were invasive ductal carcinoma (IDC) and 3 of which were noninvasive ductal carcinoma (NIDC). RESULTS: The mean ADC values of IDC, NIDC, and normal breasts were 1.07 +/- 0.19 .10(-3), 1.42 +/- 0.17 .10( 3), and 1.96 +/- 0.21 .10(-3) mm(2)/s, respectively. The mean ADC values of IDC and NIDC were significantly different from that of normal breasts (P < 0.001 each). The mean ADC values were also significantly different between IDC and NIDC (P < 0.001). There was no correlation between the ADC value and cancer cellularity. CONCLUSION: The mean ADC values for breast cancer were significantly different from that of normal breasts. The mean ADC value for breast cancer did not significantly correlate with cancer cellularity but did correlate with histological types. PMID- 18509723 TI - Hyperacute stroke patients and catheter thrombolysis therapy: correlation between computed tomography perfusion maps and final infarction. AB - PURPOSE: We investigated the correlation between abnormal perfusion areas by computed tomography perfusion (CTP) study of hyperacute stroke patients and the final infarction areas after intraarterial catheter thrombolysis. MATERIALS AND METHODS: CTP study using the box-modulation transfer function (box-MTF) method based on the deconvolution analysis method was performed in 22 hyperacute stroke patients. Ischemic lesions were immediately treated with catheter thrombolysis after CTP study. Among them, nine patients with middle cerebral artery (MCA) occlusion were investigated regarding correlations of the size of the prolonged mean transit time (MTT) area, the decreased cerebral blood volume (CBV) area, and the final infarction area. RESULTS: Using the box-MTF method, the prolonged MTT area was almost identical to the final infarction area in the case of catheter thrombolysis failure. The decreased CBV areas resulted in infarction or hemorrhage, irrespective of the outcome of recanalization after catheter thrombolysis. CONCLUSION: The prolonged MTT areas, detected by the box-MTF method of CTP in hyperacute stroke patients, included the area of true prolonged MTT and the tracer delay. The prolonged MTT area was almost identical to the final infarction area when recanalization failed. We believe that a tracer delay area also indicates infarction in cases of thrombolysis failure. PMID- 18509724 TI - Validation of the use of calibration factors between the iodine concentration and the computed tomography number measured outside the objects for estimation of iodine concentration inside the objects: phantom experiment. AB - PURPOSE: The aim of this study was to validate the use of a calibration factor measured outside the object for estimating the iodine concentration inside the object to improve the accuracy of the quantitative contrast-enhanced computed tomography (CT). MATERIALS AND METHODS: Several known concentrations (0, 6, 9, and 12 mg I/ml) of iodine contrast material (CM) samples were placed inside and outside cylindrical acrylic phantoms of two sizes and were imaged under various combinations of the tube voltages and currents (kV/mAs-80/200, 100/200, 120/200, 140/200) to obtain K factors. The K factors were compared between the phantoms and among the tube voltages. Each CM concentration was estimated from the CT number using the K factor measured outside the phantom. RESULTS: The K factors varied between the phantoms or among the tube voltages (P < 0.05). Although there were statistically significant variations in K factors among the different regions in a phantom, the mean variation coefficient was 3%-4%. The mean error of the estimated concentration was -5.5%. CONCLUSION: The CM concentration should be accurately estimated at the region within a patient's body using the K factor measured at the surface of the body regardless of body size and tube voltage. PMID- 18509725 TI - Low-grade fibromyxoid sarcoma of the small bowel mesentery: computed tomography and magnetic resonance imaging findings. AB - Low-grade fibromyxoid sarcoma (LGFMS) is a rare tumor that commonly arises in the lower extremities but rarely in the mesentery. We report computed tomography (CT) and magnetic resonance imaging (MRI) findings of LGFMS of the small bowel mesentery. On CT, the mass was composed of two components. One component, on its right side, appeared to have isointense attenuation relative to muscle, whereas the other component, on its left side, appeared to have low attenuation. On MRI the mass on the right side showed hypointensity similar to muscle on both T1-and T2-weighted images as well as mostly slight enhancement on contrast-enhanced T1 weighted images. On the other hand, the mass on the left side showed relative hypointensity on T1-weighted images and hyperintensity on T2-weighted images as well as intense enhancement on contrast-enhanced T1-weighted images, suggesting that the tumor contained myxoid tissue. The myxoid area of LGFMS may have a tendency to reveal intense enhancement on contrast-enhanced images. PMID- 18509726 TI - Pharyngitis of infectious mononucleosis: computed tomography findings. AB - Two women presented with sore throat and fever. Their symptoms were not alleviated by antibiotics. Cervical computed tomography (CT) with contrast enhancement demonstrated enlargement of predominant posterior cervical lymph nodes and streaky heterogeneous tonsils with interspersed low attenuation. They were diagnosed as having infectious mononucleosis by their laboratory data. Thus, when radiologists encounter these CT findings of pharyngitis that is not alleviated by antibiotic therapy, infectious mononucleosis should be considered in the differential diagnosis. PMID- 18509727 TI - Occupational social class, risk factors and cardiovascular disease incidence in men and women: a prospective study in the European Prospective Investigation of Cancer and Nutrition in Norfolk (EPIC-Norfolk) cohort. AB - OBJECTIVES: To investigate the association between occupational social class and cardiovascular disease (CVD) incidence, and the extent to which classical and lifestyle risk factors explain such relationships, and if any differences persist after 65 years of age. DESIGN, SETTING AND PARTICIPANTS: Prospective population study of 22,478 men and women aged 39-79 years living in the general community in Norfolk, United Kingdom, recruited using general practice age-sex registers in 1993-1997 and followed up for total mortality to 2006. MAIN RESULTS: In both men and women an inverse relationship was observed between social class and CVD incidence, with a relative risk of social class V compared to I of 1.90 in men (95% CI 1.47 to 2.47, P < 0.001) and 1.90 in women (95% CI 1.45 to 2.49, P < 0.001). Adjusting for classical and lifestyle risk factors (age, smoking, BMI, systolic blood pressure, total blood cholesterol, history of diabetes, physical activity, weekly alcohol intake and plasma vitamin C levels) had little effect in men; the relative risk of social class V compared to I of 1.70 (95% CI 1.31 to 2.22, P < 0.001), while there was some attenuation seen in women, relative risk of social class V compared to I of 1.56 (95% CI 1.18 to 2.05, P = 0.011). The association persisted in men and women aged > or =65 years. CONCLUSIONS: Some but not all of the socioeconomic differential in CVD incidence can be explained by potentially modifiable classical and lifestyle risk factors. Low social class remains a risk factor for CVD after age 65 years. Further understanding of the mechanisms underlying the association is needed if we are to reduce inequalities in health. PMID- 18509728 TI - The association of the metabolic syndrome with QTc interval in NHANES III. AB - PURPOSE: To examine the relationship of metabolic syndrome with corrected QT interval duration. METHODS: In a cross-sectional analysis of NHANES III, a representative sample of the adult US population, 3,495 individuals aged > or =40 years were categorized as having metabolic syndrome and not having metabolic syndrome as defined by ATP III. QT interval was measured from the standard 12 lead electrocardiogram. RESULTS: A weighted multi-linear regression analysis adjusted for age, gender, serum calcium, and potassium showed that metabolic syndrome was significantly associated with a corrected QT interval in milliseconds. The adjusted Beta coefficient and its corresponding standard error were (4.48 milliseconds, 0.95 milliseconds, P < 0.01). CONCLUSION: These data indicate that metabolic syndrome is independently associated with a corrected QT interval duration. This study calls for careful ECG monitoring among persons with metabolic syndrome for early detection of a long corrected QT interval in order to prevent severe and often fatal arrhythmias. PMID- 18509729 TI - Health-economic burden of obesity in Europe. AB - Although overweight and obesity have long been recognised as major risk factors for a number of chronic diseases, lifestyle developments have led to substantial increases in bodyweight worldwide. In addition to their negative effects on health and quality of life, obesity and associated comorbidities may have a considerable impact on healthcare expenditures. The aim of this systematic review was to summarise cost estimates and compare costs attributable to obesity across different European countries. A structured search in MEDLINE, EMBASE, and all EBM Reviews was conducted to identify relevant literature. Two researchers independently assessed publications according to pre-defined inclusion criteria and with regard to study methodology. Costs attributable to obesity were extracted from the included studies and calculated relative to country-specific gross domestic income. Out of 797 publications that met our search criteria, 13 studies investigating 10 Western European countries were determined to be relevant and included in our review. Obesity-related healthcare burdens of up to 10.4 billion euros were found. Reported relative economic burdens ranged from 0.09% to 0.61% of each country's gross domestic product (GDP). Obesity appears to be responsible for a substantial economic burden in many European countries, and the costs identified in the available studies presumably reflect conservative estimates. There remains a great need for prospective and standardised studies to provide more accurate estimates of costs for all European countries. PMID- 18509730 TI - Birth spacing and maternal risk of invasive epithelial ovarian cancer in a Swedish nationwide cohort. AB - OBJECTIVE: Pregnancies reduce the risk of ovarian cancer, and among multiparous women, levels of circulating progesterone might be higher during pregnancies with wider birth spacing. We hypothesized that childbirth with wider birth spacing might reduce maternal risk of invasive epithelial ovarian cancer more than births with narrower spacing. METHODS: We conducted a case-control study nested in a nationwide cohort of Swedish women from 1961 to 2001. We selected five individually age-matched controls for each case of invasive epithelial ovarian cancer, and analysis for the effect of birth spacing was performed for 5,341 cases and 29,047 controls. We applied unconditional logistic regression analyses adjusting for age, ages at childbirth, educational level, area of residence, and gender of offspring. RESULTS: Relative risk of invasive epithelial ovarian cancer associated with each one-year increase in average birth spacing is 1.00 (95% CI = 0.98-1.01) among all women and 0.99 (0.98-1.01) among those born before 1935 and less likely to have used oral contraceptives. Further analyses on the biparous and triparous women did not find a consistent association between birth spacing and the risk of ovarian cancer. CONCLUSIONS: Birth spacing is unlikely to be a major determinant underlying the protective effects of childbirth on ovarian cancer risk. PMID- 18509731 TI - Infertility, treatment of infertility, and the risk of breast cancer among women with BRCA1 and BRCA2 mutations: a case-control study. AB - BACKGROUND: Women with a breast cancer susceptibility gene 1 (BRCA1) or breast cancer susceptibility gene 2 (BRCA2) mutation are at increased risk for developing breast and ovarian cancer. Various reproductive and hormonal factors have been shown to modify the risk of breast cancer. These studies suggest that estrogen exposure and deprivation are important in the etiology of hereditary cancer. Many patients are interested in the possibility of an adverse effect of fertility treatment on breast cancer risk. It is important to evaluate whether or not infertility per se or exposure to fertility medications increase the risk of breast cancer in genetically predisposed women. METHODS: We conducted a matched case-control study of 1,380 pairs of women with a BRCA1 or BRCA2 mutation to determine if a history of infertility, the use of fertility medications, or undergoing in vitro fertilization (IVF) were associated with and increased the risk of breast cancer. RESULTS: Sixteen percent of the study subjects reported having experienced a fertility problem and 4% had used a fertility medication. Women who had used a fertility medication were not at significantly increased risk of breast cancer (odds ratio [OR] = 1.21; 95% confidence interval [CI] = 0.81-1.82) compared to non-users. Furthermore, there was no risk associated with a history of use of a fertility medication when the subjects were stratified by parity: (OR = 1.29; 95% CI = 0.83-2.01 for nulliparous women and OR = 0.81; 95% CI = 0.30-2.22 for parous women). CONCLUSIONS: The results of this study suggest that the use of fertility medications does not adversely affect the risk of breast cancer among BRCA mutation carriers. Given the small sizes of the exposed subgroups, these findings should be interpreted with caution and confirmatory studies are required. PMID- 18509732 TI - Obstetric history and birth characteristics and Wilms tumor: a report from the Children's Oncology Group. AB - Previous epidemiologic studies have suggested that various pregnancy and birth characteristics may be associated with Wilms tumor, a childhood kidney tumor. We evaluated obstetric events and birth characteristics in relation to Wilms tumor using data from a large North American case-control study. Mothers of 521 children with Wilms tumor and 517 controls, frequency matched on child's age and geographic region, provided information about their labor and delivery history and their children's birth characteristics through a detailed computer-assisted telephone interviews. Most obstetric factors were not associated with Wilms tumor, but modest associations were observed for labor induction (OR: 1.4, 95% Confidence Interval (CI): 1.1, 1.8), prenatal vaginal infection (OR: 1.8, 95% CI: 1.2, 2.8), and upper respiratory infection (OR: 1.5, 95% CI: 1.0, 2.4). Low (<2500 g) and high (>4500 g) birth weight and preterm delivery (<37 weeks completed gestation) were associated with an elevated risk of Wilms tumor, as was neonatal respiratory problems. The association for high birth weight was present only among children with perilobar nephrogenic rests (OR: 2.1, 95% CI: 1.2, 3.9), possibly distinguishing a specific association among a biologically distinct subgroup of Wilms tumor cases. The results of this large study did not support many of the earlier findings of smaller studies. However, additional investigations of the effects of certain obstetric and birth characteristics among more refined tumor subgroups may further our understanding of these factors in relation to Wilms tumor. PMID- 18509733 TI - Solid phase extractive preconcentration of silver from aqueous samples. AB - N,N-dibutyl-N1-benzoylthiourea (DBBT) impregnated onto a polymeric matrix, Amberlite XAD-16 was prepared. The separation and enrichment of Ag(I) from solution was investigated. Effective extraction conditions were optimized in column methods prior to determination by atomic absorption spectrometry. The optimum pH range for quantitative adsorption is 2-5. Quantitative recovery of Ag was achieved by stripping with 1 mol L(-1) thiourea in 1 mol L(-1) HCl. The sorption capacity of resin is 0.115 mmol Ag+ g(-1) resin. The relative standard deviation and detection limit was 3.1% for 1 microg Ag+ mL(-1) solution and 0.11 microg L(-1), respectively. The method was used for the determination of silver in geological water samples. PMID- 18509734 TI - Characterization and prediction of meandering channel migration in the GIS environment: a case study of the Sabine River in the USA. AB - This study focused on the prediction of a 22 km meandering channel migration of the Sabine River between the states of Texas and Louisiana. The meander characteristics of 12 bends, identified from seven orthophotos taken between 1974 and 2004, were acquired in a GIS environment. Based on that earlier years' data acquisition, channel prediction was performed for the two years 1996 and 2004 using least squares estimation and linear extrapolations, yielding a satisfactory agreement with the observations (the median predicted and observed migration rates were 3.1 and 3.6 [m/year], respectively). The best-predicted migration rate was found to be associated with the longest orthophoto-recorded interval. The study confirmed that channel migration is strongly correlated with bend curvature and that the maximum migration rate of the bend corresponded to a radius of curvature [bend radius (R(C))/channel width (W(C))] of 2.5. In tight bends of a smaller radius of curvature than 1.6, secondary flow scouring near the bend apex increases bend curvature. The stability index of the dimensionless bend radius was determined to be 2.45. Overall, this study proves the effectiveness of least squares estimation with historical orthophotography for characterization of meandering channel migration. PMID- 18509735 TI - Fluoride in groundwater, Varaha River Basin, Visakhapatnam District, Andhra Pradesh, India. AB - Excess intake of fluoride through drinking water causes fluorosis on human beings in many States of the country (India), including Andhra Pradesh. Groundwater quality in the Varaha River Basin located in the Visakhapatnam District of Andhra Pradesh has been studied, with reference to fluoride content, for its possible sources for implementing appropriate management measures, according to the controlling mechanism of fluoride concentration in the groundwater. The area occupied by the river basin is underlain by the Precambrian Eastern Ghats, over which the Recent sediments occur. Results of the chemical data of the groundwater suggest that the considerable number of groundwater samples show fluoride content greater than that of the safe limit prescribed for drinking purpose. Statistical analysis shows that the fluoride has a good positive relation, with pH and bicarbonate. This indicates an alkaline environment, as a dominant controlling mechanism for leaching of fluoride from the source material. Other supplementary factors responsible for the occurrence of fluoride in the groundwater are evapotranspiration, long contact time of water with the aquifer material, and agricultural fertilizers. A lack of correlation between fluoride and chloride, and a high positive correlation between fluoride and bicarbonate indicate recharge of the aquifer by the river water. However, the higher concentration of fluoride observed in the groundwater in some locations indicates insufficient dilution by the river water. That means the natural dilution did not perform more effectively. Hence, the study emphasizes the need for surface water management structures, with people's participation, for getting more effective results. PMID- 18509736 TI - Fungal flora in indoor and outdoor air of different residential houses in Tekirdag City (Turkey): seasonal distribution and relationship with climatic factors. AB - This study was investigated the density and monthly distribution of indoor and outdoor microfungi in six different residential houses in Tekirdag City through the exposure of Petri dishes containing Rose-Bengal Streptomycin Agar media. Samples were collected in 1-month intervals over a period of 12 months between March, 2001, and February, 2002. We used 432 Petri dishes and counted a total of 4,205 microfungi colonies, 1,790 from indoor air and 2,415 from outdoor air. As a result, 42 species belonging to 12 genera were identified. The most frequent fungal genera were Penicillium (28.61%), Cladosporium (16.08%) and Alternaria (15.98%). While Penicillium (40.61%) and Cladosporium (15.92%) were the dominant genera of indoor air, Alternaria (20.62%) and Penicillium (19.71%) were isolated most frequently from outdoor air (Table 3). Alternaria citri (10.15%) and Penicillium brevicompactum (10.15%) were found to be the most frequent among the 42 identified species. While P. brevicompactum (19.55%) and Aspergillus niger (6.37%) were the most frequent indoor species, A. citri (13.37%) and Cladosporium cladosporioides (8.20%) were the most frequent outdoor species. Linear Regression Analysis was applied to determine whether or not there was a relationship between the number of colonies of isolated fungal genera and meteorological factors during the research period. Correlations between the presence of Aspergillus and temperature, relative humidity, duration of sunny periods and agents of air pollution such as SO(2) and PM were statistically significant. No significant correlations, however, were found between other fungal genera and environmental variables. PMID- 18509737 TI - Monitoring the condition of natural resources in US national parks. AB - The National Park Service has developed a long-term ecological monitoring program for 32 ecoregional networks containing more than 270 parks with significant natural resources. The monitoring program assists park managers in developing a broad-based understanding of the status and trends of park resources as a basis for making decisions and working with other agencies and the public for the long term protection of park ecosystems. We found that the basic steps involved in planning and designing a long-term ecological monitoring program were the same for a range of ecological systems including coral reefs, deserts, arctic tundra, prairie grasslands, caves, and tropical rainforests. These steps involve (1) clearly defining goals and objectives, (2) compiling and summarizing existing information, (3) developing conceptual models, (4) prioritizing and selecting indicators, (5) developing an overall sampling design, (6) developing monitoring protocols, and (7) establishing data management, analysis, and reporting procedures. The broad-based, scientifically sound information obtained through this systems-based monitoring program will have multiple applications for management decision-making, research, education, and promoting public understanding of park resources. When combined with an effective education program, monitoring results can contribute not only to park issues, but also to larger quality-of-life issues that affect surrounding communities and can contribute significantly to the environmental health of the nation. PMID- 18509738 TI - Typology, classification and management issues of Greek lakes: implication of the Water Framework Directive (2000/60/EC). AB - The European Water Framework Directive 2000/60/EC, requires the determination of ecological status in European waterbodies, based primarily on biological indicators, and that minimum good ecological quality is obtained by 2015. We used background morphometric, hydrologic, physico-chemical and biological data, in order to assess the typology of the main Greek lakes. Correlation analysis and multivariate analysis (Principal Component Analysis) were applied for the data statistical elaboration. Lakes were classified according to the characteristics suggested in the Directive and specified in the Annex II. Statistical analysis revealed significant correlations between the lake variables as well as the grouping of the Greek lakes according to their abiotic characteristics. Lake water uses, in most cases competed each other, were identified and management practices were also recorded. We suggest that the future conservation efforts should focus on a new holistic management philosophy taking into consideration the catchment's management. PMID- 18509739 TI - Effect of biotin supplementation on hoof health and ceramide composition in dairy cattle. AB - The effect of biotin supplementation on various foot lesions and hoof ceramide composition of toe (wall) and sole portions of hooves was studied in crossbred dairy cattle. Biotin supplementation was done for five months in 14 cattle at a farm and the other 14 animals kept as control. A significant decline was observed in heel erosions and sole avulsions along with total disappearance of white line fissures and double soles in the biotin supplemented cattle resulting in decrease in the overall disease score. Thin layer chromatographs of the hoof lipids revealed 11 types of ceramides in sole lipids and 6 types of ceramides in toe (wall) lipids. The ceramides were typed and identified according to their Rf values. A qualitative increase in the density of thin layer chromatographs of sole lipids was observed in biotin supplemented cattle whereas a non-significant difference in density of thin layer chromatographs of toe lipids was observed after supplementation of biotin. PMID- 18509740 TI - Phylogenetic background of attaching and effacing Escherichia coli isolates from animals. AB - Detection and distribution of eae gene in forty-four attaching and effacing Escherichia coli (AEEC) strains of animal origin were investigated. Association of distinct intimin alleles with phylogenetic background were assessed among strains in comparison with different serogroups. Phylogenetic analysis showed that 31 EHEC/eae+ STEC strains belong to groups A, B1 and E, 13 EPEC strains segregated in B1 and B2. Moreover, group A possessed the eae gamma2/theta type, group B1 the eae beta1, eae kappa, eae zeta, and eae epsilon types, group B2 the eae alpha1, eae alpha2 and eae iota types, while the group E possessed the eae gamma1 type. The presence of numerous eae-types show that EPEC and EHEC/eae+ STEC tested have a high genetic homology within each phylogenetic group. PMID- 18509741 TI - Repeat prescribing in primary care: a prescription study. AB - OBJECTIVE: There is limited knowledge on repeat prescribing of different drug groups in cases where the physician does not see the patient. This study aimed to investigate the frequency of repeat prescribing of different drug groups both with and without consultation. METHOD: The study was a nationwide prescription database study performed in Finnish primary care in 2001. Each physician in a sample of 400 physicians was asked to report all patient contacts over a specified period of five consecutive working days once over a period of six months, and another sample of 400 physicians did the same for the next six months. Copies of physicians' prescriptions were collected and data on the drugs and type of prescriptions were recorded. Repeat prescriptions issued with and without a consultation were included. RESULTS: About 19% of all prescriptions were issued without direct contact between the physician and the patient. In the different drug groups, the range was from 2% of antibiotics to 57% of antipsychotics. Psychotropics including hypnotics and tranquillizers, cardiovascular drugs and drugs for elderly patients were most often issued without consultation. CONCLUSION: Repeat prescribing without consultation is common especially for elderly patients and users of psychotropics and cardiovascular drugs, and this may contribute to suboptimal therapy. PMID- 18509742 TI - Twenty-four-month alpha-galactosidase A replacement therapy in Fabry disease has only minimal effects on symptoms and cardiovascular parameters. AB - Fabry disease is an X-linked lysosomal storage disease caused by deficiency of alpha-galactosidase A enzyme activity. Decreased enzyme activity leads to accumulation of glycosphingolipids in different tissues including endothelial cells and smooth-muscle cells and cardiomyocytes, and cardiovascular complications are common in the disease. Since 2001, specific enzyme replacement therapy (ERT) with alpha-galactosidase A has been available. It has been reported to improve clinical symptoms and quality of life. However, limited and controversial data on its efficacy to cardiac involvement have been published. Nine patients (5 male) with Fabry disease were included in an open-label prospective follow-up study of 24-month ERT. Comprehensive cardiovascular evaluation was performed by MRI, stress echocardiography and quality of life assessment. Plasma globotriaosylceramide decreased from 6.2 to 1.4 microg/ml during ERT (p<0.05). The only other measured parameters that changed significantly were resting heart rate that decreased from 79 to 67 bpm (p<0.01) and end-systolic volume that decreased by 12.4 ml (p<0.05). The other parameters consisting of quality of life, self-estimated cardiovascular condition, diastolic function, exercise capacity, ECG parameters, ejection fraction and ventricular mass did not change. ERT has only minimal effect on symptoms and cardiovascular morphology and function in Fabry disease. Therefore, effective conventional medical therapy is still of major importance in Fabry disease. Larger ERT studies are warranted, especially in women, to solve current open questions, such as the age at which ERT should be started, optimal dosage and intervals between infusions. Furthermore, longer follow-up studies are needed to assess the effects of ERT on prognosis. PMID- 18509743 TI - Clinical characterization of cardiovascular abnormalities associated with feline mucopolysaccharidosis I and VI. AB - OBJECTIVE: The purpose of this study was to define the cardiovascular abnormalities present in young and adult cats affected with the lysosomal storage diseases mucopolysaccharidosis (MPS) I and MPS VI. METHOD: Eighteen cats affected with MPS I and 10 cats affected with MPS VI were evaluated by physical examination, electrocardiography and echocardiography. Electrocardiography (ECG) was performed on all MPS I and 9 of the MPS VI cats. Twelve unaffected cats underwent complete examinations for comparison purposes. RESULTS: No cardiovascular abnormalities were noted on physical examination. Measured ECG intervals were normal in affected cats; however, sinus arrhythmia was noted more frequently than in the unaffected cats. Significant echocardiographic abnormalities included aortic valve thickening, regurgitation and aortic root dilation. Significant mitral valve thickening was also noted. The severity of changes increased in older affected cats. CONCLUSION: As affected animals increased in age, more cardiac abnormalities were found with increasing severity. Significant lesions included the mitral and aortic valves and ascending aorta, but myocardial changes were not recognized. MPS I and MPS VI cats have similar cardiovascular findings to those seen in children and constitute important models for testing new MPS therapies. PMID- 18509744 TI - Renal tubular function in children with tyrosinaemia type I treated with nitisinone. AB - BACKGROUND: Tyrosinaemia type I (TTI) is an inherited deficiency in the enzyme fumarylacetoacetate hydrolase and is frequently complicated by renal tubular dysfunction which may persist in some patients after hepatic transplantation. Nitisinone has revolutionized the management of TTI but its effect on renal tubular dysfunction has not been described in a large cohort of patients. AIMS: To document the incidence and progression of renal tubular dysfunction in children with TTI treated with nitisinone at a single centre. SUBJECTS: Twenty one patients with TTI from a single centre were treated with nitisinone for at least 12 months. Median age at first treatment was 17 weeks (range 1 week to 27 months). Nine patients (43%) presented in acute liver failure, seven (33%) had a chronic presentation and five (24%) were detected pre-clinically. METHODS: A retrospective case analysis of plasma phosphate, urinary protein/creatinine ratio and tubular reabsorption of phosphate was performed for all patients as markers of tubular function. Renal ultrasounds were examined for evidence of nephrocalcinosis and where available, skeletal radiographs for rickets. RESULTS: All patients had biochemical evidence of renal tubular dysfunction at presentation. After nitisinone and dietary treatment were started, all three markers normalized within one year. Four children had clinical rickets at presentation (which improved), of whom one had nephrocalcinosis, which did not reverse on nitisinone. No child redeveloped tubular dysfunction after commencing nitisinone. All patients with TTI had evidence of tubular dysfunction at presentation and in all cases this resolved with nitisinone and dietary control. CONCLUSION: The tubulopathy associated with TTI is reversible. PMID- 18509746 TI - Identification of a mutation in exon 27 of the RB1 gene associated with incomplete penetrance retinoblastoma. AB - Retinoblastoma (Rb) is initiated by germline mutations in the RB1 gene. Up to date, no mutation was identified in exons 26 and 27. We have identified a 2 bp frameshift insertion in exon 27 of the RB1 gene (RBg.177008_177009dup) in a boy with unilateral Rb and his healthy father that has occurred de novo on the allele transmitted by the father's father. RT-PCR showed that the mutant +2 bp transcript is present in RNA from peripheral leukocytes after short-term culture. The level of the mutant transcript was low compared to the normal transcript indicating abnormal expression of the variant allele. The mutant transcript was further reduced after puromycin treatment suggesting that NMD is not involved. Although oncogenic mutations in the terminal exons of the RB1 gene are rare molecular testing is important as those terminal mutations can be associated with incomplete penetrance and cause high recurrence risk in family members. PMID- 18509747 TI - Bone marrow transplantation results in donor-derived hepatocytes in an animal model of inherited cholestatic liver disease. AB - Cell transplantation is a potential therapy for acquired or inherited liver diseases. Donor-derived hepatocytes (DDH) have been found in humans and mice after bone marrow transplantation (BMT) but with highly variable frequencies in different disease models. To test the effect of liver repopulation after BMT in inherited cholestatic liver diseases, spgp (sister of P-glycoprotein, or bile salt export pump, abcb11) knockout mice, a model for human progressive intrahepatic cholestasis type 2 with defects in excreting bile salts across the hepatocyte canalicular membrane, were transplanted with bone marrow cells from enhanced green fluorescent protein (EGFP) transgenic donor mice after lethal irradiation. One to 6 months later, scattered EGFP-positive DDHs with positive spgp staining were observed in the liver. These hepatocytes had been incorporated into hepatic plates and stained positively with hepatocyte-specific marker albumin. RT-PCR for the spgp gene revealed positive expression in the liver of sgsp knockout mice that had received the transplant. Bile acid analysis of bile samples showed that these mice also had higher levels of total biliary bile acid and taurocholic acid concentration than knockout mice without transplantation, indicating that BMT partially improved biliary bile acid secretion. Our results indicate that bone marrow cells could serve as a potential source for restoration of hepatic functions in chronic metabolic liver disease. PMID- 18509745 TI - Management of non-neuronopathic Gaucher disease with special reference to pregnancy, splenectomy, bisphosphonate therapy, use of biomarkers and bone disease monitoring. AB - Enzyme replacement was introduced as treatment for non-neuronopathic Gaucher disease more than 15 years ago. To ensure the best use of this costly ultra orphan agent, a systematic disease management approach has been proposed by an international panel; this includes the development, by consensus, of achievable treatment goals. Here we critically review these goals and monitoring guidelines and incorporate emerging experience of the disease in the therapeutic era, as well as contemporary clinical research. This review makes recommendations related specifically to the management of pregnancy; the appropriate use of splenectomy and bisphosphonate treatment; the relevance of biochemical markers to disease monitoring; and the use of semi-quantitative methods for assessing bone marrow infiltration. In addition, we identify key areas for development, including the requirement for a validated index of disease severity; the need to correlate widely used biomarkers with long-term disease outcomes, and the desirability of establishing agreed standards for monitoring of bone disease particularly in infants and children with Gaucher disease. PMID- 18509748 TI - Effect of small interference RNA targeting HIF-1alpha mediated by rAAV combined L: -ascorbate on pancreatic tumors in athymic mice. AB - To study the effect of recombinant adeno-associated virus (rAAV) vector bearing small inference RNA (siRNA) targeting hypoxia inducible factor 1alpha (HIF 1alpha) combined L: -ascorbate on pancreatic tumors in athymic mice primarily. A cassette encoding siRNA targeting HIF-1alpha mediated by rAAV was constructed, giving rAAV-siHIF. In vitro, rAAV-hrGFP, rAAV-siHIF and L: -ascorbate which were used alone or in combination were delivered to exponentially growing MiaPaCa2 cells. Then, we examined the expression of HIF-1alpha mRNA and protein, the secretion of VEGF in MiaPaCa2 cells under hypoxic condition with Real-time PCR, Western Blot, ELISA, respectively. In vivo, MiaPaCa2 cells were inoculated subcutaneously on the back of nude mice. Nude mice with xenograft tumor were randomly divided into equal groups and were injected with rAAV-hrGFP or rAAV siHIF or were fed with L: -ascorbate. Then, we measured the size of tumor every 3 days and drew a tumor growth curve. After 30 days, all mice were sacrificed and the tumors were dissected. At last, we examined the expression of HIF-1alpha, VEGF and CD34 by immunohistochemistry and counted micro-vessel density (MVD). In vitro, we found that rAAV-siHIF could inhibit the expression of HIF-1alpha mRNA and protein in MiaPaCa2 human pancreatic cancer cells but L: -ascorbate could only restrain the expression of HIF-1alpha protein. Moreover, rAAV-siHIF and L: ascorbate could all inhibit the secretion of vascular VEGF. In vivo, we found that rAAV-siHIF could inhibit the growth of nude mice xenograft tumor and the expression of HIF-1alpha and VEGF and MVD while L: -ascorbate can only inhibit the growth of xenograft tumor in the early and middle stage. These results suggest that rAAV-siHIF and L: -ascorbate can inhibit the growth of nude mice xenograft tumor and HIF-1alpha could be a target of pancreatic cancer genetic and pharmacological therapy. PMID- 18509749 TI - Do differences in accuracy of vital records and hospital discharge data between physician and nurse-midwife attended births matter? PMID- 18509750 TI - Reasons for ineffective contraceptive use antedating adolescent pregnancies: part 2: a proxy for childbearing intentions. AB - PURPOSE: Compare the relationship between childbearing intentions, maternal behaviors, and pregnancy outcomes in a group of early/middle adolescents versus a group of late adolescents (specifically high school seniors, high school graduates, and GED certificate recipients). METHODS: The reasons given by a racially/ethnically diverse group of 1,568 pregnant 13-18 year olds for not using contraception were used to classify their pregnancies as intended or unintended. Proportion comparison tests and stepwise logistic regression analyses were used to study the relationship between childbearing intentions, maternal behaviors, and pregnancy outcomes. RESULTS: Regardless of age, adolescents who intended to become pregnant conceived in an objectively more hospitable and supportive childbearing milieu than those who conceived unintentionally. This is evidenced by their greater likelihood of having goals compatible with adolescent childbearing, cohabitation with the father of the child, and living in a non chaotic environment. However, pregnancy planning was not associated with improved compliance with preventive health care recommendations during gestation nor with infant outcomes. As such, the consequences among adolescents with intended pregnancies were negative, as evidenced by a higher rate of smoking, STDs late in gestation, school dropout, and repeat conception. CONCLUSIONS: Like adults, adolescents with intended pregnancies conceived in an objectively more supportive environment than their counterparts with unintended pregnancies. However, this advantage did not translate into better support, healthier maternal behavior during gestation, or improved pregnancy outcomes. PMID- 18509752 TI - Qualities of symbolic play among children with autism: a social-developmental perspective. AB - We hypothesized that the qualities of play shown by children with autism reflect their impoverished experience of identifying with other people's attitudes and moving among person-anchored perspectives. On this basis, we predicted their play should manifest a relative lack of the social-developmental hallmarks that typify creative symbolic functioning. We videotaped the spontaneous and modelled symbolic play of matched groups of children with and without autism. The two groups were similar in the mechanics of play, for example in making one thing stand for another and using materials flexibly. By contrast, and as predicted, children with autism were rated as showing less playful pretend involving self conscious awareness of pretending, investment in the symbolic meanings given to play materials, creativity, and fun. PMID- 18509753 TI - A comparative analysis of sexual risk characteristics of Black men who have sex with men or with men and women. AB - Many behavioral studies of Black men fail to differentiate between those who have sex exclusively with men (MSM) and those who have sex with both men and women (MSMW). Respondent-driven sampling was used to recruit a total of 1,154 Black MSM and MSMW in New York City and Philadelphia. In descriptive analyses, MSMW and MSM were compared on several demographic, health, and behavioral risk correlates using chi-square tests. Differences in prevalence of unprotected anal intercourse (UAI) between these two groups were examined in two multivariate logistic regressions. Age, country of birth, self-identified sexual orientation, experience of being forced to have sex, self-reported HIV status, exchange sex for money/food/ drug, and drug use in the past 3 months were significantly associated with either insertive or receptive UAI in the past 3 months. The strongest correlate of either insertive or receptive UAI among both groups of men was engaging in exchange sex. Differences between MSMW and MSM were found in the areas of forced sexual experiences, disclosure of same sex behavior, and history of being arrested or incarcerated. Findings from our study highlight the need for specific HIV prevention interventions targeting Black MSMW as distinguished from Black MSM. PMID- 18509754 TI - The involvement of the Toll-like receptor family in ovulation. AB - INTRODUCTION: Ovulation is similar to an inflammatory response and is associated with increased production of prostaglandins as well as local growth regulatory factors. However, the expression and function of innate immune cell-related genes in non-immune cells within the ovary has been reported recently and provides a novel and important regulatory system during ovulation. DISCUSSION: Several members of the Toll-like receptor (TLR) surveillance system are expressed in granulosa cells and cumulus cells. These receptors can be activated by pathogens as well as endogenous ligands leading to the induction and release of potent cytokines and chemokines from cumulus cells. CONCLUSION: These inflammatory factors exert potent effects on cumulus cell-oocyte expansion, ovulation, transport and fertilization indicating that ovulation is a more complex immune inflammatory process than previously recognized. PMID- 18509755 TI - Depressed adolescents and comorbid psychiatric disorders: are there differences in the presentation of depression? AB - Patterns and correlates of comorbidity, as well as differences in manifest depressive profiles were investigated in a sample of depressed adolescents. A sub sample of the youth were characterized as belonging to either a Pure depression group, an Internalizing group (depression and co-occurring internalizing disorders), or an Externalizing group (depression and co-occurring externalizing disorders). Item response theory (IRT) and differential item functioning (DIF) were used to assess whether the depressed adolescents from the different comorbidity groups presented with different depressive symptoms. Results indicated that the comorbidity groups were meaningfully distinct in terms of psychosocial correlates as well as showed differences in depressive symptom profiles as informed by DIF analyses. In particular, the comorbidity groups differed in terms of presentation of psychomotor changes and cognitive impairments. Implications for assessment are discussed. PMID- 18509756 TI - Personality and electrodermal response lability: an interpretation. AB - Electrodermal response (EDR) lability is a psychophysiological trait reflecting stable individual differences in electrodermal activation as indexed by frequency measures of phasic EDR activity. There is no consistent evidence that EDR lability reflects dispositional or clinical anxiety. However, EDR lability appears to be related to individual differences in the overt expression of emotional and antagonistic impulses. Greater EDR lability is associated with a relatively undemonstrative and agreeable disposition, whereas greater EDR stability is associated with a relatively expressive and antagonistic disposition. The inverse relationship between EDR lability and the expression of emotional and antagonistic impulses suggests that EDR lability may reflect individual differences in the effortful control of such expression. This hypothesis is consistent with cognitive effort interpretations of phasic EDR activity, with evidence of the sensitivity of phasic EDR activity to capacity demanding tasks, and with evidence of reduced spare capacity among EDR labile individuals under cognitive challenge. Individual differences in effortful self control may explain the association of greater EDR lability with essential hypertension and greater EDR stability with forms of antisocial behavior. PMID- 18509757 TI - Circulating steroid hormone concentrations in postmenopausal women in relation to body size and composition. AB - Steroid hormones are associated with the risk of postmenopausal breast cancer and evidence suggests that increased concentrations of oestrogens from peripheral aromatisation in adipose tissue partly explains the association between body mass index (BMI) and risk of postmenopausal breast cancer. This study examined the associations between circulating concentrations of steroid hormones and anthropometric measurements in a sample of naturally postmenopausal women from the Melbourne Collaborative Cohort Study, not using hormone replacement therapy. We measured plasma concentration of total oestradiol, oestrone sulphate, dehydroepiandrosterone sulphate, androstenedione, testosterone and sex hormone binding globulin (SHBG) and calculated concentration of free oestradiol. Body measurements included height, weight, BMI, waist circumference, fat mass and fat free mass, the last two estimated by bioelectrical impedance analysis. BMI was positively associated with both oestrogens and androgens and negatively with SHBG. Fat mass was the principal measure responsible for the association observed between body size and total oestradiol. The associations between oestrone sulphate and androgens and body size were mainly with waist circumference. The associations between oestrogens and body size were close to null for the first 6 years since menopause and became positive thereafter. Our results are compatible with the hypothesis that after the menopause excess fat mass increases oestrogen concentrations through the peripheral aromatisation of androgens in adipose tissue. This effect requires around 6 years to be detectable by way of circulating steroid hormone levels. PMID- 18509759 TI - Importance of travel in domestically acquired typhoid fever infections: opportunities for prevention and early detection. AB - Approximately 25% of Salmonella typhi infections in the US occur among nontravelers. Two S. typhi infections in a major US metropolitan city acquired domestically in 2005 were epidemiologically linked to a S. typhi-infected Haitian traveler through their congregation meetings. This investigation highlighted the importance of integrating multiple methods of obtaining epidemiologic information, including laboratory evidence and multiple individual and group interviews. Physicians should consider typhoid fever in their differential diagnosis in communities with close ties to endemic areas. Education of communities whose residents travel regularly to typhoid-endemic areas can reduce infection and transmission risk. PMID- 18509758 TI - A comprehensive meta-analysis of Triple P-Positive Parenting Program using hierarchical linear modeling: effectiveness and moderating variables. AB - A meta-analysis encompassing all studies evaluating the impact of the Triple P Positive Parenting Program on parent and child outcome measures was conducted in an effort to identify variables that moderate the program's effectiveness. Hierarchical linear models (HLM) with three levels of data were employed to analyze effect sizes. The results (N=55 studies) indicate that Triple P causes positive changes in parenting skills, child problem behavior and parental well being in the small to moderate range, varying as a function of the intensity of the intervention. The most salient findings of variables moderating the interventions' impact were larger effects found on parent report as compared to observational measures and more improvement associated with more intensive formats and initially more distressed families. Sample characteristics (e.g., child's age, being a boy) and methodological features (e.g., study quality) exhibited different degrees of predictive power. The analysis clearly identified several strengths of the Triple P system, most importantly its ability to effect meaningful improvement in parents and children. Some limitations pertain to the small evidence-base of certain formats of Triple P and the lack of follow-up data beyond 3 years after the intervention. Many of the present findings may be relevant to other evidence-based parenting programs. PMID- 18509760 TI - Hidden homicide increases in the USA, 1999-2005. AB - Prior to 1999, dramatic fluctuations in homicide rates were driven by changes in the rates of firearm homicide among men aged 15-24. Since 2000, the overall homicide rate has appeared stable, masking any changes in population subgroups. We analyzed recent trends in homicide rates by weapon, age, race, gender, state, and urbanization to determine whether the risk of victimization increased substantially during 1999-2005 for demographic subgroups. The analysis of WISQARS data and Wonder data from Centers for Disease Control and Prevention revealed no trend in the homicide rate nationally between 1999 and 2005; this obscured large increases in firearm homicide rates among black men aged 25-44 and among white men aged 25-34. Between 1999 and 2005, for ages 25-44 combined, the increase for black men was 31% compared with 12% for white men. Significant increases among men aged 25-44 occurred in Alabama, California, Michigan, Minnesota, Nebraska, Nevada, New Jersey, Ohio, Pennsylvania, Texas, and Washington. The firearm homicide rate increased the most in large central metropolitan areas (+32%) and large fringe metropolitan areas (+30%) for men aged 25-44. We conclude that the recent, unrecognized increases in firearm homicide among men aged 25-44, especially black men, in large metropolitan areas merit the attention of policymakers. PMID- 18509761 TI - Identification of intracellular signaling cascades mediating the PACAP-induced increase in guinea pig cardiac neuron excitability. AB - The pituitary adenylate cyclase-activating polypeptide (PACAP) increases excitability of guinea pig cardiac neurons, an effect mediated through activation of PAC1 receptors. The signaling cascades that couple activation of the PAC1 receptor to alterations in membrane ionic conductances responsible for the PACAP effect are unknown. Intracellular recordings were made from neurons in kinase inhibitor-treated cardiac ganglia preparations to determine which of the intracellular cascades activated by PAC1 receptor stimulation mediate the PACAP effect. In control cells, long depolarizing-current steps elicited one to three action potentials. In contrast, during the application of 10 nM PACAP, depolarizing-current pulses elicited multiple action potential firing (greater than or equal to five action potentials) in 79% of the neurons. Pretreatment with an adenylyl cyclase inhibitor, SQ 22536 (100 microM), suppressed the PACAP induced increase in excitability, whereas the presence of U-73122 (10 microM), a potent phospholipase C (PLC) inhibitor, had no effect. Thus, the activation of adenylyl cyclase, but not PLC, was a critical step mediating the PACAP effect. Pretreatment with H-89 (1 microM), a protein kinase A inhibitor, and PD 98059 (50 microM), a MEK kinase inhibitor, also significantly blunted the PACAP-induced increase in excitability. Furthermore, treatment with forskolin (5 microM), an activator of adenylyl cyclase, or exposure to the cell-permeable cyclic adenosine monophosphate (cAMP) analogue, 8-bromo-cAMP (1 mM), partially recapitulated the effect of PACAP on excitability. We conclude that the activation of signaling cascades downstream of cAMP mediate the PACAP-induced increase in cardiac neuron excitability. PMID- 18509762 TI - Maxadilan, the PAC1 receptor, and leishmaniasis. AB - Maxadilan is a vasodilator peptide isolated from sand fly salivary glands. The vasodilator effects of maxadilan are mediated by the PAC1 receptor, although maxadilan and PACAP do not share sequence homology. Sand flies are the vector of the parasitic disease leishmaniasis. The peptide aids the sand fly in obtaining a blood meal while enhancing the infectivity of leishmania parasites transmitted by this arthropod vector. Aspects of maxadilan, PAC1, and leishmaniasis are discussed. PMID- 18509763 TI - Cerebrovascular manifestations of pheochromocytoma and the implications of a missed diagnosis. AB - BACKGROUND AND PURPOSE: Cerebrovascular complications are a rare manifestation of pheochromocytoma. We report a case of pheochromocytoma presenting initially with reversible posterior leukoencephalopathy and subsequently a cerebral infarction. A patient with a prior history of episodic hypertension and reversible posterior leukoencephalopathy presented with headache, palpitations, and weakness of the right leg. A magnetic resonance angiogram revealed narrowing of the basilar and right middle cerebral artery. She was diagnosed as having presumed vasculitis and underwent a conventional cerebral angiogram during which she developed a hypertensive crisis and worsening neurological deficit. Further investigations revealed the presence of a pheochromocytoma, which was subsequently resected. CONCLUSIONS: It is important to be cognizant of the possibility of pheochromocytoma in patients presenting with hypertension and cerebrovascular manifestations, as commonly used investigations (e.g. catheter angiography) and treatment modalities (e.g. beta-blockers to treat hypertension, corticosteroids to treat suspected vasculitis, etc.) can lead to life threatening complications. PMID- 18509764 TI - The chemical pathway to primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a female predominant chronic disease of autoimmune pathogenesis and unknown etiology, although data suggest that genetic predisposition and environmental factors concur to its onset. Among nongenetic factors, several lines of evidence spanning from geoepidemiology to experimental findings support the role of xenobiotics, i.e., chemicals that are capable to induce molecular mimicry through cross reactivity. Indeed, specific xenobiotics are hypothesized to substitute lipoic acid residues on PBC-specific autoepitopes thus triggering autoimmunity. This is supported by data obtained with patient sera reactivities as well as animal models. The scenario is further complicated by the possibility that xenobiotic-metabolizing bacteria might also play a role. We will review the available evidence in this intriguing and rapidly growing field of research and critically discuss its potential implications. PMID- 18509765 TI - P. aeruginosa Biofilms in CF Infection. AB - Pseudomonas aeruginosa is an opportunistic pathogen of immunocompromised hosts. In cystic fibrosis (CF), P. aeruginosa causes acute and chronic lung infections that result in significant morbidity and mortality. P. aeruginosa possesses several traits that contribute to its ability to colonize and persist in acute and chronic infections. These include high resistance to antimicrobials, ability to form biofilms, plethora of virulence products, and metabolic versatility. In P. aeruginosa, a cell-to-cell communication process termed quorum sensing (QS) regulates many of these factors that contribute to its pathogenesis. Recent evidence suggests that the CF lung environment presents a specialized niche for P. aeruginosa. The relationship of P. aeruginosa QS, biofilm formation, and the CF lung environment is discussed. PMID- 18509766 TI - Unbalanced jumping translocation involving 3q in myeloproliferative disease. AB - An 87-year-old woman was diagnosed with unclassified myeloproliferative disease having an acquired jumping translocation with the long arm of chromosome 3 translocating to the short arm telomeric region of chromosome 8 (major clone) and the long arm telomeric region of chromosome 10 (minor clone). Each abnormal clone was also associated with an extra copy of chromosome 8. Although there was no evidence of transformation to an acute leukemia, the patient deteriorated until her demise 7 months after disease presentation. There have been fewer than 70 cases of acquired jumping translocations reported in the literature. To our knowledge, this is the first acquired jumping translocation case to be reported in a patient with myeloproliferative disease. PMID- 18509767 TI - [Levofloxacin in the treatment of nosocomial infection in critically ill patients]. AB - INTRODUCTION: Levofloxacin (LVX) is one of the most frequently used antibiotics in critical patients admitted to Spanish Intensive Care Units (ICU). Their use in community-acquired infections has been widely documented, while it is less frequent and known in nosocomial infections (NI). OBJECTIVE: To describe the indications and utilization patterns of LVX in the treatment of NI in patients admitted to Spanish ICU. MATERIAL AND METHODS: Open-label, retrospective, observational and multicenter study. All patients admitted to ICU and who were being treated for NI with LVX in the years 2004-2005 were included. A case report form (CRF) was drawn up and included demographic, infection, treatment, infectious process and patient development variables. NI-dependent LVX usage was described. A logistical regression analysis was carried out in order to identify the variables associated with a satisfactory response. Results are expressed by means of the odds ratio and a 95% confidence interval. RESULTS: A total of 949 patients who were given LVX for the treatment of 1,103 NI were recruited in 87 ICU: 460 (41.7%) with non-mechanical ventilation associated pneumonia, 256 (23.2 %) mechanical-ventilation associated pneumonia, 107 (9.7 %) with primary or vascular catheter-related bacteremia, 47 (4.3 %) with urethral catheter-related urinary infections, 42 (3.8%) with organspace or deep surgical infections and 191 (17.3%) who had other types of infection. An APACHE II upon admission of 19.6 (SD: 8) and severe sepsis or septic shock systemic response in 50.4% of all cases. On 776 (82.7%) occasions treatment was initiated on an empirical basis and in 589 (62.1%) cases the dose of choice was of 0.5 g/ 12 h, with a mean duration of 9 days. In 738 (77.8 %) patients, LVX was used in association with other antibiotics. The clinical response by treatment end was rated as satisfactory in 67.4 % of all NI. Factors related to a non-satisfactory response were as follows: APACHE II (OR: 1.05; 95% CI: 1.028-1.078); septic shock (OR: 2.62; 95 % CI: 1.623 4.219); the requirement for changes in treatment due to poor clinical progress (OR: 66.67; 95% CI: 15.384-250), the presence of non-covered microorganisms (OR: 6.58; 95% CI: 3.663-11.765), the appearance of new resistant pathogens (OR: 6.94; 95 % CI: 2.445- 19.608) or the diagnosis of a new infection (OR: 3.68; 95% CI: 1.504-8.929); solid neoplasm (OR: 1.98; 95% CI: 1.156-3.899); chronic liver disease (OR: 3.11; 95 % CI: 1.429-8.475) and the absence of etiology confirmation (OR: 2.39; 95 % CI: 1.624-3.510). One or more adverse events which were possibly or probably related to the use of LVX were detected in 104 (11.0%) patients. Total intra-ICU mortality amounted to 26.1%, while the accumulated in-hospital mortality was 33.8%. CONCLUSIONS: LVX is a common therapeutic option in the treatment of nosocomial infections in critical patients. It is predominantly used in an empirical manner, at a dose of 0.5 g every 12 hours and in combination with other antibiotics. PMID- 18509768 TI - [Methicillin-resistant Staphylococcus aureus bacteremia. Predictor factors for an isolate with a vancomycin minimal inhibitory concentration > or =2 mg/l]. AB - A greater rate of treatment failures with vancomycin in methicillin-resistant Staphylococcus aureus (MRSA) bacteremia has been reported recently when the minimum inhibitory concentration (MIC) is > or =2 mg/l. This study has aimed to evaluate if there are clinical and/or epidemiological factors that predict isolation of a MRSA strain with MIC of vancomycin of > or =2 mg/L in the bacteremia episodes collected during a 15 year period (January 1991 to December 2005) in a tertiary urban hospital. During the study period, a total of 478 episodes of MRSA bacteremia were studied prospectively. The following clinical variables were recorded for each one: age, gender, comorbidity, previous administration of vancomycin or another antibiotic, prognosis of baseline diseases, bacteremia focus, shock, empiric antibiotic received and mortality. The MIC of vancomycin of 419 strains (88%) was determined with the E-test. In 216 (52%) of the isolations the MIC of vancomycin was 1.50 mg/L, in 110 (26%) of the cases it was < or =1 mg/l and in 93 (22%) 2 mg/l. Uni-and multivariate analyses were made, comparing the clinical variables of the patients infected by strains with MIC of vancomycin > or =2 mg/l regarding the MIC strains < or =1 mg/l. In the last 3 years of the study (2003-2005) the proportion of the strains with MIC of vancomycin > or =2 mg/l was significantly greater than those isolated with MIC < or = 1 mg/L (44 % vs 3 %; p<0.001). In the multivariate analysis, the only clinical characteristic associated independently to the isolation of a strain with MIC > or =2 mg/l was the nosocomial-acquired infection OR (95 % CI): 1.94 (1.04-3.63); p=0.04. Although the isolation of a MRSA strain with MIC of vancomycin > or =2 mg/l is more frequent in the nosocomial-acquired bacteremia episodes, in the clinical practice, it is not a useful predictive parameter because the frequency of isolation of these strains in the community is also high. PMID- 18509769 TI - [Anidulafungin]. AB - Anidulafungin is a new echinocandin antifungal agent recently approved in Spain by the Spanish Drug Agency. As other echinocandins, it inhibits a selective target, 1,3- beta-D-glucan synthesis, a major structural component of the fungal cell wall which is not present in mammalian cells, this avoiding toxicity problems. It has fungicidal activity against many Candida spp., including fluconazole-resistant, and fungistatic activity against other yeast and moulds such as Aspergillus spp. Clinical trials have shown non-inferiority of anidulafungin to fluconazole for invasive, including candidemia, and non-invasive Candida infections. It is well-tolerated, and no drug-related serious adverse events have been reported. Anidulafungin, which has a very long half life, is slowly degraded by human peptidases and proteases and has a low drug-drug interaction profile based on its lack of interaction with the cytochrome P450 system. Thus, dosing adjustments of anidulafungin based on age, gender, body weight, disease status, concomitant therapy or renal or hepatic insufficiency is not necessary. As it does not interact with amphotericin B and voriconazole, cyclosporine, tacrolimus and other drugs, it can be used in combination with other antifungal agents and co-administered with immunosuppressant drugs. It is generally well-tolerated in clinical trials. Its most frequent adverse events are nausea, vomiting, moderate diarrhea, transient elevation of hepatic enzymes and headache. Some of the patients have mild, passing reactions such as facial blushing, nausea and dyspnea related with rapid intravenous perfusion. Its antifungal activity, clinical efficacy, safety profile, and pharmacokinetic characteristics make it a suitable alternative antifungal compound for therapy of mucosal candidiasis, candidemia and invasive candidiasis, above all in patients with some degree of renal and hepatic insufficiency. PMID- 18509770 TI - [Clinical relevance of bacterial resistance: a historical approach (1982-2007)]. AB - Bacterial resistance is currently one of the most important problems of infectious pathology. The relation between in vitro and in vivo bacterial resistance is not always well defined because therapeutic failure is also related to other factors (pharmacokinetics and pharmacodynamics). In addition, there are disagreements between the in vitro and in vivo activity of several antimicrobials (especially ciprofloxacin) due to their low bactericidal activity. In infections due to ciprofloxacin susceptible S. pyogenes, S. aureus, S. epidermidis, E. faecalis, E. coli producing extended spectrum beta-lactamase (ESBL), K. pneumoniae and E. cloacae their clinical use is not associated to cure because of the development of resistances that are induced during the antibiotic treatment. Ceftazidime in infections due to susceptible strains of K. pneumoniae and E. cloacae and ceftriaxone in infections due to methicillin susceptible S. aureus also do not have a good correlation between in vitro and in vivo results due to their low bactericidal activity and to the development of resistances during treatment. The main clinical impact of resistant bacteria is related to the failure of empirical treatments, which is associated to a higher mortality, especially in severe infections with methicillin-resistant S. aureus, Enterobacteriae ESBL and multiresistant P. aeruginosa and A. baumannii. One of the main risk factors for the development of bacterial resistances is the increase of the consumption of several antibiotics. The development of protocols agreed upon by consensus may decrease the impact of bacterial resistances. The knowledge of the previous use of antibiotics is an especially relevant issue to suspect that an infection might be due to resistant bacteria. Resistant pathogens are a severe problem in the clinical setting and the question is of such a complexity that it requires a multidisciplinary effort that involves the different professionals of the Internal Medicine-Infectious Diseases, Microbiology, Pharmacology, Pharmacy and Preventive Medicine Departments and hospital directors and that results in unified and protocolized actions regarding the clinical and therapeutical approach for the management of severely infected patients. PMID- 18509771 TI - Isolation of the first metallo-beta-lactamase producing Klebsiella pneumoniae in Lebanon. AB - INTRODUCTION: A 58 year-old man was admitted to the Saint Joseph Hospital-Raymond and Aida Najjar polyclinic in Beirut on July 17, 2007 to undergo surgery for a moderately differentiated colonic adenocarcinoma (T3N0). Following several discharges and re-admissions, an extended spectrum beta-lactamase (ESBL) producing Escherichia coli susceptible to imipenem was isolated. The patient was put on imipenem and metronidazole. Three weeks later, imipenem (IMP) resistant Klebsiella pneumoniae was isolated. METHODS AND RESULTS: The antimicrobial susceptibility profile of the imipenem-resistant Klebsiella pneumoniae strain and related minimum inhibitory concentrations of antibiotics were determined. Hydrolysis of IMP was evaluated and production of metallo-beta-lactamase (MBL) was detected by a double disk-synergy test, ethylene diamine tetraacetic acid (EDTA) inhibited the imipenemase activity, whereas clavulanate and tazobactam did not, this suggesting the production of a metallo-beta-lactamase. Isoelectric focusing analysis was performed and indicated the presence of a cefotaximase (blaCTX-M-15). Polymerase chain reaction (PCR) was used and detected the presence of blaIMP-1 and blaCTX-M genes. CONCLUSIONS: During the last decade, many hospital outbreaks caused by ESBL-producing Enterobacteriaceae spp. have been reported in Lebanon. To our knowledge, this is the first report of a clinical isolate of K. pneumoniae producing an MBL in Lebanon. PMID- 18509772 TI - [Recommendations of antifungal treatment in patients with low grade immunosuppression]. AB - Because of the relevance that the systemic mycoses has acquired in non-highly immunocompromised patients, the treatment difficulties they have due to the increase of the non-albicans Candida species and the need to have a better and more rational use of the new antifungal agents (voriconazole, posaconazole, caspofungin, anidulafungin and micafungin), an experts' panel on infectious diseases in representation of the Spanish Society of Chemotherapy, Spanish Society of Internal Medicine, and Spanish Society of Pneumology and Thoracic Surgery has met in order to make a few recommendations based on the scientific evidence in an effort to improve their efficiency. PMID- 18509773 TI - American Chemical Society--235th National Meeting. Part 1. PMID- 18509774 TI - American Chemical Society--235th National Meeting. Part 2. PMID- 18509775 TI - American Chemical Society--235th National Meeting. Part 3. PMID- 18509776 TI - Experimental Biology 2008. Today's research: tomorrow's health. PMID- 18509777 TI - Antiviral research--21st international conference. PMID- 18509778 TI - Clinical Microbiology and Infectious Diseases (ECCMID)--18th European Congress. New antimicrobial agents: selected poster presentations. PMID- 18509779 TI - Clinical Microbiology and Infectious Diseases (ECCMID)--18th European Congress. Drug resistance among Gram-negative and Gram-positive bacteria. PMID- 18509780 TI - American Association for Cancer Research--99th Annual Meeting. Coverage of AKT and MET pathways and antibody-based therapeutics. PMID- 18509781 TI - American Association for Cancer Research--99th Annual Meeting. Clinical and preclinical data on promising therapeutics. PMID- 18509782 TI - Society for biomolecular sciences--14th Annual Conference and Exhibition. Enhancing research productivity: quality tools, leads and candidates. PMID- 18509783 TI - High-content assays in oncology drug discovery: opportunities and challenges. AB - High-content screening (HCS), a process that combines fluorescence microscopic imaging and automated image analysis, has had a significant impact on drug discovery since its introduction in the mid 1990s. The application of HCS within pharmaceutical drug discovery has become widespread, notably within oncology drug discovery. The trends, challenges and considerations for HCS that affect the successful and pragmatic implementation of this process in drug discovery will be outlined. PMID- 18509784 TI - Gender issues and the pharmacotherapy of substance abuse. AB - In the last 25 years, the different disciplines and professions that are involved in managing the issue of substance abuse have recognized that gender-linked factors influence patterns of substance abuse and response to treatment. In 2007, the US National Institute on Drug Abuse (NIDA) concluded that studies of substance abuse should routinely address gender issues. In the field of drug development, gender issues historically have not been an issue of high priority. By outlining the types of evidence that underpin the view of the NIDA and providing evidence of the widespread existence of gender-linked effects from pharmacotherapies (both those marketed and under development), this feature review proposes that drug developers should embrace the opinion of the NIDA regarding gender and substance abuse. To avoid false conclusions, the issue of gender should become a higher priority during the collection and analysis of data on pharmacotherapies. Some gender differences will have more clinical significance than others, but any difference related to gender has the potential to complicate clinical trials and other studies. PMID- 18509785 TI - Eligen insulin--a system for the oral delivery of insulin for diabetes. AB - The oral administration of insulin is difficult to achieve because the large peptide hormone is poorly absorbed and is subjected to enzymatic and acidic degradation in the stomach. Emisphere Technologies Inc is developing formulations of insulin co-administered with a drug delivery agent. With the proprietary Eligen technology employed, the carrier agent appears to form a conformational complex with insulin that can protect against degradation and facilitate the absorption of the hormone through the intestinal wall. In animal studies and phase I clinical trials, dosing with Eligen insulin led to a rapid elevation of plasma insulin and subsequent decrease in plasma glucose levels; the onset of activity was more rapid and insulin concentrations were higher with Eligen insulin than with injected insulin. Formulations of Eligen insulin have been well tolerated in all clinical trials performed to date. In a phase II clinical trial in patients with type 2 diabetes, Eligen insulin in combination with metformin failed to achieve significant superior glycemic control over treatment with metformin alone. Eligen oral insulin formulations may have potential as prandial insulin therapies in patients with either type 1 or type 2 diabetes; however, no clinical trials for such treatments appeared to be ongoing at the time of publication. PMID- 18509786 TI - Pharmacological closure of patent ductus arteriosus: effects on pulse pressure and on endothelin-1 and vasopressin excretion. AB - Widened pulse pressure is a classic sign of significant left-to-right shunting patent ductus arteriosus (PDA), but little evidence supports this statement in the early life of premature infants with respiratory distress syndrome (RDS) needing nonsteroidal anti-inflammatory drugs (NSAIDs), the pharmacological treatment for PDA. Pulse pressure and urinary endothelin-1 (ET-1) and arginine vasopressin (AVP) vasoactive factors involved in the transitional circulation were measured before and after the NSAIDs treatment of 46 RDS premature infants receiving either ibuprofen (n = 22) or indomethacin (n = 24), with 28 responders and 18 nonresponders to the first NSAIDs course. We found that following pharmacological PDA closure, systolic and diastolic blood pressure significantly increased, maintaining a stable pulse pressure. However, when pharmacological closure failed, the trend (nonsignificant) was for a more consistent increase in systolic than in diastolic blood pressure, which determined a statistically significant widening pulse pressure. In addition, urinary ET-1 excretion rates decreased significantly after PDA closure, whereas persistent more aggressive pharmacological therapy failed. Urinary AVP excretion rates decreased insignificantly after therapy, uninfluenced by the efficacy of the drugs. We concluded that widened pulse pressure is a clinical sign of failed PDA pharmacological closure in RDS premature infants. ET-1 levels remain elevated when NSAIDs fail to interrupt left-to-right PDA shunting that complicates recovery from RDS. PMID- 18509787 TI - Pregnancy outcomes during an era of aggressive management for intrahepatic cholestasis of pregnancy. AB - Our objective was to examine whether delivery at 37 weeks of gestation alters adverse pregnancy outcomes in Latina patients with intrahepatic cholestasis of pregnancy (ICP). We conducted a retrospective chart review of Latina patients who delivered at our institution coded with ICP between 2000 and 2007. During this time period it was our practice to offer delivery to patients with ICP at 37 weeks of gestation. Subjects were classified into three groups according to total bile acid (TBA) concentration: < 20 micromol/L (mild ICP), > or = 20 micromol/L and < 40 micromol/L (moderate ICP), and > or = 40 micromol/L (severe ICP). Meconium passage was observed in no births in patients with mild IC, but was found in 18% of deliveries with moderate/severe ICP. The risk of meconium passage increased linearly, with a 19.7% increased risk for each 10 mumol/L increase in TBA concentration ( P = 0.001). There was no association with higher TBA concentration and other adverse outcomes. There was no difference in adverse outcomes between moderate and severe ICP. We concluded that in our Latina population with ICP, an association existed between meconium passage and moderate/severe ICP. Delivering at 37 weeks was associated with a low risk of adverse outcomes due to ICP among all patients, including those with higher TBA concentrations. PMID- 18509788 TI - Protease inhibitor therapy and fetal growth potential in HIV-positive women. AB - Our objective was to test if protease inhibitors (PIs) increase the incidence of fetal growth restriction (FGR). Human immunodeficiency (HIV)-seropositive women were studied. At birth the neonatal weight percentile was assigned by predicted growth potential (GP), accounting for race, parity, weight, height, gestational age, birthweight, and gender (Gardosi, 1992). FGR was defined as GP < 10% percentile. Maternal age, CD4 count, viral load, weight gain, prenatal care, tobacco, alcohol, substance abuse, and PI use were related to FGR using chi square and multiple regression analysis. Ninety-three of 191 women received PI. In these, FGR occurred in 27 (29%) compared with 15 (15.3%) in the non-PI group ( P = 0.02). Maternal CD4 count ( P < 0.0001) was the primary determinant, and smoking ( P = 0.037) was an independent cofactor for FGR (Nagelkerke r2 = 0.24). Twenty-six of 82 (31.7%) smokers had FGR, versus 16 of 109 (14.7%) of nonsmokers (odds ratio, 2.69; 95% confidence interval, 1.33 to 5.46; P = 0.005). After exclusion of the CD4 count, PI became a cofactor for FGR ( P = 0.021 and Nagelkerke r2 = 0.104). We concluded that maternal HIV status and smoking determine the risk for FGR. Although PIs increase the risk for FGR, this effect appears to depend on maternal disease severity. PMID- 18509789 TI - Neonatal arterial thrombosis at birth: case report and literature review. AB - Neonatal thromboembolism is rare and in most cases iatrogenic from indwelling central catheters, peripheral arterial lines, and umbilical lines. It often requires urgent intervention to restore perfusion and to avoid morbidity and even mortality. Very few case reports of neonatal arterial thrombosis at birth are described in the literature. We present the case of a full-term infant noted to have marked left arm swelling at birth. Doppler ultrasound of the arm demonstrated a large thrombus in the left subclavian, axillary, and brachial arteries. He underwent left arm fasciotomy and anticoagulant therapy with good recovery. We present this case with a brief review of the literature. PMID- 18509790 TI - Impingement and entrapment syndromes. PMID- 18509791 TI - External impingement of the shoulder. AB - The relationship between external shoulder impingement and rotator cuff disease has been the subject of much research, but the theories of cause and effect remain controversial. Patients with symptoms of external impingement are referred for imaging to identify bony abnormalities of the coracoacromial arch and associated bursal and rotator cuff disease. Attempts have been made to identify objective imaging criteria that confirm the diagnosis of impingement, but at present external impingement remains primarily a clinical diagnosis. Therapeutic management varies from rehabilitation with physiotherapy to surgical procedures aimed at decompressing the subacromial space and repairing rotator cuff tears. This article reviews the relevant anatomy, biomechanics, and theories of external impingement, the role of imaging in the diagnosis of external impingement and rotator cuff disease, and implications upon management. PMID- 18509792 TI - Internal impingement syndromes of the shoulder. AB - The internal impingement syndromes are a group of conditions that result from the impingement of the soft tissues of the rotator cuff and joint capsule on the glenoid or between the glenoid and the humerus. They should not be confused with conditions where impingement of cuff and bursa occur on the structures of the coracoacromial arch, so-called external impingement. Some controversy surrounds the true etiology of the soft tissue injuries seen in internal impingement syndromes and whether they are truly the result of impingement. Internal impingement seems to be a normal physiological occurrence with the shoulder in certain positions. Imaging findings in these conditions include undersurface partial-thickness cuff tears, superior labral pathology, and bone changes. PMID- 18509793 TI - Femoroacetabular impingement: presentation, diagnosis, and management. AB - Femoroacetabular impingement (FAI) is an often overlooked cause of hip pain in patients of all ages. The clinical signs and symptoms, multimodality imaging findings, treatment options, intraoperative findings, and expected outcomes for patients with FAI are illustrated and discussed. PMID- 18509794 TI - Anterior ankle impingement syndromes. AB - Ankle impingement syndromes are painful conditions that may complicate ankle trauma and are characterized by chronic, progressive pain, swelling, and limitation of movement. These disorders are subclassified according to anatomical location about the tibiotalar joint. This article reviews the various forms of anterior ankle impingement, detailing the unique clinical features, anatomical considerations, pathoetiology, and imaging findings for each. PMID- 18509795 TI - Posterior impingement syndromes of the ankle. AB - Acute, or repetitive, compression of the posterior structures of the ankle may lead to posterior ankle impingement (PAI) syndrome, posteromedial ankle impingement (PoMI) syndrome, or Haglund's syndrome. The etiology of each of these conditions is quite different. Variations in posterior ankle osseous and soft tissue anatomy contribute to the etiology of PAI and Haglund's syndromes. The presence of an os trigonum or Stieda process is classically associated with PAI syndrome, whereas a prominent posterosuperior tubercle of the os calcis or Haglund's deformity is the osseous predisposing factor in Haglund's syndrome. PoMI has no defined predisposing anatomical variants but typically follows an inversion-supination injury of the ankle joint. This article discusses the biomechanics, clinical features, imaging, and management of each of these conditions. Magnetic resonance imaging (MRI) provides the optimal tool in posterior ankle assessment, and this review focuses on the MRI findings of each of the conditions just listed. PMID- 18509796 TI - Imaging of entrapment and compressive neuropathies. AB - Although the subject of entrapment and compressive neuropathies is huge, with dedicated textbooks on the subject, this article attempts to provide an up-to date overview of the role of imaging in the diagnosis of nerve entrapment and compression syndromes. Entrapment and compressive neuropathies are a group of distinct syndromes secondary to physical constriction or irritation affecting peripheral nerves at specific anatomical sites in the body. Most nerve entrapment and compressive syndromes derive from an injury to the neurovascular components in a narrow anatomical passage. Because of their etiological diversity, which includes pressure, angulation, stretch, and friction, the pathophysiology of individual nerve entrapment syndromes differs widely. Neuropathy can result in considerable morbidity. Although the mainstay of achieving diagnosis is with clinical acumen and electrophysiological investigations, the increasing use of modern high-resolution imaging studies is of particular value in confirming physical findings and enabling determination of the extent of injury. Knowledge and familiarity of pertinent anatomy and appropriate choice of imaging modality is important for the radiologist to allow accurate interpretation of site and etiology of nerve entrapment and compression as well as ascertaining possible alternative diagnoses. PMID- 18509798 TI - [Introduction of a critical incident reporting system in a surgical university clinic. What can be achieved in a short term?]. AB - BACKGROUND AND OBJECTIVE: "Critical incident reporting systems" (CIRS) are voluntary systems which, within the framework of risk management, provide pointers to the type and origin of critical incidents (including "errors"). The interest in introducing a CIRS is great, but whether it can fulfill its promises remains to be clarified. The aim of this study was to answer the question of whether an CIRS in a structured form would be acceptable to staff and whether it would be suitable for introducing targeted measures for achieving improvements. METHODS: The introduction of a CIRS in the Department of Visceral, Thoracic and Vascular Surgery proceeded according to the recommendations of the Action Alliance for Patients' Safety (Aktionsbundnis fur Patientensicherheit e.V.). RESULTS: During a period of 13 months we received a total of 96 reports, 29.3% from various levels of carers/nurses, while 35.4% were from doctors. 40.6% of the incidents had been observed by the reporting person, in 38.5% of cases the reporting person had been involved in the incident and in 12.5% this person had helped in dealing with the incident. 38.5% of the reported critical incidents occurred between 06.00 and 12.00, while 34.4% occurred between 12:00 and 18:00 o/c. In 32.3% of cases the estimated duration of dealing with the incident was under four hours and between four and eight hours in 26%. During the first year of this study 12 actions were started and continued in consequence of an analysis of the reported critical incidents. CONCLUSION: After one year of the study it was found that a CIRS can be reliably introduced into a surgical department in accordance with the recommendations of the Action Alliane for Patients' Safety. When introduced correctly CIRS provides valuable information, which will lead to risk reduction in surgery. PMID- 18509799 TI - [Influence of IT-supported clinical pathways on patient satisfaction at a surgical department of a university hospital]. AB - BACKGROUND: Clinical pathways (CPs) are considered to be a device of clinical process management, which describe the optimal way of a special type of patient with its diagnostic and therapeutic medical treatment. Apart from these economic aspects CPs can make a contribution to an optimization of the health quality management as well as to an improvement of medical staffs and patients satisfaction. In our hospital clinical pathways supported by information technology (IT-supported CPs) were implemented world-wide for the first time in a running Hospital Information System (SAP/i.s.h.med) and used by routine. The aim of the study was to investigate the influence of IT-supported CPs on patient satisfaction. MATERIAL AND METHODS: The patient satisfaction was examined BEFORE introduction of IT-supported CPs by standardized questionnaires in 64 Patients (45f/19m, median age: 64,9 +/- 1,24 years) and in 62 patients (38f/24m, median age: 63,3 +/- 1,49 years) AFTER introduction of IT-supported CPs by standardized questionnaires. Different CPs were selected and grouped by simple, middle and high complexity, each to benign and malignant illnesses. RESULTS: By introduction of IT-supported CPs patient satisfaction can be improved. CPs already evaluated as very good before introduction of CPs could be improved only slightly, whereas badly evaluated CPs exhibited a large optimization potential. CONCLUSION: On the one hand patient satisfaction may be improved by the introduction of CPs. On the other hand CPs- when IT-supported- do not result unavoidably in an industrial medicine. This subjective estimate of the patient contributes to a better customer-and patient-oriented quality management, whereby an appropriate optimization of the recognized deficits can be simply realized by IT-supported CPs. PMID- 18509801 TI - [Iatrogenic trauma following resuscitation measures]. AB - Resuscitation attempts require invasive iatrogenic manipulations on the patient. On the one hand, these measures are essential for successful survival of the patient, but on the other hand can damage the patient. We differentiate between frequent and rare injuries. Factors of influence are duration and intensity of the resuscitation attempts, age of the patient as well as a coagulate-inhibiting medication. PMID- 18509800 TI - [Myocardial infarction-like electrocardiogram in acute Hantavirus infection. Suspected secondary peri-myocarditis]. AB - HISTORY AND ADMISSION FINDINGS: A 19-year-old man was admitted to our hospital because of fever and exhaustion. During his hospital stay he had pain in the back and flanks. INVESTIGATIONS: The electrocardiogram (ECG) revealed ST-segment elevation in V1-V3, similar to that occurring in myocardial infarction. Renal function was decreased with proteinuria and the platelet count was decreased The symptoms and clinical findings aroused the suspicion of a Hantavirus infection, which was confirmed by antibody tests. TREATMENT AND COURSE: The patient was treated symptomatically with paracetamol. After 5 days the patient felt better and had a normal temperature. The thrombocyte count and renal function normalised after 10 days in hospital. CONCLUSION: In case of a suspected peril-/ myocarditis a Hantavirus infection should be considered as a possible cause, which should be clarified by antibody tests. PMID- 18509802 TI - [Intraductal papillary mucinous neoplasm of the pancreas]. PMID- 18509803 TI - [Emergencies in patients with implanted pacemakers or defibrillators]. PMID- 18509804 TI - [Significance of the neuraminidase inhibitor oseltamivir for the management of seasonal influenza]. PMID- 18509805 TI - [Is an elimination of infectious diseases using vaccination possible?]. PMID- 18509807 TI - The interaction between impaired acute insulin response and insulin resistance predict type 2 diabetes and impairment of fasting glucose. AB - BACKGROUND: Impaired acute insulin response (AIR) and insulin resistance (IR) are characteristics of Type 2 diabetes (T2DM). The aim was to develop risk models for T2DM and impaired fasting glucose (IFG), reflecting estimates both of AIR and IR, and of their interaction, as predictors over 20 years of follow-up. METHODS: We developed predictive models using hierarchic multiple regression analyses in a population-based cohort of 1227 men with normal fasting blood glucose at baseline (1970-73) and were reinvestigated after 10 and after 20 years. Using IVGTT variables correlated either to AIR or to IR, separate models were developed. Combined models were also estimated from which prediction scores, representing individual risk, were calculated. RESULTS: In combined models, interaction between prediction scores reflecting AIR and IR predicted T2DM and IFG. Lowest tertile of AIR and the highest tertile of IR showed a relative risk (RR) of 15.3 (95%-CI=5.58-41.84) for T2DM compared to the contrast group (high AIR and low IR). Corresponding RR for IFG was 13.23 (95%-CI=6.53-26.78). C-statistic increased from 0.76 to 0.79 (p=0.018) for T2DM and from 0.77 to 0.80 for IFG (p=0.062) taking interaction into account. Main effects of lowest tertile of AIR and highest tertile of IR versus best were: RR for T2DM, 8.80 (95%-CI=4.25-18.21) and 6.31 (95%-CI=3.26-12.21); for IFG, 9.07, (95%-CI=5.38-15.29) and 4.49 (95% CI=2.98-6-76). CONCLUSION: The interaction between low AIR and high IR revealed a high relative risk for T2DM or IFG reflecting the interplay between these factors over long time on worsening glucose tolerance and development of manifest disease. PMID- 18509808 TI - The Minimal Insomnia Symptom Scale (MISS): a brief measure of sleeping difficulties. AB - To evaluate basic psychometric properties and obtain normative values for a novel 3-item scale, the Minimal Insomnia Symptom Scale (MISS), a sleep questionnaire was sent out to a randomly selected sample of the general population, aged 20-64 years. Responses were obtained from 1075 subjects corresponding to a response rate of 78%. Results showed that MISS possessed satisfactory reliability and validity. Women scored significantly higher than men while there was no age relationship. A receiver operating characteristic curve analysis revealed that MISS was able to distinguish subjects with a clinical insomnia according to ICD 10 research criteria. The main advantage of MISS over other insomnia instruments is its brevity and ease of use. Evidence was provided for the utility of MISS in epidemiological settings. MISS also showed promise as a convenient ultra-short screening measure of insomnia in health care settings. PMID- 18509809 TI - Informal parental traffic training and children's traffic accidents. AB - The aims of the present study were (a) to assess the relationship between informal traffic training by parents and their children's involvement in traffic accidents and (b) to identify factors contributing to this relationship. The first two studies involved questionnaires on informal parental traffic education, the child's exposure to traffic and traffic-related accidents. Both studies showed that rate of accidents increased with training, particularly for outdoor training. An accident analysis indicated that most accidents involved the use of the bicycle, and that the major part of the accidents resulted in light injuries and occurred when the child was practicing the act of manoeuvring the bicycle. An interview study with 10 preschool teachers identified two quite disparate traffic education goals: emphasis on cautiousness versus emphasis on independence. The major implications of the study are that efforts in traffic training should give more emphasis to bicycle use and should be planned and carried out in cooperation with the parents. PMID- 18509810 TI - TCI : Target controlled infusion, or totally confused infusion? Call for an optimised population based pharmacokinetic model for propofol. AB - Different pharmacokinetic models for target controlled infusion (TCI) of propofol are available in the recently launched open TCI systems. There is also a compelling choice to work with either plasma- or effect-site targets. Knowledge about the clinical consequences of different alternatives is of importance. We aimed to illustrate the potential differences in the actual drug delivery/output between three present commercially available and clinically used pharmacokinetic models: the original Marsh model, which is also implemented in the Diprifusor, the "modified Marsh-" and the Schnider models. Simulations were made in the TivaTrainer program (eurosiva.com). Firstly, our standard plasma target regimen was simulated, and secondly an effect-site target of 3.5 microg/mL was chosen. Thirdly, real infusors were used for measuring the time to reach defined predicted effect-site concentrations when aiming at a plasma target of 6 microg/mL. Identical patient characteristics were used in all simulations: male, 170 cm, 70 kg, 40 years of age. Resulting predicted effect-site peak concentrations, and used bolus doses were recorded, as were the resulting plasma over-shoot, and time frames. The plasma target regimen gave predicted effect-site peaks in the different models ranging from 3.6 to 7.2 microg/mL, reached after 2(3/4) to 4 minutes. To reach the same effect-site target, the three models used bolus doses ranging from 68 to 150 mg given during 22 to 46 seconds. The predicted plasma concentration over-shoots varied from 5.0 to 13.4 microg/mL. There were obvious differences between the models in the time taken to reach defined effect-site concentrations. We observed clinically significant different results between the models. The choice of model will make a difference for the patient. To eliminate confusion - not necessarily to improve precision - we call for an optimised population based pharmacokinetic model for propofol - a consensus model! PMID- 18509811 TI - Relative proportions of serum carbamazepine and its pharmacologically active 10,11-epoxy derivative: effect of polytherapy and renal insufficiency. AB - BACKGROUND: The proposed action mechanism and pharmacological activity of carbamazepine (CBZ) and its major metabolite, carbamazepine-10,11-epoxide (CBZE), are the same. The aim of our study was the investigation of the effect of concomitant antiepileptic treatment and renal insufficiency on the relative proportions of serum CBZ and CBZE. METHODS: Serum trough steady-state CBZ and CBZE concentrations were determined by high-performance liquid chromatography (HPLC) in 140 epileptic patients treated with CBZ in monotherapy (n=100) and polytherapy with phenytoin, phenobarbital and valproate (n=40). The levels of CBZ were also determined using the Dade Behring enzyme multiplied immunoassay technique (EMIT). The glomerular filtration rate (GFR) was estimated from serum cystatin C using the Dade Behring nephelometric immunoassay. RESULTS: The CBZE/CBZ and CBZ+CBZE/CBZEMIT ratios were significantly increased in 7 cases (3 in monotherapy and 4 in polytherapy) with GFR<60 mL/min/1.73m2 in relation to the patients treated in monotherapy or polytherapy having normal or mildly decreased renal function (p<0.001). CONCLUSIONS: In patients with moderate to severe renal insufficiency the relative proportion of CBZE with respect to the parent drug is significantly increased. In these cases, the CBZ concentrations obtained using the EMIT, or other immunoassays having low CBZE cross-reactivity, may have an inadequate diagnostic efficiency. PMID- 18509812 TI - Effects of wrist splinting for Carpal Tunnel syndrome and motor nerve conduction measurements. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is one of the most common disease among the entrapment neuropathies. Wrist splinting has been conventionally used for the CTS treatment. The purposes of this study were to assess the efficacy of wrist splinting for CTS, and to evaluate the value of the motor nerve conduction measurement as a prognostic indicator for CTS. METHODS: Two hundred and fourteen hands with CTS were treated by wrist splinting, and reviewed after a mean follow up of seven months. Severity of symptoms were minimal lesions in 177 hands, intermediate lesions in 33 hands, and severe lesions in four hands. Motor nerve conduction measurement was performed in all cases before and after treatment, and distal latency (DL) and amplitude on compound muscle action potential (CMAP) from the abductor pollicis brevis (APB) muscle were analyzed. RESULTS: According to Kelly's grading of outcome, results were excellent in 41 hands, good in 110 hands, fair in 45 hands, and poor in 18 hands. Excellent or good results were obtained in 131 hands (74 percent) with minimal lesions, 20 hands (61 percent) with intermediate lesions, and in no cases with severe lesions. The ratio of excellent or good results was 79 percent in patients in whom DL of pre-treatment APB-CMAP was less than 8 milliseconds (ms), and 62 percent in patients whose DL was 8 ms or more, which showed a significant difference. In nine hands whose pre treatment APB-CMAP was unrecordable, the results were good in one hand, fair in five, and poor in three. CONCLUSIONS: Wrist splinting is most effective in cases of minimal or intermediate lesions with DL of APB-CMAP less than 8 ms. If relief of symptoms is not obtained after five months of treatment by splinting, that would be the limit of splinting. Surgical release is recommended for cases with severe lesions and with unrecordable APB-CMAP. PMID- 18509813 TI - Multiple primary malignancies involving lung: an analysis of 40 cases. AB - We aimed to assess the incidence of multiple primary malignancies in primary lung cancer patients. We retrospectively evaluated the clinical files of 1038 primary lung cancer patients diagnosed in 2004. Forty patients (3.9 %) had multiple primary malignancies. There were 34 men (85 %) and 6 women (15 %). Their mean age was 62.4 +/- 8.6 years. While 35 cases were smokers, 5 cases were nonsmokers. Tumour pathology of the lung was squamous cell carcinoma in 15 cases, adenocarcinoma in 10 cases, small cell carcinoma in 3 cases and non-small cell carcinoma in 12 cases. There were 2 primary tumours in 37 cases and 3 primary tumours in 3 cases. The first detected tumour was located in larynx in 11 cases, in genitourinary system in 9 cases, in intestine in 5 cases, in lung in 3 cases and in other organs in 12 cases. The mean interval between the first and the second tumour was 77 months with a range of 1 months to 32 years. This interval was shorter than 6 months in 4 cases. Treatment modality for the first detected tumour was surgery in 35 cases. The last primary tumour was treated with surgery in 12 cases. In conclusion, the development of multiple primary tumours is not a rare phenomenon. Patients with a malignancy should be followed for development of a second primary malignancy. The treatment of lung cancer in patients with a previous malignancy should be the same as for lung cancers presenting as the first cancer. PMID- 18509814 TI - Clinical outcome of fragment fixation for osteochondritis dissecans of the elbow. AB - BACKGROUND: The choice of surgical or non-surgical treatments for osteochondritis dissecans (OCD) of the humeral capitellum is still controversial. The purpose of this study was to assess the efficacy of fragment fixation for OCD of the humeral capitellum. METHODS: We reviewed 28 patients with OCD of the humeral capitellum after a mean follow up of 17 months. All patients were men and mean age was 14 years. Twenty-seven patients had a history of repetitive overuse of the elbow with baseball pitching, one with tennis. Mean duration of overuse of the elbow was four years. All patients had elbow pain and difficulty in throwing, with a mean duration of symptoms for 17 months. The mean arc of flexion before surgery ranged from 11 degrees to 126 degrees. Radiographs of the elbow showed a radiolucent cystic area of the humeral capitellum in one patient, a non-displaced split type fragment in 12 patients, and a slightly displaced split type fragment in 15 patients. Fragment fixation surgery was performed in all patients by lateral arthrotomy including drilling and fixation of the fragment with a double wiring technique using flexible wire or thread under direct vision. Sport activities using upper extremities were restricted for four to six months until the lesion healed in radiograph. RESULTS: Post-operatively, 25 patients had no pain and three decreased pain. Average arc of flexion was one to 132 degrees, an improvement of 16 degrees compared with the pre-operative arc. Radiographic findings showed complete healing of the lesion in 11 patients, partial healing in 12, unchanged in three, and loose body formation in two. By Tivnon's evaluation of the elbow function, results were excellent in 19 patients, good in five, fair in two, and poor in two. The ratio of complete or partial healing of the lesion was 100 percent in 16 patients in whom the thickness of the lesion was less than 9 mm on pre-operative radiograph, and 58 percent in 12 patients in whom the lesion thickness was 9 mm or more, which showed a significant difference (p<0.01). CONCLUSIONS: Fragment fixation for OCD of the humeral capitellum was effective in patients whose lesion thickness was less than 9 mm. Fixation by flexible wire or thread and revascularization by drilling for the fragment were considered to be insufficient for large lesions with a thickness of 9 mm or more. PMID- 18509815 TI - Eosinophilic granuloma arising from the pelvis in children: A report of three cases. AB - Eosinophilic granuloma (EG) is a benign tumor-like condition which is characterized by a clonal proliferation of Langerhans-type histiocytes and defined as a local form of Langerhans cell histiocytosis (LCH). The radiographic appearances of EG are quite different depending on the phase of the disease and the site of involvement. A status of EG in the bone is divided into acute and chronic phases. Radiologically acute phase of EG is difficult to differentiate from a malignant bone tumor such as Ewing's sarcoma or acute osteomyelitis. Chronic phase of EG may mimic a chronic osteomyelitis or a benign bone tumor. We report 3 children's cases of EG in the pelvis which showed quite different radiological features and clinical courses. A 6-year-old boy (Case 1) had an osteolytic lesion with slightly defined margins in the right acetabulum. A 4-year old boy (Case 2) had a radiologically similar-looking lesion in the left acetabulum. These lesions resembled radiologically chronic osteomyelitis (Brodie's abscess) or a benign bone tumor and healed spontaneously after biopsy. A 2-year-old boy (Case 3) had an osteolytic lesion with ill-defined margin in the ilium. It was difficult to differentiate from a malignant tumor such as Ewing's sarcoma, or acute osteomyelitis. The lesion became enlarged after needle biopsy. In spite of an additional curettage, the osteolytic lesion remained in the left pelvis in 1 year. Treatment for EG is controversial. Curettage of the affected site and bone grafting is usually accomplished. However, some EG heal spontaneously. It is of great importance to understand the natural course of EG and this knowledge will give us the opportunity to avoid unnecessary treatment. EG with poor osteolytic margins may progress further after biopsy. EG with well defined margins may heal spontaneously after biopsy only. PMID- 18509816 TI - Porous titanium granules for implant stability and bone regeneration - a case followed for 12 years. AB - The clinical and radiological results were excellent for each of the nine fixtures individually tested in a case followed for 12 years after implant treatment using split crest technique in combination with porous titanium granules for an augmentation of the dento-alveolar ridge in a severely resorbed maxilla. PMID- 18509817 TI - Expression of transforming growth factor-beta1 and connective tissue growth factor in the capsule in a rat immobilized knee model. AB - BACKGROUND: Contracture is a very common complication of joint immobilization in daily examination, but its cause is still unknown. A fibrotic change of the capsule is suggested to be one of the main causes of the joint contracture. The goal of this study was to analyze the expression pattern of transforming growth factor-beta1 (TGF-beta1) and connective tissue growth factor (CTGF), which are implicated in fibrosis in the capsule of a rat immobilized knee model. MATERIALS AND METHODS: We immobilized the unilateral knee joints of 66 rats in 150 degrees of flexion using a plastic plate and metal screws. Sham operated knee joints of 66 rats had holes drilled and screws inserted but none of them were plated. The capsule from the anterior and posterior portion of the knee joints was harvested at 3 days, 1, 2, 4, 8 and 16 weeks after immobilization and the expression patterns of TGF-beta1 and CTGF were characterized using in situ hybridization and immunohistochemistry. RESULTS: The in situ hybridization demonstrated that the mRNAs of both TGF-beta1 and CTGF increased continuously during the first 2 weeks after immobilization and then decreased. The response was relatively higher in the posterior capsule than in the anterior one. In contrast, the immunoreactivity of both TGF-beta1 and CTGF increased gradually with time. The response was much stronger in the posterior capsule than in the anterior one. CONCLUSIONS: The capsule has a potency to produce TGF-beta1 and CTGF after immobilization. CTGF may play a role in causing and maintaining capsular fibrosis in collaboration with TGF-beta1. The fibrotic change in the posterior capsule may have resulted in limited motion in extension in this immobilized knee model in rats. It may be possible to prevent joint contractures by somehow blocking the fibrotic process. PMID- 18509818 TI - Comparison of placental PTEN and beta1 integrin expression in early spontaneous abortion, early and late normal pregnancy. AB - BACKGROUND: PTEN seems to play an important role in cell cycle, growth, migration, and death. Integrins are cell surface receptors that play a role in the regulation of cell proliferation, differentiation, implantation, and embryogenesis. PTEN inhibits beta1 integrin signaling. The objective of this study is to investigate the expression of PTEN and beta1 integrin in placental tissues of early spontaneous abortion and first and third trimesters of normal pregnancy. METHOD: A total of 43 placental tissue samples were evaluated using immunohistochemistry for PTEN and beta1 integrin. Group 1 included placental tissues of volunteer termination of normal pregnancy during the first trimester (5-10 wk gestation). Group 2 included placental tissues of normal vaginal delivery at the third trimester of pregnancy (36-40 wk gestation). Group 3 included placental tissues of pregnancy termination because of spontaneous abortion during the first trimester (5-10 wk gestation). RESULTS: PTEN expression of villous trophoblast was decreasing as the pregnancy advanced. PTEN staining of decidual cells was significantly stronger in tissue samples from early spontaneous abortion than in tissue samples from early and late normal pregnancy (p=0.003, p=0.001, respectively). There was no significant difference between beta1 integrin expression of villous trophoblast and decidual cells in three groups. CONCLUSION: Our findings suggest that altered patterns of PTEN expression may be associated with abortion, but it seems that beta1 integrin does not contribute to this process as a signaling protein. Further evaluation is needed to highlight this subject. PMID- 18509819 TI - [Neuropharmacology]. PMID- 18509820 TI - [Coauthorship networks and institutional collaboration in Revista de Neurologia]. AB - INTRODUCTION: Scientific cooperation is essential for the advance of science. Bibliometrics and social network analysis offer evaluation indicators to analyse collaboration in scientific papers. The aim of this study is to characterize scientific collaboration patterns in Revista de Neurologia between 2002 and 2006. MATERIALS AND METHODS: Coauthorships and institutional relationships of papers published in Revista de Neurologia have been identified. Collaboration Index, the most productive authors' and institutional collaboration patterns and the types of institutional collaborations have been quantified. Also, it has been constructed the coauthorship networks and the institutional collaboration network. Networks have been identified and represented using Access and Pajek software tools. RESULTS: The Collaboration Index was 4.01. 56.54% of papers involved institutional collaboration. The collaboration between institutions of the same country prevails (52.7%), followed by collaborations between departments, services or units of the same institution (40.47%) and international collaboration (6.83%). 45 coauthorship networks involving 149 investigators with a high intensity of collaboration and a large institutional network involved 80 centres were observed. CONCLUSIONS: Revista de Neurologia covers scientific production of a high number of research groups. It has been observed a positive evolution in the collaboration patterns over the time. Nevertheless, it is essential to encourage inter-regional and international collaboration. PMID- 18509821 TI - [Study on the quality of life in patients with ischaemic stroke]. AB - AIM: To study the quality of life (QoL) and functional neurological status of patients three years after suffering their first ischaemic cerebrovascular accident (CVA). PATIENTS AND METHODS: For the study we chose patients who presented ischaemic CVA, classified in categories I 63 to I 69. QoL and functional neurological status were measured using the modified Rankin Scale (mRS) and the SF-36 health questionnaire, respectively. RESULTS: Of a total of 59 patients who answered the surveys (mean age: 62 years; sex: 51% females), nearly half of them (29 patients; 49.1%) presented some kind of disability (mRS = 2). Thirty patients (50.9%) were independent or presented minimum sequelae (mRS = 1) at the end of the follow-up period. The mean QoL with the SF-36 questionnaire progressively diminished as the mRS score increased, especially in the group with mRS = 3. Although the youngest age group (range: 20-36 years) got the best average score (84 points) on the SF-36, no significant differences were found with the remaining age groups; the group made up of 75-year-olds and above was the one that obtained the lowest average score (63 points). Extensive strokes (total anterior circulation infarctions) of a cardioembolic origin (atrial fibrillation) had the lowest QoL indexes (p < 0.05). CONCLUSIONS: In the population that was analysed, the QoL was inversely proportional to the age and the severity of the functional deficit of patients who survived a completed stroke, especially in the cardioembolic-origin subgroup, due to its being associated with more extensive cerebral infarcts. PMID- 18509822 TI - [Chronic inflammatory demyelinating polyradiculoneuropathy. 10 years' experience in a Mexican centre]. AB - INTRODUCTION: Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is a chronic, but potentially treatable, acquired autoimmune neuropathy. A review of the literature shows that few studies have been conducted on its epidemiology, presenting symptoms and long-term functional prognosis. AIM: To describe the clinical and neurophysiological forms of patients with CIDP at the outset and their follow-up at one year. PATIENTS AND METHODS: We conducted a descriptive, retrospective study of patients who were hospitalised in our unit between 1995 and 2005. The cases were defined in accordance with Inflammatory Neuropathy Cause and Treatment (INCAT) group criteria. Data gathered included demographic characteristics, forms of clinical presentation, neurophysiological findings, cerebrospinal fluid and functional prognosis at one year. A statistical descriptive analysis was performed. RESULTS: The sample consisted of 26 patients- 12 males (46.15%) and 14 females (53.84%)--between 15 and 71 years of age (40.17 +/- 15.7 years). CIDP was associated with other autoimmune diseases in 20.8% of the patients. The predominant features at the outset of the disease were paresis and distal symmetrical paresthesias in the four limbs, high protein levels in cerebrospinal fluid and demyelination with axonal degeneration. Prednisone was administered in 43% of the cases. At one year, five patients remained asymptomatic (22.72%), there was a partial improvement in 13 (59.09%) and no improvement was seen in four cases (18.18%). CONCLUSIONS: The most frequent initial form of clinical presentation of CIDP in our population is quadriparesis and distal symmetrical paresthesias, high protein levels in cerebrospinal fluid and demyelination with axonal degeneration, which are related to a good functional prognosis at one year. PMID- 18509823 TI - [Epileptiform activity in psychiatric disorders in children]. AB - INTRODUCTION: Abnormalities are often observed in the electrical activity of the brain in patients with brain dysfunction. PATIENTS AND METHODS: We conducted a retrospective analysis of the results of electroencephalogram studies carried out between August and December 2006 on children who had been clinically diagnosed with a psychiatric disorder, without taking into account those with epilepsy. RESULTS: A total of 1000 electroencephalogram (EEG) studies were reviewed; the age of the patients ranged from 0 to 18 years, with an average of 9.7 years, and the ratio between sexes was 67.9% males and 32.1% females. The four most frequently observed psychiatric disorders were: attention deficit hyperactivity disorder, 422; depressive disorder, 103; mental retardation, 99; and behavioural disorder, 96. The EEG findings were divided into normal (37.8%) and abnormal patterns (61.9%). The most frequent abnormality was bioelectric immaturity (56.4% of cases). On analysing epileptiform activities, we found 26 studies (2.6%) and evidence of cortico-subcortical irritability in 6.7%. The total number of children with epileptiform patterns was 71. The most common pattern was the sharp wave (97.2%) and the most frequent location was the temporal lobe (42.3%). CONCLUSIONS: A high percentage of patients with psychiatric disorders also have abnormalities in the electrical activity of their brains, and even epileptiform activity. Electrical abnormalities in the general population of healthy children have been reported in the literature, but the figures that we found are higher for patients with psychiatric disorders. Functioning of the neurons is incorrect and this results in a range of different pathologies. The repercussion of abnormal electrical activity of the brain on the clinical data of the patients is open to discussion, and we consider it to be an epiphenomenon. PMID- 18509824 TI - [Compromise of the cognitive functions in progressive secondary multiple sclerosis]. AB - AIM: To define the patterns of cognitive impairment in a homogeneous group of secondary progressive multiple sclerosis (SPMS) patients. PATIENTS AND METHODS: Forty-two SPMS patients were included with a similar degree of disability; all had been treated with interferon beta-1b for a minimum of 3 months. They voluntarily complimented a battery of 10 neuropsychological tests selected for this study, distributed in two sessions of one hour. In addition, the emotional state was evaluated with the Beck Depression Inventory and the Hamilton Anxiety Scale. We considered cognitive impairment as more than two tests altered, according with previously reported studies. RESULTS: 73.8% of patients were women; mean age was 45 years (range: 25-62); mean EDSS was 5.4 (range: 3.0-7.5); mean evolution time was 34.5 months (range: 24-80); mean treatment duration was 13.5 months (range: 3-38). Cognitive impairment was present in 78.5% of patients. The most frequently impaired functions were: attentional capacity, visuospatial perception, verbal fluency, short-term and long-term logic memory and abstract reasoning. The presence of cognitive impairment was related to the time of evolution of the disease (r = 0.31; p < 0.05) but not with the age, the degree of disability or the treatment duration. CONCLUSION: Cognitive impairment in the SPMS patients is a frequent finding, being the alteration in the speed for the acquisition and processing of new information, and the abstract reasoning the most frequent and severe altered functions. The also frequent impairment of visuospatial information was a differential finding in our study that could contribute to diagnosis of clinical progression. PMID- 18509825 TI - [Lymphomatosis cerebri as the cause of leukoencephalopathy]. AB - INTRODUCTION: Lymphomatosis cerebri (LC) is an infrequent type of primary lymphoma of the central nervous system that is characterised by diffuse, infiltrating involvement of the white matter of the brain without the formation of a mass. AIM: To report the case of a patient with LC in order to draw attention to this disease, which is rarely diagnosed, and to its initial presentation in magnetic resonance imaging (MRI) as leukoencephalopathy. CASE REPORT: Our patient was a 56-year-old female who had clinical signs and symptoms of sub-acute dementia. Computerised axial tomography and MRI of the head revealed extensive, diffuse and bilateral involvement of the white matter, basal nuclei, mesencephalon and pons, with no mass effect or contrast enhancement. A stereotactic biopsy of the white matter (which was not conclusive) showed a perivascular mixed mononuclear-cell inflammatory infiltrate of B and T cells. No cytologic atypia was observed. Treatment was established with corticoids, which produced a clinical and radiological improvement in the first two months. During the next month the patient underwent rapid clinical deterioration with sleepiness and a worsening of the ability to walk. In an MRI scan the lesion had a more heterogeneous appearance with mass effect on adjacent structures and patchy contrast enhancement. A wedge biopsy of brain tissue led to a diagnosis of high grade B-cell lymphoma. CONCLUSIONS: The imaging and histological appearance of LC may not be the one typically found in primary lymphomas of the central nervous system, and its clinical presentation may be similar to that of other diffuse processes involving compromise of the white matter (cerebral gliomatosis, inflammatory diseases of the white matter, such as Behcet's disease, Sjogren's disease or systemic lupus erythematosus). PMID- 18509826 TI - [Ivermectin as a therapeutic alternative in neurocysticercosis that is resistant to conventional pharmacological treatment]. AB - INTRODUCTION: Neurocysticercosis is a public health problem that can be found in many parts of the world, especially in developing countries, and today's high rates of immigration are making it increasingly more common in developed countries. Cysticidal treatment of neurocysticercosis is a controversial issue because it is only partially effective against vesicular and colloidal-shaped cysts when the parasite persists after a course of albendazole or praziquantel, the only two therapeutic options that are currently available. Ivermectin is a very effective, safe veterinary and human antiparasitic drug, with occasional very mild side effects. It has been used for over 25 years in cases of endoparasitosis that do not respond well to treatment, such as filariasis, oncocerciasis, strongyloidiasis, etc. and also in ectoparasitoses, such as pediculolsis capitis and myasis. It acts in the myoneural junction on the receptors in the chloride channel by increasing their permeability and causing paralysis in adult worms or by a mediated immune mechanism when it acts on immature forms. CASE REPORTS: We report the cases of four patients who were previously treated with albendazole reiteratively with radiological evidence, which shows the persistence of viable vesicular or colloidal-shaped cysts. These patients were given 10 mg/day of ivermectin for 15 consecutive days or 10 mg/day as an average for 30 days, with excellent clinical and radiological progress. CONCLUSIONS: Ivermectin was effective and did not give rise to any side effects when used to treat these four patients, who were resistant to conventional treatment with albendazole and/or praziquantel. PMID- 18509827 TI - [Inflammatory/anti-inflammatory mechanisms in the brain following exposure to stress]. AB - INTRODUCTION: Most of the biological systems that go to make up an organism can be affected by stress. The central nervous system not only plays an essential role in regulating the general response to stress, but it is also one of its main targets. The consequences may be positive (for example, a state of alertness) or negative (neuropsychiatric pathologies). More specifically, exposure to certain stressing stimuli can trigger a neuroinflammatory process. DEVELOPMENT: Reports have appeared describing how an excessive neuroinflammatory response makes a decisive contribution to the functional and structural damage that is often observed in stress-related neurological and neuropsychiatric diseases, such as post-traumatic stress syndrome, depression and schizophrenia. The inflammatory process generated by exposure to stress is characterised by a complex release of a chain of different cell mediators, such as cytosines, transcription factors, prostaglandins, free radicals, and so forth. In parallel to this, it has been proved that the anti-inflammatory pathway of deoxyprostaglandins is activated after stress in the central nervous system, and this activation could constitute an endogenous mechanism that regulates the inflammatory process itself. CONCLUSIONS: In the future, further studies and a deeper understanding of this endogenous pathway could make it into a new, interesting preventive or neuroprotective strategy for use in a number of pathologies that have a clear harmful inflammatory component, such as cerebral ischaemia, Alzheimer's and Parkinson's diseases, as well as those mentioned earlier as being related to exposure to stress. PMID- 18509828 TI - [Models of executive control and functions (I)]. AB - INTRODUCTION: As human beings we are capable of coping with novel situations and adapting to changes in a flexible manner. The cognitive skills that allow individuals to control and regulate their behaviour are called executive functions. Anatomically, the executive functions depend on a distributed neural system, in which the prefrontal cortex plays an essential role. Recent data suggest that different regions of the prefrontal cortex may be involved in a number of aspects of executive functioning. DEVELOPMENT: The purpose of this article is to carry out a review of the main models of executive functioning in order to shed light on this controversial construct. The models put forward to date approach the same reality from a number of different perspectives, in some case avoiding certain parts of that reality. In this first part, we review the models and theories of contextual information, structured complex events, working memory, adaptive encoding, Miller and Cohen's integrating theory, and the factorial models of executive control. CONCLUSIONS: There is no single model that allows us to explain how specific cognitive processes are controlled and coordinated while complex cognitive activities are being performed. Nevertheless, some agreement has been reached on accepting the idea that the executive functions construct does not consist of a single concept, but instead a combination of several cognitive processes that combine in a number of ways in order to operate in different situations. PMID- 18509829 TI - [Paraneoplastic striatal encephalitis associated with anti-CV2]. PMID- 18509830 TI - [XII Reunion de la Sociedad Canaria de Neurologia. Communications]. PMID- 18509831 TI - [Small vessel vasculitis associated with the use of interferon beta-1a in multiple sclerosis]. PMID- 18509832 TI - [Leprosy, its diagnosis by the naked eye]. PMID- 18509833 TI - [Pseudohypertrophic myopathy as a manifestation of hypothyroidism (Kocher-Debre Semelaigne syndrome)]. PMID- 18509834 TI - [Segmental spinal dysgenesis]. PMID- 18509835 TI - Diffusion of interferon beta in Iran and its utilization in Tehran. AB - PURPOSE: There are few studies on the diffusion of expensive new therapies in low or middle-income countries. The objectives of this study were to describe the diffusion of interferon beta for the treatment of patients with multiple sclerosis (MS) in Iran and to analyze its use in a cohort of 890 patients in Tehran registered in the Iranian MS Society (IMSS) registry. METHODS: Data on the diffusion of interferon beta drugs from 2000 to 2004 in Iran were obtained from the Ministry of Health (MOH). Data on the utilization of interferon beta in selected patient groups were collected from routine clinical data of the IMSS in a sample of population in Tehran province from 2003 to 2005. RESULTS: Interferon beta had a rapid diffusion in Iran during the study period, and it was used in all 28 provinces in 2004, however, much less in the poor provinces. Interestingly, interferon beta was prescribed to 68% of the patients in Tehran registered in the IMSS registry during 2003-2005. The usage of interferon beta as registered in the IMSS was higher (p < 0.0001) among insured than uninsured patients. CONCLUSION: The use of interferon beta treatment has grown considerably in Iran. In Tehran, interferon beta seems to be mainly used by young ambulatory patients with relapsing-remitting MS, indicating a rational drug use. PMID- 18509836 TI - A valence bond study of the low-lying states of the NF molecule. AB - The electronic structures of the three lowest-lying states of NF are investigated by means of modern valence bond (VB) methods such as the VB self-consistent field (VBSCF), breathing orbital VB (BOVB), and VB configuration interaction (VBCI) methods. The wave functions for the three states are expressed in terms of 9-12 VB structures, which can be further condensed into three or four classical Lewis structures, whose weights are quantitatively estimated. Despite the compactness of the wave functions, the BOVB and VBCI methods reproduce the spectroscopic properties and dipole moments of the three states well, in good agreement with previous computational studies and experimental values. By analogy to the isoelectronic O(2) molecule, the ground state (3)Sigma(-) possesses both a sigma bond and 3-electron pi bonds. However, here the polar sigma bond contributes the most to the overall bonding. It is augmented by a fractional (19%) contribution of three-electron pi bonding that arises from pi charge transfer from fluorine to nitrogen. In the singlet (1)Delta and (1)Sigma(+) excited states the pi-bonding component is classically covalent, and it contributes 28% and 37% to the overall bonding picture for the two states, respectively. The resonance energies are calculated and reveal that pi bonding contributes at least 24, 35 and 42 kcal mol(-1) to the total bonding energies of the (3)Sigma(-), (1)Delta and (1)Sigma(+) states, respectively. Some unusual properties of the NF molecule, like the equilibrium distance shortening and bonding energy increasing upon excitation, the counterintuitive values of the dipole moments and the reversal of the dipole moments as the bond is stretched, are interpreted in the light of the simple valence bond picture. The overall polarity of the molecule is very small in the ground state, and is opposite to the relative electronegativity of N vs F in the singlet excited states. The values of the dipole moments in the three states are quantitatively accounted for by the calculated weights of the VB structures. PMID- 18509837 TI - Ionic liquids and proteases: a clean alliance for semisynthesis. AB - Herein we present the first report on protease-catalysed ligation of cleavage sensitive peptide and protein fragments in ionic-liquid-containing solvent systems. By applying the newly established [MMIM][Me2PO4]/buffer mixture as a reaction medium, significant advantages over purely aqueous or conventional organic solvent-containing media could be identified, including in particular the use of active wild-type proteases as biocatalysts, the suppression of any competitive proteolytic side reactions, the high turnover rates compared to classical organic solvents and the high stability of chemically labile reactants. PMID- 18509838 TI - Calix[n]arene-based glycoclusters: bioactivity of thiourea-linked galactose/lactose moieties as inhibitors of binding of medically relevant lectins to a glycoprotein and cell-surface glycoconjugates and selectivity among human adhesion/growth-regulatory galectins. AB - Growing insights into the functionality of lectin-carbohydrate interactions are identifying attractive new targets for drug design. As glycan recognition is regulated by the structure of the sugar epitope and also by topological aspects of its presentation, a suitable arrangement of ligands in synthetic glycoclusters has the potential to enhance their avidity and selectivity. If adequately realized, such compounds might find medical applications. This is why we focused on lectins of clinical interest, acting either as a potent biohazard (a toxin from Viscum album L. akin to ricin) or as a factor in tumor progression (human galectins-1, -3, and -4). Using a set of 14 calix[n]arenes (n=4, 6, and 8) with thiourea-linked galactose or lactose moieties, we first ascertained the lectin binding properties of the derivatized sugar head groups conjugated to the synthetic macrocycles. Despite their high degree of flexibility, the calix[6,8]arenes proved especially effective for the plant AB-toxin, in the solid phase model system with a single glycoprotein (asialofetuin) and with human tumor cells in vitro. The bioactivity of the calix[n]arenes was also proven for human galectins. Notably, selectivity for the tested tandem-repeat-type galectin-4 among the three subgroups was determined at the level of solid-phase and cell assays, the large flexible macrocycles again figuring prominently as inhibitors. Alternate and cone versions of calix[4]arene with lactose units distinguished between galectins-1 and -4 versus galectin-3 in cell assays. The results thus revealed bioactivity of galactose-/lactose-presenting calix[n]arenes for medically relevant lectins and selectivity within the family of adhesion/growth regulatory human galectins. PMID- 18509839 TI - Pt(II)-catalyzed synthesis of 9-oxabicyclo[3.3.1]nona-2,6-dienes from 2-alkynyl-1 carbonylbenzenes and allylsilanes by an allylation/annulation cascade. PMID- 18509840 TI - Supramolecular tandem enzyme assays for multiparameter sensor arrays and enantiomeric excess determination of amino acids. AB - The coupling of an enzymatic transformation with dynamic host-guest exchange allows the unselective binding of macrocycles to be used for highly selective analyte sensing. The resulting supramolecular tandem enzyme assays require the enzymatic substrate and its corresponding product to differ significantly in their affinity for macrocycles, for example, cation receptors, and to show a differential propensity to displace a fluorescent dye from its host-guest complex. The enzymatic transformation results in a concomitant dye displacement that can be accurately followed by optical spectroscopy, specifically fluorescence. By exploiting this label-free continuous enzyme assay principle with the fluorescent dye Dapoxyl and the macrocyclic host cucurbit[7]uril, a multiparameter sensor array has been designed, which is capable of detecting the presence of amino acids (e.g. histidine, arginine, lysine, and tyrosine) and their decarboxylases. Only in the presence of both, the particular amino acid and the corresponding decarboxylase, is the amine or diamine product formed. These products are more highly positively charged than the substrate, have a higher affinity for the macrocycle and, therefore, displace the dye from the complex. The extension of the high selectivity and muM sensitivity of the tandem assay principle has also allowed for the accurate measurement of D-lysine enantiomeric excesses of up to 99.98 %, as only the L-enantiomer is accepted by the enzyme as a substrate and is converted to the product that is responsible for the observed fluorescence signal. PMID- 18509841 TI - Molecular, crystal, and thin-film structures of octathio[8]circulene: release of antiaromatic molecular distortion and lamellar structure of self-assembling thin films. PMID- 18509842 TI - The tandem ring-closing metathesis-isomerization approach to 6-deoxyglycals. AB - Protected 3,6-dideoxyglycals have been synthesized de novo as single isomers starting from ethyl lactate by using the tandem RCM-isomerization reaction as the key step. Different relative configurations become accessible by addition of vinyl- or allyl-metal compounds to protected lactaldehydes under Cram-chelate or Felkin-Anh control. The concept is exemplified for glycals of L-rhodinose and L amicetose, as well as for ring-expanded non-natural analogues thereof. This novel approach to glycals is also applicable to the synthesis of disaccharide glycals via a reiterative strategy, as exemplified for the dimer of L-rhodinose and its non-natural ring expanded analogue. PMID- 18509843 TI - Biocompatible carbon nanotubes generated by functionalization with glycodendrimers. PMID- 18509844 TI - Colour change of chemiluminescence for base-induced decomposition of dioxetane bearing a 4-(4-cyanophenyl)iminomethyl-3-hydroxyphenyl group. AB - A bicyclic dioxetane 1 bearing a 4-(4-cyanophenyl)iminomethyl-3-hydroxyphenyl group was found to undergo base-induced decomposition with the accompanying emission of light, the colour of which changed depending on the base used and its concentration. When 1 was triggered with tetrabutylammonium fluoride (TBAF), 1 displayed an emission of glowing orange light. On the other hand, on treatment with a high concentration of potassium t-butoxide complexed with 18-crown-6 ether, 1 afforded a flash of blue light. The mechanistic study of this unprecedented phenomenon revealed that the emission of glowing orange light was due to the normal oxido anion of keto ester 11, whereas the emission of a flash of blue light was attributed to another species that was produced by addition of a nucleophile to an iminomethyl of unstable oxido anion of dioxetane 10. PMID- 18509845 TI - The best approach for early prediction of fetal gender by using free fetal DNA from maternal plasma. AB - OBJECTIVES: Detection of free fetal DNA (ffDNA) in maternal blood during pregnancy has given rise to the possibility of developing new noninvasive approaches for early prenatal diagnosis. On a large-scale study, two protocols of real-time polymerase chain reaction (PCR) were compared in order to establish which Y-specific locus, either multicopy DYS14 or single copy SRY sequence, was the most suitable for developing a test with high diagnostic efficiency for early fetal gender assessment. The second aim was to assess whether the combination of the two detection systems could increase the performance of the prenatal test. METHODS: We analyzed 145 plasma samples from healthy pregnant women between 11 and 12 weeks of singleton gestation. For each sample, fetal gender was determined by using both protocols (DYS14 and SRY) during the same real-time PCR run. RESULTS: The data obtained by the DYS14 and SRY assays showed an efficiency in fetal gender prediction of 97.9 and 80%, respectively. It is not advisable to combine the two protocols because this association does not help in further improvements in fetal gender prediction. CONCLUSIONS: DYS14 assay is the best approach for early fetal gender assessment because it is more sensitive, accurate, and efficient than the SRY assay. PMID- 18509846 TI - Recent allopolyploid origin of Zygosaccharomyces rouxii strain ATCC 42981. AB - Zygosaccharomyces rouxii strain ATCC 42981 has been reported to have two copies of several genes including HOG1 and SOD2, whereas the type strain of Z. rouxii (CBS 732) has only one. To investigate the structure of the ATCC 42981 genome we sequenced random fragments from this genome and compared the data to the type strain. We found that ATCC 42981 contains two versions of the ribosomal RNA array, one of which is identical in the ITS1-ITS2 and 26S D1/D2 regions to Z. rouxii CBS 732, while the other is almost identical to a species provisionally named Z. pseudorouxii. We found that most genomic regions from Z. rouxii CBS 732 map in a one-to-two fashion to pairs of regions in ATCC 42981, with one of the ATCC 42981 regions having 97-100% DNA sequence identity to CBS 732 and the other having about 80-90% identity. Complete sequencing of regions containing 30 pairs of genes from ATCC 42981 and their orthologues in CBS 732 showed no evidence of the gene deletions or pseudogene formation that might be expected if ATCC 42981 had undergone whole-genome duplication several million years ago and was in the early stages of gene loss. Instead, we conclude that ATCC 42981 is a Z. rouxii-Z. pseudorouxii interspecies hybrid that was formed so recently that its genome has not had time to decay. PMID- 18509847 TI - Common industrial sake yeast strains have three copies of the AQY1-ARR3 region of chromosome XVI in their genomes. AB - Genomic analysis of industrial yeast strains is important for exploitation of their potential. We analysed the genomic structure of the most widely used sake yeast strain, Kyokai no. 7 (K7), by DNA microarray. Since the analysis suggested that the copy number of the AQY1-ARR3 region in the right arm of chromosome XVI was amplified, we performed Southern blot analysis using the AQY1 gene as a probe. The probe hybridized to three bands in the widely used sake strains derived from K7, but only to one band of 1.4 kb in the laboratory strains. Since the extra two bands were not observed in old sake strains, or in other industrial strains, the amplification of this region appeared to be specific for the widely used sake strains. The copy number of the AQY1-ARR3 region appeared to have increased by chromosomal translocation, since chromosomal Southern blot analysis revealed that the AQY1 probe hybridized to chromosomes IV and XIII, in addition to chromosome XVI, in which AQY1 of the laboratory strain is encoded. The chromosomal translocation was also confirmed by PCR analysis using primers that amplify the region containing the breakpoint. Cloning and sequencing of cosmids that encode the AQY1-ARR3 region revealed that this region is flanked by TG(1-3) repeats on the centromere-proximal side in chromosomes IV and XIII, suggesting that amplification of this region occurred by homologous recombination through TG(1-3) repeats. These results demonstrate the genomic characteristics of the modern widely used sake strains that discriminate them from other strains. PMID- 18509848 TI - A history of research on yeasts 12: medical yeasts part 1, Candida albicans. PMID- 18509849 TI - Chromosome instability and unusual features of some widely used strains of Candida albicans. AB - Electrophoretic karyotyping of the Candida albicans revealed a different migration pattern of ChR in three different stocks of the sequencing strain SC5314. In one stock, the high instability of ChR size prevented the migration of ChR as a compact band; ChR appeared, instead, as a smear. In some stocks, ChR and/or Ch1 ploidy diminished, suggesting mixed populations of disomic and monosomic cells. Similarly, some stocks of widely used derivatives CAI4 and BWP17 contained smearing of ChR. In addition, the most manipulated strain in the lineage of SC5314, the last derivative, BWP17, acquired an increase in the size of Ch7b and revealed an unusual property. BWP17 did not tolerate a well established procedure of telomere-mediated fragmentation of a chromosome; the remaining intact homologue always duplicated. We suggest that some stocks of SC5314 are unstable and that BWP17 may not be appropriate for general studies. Instead of BWP17 or CAI4, we recommend using for general research CAF4-2, which is a relatively stable Ura- derivative, and which has been successfully used for more than a decade in our laboratory. PMID- 18509851 TI - Antidepressant treatments regulate matrix metalloproteinases-2 and -9 (MMP-2/MMP 9) and tissue inhibitors of the metalloproteinases (TIMPS 1-4) in the adult rat hippocampus. AB - Antidepressants induce structural remodeling in the adult hippocampus, including changes in dendritic arbors, axonal sprouting, neurogenesis, and endothelial cell proliferation. Such forms of structural plasticity take place in the context of the extracellular matrix environment and are known to be regulated by matrix metalloproteinases (MMPs), in particular MMP-2/9, and their endogenous regulators, the tissue inhibitors of the metalloproteinases (TIMPs 1-4). Given the hippocampal structural remodeling associated with antidepressant treatments, we hypothesized that antidepressants may regulate the expression and activity of MMP-2/9 and TIMPs 1-4. The influence of distinct classes of antidepressants, namely, electroconvulsive seizure, fluoxetine, tranylcypromine, and desipramine, on the gene expression of MMP-2, MMP-9, and TIMPs 1-4 in the hippocampus was determined using radioactive in situ hybridization. In addition, zymography studies addressed the regulation of the gelatinase activity of MMP-2/9 following acute and chronic antidepressant administration. We observed that acute and chronic ECS differentially regulate the transcript levels of MMP-2/9 and TIMPs 1 4 and also increase gelatinase activity in the hippocampus. Acute and chronic pharmacological antidepressants on the other hand differentially alter the expression of the TIMPs without any observed effect on hippocampal MMP-2/9 expression or activity. These findings raise the possibility that extracellular matrix modifying enzymes and their endogenous regulators may serve as targets for antidepressant treatments and suggests the possibility that they may contribute to antidepressant-mediated structural plasticity in the hippocampus. PMID- 18509852 TI - Differential synaptic changes in the striatum of subjects with undifferentiated versus paranoid schizophrenia. AB - Subjects with schizophrenia (SZ) have an increased density of synapses characteristic of corticostriatal or thalamostriatal glutamatergic inputs in the caudate matrix and putamen patches. SZ is a heterogeneous disease in many aspects including symptoms. The purpose of the present study was to determine if the synaptic organization in two different DSM-i.v. subgroups of SZ was differentially affected. Postmortem striatal tissue was obtained from the Maryland Brain Collection from normal controls (NC), chronic paranoid SZs (SZP), and chronic undifferentiated SZs (SZU). Tissue was prepared for calbindin immunocytochemistry to identify patch matrix compartments, prepared for electron microscopy and analyzed using stereological methods. The synaptic density of asymmetric synapses, characteristic of glutamatergic inputs, was elevated equivalently in striatal patches in the SZP and SZU versus NC. The SZU also had an increased density of asymmetric synapses in the striatal matrix compared to NC. Moreover, symmetric axospinous synapses, characteristic of intrinsic inhibitory inputs and dopaminergic afferents, showed a dichotomy in synaptic density between the SZU and SZP in the striatal and caudate matrix. These data show discreet differences in synaptic organization between SZU and SZP and/or NCs. The results suggest that abnormal corticostriatal and/or corticothalamic inputs to striatal patches may be related to limbic dysfunction, which is perturbed in both subtypes of SZ. The selective increase in axospinous synapses in the matrix of the SZU subgroup compared to the SZP may be related to more severe cognitive problems in that subset of SZ compared to SZP. PMID- 18509853 TI - Microscopic computed tomography imaging of the cerebral circulation in mice: feasibility and pitfalls. AB - This study evaluates the utility and practical limitations of microcomputerized X ray tomography (CT) as a research tool for examination of the cerebral circulation in mice. Six micro CT angiograms of the circle of Willis (COW) from six mice were obtained by scanning whole head and brain specimen perfused with a radio-opaque silicone contrast agent. Two-dimensional volume rendered images were postprocessed from three-dimensional image datasets using a partially automated high-throughput model that generated 10 surface projections for each specimen. The image processing model employed a straightforward global thresholding and computerized component labeling software algorithm. Postprocessed images were analyzed and results correlated with microdissection. Micro CT demonstration of COW vessels and their branch anatomy was assessed. 71% of COW vessels were completely demonstrated, 26% were partially demonstrated, and 3% were not demonstrated. All cases of nondemonstration and most cases of partial demonstration resulted from scan coverage or postprocessing clip error. Thresholding effect caused pseudostenosis of 8% of COW vessels and accounted for a minority of partial demonstration cases. No imaging artifacts were caused by contrast extravasation or ineffective contrast perfusion. Volume averaging caused minor angioarchitectural distortion of 58% of COW vessels. Ninety-five percent of COW > or =50 microm and 52% of COW vessels <50 microm were correctly identified by micro CT. Micro CT of the murine COW using a high-throughput image processing model is feasible. Angioarchitectural distortion due to volume averaging and thresholding effect can occur and pathological findings should be confirmed. PMID- 18509854 TI - Genetic and pharmacological manipulations of the CB(1) receptor alter ethanol preference and dependence in ethanol preferring and nonpreferring mice. AB - Recent studies have indicated a role for the endocannabinoid system in ethanol related behaviors. This study examined the effect of pharmacological activation, blockade, and genetic deletion of the CB(1) receptors on ethanol-drinking behavior in ethanol preferring C57BL/6J (B6) and ethanol nonpreferring DBA/2J (D2) mice. The deletion of CB(1) receptor significantly reduced the ethanol preference. Although the stimulation of the CB(1) receptor by CP-55,940 markedly increased the ethanol preference, this effect was found to be greater in B6 than in D2 mice. The antagonism of CB(1) receptor function by SR141716A led to a significant reduction in voluntary ethanol preference in B6 than D2 mice. A significant lower hypothermic and greater sedative response to acute ethanol administration was observed in both the strains of CB(1) -/- mice than wild-type mice. Interestingly, genetic deletion and pharmacological blockade of the CB(1) receptor produced a marked reduction in severity of handling-induced convulsion in both the strains. The radioligand binding studies revealed significantly higher levels of CB(1) receptor-stimulated G-protein activation in the striatum of B6 compared to D2 mice. Innate differences in the CB(1) receptor function might be one of the contributing factors for higher ethanol drinking behavior. The antagonists of the CB(1) receptor may have therapeutic potential in the treatment of ethanol dependence. PMID- 18509855 TI - Reuptake of L-DOPA-derived extracellular DA in the striatum of a rodent model of Parkinson's disease via norepinephrine transporter. AB - To determine the role of norepinephrine transporter in reuptake of L-DOPA-derived extracellular DA in the DA-denervated Parkinsonian striatum, we examined extracellular DA levels in the striatum of 6-hydroxyDA-lesioned rats that received L-DOPA (50 mg/kg with 12.5 mg/kg of benserazide) and L-DOPA plus desipramine (25 mg/kg), a selective norepinephrine reuptake inhibitor, using in vivo microdialysis. The pretreatment with desipramine increased levels of extracellular DA derived from administrated L-DOPA in the DA-denervated striatum. This study provides evidence that L-DOPA-derived DA is taken up by the norepinephrine transporter, instead of the dopamine transporter, in the striatum with dopaminergic denervation. This result suggests that the norepinephrine transporter could be a promising target in the treatment for Parkinson's disease. PMID- 18509856 TI - Congenital diaphragmatic hernia in a first-trimester ultrasound aneuploidy screening program. AB - OBJECTIVES: To review our experience with the prenatal detection of congenital diaphragmatic hernia (CDH) in fetuses presenting for ultrasound screening of chromosomal abnormalities in the first trimester. METHODS: As part of our first trimester ultrasound protocol, fetuses with a crown-rump length between 45 and 84 mm underwent a limited anatomical assessment in conjunction with nuchal translucency thickness measurement and nasal bone assessment. Cases of CDH diagnosed prenatally or after delivery in this population were identified. RESULTS: Among the six cases of CDH detected (prevalence of 1 in 927), the first trimester ultrasound findings were abnormal in five fetuses (83%), including three with increased nuchal translucency only; one with increased nuchal translucency, an intrathoracic stomach, dextrocardia and a cephalocele; and one with normal nuchal translucency thickness and a small, complex intrathoracic mass later confirmed as the fetal stomach. The diagnosis of CDH was confidently made in the first trimester in one case, in the second trimester in three cases, and after birth in the remaining two cases. CONCLUSION: The diagnosis of CDH in the first trimester is difficult, especially in those cases in which the defect is small or late migration of the abdominal viscera occurs. Therefore, screening for CDH in the first trimester is unlikely to be effective. PMID- 18509857 TI - Interstitial laser therapy for fetal reduction in monochorionic multiple pregnancy: loss rate and association with aplasia cutis congenita. AB - OBJECTIVE: To evaluate experience with interstitial laser therapy for intrafetal vascular ablation in monochorionic (MC) multiple pregnancy. METHODS: MC pregnancies that underwent fetal reduction between 1998 and 2007 by interstitial laser therapy were reviewed. Indications were twin reversed arterial perfusion sequence (TRAP) (n = 10), twin-to-twin transfusion (6), discordant abnormality (7) or growth (1) and high-order multiples (6). RESULTS: Thirty pregnancies treated at 15 weeks (median, range: 11 weeks-20 weeks, 5 days) had no technical failures but four manifested procedure-related amniorrhexis. Four of 38 remaining fetuses suffered intrauterine death (IUFD) within 24 h, giving an early procedure related fetal loss rate of 10% per pregnancy and 11% per fetus. A further five IUFDs occurred within 2 weeks, giving a maximum procedure-related loss rate of 27% per pregnancy and 24% per fetus. Median gestation at delivery was 37 weeks (18 weeks, 1 day-41 weeks, 3 days) for pregnancies continuing > 2 weeks. Perinatal survival was 26 of 38 (68%) in nonreduced fetuses. Two of 26 neonates (8%) were diagnosed with aplasia cutis congenita (ACC). CONCLUSION: Interstitial laser therapy in complicated MC pregnancies carries significant risks of unintended fetal loss and may be associated with ACC. PMID- 18509858 TI - Carrier-mediated transport of metformin across the human placenta determined by using the ex vivo perfusion of the placental cotyledon model. AB - OBJECTIVE: Metformin is a polar positively charged compound. The aim of the study was to characterize its permeability across the human placenta using the ex vivo placental perfusion model. METHODS: Selected cotyledons from term placentas were cannulated and dually perfused. Metformin (10 mg/L, 1000 mg/L) and a permeability reference, antipyrine (50 mg/L), were added to the maternal circulation. Samples from maternal and fetal compartments were analyzed for metformin and antipyrine by high performance liquid chromatography (HPLC). The permeation of metformin was also studied using the parallel artificial membrane permeation assay (PAMPA) that was designed to predict passive transcellular permeability of drugs. RESULTS: In this study, 15 complete placental perfusion experimental set-ups were performed. The mean percent transport increased as metformin concentrations were raised and it was 11 +/- 1.32 and 16.92 +/- 0.98 for 10 mg/L and 1000 mg/L, respectively. The transport rate of metformin across the placenta was asymmetric yet, an active efflux against the gradient concentration could not be observed. Using the PAMPA assay, we confirmed that metformin does not cross by passive diffusion. CONCLUSION: The data suggest that metformin permeability across the placenta is mediated by a carrier that transport cationic compounds bi-directionally, with a higher transfer rate from the fetal to the maternal side. PMID- 18509859 TI - Right ventricular noncompaction associated with long QT in a fetus with right ventricular hypertrophy and cardiac arrhythmias. PMID- 18509860 TI - Prenatal two- and three-dimensional sonographic diagnosis of dacryocystocele. PMID- 18509861 TI - Reduced adhesion formation following laparoscopic versus open colorectal surgery. AB - BACKGROUND: Adhesion formation is common after abdominal surgery. This study aimed to compare the extent of adhesion formation following laparoscopic and open colorectal surgery. METHODS: An observational study was undertaken to identify adhesions in patients undergoing laparoscopy after previous laparoscopic or open colectomy. Adhesions were scored according to a system validated for interobserver (median kappa = 0.80) and intraobserver (kappa = 0.82) agreement. The primary endpoint was the overall adhesion score (0-10); a secondary endpoint was the adhesion score at the main incision site (0-6). RESULTS: Forty-six patients were recruited (13 laparoscopic and 33 open colectomy). In most patients (n = 29), laparoscopy was performed for tumour staging before liver resection. The median (interquartile range) overall adhesion score was 7 (5-8) in the open group and 0 (0-3) in the laparoscopic group (P < 0.001). A similar difference was found for the main incision score: 6 (4-6) versus 0 (0-0) (P < 0.001). CONCLUSION: There may be a reduction in adhesion formation following laparoscopic compared with open colectomy, although the small sample size limits this conclusion. PMID- 18509862 TI - 1H and 13C NMR spectral assignments of some novel 2,4,6,8-tetraaryl-3,7 diazabicyclo[3.3.1]nonan-9-one derivatives. AB - The (1)H and (13)C NMR spectra of 2,4,6,8-tetraaryl-3,7-diazabicyclo[3.3.1]nonan 9-ones (1-2), oximes (3-8) and O-benzyl oximes (9-12) were recorded. The chemical shifts were unambiguously assigned using 1D and 2D NMR spectral data. The results clearly indicate that the compounds exist in chair-boat conformation with equatorial and axial orientation of the aryl groups in the chair and boat forms, respectively. Since the molecules are flexible and dynamic in solution, the chair and boat forms are mutually interconvertible. In 3-12, because of the effect of oximation/oximination, all the protons in the heterobicyclic systems gave distinct signals except the benzylic protons of the boat form. In all synthesized compounds, the aryl group protons at C-6,8 are shielded by the aryl groups at C 2,4 and therefore appear in the lower frequency region than the aryl groups at C 2,4. PMID- 18509863 TI - Prenatal diagnosis of a fetus with 7q11.23 deletion detected by multiplex ligation-dependent probe amplification (MLPA) screening. PMID- 18509864 TI - Twin anemia-polycythemia sequence (TAPS) without a cause. PMID- 18509865 TI - RNAi-mediated reversible opening of the blood-brain barrier. AB - BACKGROUND: The blood-brain barrier (BBB) contains tight junctions (TJs) which reduce the space between adjacent endothelial cells lining the fine capillaries of the microvasculature of the brain to form a selective and regulatable barrier. METHODS: Using a hydrodynamic approach, we delivered siRNA targeting the TJ protein claudin-5 to the endothelial cells of the BBB in mice. RESULTS: We have shown a significant decrease in claudin-5 mRNA levels 24 and 48 hours post delivery of siRNA, with levels of protein expression decreasing up to 48 hours post-injection compared to uninjected, phosphate-buffered saline (PBS)-injected and non-targeting siRNA-injected mice. We observed increased permeability at the BBB to molecules up to 742 Da, but not 4400 Da, using tracer molecule perfusion and MRI analysis. To illustrate the functional efficacy of size-selective and transient barrier opening, we have shown that enhanced delivery of the small neuropeptide thyrotropin-releasing hormone (TRH) (MW 360 Da) to the brains of mice 48 hours post-injection of siRNA targeting claudin-5 significantly modifies behavioural output. CONCLUSIONS: These data demonstrate that it is now possible to transiently and size-selectively open the BBB in mice, allowing in principle the delivery of a wide range of agents for the establishment and treatment of experimental mouse models of neurodegenerative, neuropsychiatric and malignant diseases. PMID- 18509866 TI - Identification of oocyte-selective NLRP genes in rhesus macaque monkeys (Macaca mulatta). AB - Oocyte-selective genes control multiple aspects of female gamete development and preimplantation embryogenesis. Several key oocyte-selective factors have been identified in mice recently; however, these factors are not well documented in more advanced species such as nonhuman primates. One of such oocyte-selective factors is NLRP5 (NLR family, Pyrin domain containing 5), also known as Maternal Antigen That Embryos Require (MATER), which is required for preimplantation embryo development beyond the 2-cell stage in mice. Human NLRP family contains 14 members. We identified 14 NLRP gene homologues and examined their spatial and temporal expression in rhesus macaque monkeys (Macaca mulatta). While all 14 NLRP genes are detectable in the macaque gonad, eight of them (NLRP2, 4, 5, 8, 9, 11, 13, and 14) are specifically or preferentially expressed in the ovary. In situ hybridization elucidated a specific oocyte expression pattern of the eight NLRP genes within the ovary. During the oocyte-to-embryo transition, seven of these oocyte-selective NLRP transcripts (excluding NLPR2) are enriched in maturing oocytes and early preimplantation embryos but diminish upon embryo genome activation, indicating an exclusive maternal origin of these transcripts. Though functionally unknown, the spatial and temporal distribution of these oocyte selective NLRP genes implies important roles of the NLRP family in oogenesis and early embryo development in nonhuman primates. PMID- 18509867 TI - In vivo model to determine fetal-cell enrichment efficiency of novel noninvasive prenatal diagnosis methods. AB - OBJECTIVE: To develop an in vivo model to determine fetal-cell enrichment efficiency of novel noninvasive prenatal diagnosis methods. METHODS: Efficiency of our three-step enrichment protocol was determined in vitro before fetal nucleated red blood cells (FNRBCs) were enriched from first-trimester maternal blood samples collected from the same patients pre- and postsurgical termination of pregnancy (TOP) (n = 10). FNRBCs enriched were identified using embryonic epsilon-globin immunocytochemistry and chromosomal fluorescence in situ hybridization. RESULTS: We recovered 37% of spiked FNRBCs (95% confidence interval (CI) 28.5-45.6; n = 8) in in vitro experiments. We show a consistent threefold increase in the number of epsilon + FNRBCs in maternal blood obtained immediately post-TOP (p = 0.005). A mathematical relationship was derived: observed number of pretermination primitive FNRBCs = 0.6 + 0.31 (coefficient between pretermination/post-termination primitive FNRBCs, 95% CI 0.12-0.49; p = 0.005) x observed number of post-termination primitive FNRBCs (R2 = 0.65). CONCLUSION: Our data demonstrate that maternal blood obtained immediately post TOP would be a good in vivo model to determine the enrichment efficiency of novel protocols and methods for noninvasive prenatal diagnosis. PMID- 18509868 TI - Is Mendelian randomization 'lost in translation?': comments on 'Mendelian randomization equals instrumental variable analysis with genetic instruments' by Wehby et al. PMID- 18509869 TI - The effect of gestational age on the outcome of second-trimester termination of pregnancies for foetal abnormalities. AB - OBJECTIVE: To explore the effect of gestational age on the outcome of second trimester termination of pregnancies for foetal abnormalities. METHOD: A retrospective study was conducted on 280 pregnancies terminated for foetal abnormalities in the second trimester using vaginal misoprostol. The gestational age at termination was divided into three groups: 13-16 weeks, 17-20 weeks and 21 23 weeks. The likelihood of (1) abortion within 24 h of commencement of misoprostol, (2) incomplete abortion and (3) experiencing significant side effects was compared among these three gestational groups after adjusting for maternal age, parity and body mass index (BMI). RESULTS: Compared to termination after 20 weeks, pregnancy termination for foetal abnormality before 17 weeks of gestation was associated with higher chance of incomplete abortion (OR 2.2, 95% CI 1.07-4.61, p = 0.032) and lower chance of experiencing significant side effects (OR 0.11, 95% CI 0.01-0.91, p = 0.041). CONCLUSION: Women undergoing pregnancy termination for foetal abnormalities in the early second trimester should be informed of possible higher chance of incomplete abortion. PMID- 18509870 TI - Structural determination of two new triterpenoids from Potentilla discolor Bunge by NMR techniques. AB - Two new pentacyclic triterpenoids, 3alpha, 30-dihydroxylup-20(29)-en-27-oic acid (1) and (20S)-3alpha, 29-dihydroxylupan-27-oic acid (2) were isolated from the whole herbs of Potentilla discolor Bunge. The structures of these two new compounds were elucidated, and complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, (1)H (1)HCOSY and ROESY). PMID- 18509871 TI - Regression of fetal cerebral abnormalities by primary cytomegalovirus infection following hyperimmunoglobulin therapy. AB - OBJECTIVE: To assess the effects of maternal and intra-amniotic hyperimmunoglobulin (HIG) infusions among cytomegalovirus (CMV) infected fetuses with ultrasound abnormalities following a primary CMV infection. PATIENTS AND METHODS: The subjects were fetuses with CMV-associated cerebral and other ultrasound abnormalities. Three mothers were treated with HIG infusions during pregnancy and two were untreated. Fetal ventricle size, organ echodensity and placental thickness were measured by ultrasound before and after HIG infusions. The children were evaluated between 3 and 7 years of age. RESULTS: The ventriculomegaly of all three fetuses of HIG-treated mothers regressed and the ascites, hepatic echodensities, periventricular echodensities, and intestinal echodensities disappeared. Their sensorial, mental and motor development was normal at 4, 4.7, and 7 years of age. In contrast, both infants born of untreated mothers had signs and symptoms of severe CMV cerebropathy. CONCLUSION: The outcomes of the infants born to HIG-treated mothers support the efficacy of HIG as a treatment for CMV-infected fetuses with ultrasound cerebral abnormalities. PMID- 18509872 TI - Attitudes toward the acceptability of reasons for pregnancy termination due to fetal abnormalities among prenatal care providers and consumers in Israel. AB - OBJECTIVES/BACKGROUND: Current prenatal diagnostic abilities confront parents and health professionals with complicated issues regarding termination of pregnancy (TOP) due to fetal abnormalities. 1. To assess and compare attitudes of consumers (women) and providers (health professionals) of prenatal care regarding TOP due to fetal abnormality.2. To identify factors related to these attitudes. METHODS: The study was conducted in southern Israel. Consumers (596) were interviewed by phone 5-8 weeks postpartum. Health professionals (351) filled out a self administrated questionnaire. RESULTS: More than half of the interviewees approved of TOP due to mental retardation, death during infancy, severe physical disability and very low quality of life (in descending order). For each condition, care providers were significantly more supportive of TOP than women, and had far fewer hesitations. The hierarchy of 'TOP acceptability' was similar in both populations. Factors associated with women's attitudes were degree of religiosity, Ashkenazi origin and country of birth. Two approaches toward TOP were identified: 'consistent' versus 'ad hoc'. CONCLUSIONS: Prenatal care providers and consumers differ in their attitudes regarding acceptability of reasons for TOP. Care providers offering prenatal tests should be aware of their patients' attitudes, in order to guide informed decisions regarding the tests. PMID- 18509873 TI - Family history of psychological problems in generalized anxiety disorder. AB - The current investigation examined self-reported family history of psychological problems in a large sample of individuals diagnosed with generalized anxiety disorder (GAD) and nonanxious controls. The GAD participants were all individuals receiving cognitive-behavioral therapy as part of two large randomized clinical trials. Family history information was obtained from the Anxiety Disorders Interview Schedule-Revised (ADIS-R; DiNardo & Barlow, 1988). The results indicate that, compared to control participants, individuals with GAD were more likely to have family members with anxiety problems, but not other psychological problems. Possible mechanisms for the familial transmission of GAD are discussed. PMID- 18509874 TI - Annual morphological cycles of testis and thumb pad of the male frog (Rana ridibunda). AB - In this study, Rana ridibunda was used as samples because of their wide use in Turkey. Male frogs were collected in the East Marmara region each month throughout 1 year. Frogs from every monthly collection were used to analyze structural components of the thumb pads and testes. Spermatogenetic activity of Rana ridibunda living in the East Marmara region was determined to be "potentially continuous" type. Generally, the increase in the number and the size of nuclei of Leydig cells was inversely proportional to the fluctuation of spermatogenetic activity. The lumen of the seminiferous tubules in testes contained, in addition to the spermatogenic cells, a Periodic-acid Schiff positive granular material. The amount of this material varied throughout the year, and that finding suggested a function related to spermiation. The components of thumb pads exhibited structural changes with respect to the activities of Leydig cells. During the periods where the Leydig cells were active, mucus glands (also called breeding glands) of thumb pads were also developed. On the other hand, we observed mixed glands with unknown function, which as first reported by us, and were poison glands in the thumb pads. The results suggest structural changes in the thumb pads are linked to changes in the testes. PMID- 18509875 TI - Mechanism of fibroblast-like synoviocyte apoptosis induced by recombinant human endostatin in rats with adjuvant arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by pronounced synovial hyperplasia, in which there may be an imbalance between the growth and death of fibroblast-like synoviocytes (FLS). The present study was undertaken to examine the effect of recombinant human endostatin (rhEndostatin) on FLS apoptosis in experimental RA. Adjuvant arthritis (AA) was induced in male Sprague Dawley (SD) rats. Using cultured AA FLS obtained from these rats, the apoptosis process was measured by terminal deoxyribonucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) as well as Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) labeling methods. In addition, the expression levels of the Fas, c-jun, NFkappaB, and caspase-3 gene products in synovial tissues were quantified by quantitative real-time polymerase chain reaction (qPCR) and/or Western blotting assays. Our data revealed that rhEndostatin induced apoptosis in AA FLS. The number and signal density of TUNEL positive cells were significantly increased in rats treated with rhEndostatin (2.5 mg/kg). The percentage of Annexin V-FITC-positive cells was 6.67% after treatment with rhEndostatin at 25 microg/mL for 48 hr, compared with only 3.32% among untreated control cells. There were significant increases in Fas mRNA, c jun mRNA, c-Jun protein, and caspase-3 (p20) protein in AA synovial tissues treated with rhEndostatin (2.5 mg/kg), whereas no significant difference in NFkappaB expression was detected between treated and untreated tissues. These findings indicate that rhEndostatin has a therapeutic effect on RA by inducing FLS apoptosis, which is strongly associated with increased expression of Fas, c jun, and caspase-3, but not NFkappaB. PMID- 18509876 TI - Antiviral effects of silymarin against hepatitis C: the jury is still out. PMID- 18509877 TI - Incidence of perforated diverticulitis and risk factors for death in a UK population. AB - BACKGROUND: Perforated diverticulitis (PD) remains a serious acute abdominal condition. The aims of this study were to measure its incidence in a large UK population and to identify factors affecting outcomes. METHODS: Computerized searches of hospital coding databases for PD were performed in five hospitals in East Anglia, UK. Data were collected from hospital records over 5 years (1995 2000). Incidence was calculated using population data, and factors associated with mortality and morbidity were identified using univariable and multivariable testing. RESULTS: Some 202 patients with PD were identified, of whom 93.1 per cent underwent surgery and 24.3 per cent died. The age-adjusted adult incidence of perforation was 3.5 per 100 000 per annum, with a standardized female to male ratio of 1.3 (95 per cent confidence interval (c.i.) 1.1 to 1.5) to 1. Risk factors for death were increased age (odds ratio (OR) 3.5 (95 per cent c.i. 1.9 to 6.1)), pre-existing renal disease (OR 18.7 (1.6 to 211.4)) and pre-existing use of non-steroidal anti-inflammatory drugs (NSAIDs) (OR 3.1 (1.3 to 7.3)). CONCLUSION: PD is uncommon, with the highest incidence in women over 65 years old. Mortality rates are high, particularly in those taking NSAIDs or with pre existing renal impairment. PMID- 18509878 TI - Data briefing. Prevention shows benefits across UK. PMID- 18509879 TI - Impact of clinical interview training on the empathy level of medical students and medical residents. AB - BACKGROUND: Empathy is a skill that can be acquired by practise and should become a habit. AIMS: Assess the impact of a communication skills workshop on the empathy level of medical students and medical residents. METHODS: Quasi experimental pre-test/post-test study. Empathy level was assessed in 203 subjects using the Jefferson Scale of Physician Empathy (JSPE), divided in two groups: one control group, and other experimental group that participated in an activity consisting of a 25-hour theoretical/practical workshop on communication and empathy. RESULTS: The mean pre-workshop JSPE score was similar in both groups. Post-workshop JSPE score increased 5.24 points (95 CI 3.82-7.09) (P<0.0001) in the experimental group, improving in 68.9% of participants. No significant increase in JSPE score after the second assessment was observed in the controls. For this difference the estimated effect size was 0.78. CONCLUSIONS: A communication skills workshop yields a slight improvement of crucial practice importance in subjects' empathy. PMID- 18509880 TI - Can medical students' pre-curriculum performance on the Ethics in Health Care Instrument (EHCI) be used as a predictor of future academic performance? A pilot study. PMID- 18509881 TI - The shadowing period--"could do better". PMID- 18509882 TI - Teaching hospitals vs teaching academies: the Bristol experience. PMID- 18509883 TI - Role of multi-drug resistance-associated protein-1 transporter in statin-induced myopathy. AB - This study investigated the effects of probenecid to inhibit the multi-drug resistance-associated protein-1 (MRP-1) in mediating the efflux and myotoxicity in rat skeletal muscles, with administration of rosuvastatin. Male Sprague-Dawley rats were administered daily, for 15 days, with either rosuvastatin (50, 100 or 200 mg/kg) or probenecid (100 mg/kg) alone, or with a combination of rosuvastatin (50, 100 or 200 mg/kg) and probenecid (100 mg/kg). Skeletal muscle toxicity was elevated with probenecid administered with 200 mg/kg/day of rosuvastatin, with the elevation of creatine kinase by 12-fold, alanine aminotrasferase by 10-fold and creatinine by 9-fold at day 15, with no adverse effects observed when probenecid was given alone. Mitochondria ultrastructural damage with enlargement, disruption, cristolysis and vaculation was seen in the soleus and plantaris of animals administered with probenecid and high dosages of statin. These muscles were also expressing more succinic dehydrogenase (SDH)-positive and cytochrome oxidase (CyOX)-positive fibers. Although generally well-tolerated, statins produce a variety of adverse skeletal muscle events. Hydrophilic statins, with reduced levels of non-specific passive diffusion rates into extra-hepatic tissues, are still seen to produce myopathy. This highlights the important roles of transport mechanisms in statin transport at the skeletal muscles. Excessive influx, reduced efflux or the combination of the two could result in elevated cellular levels of statins at the skeletal muscles, resulting in toxicity. This study provides preliminary evidence that the MRP-1 transporter and efflux at skeletal muscles possibly play significant roles in statin-induced myopathy. PMID- 18509884 TI - Risk factors for birth canal lacerations in primiparous women. AB - Lacerations of the birth canal are common side effects of vaginal birth. They are potentially preventable. Although serious long-term consequences have been identified for severe perineal lacerations, less attention has been paid to lacerations in other locations and how the risk factors vary for different lacerations. We analyzed a dataset including 1009 primiparous women with singleton pregnancies and vaginal deliveries, and we examined risk factors for third- and fourth-degree perineal lacerations and periurethral, vaginal, and labial lacerations using logistic regression analysis. Large fetal size (> or = 3500 g) substantially increased the risk of perineal (odd ratio [OR], 3.8; 95% confidence interval [CI], 1.8 to 7.9) and periurethral (OR, 2.3; 95% CI, 1.0 to 5.0) lacerations but not other types of lacerations. Episiotomy had no impact on perineal lacerations (OR 0.9) but had very strong protective effects for other lacerations (OR 0.1). Prolonged second stage of labor (> 120 minutes) increased the risk of perineal and vaginal lacerations but reduced the risk for periurethral lacerations. Instrumental deliveries were significant risk factors for third- and fourth-degree perineal lacerations, with by far the strongest effect for low forceps (OR 25.0 versus < 3 for outlet forceps, outlet vacuum, and low vacuum). We concluded that separating different birth canal lacerations is critical in identifying risk factors and potential preventive strategies. PMID- 18509885 TI - Multidisciplinary approach to congenital multiple arterio-porto-caval malformation: case report. AB - A case of intrahepatic arteriovenous malformation connected with umbilical, hepatic, portal, and arterial systems in a fetus diagnosed at 36 weeks of gestation is presented. Prenatal color Doppler ultrasonography in the fetal liver demonstrated complicated vascular connections fed by arterial and portal branches. Postnatal color Doppler, power Doppler, 3D power Doppler, abdominal multidetector computed tomography (MDCT), and angiography CT verified the prenatal diagnoses. Extended right hepatectomy was performed successfully on postnatal day 19. PMID- 18509886 TI - Oral ibuprofen for patent ductus arteriosus closure in preterm infants: does high osmolality matter? PMID- 18509887 TI - Burma: after the cyclone. PMID- 18509888 TI - Home DNA test kits cause controversy. PMID- 18509889 TI - Chemical biology meets systems biology. Introduction. PMID- 18509890 TI - Canadian Chemical Biology Network: biochemistry back to the future. PMID- 18509891 TI - The fear of old age. PMID- 18509892 TI - Multiple sulfatase deficiency in a Turkish family resulting from a novel mutation. AB - Multiple sulfatase deficiency (MSD) is an inherited lysosomal storage disease that affects post-translational activation of all of the sulfatases. Since biochemical and clinical findings are variable, the diagnosis is difficult in most of the cases. Missense, nonsense, microdeletion and splicing mutations in SUMF1 gene were found in all of the MSD patients analyzed. Here, we present clinical findings of two consanguineous patients with multiple sulfatase deficiency. They were found to be homozygous for a novel missense mutation c.739G > C causing a p.G247R amino acid substitution in the SUMF1 protein. PMID- 18509893 TI - Photosynthesis: CO2 uptake and the pathways of carbon fixation special issue. Preface. PMID- 18509894 TI - The health plan perspective of insomnia. PMID- 18509895 TI - [Angiosarcoma of the temporal bone]. AB - Angiosarcoma of the temporal bone is an extremely rare malignant tumor, which originates from vascular endothelium. We describe a case of a 57-year-old woman, initially presenting with progressive hearing loss, otorrhea as well as facial palsy diagnosed as a mastoiditis elsewhere. After subtotal mastoidectomy histological examination revealed an angiosarcoma of the mastoid. Since the tumor was not completely removed lateral petrosectomy and postoperative radiotherapy were performed. Afterwards the patient developed local recurrence with intracerebral tumor extension. During palliative polychemotherapy the patient developed a pneumonia and deceased. In this manuscript the morphology, imaging characteristics and current treatment options of angiosarcomas of the lateral skull base are reviewed and discussed. PMID- 18509896 TI - Connecticut's 2003 impaired-driving high-visibility enforcement campaign. PMID- 18509897 TI - The "healthy lifestyle guide pyramid" for children and adolescents. AB - INTRODUCTION: Increasing evidence demonstrates that risk factors for chronic diseases are established during childhood and adolescence. Consensus about the need to increase prevention efforts makes the adoption of a healthy lifestyle seem desirable from early childhood onwards. After reviewing educational tools for children and adolescents aimed at promoting a healthy lifestyle, it was recognized that there was a need to develop a simple educational tool specifically designed for these age groups. METHODS: Development of the healthy lifestyle pyramid for children and adolescents. RESULTS: We propose a three dimensional, truncated and staggered pyramid with 4 faces and a base, which introduces a completely new concept that goes beyond other published pyramids. Each of the faces is oriented towards achieving a different goal. Two faces (faces 1 and 2) are formulated around achieving a goal on a daily basis (daily food intake, face 1, and daily activities, face 2). Face 3 is an adaptation of the traditional food guide pyramid, adapted to children's energy, nutritional and hydration needs. Face 4 deals with both daily and life-long habits. On the base of the pyramid, there is advice about adequate nutrition alternating with advice about physical activity and sports. CONCLUSION: The Healthy Lifestyle Pyramid is specifically developed for children and adolescents according to current scientific knowledge and evidence-based data and includes easy-to-follow advice and full colour pictures. Following these guidelines should improve health and reduce risk factors, promoting enjoyable and appropriate development towards adulthood. PMID- 18509898 TI - [A pregnant woman with itching rash]. PMID- 18509899 TI - [Pregnancy, rash and itching]. PMID- 18509900 TI - The INSPIRE study: are different personality traits related to disease-specific quality of life (IBDQ) in distressed patients with ulcerative colitis and Crohn's disease? AB - BACKGROUND: To explore the relationship between personality and disease-specific quality of life [Inflammatory Bowel Disease Questionnaire (IBDQ)] in distressed [Perceived Stress Questionnaire (PSQ)] patients with ulcerative colitis (UC) and Crohn's disease (CD). METHODS: Included in the study were 56 patients with UC and 54 patients with CD ranging in age from 18 to 60 years with a relapse in the previous 18 months, a UC or CD activity index 4, a PSQ 60, and without serious mental or other serious medical condition. The patients completed the Buss-Perry Aggression Questionnaire, the Neuroticism and Lie (social conformity/desirability) scales of the Eysenck Personality Questionnaire, the Multidimensional Health Locus of Control (LOC) Scale [Internal (I), Powerful Other (PO), Chance (C)], the Toronto Alexithymia Scale, and the IBDQ. RESULTS: In linear regression controlling for sex, education (years), and clinical disease activity (AI) in separate analyses of UC and CD patients, higher IBDQ score was related to less social conformity in CD and less neuroticism in UC; higher emotional function score was related to less neuroticism in both CD and UC and less PO-LOC in UC. Higher social function score was related to less social conformity in CD and lower I-LOC and PO-LOC in UC. Bowel function and systemic symptoms were unrelated to personality in either UC or CD. CONCLUSIONS: Although the emotional function subscale was related to neuroticism in both UC and CD, the social function subscale and total IBDQ were related to different personality traits in UC and CD. Personality traits should be taken into account when using IBDQ in studies. PMID- 18509901 TI - [Top students will become dermatologists]. PMID- 18509902 TI - Ecological momentary assessment. AB - Assessment in clinical psychology typically relies on global retrospective self reports collected at research or clinic visits, which are limited by recall bias and are not well suited to address how behavior changes over time and across contexts. Ecological momentary assessment (EMA) involves repeated sampling of subjects' current behaviors and experiences in real time, in subjects' natural environments. EMA aims to minimize recall bias, maximize ecological validity, and allow study of microprocesses that influence behavior in real-world contexts. EMA studies assess particular events in subjects' lives or assess subjects at periodic intervals, often by random time sampling, using technologies ranging from written diaries and telephones to electronic diaries and physiological sensors. We discuss the rationale for EMA, EMA designs, methodological and practical issues, and comparisons of EMA and recall data. EMA holds unique promise to advance the science and practice of clinical psychology by shedding light on the dynamics of behavior in real-world settings. PMID- 18509903 TI - Move over proteomics, here comes glycomics. PMID- 18509904 TI - Mucosal matters. Foreword. PMID- 18509905 TI - Re: Prediction of liver fibrosis in patients with chronic hepatitis B. PMID- 18509906 TI - [Quantitative measures for assessing the functional state of the human body during diagnostic procedure]. AB - Two approaches to calculation of the qualitative measures for assessing the functional state level of human body are considered. These approaches are based on image and fuzzy set recognition theories and are used to construct diagnostic decision rules. The first approach uses the data on deviation of detected parameters from those for healthy persons; the second approach analyzes the degree of deviation of detected parameters from the approximants characterizing the correlation differences between the parameters. A method for synthesis of decision rules and the results of blood count-based research for a number of diseases (hemophilia, thrombocytopathy, hypertension, arrhythmia, hepatic cirrhosis, trichophytia) are considered. An effect of a change in the functional link between the cholesterol content in blood and the relative rate of variation of AST and ALT enzymes in blood from direct proportional (healthy state) to inverse proportional (hepatic cirrhosis) is discussed. It is shown that analysis of correlation changes in detected parameters of the human body state during diagnostic process is more effective for application in decision support systems than the state space analysis. PMID- 18509907 TI - [Molecular biology methods used in the study of Mycobacterium tuberculosis]. PMID- 18509908 TI - [Experience with bronchomunal used in the combined treatment of patients with bronchial asthma and chronic obstructive pulmonary disease]. AB - The paper presents the results of treatment in 30 patients aged 16-59 years who have bronchial asthma and chronic obstructive pulmonary disease, by using a bacterial vaccine (bronchomunal) containing antigens of opportunistic bacteria: Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus viridans, Streptococcus piogenes, Moraxella catarrhalis. Immunotherapy with the vaccine had good and excellent effects in 73.33 of cases; the mean duration of acute respiratory viral infection decreased from 16 to 9 days after vaccination and a need for antibiotics. In the comparison group, a good effect was noted in 40% of the patients during one-year follow-up; the difference was statistically significant. The vaccine's tolerance was good; only 3 (9.9%) patients were observed to have vaccination-induced complications: exacerbations of chronic maxillary sinusitis and chronic bronchitis in 2 and 1 patients, respectively. The positive effect of bronchomunal was associated with the better values of cellular immunity, stabilized phagocytosis, and lower IgE levels. PMID- 18509909 TI - [Epidemiology of tuberculosis and current approaches to improving measures against tuberculosis among adolescents]. AB - The specific features of the epidemiological process and clinical forms of tuberculosis in Vladivostok adolescents treated in 1999 to 2006 were studied. Two periods were identified: 1) from 1999 to 2002 when fluorographic studies were annually made and 2) from 2003 to 2006 when fluorography was carried out every two years. It was established that during Period 2, there was a rise in tuberculosis morbidity among adolescents, which was attended by an increase in the number of patients with open and destructive forms. Adolescents aged 17 years, girls, first-year students were found to be more frequently ill. 41.1% of the patients underwent a fluorographic study less than a year ago. The foregoing generates the necessity of submitting the medical documentation of enrollees and first-year students along with the data of a fluorographic study and tuberculin tests. PMID- 18509910 TI - [The specific features of immunity and immunological responsiveness in elderly and senile patients with tuberculous pleurisy]. AB - The paper gives the results of immunological and clinical studies made in 162 elderly and senile patients with tuberculous exudative pleurisy and a control group of 162 tuberculous pleurisy aged 20-40 years. The elderly and senile patients were found to have more significant immunological responsiveness impairments, suppressed T-cell immunity, and inhibited pleural granulomatous reactions than in the young patients. These alterations substantially worsen the course and outcome of the disease and require im munological correction. PMID- 18509911 TI - [The values of the lipid peroxidation-antioxidative defense system as predictors of the poor course of infiltrative pulmonary tuberculosis]. AB - The authors have studied whether an indirect clinical-and-laboratory assessment of the extent of a destructive process in the tuberculous infiltrates and that of concomitant pleural affection can be made from the values of the lipid peroxidation-antioxidative defense (PLO-AOD) system in the blood of patients with infiltrative pulmonary tuberculosis (IPT). Regression analysis has indicated that PLO-AOD values significantly reflect the extent of X-ray verified destructions in the tuberculous infiltrates, concomitant pleural involvements, the specific features of immunity of patients with IPT. The neural network technology used to analyze the baseline PLO-AOD values has been ascertained to permit a reliable prognosis of decay cavity closure 4 months after the initiation of medical treatment and of a need for surgical treatment for IPT. PMID- 18509912 TI - [Polymerase chain reaction to determine and control drug-resistant Mycobacterium tuberculosis strains]. AB - Real-time polymerase chain reaction was used to develop a one-stage procedure for molecular genetic analysis of Mycobacterium tuberculosis (MBT) DNA in order to determine mutations associated with drug resistance to the antituberculous agents: isoniazid and rifampicin. To analyze the spread of drug-resistance of the causative agent of tuberculosis in Russia, two thousand MBT strains were studied in 24 regions of all the federal districts. Testing 1406 MBT strains isolated by first detected and untreated patients revealed multidrug resistance (MDR) in 21.9% of cases. MRD was detected in 58.5% of the previously treated patients with MDR. The agreement of molecular genetic analysis of drug resistance with the results of cultural tests of 1096 strains was 94%. PMID- 18509913 TI - [Experience in comprehensively assessing the health status of the Saint Petersburg population]. AB - A scheme is proposed to calculate the overall human health indices by the values of partial grouped parameters: demographic, environmental, environment-induced, total (including social) morbidity; socioeconomic and preset threshold levels of their qualitative assessment. This method has been used to estimate morbidity rates in 7 Saint Petersburg districts. PMID- 18509914 TI - [Impact of man-made atmospheric pollution on the incidence of congenital malformations]. AB - S u m mary. The paper presents the results of studying the qualitative and quantitative representation of anthropogenic pollutants in the ambient air environment of Belgorod and evaluating their impact on the prevalence of neonatal congenital malformations. The level of atmospheric pollutant emissions is shown to vary by an average of 4 to 10 times in some areas. The vast majority (91.7%) of pollutants form clusters (n = 11), suggesting their combined influence. Each group of clusters includes 2 to 9 pollutants (median 4.64 pollutants). Atmospheric pollutants have been ascertained to adversely affect the prevalence of congenital malformations in children. A multiple regression equation has been derived to predict the frequency of congenital malformations, by taking into account the qualitative and quantitative spectrum of atmospheric pollutants. PMID- 18509915 TI - [Hygienic aspects of iodine deficiency diseases in the Tomsk Region]. AB - The impact of the content of iodine and other trace elements in the environmental objects of the Tomsk Region on the development of iodine-deficiency diseases was studied in children. Slightly sour soils and low soil humus levels in the Tomsk Region were ascertained to be conducive to low iodine content in soils and plants. As compared with the clarke of iodine in the lithosphere, the soils in the Tomsk Region were marked by low iodine levels, permitting it to be, in terms of endemic goiter, referred to as a biogeochemical province. The soils of rural areas in the Tomsk Region were characterized by the specific trace element composition--the high content of molybdenum and copper and the low level of zinc, which is a factor that is of importance in the development of endemic goiter. In children, daily dietary requirements for iodine were met at 60% of the normal levels, those for animal proteins at 50%, at the same time there was deficiency of amino acid tyrosine, the substrate involved in the synthesis of thyroid hormones. Endemic goiter occupies 80% in the structure of thyroid diseases. There is a considerable predominance of early goiter forms over severer ones. In terms of the level of ioduria, 64% of the examined children had mild iodine deficiency. PMID- 18509916 TI - [Influence of poor factors of airports on human health]. AB - The hygienic study conducted in Rostov-on-Don has shown that air transport and airports are important sources of physical and chemical pollution of the environment. Human health examinations served to illustrate the adverse impact of airports on the environmental and hygienic living conditions of the population. The performed studies provided the basis for purpose-oriented program to enhance the environment and to reduce morbidity rates in accordance with the National Environmental Hygiene Program. The developed algorithm of the assessment and reduction of a risk for diseases under the influence of poor factors associated with the activities of airports is designed to provide the authorities and concerned organizations with information, to make managerial decisions, and to work out health-improving measures. PMID- 18509917 TI - [Evaluation of the toxicity and hazard of waste resulting from municipal sewage purification and sediment incineration]. AB - Municipal waste water, sand box sewage sludge, floating substances, raw tank sediments, biological solids, a mixture of compacted and dewatered sediments, flue gases, ashes, and smoke waste products (GOST 12.1.007-76) are referred to as hazard classes 3 and 4. The authors consider the conditions contributing to the occurrence of acute and chronic intoxications of the personnel, lung fibrous lesion, infectious diseases, helminthic invasions, environmental pollution at the basic stages of municipal waste treatment and sediment utilization, PMID- 18509918 TI - [The health status of stewardesses and its influencing factors]. PMID- 18509919 TI - [Environmental factors and parkinsonian syndrome]. AB - Particular emphasis has been in recent years placed on the study of an integrating role of medicine in the prevention, early diagnosis, and treatment of diseases associated with the influence of environmental factors. Among nervous system diseases associated with the adverse influence of environmental factors, the parkinsonian syndrome has been the particular concern, which is due to its higher incidence rates and early disability in able-bodied persons. In this connection, the purpose of the study was to review the data available in the literature on the neurotoxicity of heavy metals and other environmental factors causing toxic parkinsonism. The mechanisms of development and the clinical manifestations of the parkinsonian syndrome were considered in occupational manganese poisonings, magnesium deficiency, and carbon monoxide intoxication. Profound assessment of the influence of environmental factors on certain population groups is possible only when hygienists, toxicologists, biochemists, immunologists, molecular geneticists, and clinicians combine their efforts. PMID- 18509920 TI - [Prevention of late consequences of mutagenic exposure on workers]. AB - The staff of the branches of industry, which are followed up by the Federal Biomedical Agency (FBMA) Agency of Russia, is constantly exposed to chemical and radioactive agents in small doses both during its work at these enterprises and due to atmospheric emissions. These agents can cause mutations in human somatic and germ cells, which may in turn increase the risk of cancer and congenital abnormalities. Only complex prevention of late consequences of mutagenic exposure on the health status will make it possible to reduce the incidence of malignancies and congenital and hereditary diseases in the staff and the general population. The list of their primary prevention measures proposed to be introduced into the system of the Federal Biomedical Agency of Russia. PMID- 18509921 TI - [The eye analyzer in machine-building industry workers according to electrophysiological indices]. AB - To study visual analyzer electrophysiological changes in workers exposed to occupational physical factors remains to be a topical problem. The authors investigated retinal electrosensitivity and optic nerve electrolability and performed electroretinography in this group of workers, which determined significant retinal functional changes typical of retinal metabolic disturbances in workers having a long service. PMID- 18509922 TI - [Evaluation of sanitary-and-epidemiological safety of medical wares]. AB - The medical personnel operating currently available medical equipment can be exposed to physical factors. To prevent the adverse consequences of these exposures, medical equipment undergoes sanitary-and-epidemiological examination, resulting in the drawing up of a sanitary-and-epidemiological report. For regulation of this procedure, a management directive has been worked out, which determines an examination procedure, a hygienic classification of medical equipment, allowable values for controlled safety indices during operation, etc. Introduction of the directive makes it possible to upgrade the quality of sanitary-and-epidemiological equipment audit, to improve working conditions, and to keep health in the personnel operating the present-day high-technology equipment. PMID- 18509923 TI - [Provision of Russia's pediatric population with sanitary-and-epidemiological well-being]. AB - There have been negative tendencies for the formation of public health to be preserved in the past decade. The most important tendency is for the worse physical development and health of children and adolescents. The 2002 All-Russian medical examination of children has provided support for the negative tendencies in the health status of children - the proportion of healthy children during the above period had reduced from 45.5% to 33.9%. The incidence of endocrine, digestive, and locomotor diseases, congenital malformations, and mental disorders has particularly increased. It should be said that the health status of the general population and primarily children is determined by the influence of a number of factors, including sociohygienic living conditions (up to 25-40%), man made environmental pollution (above 25%), intraschool environmental factors (21 27%), and health care quality (up to 25%). School-age children account for about 70% of the pediatric population. It is in this age group that shows the greatest rise in morbidity. In recent years, the Principal Public Sanitary Inspector of the Russian Federation has published 5 regulations on various aspects of sanitary and-epidemiological well-being in children. A number of regions of the country take actions on the creation of proper conditions for the upbringing, education, and health-improvement of children. The present-day educational system is characterized by a great diversity of curriculums. At the same time, only one curriculum and one educational technology have a sanitary-and-epidemiological conclusion on health safety. Adolescents with risky forms of sexual behavior have increased in number. This in combination with underhygienic measures leads to the spread of HIV infection and other sexually transmitted diseases among them. For children's sanitary-and-epidemiological well-being in the Russian Federation, it is expedient: 1) to complete the development of the adverse influence of environmental factors on the health status of children, which envisages the implementation of measures to improve their habitat (ambient air, portable water), intraschool environment, and living conditions in the families, socially poor ones in particularly, nutritional pattern, and lifestyle in children and adolescents; and 2) to introduce prophylactic syllabi at school in order to form healthy life style skills, a conscious attitude to the health status of theirs and those around them, etc. in children and adolescents. PMID- 18509924 TI - [Efficiency of an arbitrary respiratory rhythm reducing method in students of special medical physical educational groups]. AB - Three-month decreased voluntary optimum respiration rate (DVORR) trainings during therapeutic exercises in students with various abnormalities caused increases in vital capacity and chest circumference amplitude. DVORR also affected cardiovascular performance. During exercises, heart rate decreased and pulse pressure increased, suggesting economized cardiac performance and better myocardial function. The use of this method in the students reduces morbidity, improves health, and increases physical fitness and progress in studies. PMID- 18509925 TI - [Sanogenetic effects of thalassotherapy in medical personnel of a surgical emergency team]. AB - Significant differences in nervous and cardiovascular systems state and humoral homeostasis indices were established in relative healthy personnel (62 persons) of Resuscitation Unit and Burn Center in comparison with control group (60 persons). Thalassotherapy has obvious health-improving effect in asthenia clinical symptoms regress, decrease in number of acute respiratory diseases and otorhinolaryngological organs chronic pathology relapses. Sanogenetic effects are realized via normalization of level of cortisol, thyroid-stimulating hormone, immune sate indices and optimization of lipid peroxydation processes. PMID- 18509926 TI - [Possibilities of respiratory exercises in medical rehabilitation of preschool age children with respiratory organs pathology]. AB - The purpose of the work is to study efficiency of a new medical technology in rehabilitation of children with respiratory organs pathology. 177 children at the age from 2 to 7 years were randomized into two groups: 90 children of the main group performed the complexes of physical and respiratory exercises according to developed medical technology, 87 children of the control group performed standard physical exercises on physical training classes. The results of the study revealed decrease in frequency of acute respiratory disease in 1.83 times, exacerbation of bronchial asthma in 1.86 times in children of the main group. Improvement in development of physical qualities and spirography indices against the background of physical and respiratory exercises according to developed medical technology was detected. Significant cytoprotective action to nasal mucous tunic in preschool age children with respiratory organs pathology was registered. PMID- 18509927 TI - [Quantum-wave physiotherapy in the treatment of myopia in children]. AB - It is established that cyclicity of excitation and inhibition processes in healthy persons is symmetric, 11.9+0.2 cycles in 1 minute, and reflects structural (retina nerve fibers layer thickness) and functional (visual acuity) symmetry of pair eyes, autonomous regulatory system dynamic balance as the etalon of health. Myopia is accompanied by autonomous visual cycles acceleration up to 14.3+2.4 cycles in 1 minute at low degree and up to 17.7+2.2 cycles in 1 minute at mid degree with their rhythmicity disturbance. Simultaneously asymmetrical thinning of retina nerve fibers layer was noted. Bilateral electroconductivity on meridians on the right side was lower than on the left side, which characterized vegetative balance asymmetry as pathology index. Quantum-wave physiotherapy imitates autonomous cyclicity of visual system and recovers vegetative balance bilateral symmetry and structural-functional symmetry of visual organ with the increase of visual acuity in 1.5 times. PMID- 18509928 TI - [Management of a spa complex based on systems laws and principles]. PMID- 18509929 TI - [Ultraviolet irradiation of skin combined with balneotherapy in the treatment of patients with atopic dermatitis]. PMID- 18509930 TI - [Natural curative resources of Samara region]. PMID- 18509932 TI - The perils of dental tourism, closer to home. PMID- 18509933 TI - Measuring body fat and linking measurements to metabolic syndrome. PMID- 18509934 TI - Recognizing signs of heart attack and taking action. PMID- 18509935 TI - Low levels of magnesium place diabetics at increased risk. PMID- 18509936 TI - How blood pressure affects the aging mind. PMID- 18509937 TI - Prevalence of antibodies to Toxoplasma gondii infection in humans and domestic animals in Samsun province, Turkey. PMID- 18509938 TI - Prevalence and mortality rate of peste des petitis ruminant (PPR): possible association with abortion in goat. AB - Present study was designed to investigate the prevalence and mortality (%) caused by Peste des Petitis Ruminant (PPR) and its possible association with abortion in goat flocks at different areas of Pakistan. A total of 140 animals were samples in the population of 650 which was having 185 deaths (Mortality rate = 28 %) from three different regions of the country. There were 58 abortions in the 140 pregnant goats of above said population One hundred & ten (110) serum samples from diseased, recovered and apparently healthy animals were tested for the presence of PPR antibodies by competitive ELISA (c ELISA). Eighty-four (84) animals were positive for PPR antibodies whereas in apparently healthy adult goats in the same flock, no PPR antibodies were detected. Twenty-four (24) tissue samples collected from the dead animals and six samples from aborted fetus were tested for the presence of PPR antigen by Immuno-capture ELISA (Ic ELISA). Nineteen (19) out of thirty (30) organ samples mainly from lung, spleen, lymph node were found positive for PPR antigen but negative from lungs of aborted fetus. There was a high rate of abortions (28-45%) in each of the outbreak and it was highest in the outbreak of Golra Sharif, Islamabad (No. = 21 in total population of 100). As the serum samples from the aborted dams were found positive for PPR antibodies so the study provides the possible association of mortality and prevalence of PPR disease with high rate of abortions in goat. PMID- 18509939 TI - Detection of Leptospira and Brucella genomes in bovine semen using polymerase chain reaction. PMID- 18509940 TI - The prevalence of gastrointestinal helminth infections in pigs in Kenya. AB - The prevalence of helminth infection, species spectrum and worm burdens in Kenyan pigs was examined. A total of 115 gastrointestinal tracts (GIT) from 61 growers and 54 adult pigs were examined between February 2005 and January 2006. Seventy eight (67.8%) had one or more helminth parasites, of which thirty six (31.3%) were mixed infection. Ten types of helminth parasites encountered in descending order of prevalence were, Oesophagostomum dentatum (39.1%), Trichuris suis (32.2%), Ascaris suum (28.7%), Oesophagostomum quadrispinulatum (14.8%), Trichostrongylus colubriformis (10.4%), Trichostrongylus axei (4.3%), Strongyloides ransomi (4.3%), Hyostrongylus rubidus (1.7%), Ascarops strongylina (1.7%) and Physocephalus sexalutus (0.9%). Oesophagostomum dentatum was the most prevalent species (51.9%) in the adult pigs, while Trichuris suis was the most prevalent species (44.3%) in growers. The highest worm counts were recorded in the out door production system. Oesophagostomum quadrispinulatum, Trichostrongylus colubriformis, Trichostrongylus axei, Hyostrongylus rubidus, Ascarops strongylina and Physocephalus sexalutus were recorded in Kenya for the first time. The high prevalence and wide spectrum observed in the present study suggests that helminth infection may be a constraint to economic pig production in the country and there is need to institute control measures. PMID- 18509941 TI - Proliferation and transmission patterns of Pasteurella multocida B:2 in goats. AB - This report describes the proliferation and transmission patterns of Pasteurella multocida B:2 among stressful goats, created through dexamethasone injections. Thirty seven clinically healthy adult goats were divided into three groups consisted of 15 goats in group A, 11 goats in group B and the remaining 11 in group C. At the start of the study, all goats of group A were exposed intranasally to 1.97 x 10(10) CFU/ml of live P multocida B:2. Dexamethasone was immediately administered intramuscularly for 3 consecutive days at a dosage rate of 1 mg/kg. The exposed goats were observed for signs of HS for a period of 1 month. At the end of the 1-month period, 11 goats from group B were introduced into and commingled with the surviving goats of group A before all goats from both groups were immediately injected intramuscularly with dexamethasone for 3 consecutive days. The treatment with dexamethasone was then carried out at monthly interval throughout the 3-month study period. Goats of group C were kept separately as negative control. Three surviving goats from each group were killed at 2-week interval for a complete post-mortem examination. Two (13%) goats of group A were killed within 24 hours after intranasal exposure to P multocida B:2 while another two (13%) goats from the same group were killed on day 40, approximately 10 days after the second dexamethasone injection. All four goats showed signs and lesions typical of haemorrhagic septicaemia. Bacteraemia was detected in 3 goats of group A that were having rectal temperature higher than 41degrees C. The P. multocida B:2 isolation pattern was closely associated with dexamethasone injections when significantly (p < 0.05) higher rate of isolations from both groups were observed after each dexamethasone injection. Transmission of P multocida B:2 from goats of group A to group B was successful when P multocida B:2 was isolated from goats of group B for a period of 28 days. There was a strong correlation between dexamethasone injections, rate of bacterial isolation and serum cortisol level. The IgG level showed an increasing trend 2 weeks after exposure to P multocida B:2 and remained high throughout the study period. PMID- 18509942 TI - Comparison of two milk pricing systems and their effect on milk price and milk revenue of dairy farms in the Central region of Thailand. AB - A study was conducted to investigate determinates of how milk pricing system, farm location, farm size, and month and year affected farm milk price (FMP), farm milk revenue (FMR) and loss in FMR of dairy farms in the Central region of Thailand. A total of 58,575 milk price and 813,636 milk yield records from 1034 farms were collected from November of 2004 to June of 2006. Farms were located in the districts of Muaklek, Pak Chong, Wang Muang, and Kaeng Khoi. A fixed linear model was used to analyze milk price of farms. Two pricing systems were defined as 1 = base price plus additions/deductions for milk fat percentage, solids-non fat, and bacterial score, and 2 = same as 1 plus bulk tank somatic cell count (BTSCC). Farm size (small, medium, and large) was based on the number of cows milked per day of farms. Results showed that FMP were lower (P < 0.05) in pricing system 1 than pricing system 2. Most small farms had higher (P < 0.05) milk prices than medium and large farms across both pricing systems. Large farms lost more milk revenue due to deductions from bacterial score and BTSCC than small and medium farms. PMID- 18509943 TI - Ostrich (Struthio camelus) production in Egypt. AB - This review discusses the historical, developmental and practices of ostrich farming in Egypt. In the early 20th century, ostrich farming was very important for production of ostrich feathers and documents were produced to perfect the art of procuring the plumes from the birds and subsequently processing them. Pharaohs used ostrich feathers for adornment. Of 43 provinces, 12 were featured in 2003 2004 as farming ostriches: Alexandria, Al-Behera, Al-Dakahlia, Al-Wadi Al-Gadid, Aswan, Cairo, El-Sharkia, Geiza, Ismailia, Kafr-El-Sheikh, Matrouh and Nubaria. Abattoirs and tanneries specialising in ostrich handling are limited to two. Egypt has numerous strengths and opportunities to develop its ostrich sector. Rising meat prices suggest that fresh ostrich meat is unaffordable to many locals. Funds may be allocated to local advertising campaigns to promote ostrich meat; provision of incentives to farmers; and improving the capacity of abattoirs. PMID- 18509944 TI - Endemic status of Trypanosoma evansi infection in a horse stable of eastern region of India--a field investigation. AB - Diagnosis of Trypanosoma evansi infection in a horse stable of Eastern Region of India on the basis of examination of Giemsa stained blood smears have been done. A high percentage (12.74%) of horses of this stable was found suffering from T evansi infection. This high prevalence of T evansi in horses, in this area could be considered as an alarming situation which has never been explored previously in horses of Eastern Region of India. After a period of 2 months and 18 days of treatment with quinapyramine sulphate and quinapyramine chloride, reinfection with T evansi in treated horses of this stable were noticed. Clinical signs of affected horses and possible causes of reinfection have been discussed. PMID- 18509945 TI - Rapid detection of Peste des Petits Ruminants (PPR) virus antigen in Sudan by agar gel precipitation (AGPT) and haemagglutination (HA) tests. AB - AGPT and HA tests were employed for rapid diagnosis of PPRV infection in sheep and goats in Sudan. Forty lymph nodes and spleen samples from suspected cases of PPR in both sheep and goats were examined by AGPT and HA tests for detection of PPRV antigen. Viral antigen was detected from (77.5%) of the samples tested by AGPT and (92.5%) tested by HA test. The results of both tests revealed that HA test was more sensitive than AGPT for detection of PPRV antigen (Kappa statistics 0.4366). Another advantage of the HA test over AGPT was that it can differentiate PPRV from RPV. Thus the HA test represents a quick, easy, simple, cheap and reliable confirmatory test for the diagnosis of PPR and differential diagnosis of PPRV and RPV. The HA test was carried out using chicken, goat and pig RBCs. Chicken RBCs were found to be the most sensitive for detection of PPRV antigen, followed by goat then pig RBCs. The HA time when using chicken RBCs was 20-25 minutes, using goat RBCs was 25-30 minutes and using pig RBCs was 40-45 minutes. The distribution of PPR infection in four different regions of Sudan was investigated. PMID- 18509946 TI - Calf morbidity and mortality in smallholder dairy farms in Ada'a Liben district of Oromia, Ethiopia. AB - A longitudinal observational study on calf morbidity and mortality was conducted in smallholder dairy farms in Ada'a Liben district of Oromia, Ethiopia. A total of 185 calves from 112 market oriented smallholder dairy farms were selected randomly and regularly monitored for clinical health problems up to six months of age. Information on potential risk factors was collected by personal observation during the regular visit to farms and from questionnaire survey conducted during the study period. The overall incidences of crude morbidity and crude mortality were 62% and 22%, respectively. The most frequent disease syndrome was calf diarrhea with the incidence of 39% followed by joint ill 6%. The other disease conditions/syndromes diagnosed include navel ill, pneumonia, septicemic conditions, congenital problems and miscellaneous cases. Age of the calves, age at first colostrum ingestion and cleanness of the calf barns significantly influenced morbidity. Older calves (greater than three months of age) were at lower risk of crude morbidity than younger calves (less than three months of age) (HR = 0.42, P = 0.001). Higher risk of crude morbidity was observed in calves that ingested their first colostrum meal later than 6 hours of age compared to those that ingested colostrum earlier (HR = 2.24, P = 0.001). Similarly, calves housed in unclean barns were at higher risk of morbidity than calves housed in clean barns (HR = 1.75, P = 0.024). Of the 20 potential risk factors investigated, age was the only factor that was found significantly associated with mortality (HR = 0.04, P = 0.001). Calves older than three months of age were at lower risk of mortality than younger calves. PMID- 18509947 TI - Commercial crocodile farming in Botswana. AB - A survey-based study was carried out to assess the state of crocodile farming in Botswana. A prepared, structured questionnaire was dispatched to crocodile farmers based on a directory provided by the Fisheries section of the Department of Wildlife and National Parks in the Ministry of Environment, Wildlife and National Parks. The oldest farm was established in 1986. An average of three farms have been in operation since then, all of which obtained their stock from the Okavango and Thamalakane rivers in Botswana. The current stock averages 5,419 animals as follows: breeders 4%, hatchlings 56%, and growers 40%. The average clutch size and average hatchability were 47 eggs/clutch and 67% respectively. Mortality among hatchings and growers averaged 8.3% up to 12 weeks of age. Only one farm encountered some problems with Salmonella and fungal infections of the belly. Raw skins are sold to South Africa as a result of the absence of a tannery. Crocodile farming should be encouraged in Botswana since a good market for crocodile products already exists. PMID- 18509948 TI - [Intestinal parasitoses and worm infestations]. PMID- 18509949 TI - Prenatal diagnosis. Preface. PMID- 18509950 TI - Response to Sheldon Watts: "Yellow fever immunities in West Africa and the Americas in the age of slavery and beyond.". PMID- 18509951 TI - Leading the way to change. PMID- 18509952 TI - Market solutions for social problems: working-class housing in nineteenth-century London. PMID- 18509953 TI - Vermont's high court considers pets'special value. PMID- 18509954 TI - Study links occupational exposures with risk of miscarriage. Anesthetic gases, radiation, pesticides are areas of concern for pregnant workers. PMID- 18509955 TI - Petites bourgeoises and penny capitalists: women in retail in the Lille area during the nineteenth century. PMID- 18509956 TI - Deborah Charlesworth. PMID- 18509958 TI - More on organic farming systems. PMID- 18509957 TI - Scientific organizations as agents of change: the Palestine Exploration Fund, the Deutsche Verein zur Erforschung Palastinas and nineteenth-century Palestine. PMID- 18509959 TI - Thoughts on solutions to pet obesity. PMID- 18509960 TI - Scientists and the cultural politics of academic disciplines in late 19th-century Germany: Emil DuBois-Reymond and the controversy over the role of the cultural sciences. PMID- 18509961 TI - [Techniques and current status of autologous liver transplantation]. PMID- 18509962 TI - [Clinical study of bone tunnel expansion in anterior cruciate ligament reconstruction]. AB - OBJECTIVES: To measure the size and change of bone tunnel in arthroscopic assisted anterior cruciate ligament (ACL) reconstruction with hamstring tendon autograft by X-ray, and evaluate the incidence, extent, shape and reasons of the bone tunnel expansion and analyze the relationship between bone tunnel expansion and clinical results. METHODS: Fifty-one cases of arthroscopic ACL reconstruction with hamstring tendon autograft were performed, and they were followed up at average of 16 months postoperatively. The diameter, shape, position and angle of femoral and tibial bone tunnel were measured using X-ray, and other clinical information had been collected including gender, age, method of tibial fixation, range of motion, KT 2000 and evaluation of muscle strength and so on. All data were entered into the computerized relational database to analyze and compare using the chi square test and correlation analysis. RESULTS: The incidence of tunnel expansion after ACL reconstruction was 85%-94% in femoral tunnel and 65% in tibial tunnel. The extent of tunnel expansion was 51%-53% in femoral tunnel and 40%-44% in tibial tunnel. The most common shape of tibial tunnel was type O in the A-P X-ray view and type V in the lateral X-ray view. Femoral tunnels anterior to the expected ones were more likely to enlarge. Tibial tunnels anterior to the expected ones were easier to expand. An acute tibial or femoral tunnel angle could result in the femoral tunnel expansion. CONCLUSIONS: The incidence and extent of bone tunnel expansion in arthroscopic ACL reconstruction with hamstring tendon autograft are more significant in femoral tunnel than in tibial tunnel. Bone tunnel expansion is correlated to patients' age, height, BMI and location of the tunnel. The main factors related to bone tunnel expansion are the location and angle of the tunnel. PMID- 18509963 TI - [Arthroscopic simultaneous reconstruction of posterior cruciate ligament using double femoral tunnel technique and anterior cruciate ligament with achilles allograft]. AB - OBJECTIVES: To introduce the technique of arthroscopic simultaneous reconstruction of posterior cruciate ligament (PCL) using double femoral tunnel, single-bundle transtibial tunnel PCL technique and anterior cruciate ligament (ACL) with achilles allograft, and to evaluate the clinical outcome. METHODS: Fourteen patients with PCL and ACL injuries after a minimum follow-up 18 months were received. Arthroscopically assisted simultaneous ACL/PCL reconstruction with achilles allograft were performed using the single-incision endoscopic ACL technique and the double femoral tunnel, single-bundle transtibial tunnel PCL technique. The Lysholm and Tegner knee score scale were used for functional evaluation. All patients were evaluated with physical examination and KT-1000 arthrometer testing. The mean knee flexion was (123.6 +/- 2.5) degrees preoperatively. The Lysholm score was 52.8 +/- 2.2. The Tegner score was 5.9 +/- 0.5 before injury, 1.2 +/- 0.9 preoperatively. RESULTS: The mean time from injury to the reconstructive procedure was 19.5 d. The mean knee flexion was (117.9 +/- 2.8) degrees postoperatively( t = 1.54, P = 0.14). As to the Lachman test for 14 patients, the results of 13 patients (92.9%) was negative. As to posterior drawer test, the results of 12 patients (85.7%) was negative. The Lysholm score was 92.9 +/- 3.3 at final evaluation (t = 17.009, P < 0.001). KT-1000 arthrometer testing at 25 degrees knee flexion showed that the side-to-side difference was below 2 mm in 9 cases, 3-5 mm in 4 cases, 6 mm in 1 case. At 75 degrees knee flexion the difference was below 2 mm in 10 cases, 3-5 mm in 3 cases, 6 mm in 1 case. The Tegner score was 5.4 +/- 0.8 at final evaluation. The difference between the preoperative score and the postoperative was statistically significant (F = 4.2, P < 0.01). CONCLUSIONS: Combined ACL and PCL injuries can be successfully treated with arthroscopic simultaneous reconstruction of PCL using double femoral tunnel technique and ACL with achilles allograft. The double femoral tunnel technique more closely approximates the anatomic insertion the native PCL. Most patients recover a functionally stable knee. PMID- 18509964 TI - [The clinical results of anterior cruciate ligament reconstruction with Intrafix]. AB - OBJECTIVE: To investigate the clinical results of anterior cruciate ligament (ACL) reconstruction with Intrafix. METHODS: From March to December in 2005, there were 35 patients with ACL rupture using quadruple semitendinosus and gracillis. Endobutton was used to fix the graft at the femoral site. Thirty-two cases were followed up at the average time of 15 months (12-20 months). In 15 patients the subjective knee function evaluation, KT-2000 and X ray were performed, and MRI was performed in 11 cases either. Other 17 cases had only the subjective knee function score. RESULTS: The average IKDC score was 91 (83-97). In 15 cases there were 10 cases whose knee examination results of IKDC were A grade, 5 cases B grade (because of limited range of motion). Lysholm scores were average 89 (83-93), 17 cases excellent and 15 cases good. The results of KT-2000 were average 1.2 mm (0-2.0 mm) at 30 degree 133 N, average 0.5 mm (-0.5-2.0 mm) at 90 degree 133 N. The appearance and tension of reconstructed ACL were good on MRI. The X ray showed good position of bone tunnel. CONCLUSION: ACL reconstruction with Intrafix can restore the stability of knee, and the clinical result is good. PMID- 18509965 TI - [Arthroscopically assisted radiofrequency probe to treat achilles tendinitis]. AB - OBJECTIVE: To evaluate the effectiveness of micro-tenotomy using a radiofrequency (RF) probe to treat chronicity achilles tendinitis. METHODS: Seventeen cases of chronicity achilles tendinitis were treated by RF probe. Eleven cases were male, and 6 female. The average age of the patients was 25 (17-48) years. Nine were athlete, 4 sports activity and 4 students. Seven were in left side, and 10 right side. Before receiving the RF therapy all patients had achilles tendinitis symptoms. The patients were treated with local anesthesia under arthroscopy by Arthrocare 2000 and TOPAZ. The operation were performed through a artificial lacuna under the subcutaneous tissue of achilles tend, insert arthroscope. The pathology test found the achilles tendon surface fibrous tissue hyperplasia and tear. The probe of RF to perforate just as meshwork, the space was 3-5 mm. RESULTS: Patients reported significantly reduced pain from 7 to 14 d postoperatively. The symptoms of pain was completely disappeared in 15, obviously relieve in 2. The functional outcome was assessed using the VAS score evaluation, perioperative 8.7 and postoperative 1.6. There were no perioperative and postoperative complications related to the procedure, as rupture of achilles tendon, blood vessel and nerve injury. No infection and recur was found in the cases. CONCLUSIONS: RF therapy for chronicity achilles tendinitis under the arthroscopy with minimum invasion is less pain and easy for early rehabilitation. The result is satisfactory. PMID- 18509966 TI - [Combined segmental resection osteotomy with dual axial rotation correction, fixation and fusion for the treatment of severe angular kyphosis]. AB - OBJECTIVE: To design a new surgical correction and fixation technique for the treatment of severe angular kyphosis, observe the feasibility, safety and effectiveness of the technique. METHODS: From May 2004 to February 2007, 16 cases with severe kyphosis (average 90.8 degrees, range 50 degrees-130 degrees) were treated with segmental resection osteotomy, section distraction, dual axial rotation correction and instrumentation fusion technique. The patients were inspected by local and total spine anteroposterior and lateral radiography pre and postoperatively. The kyphotic Cobb angle was measured and 7 cases combined scoliosis Cobb angle was also measured. The Frankel Grading for neurological function, Oswestry Disability Index (ODI) and Patients Satisfactory Index (PSI) were evaluated preoperatively, postoperatively and at follow-up. The back pain relief was also observed. RESULTS: The average surgical duration was 6.9 hours. The average blood loss was 4000 ml. The complications include 1 shifting of artificial vertebrae, 3 nerve root injury, 3 dural tear and 1 transitory dysfunction of lower extremity. All of these complications were relieved greatly after feasible treatment. The average follow-up time was 25 months. The average kyphotic angle was 90.8 degrees preoperatively, which was improved to 26.9 degrees immediately after surgery, and got an average correction rate of 72.5%. At follow-up, the average kyphotic angle was 28.9 degrees, and correction rate was 70.1%. The 7 cases who combined with scoliosis had an average Cobb angle of 35.9 degrees preoperatively, which decreased to 4.4 degrees immediately after surgery, and the correction rate was 87.2%. The correction rate was kept until follow-up (78.6%). Some patients got an improved neurological function. The Frankel Grading were E in 5 cases, D in 5 cases, C in 5 cases, and B in 1 case preoperatively. There were 10 cases of E grade, 3 cases of D grade, and 3 cases of C grade at follow-up. Except 3 cases who had no symptoms before surgery, the average ODI was 18.9 preoperatively, and 10.8 postoperatively. The average improvement of ODI was 52.7%. The PSI result showed a satisfied rate of 93.8%. The back pain of 3 cases were totally relieved after surgery. CONCLUSIONS: Segmental resection osteotomy with dual axial rotation correction and fusion technique is an effective way to treat severe angular kyphosis. It is a safe technique and has high correction rate. The long-term results is acceptable. PMID- 18509967 TI - [Analysis of the parameters about selective anterior fusion of thoracolumbar/lumbar curves in adolescent idiopathic scoliosis]. AB - OBJECTIVE: To establish criteria for AIS of Lenke5 and Lenke6 by an anterior only procedure of the lower curve fusion. METHODS: A retrospective study was conducted between March 1999 and May 2004 to investigate 52 AIS patients of Lenke5 and Lenke6. All the patients were observed 24 years (34 months on average). Many parameters were evaluated. At final assessment, two groups emerged: Group A had satisfactory results (the thoracic curve was reduced) and Group B had just the opposite. RESULTS: Preoperative thoracic curve in group A averaged 33 degrees and 18 degrees after surgery. The lumbar curve averaged 49 degrees before surgery and 21 degrees after surgery. In group B (n = 6), the average thoracic curve was 38 degrees before surgery and 45 degrees after surgery, whereas the lumbar curve averaged 46 degrees before surgery and 25 degrees after surgery. Two of these patients underwent posterior thoracic instrumentation and fusion because of the unreasonable balance. CONCLUSIONS: A successful surgical outcome was dependent on both the flexibility of the thoracic curve and the patients' maturity. The thoracolumbar/lumbar-thoracic (TL/L:T) Cobb ratio in combination with the flexibility of the thoracic curve were the best predictors among the structural indexes. PMID- 18509968 TI - [Comparative percutaneous with open pedicle screw fixation in the treatment of thoracolumbar burst fractures without neurological deficit]. AB - OBJECTIVE: To compare clinical outcome of the percutaneous versus open pedicle screw fixation in the treatment of thoracolumbar burst fracture with neurological intact. METHODS: Sixty patients with thoracolumbar burst fracture without neurological deficit underwent either percutaneous (n = 30) or traditional open pedicle screw fixation (n = 30). Radiographs obtained before surgery, immediately after surgery, 4 months and 2 years after surgery were used to access the restoration of spinal anatomy. Also, operation time, blood loss, blood drainage, hospital stay and soft tissue dissection were evaluated. The level of pain was assessed by visual analog scale (VAS), function by the Oswestry questionnaire. RESULTS: The average followed up was 2 years. There were no significant differences between both groups concerning age, sex, cause of injury and the presence of other severe injuries. Significant differences were observed between the two groups in blood loss, blood drainage, hospital stay and soft tissue dissection (P < 0.01), whereas no significant differences in operation time (P > 0.05). The vertebral height, the kyphosis angle, and the occupation of spinal canal after surgery and at follow-up were not significantly (P > 0.05). The pain systems and functions were similar in both groups at final follow-up (P > 0.05), however, less pain was found in the percutaneous group than that in the open group at the first 3 months after surgery (P < 0.01). CONCLUSION: Percutaneous pedicle screw fixation for thoracolumbar fracture has the advantage of less trauma, quickly recovery and better esthetic outcome, however, it has the same results with the traditional open produce after 2 years of surgery. PMID- 18509969 TI - [Analysis of veracity of the C1 lateral mass screw insertion in the atlantoaxial fixation]. AB - OBJECTIVE: To evaluate the veracity of the C1 lateral mass screw insertion in the atlantoaxial fixation using plate and screw method without A-P fluoroscopic guiding. METHODS: In the atlantoaxial fixation, without A-P fluoroscopic guiding, we probe lateral mass edge and identify the entrance point of C1 lateral mass screw indirectly. 159 patients treated with atlantoaxial fixation using plate and screw method were studied. Postoperative CT was used for analysis, and the position of the lateral mass screws in atlas was identified by coronary and axial CT scan. Three areas were delimited in and around the lateral mass of atlas in the axial CT scan, so as to analyze the location of screws: area A (inside the joint face), area B (outside the joint face but still in lateral mass), and area C (outside the lateral mass). RESULTS: Among the 318 screws, 308 (96.9%) were located in area A, 5 in area B (1.6%), and 5 in area C (1.6%). All cases got atlantoaxial union at 4 months after operation. CONCLUSION: Probing lateral mass edge and identifying the lateral mass indirectly is reliable for identifying the path of screw in atlas. PMID- 18509970 TI - [Effectiveness of preoperative autologous plateletpheresis combined with intraoperative autotransfusion on the blood coagulation in orthopaedic patients]. AB - OBJECTIVE: To investigate the effectiveness of preoperative plateletpheresis combined with intraoperative autotransfusion on the blood coagulation of orthopaedic patients. METHODS: Sixty patients (ASA I-II) undergoing selective orthopaedic surgery were randomized into three groups (n = 20), that is, preoperative plateletpheresis combined with intraoperative autotransfusion for group I, intraoperative autotransfusion for group II, and group III without any managements of blood conservation. Coagulation parameters (prothrombin time, partial thromboplastin time, fibrinogen), hemoglobin and hematocrit values, platelet counts and aggregability were evaluated before the anaesthesia, 10 minutes after plateletpheresis, 10 minutes before the infusion of platelet rich plasma or autologous blood, 10 minutes after infusion, 24 and 48 hours postoperation. Intra- and postoperation blood loss and homologous blood transfusion requirements were also recorded. RESULTS: Among three groups, there were no differences in intraoperative blood loss, perioperative haemoglobin level (Hb and Hct). As compared with group I, significant lower level of platelet counts and aggregability were observed in group II and III at the time of 24 and 48 hours after operation (P < 0.05), while postoperation blood loss and homologous blood-transfusion requirements increased at the same period (P < 0.01). CONCLUSIONS: Preoperative plateletpheresis combined with intraoperative autotransfusion can ameliorate the blood coagulation in orthopaedic patients, and it is an effective way to decrease blood loss and homologous blood-transfusions requirements. PMID- 18509971 TI - [Prophylactic use of antibiotics in selective colorectal operation: a randomized controlled trial]. AB - OBJECTIVE: To investigate the reasonable proposal of prophylactic antibiotics use in selective colorectal operation. METHODS: One hundred and sixty-five patients underwent colorectal surgery were randomized to Treatment 1 (55 cases), Treatment 2 (50 cases) and Control (60 cases) group. The Treatment 1 group was given oral MgSO4 solution at the night before operation, and Cefradine 2.0 g (I.V.) during the induction of anesthesia, continued with tow times of intravenous Cefradine 2.0 g and 0.5% Metronidazole 100 ml at an interval of 12 hours in 24 hours after the operation. The Treatment 2 group was given the same treatment as Treatment 1, but the antibiotics would not be withdrawn until 3-5 d after operation. On the basis of the treatment of Treatment 2 group, the Control group was given oral antibiotics 2-3 days before operation. Postoperative complications including surgical site infection, stoma leakage, dysbacteriosis, and WBC, body temperature, days of hospitalization and antibiotic expenses in the three groups were observed and compared. RESULTS: There was no significant differences in surgical site infection, stoma leakage, WBC counting and its change, body temperature and hospital stay among the three groups (P > 0.05). The incidence rate of dysbacteriosis in Control group was significantly higher than that in Treatment 1 group (P < 0.05). The antibiotic expenses in the Treatment 1 group was significantly lower than those of the other two groups (P < 0.05). CONCLUSIONS: Prophylactic antibiotic use during the induction of anesthesia and 24 hours after operation was reasonable in selective colorectal operation, it can prevent the surgical site infection effectively with good social-economic effects and fewer side effects. PMID- 18509972 TI - [Long-term outcome of surgical angioplasty of left main coronary artery]. AB - OBJECTIVE: To assess long-term outcome of coronary artery diseased patients undergone the surgical angioplasty of left main coronary artery. METHODS: From September 1983 to December 2004, 162 patients were operated on for left main coronary artery stenosis with surgical angioplasty. The data were retrospectively analyzed. Operative death associated factors were evaluated with univariate analysis, and long-term survival was estimated by Kaplan-Meier analysis. RESULTS: The in-hospital mortality was 8% for all the patients but only 5.1% for the patients with simple left main coronary artery stenosis. A significant drop of operative mortality was noted in the later era of treatment (1983 to 1994 vs. 1994 to 2004). The operative death was more likely associated with no-isolated left main coronary artery pathologic feature and the urgency of surgery. An average of 102 (8 to 264) months' follow-up was completed in 95% of discharged patients. Coronary event occurred in 42 patients in the follow-up period and it was mortal for 9 cases. The overall long-term survival was 81% at 10 years and 52% at 20 years. The coronary event-free survival was 77% at 10 years and 41% at 20 years. CONCLUSIONS: Surgical angioplasty of left main coronary artery could produce excellent long-term outcome with acceptable per-operative mortality in left main coronary artery stenosis patients. This technique should deserve an important place in therapeutic options for this cohort of patients. PMID- 18509973 TI - [Experimental study of prostheses modified by three dimensions porous Ti combined bone morphogenetic proteins]. AB - OBJECTIVE: To observe bone ingrowth of artificial femur which three dimensions (3 D) porous Ti combined bone morphogenetic proteins (BMPs) integrating on the prostheses surface in dogs. METHODS: The prostheses integrated 3-D porous Ti on the surface, which combined BMPs directly or through FG, were implanted canine. And fluorescent labeling was done at 2, 5 weeks after that, and then the prostheses with femurs were taken out in 3, 6 weeks after operation. These specimens were treated, then observed through microscopy. RESULTS: At 3 weeks, bone growing 1/2 of full thickness in 3-D porous Ti, but bone growing full thickness in 3-D porous Ti at 6 weeks. Bone formation was obviously higher at 6 weeks than at 3 weeks. CONCLUSION: The prostheses modified 3-D porous Ti can accelerate osteogenesis and improve bone formation so that mechanical interlock and integration can be come true. PMID- 18509974 TI - [The effect of cyclooxygenase-2 on lymphangiogenesis in breast cancer]. AB - OBJECTIVE: To study the effect of cyclooxygenase-2 (COX-2) on lymphangiogenesis in breast cancer. METHODS: By the means of immunohistochemistry, COX-2, vascular endothelial growth factor-C (VEGF-C) and D2-40 were examined in the tissue samples of primary tumors from 94 patients underwent surgical resections for breast cancer from November 1998 to March 2002. Eighty-three patients were followed-up. The expressions of VEGF-C mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in MDA-MB-231 cell lines by the treatment of selective COX-2 inhibitor Nimesulide at different doses. The expressions of VEGF-C protein were evaluated in MDA-MB-231 cells treated by PGE2 (1 microg/ml) and Trastuzumab (1 microg/ml), respectively. RESULTS: COX-2 over-expression was observed in 46.8% of surgical specimens (44/94), while VEGF-C overexpression occurred in 51.1% of tumor samples (48/94). COX-2 was strongly correlated with VEGF-C expression (P < 0.01), micro-lymphatic vessels (P = 0.032) and metastatic lymph nodes (P = 0. 035). Patients with COX-2 positive tumors had a significant shorter survival time than those with negative tumors did, including disease-free survival (P = 0.010) and overall survival (P = 0.040). Nimesulide could down-regulate the expressions of VEGF-C mRNA and protein in a does-dependent manner, while PGE2 could up-regulate the expressions. The expression of VEGF-C protein up-regulated by PGE2 treatment was decreased by Trastuzumab. CONCLUSIONS: COX-2 over-expression can up-regulate the expression of VEGF-C. VEGF-C might promote lymph node metastasis by a lymph-angiogenic pathway, then affect the prognosis of the patients with breast cancer. PMID- 18509975 TI - [Effects of gene transfer CTLA4-Ig and anti-CD154 monoclonal antibody on the rejection of rat islet xenografts]. AB - OBJECTIVE: To study the effects of gene transfer cytotoxic T lymphocyte associated antigen 4 immunoglobulin (CTLA4-Ig) and anti-cluster of differentiation 154 (CD154) mAb on the rejection of rat islet xenografts. METHODS: Human islets were infected with the recombinant adenoviruses containing CTLA4-Ig gene. Transduced islets were transplanted under the left kidney capsule of diabetic rats. And then the animal model were treated with anti-CD154 monoclonal antibody. The changes of blood sugar were measured and the survival rates of grafts and transplantation rats were observed after transplantation. The morphological changes of grafts were observed. Expression of CTLA4-Ig and insulin were detected by immunohistochemical staining and cytokines were quantified by ELISA. RESULTS: (1) The blood glucose of transplantation rats decreased to normal level on 2nd day post-transplantation. The average level blood glucose of control group A, anti-CD154mAb treatment group B, transfected group C and associated treatment group D increased on day 8, 18, 25, 36, post-transplantation respectively. (2) The grafts of group A, B, C and D survived for (10.0 +/- 2.1) d, (22.0 +/- 8.2) d, (28.0 +/- 6.5) d and (37.0 +/- 9.3) d respectively. The survival of grafts in group D was significant longer than that in group A, B and C, respectively; The survival of group B and C were significantly prolonged compared with group A and the survival of group B was significantly different with group C (P < 0.05). The survival of transplantation rats were (21.0 +/- 5.7) d, (35.0 +/- 6.5) d, (48.0 +/- 8.5) d and (65.0 +/- 12.5) d in group A, B, C and D, respectively. The survival of transplantation rats compared each other among four groups were same as the survival of grafts (P < 0.05). (3) In control animals (group A), serum IL-2 and TNF-alpha concentration were elevated to a high level within seven days post-transplantation and significantly increased compared with that before transplantation (P < 0.01). (4) Hematoxylin-eosin staining of grafts showed a lot of islets under the kidney capsule of transplantation rats, no inflammatory cell infiltrate and immunohistochemical staining of grafts demonstrated expression of insulin protein at islets in group B, C and D. These grafts positively stained for CTLA4-Ig in group C and D. CONCLUSIONS: Gene transfer CTLA4-Ig and anti-CD154mAb treatment can inhibit the rejection of rat islet xenografts and treatment Ad-CTLA4-Ig and anti-CD154 mAb could induce immune tolerance of islet xenografts. PMID- 18509976 TI - [WHO classification of tumors of central nervous system (2007): an introduction]. PMID- 18509977 TI - [Clinicopathologic study of neuronal and mixed neuronal-glial tumors]. PMID- 18509978 TI - [Advanced molecular pathogenesis and treatment of Alzheimer's disease]. PMID- 18509979 TI - [Research on distribution and expression of NMDA receptors and parvalbumin positive neurons in intractable epilepsy-related focal cortical dysplasia]. AB - OBJECTIVE: To evaluate the alteration of subunits composition in NMDA receptor and the alterations of the expression and distribution of NMDA receptors and parvalbumin (PV)-positive neurons in focal cortical dysplasia (FCD) cortices. METHODS: Twenty cases of FCD samples (including all four subtypes of FCD) obtained during epilepsy surgery and 4 controls were analysed by immunohistochemical staining for NR1, NR2A/B and PV. RESULTS: Increased expression of NR1 was detected in the giant neurons and dysmorphic neurons in FCD; while pronounced expression of NR2A/B was detected in immature neurons, giant neurons and dysmorphic neurons of FCD, especially in somata and processes of the immature neurons. Compared with the controls, FCD cortices showed prominent scattered arrangement of PV positive neurons and fibers, dramatically decreased number of PV positive interneurons and PV background staining, especially in foci of FCD II subtype. CONCLUSION: There are increased expressions of NR1 and NR2A/B subunits in FCD abnormal neurons, as well as scattered and reduced expressions of PV positive neurons and fibers in FCD cortices. PMID- 18509980 TI - [Expression of drug resistance-associated proteins in brain of patients with refractory epilepsy]. AB - OBJECTIVE: To study the expression of P-glycoprotein, multi-drug resistance associated protein and major vault protein in pathologic brain specimens, and to investigate their roles in the pathogenesis of refractory epilepsy. METHODS: Immunohistochemical study was performed in pathology specimens from 18 cases of refractory epilepsy (including 5 cases of focal cortical dysplasia, 3 cases of tuberous sclerosis, 5 cases of ganglioglioma and 5 cases of dysembryoplastic neuroepithelial tumor). RESULTS: Both the P-glycoprotein and major vault protein were localized in microvascular endothelium of the lesions. Major vault protein was also seen in balloon cells and some neuronal cells. On the other hand, multi drug resistance associated protein was mainly localized in the neuronal component of the lesions. In general, the expression of P-glycoprotein and major vault protein in tumoral tissue was higher than that in non-tumoral tissue. The expression of multi-drug resistance associated protein and major vault protein was also different in the neoplastic glial cells of ganglioglioma and dysembryoplastic neuroepithelial tumor. CONCLUSIONS: P-glycoprotein, multi-drug resistance associated protein and major vault protein contribute to the pathogenesis of refractory epilepsy. They may however have different roles, with different cellular localization. PMID- 18509981 TI - [Extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma: a clinicopathologic and immunophenotype analysis of 5 cases]. AB - OBJECTIVE: To study the clinicopathologic features, diagnosis and differential diagnosis of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma. METHODS: Five cases of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma were selected from 102 cases of myeloid sarcoma diagnosed during the period from 1990 to 2006. The clinicopathologic findings and followup data were retrospectively analyzed. Immunohistochemical study was also carried out with SP method. RESULTS: Among the 5 cases studied, 3 were males and 2 were females, including 2 children and 3 adults. Generalized lymphadenopathy was found in 4 patients and skin lesions were observed in 2 patients. The tumor cells in all cases were positive for CD68 (KP1), CD68 (PGM1), lysozyme and CD45. They were negative for MPO, CD15, CD163, TdT, CD117, T and B cell markers. The Ki 67 index ranged from 40% to 80%. Follow-up data were available in all the 5 patients. Four of the 5 patients died of the disease, with the average survival time being 6.25 months. CONCLUSIONS: Monoblastic sarcoma is a rare disease with poor prognosis. It is almost impossible to distinguish monoblastic sarcoma from granulocytic sarcoma and other types of small round cell tumors on the basis of morphologic examination alone. Immunohistochemistry is mandatory for a correct diagnosis. PMID- 18509982 TI - [Clinicopathological features of the mucocele-like lesions in the breast]. AB - OBJECTIVE: To study the clinical and pathological features of mucocele-like lesions in the breast. METHODS: Nine cases of mucocele-like lesions in the breast were reported for the morphological and immunohistochemical features, the differential diagnosis, and a literature review. RESULTS: All nine cases were from female patients, aged 23 to 43 years (mean 34 years), clinically presented with palpable breast masses. Grossly, the lesions were multi-cystic with colloidal appearances. Histologically, the lesions consisted of multiple cysts filled with colloid, these cysts were lined with tubular, cuboidal or columnar epithelium. There were superimposed papillary epithelial hyperplasia in three cases and atypical ductal hyperplasia in one. Extravasated mucinous lakes were seen in the stroma, but without cellular component. CONCLUSION: Mucocele-like lesions of the breast is a group of mostly benign disease, and the differential diagnosis should include mucinous carcinoma. PMID- 18509983 TI - [Expression of Twist in papillary thyroid carcinomas and its roles in differential diagnosis]. AB - OBJECTIVE: To study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL). METHODS: Fifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19). RESULTS: In PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively. CONCLUSIONS: Positive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC. PMID- 18509984 TI - [Inflammatory pseudotumor-like follicular dendritic cell tumor of spleen]. AB - OBJECTIVE: To study the clinicopathologic features of inflammatory pseudotumor like follicular dendritic cell tumor of spleen. METHODS: One case of inflammatory pseudotumor-like follicular dendritic cell tumor of spleen was examined macroscopically and microscopically. Immunohistochemical study for CD21, CD23, CD35, clusterin, S-100 protein, vimentin, smooth muscle actin, CD1a, CD68, ALK protein, CD30, CD31, CD34, CD3 and CD20 was performed on formalin-fixed, paraffin embedded sections by standard EnVision method. In-situ hybridization for Epstein Barr virus (EBV)-encoded RNA was also carried out. RESULTS: Macroscopically, inflammatory pseudotumor-like follicular dendritic cell tumor was large in size, tan-colored, soft to rubbery in consistance and associated with central hemorrhage and necrosis. Histological examination showed scattered follicular dendritic cells admixed with abundant lymphocytes and plasma cells in the background, simulating inflammatory pseudotumor. On high-power magnification, the follicular dendritic cells possessed a moderate amount of pale to lightly eosinophilic cytoplasm, with indistinct cell borders. The nuclei were ovoid or spindly, with vesicular or stippled chromatin and small distinct, often centrally located, nucleoli. Some of the tumor cells showed nuclear pleomorphism and contained irregular foldings of nuclear membrane, coarse chromatin and prominent eosinophilic nucleoli. Mitotic figures were rarely identified. Immunohistochemical study showed that the tumor cells were positive for vimentin, clusterin, smooth muscle actin and CD68. They were weakly and focally positive for CD35 and S-100 protein, but negative for CD21, CD23, CD1a, ALK protein, CD30, CD31 and CD34. Most of the background lymphocytes were of T-lineage (CD3 positive) ,some were CD20 (B-cell marker)-positive. EBV RNA was demonstrated in the tumor cells by in-situ hybridization analysis. CONCLUSIONS: Inflammatory pseudotumor-like follicular dendritic cell tumor is a rarely encountered low grade malignancy with distinctive morphologic pattern. It is associated with EBV infection. PMID- 18509985 TI - [Vasculogenic potential of endothelial progenitor cells derived from human umbilical cord blood and their roles in neovascularization of malignant glioma]. AB - OBJECTIVE: To investigate vasculogenic potential of endothelial progenitor cells (EPCs) derived from human umbilical cord blood and their contribution to the neovascularization of malignant glioma in vivo. METHODS: EPCs were isolated from human umbilical cord blood by density gradient centrifugation. After 7-10 days of culture, EPCs were investigated for CD34 and VEGFR-2 expression by direct immunofluoresent staining. The proliferative activity, migratory capability and forming capillary-like tubules were also monitored after stimulation with VEGF(50 mg/L) in vitro. Moreover, EPCs were administered into tumor-bearing mice, and the tumor and mouse organs were examined under confocal laser scanning microscope to visualize the distribution and localization of transplanted EPCs. In order to quantity the incorporation of EPCs into tumor vessels, cryosections of the tumor tissue were double-labelled with antihuman CD31 and anti-mouse CD31. RESULTS: After 7 to 10 days of culture, EPCs assumed cobblestone-like monolayer growth pattern with nearly complete confluence, and expressed CD34 and VEGFR-2. Significant proliferative activity, increased migratory capability and forming capillary-like tubules were observed when stimulated with VEGF. The transplanted EPCs in vivo specifically homed to solid tumor tissue and incorporated into the tumor's endothelium. Quantitative analysis revealed that human EPCs contributed significantly to tumor neovascularization by incorporation into tumor vasculature (18.68 +/- 1.32)% of the total vessels. CONCLUSION: EPCs possess the potential to form neovascular network in tumor and play a role in the phenotypical heterogeneity of tumor microvascular architecture. PMID- 18509986 TI - [Neuroprotective effects of alpha7 neuronal acetylcholine receptor and its roles in the pathogenesis of Alzheimer's disease]. AB - OBJECTIVES: To investigate the neuroprotective function of alpha7 nicotinic receptor (nAChR) and its roles in the pathogenesis of Alzheimer's disease (AD). METHOD: Specific RNA interference to alpha7 nAChR mRNA expression was performed by gene specific small interference RNA (siRNA). SH-SY5Y cells were transfected with the siRNA or treated with 20 micromol/L 3-[2, 4-dimethoxybenzylidene] anabaseine (DMXB), an alpha7 nAChR agonist. After 48 hrs culture, levels of alpha7 nAChR mRNA and protein were monitored by RT-PCR and Western blotting, respectively. In the second experiment, SH-SYSY cells treated with siRNA or DMXB were exposed to 1 micromol/L Abeta(25-35), followed by protein analysis of alpha form of secreted beta-amyloid precursor peptide (alphaAPPs), and total APP was assayed by Western blotting. In addition, lipid peroxidation and MTT [3-(4,5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] reduction were measured by spectrophotometry. RESULT: In RNA interference group, as compared with controls, alpha7 nAChR mRNA and protein levels were decreased with inhibitory efficiency by 80% and 69%, respectively, along with a decrease in protein levels of alphaAPP and reduction of MTT. However the product of lipid peroxidation was increased. There was an enhanced gene inhibition of alpha7 nAChR by Abeta. While cells treated with DMXB, the alpha7 nAChR protein was increased by 23% as compared with that of the control, along with decrease of alphaAPP and ERK 1/2 at the protein level. The enhanced expression of alpha7 nAChR reduced the neurotoxic effects resulted from Abeta. CONCLUSION: The findings indicate that alpha7 nAChR may play a significant neuroprotective role by enhancing cleavage of APP, improving antioxidant defenses and limiting the toxicity of Abeta, which has been implied in the pathogenesis of AD. PMID- 18509987 TI - [Expression of cancer stem cell antigens, prostate stem cell antigen and Oct-4, and its clinicopatholgical significances in benign and malignant lesions of gallbladder]. PMID- 18509988 TI - [Relationship between bone marrow microenvironment and tumor metastasis]. PMID- 18509989 TI - [Applications of adhibiting agent on tooth and bone sections]. PMID- 18509990 TI - [Liponeurocytoma of right lateral ventricle: report of a case]. PMID- 18509991 TI - [Atypical meningioma of left lateral sulcus with chondroid metaplasia: report of a case]. PMID- 18509992 TI - [Malignant histiocytosis-like B-cell lymphoma of liver: report of a case]. PMID- 18509993 TI - [Survey of the perioperative prophylactic use of antimicrobial agents in 118 hospitals in China]. AB - OBJECTIVE: To investigate and analyze the perioperative prophylactic use of antimicrobial agents in 118 hospitals in China. METHODS: 3557 medical records (from September to December, 2006) of 118 hospitals were drawn out randomly. The perioperative prophylactic use of antimicrobial agents was investigated and analyzed. RESULTS: Prophylactic antimicrobial agents were used in 3485 cases (98%). The first 3 kinds of antimicrobial agents most in use were cephalosporins of 3rd generation (1775/3485, 50.4%), 2nd generation (1191/3485, 34.2%) and fluoroquinolones (1120/3485, 34.1%). The average durations of antibiotic use were 7.4 d for class I (clean) wounds 7.6 d for class II (clean/contaminated) wounds and 10. 5 d for class III (contaminated) wounds. Only 30.4% of patients received antibiotics within 2 h prior operation, and 52.2% of patients did not received antibiotics until the operation was completed. There were no indications for prophylactic antibiotic use in 16.7% of cases. Combining use of antimicrobial agents were performed in 56.5% of cases, and 22.1% of them lack of reasonable indications. CONCLUSIONS: Inappropriate use of perioperative prophylactic antimicrobial agents is common and must be subjected to standardization. PMID- 18509994 TI - [Surgical resection of hepatic tumor in segment IXb by anterior transhepatic approach]. AB - OBJECTIVE: To evaluate the feasibility and the effect of surgical resection of hepatic tumor originated from segment IXb. METHODS: The cases with hepatic tumors in segment IXb who had been operated on between March 2003 and January 2007 were retrospectively reviewed. RESULTS: A total of 15 tumors in segment IXb, including 13 primary liver cancers and 2 benign tumors with a mean diameter of (4.3 +/- 1.6) cm, were successfully resected by anterior transhepatic approach under sequential occlusions of portal tride and total hepatic vascularity or portal tride clamping only. There was no operative mortality,with a mean operative time of (190.3 +/- 37.6) min and a mean operative blood lose of (376.7 +/- 252.7) ml. All the patients had uneventful postoperative course except one who suffered from ascites and edema of the low body, which was successfully managed medically. The mean postoperative hospital stay was (13.3 +/- 6.0) d. During the follow-up of 1 47 months, two patients with benign tumor enjoyed a normal life. Among the 13 patients with primary live cancers, 1 patient died of recurrence, 2 patients remained alive with intrahepatic recurrence and 10 patients survived without any sign of relapse, with a median tumor-free survival time of 23.5 months. CONCLUSIONS: Surgical resection of hepatic tumor in segment IXb, despite their sophisticated anatomic position, is feasible in technique with high safety. The local resection can provide the patients with potential to cure. PMID- 18509995 TI - [Analysis of risk factors for selective devascularization in patients with portal hypertension]. AB - OBJECTIVE: To investigate the risk factors for selective devascularization in patients with portal hypertension. METHODS: The clinical data of 160 patients with portal hypertension underwent selective devascularization were retrospectively analyzed. All the patients were divided into high-risk group and low-risk group according to the postoperative complications. Thirty-two clinical factors were analyzed using logistic regression. RESULTS: Single-factor analysis showed that history of jaundice, Child-Turcotte-Pugh classification, total bilirubin (before the operation), prolongation of prothrombin time, pre-operative free portal pressure, ascites, leukocyte count (1 week after the operation) and hemoglobin (1 week after the operation) were significantly different between the high-risk group and low-risk group (P < 0.05). Logistic regression analysis showed that decrease of free portal pressure, total bilirubin (before the operation), prolongation of prothrombin time, ascites, leukocyte count (1 week after the operation) and hemoglobin (1 week after the operation) were still significantly different between the two groups (chi2 = 53.337, P < 0.01). CONCLUSIONS: The risk factors of selective devascularization in patients with portal hypertension are decrease of free portal pressure, pre-operative total bilirubin, prolongation of prothrombin time, ascites, post-operative leukocyte count and hemoglobin. PMID- 18509996 TI - [Laparoscopy-assisted D2 total gastrectomy in advanced gastric cancer]. AB - OBJECTIVE: To investigate the efficacy and advantages of laparoscopy-assisted total gastrectomy (LATG) with D2 dissection of lymph nodes versus conventional open D2 total gastrectomy (OTG) in advanced gastric cancer. METHODS: One hundred and twenty-five patients with advanced gastric cancer in the middle or upper third of the stomach were operated on from July 2005 to March 2007. Of the patients, 59 cases received LATG and 66 OTG with D2 lymph nodes dissection. Clinical data were recorded and compared between the two groups. RESULTS: No patient in the LATG group converted to conventional operation with laparotomy. No operation mortality and no severe morbidity occurred in LATG group. As compared with OTG group, in LATG group operation time was longer [(330 +/- 71) min vs. (261 +/- 54) min, P =0.005] in LATG group, but with similar number of lymph node retrieval (36 +/- 13 vs. 34 +/- 16, P =0.450), less operation blood loss [(175 +/ 101) ml vs. (359 +/- 210) ml, P =0.003], earlier recovery of bowel activity (P = 0.015), and a shorter duration of fever after operation (P = 0.024). CONCLUSIONS: LATG with D2 lymph node dissection in advanced gastric cancer is safe and technically feasible with better operative access and visual field, less operation blood loss and earlier recovery. PMID- 18509997 TI - [The utility of stapler in distal pancreatectomy]. AB - OBJECTIVE: To determine the influence of hand-sewn and stapler in distal pancreatectomy on the postoperative complication. METHODS: Clinical data of 109 patients after distal pancreatectomy from January 2003 to December 2006 were analyzed retrospectively. RESULTS: The surgical techniques used for closure of the pancreatic stump after distal pancreatectomy were categorized into hand-sewn closure group (n = 53) and stapler closure group (n = 56). In stapler closure group, 25 patients accepted laparoscopic operation. The incidences of abdominal infection and pancreatic fistulae in stapler closure group were lower than hand sewn closure group. The operation time, blood infusion, postoperative bleeding and medical costs were similar between two groups. CONCLUSION: Stapler closure in distal pancreatectomy could decrease the incidence of pancreatic fistula and abdominal infections. PMID- 18509998 TI - [Study on the three dimensional hepatic virtual operation based on the data of 64 slice helical CT scanning]. AB - OBJECTIVE: To study the surgery plan and simulation effect of the three dimensional (3D) hepatic virtual operation based on the data of 64-slice helical CT scanning and to probe the feasibility of the virtual operation based on the FreeForm Modeling System. METHODS: The volunteer liver was scanned to collect two dimensional (2D) DICOM data of 64-slice helical CT scanning and the 3D hepatic and intrahepatic vessels model were reconstructed by MIMICS software. The reconstructed liver, the intrahepatic vessels model and the artificial tumor models were output into the FreeForm Modeling System in the STL format. The device PHANTOM with the characterization of dynamo-feedback was applied to make the operation on the 3D hepatic. RESULTS: The spatial relationship between the tumour and the intrahepatic vessels were clearly observed by rotation and enlargement of the target. According to the operation principle, the left lobe of liver resection was simulated by manipulating the device PHANToM. Through the liver transparence surface, the intrahepatic vessels were easily distinguished. The operation procedure was accord with the clinic hepatic surgery. Meanwhile, during the operation, by adjusting the incision objective intensity, the dynamo feedback intensity was definitely touched. CONCLUSIONS: By using the FreeForm Modeling System,the hepatic operation procedure can be simulated ahead of time. The operation complication in the practical surgery can be anticipated and the individualization operation schema can be reasonable instituted. PMID- 18509999 TI - [Determination of painful vertebrae treated by kyphoplasty in multiple-level vertebral compression fractures]. AB - OBJECTIVE: To explore how to determine painful vertebrae treated by kyphoplasty in multiple-level osteoporotic vertebral compression fractures and clinical outcome. METHODS: From October 2002 to June 2005, 51 consecutive procedures with kyphoplasty were performed on 35 patients with multiple-level osteoporotic vertebral compression fractures. There were 51 painful vertebrae among 120 vertebral compression fractures. The painful vertebra was determined by the signal intensity changes in MR images, combined with radiography and local percussion pain before operation. Only painful vertebrae were treated by kyphoplasty. Preoperative, postoperative and final follow-up visual analog scale (VAS) and radiographic findings such as vertebral height and Cobb angle were recorded and analyzed. RESULTS: All patients tolerated the procedure well with immediate relief of their back pain after kyphoplasty and they can walk at 1-3 days after the procedure. There were 3 vertebrae (3/51) occurred asymptomatic extravasation of cement. 31 cases were followed up for mean 16.2 months (range 6 44 months). VAS reduced from preoperative 8.7 to final follow-up 2.1 (P <0. 01). At final follow-up the vertebral height had a recovery rate of 59.17%, and the mean Cobb angle was improved 10.1 degrees. There was a significant improvement between preoperative and final follow-up values (P < 0.01). CONCLUSIONS: The painful vertebra can be determined by signal intensity changes in MR series images in multiple-level osteoporotic vertebral compression fractures. Selecting painful vertebrae to be treated by kyphoplasty can make patients with multiple level VCFs gain an excellent result. PMID- 18510000 TI - [Effect on wrist joint stability following distal radial fractures]. AB - OBJECTIVES: To investigate the influence of wrist joint stability of patients with diversified fracture of the distal radius, and to find out the difference of the clinical effect between the patients with surgical treatment and non-surgical treatment. METHODS: From January 1999 to September 2006 a total of 200 cases with the fracture of the distal radius were reviewed according to the AO classification, the radiolunate angle, scapholunate angle, palmar tilt angle and the length of the radial shorting and step-off of the articular surface were measured by the standard X-ray. Gartland and Werley as modified by Sarmiento evaluation system was used, and the results were statistically analyzed. RESULTS: The average follow-up period was 5 years and 2 months. The follow-ups revealed 5 types of carpal instability: scapholunate dissociation, volar intercalated segment instability, dorsal intercalated segment instability and palmar or dorsal shift of the carpus. Functional results were excellent and good in 78% of the total patients. The increasing of the length of the radial shorting and step-off of the articular surface was found to be associated with greater risk of carpal instability. CONCLUSIONS: Based on the results of the study, wrist articular surface and radial shortening and palmar tilt angle should be considered as the most important factors of the healing effect after the fracture of the distal radius. Wrist joint stability depends on the structure of the bone and ligament around wrist joint. Carpal instability wound leads to significantly effect on the wrist. And surgery was necessary to severe intra-articular fracture of the distal radius. PMID- 18510001 TI - [Treatment of old femoral neck and nonunion with free vascularized fibular grafting]. AB - OBJECTIVE: To evaluate the clinical result of the treatment of old femoral neck fracture and nonunion with free vascularized fibular grafting. METHODS: From November 2000 to December 2005, 29 cases with old femoral neck fracture and nonunion had been treated by free vascularized fibular grafting with an average follow-up of 28.5 months. RESULTS: All the fracture were healed without any severe complications. And the healing time was 4-6 months (5.6 months on average). During the follow-up, the hips of 28 cases got well-function, and the average Harris hip score was 88.2. One case came about with osteonecrosis of femoral head after one year and finally accepted THA after 2 years. CONCLUSIONS: The free vascularized fibular grafting is a valuable procedure to treat old femoral neck fracture and nonunion. PMID- 18510002 TI - [The initial approach of two sides hypogastric artery ligation and isolation with laparoscope before excision of presacral tumor]. AB - OBJECTIVE: To observe the significance of excision of presacral tumor after two sides hypogastric artery ligation and tissue dissociation with laparoscope. METHODS: Twenty-one patients with sacral tumor were performed excision of presacral tumor after two sides hypogastric artery ligation and tissue dissociation with laparoscope. RESULTS: All sacral tumor were removed successfully, the mean volume of operative blood was 800 ml (range 500-1900 ml), and all the patients were followed up 3-25 months, averaged time 11 months. One patient was recurred after 2 months of operation (the patient was Ewing's sarcoma, and refused to accept radiotherapy and chemotherapy after operation), 1 patents died of brain metastases after 9 months of operations. There were no recurrence in the others patients. CONCLUSIONS: The excision of presacral tumor after two sides hypogastric artery ligation and tissue dissociation with laparoscope is an effective operation method, with the advantages of decreasing the operative blood and difficulty of sacral tumor excision, and diminishing the operation wound. PMID- 18510003 TI - [Relative factors of recent discovered atrial fibrillation following isolated coronary artery bypass grafting]. AB - OBJECTIVE: To identify the relative factors of recent discovered atrial fibrillation (AF) following isolated coronary artery bypass grafting (CABG). METHODS: Classified the 649 cases undergoing isolated CABG from January 2005 to December 2006 to two groups according to whether AF appeared after operation. Collected the peri-operative data and operative strategy, then analyzed with single-factor analysis and Logistic regression. RESULTS: The incidence of AF was 8.0% (52 cases), and 84.6% (44 cases) recovered sinus-rhythm leaving hospital. Age, standard European system for cardiac operative risk evaluation (EuroSCORE), ratio of high-operative-risk, left atrium diameter and ratio of left coronary artery dominance were higher in AF group than in non-AF group. Age, eject fraction, left atrium diameter, operative risk evaluation, left coronary artery dominance and anastomosis on right coronary artery were the relative factors of recent discovered AF following isolated CABG. But off-pump operation, prescription of adrenergic beta-antagonists pre-operatively and degree of coronary artery stenosis had no influence to AF. CONCLUSIONS: AF following CABG is a result of common influence by many factors. EuroSCORE might forecast partially the incidence of AF following CABG. Improve the myocardial protection and reduce the surgical damage during operative progress maybe the mostly approach to decrease the incidence of AF following CABG. PMID- 18510004 TI - [The clinical analysis of 54 cases for the surgical treatment of pulmonary embolism]. AB - OBJECTIVE: To evaluate the results of surgical procedures for pulmonary embolism. METHODS: Fifty-four patients of pulmonary embolism received surgical treatment from October 1994 to June 2007, of which 9 were acute pulmonary embolism underwent pulmonary embolectomy and 45 patients were chronic thromboembolic pulmonary hypertension (CTEPH) underwent pulmonary thromboendarterectomy. RESULTS: The mortality rate was 44.4% in acute pulmonary embolism group and 13.3% in CTEPH group (P < 0. 05). Thirteen patients had residual pulmonary hypertension and 23 patients had severe pulmonary reperfusion injury postoperatively. The pulmonary artery systolic pressure changed from (89.4 +/- 36.3) mm Hg (1 mm Hg =0.133 kPa) preoperative to (55.6 +/- 22.4) mm Hg postoperative. The pulmonary vascular resistance changed from (89. 7 +/- 56.7) kPa L(-1) S(-1) preoperative to (38.9 +/- 31.1) kPa L(-1) S(-1) postoperative. The arterial partial pressure of oxygen changed from (52. 3 +/- 6.7 ) mm Hg preoperative to (87.6 +/- 6.5) mm Hg postoperative. The arterial oxygen saturation changed from (88.9 +/- 4.5)% preoperative to (95.3 +/- 2.8 )% postoperative (P < 0.05). With the follow-up of (41.8 +/- 36.4) months, there were 4 patients died. According to NYHA, there were 28 patients for class I , 10 patients for class II and 2 patients for class III. According to Kaplan-Meier survival curve, the 3-year, 4-year, 5-year and 8-year survival rate were (97.1 +/- 2.8 )%, (94.0 +/- 4.1)%, (90.8 +/- 5.2)% and (85.0 +/- 7.3)% respectively. Linear rate of bleeding and thromboembolic related to anticoagulation were 0. 63% patient-years and 0. 62% patient-years respectively. CONCLUSIONS: The operational mortality of acute pulmonary embolism is significantly higher than CTEPH, and the mid-long term survival rate is agreeable and the complication rate related to anticoagulation is relatively low. PMID- 18510005 TI - [Relationship between the intrapelvic perfusion pressure in minimally invasive percutaneous nephrolithotomy and postoperative recovery]. AB - OBJECTIVES: To introduce the method to monitor intrapelvic perfusion pressure during minimally invasive percutaneous nephrolithotomy (MPCNL), and to observe the effect of high pressure intrapelvic perfusion on recovery. METHODS: The end of F5 ureteral catheter and aseptic transducer were connected by self-made connecter. During the operation, 46 renal calculi cases were monitored, and the early complication, such as fever, pain index, drop of hemoglobin, the stone-free rate and hospital stay were investigated. And the relationship between the variation of pressure and recovery was studied. RESULTS: Intrapelvic perfusion pressure ranged from 3 mm Hg (1 mm Hg = 0.133 kPa) to 50 mm Hg during the course of MPCNL. The definition of high pressure was the time of the pressure more than 30 mm Hg not less than 10 min. Postoperative fever rate, pain index, drop of hemoglobin and hospital stay in the high pressure were significantly higher than low pressure group (P< or =0.05). There was no difference of the stone-free rate in two groups. CONCLUSIONS: During the course of MPCNL intrapelvic perfusion pressure should be monitor immediately. It should be careful to maintain the time of pressure more than 30 mm Hg less than 10 min for stable postoperative recovery. PMID- 18510006 TI - [70 degrees recumbent position transperitoneal laparoscopy for treatment of upper urinary tract transitional cell carcinoma]. AB - OBJECTIVE: To study the effect of 70 degrees recumbent position transperitoneal laparoscopy for treatment of upper urinary tract transitional cell carcinoma (TCC). METHODS: From May 2004 to January 2007, 70 degrees recumbent position transperitoneal laparoscopy combined with urethral resectoscope was used to treat 31 cases of upper urinary tract transitional cell carcinoma. At the same time titanium clip to occlude the two extremities of ureter tumor was used, extracting specimen by oblique incision of lower quadrant. RESULTS: All operations were finished successfully, no one was turned to open surgery; mean operation time was 140 min, mean blood loss 80 ml, mean hospital stay time 8 d, without complications of urine leakage and intestinal fistula and so on. CONCLUSIONS: 70 degrees recumbent position transperitoneal laparoscopy for resection of whole kidney and ureter is worth of general clinical application because it could provide large space for operation, simplify the treatment of renal pedicle vessels, decrease operation risk, reduce operation trauma and offer early recovery. But its effect on tumor spread and recurrence will still need long term follow-up. PMID- 18510007 TI - [Prevention of facial nerve injury in acoustic neuroma microsurgery]. AB - OBJECTIVE: To summarize and analyse the techniques of avoiding facial nerve injury during acoustic neuroma microsurgery. METHODS: One hundred and eighty patients with large acoustic neuroma (> or =4 cm) and 70 patients with medium acoustic neuroma (2.4-4.0 cm) were diagnosed by MRI/ CT scan before operation and confirmed by postoperative pathologic examination. All of patients were treated by sub-occipital retrosigmoid approach for tumor removal and facial nerve reservation during operation. The relationships among the bone, arachnoid, nerve and vascular anatomy were particularly observed during the operation. After decompression of the tumor, the origination and location of the facial nerve as well as the relationship between the tumor and the facial nerve should be identified. The patients were followed-up from 6 months to 1 year postoperatively and assessed by House-Brackmann facial nerve function grading system. RESULTS: Total tumor resection was achieved in 240 of 250 cases (96%) and subtotal in 10 cases including 1 case died because of cerebellar encephalomalacia after operation. According to the House-Brackmann facial nerve function grading, recovery of normal function (grade I) was achieved in 214 cases (85.6%), grade II in 25 cases (10%), grade III in 5 cases (2.09%) and grade IV in 5 cases (2.09%). CONCLUSION: Microneurosurgical techniques are helpful for total resection of acoustic neuroma and keeping facial nerve anatomic intact. PMID- 18510008 TI - [Inhibitory effects of ODC and AdoMetDC bi-antisense virus on the growth and invasion of lung cancer cell A-549]. AB - OBJECTIVE: To study the inhibitory effects of antisense bicistronic recombinant adenovirus vector of ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (Ad-ODC-AdoMetDCas) on polyamine biosynthesis,proliferation and invasion of lung cancer cells. METHODS: Adenovirus-mediated gene transduction efficiency was assessed with counting GFP-positive cells using trypan blue. Western Blot and HPLC were used to detect ODC and S-AdoMetDC expression and polyamine content in A-549 cells respectively. Viable cell counting and cell cycle analysis were adopted to evaluate cell growth and cell cycle distribution, and A-549 cell invasion in vitro was detected with Matrigel invasion assay. RESULTS: Approximate 75% of A-549 cells were infected with Ad-ODC-AdoMetDCas when multiplicity of infection reached 50. Our study demonstrated that Ad-ODC AdoMetDCas vector-mediated gene transfer inhibited tumor cell growth through the blockade of polyamine synthesis pathway. The tumor cells were arrested at cell cycle G1 phase after gene transfer. Gene transferred tumor cells were shown to possess markedly decreased invasiveness. CONCLUSION: Ad-ODC-AdoMetDCas has significant inhibitory effects on lung cancer cell proliferation and invasion and bears therapeutic potential for the treatment of lung cancer. PMID- 18510009 TI - Transgenesis and the management of vector-borne disease. Preface. PMID- 18510010 TI - Perspectives on the state of insect transgenics. AB - Genetic transformation is a critical component to the fundamental genetic analysis of insect species and holds great promise for establishing strains that improve population control and behavior for practical application. This is especially so for insects that are disease vectors, many of which are currently subject to genomic sequence analysis, and intensive population control measures that must be improved for better efficacy and cost-effectiveness. Transposon mediated germ-line transformation has been the ultimate goal for most fundamental and practical studies, and impressive strides have been made in recent development of transgene vector and marker systems for several mosquito species. This has resulted in rapid advances in functional genomic sequence analysis and new strategies for biological control based on conditional lethality. Importantly, advances have also been made in our ability to use these systems more effectively in terms of enhanced stability and targeting to specific genomic loci. Nevertheless, not all insects are currently amenable to germ-line transformation techniques, and thus advances in transient somatic expression and paratransgenesis have also been critical, if not preferable for some applications. Of particular importance is how this technology will be used for practical application. Early ideas for population replacement of indigenous pests with innocuous transgenic siblings by transposon-vector spread, may require reevaluation in terms of our current knowledge of the behavior of transposons currently available for transformation. The effective implementation of any control program using released transgenics, will also benefit from broadening the perspective of these control measures as being more mainstream than exotic. PMID- 18510011 TI - Alphavirus transducing systems. AB - Alphavirus transducing systems (ATSs) are important tools for expressing genes of interest (GOI) in mosquitoes and nonvector insects. ATSs are derived from infectious cDNA clones of mosquito-borne RNA viruses (family Togaviridae). The most common ATSs in use are derived from Sindbis viruses; however, ATSs have been derived from other alphaviruses as well. ATSs generate viruses with genomes that contain GOI's that can be expressed from additional viral subgenomic promoters. ATSs in which an exogenous gene sequence is positioned 5' to the viral structural genes is used for stable protein expression in insects. ATSs in which a gene sequence is positioned 3' to the structural genes is used to trigger RNAi and silence expression of that gene in the insect. ATSs are proving to be invaluable tools for understanding vector-pathogen interactions, vector competence, and other components of vector-pathogen amplification and maintenance cycles in nature. These virus-based expression systems also facilitate the researcher's ability to decide which gene-based disease control strategies merit a further investment in time and resources in transgenic mosquitoes. PMID- 18510012 TI - Paratransgenesis applied for control of tsetse transmitted sleeping sickness. AB - African trypanosomiasis (sleeping sickness) is a major cause of morbidity and mortality in Subsaharan Africa for human and animal health. In the absence of effective vaccines and efficacious drugs, vector control is an alternative intervention tool to break the disease cycle. This chapter describes the vectorial and symbiotic biology of tsetse with emphasis on the current knowledge on tsetse symbiont genomics and functional biology, and tsetse's trypanosome transmission capability. The ability to culture one of tsetse's commensal symbiotic microbes, Sodalis in vitro has allowed for the development of a genetic transformation system for this organism. Tsetse can be repopulated with the modified Sodalis symbiont, which can express foreign gene products (an approach we refer to as paratransgenic expression system). Expanding knowledge on tsetse immunity effectors, on genomics of tsetse symbionts and on tsetse's parasite transmission biology stands to enhance the development and potential application of paratransgenesis as a new vector-control strategy. We describe the hallmarks of the paratransgenic transformation technology where the modified symbionts expressing trypanocidal compounds can be used to manipulate host functions and lead to the control of trypanosomiasis by blocking trypanosome transmission in the tsetse vector. PMID- 18510013 TI - Bacteria of the genus Asaia: a potential paratransgenic weapon against malaria. AB - Symbiotic bacteria have been proposed as tools for control of insect-borne diseases. Primary requirements for such symbionts are dominance, prevalence and stability within the insect body. Most of the bacterial symbionts described to date in Anopheles mosquitoes, the vector of malaria in humans, have lacked these features. We describe an alpha-Proteobacterium of the genus Asaia, which stably associates with several Anopheles species and dominates within the body of An. stephensi. Asaia exhibits all the required ecological characteristics making it the best candidate, available to date, for the development ofa paratransgenic approach for manipulation of mosquito vector competence. Key features of Asaia are: (i) dominance within the mosquito-associated microflora, as shown by clone prevalence in 16S rRNA gene libraries and quantitative real time Polymerase Chain Reaction (qRT-PCR); (ii) cultivability in cell-free media; (iii) ease of transformation with foreign DNA and iv) wide distribution in the larvae and adult mosquito body, as revealed by transmission electron microscopy, and in situ hybridization experiments. Using a green fluorescent protein (GFP)-tagged Asaia strain, it has been possible to show that it effectively colonizes all mosquito body organs necessary for malaria parasite development and transmission, including female gut and salivary glands. Asaia was also found to massively colonize the larval gut and the male reproductive system of adult mosquitoes. Moreover, mating experiments showed an additional key feature necessary for symbiotic control, the high transmission potential of the symbiont to progeny by multiple mechanisms. Asaia is capable of horizontal infection through an oral route during feeding both in preadult and adult stages and through a venereal pattern during mating in adults. Furthermore, Asaia is vertically transmitted from mother to progeny indicating that it could quickly spread in natural mosquito populations. PMID- 18510014 TI - Proposed uses of transposons in insect and medical biotechnology. AB - Transposons are small pieces of DNA that can transpose through either RNA or DNA intermediates. They have been found in almost all organisms and are important components of the evolutionary process at the chromosomal level. They have provided the raw genetic material that has produced domesticated genes that now provide important cellular functions and are now being explored as genetic tools in both humans and insects that vector human pathogens. Here I compare the requirements for both insect and human gene therapy and discuss the similarities between them in terms of transposon performance. Recent progress in understanding transposon function in terms of transposase structure is described as is the rapidly emerging role of RNAi in generic transposon regulation. These developments reinforce the view that, autonomous, transposon behavior in host organisms is, in part, determined by the nuclear and cellular environment of the cell and these factors need to be considered when developing transposons as therapeutic agents either in humans or in insects that vector human disease. PMID- 18510015 TI - The Yin and Yang of linkage disequilibrium: mapping of genes and nucleotides conferring insecticide resistance in insect disease vectors. AB - Genetic technologies developed in the last 20 years have lead to novel and exciting methods to identify genes and specific nucleotides within genes that control phenotypes in field collected organisms. In this review we define and explain two of these methods: linkage disequilibrium (LD) mapping and quantitative trait nucleotide (QTN) mapping. The power to detect valid genotype phenotype associations with LD or QTN mapping depends critically on the extent to which segregating sites in a genome assort independently. LD mapping depends on markers being in disequilibrium with the genes that condition expression of the phenotype. In contrast, QTN mapping depends critically upon most proximal loci being at equilibrium. We show that both patterns actually exist in the genome of Anapheles gambiae, the most important malaria vector in sub-Saharan Africa while segregating sites appear to be largely in equilibrium throughout the genome of Aedes aegypti, the vector of Dengue and Yellow fever flaviviruses. We discuss additional approaches that will be needed to identify genes and nucleotides that control phenotypes in field collected organisms, focusing specifically on ongoing studies of genes conferring resistance to insecticides. PMID- 18510016 TI - Impact of technological improvements on traditional control strategies. PMID- 18510017 TI - Insect population suppression using engineered insects. AB - Suppression or elimination of vector populations is a tried and tested method for reducing vector-borne disease, and a key component of integrated control programs. Genetic methods have the potential to provide new and improved methods for vector control. The required genetic technology is simpler than that required for strategies based on population replacement and is likely to be available earlier. In particular, genetic methods that enhance the Sterile Insect Technique (e.g., RIDL) are already available for some species. PMID- 18510018 TI - Wolbachia-based technologies for insect pest population control. AB - Wolbachia are a group of obligatory intracellular and maternally inherited bacteria found in many arthropod species, including insects, mites, spiders, springtails, crustaceans, as well as in certain nematodes. Several PCR-based surveys suggest that over 20% of the arthropod species may be Wolbachia-infected, rendering this bacterium the most ubiquitous intracellular symbiont yet described. Wolbachia have recently attracted attention for their potential as novel and environmentally friendly bio-control agents. Wolbachia are able to invade and maintain themselves in the arthropod species through manipulation of the host's reproduction. Several strategies can be distinguished, one of which is cytoplasmic incompatibility (CI). Wolbachia-induced cytoplasmic incompatibility can be used beneficially in the following ways: (a) as a tool for insect pest population control in a way analogous to the "Sterile Insect technique" (SIT) and (b) as a drive system to spread desirable genotypes in field arthropod populations. In addition, virulent Wolbachia strains offer the potential to control vector species by modifying their population age structure. In the present chapter, I summarize the recent developments in Wolbachia research with an emphasis on the applied biology of Wolbachia and conclude with the challenges that Wolbachia researchers will face if they want to use and/or introduce Wolbachia into pest and vector species ofeconomic, environmental and public health relevance and, through Wolbachia-based technologies, to suppress or modify natural populations. PMID- 18510019 TI - Using predictive models to optimize Wolbachia-based strategies for vector-borne disease control. AB - The development of resistance to insecticides by vector arthropods, the evolution of resistance to chemotherapeutic agents by parasites and the lack of clinical cures or vaccines for many diseases has stimulated a high-profile effort to develop vector-borne disease control strategies based on release of genetically modified mosquitoes. Because transgenic insects are likely to be less fit than their wild-type counterparts, transgenic traits must be actively driven into the population in spite of fitness costs (population replacement). Wolbachia are maternally-inherited symbionts that are associated with numerous alterations in host reproductive biology. By a variety of mechanisms, Wolbachia-infected females have a reproductive advantage relative to uninfected females, allowing infection to spread rapidly through host populations to high frequency in spite of fitness costs. In theory, Wolbachia can be exploited to drive costly transgenes into vector populations for disease control. Before conducting an actual release, it is important to be able to predict how released Wolbachia infections are expected to behave. While inferences can be made by observing the dynamics of naturally occurring infections, there is no ideal way to empirically test the efficacy ofa Wolbachia gene driver under field conditions prior to the first actual release. Mathematical models are a powerful way to predict the outcomes of transgenic insect releases and allow one to identify knowledge gaps, identify parameters that are critical to the success of releases, conduct risk-assessment analysis and investigate worst-case scenarios, and ultimately identify the most effective, most logistically feasible control method or methods. In this chapter, I review current and historical advances in applied models of Wolbachia spread, specifically within the context of applied population replacement strategies for vector-borne disease control. PMID- 18510020 TI - Modifying insect population age structure to control vector-borne disease. AB - Age is a critical determinant of the ability of most arthropod vectors to transmit a range of human pathogens. This is due to the fact that most pathogens require a period of extrinsic incubation in the arthropod host before pathogen transmission can occur. This developmental period for the pathogen often comprises a significant proportion of the expected lifespan of the vector. As such, only a small proportion of the population that is oldest contributes to pathogen transmission. Given this, strategies that target vector age would be expected to obtain the most significant reductions in the capacity of a vector population to transmit disease. The recent identification of biological agents that shorten vector lifespan, such as Wolbachia, entomopathogenic fungi and densoviruses, offer new tools for the control of vector-borne diseases. Evaluation of the efficacy of these strategies under field conditions will be possible due to recent advances in insect age-grading techniques. Implementation of all of these strategies will require extensive field evaluation and consideration of the selective pressures that reductions in vector longevity may induce on both vector and pathogen. PMID- 18510021 TI - Technological advances to enhance agricultural pest management. AB - Biotechnology offers new solutions to existing and future pest problems in agriculture including, for the first time, possible tools to use against insect transmitted pathogens causing plant diseases. Here, we describe the strategy first described as Autocidal Biological Control applied for the development of conditional lethal pink bollworm strains. When these strains are mass-reared, the lethal gene expression is suppressed by a tetracycline repressor element, which is activated by the presence of chlorotetracycline, a normal component of the mass-rearing diet. Once removed from the tetracycline diet, the lethal genes are passed on to offspring when ordinary lab-reared pink bollworms mate with special lethal strains. Lethality is dominant (one copy sufficient for lethality), expressed in the egg stage and affects all eggs (100% lethal expression). The initial investment by the California Cotton Pest Control Board is an outstanding example of research partnerships between agriculture industry, the USDA and land grant universities. PMID- 18510022 TI - Applications of mosquito ecology for successful insect transgenesis-based disease prevention programs. PMID- 18510023 TI - A rational look at the apparent mess of bariatric surgery procedures and indications. PMID- 18510024 TI - [The management of obstructive jaundice in pancreatic cancer]. AB - Patients with pancreatic cancer often present with advanced disease; so, curative surgical resection is possible in a small number of patients. Palliation in these patients focuses particularly on relief of biliary obstruction. Palliative treatment modalities include both surgical and nonsurgical approaches. Biliary obstruction is initially treated with endoscopic biliary stenting, plastic or metallic stents. Both of these provide similar initial relief of biliary obstruction; however, plastic stents have a greater risk of occlusion and should be used in patients with short survival duration. Metallic stents have a greater initial cost, but provide an overall cost-saving in patients with expected survival more than 6 months. There is no evidence of benefit from routine stenting of jaundiced patients before resection. Surgical palliation for biliary obstruction should be primarily considered in patients who fail endoscopic or percutaneous biliary decompression or who develop gastroduodenal obstruction, It is also indicated for patients with good performance status and expected survival of over 6 months. Surgical decompression of biliary tree should be made with a choledochojejunostomy whenever feasible, associated to a gastroduodenal bypass. PMID- 18510025 TI - [The acute cholecystitis: the operative timing for the laparoscopic approach]. AB - The aim of this study is to evaluate the results of early cholecystectomy in patients with acute cholecystitis. In the past, acute cholecystitis was a contraindication to laparoscopic cholecystectomy because of the greater risk of injury to the biliary duct, but acute gallbladder inflammation was a contraindication to open cholecystectomy, too. With greater experience and new technology, laparoscopic cholecystectomy is today the gold standard in the treatment of acute cholecystitis, in empyema and gangrenous cholecystitis. In recent years, attention has turned to surgical timing, rather than surgical management--open versus laparoscopy--because there is no advantage in delaying cholecystectomy for acute cholecystitis. In our experience, we always choose laparoscopic technique in all the patients without general contraindications to mini-invasive surgery and operate as soon as possible in a patient with unfavourable conditions. We believe that the patient must be quickly stabilized with preoperative medical procedures, and surgical treatment must be performed within 72-96 hours after the onset of symptoms. During this period, laparoscopic approach allows a reduction of operative time, operative risk and the conversion rate with medical and economic advantages. PMID- 18510026 TI - [Postoperative follow-up of gastric adenocarcinoma with neoplastic markers and 18 FDG-PET/TC]. AB - INTRODUCTION: The usefulness of tumour markers CEA, CA19.9 and CA72.4 in association with FDG-PET/TC were prospectively evaluated in the post-operative follow-up of gastric cancer patients. MATERIAL AND METHODS: Fifty one consecutive patients were enrolled in a follow-up programme entailing with periodical clinical evaluations, instrumental examinations and tumour markers assay FDG PET/TC was performed only in cases of suspected recurrence. RESULTS: Sensitivity of CEA, CA19.9 e CA72.4 during the follow-up period was respectively: 16%, 33.3% e 50%. Overall sensitivity was 66.6%. Specificity was 100% for CEA, 93.3% for CA19.9, 100% for CA72.4, with an overall specificity of 96.2%. FDG-PET/TC had a sensitivity of 100%. CONCLUSIONS: Tumour markers in association with FDG-PET/TC allow an early identification of recurrences after surgery, with the advantage to start chemotherapy or surgical protocols before the tumour has reached an advanced stage. PMID- 18510027 TI - [Our experience in selecting patients for bariatric surgery]. AB - AIM OF THE STUDY: To evaluate the parameters applied in Authors' experience for the selection of candidates undergoing bariatric surgery. MATERIALS AND METHODS: A retrospective analysis has been performed of 447 patients who underwent bariatric surgery from December 1998 to December 2006 at the 1st Service of General Surgery of Spedali Civili of Brescia - Abdominal Surgical Department of University of Study of Brescia; 317 patients underwent a mal-absorptive procedure, 35 a gastro-restrictive procedure and 10 had a conversion from a gastro-restrictive procedure to a mal-absorptive one. RESULTS: At 7 years of follow-up the Authors observed that the excess weight loss was 74.1% +/- 6.2 for mal-absorptive procedures and 42.3% +/- 2.6 for gastro-restrictive procedures. At 2 years of follow-up a progressive weight resumption was observed in the 20% of patients who had a gastro-restrictive procedures. DISCUSSION AND CONCLUSIONS: Optimal clinical outcome were achieved confirming the belief that a strict and rigorous selection of the patients and the applied therapeutic algorithm have to be followed; though even more selective criteria can be suggested such as the preventive BIB. The clinical results supported our preference of mal-absorptive procedures (317 of 352 surgical performances, excepting the 10 conversions with an obliged surgical choice). PMID- 18510028 TI - [The role of neoadjuvant radio-chemotherapy in the treatment of rectal cancer]. AB - OBJECTIVE: To evaluate oncological and surgical outcome of patients submitted to neoadjuvant therapy for advanced rectal cancer. PATIENTS AND METHOD: One hundred thirty eight patients (86 male, 52 female, mean age 61.4 years), with tumour of lower (58; 42%), middle (66; 48%), upper rectum (14; 10%), showing a clinical stage II (23; 17%) or III (115; 83%) and with an average distance from anal verge of 6.5 cm, submitted to fractionated "long-course" RT with CT locally staged by US and MR before and after neoadjuvant therapy and operated on after 4-6 weeks by its end. RESULTS: Surgical procedures (71 of which laparoscopic) were: 114 AR (83.8%), 19 APR (14%) and 3 TEM (2.2%). Mean nodal-sampling was 14.9. A complete or partial response was observed in 48.5% of the patients (67/138). With a mean follow-up of 30 months, local recurrence rate was 5.7%. Five-years overall survival and disease-fee-survival were respectively 73% and 60%. DISCUSSION: We observed a significant clinical (p < 0.004) and pathological (p < 0.005) downstaging. Pre-treatment clinical stage was not significant. On the contrary, postoperative yTNM was significant for yT (p < 0.001) and yN (p < 0.0003). Non responder patients had worse prognosis (5-years survival 30%). The variable with higher prognostic significance was yN (p < 0.0003), especially if we distinguish N1 by N2 (p < 0.0004). CONCLUSIONS: The response to neoadjuvant therapy represents a significant prognostic variable. PMID- 18510029 TI - [Correlations between splenectomy and allergic symptoms. Preliminary results of a study concerning IL-4 and IgE serum level]. AB - The Authors, studying the condition of immunodepression observed in patients who underwent splenectomy for traumatic lesion, noticed that a significant part of them complained of allergic symptoms. These seemed to appear only in subjects whose Th2 lymphocytes functionality was preserved--as witnessed by normal or increased IL-4 serum level--showing an increased level of IgE too, thus confirming the role of Th2 lymphocytes in stimulating IgE synthesis. On the contrary splenectomized patients with functional harm of the Th2 lymphocytes- proved by low IL-4 serum level--did not show increase of haematic IgE level nor allergic manifestations. The Authors stress that, despite the small numerical of the series not allowing definitive judgment, preliminary data are suggestive for an hypothesis--reported below--that needs further confirmation. The ablation of the spleen, organ devoted to remove from the blood antigens--many of these potentially allergic factors--, allows the prolonged persistence of such sensitizing agents, thus promoting the onset of allergic manifestations in splenectomized patients whose Th2 lymphocytes function is preserved. PMID- 18510030 TI - The superior pedicle mammaplasty for the treatment of pedunculous breast. AB - AIM OF THE STUDY: The superior pedicle mammaplasty is a technique frequently employed in the treatment of breast ptosis, associated or not with hypertrophy of the gland, followed by satisfactory results. This technique is normally not indicated in severe breast ptosis (pendulous breast) because of the excessive length of the pedicle supplying nipple-areola-complex (NAC), with the risk of ischemia. In these cases the standard technique is the free-nipple-graft mammaplasty. However, the deepen knowledge about vascular anatomy of the breast and the aptitude to perform superior pedicle mammaplasty, induced the authors to indicate this technique even in these cases improving the aesthetic and functional outcomes. MATERIAL AND METHODS: The authors present a series of 30 patient with pendulous breasts, with sternal notch-nipple distance equal or superior to 32 cm (45 cm maximum; mean value 35.1), treated with the superior pedicle mammaplasty with inverted "T" scar. The results confirm the reliability of superior pedicle for the nipple-areolar complex blood supply, associated with satisfactory aesthetic results due especially to the good breast projection. CONCLUSIONS: They conclude that superior pedicle technique mammaplasty, even if normally not indicated in these cases, is instead suitable for the treatment of pendulous breasts with great sternal notch-nipple distance, permitting to take advantages of this technique. PMID- 18510031 TI - Squamous cell carcinoma of the breast. Report of two cases. AB - Squamous Cell Carcinoma (SCC) of the breast is a rare entity, not well known, nevertheless literature reports several signalings from different authors. Regarding ethiopathogenesis, SCC of the breast is still the object of numerous discordances and controversies. We report two cases of SCC of the breast referred to our institution in the last year. The first case interests a 35 years-old woman with a lesion of the left breast referred to us with a 3 month history of breast mass. The second case regards a 49 year-old women with multicentric lesions of the right breast operated in two different times. The SCC is a rare breast entity with a controversial histogenesis. The development is characterized by metastasis frequency. The treatment remains not codified. The treatment is currently the same as for infiltrating ductal carcinoma. Indeed, the limited number of this kind of tumor doesn't allow the assessment of different therapeutic protocols. PMID- 18510032 TI - The management of acute gastric volvulus. AB - Acute gastric volvulus occurs when the stomach or a part of it rotates more than 180 degrees which leads to obstruction (closed loop syndrome), that finally concludes to incarceration and ischemia of the organ. It can be observed as a result of diaphragmatic hernia, a gap of the diaphragm, pancreatic or gastric cancers, traumatic injuries and fixation anomalies. Pari first described acute gastric volvulus in 1579, in a patient with diaphragmatic injury after trauma. The first reports of successful surgical repair and anatomo-pathologic findings were published in 1866 and 1897. PMID- 18510033 TI - [Giant retroperitoneal tumors: a 13.5 kg liposarcoma case report]. AB - Retroperitoneal sarcomas (RS) are a rare group of malignant soft-tissue tumors. They generally grow until they reach large size before becoming symptomatic, often involving surrounding structures. CASE REPORT: The paper reports the case of a particularly large retroperitoneal liposarcoma (diameter: 48 x 44 x 32 cm, weight: 13.500 kg) surgically treated in a 92-year old woman: the patient tolerated the procedure well and was discharged after a short hospitalization. The paper, moreover, focuses on the diagnostic and therapeutic aspects of this tumour. The need for radical surgery to remove the entire mass is certain, even if it is particularly large; furthermore, the patient must be carefully monitored since retroperitoneal recurrence is frequent and must be treated, wherever possible, surgically. After a review of the literature there is an evidence that higher tumor grade, non-liposarcoma histology, advanced stage, incomplete surgical resection, and microscopic infiltration of surgery resection margins were found to be significantly negative prognostic factors. CONCLUSIONS: The study underlines the role of liposarcoma surgery management in order to offer the best chance of long-term survival and, especially on elderly patients, a better quality of life. PMID- 18510034 TI - [Amyand's hernia. Case report]. AB - The authors report a case of Amyand's hernia and describe the pathophysiology, the diagnosis and the therapy in occurrence of this surgical condition. PMID- 18510035 TI - Rare complication after anterior resection of the rectum and colon reconstruction: severe constipation with obstructed defecation. Report of a case. AB - OBJECTIVE: The aim of this manuscript is to verify the impact that some recanalization procedures for intestinal continuity could have in bowel function and quality of life. STUDY MATERIAL: We describe a clinical case of a rectal cancer patient who underwent anterior resection of the rectum with colo-anal anastomosis, colo-plasty and diverting ileostomy. RESULTS: After the diverting ileostomy closure, suffered of severe bowel function problems. The establishment of a colo-plasty caused a syndrome of such severe obstructed defecation to necessitate the reestablishment of a diverting ileostomy. DISCUSSION: Anterior resection with total mesorectal excision and colo-anal anastomosis is the gold standard surgical treatment of rectal carcinoma. The so called "anterior resection syndrome" is well known after such surgical procedures. The establishment of a reservoir such as the J-pouch and more recently the transverse pouch (colo-plasty) are procedures used to improve the quality of life after anterior resection of the rectum. CONCLUSIONS: The presence of bowel obstruction without mechanical causes makes us consider the colo-plasty as its cause with a Hirschprung like mechanism or similar to the "obstructed defecation". The peristalsis stops at the colo-plasty level impeding the progression of feces. The colo-plasty or the pouch do not function as a reservoir to accommodate feces, but because they stop the peristalsis. PMID- 18510036 TI - Splenic infarction, rare cause of acute abdomen, only seldom requires splenectomy. Case report and literature review. AB - Splenic infarction is a rare disorder, commonly without a characteristic symptomatology, that rarely requires surgical procedure; in fact it has the peculiar feature of an high tendency for complete healing after the only medical approach. Furthermore in order to prevent OPSI, only in case of clear, persistent symptoms or complications it may be necessary surgical laparoscopic or open approach. The Authors report on a recent case observed in emergency of splenic infarction with infectious complications that made the surgical procedure mandatory. PMID- 18510037 TI - Are porcine and human TSH receptor antibody measurements comparable? AB - BACKGROUND: The human TSH receptor antibody (TRAb) assay is reported to show higher sensitivity in comparison to the "classical" porcine TRAb testing. This claim is based on studies comparing porcine TSH receptor (TSH-R) in one type of assay system (non-immobilized TSH-R and PEG to separate bound and free) with recombinant human TSH-R in another type of assay system (TSH-R immobilized on plastic tubes). In this study isotopic TRAb assays both based on the "coated tube" system (second generation) were compared for the first time. MATERIALS AND METHODS: Three hundred and nineteen consecutive undiagnosed patients from Mainz University as well as 72 patients from Dresden University with known Graves' disease were tested in the human TRAb assay and in the porcine TRAb assay. Spearman's coefficient of rank correlation and Chi-square test statistical analysis was performed. RESULTS: In the 319 consecutive patients 35 patients (11.0%) were found positive in the human assay and 36 patients (11.3%) were positive in the porcine assay. Seventy patients with Graves' disease from Dresden were positive in the assay using the porcine TSH-R and 71 out of 72 patients showed positive results in the assay with the recombinant human TSH-R. The Spearman's coefficient of rank correlation for both patient cohorts was r = 0.932 and r = 0.891 (p < 0.001), respectively. There was no significant difference in the clinical assessment regarding the diagnosis of Graves' disease. CONCLUSION: There is no clinically relevant difference in isotopic TRAb second generation assays--independent of the TSH-R used. Both TSH-R assays showed nearly similar TRAb results in consecutive samples as well as in serum samples from patients with Graves' disease. PMID- 18510038 TI - Diagnostic value of five commercial tests for the rapid diagnosis of Clostridium difficile-associated disease. AB - We compared five commercial immunoassays (Biostar OIA CdTOX AB, ImmunoCard Toxins A&B - Meridian, Xpect C. difficile toxin A/B -Remel, C. difficile toxin A test- Oxoid, and TOX A/B QUIK CHEK- Techlab) which allow a rapid diagnosis of C. difficile associated disease. The tests were performed directly on patient's stool specimen submitted for routine investigation of C. difficile infection from two University Hospitals in Brussels. The cell cytotoxicity assay was considered as the gold standard. Of the 100 stool specimens included in the study 23 were positive for C. difficile toxin. The sensitivity, specificity, positive and negative predictive values were respectively, 95.7%, 100%, 100% and 98.7% for TOX A/B QUIK CHEKTM, 91.3%, 100%, 100% and 97.5% for ImmunoCard Toxins A&B and for Xpect C. difficile toxin A/B, 87%, 100%, 100% and 96.3% for OIA CdTOX AB and 87%, 98.7%, 97.2% and 96.3% for C. difficile toxin A test. The differences were not statistically significant (p > 0.05). These data suggest that the tested immunoassays are acceptable for rapid detection of C. difficile toxin. PMID- 18510039 TI - The knocked-out erythrocyte sedimentation rate: periodontal abscess. AB - INTRODUCTION: The erythrocyte sedimentation rate (ESR) is a common but nonspecific test that is often used as an indicator of active disease. Infection of dental origin may be responsible for a number of cases in unresolved elevated ESR and fever etiology. Dental sepsis is the one of the potential causes of persistent fever that can escape detection. CLINICAL PICTURE: An 18-year-old female patient was admitted to the emergency room with complaints of headache, fever, nausea, and vomiting for the past four days. Erythrocyte sedimentation rate was 110 mm/h. She was started empirically on antibiotic treatment as no etiology was found. Four days later, while searching for the etiology of the fever, the patient experienced an acute pain in association with localizing symptoms in two decayed teeth. Oral examination revealed abscess formation in both teeth. TREATMENT: Teeth were extracted and ESR was decreased to 95 mm/h on the day of the second extraction and to 60, 35, and 10 mm/h taken weekly. OUTCOME: During the follow-up, she was in good health with no fever seen 3 months after treatment and her ESR was 15 mm/h. CONCLUSION: Dental infection should be considered as an unusual but very treatable cause of pyrexia of unknown origin. PMID- 18510040 TI - SimChip--computer simulation of mRNA steady states. AB - We developed a public Internet program called SimChip (www.simchip.de). It performs virtual "in silico experiments", which reflect the in vivo situation of gene expression. Our computer model simulates up- and downregulation of genes on the basis of differential equations for linear synthesis and non-linear decay of mRNA. Steady-state concentrations are described as ratios of the respective rate constants: c = S/D. Using physiological synthesis rates of 0 to 50 mRNA molecules per min per cell and decay rates of 0.05 to 25% per min, SimChip helps to understand the right-skewed distribution patterns of gene expression levels and calculates the experimental endpoint of the theoretically infinite time needed to achieve perfect equilibrium. PMID- 18510041 TI - Novel system for worldwide qualified transport of red blood cell concentrates (RBC) under extreme environmental conditions. PMID- 18510043 TI - [Metformin therapy in polycystic ovary syndrome]. AB - Polycystic ovarian syndrom (PCOS) affects 5% to 10% of women of reproductive age, and is the most common reason of anovulation in infertile women. It is multi symptom disease. A drug which is used to induce ovulation in women with PCOS is clomiphene citrate. After clomiphene treatment, ovulation is achieved in 56%-73% of women. The rest of them has different level of opportunity to therapy. Clomiphene does not have influence on elementary factor in PCOS which seems to be opportunity to insulin. The drug which can have positive effect is metformin, derivative from biguanid. Its influence on percent of ovulation, pregnancy and live births was evaluated based on randomized trials. Many trials have reserched this so far, but their results are divergent. In this article, we are aiming to systemize results of randomized trials concerning the role of metformin in PCOS therapy. PMID- 18510044 TI - [The course of pregnancy and delivery in women after genetic amniocentesis before and after 35 years of age]. AB - OBJECTIVES: The demand for genetic amniocentesis in case of young pregnant women has significantly increased due to various new indications. Moreover, nowadays a growing number of women aged > or =35, who required genetic amniocentesis, get pregnant The aim of the following study has been to compare the course of the pregnancy, the delivery and the condition of the newborn in two groups of patients: 18-34 years old and > or =35 years old. MATERIAL AND METHODS: 783 women underwent the procedure of amniocentesis at the Department of Obstetrics of Medical University of Gdansk in 1996-2003. A group of 540 women, who answered the questionnaire about the course of the pregnancy and the delivery after the procedure, has been isolated. RESULTS: Fetal loss occurred in 2 cases (1.6%) in the group of the younger women and in 8 cases (0.8%) in the group of > or =35 year-olds (p=0.84). There was no statistically significant difference between younger and older patients when comparing complications after the procedure in the first three weeks following amniocentesis: spotting, bleeding, leakage of amniotic fluid. Frequency of late complications such as gestational diabetes mellitus, pregnancy induced hypertension and urinary tract infections were comparable in both age groups women. Cesarean section has been more frequently performed in case of the older women than in the group of younger patients, with statistical difference p=0.003. In most cases (33.9%) it has been an elective cesarean section, performed due to advanced maternal age rather than any obstetric cause. Frequency of pneumonia and the number of respiratory infections in the newborns have been comparable in both groups. CONCLUSIONS: Prenatal invasive diagnosis has no influence on frequency of complications during pregnancy and delivery in the group of women less than 35 years old and more than 35 years old. PMID- 18510045 TI - [Evaluation of the relationship between circulating nucleated red blood cells count and inborn infection in neonates]. AB - OBJECTIVES: to determine the relationship between the initial nucleated red blood cells (nRBC) count during the first 12 hours after birth and inborn infection in neonates. MATERIALS AND METHODS: The retrospective study comprised of 306 neonates born in the Department of Perinatology of the I Chair of Gynaecology and Obstetrics in Lodz, Poland, in the years 2002-2007, among whom the nucleated red blood cells count were calculated within the first 12 hours after birth. Two categories of nRBC count: the normal and the elevated value, were statistically elaborated by a Mann-Whitney test and a chi-square test with two clinical outcome categories: the presence and the absence of inborn infection in the analyzed neonates. Statistical significance was indicated by p value lower than 0,05. RESULTS: Among 306 newborns, there were 127 mature neonates (41.5%) and 179 prematures (58.5%). The mean of the initial nRBC count in the analyzed newborn population was 40, 15. The mean of the nRBC count in the infected neonates was three times higher (52.56) than the mean of the nRBC count in newborns without inborn infection (16.76) - (p=0.00001). Inborn neonatal infection concerned a vast majority of cases with an elevated value of the nRBC count (86.4%), but in 13.6%, inborn infection was not observed. Among the cases with a normal nRBC count, the presence and the absence of inborn infection was diagnosed in about 50% of the analyzed babies (50.83% vs 49.17%). The elevated value of the nRBC count in infected neonates concerned mainly premature babies, rather than mature neonates, and similarly in neonates with a lower Apgar score than in babies born in good condition. CONCLUSIONS: 1. The positive association between elevated initial nucleated red blood cells count after birth and inborn infection in newborns has been revealed. 2. An elevated nucleated red blood cells count may be an auxiliary, early indicator for inborn infection in neonates. 3. Prematurity and perinatal asphyxia favour the elevation of a nucleated red blood cells count in cases with inborn infection. PMID- 18510046 TI - [Diaphragmatic hernia in reference hospital ICZMP--diagnostic problems and outcome]. AB - OBJECTIVES: The aim of the study was to analyze US/ECHO examinations in fetuses with diaphragmatic hernia (DH) diagnosed and treated in our institution from 1994 2006, and their follow-up. MATERIAL AND METHODS: Retrospective analysis of the data base from Department for Diagnoses & Prevention of Fetal Malformations, Research Institute of the Polish Mother's Memorial Hospital: 14,481 fetal echo/ultrasound examinations in 10,077 fetuses have been analyzed to retrieve 115 fetuses with DH. RESULTS: The mean gestational age at the targeted US/ECHO examination was 30 wks. There were 8 terminations of pregnancies (at mean 21 wks), 6 intrauterine demises, 60 neonatal deaths after delivery (in 1-3rd day of postnatal life), 8 deaths after surgery, 19 neonates were discharged home and in 14 cases the follow-up could not be monitored. The most common anomalies accompanying DH have been central nervous system anomalies (20%), polyhydramnion (16%) and cong heart defects (10%). In this subgroup, there was 100% mortality. Isolated DH has been diagnosed in every third case. In this subgroup, 27 neonates had undergone surgery and the survival rate was 70%, however since 2004 there was not a single death on record. CONCLUSIONS: Late gestational age of US/ECHO examinations in our tertiary center suggests that DH has been relatively difficult to detect during ultrasound screening. DH and the other structural malformations have been a lethal disease in our series in 100%. Isolated DH was much less frequent and was present in every third case (29%), and in this group the survival rate was 70%, regardless of the way of the delivery (CS or Vaginal). PMID- 18510047 TI - [Expression of tumor necrosis factor-alpha (TNF-alpha) on peritoneal fluid mononuclear cells in women with endometriosis]. AB - OBJECTIVES: Tumor necrosis factor-alpha (TNF-alpha) plays a key role in the processes underlying the development of pelvic endometriosis. TNF-alpha acts on target cells via two receptors: TNFR1(p55) and TNFR2(p75). Depending on cell type and its activation state, ligand binding to TNF-alpha may induce activation and proliferation of the cells or promote apoptosis. The aim of our study has been to evaluate the expression of TNFR1 and TNFR2 on peritoneal fluid macrophages and T lymphocytes derived from women with endometriosis. MATERIAL AND METHODS: The study group consisted of 22 patients with endometriosis (stage I and II rAFS). 14 patients with benign, non-inflammatory ovarian tumors composed the reference group. Mononuclear cells have been isolated from peritoneal fluid, obtained during laparoscopy. The expression of TNFR1 and TNFR2 proteins has been evaluated by means of flow cytometry, using monoclonal antibodies against CD120a, CD120b, CD3 and CD14. RESULTS: The percentage of peritoneal fluid macrophages revealing the expression of TNFR1 and TNFR2 proteins has been higher in patients with endometriosis, in comparison with control group (22.6+/-5.3% vs. 6.8+/-1,8%; p=0.03 and 29.3+/-2.3% vs. 8.8+/-1.8%; p=0.01, respectively). The percentage of T lymphocytes with the expression of TNFR1 and TNFR2 has been similar in endometriosis and control group. CONCLUSION: Higher percentage of peritoneal fluid macrophages expressing TNFR1 and TNFR2 proteins in endometriosis suggests dependence of these cells on TNF-alpha stimulation. Changes in TNF receptors distribution on PF macrophages, but not lymphocyte, may play its role in the pathogenesis of endometriosis. PMID- 18510048 TI - Changing trends in the surgical treatment of female stress urinary incontinence- twenty two years observation. AB - OBJECTIVES: The aim of this study was to analyze the changing trends in surgical treatment of female urinary incontinence (UI). MATERIAL AND METHODS: Medical records of all women admitted to II Department of Gynecology from 1985 to 2006 were analyzed in order to find out how the female SUI treatment changed over these years. RESULTS: During analyzed time 36819 patients were hospitalized in our Department and 77.6% (28568) of them were operated because of various indications. The number of SUI surgeries among all hospitalized women steadily rose from 1.93% in 1985 to 10.96% in 2006 reaching maximum in 2005 (13.73%). Clinical effectiveness of SUI surgeries markedly improved from 35% for anterior colporrhaphy to almost 90 % for suburethral slings. CONCLUSIONS: Introduction into clinical practice modern suburethral slings improved clinical efficacy of SUI treatment. The percentage of women admitted and treated surgically because of SUI steadily increased over the last years. PMID- 18510049 TI - [Current views on treatment of the ovarian granulosa-cell tumor]. AB - The granulosa-cell tumor (folliculoma) is a rare type of ovarian neoplasm, accounting for 5% of all cases. It is the most common type of sex cord-stromal tumors, diagnosed in 70% of cases. The granulosa-cell tumor is a hormone active one, originating from granulosa cells which produce estradiol. Overproduction of estradiol is helpful in the diagnosis of the tumor because of its numerous symptoms. There are two types of folliculoma: juvenile (5%) and adult (95%). The juvenile type is mostly recognized (90%) in FIGO I stage and has a better prognosis. Operation is often a sufficient way of treatment in this group. Tumors in higher stages are more aggressive and must be treated further. The adult folliculoma is more aggressive in its nature. Patients with the disease diagnosed in higher stages must be treated by adjuvant radiotherapy or chemotherapy. Recurrence appears often many years after the treatment and has high mortality. Many old (platinum) and new (taxans) agents are active when used in treatment of this type of tumor. Randomized study must be made to establish standard therapy of granulosa-cell tumor. Currently, the most frequent way of treatment is chemotherapy with BEP (Blemycyna, Etopozyd, Cisplatyna). PMID- 18510050 TI - [SARS-CoV infection and pregnancy]. AB - SARS is a highly contagious infection, caused by new coronavirus SARS-CoV. Immunopathological mechanisms responsible for the reaction to SARS-CoV infection have not yet been fully elucidated. Cytokine profile of SARS patients showed marked elevation of Th1 cytokine, interferon gamma, inflammatory cytokines for at least 2 weeks after the onset of the disease. The clinical manifestation of SARS in patients has been of varied nature. Fever of more then 38 degrees C, lasting more then 24 hours, is the most frequently encountered symptom. Other symptoms are non specific and they may include: sore throat, myalgia and nausea. The results of the radiological investigation may appear normal. Infants born to pregnant women with SARS did not appear to have acquired the infection through vertical transmission. However, direct contact with the maternal body fluid which contained SARS-CoV, has put the infants in great danger of perinatal infection. Ribavirin and corticosteroids are usually suggested for the treatment of SARS. However, the ribavirin therapy increases the risk of teratogenic effects in newborns of pregnant women with SARS. Therefore, the usage of this drug is not recommended during pregnancy and lactation. PMID- 18510051 TI - [The significance of beta-3 adrenergic receptor polymorphism in excessive body gain and obesity development in pregnant women]. AB - The current knowledge allows for the definition of the genetic factors contributing to pathological body mass gain and obesity development. One of the possible candidates is the gene of beta3-adrenergic receptor (ADRB3). Due to the fact that the basic function of this receptor is the induction of thermogenesis process and an increase of energy expenditure, the significance of Trp64Arg polymorphism of ADRB3 gene in disturbances of metabolic processes, which may induce disturbances of adipocytes function and lead to excessive body mass gain, obesity and early beginning of diseases connected with obesity (dyslipidemia, obesity, chronic hypertension, diabetes mellitus type II), has been suggested. The investigations performed in pregnant women concern, in the majority of cases, women with obesity, diabetes mellitus and gestational hypertension. These studies indicate the possible correlation of higher value of body mass index (BMI) with the presence of mutated 64Arg allele. In healthy pregnant women the relationship between Trp64Arg polymorphism of ADRB3 receptor and body gain has been suggested. Additionally, in female carries of the mutated 64Arg allele, a higher placenta mass and birth mass of newborns have been noted. This publication is merely single voice in the discussion because only one cause of excessive body gain and obesity presented here - Trp64Arg polymorphism of ADRB3 receptor. The problem seems to be more complicated and requires an investigation of the influence of other genetic and environmental factors, and the future populate investigations in Trp64Arg polymorphism of ADRB3 receptor. PMID- 18510052 TI - [A rare case of post-partum urethrovaginal fistula. Management of obstetric complications]. AB - A rare case of a 22-year old patient with obstetric urethrovaginal fistula, resulting in urinary incontinence, has been reported in the following report The emphasis is put on a number of medical and social consequences related to the formation of the fistula. Authors have presented the diagnostic difficulties. The aim of the report is to draw attention to the probable complications following prolonged labour and the necessity of appropriate treatment. PMID- 18510053 TI - [Extensive vulva cancer with enormous tumor of the vagina]. AB - The following is a case report documenting a 53-year old patient with extremely advanced vulva cancer (stage IV). The patient needed urgent operative treatment because of massive bleeding from large tumor located inside and out of the vagina. The prime step was to stop tumor hemorrhage, perform cystotomy and decompression of the urinary bladder and exteriorization of artificial anus. Second operation was a conventional oncologic procedure, which contained: radical vulectomy, hysterosalpingo-oophorectomy, cystectomy with excision of the urethra, the excision of almost the entire vagina, lymphadnectomy of the left inguinal lymph nodes and iliac lymph nodes. In spite of the surgical treatment, the patient died at the fourth day after the second operation. PMID- 18510054 TI - [Avoiding common mistakes in FHR monitoring. (Part I)]. PMID- 18510055 TI - [Local use of estradiol (Vagifem) in urogynecology--the statement of Polish Gynecological Society experts]. PMID- 18510056 TI - Estimation of the relative liver perfusion using two methods of radionuclide angiography in the patients with hemodynamic disorders in the portal system. AB - The aim of this study is the assessment of the relative arterial and venous contribution to the total liver blood flow (hepatic perfusion index-HPI), with two methods (S1 and S2), and estimation of their value. With this correction, HPI nonsignificantly increases (p>0.05) in all the groups of patients, with a very high correlation between the HPI (S1) and HPI (S2) values (p<0.01). In comparison to the portal perfusion in controls, values were significantly (p<0.01) lower in chronic active hepatitis and liver cirrhosis and differed between themselves (p<0.01). In the groups of cirrhotic patients with esophageal varices, sclerosated esophageal varices, recanalized umbilical vein, portal thrombosis and cavernous portal vein, portal perfusion was lower (p<0.01) than in controls, chronic active hepatitis and liver cirrhosis without collaterals. Both angioscintigraphic methods are useful for the estimation of the disturbances in the portal system. Because of the more exact estimation of the liver perfusion, S2 is recommended. PMID- 18510057 TI - [Occurrence and risk factor for development of pancreatitis and asymptomatic hyperamylasemia following endoscopic retrograde cholangiopancreatography--our experiences]. AB - INTRODUCTION: Chronic pancreatitis is defined as chronic inflammatory lesion of pancreatic parenchyma leading to destruction and fibrosis of exocrine pancreas. Endoscopic retrograde cholangiopancreatography (ERCP) is the most sensitive and specific method for detection of morphological alterations in chronic pancreatitis. ERCP is inevitably associated to post-ERCP acute pancreatitis, as well as hyperamilasemia. STUDY AIM: This study aims to determine frequency of post-ERPC pancreatitis and asymptomatic hyperamilasemia. STUDY METHODS: We have studied 160 patients who underwent ERCP in Institute of Digestive Diseases, Clinical Centre of Serbia in Belgrade. Data regarding cholecystectomy, papillotomy, peripapillary diverticulosis, Oddi's sphincter hypertension, choledoch canulation and diameter, Wirsung duct canulation, minor duodenal papilla patency, anomalies of BP junction, as well as chronic pancreatitis has been analysed and correlated with eventual development of post-ERCP pancreatitis and asymptomatic hyperamilasemia. RESULTS: Asymptomatic hyperamilasemia was determined in 51 subjects (31.9%), while pancreatitis has been developed in 5 patients (3.1%) subsequent to ERCP. It has been proofed that Wirsung duct canulation plays significant role in development of post-ERCP complications. CONCLUSION: Although numerous factors may potentially contribute to development of post-ERCP pancreatitis, none of them, with the exception of Wirsung duct canulation, has been determined to play significant role in development of these complications. PMID- 18510058 TI - Diagnostic and therapeutic approaches to bleeding from lower parts of the digestive system. AB - Bleeding from the gastrointestinal tract represents relatively common diagnostic and therapeutic challenge in clinical work of gastroenterologists and surgeons. Bleeding from the lower GI (LGIB) is mostly caused by pathologic conditions of the colon, although the source of bleeding cannot always be exactly localized, thus rendering optimal and prompt therapy difficult. During two year period, at IlI department of the First Surgical Clinic in Belgrade, we performed 424 colonoscopies for LGIB. According to our results the exact diagnosis was established in about 76% (324 patients) showing a great similarity with the results of other published studies (varying between 74% and 89%). The most common causes of bleeding were diverticulosis (37.11%), polyposis (10.3%) and colorectal cancer (46.14%). Besides that we have mentioned some specific facts involving the diagnosis and treatment of LGIB with an accent on some rare conditions, like angiodysplasia. Review of the diagnostic procedures and treatment modalities of the LGIB is useful for everyone who meets with this type of pathology in clinical practice. The diagnostic approach and the surgical treatment of these patients may represent a great problem, since the planning of the operative procedure can be very difficult and with uncertain result. Based on the literary data and our experience we have tried to set the algorithm of the diagnostics and treatment of the LGIB. PMID- 18510059 TI - [Myocardial revascularization without extracorporeal circulation]. AB - Contemporary treatment of coronary disease includes: drug treatment, percutaneous coronary angioplasty (PCI), with or without stent implantation and surgical myocardial revascularization. For more than 30 years, conventional coronary bypass (on-pump CABG), using cardiopulmonary bypass (CPB), represented the standard regarding myocardial revascularization, particularly in patients suffering from three vessel disease or left main coronary artery stenosis. Recent development of invasive cardiology and increased interest in coronary surgery on the beating heart (OPCAB), challenging traditional on-pump CABG procedure, as optimal strategy for the treatment of coronary artery disease. In order to improve clinical outcome, OPCAB seems to be a good choice in patients with co morbidities critical for use of CPB. Results of OPCAB revascularization in general patient population are considerably different and require further evaluation. This review article shows the development of OPCAB and elaborates potential advantages and weaknesses of this method of revascularization, from both, theoretical and clinical point of view, compared to standard surgical myocardial revascularization. PMID- 18510060 TI - [Carotid artery plaque in patients with disorders of glucose regulation]. AB - PURPOSE: The aim of this study was measurement of artery intima media thickness (IMT) and plaques as an early indicator of atherosclerosis in diabetics comparing with other risk factors of carotid artery. METHODS AND MATERIALS: 110 pts: 50 with Diabetes Mellitus, type 1 (25) and type 2 (25), 20 pts with glucose intolerance, 20 pts with type 2 de novo and 20 pts obese without diabetes. Ultrasound examination (using 7.5 MHz sound on Toshiba SSA-270A) end measurement of intima-media ticknes (IMT) were performed on Carotis communis (CCA), bifurcation and distal from bifurcation to a.carotis intern (ACI), expressed in mm. Plaques were correlated with other common factors age, BP, lipid parameters (Chol, HDL, LDL, Triglycerides), smoks, alcoholism and obese (BMI). The authors used 2 test and Spearman's correlation. RESULTS: The lowest percent of plaques was found in group with type DM 1. The highest percent of plaques was found in type DM 2. Statistically there is highly significant difference between plaques founded on type 2 DM and types 2 DM de novo and on other types. CONCLUSION: DM is not an independent risk factor for developing of macroangiopathic changes an arterial walls, but their appirience are more presenting in diabetic patients. The highest number of plaques are presenting DM type 2 (29.6%), and after type 2 de novo (26.8%), the next highest position of plaques were in patients with obese but without DM and intolerantio glucosae (IFG+IGT) (17.1%) and type 1 DM (9.8%). Risk factors were presented in following percentage: Obese 80.5% pts; hyperlipidema 53.7% pts; HTA 51.3%; smoking 51.2% pts and alchocholism 2.4% pts. According to these results, all risk factors were included in patophysiology of plack forming except alcoholism. Influences of these risk faktors are very importance and their synergic action lids to their rapid appirience and clinical manifestations. DM has specific position in patophisiology of atherosclerosis. PMID- 18510061 TI - Gastroesophageal reflux in infants and children. AB - Gastroesophageal reflux disease (GERD) is the most common esophageal disorder and the most frequent reason why infants are referred to the pediatric gastroenterologist, affecting as much as 30% of the pediatric population. Presenting features of GERD in infants and children are quite variable and follow patterns of gastrointestinal and extra-esophageal manifestations that vary between individual patients and may change according to age. Patients may be minimally symptomatic, or may exhibit severe esophagitis, bleeding, nutritional failure, or severe respiratory problems. GERD is also complex for the diagnostic techniques required to assess its repercussions or explain its origin. Although different abnormalities in motility variables, such as lower eso-phageal sphincter (LES) function, esophageal peristalsis and gastric motor activity can contribute to the development of GERD, the degree of esophageal acid exposure represents the key factor in its pathogenesis. Esophageal pH monitoring, based on both the detection of acid reflux episodes and the measurement of their frequency and duration, has been regarded as the most sensitive and specific diagnostic tool for diagnosing reflux disease. The aim of this paper is to give a concise review for the clinicians encountering this specific disease in infants and children. PMID- 18510062 TI - Evaluation of the Alvarado score in the diagnosis of acute appendicitis. AB - This study has been performed in the Emergency center, Clinical centre of Serbia, during the period 01.03.2007-01.09.2007. We performed this study on 57 patients with diagnosis suspected for acute appendicitis (ages 16-70). Parameters that make the Alvarado score are the following: migration of pain, anorexia, nausea or vomiting, right lower abdominal quadrant tenderness, rebound tenderness in right iliac fossa, elevated temperature, leukocytosis, shift to the left of neutrophils. The aim of the work is to evaluate the Alvarado scoring system in diagnosis of the acute appendicitis. With all the patients Alvarado score has been determinate preoperatively, and diagnosis was confirmed by intraoperative finding and histopatological examination of the removed appendix. All the patients with score 7 or more were surgically managed. Specificity (positive predictive value) was 92.59 % in males and 76.67 % in females. The negative appendectomy rate was 7.41 % with the males and 23.33 % with the females. The values of the Alvarado score are significantly higher in the patients with acute appendicitis, compared with the patients of the other diseases. With the application of the Alvarado scoring system we can decrease postoperative morbidity and mortality. PMID- 18510063 TI - [Epidermoid splenic cysts in children and adolescents]. AB - Epidermoid cysts of the spleen are a rare lesion comprising less than 10% benign non-parasitic splenic cysts. Two boys and three girls, aged 13 to 24 years (mean 18.0 years) were diagnosed over a 4-year period. Presenting symptoms were dull, acute left hypochondrium pain and diffuse abdominal pain. Hemogram and routine analyses, as well as radiography were performed for the diagnosis. Ultrasound and CT confirmed the cystic nature of the lesion. Definitive diagnosis is made by pathological findings. Was performed splenectomy on one patient, and was performed a partial splenectomy on the other patients, in order to eliminate the symptoms produced by the cyst and prevent potential complications (postspleenectomiam sepsis). Patients were examined postoperatively. They were asymptomatic and with a normal spleen remnant detected by ultrasound and CT. Routine hematological data, blood clotting factors, and immunoglobulins were normal. PMID- 18510064 TI - [The incidence rate of thyroid microcarcinoma during surgery benign disease]. AB - Thyroid microcarcinoma are well-differentiated tumors less than 1 cm in diameter. A retrospective analysis was performed on patients operated of benign thyroid disease at the Center for endocrine surgery, Institute of endocrinology, Clical Center of Serbia in Belgrade, from January 1st to December 31st 2004, in order to establish the incidence of microcarcinoma. Indications for surgery were euthyroid multinodular goiter in 201 patients, thyroiditis in 31, thyroid adenoma in 178, Graves disease in 89 and Plummers disease in 79 patients. The results of this study, demonstrate that in 13.4% of the patients operated for goiter, 6.4% operated for thyroiditis, 5.6% for thyroid adenomas, 9.0% for Graves disease and 7.0% of the patients operated for Plumers disease, the presence of a microcarcinoma was noticed in the definitive histopathologic examination. The results obtained are in line with the current knowledge of high incidence of thyroid microcarcinoma. PMID- 18510065 TI - [Indications and results of resection surgical approach in localized type of bronchiectasis]. AB - Although the prevalence of bronchiectasis decreased significantly in developed countries, in less developed and in developing countries, it still represents a significant cause of morbidity and mortality. The localised form of bronchiectasis is the indication for surgical treatment if recurrent respiratory infections make normal life and professional activity impossible. Less frequently, the operation is necessary independently on the symptoms duration, if massive hemoptysis are life threatening for the patient. Compared with the period 10-15 years ago, the diagnostics of bronchiectasis changed in terms that bronchography has been replaced by high resolution CT scan. Owing to angiographic studies performed on sufficient number of patients, the patophysiology of bronchiectasis is furtherly highlited, but without significant changes in the process of patient selection. In the text, particular accent was given to situations that usually represent practical problems: billateral bronchiectasis, hemoptysis, bronchiectasis after pleural empyema, abscending bronchiectasis and bronchiectasis in children. The outcome of the surgical treatment is good in 90% patients, with operative mortality thatis comparable to that after lung resections for other indications. PMID- 18510066 TI - [Anatomical and functional outcome in nonoperative management of patients with fractures of the distal radial bone]. AB - Tracking of anatomical and functional results obtained after non-surgical treatment of the distal radius included 115 patients in total, of whom 51 cases were available to be analysed and processed within this work. Tracking of the results was based upon Anatomical Score System and Gartland-Werley Score System. The final numerical results obtained through Anatomical Score were the following:excellent in 12 cases (23.52%), good in 30 cases (58.82%), satisfactory in 8 cases (15.68%), dissatisfactory in 1 case (1.96%). According to the Gartland Werley Score System, the final results included: excellent in 19 cases (37.25%), good in 24 cases (47.00%), satisfactory in 7 cases (13.72%), dissatisfactory in 1 case (1.96%). This work also presents correlation between Residual Deformity score, Subjective Hardship score and Objective dysfunction, with the results obtained by Anatomical score and Gartland-Werley total score. The possibility of dissatisfactory results obtained by radiological screening followed by dissatisfactory outcome is described in literature; however, dissatisfactory screening results could be found in up to 31.7% of cases with satisfactory clinical findings. Our work records 15.7% of the latter cases. We might say that non-surgical treatment is not to be accepted as the only and exclusive approach to distal radius fracture. Thus, proper evaluation is mandatory. PMID- 18510067 TI - [Pathophysiological mechanism of the developing radicular cyst of the jaw]. AB - The radicular cysts are result of inflammatory process in the periapical tissues associated with necrotic and infected pulps. Humoral and cellular immune responses play a central role in the pathogenesis of these lesions. The most important role in the growth of these lesion have proinflammatore cytokine TNF alpha, IL-1 and IL-6. Cytokine can be secreted by macrophages, monocytes and other cells of the immune system and can participate in skeletal homeostasis including osteoclastic formation, and bone resorption in maxillofacial region. The aim of this study is to give a consise rewiew for mechanism of growth of maxilofacial radicular cysts, indicated of clinical aspect, as well as expalined role of cytokine in this pathophysiology process. PMID- 18510068 TI - [Hemodynamic effect of local infiltrative 2% lidocaine adrenaline anesthesia in general balanced anesthesia during middle ear surgery]. AB - Hemodynamic effect of different techniques and type of anesthesia are defined. The volume of reduced hemorrhage in surgical field is debatable, without any definite conclusion. The objective of the study is to investigate the effects of local infiltrative anesthesia with adrenaline during general balanced anesthesia and nitroglicerol on blood presure and hemorrhage reduction in middle ear operations. Prospective, randomized study included 58 adult patients planned for the otorhinolaryngological surgery. Studied group of patient (n 30) planned for middle ear operations. Befora incision surgeon was administered local infiltrative anesthesia using 2% lidocaine with adrenaline (1:200,000) The controls group (n 28), planned for other surgery had no local infiltrative anesthesia with adrenaline. Hemodinamic parameters were monitored before surgical incision and 30 minutes after that. Surgeon's verbal reply on hemorrhage reduction during surgery was recorded. Statistical analysis of parametric data was carried out by Mann-Whitney sum test. Blood pressure and heart rate was not different between the two groups. But, intraoperatively, the study showed lowering of sistolic and diastolic blood pressure. This mode of treatment and surgeon's verbal reply confirmed that local infiltrative anesthesia with adrenaline under balanced anesthesia had no effect on hemorrhage reductionin surgical field. PMID- 18510069 TI - [Management of cadaveric organ donors]. AB - Because the supply of cadaveric organ donors is limited and their ICU management is complex, a multidisciplinary, well-coordinated, and institutionally supported approach to management is essential to ensure the maintenance of the current supply and to increase the future supply of organs and tissues that are suitable for transplantation. The potential organ donor is at high risk for instability as a direct consequence of the loss of physiologic homeostatic mechanisms that are dependent on functioning of the central nervous system. The keys to successful ICU management of the potential organ donor include a team approach that is focused on the anticipation of complications, appropriate physiologic monitoring, aggressive life support, with frequent reassessment and titration of therapy. PMID- 18510070 TI - [Surgical or conservative treatment of penile fracture]. AB - INTRODUCTION: Penile fracture presents with rupture of corpora cavernosa. It is not rare but is remarkable. Treatment options are controversial. AIM: To establish the incidence, what are early and late complications in patients treated surgically or with conservative treatment options (prospective/retrospective and randomized analysis). To find out what therapy verified as superior. MATERIAL AND METHOD: The total of 32 patients with typical acute penile fracture were analyzed. The patients were between 20 and 56 years of age (Mean age 40+/-7.25 SD). A total of 11 patients that presented up to 48h after the rupture of tunica albuginea (34.37%) were operated, while 9 patients (28.13%) were conservatively treated. 12 patients (37.5%) reported to andrology consulting room due to penile fibrous alterations and/or erectile dysfunction 6 to 14 months after the anamnesticaly established, typical penile fracture ("primary latent penile fracture"). Both groups of patients were strictly clinically followed 3, 6, 9 and 12 months after the penile fracture. Eventual development of late complications (erectile dysfunction ED, penile curvature, induratio, caverno-urethral fistula and urethral stenosis) was also recorded. Etiology of ED was investigated: anamnesticaly, by hormone and laboratory analysis, using penodynamic test (PGE1), Duplex Doppler echosonography, and dynamic cavernosography. RESULTS: In the operated group of patients, (11 pts, 34.37%) preserved erectile capacity was recorded and no penile deformity or plaque lesions on the site of suture, after the patients were recommended to have 4 weeks of sexual abstinence. The rupture of tunica albuginea was always transversal in relation to penile axis i.e. in relation to axial force against which the penis acts during the intercourse. The rupture is always on the basis or mid penile portion . Out of 21 (65.63%) conservatively treated patients in 13 (61.9%) plaque lesions (longitudinal diameter 2 cm) was recorded, while sexual dysfunction was established in 13 patients (12 with ED and 1 patient with incapability for vaginal penetration). Evaluation of post therapeutical complications in relation to type of treatment, significantly higher frequency was recorded in the group of conservatively treated patients (p<0.01). Penile veno-oclusive dysfunction was registrated as significantly most frequent cause of ED (in 10-76.92% out of 13 patients). All patients with ED had palpable fibrous lesion similar to Peyrone's plaque, while the degree of penile deviation correlated to the size of plaque lesion. CONCLUSION: The late complications of penile fracture were significantly more frequently recorded in the group of conservative treated patients (p<0.01) which proves that this lesion presents as urgent urological entity. The penile rupture is always transversal, and most probable place of rupture is physiological penile curve since this is the site of weakened structural integrity. PMID- 18510071 TI - The first transendoscopic plastic to self-expandable metal stent replacement in Serbia and Montenegro using a diagnostic duodenoscope. AB - Studies have shown that SEMS remain patient longer with fewer associated complications compared with conventional plastic strents. Zilver stent, a nitinol zig-zag mash SEMS has a special advantage, having a thin introducer diameter of 7 Fr with a fully deployed span of 10mm. For a 48-year-old woman presented with obstructive jaundice and a diagnosis of unresectable pancreatic carcinoma with consequent stenosis of common bile duct (CBD), infiltration of local blood vessels and life expectancy longer than six months, it was decided that an endoscopic palliative drainage procedure should be performed. The technique of transendoscopic plastic to metal stent exchange is described, using a diagnostic duodenoscope. The patient lived 7 months after implementation of Zilver stent and died anicteric due to progression of a primary disease. The transendoscopic plastic to metal stent exchange is feasible palliative method which requires a basic endoscopic equipment and experienced staff and therefore is applicable in developing countries as well. PMID- 18510072 TI - [A laboratory multipositional skull base holder]. AB - Authors present an original multipositional holder for skull base cadaveric specimens, either formalin or fresh. It is constructed to be fixated for standard neurosurgical operating table, devoted for laboratory micro neurosurgical dissection and suitable for simulation operation on patient. The holder is made of solid stainless steel, practical and easy for cleaning. The device permits good tree point's fixation, extensive angular mobility of the primary and secondary joints, that the skull base specimen can be placed and supported in any desired position. It fulfills demands for fixation from external skull base surface with thin needles ensuring full surgical dissection field without obstacles or invisible angles. PMID- 18510073 TI - [The microbiology of peritonsillar abscesses]. AB - BACKGROUND: Peritonsillar abscess (quinsy) is a complication of acute bacterial tonsillitis. Its treatment remains controversial. One element of controversy is the choice of antibiotics after surgical drainage of the abscess. Results of many studies support the resistance of grown bacteria to many antibiotics and the potential importance of anaerobic species in development of peritonsillar abscesses. AIM: The purpose of the study was to investigate bacteriology of peritonsillar abscesses in the group of own patients in an attempt to establish optimal method of antibiotic treatment after drainage of the abscess. MATERIAL AND METHODS: Abscess material from 12 patients aged 20-43 years (mean: 31.5, s.d.: 6.8), 4 women and 8 men, with peritonsillar abscesses was obtained by aspiration and sent for aerobic and anaerobic cultures. All patients were subsequently treated with oral phenoxymethylpenicillin (4.5 million units per day) and metronidazole (1500 mg per day). RESULTS: A total 18 bacterial isolates (9 anaerobic and 9 aerobic and facultative) were recovered, accounting for 1.5 isolate per specimen. Anaerobic bacteria only were present in 3 patients, aerobic and facultatives in 3, and mixed aerobic and anaerobic flora in 6. Single bacterial isolates were recovered in 6 infections. The predominant bacterial isolates were Streptococcus and Bacteroides. Recovery in all examined subjects was complete. CONCLUSIONS: In the routine management of peritonsillar abscess, bacteriologic studies are unnecessary on initial presentation. It is, however, necessary to consider infection with anaerobes, hence we recommend penicillin and metronidazole as the antibiotic regimen of choice in the treatment of peritonsillar abscesses. PMID- 18510074 TI - [Application of Lugol solution in the gastroesophageal reflux disease]. AB - Endoscopy examination followed by Lugol solution staining is used in the diagnostics of early squamous cell carcinoma of the esophagus. The aim of the study was to describe usage of this method for the assessment of effectiveness in the gastroesophageal reflux disease. The method uses reaction between glycogen, present in epithelium, and iodine in Lugol solution. The study was conducted in 98 patients. Endoscopic assessment was made before application of Lugol solution, subsequently the gastroesophageal borderline and staining of mucosa membrane after application of Lugol solution was assessed. Biopsies were taken from the stained and unstained areas. The performed study showed that sensitivity of the method is 84%, specificity 79% and effectiveness 85%. The ratio of stained and unstained areas is statistically typical p=0.045. CONCLUSIONS: Endoscopic examination followed by Lugol solution staining are sensitive diagnostic methods in the gastroesophageal reflux disease. It is a simple and quick method which should be widely applied. PMID- 18510075 TI - [Dynamics of bone biochemical markers in nephrotic children treated with prednisone and metabolites of vitamin D]. AB - Children with idiopathic nephrotic syndrome (NS) are at risk for a disorder of bone metabolism. The role of metabolites of vitamin D supplements in prevention of this disorder has not been yet clarified. The aim of the study was to evaluate changes in selected bone biochemical markers in children with NS treated with different doses of prednisone and metabolites of vitamin D. METHODS: 40 patients with NS (mean age 9.2 +/- 3.6 years) treated with prednisone were evaluated in three groups depending on supplementation with metabolites of vitamin D: I--18 patients with alphacalcidol (0.05 mg/kg/week); II--10 patients with vitamin D (800 IU/day); III--12 patients without metabolites of vitamin D. In all patients, calciuria (Cau), phosphaturia (Pu), serum calcium (Cas), phosphate (Ps), parathyroid hormone (PTH), alkaline phosphatase (AP), carboxyterminal propeptide of type I procollagen (PICP), carboxyteminal pyridinoline cross-linked telopeptide of type I collagen (ICTP) and PICP/ICTP ratio were checked three times: T0--before prednisone treatment; T1--after 2 weeks of therapy with prednisone 2 mg/kg/24h; T2--after 2 weeks of treatment with prednisone 2 mg/kg/48h. RESULTS: Mean Cau and Pu values found at T1 were significantly higher and mean values of Ps, AP, PICP, ICTP concentration and PICP/ICTP ratio were significantly lower in 3 groups of pts in comparison to TO. A significant increase of Cas concentration was found in group I and control group, but there were no significant differences among values of Cas in 3 groups. In T2 period PICP and PICP/ICTP ratio increased (T2 vs T1) in all groups, however mean values of AP did not change significantly. Moreover, there was an increase of Cas and decrease of PTH in group I. In group I and II, levels of phosphaturia showed a significant difference among 3 studied groups. CONCLUSIONS: 1. The method of corticosteroid administration influences dynamics of selected bone biochemical markers in nephrotic children: short-term treatment with high daily doses is associated with increased calciuria and phosphaturia, decreased activity of alkaline phosphatase and suppression of serum markers of type I collagen's turnover (PICP, ICTP, PICP/ICTP ratio); decrease of corticosteroid dose leads to increase of these markers. 2. Decreased PICP/ICTP ratio during daily corticosteroid treatment may indicate stronger inhibition of bone formation than bone resorption. 3. Dynamic changes in studied bone biochemical markers in nephrotic children treated with prednisone do not depend on supplementation with vitamin D or alphacalcidol. PMID- 18510076 TI - [Causes of patients' dissatisfaction with family doctor care in the light of questionnaire surveys--analysis of responses to open-ended questions]. AB - The current study is based on questionnaire surveys investigating the quality of the family doctor care from the patient's perspective, conducted in Bialystok. The aim of this study was to identify the causes of patients' dissatisfaction with family doctor care. The object of analysis was written responses to two open ended questions included in the questionnaires. Content analysis was used. All the written responses were stored in the computer database and then grouped into categories. Of 361 respondents who answered the first open-ended question--128 (35.5%) gave negative comments, while among 166 respondents who replied to the second question--110 were dissatisfied (66.3%). Overall, 340 negative statements on the family doctor care were recorded. Three main categories of patient dissatisfaction were identified: 1. deficiencies in the primary health care system (81 statements); 2. deficiencies in organization and quality of services (80 statements); 3. deficiencies in care providers' attitudes, skills and work (179 statements). The analysis of responses has revealed situations in which patients are not satisfied with care. This information can be useful for improving the quality of the family doctor care. PMID- 18510077 TI - [The influence of ultraviolet radiation on HLA-DR+ and CD1a+ cells' number in systemic lupus erythematosus patients]. AB - Systemic lupus erythematosus (SLE) is a chronic, autoimmune disease with complex etiology in which genetic, hormonal and environmental factors including ultraviolet radiation play an important role. Immune and inflammatory processes participate in skin lesions development and internal organ damage. Till now most of investigations estimated the role of inflammatory and immune response in skin lesions, but only scarce data evaluate these parameters in normal appearing skin in SLE patients. The aim of our study was to determine the influence of high dose UVB on the number of HLA-DR+ and CD1a+ cells in normal appearing skin of SLE patients. The study included 20 SLE patients and 14 healthy controls. The number of HLA-DR+ and CD1a+ cells were estimated by immunohistochemical method in two skin biopsies taken from non-irradiated and from irradiated sites, 24 h after exposure to 3 MED. All skin sections were stained with antibodies directed against HLA-DR antigen and CD1a molecule. The number of HLA-DR+ was statistically significantly higher in SLE patients comparing to healthy individuals, and UVB irradiation enhanced the difference. Similarly, the mean number of CD1a+ epidermal cells was significantly higher than in controls. In healthy subjects ultraviolet radiation caused migration of these cells into dermis, thus their number was lower in the epidermis. In contrast, in SLE patients under UVB CD1a+ cell count was significantly higher. The increased number of HLA-DR+ and epidermal CD1a+ cells in SLE may suggest that enhanced antigen presentation, leading to excessive cellular immune response and inflammatory reaction development, may result in induction or photoinduction of LE specific skin lesion. PMID- 18510078 TI - [Anxiety, depression and social support in patients with psoriasis]. AB - Dermatological patients present significantly more psychological disturbances of various types than the general population. The aim of the present study was to analyze anxiety and depression in relation to social support in psoriasis patients. The study was performed on a group of 32 psoriasis vulgaris in-patients of the Department of Dermatology, Medical University of Lodz. The following questionnaires were employed: The Hospital Anxiety and Depression Scale (HADS) and Social Support Questionnaire (SOZU--Soziale Unterstutzung). Psoriasis severity was evaluated by Psoriasis Area and Severity Index (PASI). Elderly psoriatic patients presented significantly higher levels of both anxiety and depression. Moreover, lower depression levels were observed in patients with positive psoriatic family history. Furthermore, there was a positive correlation between anxiety level and cutaneous lesions severity, pruritus severity and longer disease duration. Additionally, there was a negative correlation between both anxiety and depression levels and social support. In conclusion, support groups should primarily be organized for elderly psoriatic patients with negative psoriatic family history. PMID- 18510079 TI - [Natural factors influencing sleep]. AB - Sleep is a universal phenomenon of human and animal lives, although the importance of sleep for homeo-stasis is still unknown. Sleep disturbances influence many behavioral and physiologic processes, leading to health complications including death. On the other hand, sleep improvement can beneficially influence the course of healing of many disorders and can be a prognostic of health recovery. The factors influencing sleep have different biological and chemical origins. They are classical hormones, hypothalamic releasing and inhibitory hormones, neuropeptides, peptides and others as cytokines, prostaglandins, oleamid, adenosine, nitric oxide. These factors regulate most physiologic processes and are likely elements integrating sleep with physiology and physiology with sleep in health and disorders. PMID- 18510080 TI - [Growth hormone deficiency in adults]. AB - Growth hormone (GH) deficiency in adults has only recently been recognised as a clinically important syndrome which is associated with increased morbidity and probably also with increased mortality. Its presence leads to many unfavourable changes in body composition (increased fat mass and decreased lean mass), lipid metabolism, bone mass and may worsen quality of life. In many patients the disease may be either asymptomatic or present with relatively moderate subjective symptoms. The signs and symptoms, if evident, however, are often too common to have a diagnostic value without a suggestive clinical context. Diagnosis of GH deficiency in adults is more difficult than in children. It should be always confirmed by the results of biochemical, especially provocative, tests, among which the insulin tolerance test is considered the test of choice. Long-term recombinant GH administration in GH-deficient adults improves body composition, muscle strength, quality of life, bone mass and density, and lipoprotein pattern. In this review article we discuss the causes, clinical picture, diagnosis and treatment of GH in adults and provide the reader with some practical guidance concerning dealing with a patient suffering from this disorder. PMID- 18510081 TI - [Evaluation of the surgical risk in lung parenchyma resection]. AB - Due to the high incidence of lung cancer and widely performed lung parenchyma resection, evaluation of respiratory function is a crucial aspect of patient's comfort and life quality. A method of choice is a surgical lung cancer treatment. Ablation of the lung tumor is related with loss of the active respiratory surface; thus evaluation of respiratory reserves and postoperative respiratory function is so an important aspect of patient qualification for a surgical treatment. Improvement of anaesthetic care along with surgical technique enables patients, with the higher surgical risk, to be qualified for surgery. PMID- 18510082 TI - [Role of the pathomorphologist in diagnostic process concerning problems of ortothopic liver transplantation in adults]. AB - In this paper are presented main tasks of the pathologist dealing with liver diagnosis concerning problems of its transplantation in adults. They include: pretransplant diagnosis of recipient's liver, examination of explanted liver, evaluation of donor liver (pretransplant biopsies and performed during the operation), posttransplant assessment of the graft. On the basis of our own experience, the most frequent problems in patologist's work and basic information concerning the diagnostic process in liver transplantation with posibillities and limits of the histopathological method are described. Because of the popularity of transplantation as the method of treatment of advanced liver diseases, the group of transplanted patients continously enlarges and may lead to visit to doctors of different specialties. That is why presentation of this issue seems to be very valuable. PMID- 18510083 TI - [Granular cell tumor of the larynx--literature review and case report]. AB - Granular cell tumor (Abrikosov's tumor) of the larynx is a very uncommon lesion. Up to date, only about 200 cases have been reported. The exact origin of granular cell tumor is still unclear, however most authors believe it to be a neoplastic process derived from neuroectoderm. Immunohistochemical positive staining for S 100 protein is found in all cases. Complete resection of the lesion with an attempt to leave remaining laryngeal structures intact, generally results in total recovery. The majority of reported tumors were removed by microlaryngoscopy. CASE REPORT: 63-year-old female presented 2-month-long history of increasing dysphonia. She had smoked 20 cigarettes a day for over 30 years. Indirect laryngoscopy disclosed oedematous mucosa in both vocal folds. No tumor was observed. Applying Kleinsasser procedure, a tumor 3 mm in diameter covered with normal mucosa was found growing in the posterior wall of the bottom of the right ventricle. The tumor as well as hypertrophic mucosa of the vocal folds were removed. Histopathological examination of the operative specimen revealed a granular cell tumor and chronic laryngitis. Immunohistochemically tumor cells were positive for S-100. The patient has been asymptomatic for 6 months. CONCLUSION: Surgical removal of the laryngeal granular cell tumor with indications and extent of the resection like in benign tumors, was therapeutically efficient in the presented case. PMID- 18510084 TI - [Meningiomas of the anterior cranial fossa: clinical and radiological presentation--report of 2 cases]. AB - We reported two cases of the anterior cranial fossa meningiomas: cerebral falx meningioma and recurrence of olfactory groove meningioma. Since the tumors grow very slowly, they remain clinically undetectable during the early stages and can reach a very large size. Clinical manifestations are caused by pression of meningiomas on adjacent structures. The authors present possibilities of radiological examinations of intracranial tumors and treatment management. In the described cases meningiomas were diagnosed in CT examination with the use of angio option and reconstruction RT3D and MPR. PMID- 18510085 TI - [Przeglad Lekarski (Medial Review). The difficult years]. AB - The progress of medical science with great difficulties in the Polish territories in the second half of 19th century is presented in short. The achievements of such Cracovian physicians as: Jozef Dietl, Jozef Oettinger, Antoni Bryk, Jozef Majer, Fryderyk K. Skobel, Michal Zieleniewski is the creation of the Cracovian periodical "Przeglad Lekarski" ("Medical Review") is briefly pointed out. The critical observations of such Polish physicians as Jozef C. Starkel, Jozef Oettinger, Boleslaw Placer, Adolf S. Friedberg in the 1860s of the 19th century were very helpful in the development of this medical magazine. PMID- 18510086 TI - [Dear friends and colleagues]. PMID- 18510087 TI - [Nerve sparing radical hysterectomy]. AB - The aim of the study was to review the literature concerning nerve sparing radical hysterectomy (NSRH). The data about anatomical and physiological background of this operation, its history, technique and perspectives has been presented. In conclusion it has been estimated that the technique is new and its oncological efficiency is not yet fully established; it requires special anatomical knowledge and new instruments. PMID- 18510088 TI - [Selected sexuality features among married women--research results]. AB - OBJECTIVES: Analysis of features describing sexuality and sexual behavior of married women, deeming their sexual life 'successful' or 'unsuccessful'. MATERIAL AND METHODS: 217 women have been interviewed with the help of our original questionnaire about sexual life. All women in question had been married at the time of the interview. The results have been submitted for statistical analysis. RESULTS: Among 217 married women that had been interviewed, 190 (87.6%) described their sexual life as 'successful', while 25 (11.5%) as 'unsuccessful'. Two women were not sexually active and thus have been excluded from the study. CONCLUSIONS: 1. Women in failed marriages discuss the causes of their problems with their husbands less frequently, and more often have unrevealed sexual expectations and dreams, in comparison to the group of women in successful marriages. 2. Women who deemed their sexual life 'unsuccessful' have problems with accepting nudity, both theirs and their husbands. 3. Women describing their sexual life as 'unsuccessful' do not perceive sex as pleasure. 4. The belief that one has a 'successful' or 'unsuccessful' sexual life does not have influence on the attempts of undertaking sexual contacts, with or without the consent of a woman. 5. Almost a half of married women giving consent for intercourse against their will, both in 'happy' or 'failed' marriages, are of the opinion that a husband has the right to rape his wife. 6. 'Successful' sexual life in a marriage is not a guarantee of woman's faithfulness. 7. The type of contraceptive used does not have an influence on perceiving one's sexual life as 'successful' or 'unsuccessful'. PMID- 18510089 TI - [CA 125 regression after two cycles of neoadjuvant chemotherapy as prognostic factor in patients with advanced ovarian cancer and primary peritoneal serous cancer who underwent interval surgical cytoreduction]. AB - OBJECTIVES: The aim of our study was to assess the prognostic role of CA 125 regression during neoadjuvant chemotherapy (NAC) in patients with ovarian cancer (OC) or primary peritoneal serous carcinoma (PPSC) that underwent interval debulking surgery (IOC). MATERIAL AND METHODS: Thirty one patients with advanced OC or PPSC (FIGO stage IIIC and IV) who underwent initial exploratory surgery, followed by NAC containing platinum analogs, have been analyzed, retrospectively. We have used a regression coefficient (RCA 125), which was calculated as following: log10 (CA 125 level measured after two cycles of NAC/baseline CA 125) for statistical analysis. The median value of RCA 125 reached -0.788 and has been used to dichotomize. Optimal IOC has been performed in 67.74% (21/31) patients, suboptimal in 25.81% (8/31) patients and 6.45% (2/31) of patients did not undergo IOC due to the progression of the disease. RESULTS: We have noted significant correspondence between time to progression and RCA 125 in univariate analysis, which we have also confirmed in multivariate analysis (HR 0.27; 95% CI, 0.15 0.96; p = 0.0178). Similarly, we have observed significant relationship between overall survival, RCA 125 and extension IOC in univariate analysis. Multivariate analysis confirmed that RCA 125 was independent prognostic factor, HR-0.18 (95% CI, 0.07-0.56; p = 0.004). In case of patients with high RCA 125, a greater rate of optimal debulking cytoreduction (p = 0.0278, U = 50.0) has been observed. CONCLUSIONS: RCA 125 after two courses of NAC appears to be an important prognostic factor in patients with OC or PPSC, who underwent IOC High RCA 125 during NAC seems to be a good predictive factor in order to achieve optimal IOC. PMID- 18510090 TI - [Quality of life versus semen parameters]. AB - INTRODUCTION: Psychological factors are believed to play an important role in infertility and its treatment Psychological problems may be considered to be risk factors and the cause of reduced infertility, as well as contribute to an unsuccessful outcome of infertility treatment. OBJECTIVES: The aim of the study was to investigate the correlation between semen parameters and quality of life in male patients with fertility problems. MATERIAL AND METHODS: 70 patients with infertility treated in Andrology Clinic were divided into two groups according to semen parameters: bad and good quality of semen. The quality of life has been with the help of Campbell questionnaires. RESULTS: Significantly lower questionnaire score in bad quality of semen group has been observed, in comparison with normozoospermic men. High correlation between concentration, motility and morphology in comparison with the quality of life has been observed in the studied group of patients. CONCLUSION: The quality of life is directly associated with semen parameters such as concentration, motility and morphology of sperm. PMID- 18510091 TI - [Assessment of the usefulness of ultrasound screening in fetal ovarian cysts]. AB - OBJECTIVES: The aim of the study was to assess the usefulness of ultrasound in management and prognosis in fetal ovarian cysts. MATERIAL AND METHODS: The study included 38 fetuses with cyst in abdominal cavity, who, between 1995 and 2006, underwent an ultrasound examination in our unit at the Polish Mother's Memorial Hospital in Lodz, The Department for Diagnosis and Prevention of Birth Defects. RESULTS: In all 38 fetuses with cyst in abdominal cavity we have diagnosed 27 (74%) cases of ovarian cyst. In 14 (74%) fetuses cysts regressed spontaneously, including all cysts < or = 40mm (n = 7). In 3 cases with cysts > 40mm needle aspiration has been successfully performed, without any further complications. Surgical neonatal treatment has been performed in 5 cases in prenatal cysts > 40mm without prenatal aspiration. In 3 cases cysts > 40mm regressed spontaneously. Ovarian cysts in 22 (87%) cases were an isolated malformation; in 5 (19%) cases other malformations were present CONCLUSIONS: 1. Fetal ovarian cysts < or = 40mm required only ultrasound assessment and, in majority of cases, revealed the tendency to spontaneous regression. 2. Cysts > 40mm in maximal diameter have signaled complications more often and required surgical procedure after birth. 3. In utero, aspiration of fetal ovarian cyst > 40mm may lead to cyst regression, making the surgery after birth unnecessary. PMID- 18510092 TI - [HPV infection in etiology of uterine cervix cancer]. AB - Uterine cervical cancer may be the direct result of persistent HPV infection. The DNA analysis shows over 200 HPV types. The natural history of uterine cervix cancer is a constantly progressing process, from low pathology changes, through medium to major pathology changes, including microinvasion and invasive carcinoma. The population studies show that over 20% of women at the moment of examination is infected with HPV. The study of cervical cancer tissue revealed the HPV in the DNA, whereas the serological study revealed the presence of specific antibodies to HPV capsid antigens in most women (in USA the number amounts up to 70%). The serological conversion after HPV infection is not a common process. Therefore, the naturally infected women, still do not have enough antibodies and they are at the risk of infection, just as seronegative women. The vaccination of women between the ages of 15 and 25, causes the increase of the number of antibodies, several times higher than in the case of naturally acquired ones. The positive results of HPV examination is not a contraindication to vaccination. However, in case of vaccinated women, the negative test for 14 oncogenic HPV types has an overwhelmingly negative predictive value. Independent of the presence or absence of HPV infection, it is essential to continue cytological. The vaccination does not protect patients from all oncogenic HPV types. Cytological screening enables doctors to recognize cervical pathology at the early stage and introduce a proper treatment. PMID- 18510093 TI - [Endometriosis in pediatric and adolescent gynecology]. AB - Endometriosis is the most common cause of chronic pelvic pain in adolescent girls (50-70%), unresponsive to treatment of oral contraceptives and non-steroidal anti inflammatory drugs. The most common symptoms of the disease are: acquired or progressive dysmenorrhea, acyclic and cyclic pain, dyspareunia (in sexually active girls), urological symptoms and gastrointestinal complaints. When evaluating an adolescent with suspected endometriosis, a gynecological examination (rectal or vaginal examination) and imaging studies (ultrasonography, magnetic resonance) should be performed. Moreover, in diagnostic process laparoscopy should be carried out in all girls and teenagers with chronic pelvic pain unresponsive to medical treatment. Initial therapy of endometriosis in adolescent girls involves: surgical methods (laparoscopy/laparotomy), hormonal pharmacotherapy (combined contraceptives, progestin-only protocols), GnRH agonists (adolescents over 16 years of age), non-steroidal anti-inflammatory drugs, alternative pain therapies and psychotherapy. Early diagnosis and treatment during adolescence may decrease disease progression and prevent subsequent infertility. PMID- 18510094 TI - [The influence of anti-Mullerian hormone on folliculogenesis]. AB - The main biological role of the anti-Mullerian hormone (AMH) is to induce the involution of the Muller ducts in embryos during differentiation of masculine gender. In case of women, AMH is produced in granular cells of primary, preantral and antral follicles. The expression of AMH initiates at the moment of the follicle recruitment and it lasts until the stage of an antral follicle. The level of this hormone decreases with age and in postmenopausal period is undetectable in blood. Therefore, AMH could be a useful marker of ovarian reserve. Multiple investigations have revealed higher AMH levels in the blood of PCOS patients. It is believed to be the consequence of the increased amount of small antral follicles. AMH is considered to have an essential role in folliculogenesis. It inhibits the process of recruitment of primordial follicles and modifies the growth of preantral and antral follicles by diminishing the sensitivity of follicles for FSH stimulation. The paper is a review of the present knowledge of the structure and activity of AMH. AR gene and protein. Participation of AMH in folliculogenesis and changes of AMH levels depending on structure and age of the ovary have also been discussed. Recent findings concerning the possibility of using AMH to assess ovarian reserve and efficiency of the stimulation of ovulation in infertile women have been presented. It is believed that increased knowledge concerning AMH might improve the diagnosis and treatment of infertility caused by lack of ovulation. PMID- 18510095 TI - [Abdominal Burkitt lymphoma mimicking the ovarian cancer. Case report and review of the literature]. AB - Primary Burkitt lymphoma is a lymphoblastic B-cell malignant tumor with very aggressive course. Its abdominal form involving internal genital organs is very rare. CASE: We report the case of 27-year-old woman treated for abdominal Burkitt lymphoma. The patient presented bilateral ovarian tumors with ascites, pain and elevated CA 125 over 900 IU/ml. During laparotomy an advanced neoplasmatic disease involving internal genital organs has been diagnosed. Bilateral salphingo oophorectomy and omentectomy have been performed. Additionally, the neoplasmatic tumor from ileo-coecal region has been ressected in order to prevent ileus. Pathologic examination has revealed an abdominal Burkitt lymphoma. After surgery, polychemotherapy has been administered (COP followed by CODOX-M+IVAC). The patient, 36 months after surgical treatment, remains under the control of our Department. No signs of recurrence have been detected so far. CONCLUSIONS: The presence of primary abdominal Burkitt lymphoma may include clinical and laboratory findings suggesting the presence of ovarian cancer. Chemotherapy appears to be an essential therapeutic management for all forms of Burkitt lymphoma. Clinically advanced Burkitt lymphoma may be successfully managed with chemotherapy. PMID- 18510096 TI - [Avoiding common mistakes in FHR monitoring. (Part II)]. PMID- 18510097 TI - PKC isotype functions in T lymphocytes. AB - The main function of mature T cells is to recognize and respond to foreign antigens by a complex activation process involving differentiation of the resting cell to a proliferating lymphoblast actively secreting immunoregulatory lymphokines or displaying targeted cytotoxicity, ultimately leading to recruitment of other cell types and initiation of an effective immune response. In order to understand the physiology and pathophysiology of T lymphocytes, it is necessary to decode the biochemical processes that integrate signals from antigen, cytokine, integrin and death receptors. The principal upon which our work is based is to explore and identify gene products of distinct members of the AGC family of protein serine/threonine kinases as key players mediating cell growth regulation. Given the established important role of PKC theta as regulator of T cell fate and knowing that several other PKC isotypes are also expressed in T cells at a high level, we now summarize the physiological and non-redundant functions of PKC alpha, beta, delta, epsilon, zeta and theta isotypes in T cells. This review describes the current knowledge of the physiological and non redundant functions of the PKC gene products in T cells. PMID- 18510098 TI - Systems biology of T cell activation. AB - T lymphocytes are central players in the adaptive immune response to pathogens. Cytotoxic T cells are able to identify and eliminate virally infected cells, while helper T cells support B lymphocyte-dependent antibody production as well as produce the cytokines that will determine whether a cell- or antibody-mediated immune response is required. The activation of T cells by pathogens is a complex process requiring multiple tightly regulated signaling pathways. Defects within this network, however, can cause severe and chronic disorders such as autoimmunity. Therefore, improving our understanding of how T cells discriminate between antigens and how these signals are organized to yield distinct immune responses is of importance as this may lead to the identification of novel drug targets and better therapeutic strategies. PMID- 18510099 TI - Solving the IRAK-4 enigma: application of kinase-dead knock-in mice. AB - Interleukin-1 receptor-associated kinase (IRAK-4) is an essential component of the signal transduction complex downstream of the interleukin (IL)-1- and Toll like receptors. Though regarded as the first kinase in the signaling cascade, the role of IRAK-4 kinase activity versus its scaffold function has been controversial. In order to investigate the role of IRAK-4 kinase function in vivo, we generated "knock-in" mice where the wild-type IRAK-4 gene is replaced with a mutant gene encoding kinase-deficient IRAK-4 protein (IRAK-4 KD). IRAK-4 kinase is rendered inactive by mutating the conserved lysine residues in the ATP pocket essential for coordinating ATP. Analyses of embryonic fibroblasts and macrophages obtained from IRAK-4 KD mice demonstrated lack of cellular responsiveness to stimulation with IL-1beta or Toll-like receptor 4 (TLR4) and TLR7 agonists. IRAK-4 KD cells were severely impaired in NF-kappaB, JNK, and p38 activation in response to IL-1beta or TLR7 ligand. In addition, activation of JNK and p38 was affected in lipopolysaccharide (LPS)-stimulated IRAK-4 KD macrophages. As a consequence, IL-1 receptor/TLR4/TLR7-mediated production of cytokines and chemokines was largely absent in these cells. Additionally, microarray analysis identified IL-1beta response genes and revealed that the induction of IL-1beta-responsive mRNAs is largely ablated in IRAK-4 KD cells. In summary, our results suggest that IRAK-4 kinase activity plays a critical role in IL-1R-, TLR4-, and TLR7-mediated induction of inflammatory responses. PMID- 18510100 TI - Sensing, presenting, and regulating PAMPs. AB - Recognition of microbial infection and initiation of immune responses are controlled by multiple mechanisms. Toll-like receptors (TLRs) are key components of the innate immune system that detect microbial infection. TLR activation helps to eliminate the invading pathogens, coordinate systemic defenses, and initiate adaptive immune responses. Despite progress elucidating the TLR signaling aspects and the physiological relevance of TLRs in microbial infections, the molecular basis of microbial recognition by TLRs is still not fully understood. In this article we focus on the availability of microbial ligands to regulate presentation to TLRs and assist in our understanding of TLR-mediated microbial recognition. PMID- 18510101 TI - Migration, cell-cell interaction and adhesion in the immune system. AB - Migration is an essential function of immune cells. It is necessary to lead immune cell precursors from their site of generation to the places of maturation or function. Cells of the adaptive immune system also need to interact physically with each other or with specialized antigen presenting cells in lymphatic tissues in order to become activated. Thereby a complex series of controlled migration events, adhesive interactions and signalling responses is induced. Finally cells must be able to leave the activating tissues and re-enter the bloodstream from which they extravasate into inflamed tissue sites. Cells of the innate immune system can function directly without the need for previous activation. However, these cells have to adapt their function to a panoply of pathogens and environmental niches which can be invaded. The current review highlights the central aspects of cellular dynamics underlying adaptive and innate cellular immunity. Thereby a focus will be put on recent results obtained by microscopic observation of live cells in vitro or by intravital 2-photon microscopy in live animals. PMID- 18510102 TI - The role of diacylglycerol kinases in T cell anergy. AB - Engagement of the T cell antigen receptor (TCR) results in the activation of multiple biochemical second messenger cascades that must be integrated for the appropriate T cell response. Once the critical TCR-stimulated signaling pathway is initiated by activation of protein tyrosine kinases, a series of adapter proteins is recruited that brings tyrosine-phosphorylated phospholipase Cgamma1 into the vicinity of its substrate, phosphatidylinositol-4,5-bisphosphate, resulting in the formation of two second messengers, inositol-1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Previous work from multiple laboratories has shown that the balance between signals downstream of IP3 versus those downstream of DAG has profound effects on the fate of the stimulated T cells. In this report we summarize our recent data indicating that one key determinant of this balance of signals is the activity of members of the diacylglycerol kinase family, enzymes that convert DAG into phosphatidic acid. PMID- 18510103 TI - Fragment-based drug discovery using rational design. AB - Fragment-based drug discovery (FBDD) is established as an alternative approach to high-throughput screening for generating novel small molecule drug candidates. In FBDD, relatively small libraries of low molecular weight compounds (or fragments) are screened using sensitive biophysical techniques to detect their binding to the target protein. A lower absolute affinity of binding is expected from fragments, compared to much higher molecular weight hits detected by high throughput screening, due to their reduced size and complexity. Through the use of iterative cycles of medicinal chemistry, ideally guided by three-dimensional structural data, it is often then relatively straightforward to optimize these weak binding fragment hits into potent and selective lead compounds. As with most other lead discovery methods there are two key components of FBDD; the detection technology and the compound library. In this review I outline the two main approaches used for detecting the binding of low affinity fragments and also some of the key principles that are used to generate a fragment library. In addition, I describe an example of how FBDD has led to the generation of a drug candidate that is now being tested in clinical trials for the treatment of cancer. PMID- 18510104 TI - [Patient with red ears. Untreated this could be fatal]. PMID- 18510105 TI - [Hardly any strength in the left hand. Where is the suspected damage?]. PMID- 18510106 TI - [Simply rinsing out pollen. Does nasal irrigation help control hay fever? (interview by Dr. Judith Neumaier)]. PMID- 18510107 TI - [No system exit from the united front after all. Bavarian family physicians waver before the final break]. PMID- 18510108 TI - [Antihypertensive agents, heart valve operation, stem cells: can heart failure be prevented?]. PMID- 18510109 TI - [Fatal interaction. Ailing psyche--weak heart]. PMID- 18510110 TI - [Neurologic problems in the elderly. Not jumping to diagnostic conclusions!]. PMID- 18510111 TI - [Will there soon be a sub-type specific treatment? New approaches to multiple sclerosis therapy]. PMID- 18510112 TI - [For many years chronic lymphatic leukemia: suddenly the illness "explodes"]. PMID- 18510113 TI - [Sleepless in a 24-hour society. When inner and external rhythms collide]. PMID- 18510114 TI - [Shift worker syndrome]. PMID- 18510115 TI - [Jetlag--prevention and therapy]. PMID- 18510116 TI - [Treatment of hypertension in patients with renal dialysis]. PMID- 18510117 TI - [Atrial fibrillation]. PMID- 18510118 TI - [Modern treatment of erectile dysfunction]. PMID- 18510119 TI - [Treating distal ulcerative colitis intrarectally. Foam instead of enema, low volume rather than high volume]. PMID- 18510120 TI - [Infrequent but serious: autoimmune hepatitis and primary biliary cirrhosis. Early therapy improves long-term prognosis]. PMID- 18510121 TI - Health care reform in the 2008 presidential primaries. PMID- 18510122 TI - Patient-staff interactions and mental health in chronic dialysis patients. AB - Chronic dialysis imposes ongoing stress on patients and staff and engenders recurring contact and long-term relationships. Thus, chronic dialysis units are opportune settings in which to investigate the impact of patients' relationships with staff on patient well-being. The authors designed the present study to examine the degree to which perceptions of open communication between patients and staff affect patient mental health. A one-year, two-wave longitudinal survey assessed patient (N = 109) perceptions of the interpersonal environment and mental health. Assessments included sharing personal information (open disclosure), assisting one another (helping), staff respect for patients (respect), and hierarchical patient-staff relations (formal staff authority). Cross-sectional and longitudinal regression analyses examined how these characteristics of the interpersonal environment relate to depression and subjective well-being among patients. Multivariate analysis showed that open disclosure correlated independently with lower levels of depression at baseline (N = 109) and a predicted significant decrease in depression over time (N = 64). Other interpersonal characteristics did not correlate with depression or subjective well-being at baseline or longitudinally. The interpersonal climate in chronic dialysis units influences patient well-being. Contrary to traditional views, open disclosure in patients' relationships with staff is not detrimental and contributes to well-being. PMID- 18510123 TI - The experience of living kidney donors. AB - This article describes the experiences, feelings, and ideas of living kidney donors. Using a phenomenological, qualitative research approach, the authors interviewed 12 purposefully selected living kidney donors (eight men and four women), who were between four and 29 years since donation. Interviews were audiotaped, and transcribed verbatim, and the analysis of the data was both iterative and interpretive. Three key themes emerged. The first was how witnessing their loved ones' experience of illness and the threat of losing the recipient influenced the participants' decision to donate. The second focused on intrapersonal (philosophy of life) and interpersonal factors (comprehensive social support networks) that influenced the decision to be tested as a potential donor and the actual process of donation. The third was the impact of giving the gift of life, which was emotional and life changing. This article provides a rich description of the experiences of living kidney donors, revealing the multiple factors influencing the decision to donate, and provides insight on how social workers and other health care professionals need to identify and address the psychosocial needs of living kidney donors and their families from the process of decision making through after donation. PMID- 18510124 TI - The diverse faces of Latinos in the Midwest: planning for service delivery and building community. AB - Throughout the Midwest and the southern United States, new groups of Hispanic/Latino and other immigrants are settling in large numbers. In many regions, very little infrastructure exists in social service, health care, and educational entities that would allow professionals to deliver much-needed services. Little data exist on the sociodemographics of new immigrant communities in the South and the Midwest. Sociodemographic information provides social work professionals and community planners with valuable clues about the health and social services that may be necessary to promote a viable, livable community. In this article, the authors report on a survey of 535 Hispanic adults conducted in a midwestern city, surrounded by urban, suburban, and rural areas in three adjacent states. This information can be used to design programs and services to benefit new immigrants and to assist communities in predicting future needs. PMID- 18510125 TI - Cultural competence in a group intervention designed for Latino patients living with HIV/AIDS. AB - Although the trajectory of the HIV/AIDS epidemic has changed dramatically over the past 25 years, addressing the psychosocial needs of patients living with HIV/AIDS remains vital. Ensuring the effective delivery of services demands that interventions be rooted in cultural competence and aimed at vulnerable populations. This article describes a group intervention designed for Latino patients living with HIV/AIDS in NewYork City. The intervention effectively integrates culturally competent practice with traditional social work practice with groups' skills to provide an arena for participants to explore issues commonly faced by patients living with HIV/AIDS in a cultural context. Case examples are used to describe themes that emerged during the intervention, which illustrate cultural influences on issues such as adherence, social isolation, stigma, disclosure, safer sex practices, and patient-provider communication. Cultural factors inherent to Latino culture that are known to influence a patient's health experience and the development of effective interventions are also presented. The identification and explanation of these factors is intended to help social workers and other health care professionals better understand the roots of Latino patients' beliefs and understanding of their HIV/AIDS condition as well as highlight some of the obstacles they cause. PMID- 18510126 TI - Self-control, self-efficacy, role overload, and stress responses among siblings of children with cancer. AB - The study focuses on healthy children's responses to a sibling's cancer and its aftermath, with particular scrutiny directed toward these healthy siblings' stress factors, duress responses, and coping resources. The authors investigated role overload as these siblings' stress factor, anxiety and psychosomatic symptoms as their duress responses, and self-control (SC) and self-efficacy (SE) as their coping resources. Participants comprised 100 (53 boys and 47 girls) Israeli Jewish healthy siblings (ages 8 to 19 years) of a child with cancer. Outcomes revealed that the stress experienced by healthy siblings ofa child with cancer correlated significantly with those siblings' duress responses: Greater role overload was linked with higher levels of state anxiety and more psychosomatic symptoms. Likewise, these siblings' stress factor correlated significantly with one of their personal resources: Greater SC was linked with lower role overload. Furthermore, personal coping resources correlated significantly with healthy siblings' duress responses: Greater SC and SE were linked with lower levels of anxiety and fewer psychosomatic symptoms. In addition, hierarchical regression analyses showed that, among children older than age 12, greater SC was linked with milder anxiety. Limitations and implications are discussed. PMID- 18510127 TI - Perceptions of biopsychosocial services needs among older adults with severe mental illness: met and unmet needs. AB - This study sought to identify the psychiatric, physical, and social services needs experienced by older adults with severe mental illness (SMI) and to examine factors influencing their experience of need and service provision adequacy. Seventy-five older adults with SMI were recruited from a community mental health center to participate in the study. The typical client experienced a need for care in 10 areas, with the greatest needs occurring in the areas of psychological pain, physical illness, social contacts, looking after the home, and daily activities. The total number of unmet needs ranged from zero to 10, with the typical client having an average of 2.3 unmet needs (SD = 2.4). The highest proportions of unmet needs were in the areas of social contact, benefits, sight or hearing difficulties, and intimate relationships. Linear hierarchical regression analyses revealed that clients with lower income, greater impairments in independent daily living skills, and higher levels of depression experienced increased needs for care. Older clients who lived in private homes or apartments had higher levels of depression, and those who required assistance in the areas of intimate relationships and benefits experienced higher levels of unmet needs. Research and practice implications are discussed. PMID- 18510128 TI - Prisoner reentry. PMID- 18510129 TI - Nurse social work practitioner: a new professional for health care settings. PMID- 18510130 TI - "I can't stop pulling my hair!" Using numbing cream as an adjunct treatment for trichotillomania. PMID- 18510131 TI - Relationships between personality variables and bizarreness effects in free recall. AB - Relationships between 6 personality variables and each of 3 different measures of recall for bizarre and common sentences were examined. The personality variables investigated included measures of sensation seeking, novelty experiencing, desire for novelty, arousal-seeking tendency, social potency, and conservatism. Recall was measured in terms of sentences accessed, target words recovered per accessed sentence, and misplaced target words. The results indicated the typical pattern of bizarreness effects on recall and significant relationships between personality variables and these effects. Arousal seeking and conservatism were positively related to a bizarreness advantage in sentences accessed. Additionally, high social potency was related to the recovery of more details from common than bizarre sentences, and high desire for novelty was related to a greater bizarre misplacement effect. The results are discussed in terms of orienting and defensive responses to bizarreness. PMID- 18510132 TI - Strategy use in mental subtraction determines central executive load. AB - A dual task method was used to examine the relationship between strategy use and working memory load during subtraction problem solving. Undergraduates mentally solved subtraction problems alone and while performing secondary tasks that involved the central executive of working memory. Analyses revealed that a central executive task involving response selection and input monitoring (CRT-R task) interfered more with subtraction problem solving than a task that involved only input monitoring (SRT-R task). Additional analyses showed that the CRT-R task interfered more when participants used a nonretrieval (counting) strategy than a retrieval strategy. These findings suggest that the response selection subcomponent of the central executive is involved during both retrieval-based and non-retrieval-based simple subtraction problem solving but is involved more during the latter. PMID- 18510133 TI - Regularity of symmetry verticality guides perceptual judgments of objects. AB - Previous research indicated that most salient, real-world objects possess natural regularities that observers commonly assume in perceptual judgments of figural orientation and interpretation. Regularities include 3-dimensionality, bilateral symmetry, and the tendency for object tops to possess more salient information than bottoms. Thus, when observers interpret randomly shaped figures, they reliably impose volume, bilateral symmetry, and top and front orientation directions, even when figures are 2-dimensional and asymmetric. We confirmed generalizability for observers to assume these regularities with stimuli that vary in complexity, and we found evidence supporting another regularity, that of symmetry verticality (symmetry about a vertical axis). Findings support use of a family of perceptual heuristics corresponding to natural regularities that constrain stimulus indeterminacy and help guide judgment of object orientation and interpretation. PMID- 18510134 TI - On the spatial metric of short-SOA costs of exogenous cuing. AB - When the stimulus onset asynchrony (SOA) between the cue and the target is short, exogenous spatial cues usually produce a response time benefit. However, consistent with several recent studies, we have found that a short stimulus onset asynchrony is not sufficient. At least one more factor--the number of cue and target locations--also plays a role. Even more interesting, when 8 cue and target locations are used, the effect of an exogenous cue produces a cost on valid cue trials, and the spatial metric of this negative cuing effect depends on whether the cue remains visible at target onset. PMID- 18510135 TI - Exploring students' prospective memory inside and outside the lab. AB - This study compared activity-based intentions with event- and time-based intentions. Forty-five participants completed a laboratory session that tested activity-, event-, and time-based prospective memory. Participants were then asked to keep a record of their planned academic and work-related activities for 1 week. Results revealed a main effect of context such that participants were better at performing intentions in the naturalistic setting than in the laboratory. A strong trend of intention type showed that event-based tasks were more likely to be performed than activity- and time-based tasks. An interaction of context and intention type suggested that participants were significantly better at performing time-based tasks in the naturalistic setting than in the lab. Nearly half of the participants reported using simple memory aids such as alarm clocks, parent or friend reminders, or other external cues. Strikingly, very few reported using electronic devices to aid their memory for future intentions. PMID- 18510136 TI - Thinking styles and identity development among Chinese university students. AB - This study investigates the relationship between thinking styles and identity development among Chinese students. Students from Shanghai responded to the Thinking Styles Inventory-Revised, based on Sternberg's theory of mental self government, and to the Erwin Identity Scale-III, grounded in Chickering's theory of psychosocial development. Not only were thinking styles strongly associated with identity development, but also the former had predictive power for the latter. Along with findings in the existing literature, results of this study call for a collaborative effort between teaching faculty and student development educators in fostering holistic student development. PMID- 18510137 TI - Measuring cognitive distance in the network representations of texts. AB - Pairs of words are identified more quickly as coming from different sentences the further apart the words are in the network representation of their native text. How is the distance between the words measured? According to the network connection hypothesis, the distance between 2 words is the length of the network path between the propositions to which they belong, as measured by a process of spreading activation. According to the location code hypothesis, the distance between 2 words is the difference between codes that record the locations of their propositions in the network representation. The results of 3 experiments supported the network connection hypothesis. PMID- 18510139 TI - Surviving a split-reporting relationship. PMID- 18510138 TI - Remembering others: using life scripts to access positive but not negative information. AB - The current research extended to memories of others the life script theory of abstract, idealized mental representations of transitional experiences. Recent and earlier high school graduates rated positive and negative characteristics of popular, average, and unpopular girls from their schools. "Average" girls were rated as higher than average on possessing positive characteristics. Recent but not earlier graduates distinguished between popularity conditions on negative characteristics (negative information is not included in life scripts). For positive characteristics, earlier graduates remembered unpopular girls less favorably (perhaps using stereotypical scripts) than recent graduates remembered them (having greater access to episodic memories of individual girls). A smaller graduation time difference in the same direction resulted for average and popular girls. PMID- 18510140 TI - Administrative decision making: a stepwise method. AB - Today's health care organizations face tremendous challenges and fierce competition. These pressures impact the decisions that managers must execute on any given day, not to mention the ever-present constraints of time, personnel, competencies, and finances. The importance of making quality and informed decisions cannot be underestimated. Traditional decision making methods are inadequate for today's larger, more complex health care organizations and the rapidly changing health care environment. As a result, today's health care managers and their teams need new approaches to making decisions for their organizations. This article examines the managerial decision making process and offers a model that can be used as a decision making template to help managers successfully navigate the choppy health care seas. The administrative decision making model will enable health care managers and other key decision makers to avoid the common pitfalls of poor decision making and guide their organizations to success. PMID- 18510141 TI - Harassment: it's not (all) about sex! Part I: The evolving legal environment. AB - Policies prohibiting sexual harassment, although a good start, are not enough to protect health care employers from the risk of significant liability to an employee who experiences unlawful workplace harassment. The purpose of the first of this 2-part article is to help health care managers review what they already know and to update their knowledge of the legal environment associated with the ever-changing landscape of harassment in the workplace. PMID- 18510142 TI - Change management in health care. AB - This article introduces health care managers to the theories and philosophies of John Kotter and William Bridges, 2 leaders in the evolving field of change management. For Kotter, change has both an emotional and situational component, and methods for managing each are expressed in his 8-step model (developing urgency, building a guiding team, creating a vision, communicating for buy-in, enabling action, creating short-term wins, don't let up, and making it stick). Bridges deals with change at a more granular, individual level, suggesting that change within a health care organization means that individuals must transition from one identity to a new identity when they are involved in a process of change. According to Bridges, transitions occur in 3 steps: endings, the neutral zone, and beginnings. The major steps and important concepts within the models of each are addressed, and examples are provided to demonstrate how health care managers can actualize the models within their health care organizations. PMID- 18510143 TI - Training health care providers to be educators. AB - BACKGROUND: Students in the allied health field who have a positive clinical education experience often accept employment opportunities offered by their clinical site. Most allied health professionals are not trained to be educators. Professional development in the area of teaching could assist the professional to be a more effective teacher and therefore improve the clinical experience. PURPOSE: This study examined the professional development needs of clinical supervisors for a state university as they relate to being educators. METHODS: A survey was mailed to 520 clinical supervisors from an allied health program in a state university in Illinois. The survey addressed the clinical instructors'/preceptors' confidence level in their teaching skills and their interest in attending training on teaching strategies. FINDINGS: Results revealed that clinical supervisors in rural areas are less confident in their clinical teaching skills but are more confident in their ability to practice evidence-care based care and to work with their community compared with their urban and suburban counterparts. Respondents were also interested in a workshop or having access to a Web site on effective clinical teaching strategies. CONCLUSIONS: Clinical supervisors have a need and interest in improving their teaching skills. Improving clinical teaching skills could be addressed by providing a continuing education workshop and/or developing an academic Web site on effective teaching strategies. PMID- 18510144 TI - Awareness of the National Standards for Culturally and Linguistically Appropriate Services at an academic health center. AB - The study's objectives were to (a) determine the level of familiarity of faculty and students at an academic health center with the National Standards for Culturally and Linguistically Appropriate Services (CLAS), (b) identify faculty's and students' interest and preferred method of learning Medical Spanish, and (c) determine their aptitude for working with medical interpreters. A survey was developed, piloted, and sent via e-mail to all faculty members (n = 1,025) and students (n = 1,956) currently affiliated with or enrolled at the Medical College of Georgia. Reminder e-mails were sent after 2 weeks, and responses were accepted for 1 month. The total response rate for faculty members was 29% (300/1,025), and that for students was 44% (871/1,956). Nearly 22% of the responding faculty and 23% of the responding students reported that they were less than familiar with the National Standards for CLAS. Both faculty (46%) and students (70%) were willing to spend time learning Medical Spanish. Web-based instruction was the preferred educational delivery mode for those who completed the survey; however, 18% of faculty and 5% of students strongly disagreed with this point. When questioned about how often interpreters services are used, the rates for faculty and students ranged from 34% to 39%. These results suggest that a void exists in understanding the National Standards for CLAS and that there are varying levels of willingness to learn medical Spanish. PMID- 18510145 TI - Web 2.0: what a health care manager needs to know. AB - Web 2.0 is one of the latest buzzwords for an assortment of emerging technologies on the Web. Health care managers need to know the benefits and drawbacks of these technologies before integrating them into organizational operations. In this article, we first illustrate the differences in health care management style and workflow between the Web 1.0 and Web 2.0 technologies. After defining and explaining some of the representative technologies, we discuss the benefits of Web 2.0 in general and the reasons why a health care manager should know these trends. And lastly, we list some caveats that a health care manager should know before fully embracing the technologies. The keys are to align the technologies with the culture and workflow of the organization and have a clear policy on their usages. PMID- 18510146 TI - Hospital financial performance: does IT governance make a difference? AB - This study examined whether information technology (IT) governance, a term describing the decision authority and reporting structures of the chief information officer (CIO), is related to the financial performance of hospitals. The study was conducted using a combination of primary survey data regarding health care IT adoption and reporting structures of Florida acute care hospitals, with secondary data on hospital financial performance. Multiple regression models were used to evaluate the relationship of the 3 most commonly identified reporting structures. Outcome variables included measures of operating revenue and operating expense. All models controlled for overall IT adoption, ownership, membership in a hospital system, case mix, and hospital bed size. The results suggest that IT governance matters when it comes to hospital financial performance. Reporting to the chief financial officer brings positive outcomes; reporting to the chief executive officer has a mixed financial result; and reporting to the chief operating officer was not associated with discernible financial impact. PMID- 18510147 TI - Organizational characteristics associated with cultural and linguistic service provision within Alabama hospitals. AB - Like several states in the Southeast, Alabama is in the nascent stages of an increase in the population of foreign-born individuals for whom English is a second language. These individuals are also culturally different from the traditional southern population. Given the impact of culture and language on a person's service utilization, the introduction of new cultures may pose significant challenges for Alabama's health care providers if they are not prepared. The purpose of this project is to examine the organizational characteristics associated with the provision of culturally and linguistically appropriate services by Alabama hospitals. The data for the project come from a survey of all medical/surgical hospitals (N = 101). Fifty-nine surveys were returned, giving us a 58% response rate. The data were analyzed using correlations, analysis of variance, and logistic regression. Approximately 47% of the sample hospitals reported having a staff interpreter. Furthermore, hospitals that had staff interpreters did seem to be more aware of their community, which was reflected in their mission statements. In addition, directors who viewed their role as fulfilling the strategic plan accepted the task of providing staff interpreters. Thus, several hospitals in Alabama seemed to be ready to meet the cultural and language needs of their markets. PMID- 18510148 TI - Straight talk for health care managers: back to the basics of leadership. AB - Managers in the health care field should continually reflect on their leadership practices and the performance of individuals in their respective units, with an eye toward continuing improvement. A critical part of the reflective process is a review of the basics of leadership. This article presents information concerning 18 critical leadership topics and offers suggestions for improving performance. PMID- 18510149 TI - [Palpable purpura with severe pruritus]. PMID- 18510150 TI - [Irritable bowl or maldigestion: why is digestion not working properly?]. PMID- 18510151 TI - [Sauna clients are healthier. Sweating for long life]. PMID- 18510152 TI - [Altitude sickness in the airplane?]. PMID- 18510153 TI - [Symptoms without a finding. Real or "imagined" disease?]. PMID- 18510154 TI - [Presentation, Diagnosis and Management of fatigue in general practice]. PMID- 18510155 TI - [Recognition and management of panic disorder in primary care]. PMID- 18510156 TI - [Antibiotic therapy in the family doctor's office]. PMID- 18510157 TI - [Intensive blood sugar treatment in type 2 diabetics: no evidence for increased mortality in the ADVANCE study compared with ACCORD study]. PMID- 18510158 TI - Comparison between collar and cuffs and above elbow back slabs in the initial treatment of Gartland type I supracondylar humerus fractures. AB - We compared two commonly used methods of immobilization of Gartland type I supracondylar humeral fractures, with respect to pain control, use of analgesia and sleep interruption. Forty patients were included in the study, collar and cuff immobilization (group 1, n=20) and above elbow back slab immobilization (group 2, n=20). Diagnosis was made in the accident department and patients were immobilized (collar and cuff or back slab) according to the preference of the treating doctor. Patients were then reviewed in the next available fracture clinic where they were assessed. The Wong-Baker faces scale was used to measure pain. Patients immobilized with a collar and cuff had an average pain score of 7.2 compared with 3.4 for those immobilized with a back slab (P<0.0001). Children in the collar and cuff group used analgesia at regular intervals nearly four times more often than those in the back slab group (P=0.005), and 85% of children immobilized with a collar and cuff had interrupted or no sleep throughout the night following the injury (P=0.008) compared with 45% of children in the back slab group. We conclude that immobilization of Gartland type I fractures with an above elbow back slab provides better pain relief and is more comfortable for paediatric patients than collar and cuff immobilization. PMID- 18510159 TI - Spatial frame correction of anterior growth arrest of the proximal tibia: report of three cases. AB - We report three cases of anterior growth arrest of the tibia in adolescent boys. Two of the three cases had a clear history of trauma and although there was a history of trauma in the third case, the patient also had human leukocyte antigen B27 negative enthesitis that had not previously affected the knee. In all cases, a slowly deteriorating hyperextension of the knee developed that in the two posttraumatic cases was initially misinterpreted as a posterior cruciate ligament injury. Radiographs demonstrated an abnormal relationship between the tibial plateau and the long axis of the tibia on the lateral view--anatomical posterior proximal tibial angle. All patients were successfully treated by the Taylor spatial frame with correction of the anatomical posterior proximal tibial angle to normal, equalization of the legs and correction of any concomitant coronal deformity. Hyperextension of the knee developing after an injury in adolescents should raise the suspicion of anterior growth arrest prompting careful analysis of the lateral radiograph. PMID- 18510160 TI - Fracture dislocation of sacroiliac joint associated with triradiate cartilage injury in a child: a case report. AB - We report a very rare injury of a 8-year-old girl with sacroiliac fracture dislocation and triradiate cartilage separation. After the restoration of the sacroiliac joint by open means, reduction of the separated cartilage was seen. At 20 months follow-up, an osseous bridging at the triradiate cartilage and mild coxa valga deformity developed. We think that every child with serious sacroiliac joint injury should be evaluated for associated triradiate cartilage injury and followed to skeletal maturity for late complications such as acetabular dysplasia, hip subluxation and pelvic asymmetry. PMID- 18510161 TI - Iliac apophyseal displacement: an alternative in pediatric pelvic osteotomies. AB - The traditional surgical exposure for a Salter or Chiari pelvic osteotomy involves splitting the iliac apophysis to facilitate subperiosteal separation of the muscles of the inner and outer table of the ilium. With healing, the iliac crest frequently becomes broad and prominent, and the iliac wing hypoplastic. We addressed this issue by separating the whole iliac apophysis laterally at the junction of cartilage and bone and displacing it medially. The ilium was then exposed by subperiosteal dissection of the inner and outer table musculature. From February 1988 to June 2000, twenty-five pelvic osteotomies were performed utilizing this approach. Satisfactory exposure was achieved in each case. All osteotomies healed without iliac growth disturbances, leaving excellent cosmetic results. Resuturing the previously elevated external oblique abdominus over the iliac apophysis further improved contour and appearance, Iliac apophyseal displacement rather than splitting provided appropriate access and consistently good function and cosmesis. PMID- 18510162 TI - Arthrographically defined subchondral defects in Perthes' disease. AB - Since the original 1910 description of Perthes' disease, the aetiology and pathophysiology of this condition have remained elusive, and the treatment controversial. We found during arthrography that it has been possible to demonstrate a fluid-filled space between the ossified epiphysis of the femoral head and its overlying articular cartilage. This finding has not previously been documented in the literature, and we believe this mechanically vulnerable region may be subject to mechanical distortion, acting as a significant contributor to the evolving femoral head deformity seen in Perthes' disease. If this is so, treating this lesion could prevent further femoral head deformation. PMID- 18510163 TI - Ipsilateral teratologic high hip dislocation, femoral hypoplasia and below-knee hemimelia associated with contralateral fibular hemimelia: report of a case with functional result. AB - In this case report, we present a patient with right teratologic high hip dislocation, femoral hypoplasia and below-knee hemimelia associated with left fibular hemimelia. Combined open reduction, proximal femoral osteotomy and a Dega acetabuloplasty were performed in the right hip. Closed tibial wedge osteotomy and centralization of the foot with lateral release and Achilles tendon lengthening were performed for the left side. The patient was able to walk with her prosthesis successfully within the first six postoperative months. PMID- 18510164 TI - Video-assisted gastrocnemius-soleus and hamstring lengthening in cerebral palsy patients. AB - The aim of the study was to present the results of video-assisted fractional lengthening of the triceps surae muscle and the hamstrings in children with spastic cerebral palsy. In the period from September 2003 to December 2004, triceps surae muscle contractures were treated in 35 lower extremities (22 patients) and hamstring lengthening was performed in 12 knees (eight patients). The patients were between 4 and 10 years of age. Lengthening of the gastrocnemius soleus was sufficient for achieving 10 degrees dorsiflexion of the foot in 31 of the 35 extremities. The short-term follow-up, at least 1 year after operation, did not reveal any complications. The hamstring lengthening resulted in full correction in nine knees; one endoscopic procedure required conversion to open surgery owing to bleeding. In one case, incomplete sciatic nerve palsy developed. Video-assisted gastrocnemius-soleus recession as well as video-assisted lengthening of the hamstrings proved to be fully efficient in the group reported here. PMID- 18510165 TI - Congenital clubfoot with concomitant peroneal nerve palsy in children. AB - We investigated the prognosis and clinical importance of concomitant peroneal nerve palsy in congenital clubfoot. Six children (6 feet) with concomitant peroneal nerve dysfunction treated by various means and with no other related syndrome or neuromuscular disorder were studied. Their clinical outcomes and their parents' satisfaction with the outcome were assessed. Patients' average age at initial diagnosis was 5.1 months (range, 2 weeks to 13 months). The average length of follow-up was 67.4 months (range, 1-11 years). Nerve conduction velocity tests and electromyography were performed in six patients; serial cast correction, followed by application of an orthosis and periodic observation, was followed in two patients; and additional surgical correction was done in four patients whose deformity was not corrected by casting and an orthosis. None of the six patients recovered from peroneal nerve palsy and their parents reported a low level of satisfaction with the outcome of the treatment. It is important to detect the rare combination of peroneal nerve palsy and clubfoot, and to explain the poor prognosis to patients' parents before treatment. PMID- 18510166 TI - Solitary osteochondroma long-term follow-up. AB - A series of 113 osteochondromas were treated in our institution with a long-term follow-up. A retrospective study of the cases of solitary osteochondroma diagnosed and treated in our center from 1970 to 2002 was done. A diagnosis for clinical findings in 73% of the patients was made. The most frequent location was the distal femur. Six patients had a recurrence and in two patients the lesions became malignant and chondrosarcoma developed. Relapse of the exostosis is rare, occurring in an estimated 2% of the resections. The growth of an osteochondroma and/or the presence of pain in older patients suggest possible malignancy. PMID- 18510167 TI - The ulnius: a one-bone forearm in children. AB - Surgical creation of a one-bone forearm is appropriately utilized for a variety of underlying conditions. It functions best when the proximal ulna and distal radius are available for use. A variety of surgical techniques have been utilized. This report reviews the pediatric literature and presents four personal cases, each with a different underlying abnormality. The rotational position of forearm fusion is discussed. PMID- 18510168 TI - Os odontoideum in achondroplasia: a case report. AB - The neurological complication in achondroplasia has been focused on the foramen magnum stenosis. We report the combination of os odontoideum in a patient with achondroplasia. PMID- 18510169 TI - Metastatic renal cell carcinoma: recent advances and current therapeutic options. AB - For many years immuno(chemo)therapy has been the only therapeutic option for patients with metastatic renal cell carcinoma. Few patients, however, experienced long-term disease control and toxicity was considerable. Recent advances in understanding the biology and genetics of this malignancy have led to novel targeted therapeutic approaches. Since 2003, a multitude of new drugs have been developed and tested, with small molecule tyrosine kinase inhibitors, mammalian target of rapamycin inhibitors and monoclonal antibodies appearing to be the most promising agents. In the following, we give a concise overview on results of current trials in metastatic renal cell carcinoma published within 2007. Moreover, we will translate these results into therapeutic options and recommendations. PMID- 18510170 TI - Dihydroartemisinin induces apoptosis in human leukemia cells HL60 via downregulation of transferrin receptor expression. AB - Dihydroartemisinin (DHA), a water-soluble active metabolite of artemisinin derivatives, is the safest and most effective antimalarial analog of artemisinin. In the present investigation, we assessed the apoptotic effect of DHA on leukemia HL60 cells and its regulation of transferrin receptor (TfR). Cell growth inhibition was assessed by Trypan blue exclusive staining; the expression of caspase-3, Bcl-2, and Bax in HL60 cells was evaluated by Western blotting; DHA induced apoptosis was determined by AO/EB double staining, DNA fragmentation assay, and flow cytometric analysis; the expression of TfR in HL60 cells was examined by real-time PCR assays, Western blotting, and flow cytometric analysis. DHA could specifically reduce the mRNA and protein expression of TfR in HL60 cells, and the flow cytometric analysis presented the unity tendency that the TfR content decreased progressively in a dose-dependent manner. Consequently, DHA exhibited high anticancer activity in HL60 cells; MTT assay and growth inhibition assay showed that DHA could specifically inhibit the growth of HL60 cells in a dose-dependent (0.25-8 micromol/l) and time-dependent (12-72 h) manner. DHA induced DNA fragmentation also induced the activation of caspase-3 and influenced the expression of Bcl-2 and Bax. Taken together, these data from our study show that DHA can induce HL60 cell apoptosis via the effect of downregulation TfR expression resulting in an induction of apoptosis through the mitochondrial pathway, and it might be a potential antileukemia strategy for leukemia therapy. PMID- 18510171 TI - Multiple pathways are involved in drug resistance to doxorubicin in an osteosarcoma cell line. AB - Drug resistance continues to be a stumbling block in achieving a better cure rate in several cancers, including osteosarcoma. To understand this, we developed a doxorubicin drug-resistant osteosarcoma cell line (143B-DR-DOX). This cell line had an IC50 of 75 micromol/l compared with the parental 143B cell line's IC50 of 0.4 micromol/l. Using a 22000 70-mer oligomicroarray, gene expression studies were performed in four replicates. Data analysis was done using the TIGR Microarray suite. Seventy-four genes were found to be either upregulated (21) or downregulated (53). Real time quantitative-PCR was done on 21 genes, which confirmed the gene expression data for 11 genes. Choosing the significant fold change criteria of greater than 2-fold upregulation or downregulation, four genes including multidrug resistance 1, interleukin-8, Kruppel-like factor 2 and MGC4175 were found to be upregulated and seven genes including epidermal growth factor receptor-coamplified and overexpressed protein, uridine phosphorylase 1, a disintegrin and metalloproteinase domain 19, cytochrome C1, SEC, S-adenosyl homocysteine hydrolase and p53 were found to be downregulated. The data suggest that apart from the known gene alterations in doxorubicin resistance (multidrug resistance 1, topoisomerase IIbeta), others can also contribute to the drug resistance phenotype. The involvement of interleukin-8 and Kruppel-like factor 2 suggests that the peroxisome proliferator-activated receptors gamma pathway may also be involved in doxorubicin drug resistance in the 143B-DR-DOX cell line. PMID- 18510172 TI - High-dose methotrexate in adults with osteosarcoma: a population pharmacokinetics study and validation of a new limited sampling strategy. AB - Preoperative high-dose methotrexate (HD-MTX) with folinic acid (leucovorin) rescue is still a mainstay in the treatment of osteosarcoma. This anticancer agent is characterized by a narrow therapeutic index and wide interpatients variability. To ensure effective and safe administration of HD-MTX, we had earlier developed an adaptive-dosing schedule with a feedback strategy. In our institute, the MTX dosage was tailored according to individual pharmacokinetics parameters, determined in real time both from two blood samples (3.5 and 4.5 h) and from Bayesian population parameters. Up to 20 g of MTX was safely administered as 8-h infusions. Low MTX elimination rate has, however, been reported in 15-20% of the patients, and forecasting the MTX elimination phase and the management of leucovorin rescue is still a challenging issue in clinical oncology. This study aims at identifying the clinical or biological covariates related to impaired MTX clearance, and at validating a new limited sampling strategy (LSS), allowing for the accurate prediction of the MTX terminal elimination phase. This retrospective study was carried out on 49 patients (30 men, 19 women; mean age, 26.7 years) treated for osteosarcoma with HD-MTX. The population and individual pharmacokinetics parameters were computed, before the identification of the relevant covariates. Different LSSs were then tested, to predict accurately when the MTX plasma concentrations would drop below 0.2 micromol/l, the threshold associated with the end of the rescue of leucovorin with alkaline hydration. Two main covariates (creatinemia clearance and alanine aminotransferase) were correlated with MTX clearance. Conversely, the impact of body surface area on MTX pharmacokinetics was weak, suggesting that dosing schedules based on body surface area were inadequate and potentially hazardous. A new LSS predicting accurately when the MTX concentration would reach 0.2 micromol/l has been validated; blood samples are stopped as soon as the MTX concentration drops to 1 micromol/l. With this LSS, our retrospective study suggests that 60% of the patients would have left the hospital earlier than they actually did owing to a better forecasting of the MTX decrease, thus improving their quality of life while improving the cost-effectiveness for the institute. HD-MTX can be administered safely using an adaptive-dosing strategy with drug monitoring. Moreover, pharmacokinetic modeling permits the accurate forecasting of the MTX elimination profile, thus allowing for a better management of the postinfusion care of cancer patients treated with particularly high doses of this drug. PMID- 18510173 TI - Oral bioavailability of a novel paclitaxel formulation (Genetaxyl) administered with cyclosporin A in cancer patients. AB - The formulation excipient Cremophor EL (CrEL) is known to limit the absorption of oral paclitaxel given together with cyclosporin A. We hypothesized that the use of oral Genetaxyl, a paclitaxel formulation containing only 20% CrEL would have an improved oral bioavailability. Cohorts of six patients were treated with oral Genetaxyl at a dose of 60, 120, or 180 mg/m2 and 10 mg/kg of oral cyclosporin A in cycle 1. In cycle 2, patients received intravenous (i.v.) Genetaxyl (175 mg/m2, 3-h infusion). Three additional patients received one dose of generic i.v. paclitaxel (Genaxol, containing 50% CrEL; 175mg/m2, 3-h infusion). The median area under the plasma concentration-time curve (AUC) and peak concentration of total paclitaxel following i.v. Genetaxyl were lower than those for i.v. Genaxol, as a result of significantly increased clearance (P = 0.017), and the AUC ratio for unbound to total paclitaxel for i.v. Genetaxyl was about two times higher than that for i.v. Genaxol (P = 0.0077). After oral administration of Genetaxyl at doses of 60, 120, and 180 mg/m2, the median total paclitaxel AUCs were 1.29, 1.60, and 1.85 microg x h/ml, respectively, suggesting a less than proportional increase in systemic exposure with increasing doses. The corresponding median values for the apparent bioavailability of oral Genetaxyl were similar when compared with i.v. Genetaxyl, when calculated either on the basis of data for total paclitaxel (30.1%) or unbound paclitaxel (30.6%). PMID- 18510174 TI - Phase II study of oxaliplatin in combination with continuous infusion of 5 fluorouracil/leucovorin as first-line chemotherapy in patients with advanced gastric cancer. AB - This study was designed to determine the efficacy and safety of biweekly oxaliplatin in combination with infusional 5-fluouracil (5-FU) and leucovorin in patients with advanced gastric cancer (AGC). Fifty-five eligible patients with measurable or assessable M/AGC (median age 62 and 90% of patients presented with metastasis) received oxaliplatin (85 mg/m2) intravenous infusion for 2 h, followed by intravenous infusion of 5-FU (3000 mg/m2) and leucovorin (100 mg/m2) for 46 h every 14 days until the patient's disease was either in progression, unacceptable toxicity, patient's withdrawal or the investigators' decision to discontinue treatment. Of the 55 enrolled patients, 48 were evaluable for response. Three patients (5.4%) showed complete remission and 20 patients (36.4%) achieved partial response. The overall response rate was 47.9%. Nineteen patients (34.5%) had stable disease and six patients (10.9%) showed progressive disease. The median time to progression was 5.6 months and the median overall survival was 10.8 months. Grade 3/4 toxicities included leucopenia (12.7%), thrombocytopenia (5.4%), diarrhoea (3.6%) and vomiting (9.1%). Peripheral neuropathy was noted in 61.8% of the patients (grade 1/2: 54.5%; grade 3: 7.3%). Our study confirmed that the combination of oxaliplatin and continuous infusion of 5-FU/leucoverin without bolus 5-FU as first-line chemotherapy is active for patients with AGC and relatively safe with lower haematological toxicity. PMID- 18510175 TI - Phase I trial of oral S-1 plus gemcitabine in elderly patients with nonsmall cell lung cancer. AB - We conducted a phase I study to determine the maximum tolerated dose, the recommended dose and the safety profile of S-1 and gemcitabine combination regimen in the treatment of elderly patients (> or = 70 years) with advanced nonsmall cell lung cancer (NSCLC). Chemotherapy-naive patients with advanced NSCLC were treated with S-1 and gemcitabine. S-1 was administered orally twice daily for 14 days and gemcitabine on days 1 and 15 of each cycle, and this was repeated every 4 weeks. Doses of each drug were planned as follows: level 1, 800/60; level 2, 1000/60; level 3, 1000/70; and level 4, 1000/80 [gemcitabine (mg/m2/ day)/S-1 (mg/m2/day)]. The dose-limiting toxicity (DLT) of the regimen was assessed during the first chemotherapy cycle. Sixteen patients were enrolled in this study. The main grade 3 toxicities observed during the first cycle were neutropenia (43.7%), leukopenia (18.7%), and hyperglycemia. One of six patients in level 3 had DLT. Although no patients in level 4 experienced DLT, this level was considered the maximum tolerated dose. Level 4 was selected as the recommended dose. Objective responses were seen in four patients (response rate, 42.9%). The combination of S-1 plus gemcitabine is a feasible and well-tolerated regimen for the treatment of elderly patients with advanced NSCLC. PMID- 18510176 TI - An original administration of ifosfamide given once every other week: a clinical and pharmacological study. AB - Ifosfamide (IFOS) is a bifunctional alkylator with a wide spectrum of activity in solid tumors and has an autoinductive liver metabolism through P450 cytochromes. Autoinduction might permit a better therapeutic index for combination therapy. A phase I trial was investigated with interpatient dose escalation of a single dose of IFOS given every 2 weeks in advanced solid tumor patients. IFOS, its dechloroethylated and active 4-hydroxy metabolites, were measured at cycles 1 and 2 at the end of infusion, 2 and 5 h later, using gas chromatography. IFOS elimination was considered as following monocompartimental model kinetics. The results of 20 patients from January 2004 to June 2006 were included. The median of previous chemotherapies was 2 (0-5). The primary tumor was most often ovarian (5), peritoneal (3), sarcoma (2), melanoma (2) or miscellaneous (8). Ten patients received 2.5 g/m2 and the other 10 patients received 3 g/m2. A total of 79 cycles were evaluable for toxicity. The median number of cycles was 4 (1-8). No grade 3 4 toxicity, no alopecia at first dose level and no toxicity-related fatal events were noted. One objective response was noted in a pancreatic cancer patient and one sustained CA125 decline in a heavily pretreated ovarian cancer patient. A slight (7-10%) but reproducible decrease of areas under the curve was detectable at cycle 2, at both dose levels, related to autoinductive metabolism. Intraindividual variations (large SD) were noticed for each pharmacokinetic parameter. A patient-dependent autoinduction of IFOS metabolism was detected rather than a slight nondose-dependent autoinduction. The toxicity profile allows the development of bi-weekly IFOS-based combination therapies. PMID- 18510177 TI - Mitomycin C plus S-1 as second-line therapy in patients with advanced gastric cancer: a noncomparative phase II study. AB - S-1 is an oral fluoropyrimidine consisting of the 5-fluorouracil prodrug tegafur combined with two modulating substances, gimeracil and potassium oxonate. On the basis of the potential additive effect between mitomycin C (MMC) and 5 fluorouracil as a continuous infusion, we conducted a phase II study to assess the efficacy and tolerability of the combination of S-1 and MMC as second-line chemotherapy for advanced gastric cancer (AGC). Patients with measurable AGC, progressive after one prior chemotherapy for metastatic disease, received MMC (7 mg/m2) on day 1 and S-1 (40 mg/m2) twice daily as an intermittent regimen of 4 weeks of treatment followed by a 2-week rest. Treatment was repeated every 6 weeks. The primary objective was the response rate. For 43 patients registered, 42 patients were treated with MMC plus S-1. A total of 121 chemotherapy cycles were delivered (median: 2; range: 1-6). The patients' median age was 53 years (range: 31-75) and nine (21%) had an Eastern Cooperative Oncology Group performance status of 2. In an intent-to-treat analysis, nine patients (21%) achieved an objective response, which was maintained for 4.1 months. The median progression-free and overall survivals were 3.4 months (95% confidence interval: 2.3-4.5) and 8.0 months (95% confidence interval: 6.1-9.9), respectively. Although fatigue was the most frequently encountered toxicity safety profiles were generally predictable and manageable. One patient developed hemolytic anemia, which was resolved spontaneously. Grade > or = 2 hand-foot syndrome was observed in only three patients. Second-line chemotherapy with MMC and S-1 is an active and tolerable regimen for AGC patients with good performance status. PMID- 18510178 TI - A multicenter study of gemcitabine-containing regimen in relapsed or refractory Hodgkin's lymphoma patients. AB - The aim of this study was to assess the efficacy of a gemcitabine-containing regimen in pretreated Hodgkin's lymphoma (HL) patients. Relapsed or refractory HL patients treated with gemcitabine, used alone or in combination with other cytotoxic agents, were retrospectively reviewed. Fifty-five patients were included in the study. Initial characteristics before gemcitabine administration were: Ann Arbor stage III-IV: 84%; International Prognostic Score less than 3 in 18/39 cases (46%); 31 primary refractory patients at the end of first-line therapy (56%); median number of previous chemotherapy regimens of 3. Twenty-nine patients received gemcitabine alone with a median maximal dose of 900 mg/m2 per injection (range: 300-1500 mg/m2). Gemcitabine was administered at a maximal dose of 1000 mg/m2 per injection (range: 650-1250) in combination with vinorelbine in 10 patients, oxaliplatin in 13 patients, and other drugs in three patients, with a median of six injections (range: 1-18). Reported toxicity was mainly hematologic. Overall response rate was 20% with 11% of complete remission. On univariate analysis, two adverse factors at progression were significant for response to gemcitabine-based regimen: stage III-IV disease and hemoglobin level was less than 10.5 g/dl. This study demonstrated the limited efficacy of gemcitabine-containing regimen in heavily pretreated HL patients. PMID- 18510179 TI - Predictive value of serum anti-p53 antibodies, carcino-embryonic antigen, carbohydrate antigen 15-3, estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2 in taxane-based and anthracycline-based neoadjuvant chemotherapy in locally advanced breast cancer patients. AB - Breast carcinoma is the most common malignancy in Chinese women. The purpose of this study is to evaluate the predictive value of serum anti-p53 antibodies (p53 Abs), carcino-embryonic antigen (CEA), carbohydrate antigen (CA) 15-3, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor (HER)-2 in taxane-based and anthracycline-based neoadjuvant chemotherapy (NAC). Sixty-eight patients with locally advanced breast carcinoma were included. Thirty-two were treated with taxane (the taxane group) and 36 with anthracycline (the anthracycline group). The standard dosage of docetaxel was 100 mg/m2 (day 1) and those of cyclophosphamide, adriamycin and 5-flurouracil were 500 mg/m2 (day 1 8), 40 mg/m2 (day 1) and 500 mg/m2 (day 1-8), respectively. The p53 Abs were detected by enzyme-linked immunosorbent assay; CEA and CA15-3 were detected by Elecsys 2010 Disc System; ER, PR and HER-2 were detected by immunohistochemistry staining. The biomarkers p53 Abs, CEA and CA15-3 were detected in serum samples, and the immunohistochemistry staining for ER, PR and HER-2 was performed in tumor samples before and after NAC. The expression of p53 Abs was significantly reduced by taxane (P = 0.006). The serum CEA and CA15-3 levels were significantly affected by both taxane (P = 0.004 and P = 0.008) and anthracycline (P = 0.002 and P = 0.000) drugs. HER-2-negative status (pre-neoadjuvant) was correlated with a high objective response rate (OR) in both taxane-based and anthracycline-based chemotherapy (P = 0.022 and P = 0.025), whereas p53 Ab-negative status (pre neoadjuvant) was correlated with high OR rate in anthracycline-based chemotherapy (P = 0.039). This study shows that the serum p53 Ab level is easily changed by taxane. CEA and CA15-3 levels are easily changed by taxane and anthracycline. The p53 Ab-negative patients may predict a high clinical OR rate in anthracycline based NAC. HER-2-negative may predict a high OR in both taxane-based and anthracycline-based NAC. PMID- 18510180 TI - Severe bladder contracture leading to cystectomy after intravesical mitoxantrone chemotherapy. AB - Since the early 1990s, mitoxantrone has been used as a chemotherapeutic agent for adjuvant intravesical treatment following transurethral resection of superficial transitional cell carcinomas of the bladder. Although its efficacy as adjuvant intravesical therapy remains questionable and its use has not gained wide acceptance, the safety profile of the drug has been reported as favorable. We report the first case of mitoxantrone-induced severe bladder contracture leading to a completely nonfunctional bladder after intravesical administration of the drug. Cystectomy and urinary diversion were the final consequences for the patient. PMID- 18510181 TI - The use of beta-blockers to decrease adverse perioperative cardiac events. AB - As the population ages, more surgeries are performed on patients with increased risk factors. Many of these surgeries will result in complications and mortality. Myocardial ischemia is often associated with cardiac complications in the perioperative patient. Unfortunately, an episode of myocardial ischemia will result in a 70% mortality rate. Perioperative patients are complex because of their atypical presentation, incisional pain, and the effects of analgesia. The critical care nurse must be able to determine a patient's risk factors and surgical risks and then take measures to ensure that the patient receives adequate treatment both before and after surgery. beta-Blockers may decrease a patient's risk of developing myocardial ischemia, yet they are often underused. A multidisciplinary approach is needed to ensure the proper use of beta-blockers in an effort to reduce cardiac complications in the perioperative elderly patient. PMID- 18510182 TI - Adverse hemodynamic effects of lateral rotation during mechanical ventilation. AB - Turning critically ill, mechanically ventilated patients every 2 hours is a fundamental nursing intervention to reduce the negative impact of prolonged immobility from preventable pulmonary complications such as ventilator-associated pneumonia and atelectasis. Unfortunately, when coupled with positive pressure ventilation, the benefits of turning may come at the expense of cardiovascular function. Clinicians should closely monitor the hemodynamic response to turning mechanically ventilated patients, and if compromise is observed, the degree and duration of compromise may provide guidance to the appropriate intervention. PMID- 18510183 TI - The peer review process. AB - Scholarly journals use a peer review process when deciding whether to accept a manuscript for publication. This article will describe the peer review process, identify the qualities of a good peer review, and provide suggestions on how to become a peer reviewer. PMID- 18510184 TI - Your role in informed consent. Part 1. AB - As critical care nurses, we all participate in the informed consent process, whether for a routine procedure or surgery or for a research study or clinical trial. This first article of a 2-part series discusses the critical care nurse's role concerning informed consent for surgery or other invasive procedures, including nursing procedures. Part 2 will discuss the critical care nurse's role concerning informed consent for clinical trials. PMID- 18510185 TI - Barriers and facilitators to the use of evidence-based best practices. AB - The continued use of healthcare interventions without an evidence base increases healthcare costs without positively impacting patient care outcomes. Reports disseminated by bodies such as the Institute of Medicine and initiatives such as the Institute for Health Care Improvement's 5 Million Lives Campaign have increased emphasis on improving outcomes. Results of a descriptive correlational study indicated that 64% of the nurses surveyed read 7 or more specialty journals, 53% read 1 or more general nursing journal, 20% did not regularly read any professional journal, and none of the nurses surveyed read a journal that was primarily dedicated to the publication of original research. Almost half of the nurses indicated that the hospital library was the nearest location to conduct searches, and 34% indicated that they did not know what literature-searching capabilities were available to them. Although knowledge in itself is not sufficient for behavior change, it is an essential prerequisite. Regular reading of journals either through personal subscriptions or access through facility libraries can encourage the adoption of new evidence through lifelong learning. Modeling and skill building in use of readily available Internet resources can serve as a mechanism to increase awareness of and skill in accessing current information. Evidence-based changes can then be empirically examined, implemented, and evaluated in examining nursing's contribution to the daily operation of the healthcare organization. PMID- 18510186 TI - Scientific misconduct. PMID- 18510187 TI - Quality of nursing home care in Cyprus: are elder residents content with their treatment? AB - Responding to a conspicuous dearth of knowledge on the quality of elder services in Cyprus, this study ventured to document Cypriot elders' feelings and experiences with nursing home care in Cyprus. Explicitly, four different types of nursing homes were called on (a governmental, a community-run, a faith-based, and a private one) to interview residents (n = 73; a response rate of 100%). Results suggest that Cypriot elders are clearly content with the level of primary care they receive in nursing homes, as mirrored in the quality of nutrition, medical treatment, staff professionalism, and sanitation of nursing home environment. However, the preponderance of residents feels loneliness and denial of essential entertainment opportunities in these institutions and a consequential motivational depletion. Finding implications for domestic and international policy, social work practice, and future research are explored. PMID- 18510188 TI - Working with advanced dementia patients in a day care setting. AB - Alzheimer's disease and most other causes of dementia are regressive by nature. As such one can expect patients with such types of mental impairment to gradually decline in function and ability to participate in day care activities. This paper attempts to show that with the right kind of orientation, staff can "tune into" the more advanced dementia patients, find the key to their personal needs, desires and remaining abilities and design a program that allows them not only to continue to participate in a social and therapeutic framework, but also to gain some meaningful human contact and quality of life despite their cognitive deterioration. PMID- 18510189 TI - A measure of English acculturation stress and its relationships with psychological and physical health status in a sample of elderly Russian immigrants. AB - This study aims to evaluate the factor structure and criterion validity of the English Language Acculturation Stress (ELAS) scale in a community-based sample of elderly Russian immigrants. The sample consists of 300 Russian immigrants with an equal number of females and males, from 59 to 93 years of age (Mean = 73.26, SD = 7.271). The majority of the respondents were married (72%) at the time the interviews were conducted. On average, the respondents have lived in the U.S. for about 7 years (SD = 3.00). We performed both exploratory and confirmatory factor analyses to ascertain the factor structure or conceptual dimensions of the 11 item ELAS scale. The findings revealed that this 11-item scale encompasses three dimensions: Basic English skills, survival English skills, and social involvement English skills. Correlation analyses of the scale with depression, physical health status, and length of residence suggest that the ELAS has good criterion validity and potential as a screening instrument of language acculturation stress for elderly Russian immigrants. Implications for research and practice are discussed. PMID- 18510190 TI - Family councils in nursing facilities: strategies for effective participation. AB - This article provides an overview of the role of family councils. The contributions of the three groups that play a significant role in the development and support of family councils, relatives and friends of the facility's residents, facility administration and staff, and local advocacy organizations, are discussed. The role of the staff advisor, generally the facility social worker, is a major key to a successful council. Barriers that impede family council success are identified, and strategies that improve effectiveness are presented. Family councils play an important part in improving the quality of lives of long-term care residents. PMID- 18510191 TI - The parameters of prejudice: knowledge of ethics and age bias. AB - This study examined two matters pertinent to social work practice: professional ethics and age bias among practicing social workers. Because social work ethics demand competent practice within one's area of proficiency, and because bias toward any segment of social work clientele impedes competent practice, prejudicial attitudes toward older people are problematic. This study found that age bias exists among practicing social workers (N = 367), with no discernible association between knowledge of professional ethical standards and age bias. The findings suggest a subtle and pervasive bias associated with work with older people. Positive bias was more prevalent than negative bias. PMID- 18510192 TI - A look at a community coming together to meet the needs of older adults: an evaluation of the neighbors Helping Neighbors program. AB - The purpose of this study was to evaluate the effectiveness of the Neighbors Helping Neighbors program. The study included surveys of 49 community-residing older adults and 26 community volunteers. Results showed that older adults perceived their quality of life to have improved after receiving social and environmental services; volunteers felt that their contributions to the program had made a significant difference in their community. This exploratory, descriptive study is only a beginning effort, but it holds great promise for suggesting ways to address the needs of the burgeoning aging population in our society. PMID- 18510193 TI - Videos: Where do they fit in an aging infused social work curriculum? AB - As technology progresses, college instructors are presented with the availability of new and exciting pedagogical methods. Though the use of videos is not new, their use is becoming increasingly simplified and relevant to popular culture. This conceptual paper presents a theoretical rationale for the use of videos as a teaching and learning tool in the infusion of aging content into the social work curriculum, provides in-class strategies with a case example, and discusses the use of videos outside of class. PMID- 18510194 TI - Private prayer among Alzheimer's caregivers: mediating burden and resiliency. AB - This study examined whether the coping method of private prayer served as a protective factor of resiliency among a sample (N = 304) of Alzheimer's caregivers. Participants in caregiver support groups completed questionnaires that assessed a number of constructs, including caregiving burden; prayer frequency; use of private prayer as a means of coping; and perceived resiliency. The sample averaged a moderate level of burden and a great extent of prayer usage. Caregiving burden had positively affected the extent of prayer usage and negatively influenced perceived resiliency. Findings from hierarchical regression analysis showed that caregiving burden and private prayer significantly influenced variation in perceived resiliency scores. Results from a regression equation series and path analysis provided support for prayer as a mediator between burden and perceived resiliency. Implications for social work practice and education are discussed. PMID- 18510195 TI - BJ Spitler: geriatric care management pioneer. AB - The late BJ Curry Spitler pioneered a new geriatric care management business model that combined care management with home health services and emphasized high levels of training and professionalism for all members of the caregiving team. After working with BJ to write a marketing plan emphasizing quality, I learned to appreciate what she did when my mother needed "assisted living at home" services from the firm. Thanks to BJ Curry Spitler and her vision of helping seniors age in place, more elders are able to remain independent in their own homes with dignity and safety, and peace of mind for their families. PMID- 18510196 TI - Home health and informal care utilization and costs over time in Alzheimer's disease. AB - OBJECTIVES: To (1) compare home health and informal (unpaid) services utilization among patients with Alzheimer's disease (AD), (2) examine longitudinal changes in services use, and (3) estimate possible interdependence of home health and informal care utilization. METHODS: The sample is drawn from the Predictors Study, a large, multicenter cohort of patients with probable AD, prospectively followed annually for up to 7 years in three university-based AD centers. Bivariate probit models estimated the effects of patient characteristics on home health and informal care utilization. RESULTS: A large majority of the patients (80.6%) received informal care with a smaller proportion (18.6%) receiving home health services. Home health services utilization increased from 9.9% at baseline to 34.5% in year 4. Among users, number of days that services were provided in three-month recall increased from 21.9 to 56 days over time. Home health services utilization was significantly associated with function, depressive symptoms, being female, and not living with a spouse. Informal care utilization was significantly associated with cognition, function, comorbidities, and living with a spouse or child. CONCLUSIONS: Home health and informal care utilization relate differently to patient characteristics. Utilization of home health care or informal care was not influenced by utilization of the other. PMID- 18510197 TI - Enhancing health care communication skills: preliminary evaluation of a curriculum for family caregivers. AB - The Communicating Effectively with Health Care Professionals (CE) workshop curriculum is designed for family caregivers to encourage caregiver empowerment, effective health care communication, and advocacy in medical care contexts with the goal of promoting positive health outcomes for care recipients. This mixed method study employed a cross-sectional quantitative mail survey (N = 51) and semistructured qualitative telephone interviews (N = 14) to examine the effectiveness of the curriculum in promoting self-reported changes in caregiver attitudes and communication behavior. Respondents reported increased assertiveness in medical encounters, feelings of empowerment, and preparation and organization of medical information for their care recipients as a result of workshop participation. PMID- 18510198 TI - Barriers to workers' compensation and medical care for injured Personal Assistance Services workers. AB - OBJECTIVES: We documented barriers to workers' compensation and injury-related medical care faced by homecare or Personal Assistance Services (PAS) workers. We explored differences between independent providers and agency-employed workers. METHODS: We conducted in-depth, semi-structured interviews with a diverse sample of 38 injured workers. Participants were primarily female and racial-ethnic minorities. RESULTS: Most participants (82%) were independent providers. Common barriers to reporting injury included commitments to clients and financial pressure. Unlike agency employees, many independent providers knew little about workers' compensation eligibility and injury reporting procedures, and frequently were given "the runaround" by the social service bureaucracy when they attempted to report injury and access injury-related medical care. Among independent providers, delays in filing a claim and receiving timely medical attention were common. CONCLUSIONS: The lack of a traditional employment infrastructure has important implications for vulnerable workers' health and the sustainability of consumer-directed PAS programs. We provide recommendations for improving workers' access to workers' compensation and injury-related medical care. PMID- 18510199 TI - The relationship between agency characteristics and quality of home care. AB - BACKGROUND: This project assessed the relationship between home care quality indicators (HCQIs) and agency characteristics. METHODS: Twelve agencies completed a mailed survey on a variety of characteristics, including size of their caseload and for-profit (FP) status of contracted service providers. The HCQIs were derived from standardized assessments completed voluntarily for home care clients in Ontario and in Manitoba, Canada. RESULTS: The average caseload was 121.3 clients per case manager, and over 40% of nursing, personal support and therapy providers were considered FP. For individual HCQIs, few correlations were statistically significant. An overall summary measure of quality was correlated with the size of the population served (r = -0.80; p < 0.05) and the number of clients per case manager (r = -0.56; p < 0.1). CONCLUSION: These data represent unique information on home care quality and organizational characteristics in Canada. The question remains as to how best to use HCQI data to inform practice in an era of limited resources and increasing caseloads. PMID- 18510200 TI - Food-coping strategy index applied to a community of farm-worker households in South Africa. AB - BACKGROUND: In South Africa, households living in informal urban settlements, in rural areas, and on commercial farms experience various levels of dietary variety, food intake, and household hunger. Low incomes, poor food production and availability, and low spending power characterize these households. Households employ various food-coping strategies to alleviate food stress or poor food availability. OBJECTIVE: To apply an existing food-coping strategy (FCS) index to assess household hunger and its usefulness in identifying the level of food stress and the patterns of food coping in farm-worker households. METHODS: A cross-sectional survey was conducted. Data were gathered from women (18 to 57 years of age) responsible for food provision in a small farm-worker community in Fouriesburg, South Africa. A structured food-coping questionnaire and a standardized FCS index were used to gather data. RESULTS: The two most common FCS used were relying on cheaper food (chicken feet, diluted soya-mince soup) or less preferred food (meat bones) and employing food-seeking strategies (gathering wild foods), followed by consumption of seed stock (maize) and reduced portion sizes (protein foods and side dishes), resulting in starch-based diets of poor variety. Seasonal strategies varied according to the level of food stress experienced. Patterns of food coping were identified. CONCLUSIONS: Negative FCS (limiting food choices, only consuming starchy staples) may cause poor health status. The FCS index was effectively used to assess farm-worker household food-coping behavior (early, clear signals of the level of food distress). These results could be used to allocate appropriate food aid (type of food) and to design nutrition education programs focused on positive FCS (food gathering or bartering) in a particular community to prevent suboptimal nutritional status. PMID- 18510201 TI - Infant-feeding practices and beliefs about complementary feeding among low-income Brazilian mothers: a qualitative study. AB - BACKGROUND: Breastfeeding and weaning practices are important determinants of growth and development not only in infancy but also later in life. OBJECTIVE: To describe infant-feeding practices and beliefs about complementary feeding among low-income Brazilian mothers. METHODS: Qualitative methods included focus group discussions with low-income mothers enrolled in a Family Health/Community Health Workers program in Ceara, Northeast Brazil. RESULTS: Breastfeeding is widely practiced in this area, and overall, mothers are knowledgeable about the benefits of breastfeeding for their infants and themselves. Practices of prolonged breastfeeding and delayed supplementation of infants with semisolid foods emerged as a problem among very poor women. In addition, the results showed common problems related to complementary feeding practices, such as the early introduction of solid foods and the use of expensive commercial cereals and formula for weaning. Cultural factors and taboos appeared to have an important influence on mothers' infant-feeding practices and eating patterns of their children. CONCLUSIONS: The results have implications for the design of breastfeeding promotion and interventions to improve complementary feeding. Improvements of the national Food Grant Program are also suggested, which are needed by low-income mothers to improve their infant-feeding practices. PMID- 18510202 TI - Dietary intake and clinical response of hospitalized patients with acute diarrhea. AB - BACKGROUND: Diarrhea and malnutrition remain major health problems among children of developing countries. During diarrhea, the patient's dietary intake and absorption of nutrients are reduced while nutritional requirements are increased. OBJECTIVE: To determine the relationship between food intake and clinical response during the hospital stay of patients with acute diarrhea. METHODS: A hospital-based longitudinal study was conducted in 118 patients with acute diarrhea aged 6 to 59 months who required treatment for at least 3 days in the in patient ward in Dhaka Hospital of the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B). Daily food intake was measured and anthropometric measurements were taken to assess nutritional status. Daily stool weight and clinical records were collected. The data were analyzed with SPSS/PC+, version 10, and EPI STAT, version 3.2.2. RESULTS: The duration of diarrhea was 50% greater in patients with lower energy intake (less than 50% of the recommended dietary allowance [RDA]) than in those with higher energy intake (6 vs. 4 days, p = <.001). Patients with lower energy intake had 22% greater stool output than those with higher energy intake (122.65 vs. 100.37 mL/kg body weight/day, p = .04). Among patients with lower energy intake, the weight-for-age and weight-for-height z-scores (WAZ and WHZ) at discharge from the hospital were higher than those at admission (-3.53 +/- 1.25 vs. -3.67 +/- 1.31 and 1.95 +/- 1.23 vs. -2.14 +/- 1.22, respectively; p = .001 for both comparisons), but these scores did not differ at admission and discharge among patients with higher energy intake. The Kaplan-Meier survival function showed that 80% of well nourished children (WAZ > or = -2), as compared with 58% of malnourished children (WAZ < -2), recovered by the 4th day of treatment (p < .01). The length of the recovery period was related negatively with total energy intake (p = < .001) and mid-upper-arm circumference (p = .004) and positively with stool weight. CONCLUSIONS: Food intake was reduced in the hospitalized children because of severe illness. Patients with lower energy intake as a percentage of RDA had delayed clinical recovery and higher stool output. PMID- 18510203 TI - Impact of orphanhood on underweight prevalence in sub-Saharan Africa. AB - BACKGROUND: In Africa, approximately 25 million people live with HIV/AIDS and 12 million children are orphaned. Although evidence indicates that orphans risk losing opportunities for adequate education, health care, and future employment, the immediate effects of orphanhood on child nutritional status remain poorly understood. OBJECTIVE: This paper assesses the nutritional impact of orphanhood, with particular emphasis on taking account of various factors potentially confounding or masking these impacts. METHODS: Child anthropometry and orphan status were examined in 23 Multiple Indicator Cluster Surveys and Demographic and Health Surveys throughout sub-Saharan Africa, which were subsequently merged into larger, region-specific datasets (East, West, and Southern Africa). To compare orphans and nonorphans, linear regression and probit models were developed, taking account of orphan status and type, presence of a surviving parent in the household, household structure, child age and sex, urban versus rural residence, and current wealth status. RESULTS: Few differences emerged between orphans and nonorphans in controlled and uncontrolled comparisons, regardless of orphan type, presence of surviving parent, or household structure. Age differentials did confound nutritional comparisons, although in the counterintuitive direction, with orphans (who were 8 months older on average) becoming less malnourished when age differences were taken into account. Wealth did appear to be associated with orphanhood status, although it did not significantly confound nutritional comparisons. CONCLUSIONS: Orphans were not consistently more malnourished than nonorphans, even when potential confounding variables were examined. Since household wealth status is likely to change after becoming affected by HIV ruling out wealth as a potential confounder would require more detailed, prospective studies. PMID- 18510204 TI - Determination of the leading central obesity index among cardiovascular risk factors in Iranian women. AB - BACKGROUND: It is unknown whether the waist circumference (WC) or the waist-to hip ratio (WHR) is a better predictor of cardiovascular risk factors at different ages. OBJECTIVE: To compare WC and WHR as predictors of cardiovascular risk factors and to determine the prevalence of some cardiovascular risk factors in overweight and obese adult women at different ages. METHODS: In this clinical cross-sectional study, 714 overweight and obese women aged 20 to 70 years who were referred to two nutrition clinics in Sistan and Baluchestan province, Islamic Republic of Iran, were studied. The subjects were classified into three groups, 20 to < 35, 35 to < 50, and > or = 50 years of age. Anthropometric indices were measured according to the standard protocol. Total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and TC/HDL-C ratios were enzymatically determined. RESULTS: Older subjects (> or = 50 years old) had significantly higher values of body mass index (BMI), WC, TC, TG, and LDL-C than those in the two younger age classes. The prevalence rates of obesity, high WC, high WHR, high TC, high TG, high LDL-C, and high TC/ HDL-C ratios were higher in the older subjects. After adjustment for age and BMI, multiple linear regression showed that WC was significantly related to TC and TG in the 20- to < 35-year-old group and to TG in the 35- to < 50-year-old group. In the older participants, WHR was significantly related to TG. CONCLUSIONS: The prevalence of cardiovascular risk factors increases with age. In clinical practice, WC is a better index for predicting some cardiovascular risk factors in younger and middle-aged women; however, for older women, WHR is better. PMID- 18510205 TI - Nutritional status and its correlates in Equatorial Guinean preschool children: results from a nationally representative survey. AB - BACKGROUND: In Equatorial Guinea, as a result of the recent growth of the oil industry, there is an opportunity to address important public health problems through public and private initiatives. To propose effective nutrition and public health strategies, it is important first to have reliable information on the nutritional status of the population and the underlying factors affecting it. OBJECTIVE: To assess the nutritional status and the prevalence of anemia among Equatoguinean children in a nationally representative sample and to identify the risk factors associated with the nutritional problems detected. METHODS: The study was a cross-sectional survey using a multistaged, stratified, cluster selected sample. The survey included a sociodemographic, health, and dietary questionnaire and measurement of hematocrit and anthropometric features, from which nutritional indicators based on the National Center for Health Statistics (NCHS) reference and the World Health Organization (WHO) standards were calculated. Logistic regression models were used for the multivariate analysis. A total of 552 children aged 0 to 60 months were surveyed. RESULTS: The overall prevalence of stunting (< -2 height-for-age z-scores [HAZ]) was 29.7% based on the NCHS reference and 35.2% based on WHO standards; the risk factors associated with stunting were age (p < .0001), low socioeconomic status (p = .01), and fishing by a member of the household (p = .003) The prevalence of mild anemia (hemoglobin < 110 g/L) was 69.3%, and that of moderate or severe anemia (hemoglobin < 80 g/L) was 8.3%. The only significant risk factor associated with moderate to severe anemia was low household socioeducational level (p = .01). CONCLUSIONS: Stunting and anemia are public health problems in Equatorial Guinea. Integrated strategies, including fighting poverty and improving maternal education, should be undertaken. PMID- 18510206 TI - Relationship between paternal involvement and child malnutrition in a rural area of Vietnam. AB - BACKGROUND: Malnutrition is a public health problem in Vietnam. Child health and the status of women have been targets for various health programs in the country. In general, reports in the literature suggest that care is positively correlated with positive nutritional status of children. In the household, the father is considered a resource for care. However, the role of paternal care in health programs has not received the attention it deserves. OBJECTIVE: To identify associations between the involvement of fathers in child care and housework and the nutritional status of children under 3 years of age. METHODS: This cross sectional study was based on a random sample of 547 children under 3 years of age from intact families and their biological parents. The main outcome variable was child nutrition. Predictor variables represented two domains of father's involvement. Multivariable general linear modeling and multivariable logistic regression modeling were performed with the use of a combination of stepwise and hierarchical approaches in data analysis. RESULTS: The overall prevalence of underweight among children was 19.1%, and the prevalence of stunting was 14.4%. Children whose fathers did not bring them to a medical facility for immunization were about 1.7 times more likely to be underweight and stunted than those whose fathers did bring them for immunization after child's age, household economic status, and mother's education were controlled for. Father's involvement in housework was not found to be related to the prevalence of malnutrition. CONCLUSIONS: Paternal involvement in child immunization should be encouraged by health-care providers who manage immunization programs. PMID- 18510207 TI - Impact of pilot project of Rural Maintenance Programme (RMP) on destitute women: CARE, Bangladesh. AB - BACKGROUND: The rate of malnutrition among women in Bangladesh is high, but historically there has not been a specific program focusing on the improvement of the nutritional status of Bangladeshi women. OBJECTIVE: To observe changes in the nutritional status of destitute women of the Rural Maintenance Programme (RMP) by incorporating a health and nutrition intervention package with RMP ongoing activities. METHODS: An intervention study involving 1,275 poor destitute women was conducted from July 2004 to June 2005 in 17 districts in Bangladesh under two field offices, Mymensingh and Jessore, covering 8 and 9 districts, respectively. The respondents were divided into intervention, comparison, and control groups. All participants in the intervention and comparison groups were paid as part of the RMP and received weekly 30-minute nutrition interventions for 7 weeks in addition to routine training. The comparison group also received RMP training. The control group consisted of women with similar demographic characteristics to the intervention and comparison groups who did not receive pay or any intervention. The intervention was a unique combination of the three components of the UNICEF triangle model (food security, caring practices, and disease control). Data on socioeconomic and anthropometric characteristics, immunization, and vitamin A capsule intake were also collected with the use of a structured questionnaire. RESULTS: After the intervention, the mean body weight had significantly increased by 1,333 g in the intervention group and had decreased by 277 g in the control group and 147 g in the comparison group. The body mass index of women in the intervention group had also significantly increased at the end of the study (p < .001). There was a significant increase in the intake of iodized salt in the intervention group as well as increased immunization coverage in all groups. Intake of the first vitamin A capsule by children increased (from 60% to 97%) in the intervention group only. CONCLUSIONS: The nutrition pilot intervention was highly effective in improving the nutritional status of women in the RMP. PMID- 18510208 TI - [Role of endoscopic endonasal approach in surgery for skull base neoplasms]. PMID- 18510209 TI - [Prevention and management of complications of endoscopic surgery for nasal-skull base neoplasms]. AB - OBJECTIVE: To analyze the surgical complications of endoscopic nasal-skull base surgery. The secondary objective was to propose the preliminary strategies for prevention and treatment of complications. METHODS: One hundred and thirty two patients with nasal-skull base tumors undergoing endoscopic or endoscope-assisted surgery were included in this study. Surgical approaches included endoscopic endonasal transethmoidal approaches, endoscopic endonasal transseptal transsphenoidal approach, extended endoscopic endonasal transseptal transsphenoidal approach, endoscopic transmaxillary posttrial wall approach, extended endoscopic transmaxillary posttrial wall approach, endoscopic nasal lateral wall dissection, maxillary osteotomy approach and endoscopic transoropharyngeal approach. These approaches were selectively used to resect the tumors in the area of nasal-skull base. RESULTS: The total resection of the tumors was obtained in 104 patients (104/132, 78.8%), with 29.5% (39/132) incidence of complications, including profuse bleeding, nerve injury, cerebrospinal fluid leakage, diabetes insipidus, electrolyte imbalance, hyperglycemia, and psychological disturbance. No catastrophic complications, sequelae and operative mortality encountered. Four months to 8 years' follow up (median 3.0 years) indicated that recurrence rate of the benign tumor was 9% (9/100) without died case, and 3-year and 5-year survival rates of the malignant tumor were 75.0% and 55.6%, respectively. CONCLUSIONS: Strategies proved to be effective in reduction of the overall incidence of the complications, especially in minimizing the catastrophic complications and sequelae. The strategies were as follows: first, according to original site, extension and characteristics of the tumor, designing appropriate endoscopic approaches for the treatment of skull base tumor; second, recognizing reliable surgical access points and safe plane of the dissection; third, predicting surgical risks preoperatively and proposing the corresponding plan to avoid these risks; fourth, acquainted with the endoscopic skills and familiarized the skull base structures; lastly, ensuring the correct management of the interdisciplinary problems with close collaboration with the interdisciplinary medical personnels. PMID- 18510210 TI - [Investigation of surgical managements of olfactory neuroblastoma]. AB - OBJECTIVE: To evaluate the safety and efficacy of transnasal endoscopic resection and craniofacial resection through an external approach for olfactory neuroblastoma (ONB). METHODS: Thirty two patients with ONB treated between 1987 and 2006 were retrospectively reviewed. RESULTS: The patients were followed up for 8-135 months, the median follow-up time was 20 months. The longest follow-up time of patients treated by endoscope was 79 months, and patients treated by combined endoscope and transcranial surgery was 87 months. At Kadish stage B the 3-year survival rate of patients with transnasal endoscopic resection was 78.8% and at Kadish stage C it was 50.0%. At Kadish stage B the 3-year survival rate of patients with craniofacial resection through an external approach was 60.0% and at Kadish stage C it was 44.4%. The bleeding amounts in above two approaches were 140 ml and 450 ml. The average length of stay in hospital in transnasal endoscopic resection approach was markedly reduced (P < 0. 01). CONCLUSIONS: Olfactory neuroblastoma can be safely and effectively excised and reconstructed endoscopically with comparable degrees of tissue removal as with external approaches. The time of stay in hospital can be reduced and the surgical trauma can be diminished. PMID- 18510211 TI - [Treatment of cerebrospinal fluid rhinorrhea via transnasal endoscopic approach and its combination with frontal approach]. AB - OBJECTIVE: To summarize the clinical experience in diagnosing and managing cerebrospinal fluid (CSF) rhinorrhea via transnasal endoscopic approach and its combination with frontal approach. METHODS: A retrospective study of 58 patients with CSF rhinorrhea was conducted. Fifty eight cases were all under CT. Fifty six cases underwent surgical treatment Among them, 45 patients were treated with transnasal endoscopic approach, 11 whose cerebrospinal fistulas located in back wall of frontal sinus and orbital-frontal part with fronto-rhinal approach. Two were without any surgical treatment. Among 56 cases who underwent surgical treatment, 31 cases were under normal CT, of which 25 were accurate. Another 25 cases were under thin-section spiral CT scan and three-dimensional reconstruction, of which 23 were accurate. Eight cases locating the fistulas inaccurately by CT found the fistulas by operation. RESULTS: Postoperative follow up lasted from 6 months to 5 years, a median follow-up period of 3 years. Among 45 cases with transnasal endoscopic approach, 43 were cured after the first attempt, one was cured after the second attempt; one died because of the intracranial infection. Among 11 cases with fronto-rhinal reossification, 10 were cured after the first attempt, one with orbital-frontal absence after the fifth attempt. Two left hospital and lost following-up without any surgical treatment. CONCLUSIONS: Thin-section spiral CT scan and three-dimensional reconstruction make the leak locating more accurate. Combination of frontal approach may deal with transnasal endoscopic surgery's demerit to the unreachable site and enhance the achievement ratio of the first attempt. PMID- 18510212 TI - [Endoscopic endonasal surgery for meningoencephalocele]. AB - OBJECTIVE: To investigate the management experience with transnasal endoscopic technique for meningoencephalocele. METHODS: Nine patients with endonasal encephalomeningocele were managed by transnasal endoscopic surgery, and the skull base defect was repaired by fascia. RESULTS: Eight cases were successfully managed at the time of the first operation, and no relapse case was found during 1 to 4 years follow-up. Only one case of a two years old child relapsed with cerebrospinal fluid rhinorrhea one month after operation. During the second operation, titanium mesh uncovering was found, and replacement of titanium mesh by fascia via skull base defect was done, without relapse one and half years after the second operation. Another case of a one year old child got a fever one day after operation, but no white blood cell was found in the cerebrospinal fluid, and the temperature recovered to normal after release cerebrospinal fluid management. There were no complications of cranial infection, hemorrhage, edema and water retention in brain to be found in all cases. CONCLUSIONS: It is not only minimally invasive, safety and efficiency of transnasal endoscopic technique for meningoencephalocele, but also had a clear operating view for better recolonization of the position of leak and the structure of operating field, therefore, transnasal endoscopic technique is the first choice for the management of endonasal encephalomeningocele. The accurate localization of leak and selection of the appropriate repairing materials are the key point for the successful operation. PMID- 18510213 TI - [Outcome of cochlear implantation in prelingual pediatric auditory neuropathy]. AB - OBJECTIVE: To explore the electrophysiological results and rehabilitation outcome of two prelingually deafened pediatric cochlear implant patients with auditory neuropathy. METHODS: Preoperative audiological evaluation, intra-postoperative electrically evoked auditory brainstem response (EABR) and neural response telemetry (NRT) record for the two cases were conducted in Beijing Tongren Hospital. A one year follow-up was performed. Data collected before and at 6,12 month intervals after implantation were compared with that from control pediatric cochlear implant patients matched for the same duration of implant use as this two cases. RESULTS: The two children implanted had not had any postoperative medical or cochlear implant device complications. Intraoperative EABR and NRT were elicited in case 1 with unrepeatable waveforms. After 12 months of training, Case 1 had shown significant improvements in sound detection, speech perception abilities and communication skills, which was better than the control group, and the electrophysiological results became normal. Case 2 had also benefited from cochlear implantation, even though no recognizable NRT was found until he returned 12 month after the operation. CONTUSIONS: The desynchronization of auditory path had been changed after the electrical stimulation ongoing 12 months for children with auditory neuropathy. The two children had not had any complications postoperatively, and each child had shown improved listening and communication skills. Cochlear implantation could help patients with auditory neuropathy to improve their communication skill and go back to the main stream. PMID- 18510214 TI - [Comparative study of pure tone stimuli and Chinese speech stimuli on auditory event-related potentials]. AB - OBJECTIVE: To compare the characteristics of auditory event-related potentials (AERP) evoked by pure tone stimuli and Chinese speech stimuli respectively, and to explore the feasibility of using Chinese speech stimuli to evoke AERP for Chinese. METHODS: AERP were tested by both Chinese speech and pure tone as stimuli in normal young participants (83 ears in 44 young postgraduate students), then each AERP wave form were scored. The latencies, amplitudes and scores of AERP evoked by speech stimuli were compared with by pure tone stimuli. RESULTS: Typical waves of AERP were recorded and identified more easily with speech stimuli than pure tone stimuli, moreover, the differences were statistically significance (X2 = 4.0, P = 0.039). The latency and amplitude of P3 evoked by both Chinese speech stimuli and pure tone stimuli in the 72 ears were no significant difference (P > 0.05). But the mean scores of AERP evoked by speech stimuli were significantly higher than those evoked by pure tone stimuli (t = 6.57, P = 0.000). N2 and P3 latency in left ear evoked by speech stimuli were significantly shorter than those evoked by pure tone stimuli (P = 0.002, P = 0.003). However there were no significant differences in right ear(P > 0.05). CONCLUSIONS: Chinese speech stimuli were more consistent with requests of AERP test and custom of Chinese spoken language, so the Chinese speech stimuli was more available for Chinese's AERP test than pure tone stimuli. PMID- 18510215 TI - [Ophthalmic complication and management of endoscopic sinus surgery]. AB - OBJECTIVE: To investigate the possible reasons and the treating experiences of ophthalmic complications in order to elevate the attention of the nasal endoscopic surgeon. METHODS: This study was involved 8 categories in 22 cases which had typical characteristic of ophthalmic complications of endoscopic sinus surgery, including injury of lamina papyracea, obstructive cyst of frontal and ethmoid sinuses, orbital infection, injuries of lacrimal passages, injury of extraocular muscles, orbital hemorrhage, optic nerve injury and arterial embolism of optic fundi. The patients were given corresponding treatment. RESULTS: Injury of lamina papyracea was cured by medical treatment while orbital infection, injury of lacrimal passages, obstructive cyst of frontal and ethmoid sinuses were completely recovered by endoscopic surgery. In one of the cases, the injured optic nerve had recovered after transnasal optic and orbital apex decompression but the other 6 sides of 6 patients had not been improved. In one case who had suffered from injury of extraocular muscles, their diplopia disappeared mostly, but the other cases had a slight diplopia after surgery. One case who had visual descending caused by orbital hemorrhage recovered. Two cases of visual loss caused by intraorbital hemorrhage and arterial embolism of optic fundi respectively were not improved. CONCLUSIONS: It is very difficult to cure if the patients lose the sight because of optic nerve injury, orbital hemorrhage and the ischemia of orbit, however, if the patients had remnant vision the prognosis should be much better. PMID- 18510216 TI - [Case-control survey on risk factors of benign vocal fold lesions]. AB - OBJECTIVE: To investigate the risk factors that may relate with benign vocal fold lesions including vocal fold nodule, vocal fold polyp, chronic laryngitis and Reinke's edema METHODS: In present series, 321 cases who were performed laryngoscope were invited to participate the survey. Among them 168 cases with benign vocal fold lesions composed the case group. Another 153 cases with normal larynx composed the control group. Each case were undertook the same questionnaire. Logistic regression analysis was preformed to investigate the possible risk factors. RESULTS: The result demonstrated the occurring of benign vocal fold lesions positively correlated to five factors, including occupation, work or residence environment noise, alcohol-consuming, voice-using hours per day and abuse of voice. Occupations with intensive voice-use were more vulnerable to developing these disorders. Occurring risk of occupations type II with moderate voice-use was 1.934 times than that of occupations type I with lesser voice-use (OR = 1.934). And risk of occupations type III with upper voice-use was 2.633 times than that of type I. Risk raised 1.302 times with each more hour of voice use per day. OR of the following factors of voice abuse, environment noise, alcohol-consuming was 4.744, 2.115 and 2.177, respectively. CONCLUSIONS: The result suggested that people should abstain from alcohol, lowering the environment noise, prevent overuse and abuse of voice in order to decrease the prevalence of these disorders, which is especially important for the professional voice users, e. g. teachers or managers. The essential therapy for these disorders is to correct bad phonation habits. PMID- 18510217 TI - [Relevance between clinical behavior and bionomical findings of acoustic neuromas]. AB - OBJECTIVE: To evaluate the relationship between labeling index (LI) Ki-67, proliferating cell nuclear antigen (PCNA) and transforming growth factor-beta1 (TGF-beta1) with the clinical behavior of acoustic neuroma. METHODS: Expression of Ki-67, PCNA and TGF-beta1 was detected by immunohistochemistry in 53 specimens of acoustic neuromas. The relationship among tumor proliferation, histological representation, size of tumor, clinical proliferation index of tumor and tumor proliferation activity were analyzed. RESULTS: In all 53 cases, the positive rate of Ki-67 was 77.4% (41/53) but the positive rate of PCNA was 84.9% (45/53). There was significant difference between the proliferate index, clinic growth rate and course of disease (t = 2.14, t = 2.70; P < 0.05). The positive rate of TGF-beta1 was 83.0% (44/53). The correlation of TGF-beta1 with LI (Ki-67) was significant difference (r = 0.36, P < 0.05). Cystic degeneration often occurred in large-size tumor (Z = 4.44, P < 0.05). There was no significant relationship between the expression of LI (Ki-67), LI (PCNA) and TGF-beta1 and the course of disease as well as between the cystic degeneration and the non-cystic degeneration. Although clinic growth rate of cystic degeneration was bigger than that of non-cystic degeneration, there was not statistically significant. CONCLUSIONS: Ki-67 and PCNA are reflected proliferation activities of tumor cells in acoustic neuromas. Cell proliferation-labeling index LI (PCNA) was related with clinical growth rates. TGF-beta1 might participate in the biological behavior of acoustic neuroma. Cystic degeneration was one of special pattern of acoustic neuroma, however, tumor enlargement might due to the volume of the cystic but unrelated to fast proliferation of parenchyma cell. PMID- 18510218 TI - [Role of aquaporin 1 in the migration of eosinophils from asthmatic guinea pigs]. AB - OBJECTIVE: To investigate the role of aquaporin 1 (AQP1) in the migration of eosinophils (EOS) and to determine if AQP-1 can be viewed as the chemotactic activity marker of EOS. METHODS: Asthma model of guinea pigs were developed and EOS were purified from both peripheral blood and bronchoalveolar lavage fluid (BALF). The smears of EOS were studied by in situ hybridization for determining AQP1 mRNA and immunofluorescence under laser scanning confocal microscope for determining AQP1 protein. RESULTS: AQP1 was found expressed in EOS both from peripheral blood and from BALF. Compared with the expression of AQP1 mRNA (mean grey value 109.200 +/- 5.756, x +/- s) and protein (average fluorescence intensity 279.926 +/- 11.293) in EOS from BALF, there was stronger expression of AQP1 mRNA (92.904 +/- 3.290) and protein (425.081 +/- 17.474) in EOS from peripheral blood. The difference both of AQP1 mRNA (t = 9.519, P < 0.05) and protein(t = 27.020, P < 0.05) were considered statistically significant respectively. CONCLUSIONS: AQP1 plays a crucial role in EOS movement. It is possible that EOS produce more AQP1 protein to accelerate its migration to inflammatory tissues under allergic disease and EOS with AQP1 highly expressed are activated. AQP1 can be viewed as the chemotactic activity marker of EOS. PMID- 18510219 TI - [Treatment of allergic rhinitis rats by intranasal interferon gamma]. AB - OBJECTIVE: To investigate the effects and mechanism of intranasal interferon gamma (IFN-gamma) in the treatment of allergic rhinitis. METHODS: Ovalbumin (OVA) absorbed to aluminum hydroxide was used to construct the allergic rhinitis model (group C), and the normal control group (group A), the allergic rhinitis model group (group B) and beclomethasone dipropionate group (group D) consisted of 8 rats for each. PBS 50 microl was absorbed to group B, IFN-gamma 1 microg was absorbed to group C and beclomethasone dipropionate 3.5 microg was absorbed to group D on day 31 to day 38 once daily once nasal cavity. The nasal lavage fluid was collected on day 39, and the cellular constituents, levels of interleukin-4 (IL-4), interleukin-5 (IL-5) and IgE were determined, together with the pathologic changes and expression of GATA-3 were observed. RESULTS: Decrease of eosinophils [(0.005 +/- 0.003) x 10(4)/ml, x +/- s] was seen in nasal lavage fluid of group C as comparing with group B [(0.225 +/- 0.060) x 10(4)/ml, (P < 0.01)], and the levels of IL-4 (7.8 +/- 3.5) pg/ml and IL-5 (12.5 +/- 4.3) pg/ml decreased significantly in comparing with group B (P < 0.01). The serum levels of total IgE (38.5 +/- 9.6) microg/ml and ovalbumin-specific IgE (19.8 +/- 5.4) IU/ml decreased significantly in comparing with those of group B (P < 0.01). In group B, mucosal congestion and edema thickening with inflammatory cells infiltration mainly of eosinophils; in group C, the above mentioned changes were much more ameliorated. Immunohistochemistry showed significant increase of GATA-3 expression in the nasal tissue of group B but much lesser than that in group C. CONCLUSIONS: IFN-gamma can inhibit the composition of IL-4 and IL-5, and inhibit the airway inflammation with eosinophilic infiltration and the serum levels of total IgE and ovalbumin specific IgE, probably through the mechanism of restraining the Th2 reaction by blockade of GATA-3 expression in the nasal tissue. PMID- 18510220 TI - [Clinical evaluation of over-under myringoplasty technique in large tympanic membrane perforation]. PMID- 18510221 TI - [Investigation of anatomical characteristics of hypoglossal nerve and lingual artery to in tongue base]. PMID- 18510222 TI - [Application of ultrasonic scalpel in tonsillectomy]. PMID- 18510223 TI - [Analysis of misdiagnosis of two patients in myasthenia gravis]. PMID- 18510224 TI - [One case of ten-years bronchial foreign body]. PMID- 18510225 TI - [Primary laryngeal small cell carcinoma: report of 5 cases]. PMID- 18510226 TI - [Malignant myoepithelioma of the left maxillary sinus]. PMID- 18510227 TI - [Diagnosis and treatment of primary parathyroid carcinoma: report of 2 cases]. PMID- 18510228 TI - [One case of eosinophilic granuloma]. PMID- 18510229 TI - [The necessity of neck dissection determined by negative post chemotherapy/radiotherapy FDG-PET in patients with head and neck cancer]. PMID- 18510231 TI - [Fully to bear the social responsibility of the academic journal]. PMID- 18510230 TI - [Diagnosis and treatment of carotid body tumor]. PMID- 18510232 TI - [Marching forward from the way beginning]. PMID- 18510233 TI - [The strategy for the prevention of diabetic blindness]. AB - Diabetic retinopathy (DR) is one of the most important causes of blindness in China as well as in the world. Recent years, the screening of DR and the therapy based on the evidence-based medicine are emphasized for effectively controlling the increase of the prevalence of DR. Extensive screening of DR in the community and secondary hospitals, periodic follow-up and evaluation of the patients are the key measures for reducing the rate of the visual impairment. The new international classification of DR is helpful for improving the communication among primary eye care workers, endocrine physicians, and ophthalmologists. The outcomes of several multi-center prospective randomized clinical trials on the photocoagulation, vitrectomy and systematic therapy for DR are useful for the early prevention and management of DR. If the therapy is based on the evidence based medicine, the visual impairment caused by DR may be controlled effectively. PMID- 18510234 TI - [Emphasizing the early prevention and treatment for diabetic retinopathy]. AB - Diabetic retinopathy is a complication of diabetes which is difficult to reverse and can result in blindness. Ophthalmologists in China are facing quite a challenge according to the weak prevention and treatment system of DR. At present, the emphasis should be placed on the early prevention and treatment, but the appropriate guidelines founded on evidence-based medicine still have been absent. Currently, we should deeply realize the problems existing in early prevention and treatment of DR in China, fully recognize the necessity and importance of it, actively spread the new international classification, adopt the ways of public health, start by primary health care and fully cooperate with different fields of the society to strengthen the practice and research work on the early prevention and treatment of DR. PMID- 18510235 TI - [Comparison of fundus photography and fluorescein angiography in grading diabetic retinopathy]. AB - OBJECTIVE: To study the consistency of the color fundus photography and fluorescein angiography for grading the diabetic retinopathy. METHODS: It was a retrospective case series. The agreement study was conducted in a series of diabetic patients, who were excluded from the mild non-proliferative diabetic retinopathy (NPDR) and the severe proliferative diabetic retinopathy (PDR). Ninety-nine cases (188 eyes) met the selected criteria Five 50 degrees non stereoscopic photography for each eye were taken in all cases before FFA was performed. The diabetic retinopathy was graded from the fundus photography and FFA images according to international diabetic retinopathy disease severity scale. The kappa value was calculated according to the grading outcomes from the fundus photography and FFA images to estimate the agreement of the grading outcomes. RESULTS: The fundus photography showed 59 moderate NPDR (31.4%), 76 severe NPDR (40.4%) and 53 PDR eyes (28.2%) respectively, FFA showed 50 moderate NPDR (26.6%), 72 severe NPDR (38.3%) and 66 PDR (35.1%) respectively. The agreement for the grading of DR by the fundus photography and FFA was substantial (K = 0.601). In the evaluation whether it was necessary to implement PRP (more serious than NPDR and PDR), there was higher agreement between them (K = 0.652). CONCLUSIONS: Five 50 degrees photography can provides the support of the application of PRP for diabetic patients, FFA may be earlier and relatively accuracy in the diagnose of diabetic retinopathy. PMID- 18510236 TI - [Four-port pars plana vitrectomy for severe proliferative diabetic retinopathy]. AB - OBJECTIVE: To investigate the 4-port pars plana vitrectomy in eyes with severe proliferative diabetic retinopathy (PDR). METHODS: It was a case-control study. Twenty-eight eyes in 27 patients with extensive fibrovascular proliferation associated with PDR were retrospectively collected, who were undergone 4-port pars plana vitreous surgery with bimanual manipulation techniques, such as membrane dissections and enbloc membranectomy. The control group consisted of 30 eyes in 30 patients with PDR which were undergone 3-port pars plana vitrectomy by the same surgeon. RESULTS: Twenty-eight eyes were undergone membrane dissection and enbloc membranectomy smoothly during 4-port pars plana vitrectomy, 2 iatrogenic holes occurred in 1 eye. During the follow up 7 months to 4.5 years, the retina was fully attached in all eyes, visual acuity had improved except 1 eye which complicated with neovascular glaucoma. In the control group, membranes partially remained in 2 eyes, 4 iatrogenic holes appeared in 3 eyes, neovascular glaucoma occurred in 3 eyes, the retina was reattached during the follow-up time from 12 to 34 months. CONCLUSIONS: For severe proliferative diabetic retinopathy, the 4-port pars plana vitrectomy with bimanual manipulations of membrane peeling is safe and efficiency. PMID- 18510237 TI - [Clinical features and surgical outcomes of children with rhegmatogenous retinal detachments]. AB - OBJECTIVE: To describe the clinical features and surgical and visual outcomes in a series of children with rhegmatogenous retinal detachments (RRD). METHODS: Retrospective case series of pediatric patients (14 years old or younger) with RRD who underwent primary surgery at a tertiary referral center between May 1995 and May 2006. Patients with perforating ocular trauma were excluded. RESULTS: Thirty-six eyes of 33 patients were included in this study. The median age was 12 years, and 26 patients (78.8%) were male. Bilateral RRD was present in 10 patients (30.3%); 22 patients (66.7%) had some form of bilateral ocular pathology. At least one predisposing factor could be identified in 31 (86.1%) of eyes examined. Thirteen eyes (36.1%) experienced trauma, 11 eyes (30.6%) had structural abnormalities, high myopia and eyes underwent previous intraocular surgery were 7 (19.4%) each, and 3 eyes (8.3%) had inflammatory fundus disorders. Macular detachment was found in 28 eyes (77.8%). Median follow-up was 12 months. At the end of follow-up for this report, silicone oil was still in place in 4 eyes. Retinal reattachment was ultimately achieved in 29 of the 32 eyes (90.6%) with a mean of 1.9 surgeries per eye, however, visual recovery was modest. Predictors of a poor visual outcome were: hand motions or worse in preoperative vision (P = 0.001), macular-off retinal detachment (P = 0.003), proliferative vitreoretinopathy grade C or worse (P = 0.000), the need for vitrectomy surgery (P = 0.002), and the use of silicone oil (P = 0.005). CONCLUSIONS: Trauma and congenital or developmental anomalies were the most common predisposing factors for pediatric RRD in this series. Retinal reattachment can be achieved in most cases with modern vitreoretinal surgical techniques. Predictors of visual outcomes are similar to those observed in adults. Regular follow-up for children at risk of developing RRD is necessary for early detection. PMID- 18510238 TI - [Vitrectomy in vitreo-retinal complications associated with intermediate uveitis]. AB - OBJECTIVE: To investigate the effects of vitrectomy in vitreo-retinal complications associated with intermediate uveitis. METHODS: Retrospective case series of sixteen eyes of 16 patients in vitreo-retinal disease associated with intermediate uveitis in a 3-year period from Mar 2002 to Jun 2005 were included in the study. They were treated with vitrectomy and the mean follow-up was (14.25 +/- 7.90) months (range: 5-32 months). Visual acuity in final follow-up, post operative complications and the recurrence of intermediate uveitis were retrospectively reviewed. RESULTS: Four patients were associated with tuberculosis, rheumatoid arthritis, multiple sclerosis or Behcet disease, respectively. The remaining 12 cases had idiopathic diseases. Pre-operatively, all patients were treated with steroids for a long time and the mean treatment time was (9.94 +/- 2.67) months (range: 6-16 months). Pre-operative vitreo retinal complications included severe vitreous organization (5 eyes), tractional retinal detachment (6 eyes), rhegmatogenous retinal detachment (1 eye), vitreous hemorrhages (2 eyes), epimacular membrane (2 eyes) accompanied with vitreous tissue, and peripheral retinal neovascularization (16 eyes). In the post operative period, tractional retinal detachment in one eye and complicated cataract in 3 eyes were observed. Post-operatively, only 4 cases need long-term immunosuppression therapy (more than 6 months). Fourteen of 16 eyes achieved a final visual acuity equal to or better than baseline (X2 = 4.923, P < 0.05). Recurrent intermediate uveitis was not found in these patients. CONCLUSIONS: The results of this study suggest that pars plana vitrectomy may have a beneficial effect on the intermediate uveitis which was severe or uncontrolled by immunosuppressive drugs and accompanied with vitreo-retinal complications. The beneficial effects include improving visual acuity, reducing need for long-term immunosuppression treatment. PMID- 18510239 TI - [To observe clinical effect of accommodative IOL on different age patients]. AB - OBJECTIVE: To observe clinical effect of accommodative IOL on different age patients. METHODS: Retrospective case series of fifty-four patients implanted with accommodative IOL were divided into three groups according to their age: 39 60, 60-75 and older than 75 years. Three months after the cataract surgery, distant visual acuity, near visual acuity and defocusing with minus glasses (D) were examined and depth of anterior chamber was measured by ultrasound biomicroscope. RESULT: There were no statistically significant differences in distant visual acuity among these three groups. Near visual acuity in older than 75 group was significantly lower than the other two groups. There were no statistically significant differences in the depth of anterior chamber among these three groups. When patients watched the target moving from 5 meters away to near, the change of depth of anterior chamber in older than 75 group was significantly less than that in the other two groups. Accommodation power in older than 75 group was significantly lower than that in the other two groups as measured by defocusing with minus glass (D). CONCLUSION: Patients implanted with accommodative IOL can get good distant and near visual acuities. Younger patients may obtain more benefit from accommodative IOL such as obtaining higher accommodative power. PMID- 18510240 TI - [Measurement of visual acuity at different distances after implantation of ReSTOR multifocal intraocular lenses]. AB - OBJECTIVE: To measure visual acuity (VA) at different distances after implantation of ReSTOR multifocal intraocular lens (MIOL) and to evaluate the variation of VA with different distances. METHODS: It was a prospective case series study. Thirty eyes of 22 patients received the ReSTOR MIOL in this prospective case series study. VA was measured at several distances, and was recorded in a defocused range of 0 to -4.5 diopters (D) with gradually reduced minus spectacles based on distant refraction. A questionnaire was used to assess the satisfaction about VA of different distances. The mean follow-up time was three months. RESULTS: Postoperatively, the manifest refraction spherical equivalent was within +/- 1.00 D in 28 of 30 eyes (93.3%) and the astigmatism was within 1.00 D in 23 of 30 eyes (76.7%). The mean uncorrected distance (5 m), intermediate (60 cm) and near (33 cm) VA were 0.91, 0.40 and 0.64, respectively. The mean VA at 0 D, -1.50 D and -3.00 D pseudoaccommodation were 1.00, 0.49 and 0.79, respectively. On the questionnaire, five patients complained of unreality when walking, and two computer workers complained that they could not see screen clearly, and one hair-cutter complained that he could not see hair clearly. Self score of satisfaction about distant, intermediate and near VA were 8.0, 7.5 and 8.0, respectively. CONCLUSIONS: Implantation of the ReSTOR MIOL offers excellent distant and near VA. The VA decreases significantly at intermediate range as compared to distant and near vision, which might reduce the quality of life for patients who need excellent vision at intermediate range. PMID- 18510241 TI - [Clinical and pathological analysis of 2639 cases of eyelid tumors]. AB - OBJECTIVE: To describe the histopathologic spectrum and patient characteristics of eyelid tumors in Beijing Tongren Hospital. METHODS: It was a claims database analysis. A total of 2639 eyelid tumors verified histopathologically from January 1997 to December 2006 were included in the study. RESULTS: Of the 2639 eyelid tumors, the 5 most common eyelid benign lesions were inflammatory lesion, melanocytic nevus, papilloma, dermoid cyst and epidermoid cyst, and epithelial cyst. The 5 leading eyelid malignant tumors were basal cell carcinoma, sebaceous gland carcinoma, lymphoma, squamous cell carcinoma and melanoma. The mean age at diagnosis was 61 years for basal cell carcinoma and sebaceous gland carcinoma, 57 years and 52 years for squamous cell carcinoma and melanoma, respectively, and 48 years for lymphoma. There was no significant sex predilection in basal cell carcinoma and sebaceous gland carcinoma. Melanoma and lymphoma occurred more commonly in women, whereas squamous cell carcinoma occurred more commonly in men. CONCLUSIONS: Basal cell carcinoma and sebaceous gland carcinoma were the most common malignant eyelid tumors, and lymphoma ranked third and had an increasing trend. The malignant tumors occurred predominantly in the elderly of 60 years and above. PMID- 18510242 TI - [Clinicopathologic observation of orbital natural killer-T cell lymphoma]. AB - OBJECTIVE: To investigate clinical manifestation, diagnosis and treatment of orbital natural killer (NK)-T cell lymphoma. METHODS: It was a retrospective case series. Seven orbital NK-T cell lymphoma patients confirmed by surgical biopsies were collected during the past 22 years. We reviewed the records, surgical and treatment procedures. Surgical specimens were studied with HE staining, immunohistochemical staining and molecular biological analysis. RESULTS: These patients had proptosis, eye motive inhibition or fixation and visual acuity was decreased or even without light perception. Skin of inner canthus and eyelids appeared red and swollen, with ulceration and cavity formation. CT scan revealed that the tumor showed uneven density and an unclear border. Tremendous lymphocyte infiltration and tissue necrosis in the tumor were observed in the biopsy tissue. LCA, CD45RO and CD57 immunohistochemical staining revealed positive results. Clonal T-cell-receptor gene rearrangements of two patients showed negative results and the Epstein-Barr virus was detected. CONCLUSIONS: Orbital NK-T cell lymphoma is a rare disease. The characteristics of this disease include a highly aggressive clinical course, severe destruction and a poor prognosis. The final diagnosis depends on HE staining, immunohistochemical staining and molecular biological examination. PMID- 18510243 TI - [Clinical features of aniridia]. AB - OBJECTIVE: To explore the various clinical features of aniridia and its treatment. METHODS: It was a retrospective case series. All the 8 cases of 16 aniridia patients in our department from January, 1984 to January, 2007 were assayed. All the patients were bilateral aniridia suffering from abnormal vision and manifested with various degree of cataract, 5 eyes manifested with refractory glaucoma, 8 eyes with horizontal nystagmus. These patients underwent cataract extraction and intraocular lens implantation, trabeculectomy, ciliary process laser photocoagulation, in some of patients glass and cover strabismus exercise, color contact lens therapy were applied. RESULTS: Vision acuity in all of patients with severe cataract were improved after cataract surgery. One glaucoma eye underwent trabeculectomy, but failed to control the IOP and then received ciliary process laser photocoagulation. Three glaucoma eyes only underwent ciliary process laser photocoagulation, 1 eye received topical anti-glaucoma eye drops. The IOP of 3 eyes was controlled. However, the IOP of 2 eyes was not to be controlled. Vision acuity was improved in 4 strabismus eyes after wearing glass and strabismus exercise, the symptom of photophobia was disappeared in 2 eyes after wearing a pair of color contact lens. CONCLUSION: Although it is hard selection to select the proper treatment for aniridia, most of the cases can obtain a better vision, and relief of the symptom after proper treatment. PMID- 18510244 TI - [The research of the protection of recombinant human erythropoietin in human RPE cells by light-induced injuries]. AB - OBJECTIVE: The aim of this study was to assess the protection of recombinant human erythropoietin (rhEPO) in light-induced injures in human retinal pigment epithelial (RPE) cells. METHODS: It was a experimental study. Cultured human RPE cells were exposed to light of 8w 2000 +/- 500 Lux for 12 hours. The 3-(4,5 dimethylthiazole-2y1)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay were used to assess the effects of rhEPO in light-induced injury on human RPE cells. The effect of inhibiting apoptosis of rhEPO was detected by AnnexinV fluorescein isothiocyanate/Propidium iodium labeling and flow cytometry. The enzyme linked immunosorbant assay (ELISA) and immunocytochemical staining were used to assess the expressions of caspase-3 treated by different doses of rhEPO in light-induced injury on human RPE cells and examine the protective mechanism of rhEPO by treatment with AG490 (the special inhibitor of jak2). RESULTS: There was a significant increase of inhibiting apoptosis in every rhEPO group, and cell viability was the highest in 40 U/ml rhEPO group, the value was 4.93 +/- 1.45/ml. The decrease in expression of caspase-3 was the most obvious in 40 U/ml rhEPO group, in which the value was 0.125 +/- 0.029 ng/ml. There was a significant increased effect on inhibiting apoptosis in every rhEPO group, and it was the most conspicuous in 40 U/ml rhEPO group. But these increased cell viability and effect on inhibiting apoptosis in rhEPO group were restrained by AG490, in which the value of apoptosis was 11.29 +/- 2.11/ml and the density of caspase-3 increased to 0.362 +/- 0.042 ng/ml. CONCLUSIONS: It is suggested that rhEPO can protect human RPE cells from the light-induced injures. Its protective mechanism is principally mediated by the EPO-EPOR pathway, which subsequently leads to jak2 activation. PMID- 18510245 TI - [CTLA4-FasL protein for the prevention of immune rejection in mouse corneal transplantation]. AB - OBJECTIVE: To investigate the therapeutic effect of double-functioned CTLA4-FasL protein for the prevention of immune rejection in murine corneal allografts. METHODS: It was a experimental study. C57BL/6 mice (n = 45) were as donors and BALB/c mice (n = 90) as recipients. BALB/c mice accepted penetrating keratoplasty were randomly divided into 3 groups including no therapy (A), CsA DDS implanted in anterior chamber (B), 10 microg/mL CTLA4-FasL (C). Survival time of corneal allografts was observed and routine assays were performed including immunohistochemistry, reverse-transcription polymerase chain reaction (RT-PCR), TdT-mediated dUTP nick end labeling (TUNEL). RESULTS: Survival time of corneal transplants was (14.3 +/- 1.3) days, (58.0 +/- 2.8) days, (106.3 +/- 17.5) days respectively. There was significant difference between groups in statistics analysis (P = 0.000). Inflammatory cells, predominant for CD4 + T cells, showed an increasing tendency for rejected corneal allografts in A group, peaked on postoperative 7 d and then decreased in C group compared with invisible in B group. CD80 or CD86 was detected on postoperative 3 d or 7 d respectively in excised cornea and iris of both A and B group, while either showed weakening expression in C group. On postoperative 14 d, IL-10, TNF-alpha, IFN-gamma mRNA were only detected in rejected cornea allograft of A group other than clear allografts of B or C group. Compared with no apoptosis in the other groups, abundant apoptosis cells were visible in cornea and iris of C group on postoperative 7 d. CONCLUSIONS: CTLA4-FasL is able to prolong survival time of corneal allografts by dual action mechanism to exert immunosuppressive effects, both blockade of CD28-CD80/86 pathway and induction of apoptosis of T cell. PMID- 18510246 TI - [The effects of the inhibitor of nuclear factor on lipopolysaccharide-induced cytokine expression in cultured human corneal fibroblasts]. AB - OBJECTIVE: To investigate the effects of pyrrolidine dithiocarbamate (PDTC) on Lipopolysaccharide (LPS)-mediated activation of nuclear factor kappa B (NF kappaB) and cytokine expression in cultured human corneal fibroblasts. METHODS: It was a experimental study. A completely random design was employed in this research. Human corneal fibroblasts (HCFs) were obtained from human specimen. HCFs were divided into three groups: control group (group 0 h), LPS alone group and PDTC treatment group. Cells were incubated with PDTC for 30 min in the PDTC pretreatment group before LPS challenged. At different time after LPS challenged, the activities of NF-kappaB were assessed by Western Blot analysis, the secretion of IL-6 and IL-8 from cultured corneal fibroblasts was measured with enzyme linked immunosorbent assays (ELISA); the mRNAs expression of IL-6 and IL-8 was determined by reverse transcription polymerase chain reaction (RT-PCR). The effects of PDTC on activation of NF-kappaB and the expression of IL-6 and IL-8 were also assessed in HCFs challenged with LPS. RESULTS: Compared with control group, NF-kappaB level was significantly enhanced in the nucleus in the LPS alone group, indicating that LPS mediated the activation of NF-kappaB in HCFs. The activation of NF-kappaB by LPS was markedly inhibited by PDTC (t1h = 9.3766, t2h = 15.9011, t4h = 12.5851, t8h = 10.8346, P < 0.01). Compared with control group, LPS increased IL-6 and IL-8 expression both mRNA and protein in HCFs. At the same time, PDTC partly inhibited the expression of IL-6 and IL-8 in corneal fibroblasts induced by LPS. These inhibitory effects were significant at both the mRNA and protein levels (protein of IL-6: t1h = 7.9154, t2h = 10.863, t4h = 8.2451, t8h = 13.5063. protein of IL-8: t1h = 8.5663, t2h = 20.5169, t4h = 25.1580, t8h = 34.8699. mRNA of IL-6: t1h = 12.0235, t2h = 13.2894, t4h = 24.0799, t8h = 27.2261. mRNA of IL-8: t1h = 20.9424, t2h = 24.1314, t4h = 29.8580, t8h = 47.9442. P < 0.01). CONCLUSION: These results suggest that LPS can activation of NF-kappaB in cultured HCFs and PDTC partly inhibits NF-kappaB activation. PMID- 18510247 TI - [Expression of mGluR1 at primary visual cortex of monocular deprivation amblyopia rat and the observing of ultrastructure]. AB - OBJECTIVE: To explore the regulation of expression of mGluR1 and the changes of neuron ultrastructure at primary visual cortex of monocular deprivation amblyopia rat within cortical period. METHODS: Taking randomized concurrent controlled trail. Establishing the model of monocular deprivation amblyopia rat. After proving the model successful by PVEP, all of the rats were randomly divided into three groups: normal visual cortex, experimental visual cortex and experimental opposite visual cortex. Immunocytochemical technology, electron microscope, photography microscope, computer image analysis, SPSS 11.5 and ANOV were used to get the results. RESULTS: Compared with the layer IV of normal visual cortex and experimental opposite visual cortex, the area of immunopositive neurons in layer IV of experimental visual cortex are deficiency, there is significant difference between them (P < 0.01). There are no significant differences between the other four corresponding layers (P > 0.05). Morphological abnormals were found in layer IV of experimental visual cortex by observing of ultrastructure. CONCLUSION: The expression of mGluR1 in layer IV of primary visual cortex of monocular deprivation amblyopia is reduced. There are morphological abnormals happened in layer IV of primary visual cortex of monocular derivation amblyopia. Reduced afference of nerve pulse because of monocular deprivation leads to the expression deficiency of mGluR1 in layer IV of the primary visual cortex, then synaptic plasticity happened, then neurons atrophy occurred may be one of the etiopathogenesis of amblyopia. PMID- 18510248 TI - [Progress of research on pathogenesis and medical treatment of diabetic retinopathy]. AB - Diabetic retinopathy is a common complication of diabetes mellitus. Studies on the pathogenesis of diabetic retinopathy are important for the prevention and treatment of this disease. Neurodegeneration and microvascular dysfunction are the major events of diabetic retinopathy happened in the early stage of diabetes mellitus. The pathologic changes include neuron apoptosis, release of inflammatory mediators and impaired glutamate metabolism. Diabetic retinopathy has many pathological changes with inflammatory characteristics. Because of the adhesion and migration of the leucocyte, increased permeability of blood vessel, haemodynamic change and many inflammatory mediators involved in the process of diabetic retinopathy, more and more researchers believe that diabetic retinopathy is a chronic inflammatory disease. These studies provide new notion of mechanisms of diabetic retinopathy and theoretical foundation for the treatment of this disease. Further studies on the pathogenesis of diabetic retinopathy can provide novel procedures for the prevention and treatment of this disease. PMID- 18510249 TI - [Intraocular lens power calculations following keratorefractive surgery]. AB - Intraocular lens power calculation after corneal refractive surgery is complicated. It is common to have a great postoperative deviation with the traditional method. The reasons for IOL power calculation error after corneal refractive surgery are analysed and many solutions are listed in this article, which we hope could provide basis for these patients. PMID- 18510250 TI - Crossing the threshold, finding a balance. PMID- 18510251 TI - Evaluating ADL measures from an occupational therapy perspective. AB - BACKGROUND: Measures reflecting occupational therapy's conceptualization of occupational performance support the profession's contribution to evidence-based practice and fiscal accountability. PURPOSE: This study compared measures of performance-based activities of daily living (ADL) with principles of occupational therapy practice and intended outcomes. METHODS: Using an action research study design, occupational therapists and researchers (N= 13) systematically clarified the clinical problem, identified occupational therapy principles inherent to the assessment of daily living activity via nominal group technique; defined the key principles as constructs; and reframed these constructs as a questionnaire against which 18 published standardized ADL measures were evaluated. FINDINGS: Participants identified six measures as most congruent with principles of occupational therapy practice: ADL Profile, Assessment of Motor and Process Skills, Functional Performance Measure, Rivermead ADL Assessment, Edmans ADL Index, and Melville-Nelson Self-Care Assessment. IMPLICATIONS: Findings guide occupational therapists' search and use of performance-based ADL measures that demonstrate the profession's distinct health care contribution. PMID- 18510252 TI - Falls and the physical environment: a review and a new multifactorial falls-risk conceptual framework. AB - BACKGROUND: Fall-related injuries result in significant physical and psychological suffering to the affected individuals. The physical environment is considered to have an important role in falls. PURPOSE: To conduct extensive review of and synthesize related literature, and to develop a conceptual framework to explain the relationship among falls, the physical environment, and older adults. METHODS: Review of the literature was conducted to examine: (a) link between environmental hazards and falls, (b) efficacy of home-modification interventions, and (c) role of the physical environment in falls of people with cognitive impairment. RESULTS: A strong link between environmental hazards and the risk of falls has yet to be established. A conceptual framework is presented that proposes that an individual's risk for falls can be determined by the interaction of three main factors: mobility, risk-taking behaviour, and physical environment. IMPLICATIONS: Environmental interventions should be combined with other interventions such as exercise programs and education. PMID- 18510253 TI - Giving Youth a Voice (GYV): a measure of youths' perceptions of the client centredness of rehabilitation services. AB - BACKGROUND: Although client-centred care is regarded as the optimum way of delivering health care, there is currently no method to measure the client centredness of services for youth with disabilities. PURPOSE: To develop a measure of youths' perceptions of the client-centredness of health care services in rehabilitation. METHODS: The Giving Youth a Voice (GYV) questionnaire was adapted from the Measure of Processes of Care, a measure of caregiving from the perspective of parents. Items for a youth version of the questionnaire were generated from focus groups with youths who had received rehabilitation services. Content analysis of the groups yielded four themes, which became the subscales for the new measure. FINDINGS: GYV, a 56-item measure, has good internal reliability and test-retest reliability. Convergent validity was estimated through correlations with the Client Satisfaction Questionnaire. IMPLICATIONS: GYV provides an opportunity for youth with disabilities to have a voice about the rehabilitation services they receive. PMID- 18510254 TI - Factors influencing therapists' interventions for children with learning difficulties. AB - BACKGROUND: Occupational therapists use a range of intervention approaches with children with learning difficulties. PURPOSE: This study aimed to identify factors influencing therapists' clinical decisions when choosing and combining these interventions. METHODS: Seven occupational therapists participated. Data were collected through in-depth interviews, short questionnaires, and therapy observations and analysed thematically. FINDINGS: Therapists used a combination of child, therapist, and service-related factors to individually tailor a combination of intervention techniques drawn from different theoretical approaches. Therapists also used embedded practices with all children to engage and motivate them, promote task mastery, encourage application of strategies at home and school, and evolve intervention through ongoing evaluation. IMPLICATIONS: The child and family's home and school context and, to a lesser extent, the therapist's past experience and service context are primary factors considered when choosing interventions. Therapists continually move between theoretical and practical levels of thinking to best meet each child's needs. PMID- 18510255 TI - [Analysis of the concept of social participation: definitions, illustration, dimensions of activity and indicators]. AB - BACKGROUND: Social participation is an integral part of human life. This concept has appeared in recent health literature and is considered one of the main goals of rehabilitation. PURPOSE: The purpose of this article is to clarify the concept of social participation and its applicability in mental health. The concept of social participation is distinguished from other related concepts and measurement tools assessing social participation are examined. METHODS: The analysis was done according to the Walker and Avant (1995) method. FINDINGS: The analysis identified three attributes of social participation: 7) participation implies an action from the individual; 2) this action contributes to others; 3) personal and societal dimensions need to be considered. Based on the examination of 77 measurement tools, there appears to be operational consensus that social participation is the realization of activities. The most often used indicators include amount and frequency of activities. IMPLICATIONS: The Assessment of Life Habits and the Participation Measure for Post-Acute Care appear to be the most comprehensive assessments available to measure social participation in mental health that are currently available. PMID- 18510256 TI - [Supporting doctors with mental health problems]. PMID- 18510257 TI - Short-lived doctors. 1908. PMID- 18510258 TI - Improving pain detection in older patients. PMID- 18510259 TI - Infestations. PMID- 18510260 TI - Remembering ABCDE in general practice. PMID- 18510261 TI - [Genetic markers and personality traits in eating disorders--preliminary results]. AB - INTRODUCTION: The multidimensional approach of the ethiopathogenesis of eating disorders include the genetic, biologic, psychosocial effects, and premorbid personality markers. AIM/METHOD: To determine the potential relation between genetic and personality trate and state factors, and also to investigate the connection of clinical symptoms and diagnostic subgroups. The serotonin transporter gene (VNTR) polymorphism was investigated by polymerase chain reaction (PCR) technology, the personality factors were determined by the Temperament and Character Inventory (TCI) inventory. RESULTS: Among patients with bulimia nervosa (BN) the short allele of serotonin transporter gene was more frequent than in anorexia nervosa (78% vs. 67%), and in both groups it was more common than in the general population (43%). Patients with anorexia nervosa (AN) have higher scores in the scale of harm avoidance (61,2 vs. 51.4), but in bulimia nervosa the novelty seeking (54.5 vs. 44.2) and the reward dependence factors (53.2 vs. 46.5) were more significant. In the self-directedness (BN: 42.7; AN: 44.3) and the cooperativeness scales (BN: 51.2; AN: 44.6) both groups show lower scores, which could implicate personality disorder in the background of the eating disorders. Anorexic patients with the 10 allele show similar personality factors like patients with bulimia nervosa, while with the 12 allele (homozygotes), their factors were more likely the factors of patients with classic anorexic symptoms. CONCLUSION: Our data strength the role of specific personality factors in the background of the symptoms of eating disorders. Among patients with bulimia nervosa the 10 allele were more frequent, which could indicate the role of the serotonin system in developing eating disorders. Two subgroups were differentiated among patients with anorexia nervosa in relation with personality factors; the factors of patients with the 12 allele homozygotes were similar to the classical factors of anorexia nervosa, while patients with the 10 allele were like bulimic patients. Our results could improve our knowledge with newer aspects concerning the etiology of eating disorders, that might be used in broadening our preventive and therapeutic facilities in the future. PMID- 18510262 TI - [Markers of cognitive vulnerability in major depression]. AB - Neuropsychological deficits have been reported in patients with major depressive disorder in a lot of lectures. The aim of the present study was to measure the neurocognitive functions of major depressive patients and healthy controls, and identify some vulnerability marker of the disease. Seventy-one patients with major depressive disorder were compared to thirty healthy control subjects on tasks from Trail Making Test, Stroop Test, Rey Verbal Auditory Learning Test and Rey-Osterreich Figure Test. Patients with depression relative to controls were significantly impaired on tasks of attention, executive function, memory and psychomotor speed, but not in visuo-spatial function. These findings suggest deficits in cognitive function as, attention, visual and verbal memory and learning, as vulnerability marker for major depressive disorder. The second important finding, the better result of the visual perception and short memory in depressed patient, was caused by intact hippocampal function. The functional importance of this result is the connection of the neuronal network and signs of depression. PMID- 18510263 TI - [Administration of once-daily extended release quetiapine in schizophrenic disorders]. AB - This article reviews the published clinical data on schizophrenic patients managed with the new formulation of quetiapine, the once-daily extended release quetiapine fumarate (quetiapine XR). Quetiapine XR has been developed to reduce the frequency of quetiapine dosing by introducing once-daily administration and to simplify the treatment initiation schedule. Quetiapine XR (400 to 800 mg/day) was effective versus placebo across a broad range of symptom domains in acute schizophrenia and was as well tolerated as the immediate release (IR) formulation. Rapid dose escalation of quetiapine XR (300 mg on day 1,600 mg on day 2, and 800 mg on day 3) was also well tolerated, with a therapeutically effective dose reached by day 2. Clinically stable patients with schizophrenia receiving quetiapine IR (400-800 mg/day) can be switched to an equivalent once daily dose of quetiapine XR (400-800 mg/day once-daily) without clinical deterioration or compromise in tolerability. Evidence from a clinical trial has shown that patients with schizophrenia who had a history of unsatisfactory treatment (tolerability or efficacy) on typical or atypical antipsychotic experienced improved efficacy and clinical benefit when switched to quetiapine XR. Once-daily quetiapine XR (400-800 mg/nap) was effective compared with placebo in preventing relapse in patients with clinically stable schizophrenia, and was well tolerated during long-term use. Patients could be switched from their ongoing antipsychotic to quetiapine XR within 4 days without compromising efficacy, enabling a dose of 600 mg/day and 800 mg/day to be reached by Day 2 and Day 3 respectively. This new, once-daily formulation of quetiapine offers psychiatrists and patients valuable new treatment options for the short and long term treatment of schizophrenia. PMID- 18510264 TI - [Mental retardation in childhood. The prevalence of informative morphogenetic variants]. AB - The authors observed the prevalence of informative morphogenetic variants among mentally retarded children. In their retrospective examination of a 4-years period, they included 135 hospitalized children and analysed their clinical epidemiological and dysmorphic profiles. In their descriptive analysis they found ear abnormalities and abnormal head circumference as the most common dysmorphic sings among mentally retarded children, and they emphasize the highly different importance and predictive-diagnostic value of minor malformations and phenogenetic variants. PMID- 18510265 TI - DMT at fifty. AB - The steps taken for the discovery of the hallucinogenic effects of N.N-Dimethyl tryptamine (DMT) is described. DMT had a difficult first 50 years in medical research primarily for legal reasons as it was classified as one of the "drugs of abuse" by authorities in the USA and by the World Health Organization. It has not proved to be a "schizotoxin" as it was first suspected, but the book is not closed on its potential role in some other, high level function as an endogenous neuromodulator. Further clinical work may even substantiate its usefulness in therapeutic application, such as an adjunct to psychotherapy, perhaps not by itself, but in a modified form, or in combination with other substances. PMID- 18510266 TI - [Successful clozapine treatment of primary polydipsia associated with hyponatraemia in a schizophrenic patient. A case report]. AB - Polydipsia is the intake of more than 3-4 litres of fluids per day. Primary polydipsia (PP) occurs when excessive fluid intake cannot be explained by an identified medical condition. PP has a prevalence varying between 6% and 20% in the population of chronically hospitalized psychiatric patients. Hyponatraemia- sometimes with severe somatic consequences--developing in 25-86% of these patients. We discuss the case of a schizophrenic patient who had polydipsia, polyuria and hyponatremia without any known medical conditions in the etiological background of these symptoms. In accordance with data of literature, clozapine medication was effective in the treatment of this severe condition. PMID- 18510267 TI - [Latex exposure and gloves' use in health settings: old and new issues]. AB - The diffusion of the "universal precautions", promoted in 1987 by the Center for Disease Control and Prevention, dramatically increased the use of latex glove in health care settings for protection against the HIV and HBV. The increased demand caused an increased production of gloves, a reduction in processing time and a different chemical treatment of rubber trees which lowered the glove quality, that means high levels of antigens and high powder content. This situation caused an increase of frequency of allergic (type I and type IV) and irritant reactions to latex gloves in health care workers. Recommendations and guidelines for the prevention of latex allergy in health care settings were available from 1995 both in Italy and in other countries. When properly applied, the preventive effectiveness of these suggestions has been demonstrated, but unfortunately the guidelines are not widely adopted as we could think and the frequency of latex allergy and sensitization in health care workers is not as reduced as it could be expect. This could be due also to the fact that some practical issues are still open, for example: the possibility of assessing the real glove quality and the reliability of information provided by manufacturers that often are inaccurate and incomplete; the necessity to update the regulations in force to higher quality standards; the availability of procedures for the selection, purchase and use of gloves in relation to specific tasks within health care setting; the involvement of occupational physicians in the management of these procedures; clear indications of limit values for extractable latex allergens in medical and common latex devices; the assessment of the real protective efficacy against chemicals and biological agents of new synthetic rubber gloves. An Italian working group of occupational health professionals, involved for a long time in the management of glove and latex related problems in health care settings,finalized a consensus document with practical suggestions and possibly answers to the above mentioned questions. PMID- 18510268 TI - [Old and new types of sanitary gloves: what has improved?]. AB - BACKGROUND: During the eighties a large increase in latex gloves production was observed because of the high demand of gloves in health care settings. In this period a low compliance to minimal quality standard was detected and the poor glove quality was associated with an increase of both irritant and allergic glove related diseases. Since the second half of nineties health care workers and manufacturers paid more attention to these problems and a trend to a gradual, even if slow, quality improvement was observed. Most frequently powder-free gloves and synthetic gloves were offered on the market. OBJECTIVES: The aim of this study was to highlight what has improved about materials and types of sanitary gloves during the last ten years. METHODS: The information are based on a review of the scientific literature and practical experiences. RESULTS AND CONCLUSIONS: Today a large selection of gloves made of different materials are available and they should be addressed to specific tasks. The review of the scientific literature and the analysis of many technical sheets provided by the manufacturers pointed out a trend to a better latex gloves quality (less chemical additives and generally a lower total protein content); sometimes data about a lower extractable latex allergens content are also available. Unfortunately detailed information on glove composition are not usually provided by the manufacturers; purchasers should require the manufacturing company to give comprehensive information and verify their reliability. Moreover the regulation in force should be adapted to higher quality standards. Powder-free and synthetic gloves consumption has improved but the use of synthetic rubber gloves should be further enhanced since some materials (e.g. neoprene and nitrile rubber) have a good biocompatibility and seem to have physical properties and protective efficacy similar to latex. Moreover allergic reactions to synthetic gloves (some chemical additives) are only occasional. PMID- 18510269 TI - [Gloves as medical devices or individual protection devices: current regulations and correct use in hospital environments]. AB - BACKGROUND: Hospital work consists of the care and assistance of patients, who therefore constitute another group of individuals besides the workers, which is a completely different situation compared to other workplaces in manufacturing industries. OBJECTIVES: Health professionals must know how to use the right devices to protect both the patients and themselves. In the case of protection of the hands, health professionals must also know when to use gloves as "individual protection gloves" and when to use gloves as "medical devices", and comply with both Italian and European Community regulations. METHODS AND RESULTS: The Hospital of Vimercate (Milan) has drawn up technicalprocedures regarding the various types of gloves used in hospitals, which provide health care workers with accurate information on regulations and protocols, prevention guidelines, safety management recommendations, purchasing specifications, quality controls, correct uses of gloves and devices. PMID- 18510270 TI - [Criteria for choice and purchase of gloves: quality or cheapness? The role of the Occupational Health Physician]. AB - BACKGROUND: The choice of gloves in the healthcare settings is very important because of the high biological and chemical risks present in these workplaces. In order to rationalize and optimize this choice we must balance cost, quality, security and comfort. OBJECTIVES: The aim of this study was to analyse the rules in force and to point out the relevant role of the Occupational Health Physician in the right choice and purchase of sanitary gloves. METHODS: We reviewed the rules in force and the most relevant studies on these topics. RESULTS AND CONCLUSIONS: The regulations in force provide that the manufacturers must perform tests to supply evidence for the quality of the products but they do not indicate which analytical method should be used and they do not require that the results are reported in the technical sheets. Thus the manufacturers have only to declare to be "in accordance with the rules". Therefore purchasers should require the manufacturing companies to give detailed information and verify their reliability. Moreover rules could be adapted to higher quality standards. The Occupational Health Physician should suggest the purchase of gloves with high biocompatibility, assuring the protection from the risks of specific tasks and suitable for preventing the onset of new glove-related diseases and the relapses in workers with already diagnosed occupational diseases. PMID- 18510271 TI - [Chemical resistance of latex gloves]. AB - Dermal chemical risk is represented by the chemicals with irritant, caustic and sensitization properties or that can produce toxic effects after penetration through the skin. During the last few years the interest for the effective protecting ability of gloves progressively is increased in parallel with their use. Penetration of chemicals through the protecting material does not always turn out visible. That has led to the development of test standardizes. In 1989 the EC adopted two Directives in the field of the characteristics of protection devices defining certification procedures (89/686/EEC) and characteristics demanded for the use of protection devices at the workplace (89/656/EEC). When the European Standards are accepted from the CEN they exist in rough draft shape (prEN), losing the prefixed one pr- when they are approved by all the EC member Countries. A number of EN for protection gloves have been proposed by CEN/TC 162 regarding methods for testing the resistance to penetration/permeation of chemicals and other characteristics. However in these protocols some limitations were found, since not necessarily they represent the real using conditions. In general an ideal protecting material against the chemicals does not exist because what is effective to a specific compound does not effectively protect from an other. Latex is not commonly considered a very chemical resistant material and therefore its use in such sense is limited However the low cost and the spread of latex gloves in particular in Public Health often make them easy available at workplace and therefore used in several situations. PMID- 18510272 TI - [Latex and synthetic polymers gloves: protective effectiveness against biological risk]. AB - BACKGROUND: The quick evolution of knowledge about latex and, especially, its effectiveness as protection against biological risks on one hand, and, on the other hand, as trigger of cutaneous, respiratory and sistemic allergic diseases, required a marked distinction between conditions in which latex is necessary and conditions in which latex could be substituted by syntetic polymers. OBJECTIVES: The aim of this study is to verify protective effectiveness of gloves made of syntetic polymers against biological risks compared with latex gloves which until now have been considered as the best protection. METHODS: Literature review. RESULTS: The most recent studies seem to consider neoprene gloves as the safest protection to replace latex in working activities with high biological risk exposure. CONCLUSION: The literature review marked the lack of studies exclusively focused on glove permeability towards biological agents. It seems therefore necessary to investigate this topic to validate results and to guarantee a suitable protection to workers daily exposed to biological risk. PMID- 18510273 TI - [Epidemiology of latex allergy in healthcare workers]. AB - BACKGROUND: Natural rubber latex is a recognized allergen and health care workers have an increased risk of sensitisation and allergic symptoms to latex. OBJECTIVES: The aim of the study was to evaluate prevalence and incidence of latex sensitisation and symptoms in health care workers. METHODS: We analyzed the reported data from the scientific literature and from the follow-up study in health care workers in Trieste Hospitals carrying on since 1999. RESULTS: Latex allergy and sensitisation are significantly higher in health care workers respect to the general population with symptoms, such as urticaria, rhinoconjunctivis and asthma, ranging between 4 and 5.6%. Latex sensitisation evaluated by skin prick test is ranging between 6 to 7.8%. All studies done demonstrated the reduction of symptoms and incidence sensitisations after the changeover to a powder- free environment and the avoidance of unnecessary glove use. CONCLUSIONS: Health care workers have an increased risk of sensitisation and allergic symptoms to latex but the use of un-powdered latex gloves by all workers and use of non-latex gloves by sensitized subjects can reduce symptoms and prevent new sensitisation. Nevetheless the use of no-latex gloves when avaiable has to be the best choice from the preventive point of view. PMID- 18510274 TI - [Natural latex allergy. Patient management: from clinic to prevention. A review]. AB - The widespread use of latex devices has been followed, in the last 25 years, by an increase in IgE mediated sensitization. The clinical manifestations of latex allergy affect the skin (urticaria and angioneurotic oedema), the lower and the upper respiratory tracts (rhinoconjunctivitis, asthma and glottis oedema), and the cardiovascular system (anaphylaxis). There is also an anaphylactic risk during surgery and invasive diagnostic procedures. Vegetable food cross-reacts with latex so that more than half of the patients show specific IgE against some food. Further than traditional groups at risk, as health care workers, other work categories have to be protected, because of the inappropriate use of latex gloves (food or drug industry workers, mechanics, panel beaters and so on). Recently the latex most important allergenic fractions have been characterized and recombinant allergens are now available. The recombinant allergens allow a better standardization of the extracts for diagnostic use, the production of safer extracts for immunotherapy as well as a more accurate evaluation of food cross reactions. The recombinant allergens will allow a more accurate dosage of latex concentrations in air and in objects and, in future, to establish threshold limit values. The main aims of prevention are the replacement of latex with alternative elastomers, the reduction of work and extra work exposure and an efficient health survey in working environment. The use of latex gloves and devices among general population has to be discouraged. Specific immunotherapy has to be considered a second choice and restricted to highly qualified workers in order to realize a rehabilitation to their previous jobs. The actually obtained protection must be verified. PMID- 18510275 TI - [Monitoring latex risk: detection of airborne and glove allergens]. AB - BACKGROUND: Latex is a relevant occupational and environmental allergen, strongly related to the extensive use of natural rubber products. OBJECTIVES: Threshold Limit Values have to be identified, as well as biocompatible materials in order to avoid sensitization or appearance of allergic symptoms. METHODS: In this paper we consider the main methods, which have been used to detecting latex allergens for environmental monitoring of airborne and latex products. RESULTS: We report our experience in such afield, and our approach to the latex problem, suggesting that quantification of allergens, which is currently applicable according to well standardized methods, should be adopted by manufacturers, agency and consumer organization. PMID- 18510276 TI - [Italian and international guidelines on latex allergy: revision and comparison]. AB - BACKGROUND: Latex allergy is an important medical issue, particularly for some occupationally exposed categories (healthcare personnel). The increase in the number of cases reported, occurred as a consequence of the diffusion of latex use, prompted several scientific and institutional organisms, both on a local and international level, to create guidelines for risk management in healthcare environment. OBJECTIVES: To review and compare, more than ten years after the first official guidelines, the directives on latex allergy currently used in different countries and in the Italian regions. METHODS: We used the references contained in the relevant literature and in the regulations concerning prevention of latex allergy. RESULTS: Epidemiologic data, together with the recent decrease of the number of papers on latex allergy, suggest a relevant, but still unsatisfactory reduction of the problem. CONCLUSIONS: In the light of a recent study performed by National Health Service (NHS) in UK, we point out the need for a critical evaluation of the real application of the existing guidelines, which are often formally present but left unattended in the daily hospital practice. PMID- 18510277 TI - [Latex allergy. Preventive measures in health care workers: a comparison of Italian experiences]. AB - BACKGROUND: We have performed a transversal study in different Italian Regions to underline the problem related to natural latex allergy. OBJECTIVES: The aim of the study was to determine the glove use on the basis of the materials gloves were made of, to identify the departments and the qualifications mainly interested in the latex allergy problem, to check the required features for gloves and other rubber devices at the moment of the purchase, as well as the presence of guidelines, of training and information activities for health care workers and procedures for sensitized patients. METHODS: We sent a questionnaire to the local public Health Authorities (HA) in different Italian Regions. RESULTS: Number and regional distribution of the answers received back did not have statistical significance. Anyway, from the elaboration of the data it has been possible to obtain some interesting observations on both a theoretical and an operational level. The results showed: a) the level of interest and attention to the latex allergy problem has increased a lot recently; b) there is a high use of latex gloves (>50% of HA), in particular in surgical and intensive care departments; c) there are differences on an operational level between HA, also between HA that are in the same Italian Region. It resulted a widespread delay in the replacement of latex devices (e.g. urinary and intravenous catheters) with devices made of alternative materials. However, we found a growing attention towards the procedures related to the purchase of gloves, towards the adoption of guidelines about the use of gloves and towards the training and the information of health care workers. CONCLUSIONS: From this study it has emerged clear the need of Italian HA to focus on the prevention of latex allergy, goal already obtained in a few, almost isolated, realities. PMID- 18510278 TI - [Prevention of building site accidents in Umbria during reconstruction after an earthquake: the experience of one local health unit]. AB - BACKGROUND AND OBJECTIVES: Building site related accidents are so frequent and are associated with such serious consequences that they constitute a major "health emergency" in Italy where dangerous work conditions in the construction industry often derive from excessive financial, organisational and time pressures. After the 1997 earthquake in Umbria an extensive re-building programme led to a concentration of innumerable construction firms, sub-tendering practices and the use of poorly skilled workforces who were often employed in the black economy. At the same time, SPSAL (Prevention Service for Occupational Settings) activity needed to be intensified in Perugia Health Unit No 2 in the District of Assisi, which had been devastated by the earthquake. DISCUSSION AND CONCLUSIONS: To monitor construction site development, related accidents and injuries, and surveillance activity, data were collected from building site notifications on the basis of Article 11, Law 494/96, outcomes of inspections, including sanctions, and INAIL (National Insurances) certificates of work-related injuries. Construction notifications showed building sites increased in number from the 200 sites/year before the earthquake to almost 1400 per year at maximum reconstruction activity. Inspections and surveillance also increased to over 350 inspections/year in 250 sites/year. About 600 firms were monitored and sanctions increased Sanctions mainly referred to high-level work, scaffolding, protection against falls from heights, and DPI. (Personal Protective Equipments). Accidents increased from 150/year before the earthquake to about 300/year. The annual incidence, a rough indicator of prevention, dropped, indicating good quality prevention strategies were in force. The drop in the annual incidence index would not have been evident without concomitant monitoring of notifications and certificates of work-related injuries. These two databases are invaluable tools when investigating changes in the reference stan dard of the construction industry and assessing the efficacy of implemented prevention programmes. PMID- 18510279 TI - [Continuing medical education and accreditation for excellence set up by the Italian Society of Occupational Health and Industrial Hygiene (SIMLII): results of a survey among members]. AB - BACKGROUND: The SIMLII set up a program of continuous training and accreditation of postgraduate specialists in Occupational Health in 2000, before continuous medical training (ECM) was formally introduced. Therefore, especially over the past few years, SIMLII has involved its membership to an ever increasing extent, in an attempt to understand their expectations as regards the scientific community and as regards a continuously evolving discipline that is subject to continuous changes in educational requirements. OBJECTIVES AND CONCLUSIONS: The self-administered questionnaire in a survey carried out in 2005 by the National Institute of Occupational Safety and Prevention (ISPESL), in collaboration with SLMLII, provided data that, besides identifying the type of activity of the membership, attempted to assess members' compliance with the society's initiatives, with special reference to the Programme for continuous training and accreditation of postgraduates in Occupational Health and the issue of Guidelines. PMID- 18510280 TI - Focus on hypertriglyceridemia: improving patient outcomes. PMID- 18510281 TI - Pathogenesis of CHD: the role of elevated triglycerides. PMID- 18510282 TI - Hypertriglyceridemia and cardiovascular disease management. PMID- 18510283 TI - Tailoring patient care. PMID- 18510284 TI - Coordination complexes exhibiting anion...pi interactions: synthesis, structure, and theoretical studies. AB - The polydentate ligand 2,4,6-tris(dipyridin-2-ylamino)-1,3,5-triazine (dpyatriz) in combination with the Cu(ClO 4) 2/CuX 2 salt mixtures (X (-) = Cl (-), Br (-), or N 3 (-)) leads to the formation of molecular coordination aggregates with formulas [Cu 3Cl 3(dpyatriz) 2](ClO 4) 3 ( 2), [Cu 3Br 3(dpyatriz) 2](ClO 4) 3 ( 3), and [Cu 4(N 3) 4(dpyatriz) 2(DMF) 4(ClO 4) 2](ClO 4) 2 ( 4). These complexes consist of two dpyatriz ligands bridged via coordination to Cu (II) and disposed either face-to-face in an eclipsed manner ( 2 and 3) or parallel and mutually shifted in one direction. The copper ions complete their coordination positions with Cl (-) ( 2), Br (-) ( 3), or N 3 (-), ClO 4 (-), and N, N-dimethylformamide (DMF) ( 4) ligands. All complexes crystallize together with noncoordinate ClO 4 ( ) groups that display anion...pi interactions with the triazine rings. These interactions have been studied by means of high level ab initio calculations and the MIPp partition scheme. These calculations have proven the ClO 4 (-)...[C 3N 3] interactions to be favorable and have revealed a synergistic effect from the combined occurrence of pi-pi stacking of triazine rings and the interaction of these moieties with perchlorate ions, as observed in the experimental systems. PMID- 18510285 TI - Solid-state molecular rotators of anilinium and adamantylammonium in [Ni(dmit)2]( ) salts with diverse magnetic properties. AB - Supramolecular rotators of hydrogen-bonding assemblies between anilinium (Ph-NH 3 (+)) or adamantylammonium (AD-NH 3 (+)) and dibenzo[18]crown-6 (DB[18]crown-6) or meso-dicyclohexano[18]crown-6 (DCH[18]crown-6) were introduced into [Ni(dmit) 2] salts (dmit (2-) is 2-thioxo-1,3-dithiole-4,5-dithiolate). The ammonium moieties of Ph-NH 3 (+) and AD-NH 3 (+) cations were interacted through N-H (+) approximately O hydrogen bonding with the six oxygen atoms of crown ethers, forming 1:1 supramolecular rotator-stator structures. X-ray crystal-structure analyses revealed a jackknife-shaped conformation of DB[18]crown-6, in which two benzene rings were twisted along the same direction, in (Ph-NH 3 (+))(DB[18]crown 6)[Ni(dmit) 2] (-) ( 1) and (AD-NH 3 (+))(DB[18]crown-6)[Ni(dmit) 2] (-) ( 3), whereas the conformational flexibility of two dicyclohexyl rings was observed in (Ph-NH 3 (+))(DCH[18]crown-6)[Ni(dmit) 2] (-) ( 2) and (AD-NH 3 (+))(DCH[18]crown 6)[Ni(dmit) 2] (-) ( 4). Sufficient space for the molecular rotation of the adamantyl group was achieved in the crystals of salts 3 and 4, whereas the rotation of the phenyl group in salts 1 and 2 was rather restricted by the nearest neighboring molecules. The rotation of the adamantyl group in salts 3 and 4 was evidenced from the temperature-dependent wide-line (1)H NMR spectra, dielectric properties, and X-ray crystal structure analysis. ab initio calculations showed that the potential energy barriers for the rotations of adamantyl groups in salts 3 (Delta E approximately 18 kJmol (-1)) and 4 (Delta E approximately 15 kJmol (-1)) were similar to those of ethane ( approximately 12 kJmol (-1)) and butane (17-25 kJmol (-1)) around the C-C single bond, which were 1 order of magnitude smaller than those of phenyl groups in salts 1 (Delta E approximately 180 kJmol (-1)) and 2 (Delta E approximately 340 kJmol (-1)). 1D or 2D [Ni(dmit) 2] (-) anion arrangements were observed in the crystals according to the shape of crown ether derivatives. The 2D weak intermolecular interactions between [Ni(dmit) 2] (-) anions in salts 1 and 3 led to Curie-Weiss behavior with weak antiferromagnetic interaction, whereas 1D interactions through lateral sulfur-sulfur atomic contacts between [Ni(dmit) 2] (-) anions were observed in salts 2 and 4, whose magnetic behaviors were dictated by ferromagnetic (salt 2) and singlet-triplet (salt 4) intermolecular magnetic interactions, respectively. PMID- 18510286 TI - Reactions of the Re(CO)3(H2O)3(+) synthon with monodentate ligands under aqueous conditions. AB - The reactions of ammonia, pyridine (py), N-methyl imidazole (N-MeIm), tetrahydrothiophene (tht), and piperidine (pip) with Re(CO) 3(H 2O) 3 (+), 1 ( + ), were investigated employing aqueous conditions under atmospheric dioxygen. The reaction of [ 1]Br in aqueous ammonia led to [Re(CO) 3(NH 3) 3]Br ([ 2]Br) as the only product isolated. For the aqueous reactions of [ 1]Br with py, N-MeIm, and tht, mixtures of products are formed because of competition between the bromide and added ligand, even when the ligand is present in excess. Substitution of the PF 6 (-) anion for Br (-) leads to the clean formation of [Re(CO) 3L 3][PF 6] ([ 3][PF 6]-[ 5][PF 6]) for py, N-MeIm, and tht, respectively, as the only products observed. Reaction of [ 1][PF 6] with pip produces the dimeric species, (pip)(CO) 3Re(micro-OH) 2Re(CO) 3(pip), 6. Reactions of [ 1]Br were also performed in methanol for comparison purposes. The reaction with pip in this solvent led to the analogous dimer, (pip)(CO) 3Re(micro-OMe) 2Re(CO) 3(pip), 7; however, reactions with py, N-MeIm, and tht gave Re(CO) 3L 2Br, 8- 10, respectively, as the only products. The crystal structures of compounds [ 2]Br- 10 are reported. PMID- 18510287 TI - Hard and soft acids and bases: atoms and atomic ions. AB - The structural origin of hard-soft behavior in atomic acids and bases has been explored using a simple orbital model. The Pearson principle of hard and soft acids and bases has been taken to be the defining statement about hard-soft behavior and as a definition of chemical hardness. There are a number of conditions that are imposed on any candidate structure and associated property by the Pearson principle, which have been exploited. The Pearson principle itself has been used to generate a thermodynamically based scale of relative hardness and softness for acids and bases (operational chemical hardness), and a modified Slater model has been used to discern the electronic origin of hard-soft behavior. Whereas chemical hardness is a chemical property of an acid or base and the operational chemical hardness is an experimental measure of it, the absolute hardness is a physical property of an atom or molecule. A critical examination of chemical hardness, which has been based on a more rigorous application of the Pearson principle and the availability of quantitative measures of chemical hardness, suggests that the origin of hard-soft behavior for both acids and bases resides in the relaxation of the electrons not undergoing transfer during the acid-base interaction. Furthermore, the results suggest that the absolute hardness should not be taken as synonymous with chemical hardness but that the relationship is somewhat more complex. Finally, this work provides additional groundwork for a better understanding of chemical hardness that will inform the understanding of hardness in molecules. PMID- 18510288 TI - New polydentate ligand and catalytic properties of the corresponding ruthenium complex during sulfoxidation and alkene epoxidation. AB - Bis(diimine)-ruthenium complexes constitute a class of catalysts with good activity for oxidation reactions, such as sulfoxidation and epoxidation. The synthesis and the full characterization of a new ruthenium complex bearing an original pentadentate ligand (L5pyr for 2,6-bis-(6-ethyl-2,2'-bipyridyl) pyridine) is reported. Comparison of its activity with regard to[Ru(bpy)2(CH3CN)2](2+) and [Ru(bpy)2(py)(CH3CN)](2+) during alkene and sulfide oxidation allowed us to conclude that the addition of a fifth pyridine ligand in the coordination sphere improves the efficiency of the catalyst. Moreover, under these oxidation conditions a hydroxylation of the ligand L5pyr led to a better activity than its analogue [Ru(bpy)2(py)(CH3CN)](2+), especially during epoxidation of alkenes by PhI(OAc)2. PMID- 18510289 TI - Structures of [(n-C4H9)2NH2]2Cd9Cl20.2H2O and [Cu(C14H24N4)]2Cu13Cl30(H2O)2.xH2O: perforated layer structures based on the CdCl2 layer network. AB - The crystal structures are reported for two compounds containing novel perforated layer structures, [DBA] 2Cd 9Cl 20.2H 2O and [Cu(TIM)] 2Cu 13Cl 30(H 2O) 2. xH 2O, where [DBA] (+) = di n-butylammonium and TIM = 2,3,9,10-tetramethyl-1,3,8,10 tetraenecyclo- 1,4,8,11-tetraazatetradecane. In the former compound, single Cd (2+) ions are excised from the parent CdCl 2 layers, with water molecules hydrogen bonded to chloride ions on both sides of the excision. Lattice stability is provided by the DBA (+) cations, which have an all-trans conformation. These lie between the layers, hydrogen bonding to the adjacent [Cd 9Cl 20(H 2O) 2] n (2 n- ) sheets. In the copper compound, the modification of the parent CuCl 2 structure is much more complex. In this compound, [Cu 2Cl 2] (2+) moieties are excised in a regular fashion. In addition, at 50% of the Cu1 sites, CuCl 2 species are replaced by pairs of water molecules in a random fashion. The Cu(TIM) (2+) cations bridge the layers via the formation of two semicoordinate bonds to chloride ions at the edge of the [Cu 2Cl 2] (2+) excision sites of adjacent layers. PMID- 18510290 TI - (Alpha-diimine)chromium complexes: molecular and electronic structures; a combined experimental and density functional theoretical study. AB - Dark brown crystals of [Cr( (1)L) 2] ( 1) were obtained from the reaction of [Cr (III)(acac) 3] (acac (-) = 2,4-pentanedionate) with 2 equiv of 2-methyl-1,4 bis(2,6-dimethylphenyl)-1,4-diaza-1,3-butadiene ( (1)L) and 3 equiv of sodium in tetrahydrofuran (thf) under an Ar atmosphere. Complex 1 possesses an S = 1 ground state, which is attained via intramolecular antiferromagnetic coupling between a high-spin Cr (II) ion ( S Cr = 2) and two anionic alpha-diiminato(1-) ligand pi radicals ( (1)L (*)) (1-). The molecular structure of 1 exhibits a distorted tetrahedral, nearly square-planar geometry. The average C-N imine bond length at 1.346 A is characteristic for the pi radical anion ( (1)L (*)) (1-), and therefore, the electronic structure of 1 is best described as [Cr (II)( (1)L (*)) 2]. This has been confirmed by broken symmetry density functional theoretical calculations BS(4,2) (DFT) at the B3LYP level. The reaction of [Cr (III)(acac) 3] with 1 equiv of 2,3-dimethyl-1,4-bis(2,6-diisopropylphenyl)-1,4-diaza-1,3 butadiene ( (2)L) and 1 equiv of Na in thf under Ar yields red-brown crystals of [Cr (III)( (2)L (*))(acac) 2] ( 2) ( S = 1). The oxidation of 2 with 1 equiv of Fc(PF 6) (Fc (+) = ferrocenium) in CH 2Cl 2 affords crystals of [Cr (III)( (2)L (ox))(acac) 2](PF 6) ( 3) ( S = (3)/ 2). The crystal structure determinations of 2 and 3 revealed that 2 contains a neutral, octahedral Cr (III) species [Cr (III)( (2)L (*))(acac) 2], whereas in 3 the ligand is oxidized, yielding an octahedral monocation [Cr (III)( (2)L (ox))(acac) 2] (+). These electronic structures have been confirmed by DFT calculations. PMID- 18510291 TI - Cyclodimers versus cyclotrimers via solvent or temperature effects on metallacyclization. AB - The reaction of (COD)PdCl 2 (COD = 1,5-cyclooctadiene) with bis(3 pyridyl)methylphenylsilane (L) in acetone affords single crystals consisting of cyclodimers, [PdCl 2(L)] 2, whereas the reaction in a mixture of dichloromethane and ethanol yields amorphous spheres consisting of cyclotrimers, [PdCl 2(L)] 3. The interconversion and morphology control between the crystals and the microspheres can be explained by the difference in flexibility between the cyclodimer and cyclotrimer. PMID- 18510292 TI - Novel Michael addition products of bis(amino acidato)metal(II) complexes: synthesis, characterization, dye degradation, and oxidation properties. AB - Michael addition reactions of bis(amino acidato)metal(II) complexes (metal = copper, nickel, zinc; amino acid = glycine, dl-alanine, l-alanine) with acrylonitrile have been carried out under various experimental conditions in the absence of a base, resulting in mono- and disubstituted products in high yield, including partially hydrolyzed products. A reaction mechanism for the Michael addition on the nitrogen atom of the coordinated amino acid moiety, replacing the amino hydrogen atom(s), is proposed. All of the products have been characterized by Fourier transform infrared spectroscopy, electron paramagnetic resonance spectra, and elemental and electrochemical analyses. The single-crystal structures of bis( N-cyanoethylglycinato)copper(II) monohydrate ( 1a), diaquabis( N-cyanoethylglycinato)nickel(II), aquabis( N, N-dicyanoethylglycinato)copper(II) ( 2a), and bis[( N-propionamido- N-cyanoethyl)glycinato]copper(II) dihydrate ( 4a) have been confirmed by X-ray diffraction techniques. The products 1a, 2a, 4a, and bis( N-propionamidoglycinato)copper(II) monohydrate ( 3a) have been used as catalysts for the degradation of a phenol red dye and mild oxidation of various organic substrates in the presence of hydrogen peroxide. The monosubstituted complexes have been found to catalyze the reactions to a greater extent than the disubstituted complexes. PMID- 18510293 TI - Lanthanide-centered covalently bonded hybrids through sulfide linkage: molecular assembly, physical characterization, and photoluminescence. AB - A series of novel photoactive lanthanide (europium, terbium, dysprosium, samarium) hybrid materials with organic parts covalently bonded to inorganic parts via sulfide linkage have been assembled by the sol-gel process. The organic parts as molecular bridge are obtained from the functionalized thiosalicylic acids by five silane crosslinking reagents, 3-chloropropyltrimethoxysilane, 3 methacryloyloxypropyltrimethoxysilane, 3-aminopropyltrimethoxysilane, 3 glycidoxypropyltrimethoxysilane, and 3-(triethoxysilyl)propylisocyanate. The intramolecular energy transfer process between lanthanide ions and the molecular bridges took place within these molecular-based hybrids and especially the quantum efficiency of europium hybrids were determined, suggesting that the hybrid material systems derived from different molecular bridges present different luminescence efficiencies. PMID- 18510294 TI - Fluorescent boron bis(phenolate) with association response to chloride and dissociation response to fluoride. AB - Addition of chloride ions to boron bis(phenolate) 5 in dichloromethane solution produces a selective fluorescence decrease. The fluorescence change is believed to be caused by associative hydrogen bonding between the chloride ion and two boronic acid groups. While addition of fluoride ions to bis(phenolate) 5 generates a purple colorimetric response, the colorimetric response is caused by fluoride induced B-O bond cleavage and air oxidation of the phenolate anion formed by this dissociation. PMID- 18510295 TI - On the residence time for water in a solute hydration shell: application to aqueous halide solutions. AB - We investigate several different methods to determine the water residence time next to a solute from molecular dynamics simulations. The popular computational prescription due to Impey et al. ( J. Phys. Chem. 1983, 87, 5071-5083) is shown to be extremely sensitive to the t* tolerance time value (designed to account for barrier recrossing effects), and we evidence through a kinetic analysis that the conventionally employed t* = 2 ps value can yield seriously overestimated residence times for low barrier exchanges. We suggest an alternate, robust determination based on the stable states picture (SSP) of chemical reactions ( J. Chem. Phys. 1980, 73, 2700-2714) where recrossing is naturally discarded. This is illustrated by calculation of the water residence time next to a water molecule and next to a chloride ion, using both nonpolarizable and polarizable force fields. The SSP results are in good agreement with the residence times estimated by a separate kinetic analysis, and differ noticeably from those calculated in the conventional fashion mentioned above. PMID- 18510296 TI - Fluorescence spectroscopy and amplified spontaneous emission (ASE) of phenylimidazoles: predicted vibronic coupling along the excited-state intramolecular proton transfer in 2-(2'-hydroxyphenyl)imidazoles. AB - Methylation at the 1N position of 2-phenylimidazole provides the shortest wavelength for a liquid-state laser dye reported to date; that is, the 1-methyl-2 phenylimidazole molecule in cyclohexane solution yields amplified spontaneous emission (ASE) with a peak wavelength at 314.5 nm and a constant laser gain value of 5 cm(-1) from 310 to 317 nm. Methyl substitution in this case favors the appearance of laser action (owing to a torsion-vibrational mechanism) in cyclohexane as compared with the nonmethylated species which does not exhibit ASE in this solvent. The 2-(2'-hydroxyphenyl)imidazole molecules give rise to ASE with high gain values (ca. 9 cm(-1)) at 450 and 466 nm. The mechanism of population inversion is understood in terms of a vibronic coupling between the hydroxyl stretching motion and the torsional vibration of the phenyl and imidazole rings. The proton-transfer spectroscopy of 2-(2' hydroxyphenyl)imidazoles is studied in dioxane, cyclohexane, dimethyl sulfoxide, methanol, and water. The greater the acidity of the solvent the greater the disruption of the intramolecular hydrogen bond; solvent acidity is the main parameter which favors formation of the open-form species in the ground electronic state. Methyl substitution at the 1N position favors formation of the open species for 2-hydroxyphenylimidazoles in the ground electronic state, which decreases their own capacity to undergo ASE. Low-temperature absorption spectroscopy confirms aggregation processes for 2-(2'-hydroxyphenyl)imidazoles in solution. In accordance with X-ray analyses in the solid phase, these molecules form associations through intermolecular chains of the type N-H...O or O-H...N. PMID- 18510297 TI - DFT study on isomerization and decomposition of cuprous dialkyldithiophosphate and its reaction with alkylperoxy radical. AB - The cuprous dialkyldithiophosphate [(RO)2PS2Cu, CuDDP] as an antioxidant has been industrially used in lubricating oil. In this paper, for the first time, a computational study has been carried out for CuDDP. The scaled hypersphere search method has been used to explore the isomerization and decomposition pathways of (HO)2PS2Cu, a model compound of CuDDP, and its reaction with CH3OO* radical. The calculations were performed at the B3LYP level of theory. The results show that the most stable structure of (HO)2PS2Cu has pseudo-C2v symmetry and a four membered ring constructed by P, Cu, and two S atoms. The bond-rearrangement isomerization leading to a (H)O-bridging structure is kinetically feasible. Three dissociation channels have been found for (HO)2PS2Cu, which require high energy (>60 kcal/mol) under the investigated condition. The reaction of (HO)2PS2Cu with CH3OO* includes bond-rearrangement isomerization and the decomposition of CH3OO* moiety. The (HO)2PS2Cu-assisted CH3OO* decomposition occurs via its O-O bond cleavage or the C-O bond dissociation. The former decomposition manner has been computed to be preferable over the latter at low temperature, but calculations suggested for the latter decomposition manner at higher temperature. Such a decomposition reaction, which is endothermic but possible, may be related to the antioxidation process of CuDDP. PMID- 18510298 TI - Mass spectrometry and photoelectron spectroscopy of o-, m-, and p-terphenyl cluster anions: the effect of molecular shape on molecular assembly and ion core character. AB - Mass spectrometry and photoelectron spectroscopy of o-, m-, and p-terphenyl cluster anions, (o-TP)n(-) (n = 2-100), (m-TP)n(-) (n = 2-100), and (p-TP)n(-) (n = 1-100), respectively, are conducted to investigate the effect of molecular shape on the molecular aggregation form and the resultant ion core character of the clusters. For (o-TP)n(-) and (m-TP)n(-), neither magic numbers nor discernible isomers are observed throughout the size range. Furthermore, their vertical detachment energies (VDEs) increase up to large n and depend linearly on n(-1/3), implying that they possess a three-dimensional (3D), highly reorganized structure encompassing a monomeric anion core. For (p-TP)n(-), in contrast, prominent magic numbers of n = 5, 7, 10, 12, and 14 are observed, and the VDEs show pronounced irregular shifts below n = 10, while they remain constant above n = 14 (isomer A). These results can be rationalized with two-dimensional (2D) orderings of p-TP molecules and different types of 2D shell closure at n = 7 and 14, the monomeric and multimeric anion core, respectively. Above n = 16, the new feature (isomer B) starts to appear at the higher binding side of isomer A, and it becomes dominant with n, while isomer A gradually disappears for larger sizes. In contrast to isomer A, the VDEs of isomer B continuously increase with the cluster size. This characteristic size evolution suggests that the transition to modified 2D aggregation forms from 2D ones occurs at around n = 20. PMID- 18510299 TI - Photophysics of (1-butyl-4-(1H-inden-1-ylidene)-1,4-dihydropyridine (BIDP): an experimental test for conical intersections. AB - Fluorescence experiments on (1-butyl-4-(1H-inden-1-ylidene)-1,4-dihydropyridine (BIDP) are reported in liquid and glassy solutions. The data indicate a fast decay in the fluid nonpolar, nonprotic solutions (decay times approximately 10( 12) s) and rapid but considerably slower decay in polar ones. In frozen solutions (polar and nonpolar), the fluorescence quantum yield is much higher (near 0.5 and around 0.1 in polar and nonpolar glasses, respectively). The rapid nonradiative transitions in fluid solutions are assigned to internal conversion in both solvent classes, as intersystem crossing is much slower and no net reaction is observed. These results are in agreement with predictions made for the closely related (in terms of electronic structure) but simpler molecule cyclopentadienyl 1,4-dihydropyridine (CPDHP) for which an S1/S0 conical intersection was recently proposed [Int. J. Quant. Chem. 2005, 102, 961]. The crossing of the two lowest singlet states is calculated to vanish in polar solvents such as methyl cyanide, leading to longer lifetime of S1 of CPDHP. As BIDP has a very similar electronic structure, the model predicts a corresponding change in this larger molecule. The strong fluorescence observed in the glassy environments is rationalized by the hindering of the internal torsion required to reach the geometry of the conical intersection. PMID- 18510300 TI - On the photophysics of polyenes. 1. Bathochromic shifts in their 1Ag --> 1Bu electronic transitions caused by the polarizability of the medium. AB - As shown in this study, the solvatochromic behavior of polyenes depends exclusively on the polarizability of the medium and, even more interestingly, their solvatochromism increases markedly with increasing length of the polyene chain. By virtue of the electronic nature of the interaction of polyenes with the medium, their solvatochromic response to a polarizability change is instantaneous, making these compounds extremely effective polarizability probes for molecular environments. The extreme sensitivity of polyenes to the polarizability of their environment is consistent with the fact that changes in molecular architecture such as those occurring in photosynthetic systems can give rise to polarizability gradients resulting in red shifts in the 1Ag --> 1Bu transition, thereby opening up new channels directing the energy transfer involved to energy trapping sites in such systems. PMID- 18510301 TI - Time-dependent density functional theory as a tool for isomer assignments of hydrogen-bonded solute.solvent clusters. AB - Can isomer structures of hydrogen-bonded solute x solvent clusters be assigned by correlating gas-phase experimental S0 <--> S1 transitions with vertical or adiabatic excitation energies calculated by time-dependent density functional theory (TD-DFT)? We study this question for 7-hydroxyquinoline (7HQ), for which an experimental database of 19 complexes and clusters is available. The main advantage of the adiabatic TD-B3LYP S0 <--> S1 excitations is the small absolute error compared to experiment, while for the calculated vertical excitations, the average offset is +1810 cm(-1). However, the empirically adjusted vertical excitations correlate more closely with the experimental transition energies, with a standard deviation of sigma = 72 cm(-1). For the analogous correlation with calculated adiabatic TD-DFT excitations, the standard deviation is sigma = 157 cm(-1). The vertical and adiabatic TD-DFT correlation methods are applied for the identification of isomers of the 7-hydroxyquinoline.(MeOH) n , n = 1-3 clusters [Matsumoto, Y.; Ebata, T.; Mikami, N. J. Phys. Chem. B 2002, 106, 5591]. These confirm that the vertical TD-DFT/experimental correlation yields more effective isomer assignments. PMID- 18510302 TI - Separation and complete analyses of the overlapped and unresolved 1H NMR spectra of enantiomers by spin selected correlation experiments. AB - NMR spectroscopic discrimination of optical enantiomers is most often carried out using (2)H and (13)C spectra of chiral molecules aligned in a chiral liquid crystalline solvent. The use of proton NMR for such a purpose is severely hindered due to the spectral complexity and the significant loss of resolution arising from numerous short- and long-distance couplings and the indistinguishable overlap of spectra from both R and S enantiomers. The determination of all the spectral parameters by the analyses of such intricate NMR spectra poses challenges, such as, unraveling of the resonances for each enantiomer, spectral resolution, and simplification of the multiplet pattern. The present study exploits the spin state selection achieved by the two-dimensional (1)H NMR correlation of selectively excited isolated coupled spins (Soft-COSY) of the molecules to overcome these problems. The experiment provides the relative signs and magnitudes of all of the proton-proton couplings, which are otherwise not possible to determine from the broad and featureless one-dimensional (1)H spectra. The utilization of the method for quantification of enantiomeric excess has been demonstrated. The studies on different chiral molecules, each having a chiral center, whose spectral complexity increases with the increasing number of interacting spins, and the advantages and limitations of the method over SERF and DQ-SERF experiments have been reported in this work. PMID- 18510303 TI - Matrix isolation infrared spectroscopic and theoretical study of the hydrolysis of boron dioxide in solid argon. AB - The reaction of boron dioxide with water molecule has been studied using matrix isolation infrared spectroscopy. The boron dioxide molecules produced by codeposition of laser-evaporated boron atoms with dioxygen react spontaneously with water molecules to form OB(OH)2, which is characterized to have a doublet ground-state with two OH groups in the cis-trans form. Isotopic substitution results indicate that the hydrolysis process proceeds via a concerted two hydrogen atom transfer mechanism. The cis-trans-OB(OH)2 molecule is photosensitive; it decomposes to the OH x OB(OH) complex upon broadband UV visible irradiation. The OH x OB(OH) complex is determined to have a (2)A'' ground-state with a bent C(s) symmetry, in which the terminal oxygen atom of the OB(OH) fragment is hydrogen bonded with the hydroxyl radical. The OH x OB(OH) complex recombines to the cis-trans-OB(OH)2 molecule upon sample annealing. PMID- 18510304 TI - Enthalpies of formation of hydrocarbons by hydrogen atom counting. Theoretical implications. AB - Standard enthalpies of formation at 298 K of unstrained alkanes, alkenes, alkynes, and alkylbenzenes can be expressed as a simple sum in which each term consists of the number of hydrogen atoms n of one of eight different types (n1 n8) multiplied by an associated coefficient (c1-c8) derived from the known enthalpy of formation of a typical molecule. Alkylbenzenes require one additive constant for each benzene ring, accounting for a possible ninth term in the sum. Terms are not needed to account for repulsive or attractive 1,3 interactions, hyperconjugation, or for protobranching, rendering them irrelevant. Conjugated eneynes and diynes show thermodynamic stabilizations much smaller than that observed for 1,3-butadiene, bringing into question the usual explanation for the thermodynamic stabilization of conjugated multiple bonds (p orbital overlap, pi electron delocalization, etc.). PMID- 18510305 TI - High-stability hydrogenated silicon-carbon clusters: a full study of Si2C2H2 in comparison to Si2C2, C2B2H4, and other similar species. AB - The structural and electronic characteristics of the Si2C2H2 and Si2C2 clusters are studied by ab initio calculations based on coupled cluster and density functional theory using the hybrid B3LYP functional. In addition, similar species, such as SiC2H2 and Si3C2H2, are also studied for comparison. It is illustrated that the lowest energy structures of all three hydrogenated clusters, which have the general form Si(n)(CH)2, n = 1, 2, 3, are fully analogous to the structures of the corresponding organometallic isovalent carboranes. The most stable structure of Si2C2H2 is obtained by attaching two hydrogens onto the carbon atoms of a higher energy (+1.5 eV) planar trapezoidal structure of Si2C2, followed by geometry optimization which leads to puckering of the planar structure. Furthermore, it is demonstrated that Si2C2H2 and the other two "similar" hydrogenated clusters are much more stable than the corresponding bare nonhydrogenated clusters. Comparison of Si2C2H2 and C2B2H4 shows that their structural and bonding similarity includes also nuclear rearrangement similarity. The two species are isomerizable with an energy difference between their lowest energy puckered 1,2- and 1,3-isomers of about +/-0.3 eV. It is suggested that SiC2H2, Si2C2H2, and Si3C2H2 are special cases of a larger class of stable clusters. It is speculated on the basis of the calculated infrared spectrum that Si2C2H2 and perhaps other members of this class of clusters could be found in appreciable abundance in interstellar space. PMID- 18510306 TI - Docking to RNA via root-mean-square-deviation-driven energy minimization with flexible ligands and flexible targets. AB - Structure-based drug design is now well-established for proteins as a key first step in the lengthy process of developing new drugs. In many ways, RNA may be a better target to treat disease than a protein because it is upstream in the translation pathway, so inhibiting a single mRNA molecule could prevent the production of thousands of protein gene products. Virtual screening is often the starting point for structure-based drug design. However, computational docking of a small molecule to RNA seems to be more challenging than that to protein due to the higher intrinsic flexibility and highly charged structure of RNA. Previous attempts at docking to RNA showed the need for a new approach. We present here a novel algorithm using molecular simulation techniques to account for both nucleic acid and ligand flexibility. In this approach, with both the ligand and the receptor permitted some flexibility, they can bind one another via an induced fit, as the flexible ligand probes the surface of the receptor. A possible ligand can explore a low-energy path at the surface of the receptor by carrying out energy minimization with root-mean-square-distance constraints. Our procedure was tested on 57 RNA complexes (33 crystal and 24 NMR structures); this is the largest data set to date to reproduce experimental RNA binding poses. With our procedure, the lowest-energy conformations reproduced the experimental binding poses within an atomic root-mean-square deviation of 2.5 A for 74% of tested complexes. PMID- 18510307 TI - Radical-ligand-derived C-N coupling, Ga(III)-radical vs low-spin Co(III)-radical reactivity. AB - The redox-active ligand 2-(3,5-dimethoxyanilino)-4,6-di- tert-butylphenol, H 2L (OCH3), results in, as expected, a trisradical complex with a low-spin Co(III) center, [Co (III)(L (OCH3) (*)) 3] ( 1), whereas the Ga(III) center yields a coordinated new hexadentate monoradical ligand, [Ga (III)L (*) 1] ( 2), presumably due to the ligand-derived redox activity involving C-H activation. PMID- 18510308 TI - Tin-based organo-Zintl ions: alkylation and alkenylation of Sn9(4-). AB - Reactions of nine-atom deltahedral clusters (Zintl ions) of tin, Sn 9 (4-), with alkyl chlorides, RCl (R = (t) Bu, (n) Bu, (s) Bu), and alkynes (Me3Si-C[triple bond]C-SiMe3, Ph-C[triple bond]CH) yielded the corresponding alkylated and alkenylated clusters [Sn 9-R] (3-). The triple bonds of the alkynes are hydrogenated to double bonds in the process. These are the first tin-based organo Zintl ions, that is Zintl ions of tin that were subsequently functionalized with organic groups. They are analogous to the recently reported germanium-based derivatives. The (t) Bu-, vinyl-, and styrene-functionalized clusters [Sn 9- (t) Bu] (3-), [Sn 9-CH=CH 2] (3-), and [Sn 9-CH=CH-Ph] (3-), respectively, were structurally characterized in the solid state with [K(2,2,2-crypt)] (+) countercations and in solution by electrospray mass spectrometry. Crystal data: [K(2,2,2-crypt)] 3[Sn 9- (t) Bu].2py, triclinic, P1, a = 14.4259(3), b = 16.2725(4), and c = 22.5593(5) A, alpha = 86.092(1), beta = 78.952(1), and gamma = 65.114(1) degrees , V = 4714.48(7) A (3), Z = 2; [K(2,2,2-crypt)] 3[Sn 9-CH=CH 2].2py, triclinic, P-1, a = 15.6988(3), b = 17.4195(4), and c = 17.4432(4) A, alpha = 86.299(1), beta = 81.566(1), and gamma = 85.349(1) degrees , V = 4696.27(18) A (3), Z = 2; [K(2,2,2-crypt)] 3[Sn 9-CH=CH-Ph].tol.0.75py, monoclinic, C2/c, a = 38.5883(9), b = 23.3893(5), and c = 25.0192(5) A, beta = 120.269(1) degrees , V = 19502.6(7) A (3), Z = 8. PMID- 18510309 TI - General synthesis of (salen)ruthenium(III) complexes via N...N coupling of (salen)ruthenium(VI) nitrides. AB - Reaction of [Ru (VI)(N)(L (1))(MeOH)] (+) (L (1) = N, N'-bis(salicylidene)- o cyclohexylenediamine dianion) with excess pyridine in CH 3CN produces [Ru (III)(L (1))(py) 2] (+) and N 2. The proposed mechanism involves initial equilibrium formation of [Ru (VI)(N)(L (1))(py)] (+), which undergoes rapid N...N coupling to produce [(py)(L (1))Ru (III) N N-Ru (III)(L (1))(py)] (2+); this is followed by pyridine substituion to give the final product. This ligand-induced N...N coupling of Ru (VI)N is utilized in the preparation of a series of new ruthenium(III) salen complexes, [Ru (III)(L)(X) 2] (+/-) (L = salen ligand; X = H 2O, 1-MeIm, py, Me 2SO, PhNH 2, ( t )BuNH 2, Cl (-) or CN (-)). The structures of [Ru (III)(L (1))(NH 2Ph) 2](PF 6) ( 6), K[Ru (III)(L (1))(CN) 2] ( 9), [Ru (III)(L (2))(NCCH 3) 2][Au (I)(CN) 2] ( 11) (L (2) = N, N'-bis(salicylidene)- o phenylenediamine dianion) and [N ( n )Bu 4][Ru (III)(L (3))Cl 2] ( 12) (L (3) = N, N'-bis(salicylidene)ethylenediamine dianion) have been determined by X-ray crystallography. PMID- 18510310 TI - Effect of chelate dynamics on water exchange reactions of paramagnetic aminopolycarboxylate complexes. AB - Because of our interest in evaluating a possible relationship between complex dynamics and water exchange reactivity, we performed (1)H NMR studies on the paramagnetic aminopolycarboxylate complexes Fe (II)-TMDTA and Fe (II)-CyDTA and their diamagnetic analogues Zn (II)-TMDTA and Zn (II)-CyDTA. Whereas a fast Delta Lambda isomerization was observed for the TMDTA species, no acetate scrambling between in-plane and out-of-plane positions is accessible for any of the CyDTA complexes because the rigid ligand backbone prevents any configurational changes in the chelate system. In variable-temperature (1)H NMR studies, no evidence of spectral coalescence due to nitrogen inversion was found for any of the complexes in the available temperature range. The TMDTA complexes exhibit the known solution behavior of EDTA, whereas the CyDTA complexes adopt static solution structures. Comparing the exchange kinetics of flexible EDTA-type complexes and static CyDTA complexes appears to be a suitable method for evaluating the effect of ligand dynamics on the overall reactivity. In order to assess information concerning the rates and mechanism of water exchange, we performed variable temperature and -pressure (17)O NMR studies of Ni (II)-CyDTA, Fe (II)-CyDTA, and Mn (II)-CyDTA. For Ni (II)-CyDTA, no significant effects on line widths or chemical shifts were apparent, indicating either the absence of any chemical exchange or the existence of a very small amount of the water-coordinated complex in solution. For [Fe (II)(CyDTA)(H 2O)] (2-) and [Mn (II)(CyDTA)(H 2O)] (2-), exchange rate constant values of (1.1 +/- 0.3) x 10 (6) and (1.4 +/- 0.2) x 10 (8) s (-1), respectively, at 298 K were determined from fits to resonance-shift and line-broadening data. A relationship between chelate dynamics and reactivity seems to be operative, since the CyDTA complexes exhibited significantly slower reactions than their EDTA counterparts. The variable-pressure (17)O NMR measurements for [Mn (II)(CyDTA)(H 2O)] (2-) yielded an activation volume of +9.4 +/- 0.9 cm (3) mol (-1). The mechanism is reliably assigned as a dissociative interchange (I d) mechanism with a pronounced dissociation of the leaving water molecule in the transition state. In the case of [Fe (II)(CyDTA)(H 2O)] (2-), no suitable experimental conditions for variable-pressure measurements were accessible. PMID- 18510311 TI - Sarcoplasmic reticulum calcium ATPase is inhibited by organic vanadium coordination compounds: pyridine-2,6-dicarboxylatodioxovanadium(V), BMOV, and an amavadine analogue. AB - The general affinity of the sarcoplasmic reticulum (SR) Ca (2+)-ATPase was examined for three different classes of vanadium coordination complexes including a vanadium(V) compound, pyridine-2,6-dicarboxylatodioxovanadium(V) (PDC-V(V)), and two vanadium(IV) compounds, bis(maltolato)oxovanadium(IV) (BMOV), and an analogue of amavadine, bis( N-hydroxylamidoiminodiacetato)vanadium(IV) (HAIDA V(IV)). The ability of vanadate to act either as a phosphate analogue or as a transition-state analogue with enzymes' catalysis phosphoryl group transfer suggests that vanadium coordination compounds may reveal mechanistic preferences in these classes of enzymes. Two of these compounds investigated, PDC-V(V) and BMOV, were hydrolytically and oxidatively reactive at neutral pH, and one, HAIDA V(IV), does not hydrolyze, oxidize, or otherwise decompose to a measurable extent during the enzyme assay. The SR Ca (2+)-ATPase was inhibited by all three of these complexes. The relative order of inhibition was PDC-V(V) > BMOV > vanadate > HAIDA-V(IV), and the IC 50 values were 25, 40, 80, and 325 microM, respectively. Because the observed inhibition is more potent for PDC-V(V) and BMOV than that of oxovanadates, the inhibition cannot be explained by oxovanadate formation during enzyme assays. Furthermore, the hydrolytically and redox stable amavadine analogue HAIDA-V(IV) inhibited the Ca (2+)-ATPase less than oxovanadates. To gauge the importance of the lipid environment, studies of oxidized BMOV in microemulsions were performed and showed that this system remained in the aqueous pool even though PDC-V(V) is able to penetrate lipid interfaces. These findings suggest that the hydrolytic properties of these complexes may be important in the inhibition of the calcium pump. Our results show that two simple coordination complexes with known insulin enhancing effects can invoke a response in calcium homeostasis and the regulation of muscle contraction through the SR Ca (2+)-ATPase. PMID- 18510312 TI - D(+)-pi-A(-) charge-transfer molecules based on tricyanoquinodimethane and diphosphine metal complexes. AB - A new family of D (+)-pi-A (-) chromophores in which the donor group is an organometallic complex and the acceptor group a tricyanoquinodimethane moiety has been synthesized by the reaction of diphosphinomethanide transition-metal complexes and 7,7',8,8'-tetracyanoquinodimethane. PMID- 18510313 TI - Cell-based fluorescent indicator to visualize brain-derived neurotrophic factor secreted from living neurons. AB - Brain-derived neurotrophic factor (BDNF) is a polypeptide that is secreted from neurons. Although there is mounting evidence that BDNF regulates neuronal development and synaptic plasticity, BDNF secretion has remained unclear due to lack of appropriate methods for the analysis of its dynamics. To visualize BDNF secretion from neurons, here we have developed a cell-based fluorescent indicator for BDNF. We showed that the present cell-based fluorescent indicator, named "Bescell", has high selectivity to BDNF and detects picomolar concentrations of BDNF (detection limit of 60 pM). Bescell has visualized endogenous BDNF secreted from hippocampal neurons. It thus provides a powerful tool for the analysis of BDNF secretion from living neurons. PMID- 18510314 TI - Evaluation of the carbene hydride mechanism in the carbon-carbon bond formation process of alkane metathesis through a DFT study. AB - Olefin metathesis on a silica supported tantalumhydridocarbene complex, the key carbon-carbon making process in alkane metathesis, requires a large number of elementary steps in contrast to the known olefin metathesis pathway, which corresponds to successive [2 + 2]-cycloaddition and cycloreversion steps. The direct pathway is forbidden because it requires the formation of a high energy reaction intermediates, an olefin adduct of trigonal bipyramid (TBP) geometry, where the carbene is trans to an hydride ligand. Extra low-energy steps are therefore necessary to connect the reactants to products, the key being a turnstile interconversion at the metallacyclobutane intermediates. PMID- 18510315 TI - Double-stranded helical polymers consisting of complementary homopolymers. AB - Two complementary homopolymers of chiral amidines and achiral carboxylic acids with m-terphenyl-based backbones were synthesized by the copolymerization of a p diiodobenzene derivative with the diethynyl monomers bearing a chiral amidine group and a carboxyl group using the Sonogashira reaction, respectively. Upon mixing in THF, the homopolymer strands assembled into a preferred-handed double helix through interstrand amidinium-carboxylate salt bridges, as evidenced by its absorption, circular dichroism, and IR spectra. In contrast, when mixed in less polar solvents, such as chloroform, the complementary strands kinetically formed an interpolymer complex with an imperfect double helical structure containing a randomly hybridized cross-linked structure, probably because of strong salt bridge formations. This primary complex was rearranged into the fully double helical structure by treatment with a strong acid followed by neutralization with an amine. High-resolution atomic force microscopy revealed the double-stranded helical structure and enabled the determination of the helical sense. PMID- 18510317 TI - Toward reliable gold nanoparticle patterning on self-assembled DNA nanoscaffold. AB - Assembly of gold nanoparticles (AuNP) into designer architectures with reliablity is important for nanophotonics and nanoelectronics applications. Toward this goal we present a new strategy to prepare AuNPs monofunctionalized with lipoic acid modified DNA oligos. This strategy offers increased bonding strength between DNA oligos and AuNP surface. These conjugates are further selectively mixed with other DNA strands and assembled into fixed sized DNA nanostructures carring a discrete number of AuNPs at desired positions. Atomic force microscopy imaging reveals a dramatically improved yield of the AuNPs on DNA tile structure compared to the ensembles using monothiolate AuNP-DNA conjugates. PMID- 18510318 TI - One-step multicomponent encapsulation by compound-fluidic electrospray. AB - Fabrication of sophisticated or smart materials often needs controlled integrating multiple components into a single capsule. Most of conventional microencapsulation strategies merely envelop one content into a shell every time. We report a compound-fluidic electrospray method could one-step enclose multiple components into a single microcapsule without contact. The as-prepared microcapsules have multiple compartments inside, in each of which different content can be addressably loaded. This approach gives flexibility for generating diverse microcapsules that could one-step integrate different active components in microscopic domain free of contact, which may find potential applications in multicomponent drug delivery, microreactors and others. PMID- 18510319 TI - Design of an N-methylated peptide inhibitor of alpha-synuclein aggregation guided by solid-state NMR. AB - Many neurodegenerative diseases are associated with the aggregation of misfolded proteins into amyloid oligomers or fibrils that are deposited as pathological lesions within areas of the brain. An attractive therapeutic strategy for preventing or ameliorating amyloid formation is to identify agents that inhibit the onset or propagation of protein aggregation. Here we demonstrate how solid state nuclear magnetic resonance (ssNMR) may be used to identify key residues within amyloidogenic protein sequences that may be targeted to inhibit the aggregation of the host protein. For alpha-synuclein, the major protein component of Lewy bodies associated with Parkinson's disease, we have used a combination of ssNMR and biochemical data to identify the key region for self-aggregation of the protein as residues 77-82 (VAQKTV). We used our new structural information to design a peptide derived from residues 77 to 82 of alpha-synuclein with an N methyl group at the C-terminal residue, which was able to disrupt the aggregation of alpha-synuclein. Thus, we have shown how structural data obtained from ssNMR can guide the design of modified peptides for use as amyloid inhibitors, as a primary step toward developing therapeutic compounds for prevention and/or treatment of amyloid diseases. PMID- 18510320 TI - Asymmetric aza-Henry reaction under phase transfer catalysis: an experimental and theoretical study. AB - An efficient catalytic asymmetric aza-Henry reaction under phase transfer conditions is presented. The method is based on the reaction of the respective nitroalkane with alpha-amido sulfones effected by CsOH x H2O base in toluene as solvent and in the presence of cinchone-derived ammonium catalysts. This direct aza-Henry reaction presents as interesting features its validity for both nonenolizable and enolizable aldehyde-derived azomethines and the tolerance of nitroalkanes, other than nitromethane, for the production of beta-nitroamines. The synthetic value of the methodology described is demonstrated by providing (a) a direct route for the asymmetric synthesis of differently substituted 1,2 diamines and (b) a new asymmetric synthesis of gamma-amino alpha,beta-unsaturated esters through a catalytic, highly enantioselective formal addition of functionalized alkenyl groups to azomethines. Finally, a preferred TS that nicely fits the observed enantioselectivity has been identified. Most remarkable, an unusual hydrogen bond pattern for the catalyst-nitrocompound-imine complex is predicted, where the catalyst OH group interacts with the NO2 group of the nitrocompound. PMID- 18510321 TI - Synthesis and reactivity studies of iminoboryl complexes. AB - A range of new iminoborylcomplexes of the type [L(n)M-B[triple bond]N-R], which are isoelectronic with sigma-alkynyl complexes [L(n)M-C[triple bond]C-R], was obtained by systematically varying the metal M, the coligands L, and the nitrogen bound substituent R. Selected examples include, for example, trans [(Cy3P)2(Br)Pt(B[triple bond]N iBu)], which is characterized by a sterically less demanding N-R group or the unprecedented rhodium species cis,mer [(Br)2(Me3P)3Rh(B[triple bond]NSiMe3)]. All compounds were fully characterized in solution by multinuclear NMR spectroscopy and, where appropriate, in the solid state by X-ray crystallography. Subsequent reactivity studies revealed that particularly the combination of smaller N-R groups with Pt-B linkages of increased stability opens up opportunities for novel reactivity patterns of this class of compounds. Within the scope of these study, we inter alia succeeded in synthesizing the unusual bridged boryl species 1,4-trans-[{(Cy3P)2(Br)Pt(B{NH iBu}NH)}2C6H4] and a complex bearing both an acetylide ligand and an iminoboryl ligand, respectively. PMID- 18510322 TI - Functional mimicry of carboxypeptidase A by a combination of transition state stabilization and a defined orientation of catalytic moieties in molecularly imprinted polymers. AB - An artificial model for the natural enzyme carboxypeptidase A has been constructed by molecular imprinting in synthetic polymers. The tetrahedral transition state analogues (TSAs 4 and 5) for the carbonate hydrolysis have been designed as templates to allow incorporation of the main catalytic elements, an amidinium group and a Zn(2+) or Cu(2+) center, in a defined orientation in the transition state imprinted active site. The complexation of the functional monomer and the template in presence of Cu(2+) through stoichiometric noncovalent interaction was established on the basis of (1)H NMR studies and potentiometric titration. The Cu(2+) center was introduced into the imprinted cavity during polymerization or by substitution of Zn(2+) in Zn(2+) imprinted polymers. The direct introduction displayed obvious advantages in promoting catalytic efficiency. With substrates exhibiting a very similar structure to the template, an extraordinarily high enhancement of the rate of catalyzed to uncatalyzed reaction (k(cat)/k(uncat)) of 10(5)-fold was observed. If two amidinium moieties are introduced in proximity to one Cu(2+) center in the imprinted cavity by complexation of the functional monomer 3 with the template 5, the imprinted catalysts exhibited even higher activities and efficiencies for the carbonate hydrolysis with k(cat)/k(uncat) as high as 410,000. These are by far the highest values obtained for molecularly imprinted catalysts, and they are also considerably higher compared to catalytic antibodies. Our kinetic studies and competitive inhibition experiments with the TSA template showed a clear indication of a very efficient imprinting procedure. In addition, this demonstrates the important role of the transition state stabilization during the catalysis of this reaction. PMID- 18510323 TI - Beyond Aresta's complex: Ni- and Pd-catalyzed organozinc coupling with CO2. AB - Organozinc reagents are widely used in organic synthesis due primarily to their high functional group compatibility. However, under mild conditions, these organometallic reagents do not react directly with CO2, the ideal C1 source for organic synthesis. Herein, we report that both Ni and Pd complexes can catalyze the addition of organozinc reagents to CO2 under mild conditions (1 atm CO2, 0 degrees C). This transformation represents an important extension of the Negishi cross-coupling and is believed to occur via a novel mechanism involving Aresta's complex and/or its corresponding Pd analogue. PMID- 18510324 TI - Self-assembly of surfactants and polymorphic transition in nanotubes. AB - We study self-assembly and polymorphic transitions of surfactant molecules in water within a nanotube and the effect of water-nanotube interactions on the self assembly morphologies. We present a simulation evidence of a cornucopia of polymorphic structures of surfactant assemblies--many of which have not been observed in bulk solutions--through adjusting the water-nanotube chemical interactions which range from hydrophilic to hydroneutral and to hydrophobic. The ability to control the morphologies of surfactant assemblies within nanoscale confinement can be used for patterning the interior surface of nanochannels for application in nanofluidics and nanomedical devices. PMID- 18510325 TI - Temperature controlled reversible change of the coordination modes of the highly symmetrical multitopic ligand to construct coordination assemblies: experimental and theoretical studies. AB - By fine-tuning the reaction temperature to 5, 15, 30, and 50 degrees C, respectively, four different complexes, [Cd(HC4O4)2(H2O)4] (1), [Cd4(C4O4)4(H2O)16].(H2O)2 (2), [Cd(C4O4)(H2O)2] (3), and [Cd(C4O4)(H2O)2] (4), were formed successfully from the identical initial reaction mixture. Moreover, an unprecedented reversible interconversion among the four complexes at the corresponding reaction temperatures mediated by the mother liquor was observed. PMID- 18510326 TI - A supramolecular receptor of diatomic molecules (O2, CO, NO) in aqueous solution. AB - A per-O-methylated beta-cyclodextrin dimer, Py2CD, was conveniently prepared via two steps: the Williamson reaction of 3,5-bis(bromomethyl)pyridine and beta cyclodextrin (beta-CD) yielding 2A,2'A-O-[3,5-pyridinediylbis(methylene)bis-beta cyclodextrin (bisCD) followed by the O-methylation of all the hydroxy groups of the bisCD. Py2CD formed a very stable 1:1 complex (Fe(III)PCD) with [5,10,15,20 tetrakis(p-sulfonatophenyl)porphinato]iron(III) (Fe(III)TPPS) in aqueous solution. Fe(III)PCD was reduced with Na2S2O4 to afford the Fe (II)TPPS/Py2CD complex (Fe(II)PCD). Dioxygen was bound to Fe(II)PCD, the P(1/2)(O2) values being 42.4 +/- 1.6 and 176 +/- 3 Torr at 3 and 25 degrees C, respectively. The k(on)(O2) and k(off)(O2) values for the dioxygen binding were determined to be 1.3 x 10(7) M(-1) s(-1) and 3.8 x 10(3) s(-1), respectively, at 25 degrees C. Although the dioxygen adduct was not very stable (K(O2) = k(on)(O2)/k(off)(O2) = 3.4 x 10(3) M(-1)), no autoxidation of the dioxygen adduct of Fe(II)PCD to Fe(III)PCD was observed. These results suggest that the encapsulation of Fe (II)TPPS by Py2CD strictly inhibits not only the extrusion of dioxygen from the cyclodextrin cage but also the penetration of a water molecule into the cage. The carbon monoxide affinity of Fe(II)PCD was much higher than the dioxygen affinity; the P(1/2)(CO), k(on)(CO), k(off)(CO), and K(CO) values being (1.6 +/- 0.2) x 10( 2) Torr, 2.4 x 10(6) M(-1) s(-1), 4.8 x 10(-2) s(-1), and 5.0 x 10(7) M(-1), respectively, at 25 degrees C. Fe(II)PCD also bound nitric oxide. The rate of the dissociation of NO from (NO)Fe(II)PCD ((5.58 +/- 0.42) x 10(-5) s(-1)) was in good agreement with the maximum rate ((5.12 +/- 0.18) x 10(-5) s(-1)) of the oxidation of (NO)Fe(II)PCD to Fe(III)PCD and NO3(-), suggesting that the autoxidation of (NO)Fe(II)PCD proceeds through the ligand exchange between NO and O2 followed by the rapid reaction of (O2)Fe(II)PCD with released NO, affording Fe(II)PCD and the NO3(-) anion inside the cyclodextrin cage. PMID- 18510327 TI - Quantum size effect directed selective self-assembling of cobalt phthalocyanine on Pb(111) thin films. AB - Adsorption and self-assembly of cobalt phthalocyanine (CoPc) molecules on Pb(111) thin films with a thickness ranging from 10 atomic monolayers (ML) to 20 ML were investigated by using scanning tunneling microscopy and spectroscopy (STM/STS). Unprecedented thickness-selective oscillating adsorption and self-assembly behavior of the molecules on the films were observed. STS measurement reveals that this oscillatory behavior arises from quantum size effect. The strong quantum confinement of electron motion in the Pb films modulates the electronic density of states at the Fermi level (DOS(EF)), leading to preferential adsorption at thicknesses of higher DOS(EF). The work provides an unambiguous evidence for quantum modulation of surface reactivities of a metal thin film. PMID- 18510328 TI - A new five-membered ring forming process based on palladium(0)-catalyzed arylative cyclization of allenyl enones. AB - A palladium(0)/monophosphine catalyst promotes a novel arylative cyclization reaction of C1-, C2-, and C3-tethered allenyl enones with arylboronic acids to produce five-membered ring containing products. The regioselectivity of the process, associated with aryl group introduction into the allene moiety, depends on the length of the tether. This finding suggests that the cyclization reaction does not proceed through a carbopalladation pathway but rather via a route involving palladacycle-forming or "anti-Wacker"-type oxidative addition to the Pd(0) catalyst. PMID- 18510329 TI - Photogeneration and thermal generation of pentacene from soluble precursors for OTFT applications. AB - A CO adduct of pentacene with an unsymmetrical structure is synthesized; it is soluble and can be spin-coated into thin films. Pentacene is regenerated in near quantitative yield by either thermal or photoinduced elimination of CO. OTFT devices fabricated by this compound exhibit typical FET characteristics. PMID- 18510330 TI - Successive copper(I)-catalyzed cross-couplings in one pot: a novel and efficient starting point for synthesis of carbapenems. AB - An efficient approach for synthesizing a series of 2-sulfide carbapenems has been developed using two successive Cu(I)-catalyzed cross-couplings in a single pot. The method involves highly selective intramolecular coupling of lactam and dihaloalkene using 2,2'-bipyridine as a ligand, followed by intermolecular C-S formation in the presence of another ligand (1,10-phenanthroline, PPh 3) and mercaptan. PMID- 18510331 TI - Olefin metathesis catalyst: stabilization effect of backbone substitutions of N heterocyclic carbene. AB - Ruthenium olefin metathesis catalysts bearing an N-phenyl-substituted N heterocyclic carbene (NHC) ligand that are resistant to decomposition through C-H activation have been prepared and tested in ring closing metathesis (RCM), cross metathesis (CM), and ROMP reactions. The N, N'-diphenyl-substituted NHC complex proved to be one of the most efficient catalysts in RCM to form tetrasubstituted olefins. PMID- 18510332 TI - Synthesis of the macrocyclic core of (-)-pladienolide B. AB - An efficient synthesis of the macrocyclic core of (-)-pladienolide B is disclosed. The concise route relies on a chiral auxiliary-mediated asymmetric aldol addition and an osmium-catalyzed asymmetric dihydroxylation to install the three oxygenated stereocenters of the macrocycle. This purely reagent-controlled and flexible strategy sets the stage for future analogue syntheses and structure activity relationship plotting of the appealing anticancer lead structure pladienolide B. PMID- 18510333 TI - Diversity synthesis of complex pyridines yields a probe of a neurotrophic signaling pathway. AB - Recognizing the value of including complex pyridines in small-molecule screening collections, we developed a previously unexplored [2 + 2 + 2]-cycloaddition of silyl-tethered diynes with nitriles. The tether provides high regioselectivity, while the solvent THF allows catalytic CpCo(CO)(2) to be used without exogenous irradiation. One of the resulting bicyclic and monocyclic (desilylated) pyridines was identified as an inhibitor of neuregulin-induced neurite outgrowth (EC(50) = 0.30 microM) in a screen that probes a pathway likely to be involved in breast cancers and schizophrenia. PMID- 18510334 TI - Asymmetric synthesis of carboxylic acid derivatives having an all-carbon alpha quaternary center through Cu-catalyzed 1,4-addition of dialkylzinc reagents to 2 aryl acetate derivatives. AB - The asymmetric synthesis of carboxylic acid derivatives having an all-carbon alpha-quaternary center has been achieved via copper-catalyzed 1,4-addition of dialkylzinc reagents to aryl acetate derivatives in the presence of phosphoramidite ligand. High isolated yields and enantioselectivities were obtained. It was demonstrated that the Meldrum's acid and ester moieties present on the all-carbon quaternary center allow for a wide variety of subsequent transformations, leading to the expedient preparation of succinimides, succinate esters and succinic acids, gamma-butyrolactones, and beta-amino acid derivatives. PMID- 18510335 TI - Oxygen concentration affects volatile compound biosynthesis during virgin olive oil production. AB - The effect of O 2 concentration on oil volatile compounds synthesized during the process to obtain virgin olive oil (VOO) was established. The study was carried out either on the whole process or within the main steps (milling and malaxation) of this process with two olive cultivars, Picual and Arbequina, at two ripening stages. Data show that O 2 control during milling has a negative impact on VOO volatile synthesis. This effect seems to depend on cultivar and on the ripening stage in cultivar Picual. Because most VOO volatiles are synthesized during olive fruit crushing at the milling step, O 2 control during malaxation seems to affect just slightly the volatile synthesis. The highest effect was observed when control of O 2 concentration was performed over the whole process. In this case, the content of volatile compounds of oils obtained from both cultivars and ripening stages showed quite similar trends. PMID- 18510336 TI - Characterization of a benzyl alcohol dehydrogenase from Lactobacillus plantarum WCFS1. AB - Aroma is an important sensory parameter of food products. Lactic acid bacteria have enzymatic activities that could be important in the modification of food aroma. The complete genome sequence from Lactobacillus plantarum WCFS1 shows a gene (lp_3054) putatively encoding a protein with benzyl alcohol dehydrogenase activity. To confirm its enzymatic activity lp_3054 from this strain has been overexpressed and purified. Protein alignment indicated that lp_3054 is a member of the family of NAD(P)-dependent long-chain zinc-dependent alcohol dehydrogenases. In lp_3054 all of the residues involved in zinc and cofactor binding are conserved. It is also conserved the residue that determines the specificity of the dehydrogenase toward NAD (+) rather than NADP (+) and, therefore, L. plantarum benzyl alcohol dehydrogenase is less active in the presence of NADP (+) than in the presence of NAD (+). The purified enzyme exhibits optimal activity at pH 5.0 and 30 degrees C. The kinetic parameters K m and V max on benzyl alcohol as a substrate were, respectively, 0.23 mM and 204 mumol h (-1) mg (-1). Besides its activity toward benzyl alcohol, it showed activity against nerol, geraniol, phenethyl alcohol, cinnamyl alcohol, and coniferyl alcohol, all of which are volatile compounds involved in determining food aroma. The biochemical demonstration of a functional benzyl alcohol dehydrogenase activity in this lactic acid bacteria species should be considered when the influence of bacterial metabolism in the aroma of food products is determined. PMID- 18510337 TI - Effects of Triton X-100 nanoaggregates on dimerization and antioxidant activity of morin. AB - Dimerization and antioxidant activity of morin in the Triton X-100 micelles were studied by electronic absorption, ATR-FTIR spectra, cyclic voltammetric, DSC, freeze-fracture TEM, molecular modeling and ab initio quantum calculations. Morin can be solubilized in the Triton X-100 micelles and show selective dimerization in Triton X-100 micelles with different structures. In Triton X-100 spherical micelles, morin always exists in the form of dimer, and in Triton X-100 rodlike micelles, it is always in the form of monomer. The solubilization of morin dimer in Triton X-100 spherical micelles changes the micelle morphology from spherical to cubelike, and the size of the single micelle is also increased, while morin monomer links the Triton X-100 rodlike micelles and forms a kind of network micelle structure with the size of the "rod" unchanged. Solubilized and concentrated in Triton X-100 micelles, morin can protect human serum albumin from the damage induced by hydroxyl radicals effectively and even can form a kind of protein complex with human serum albumin showing more thermal stability. PMID- 18510338 TI - Preclinical studies to understand nanoparticle interaction with the immune system and its potential effects on nanoparticle biodistribution. AB - Nanoparticles have unique physicochemical properties which make them promising platforms for drug delivery. However, immune cells in the bloodstream (such as monocytes, platelets, leukocytes, and dendritic cells) and in tissues (such as resident phagocytes) have a propensity to engulf and eliminate certain nanoparticles. A nanoparticle's interaction with plasma proteins (opsonins) and blood components (via hemolysis, thrombogenicity and complement activation) may influence uptake and clearance and hence potentially affect distribution and delivery to the intended target sites. Nanoparticle uptake by the immune cells is influenced by many factors. Different nanoparticles have been shown to act on different pathways, while various characteristics/properties also affect which pathway is employed for particle internalization. Nanoparticle protein binding occurs almost instantaneously once the particle enters biological medium, and the physical properties of such a particle-protein complex are often different than those of the formulated particle. These new properties can contribute to different biological responses and change nanoparticle biodistribution. Therefore, in the situation when specific delivery to immune cells is not desired, the ideal nanoparticle platform is the one whose integrity is not disturbed in the complex biological environment, which provides extended circulation in the blood to maximize delivery to the target site, is not toxic to blood cellular components, and is "invisible" to the immune cells which can remove it from circulation. This review discusses the most recent data on nanoparticle interactions with blood components and how particle size and surface charge define their hematocompatibility. This includes properties which determine particle interaction with plasma proteins and uptake by macrophages. We will also provide an overview of in vitro methods useful in identifying interactions with components of the immune system and the potential effects of such interaction on particle distribution to tissues. PMID- 18510339 TI - Biochemical analysis of MST1 kinase: elucidation of a C-terminal regulatory region. AB - The MST1 kinase phosphorylates FoxO transcription factors in the cytosol and histone H2B in the nucleus to promote cellular apoptosis. In addition to a N terminal kinase domain, MST1 contains C-terminal regulatory and dimerization regions that are cleaved upon nuclear transport. In this report, we investigate the role of the MST1 regulatory region and dimerization domain in MST1 activity toward FoxO and histone H2B substrates. We find that the MST1 regulatory region enhances FoxO phosphorylation while inhibiting histone H2B phosphorylation, consistent with the cellular properties of MST1. We also identify autophosphorylation sites within the MST1 regulatory region and show that both regulatory region phosphorylation and MST1 dimerization contribute to FoxO phosphorylation. Together, our studies provide new insights into how MST1 substrate selectivity is modulated with implications for understanding apoptotic signaling through MST1 kinase. PMID- 18510340 TI - Bright and monodispersed phosphorescent particles and their applications for biological assays. AB - Halogen-containing polymers and copolymers have been discovered to provide excellent encapsulation matrixes for making bright and monodispersed phosphorescent nanoparticles. The phosphorescent nanoparticles exhibit strong phosphorescence with long lifetime and large Stoke shift under ambient conditions. The cross-linked phosphorescent particles using halogen-containing copolymers have been found to be very stable and easily resuspendable in aqueous media. The surface functional groups have been demonstrated to allow covalent tagging of biological recognition molecules such as antibodies to make particle antibody conjugates. The conjugates can be used to provide very sensitive detection of analytes through time-resolved phosphorescence measurements. PMID- 18510341 TI - Microfluidic-based cell sorting of Francisella tularensis infected macrophages using optical forces. AB - We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (microFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures (<4 ms) of focused 1064-nm laser light (9.6 W at the sample). We found no significant difference in viability, cell proliferation, activation state, and functionality between infrared-exposed and unexposed cells. Activation state was measured by the phosphorylation of ERK and nuclear translocation of NF-kappaB, while functionality was assessed in a similar manner, but after a lipopolysaccharide challenge. To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10,738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery. PMID- 18510342 TI - Multivariate analysis applied to the study of spatial distributions found in drug eluting stent coatings by confocal Raman microscopy. AB - Multivariate data analysis was applied to confocal Raman measurements on stents coated with the polymers and drug used in the CYPHER Sirolimus-eluting Coronary Stents. Partial least-squares (PLS) regression was used to establish three independent calibration curves for the coating constituents: sirolimus, poly(n butyl methacrylate) [PBMA], and poly(ethylene-co-vinyl acetate) [PEVA]. The PLS calibrations were based on average spectra generated from each spatial location profiled. The PLS models were tested on six unknown stent samples to assess accuracy and precision. The wt % difference between PLS predictions and laboratory assay values for sirolimus was less than 1 wt % for the composite of the six unknowns, while the polymer models were estimated to be less than 0.5 wt % difference for the combined samples. The linearity and specificity of the three PLS models were also demonstrated with the three PLS models. In contrast to earlier univariate models, the PLS models achieved mass balance with better accuracy. This analysis was extended to evaluate the spatial distribution of the three constituents. Quantitative bitmap images of drug-eluting stent coatings are presented for the first time to assess the local distribution of components. PMID- 18510343 TI - Dynamic component chemiluminescent sensor for assessing circulating polymorphonuclear leukocyte activity of peritoneal dialysis patients. AB - Recurrent bacterial peritonitis is a major complication in peritoneal dialysis (PD) patients, which is associated with polymorphonuclear leukocyte (PMN) functional changes and can be assessed by a chemiluminescent (CL) reaction. We applied a new approach of a dynamic component chemiluminescence sensor for the assessment of functional states of PMNs in a luminol-amplified whole-blood system. This method is based on the evaluation of CL kinetic patterns of stimulated PMNs, while the parallel measurements of intracellular and extracellular production of reactive oxygen species (ROS) from the same sample can be conducted. Blood was drawn from diabetic and nondiabetic patients during follow-up, and during peritonitis. Healthy medical personnel served as the control group. Chemiluminescence curves were recorded and presented as a sum of three biological components. CL kinetic parameters were calculated, and functional states of PMNs were assessed. Data mining algorithms were used to build decision tree models that can distinguish between different clinical groups. The induced classification models were used afterward for differentiating and classifying new blind cases and demonstrated good correlation with medical diagnosis (84.6% predictive accuracy). In conclusion, this novel method shows a high predictive diagnostic value and may assist in detection of PD-associated clinical states. PMID- 18510344 TI - Plasma-deposited fluorocarbon films: insulation material for microelectrodes and combined atomic force microscopy-scanning electrochemical microscopy probes. AB - Pinhole-free insulation of micro- and nanoelectrodes is the key to successful microelectrochemical experiments performed in vivo or in combination with scanning probe experiments. A novel insulation technique based on fluorocarbon insulation layers deposited from pentafluoroethane (PFE, CF3CHF2) plasmas is presented as a promising electrical insulation approach for microelectrodes and combined atomic force microscopy-scanning electrochemical microscopy (AFM-SECM) probes. The deposition allows reproducible and uniform coating, which is essential for many analytical applications of micro- and nanoelectrodes such as, e.g., in vivo experiments and SECM experiments. Disk-shaped microelectrodes and frame-shaped AFM tip-integrated electrodes have been fabricated by postinsulation focused ion beam (FIB) milling. The thin insulation layer for combined AFM-SECM probes renders this fabrication technique particularly useful for submicro insulation providing radius ratios of the outer insulation versus the disk electrode (RG values) suitable for SECM experiments. Characterization of PFE insulated AFM-SECM probes will be presented along with combined AFM-SECM approach curves and imaging. PMID- 18510345 TI - Self-modeling curve resolution recovery of temporal metabolite signal modulation in NMR spectroscopic data sets: application to a life-long caloric restriction study in dogs. AB - A novel model-free statistical approach (self modeling curve resolution, SMCR) has been applied to recover biochemical information from complex overlapping signals in (1)H NMR spectra of blood serum in a long-term study of caloric restriction (CR) in the dog (n = 24 control fed (CF) and n = 24 CR animals). A new statistical spectroscopic construct, the spectrotype, is proposed which is a spectroscopic subset description or component of a metabolic phenotype. Characterization of the (1)H NMR profiles according to their evolutionary contribution of each spectrotype gives clues to the kinetics of the macro biochemical response profiles and the identity of the underlying biochemical constituents, governing the evolutionary global response to an intervention. This information can be used to monitor and predict the end point of the biological process and to identify the mechanisms responsible for those changes. Here a SMCR strategy together with a pattern recognition method, principal component analysis (PCA) was used to resolve sets of spectrotypes, without a priori information. From the (1)H NMR evolutionary response profiles, two spectrotypes were identified and resolved; spectrotype 1 dominated by lipids featuring contributions from phosphatidylcholine, lipoprotein lipid fatty acyl groups from triglycerides, phospholipids, and cholesteryl esters plus total cholesterol (i.e., both esterified and unesterified); spectrotype 2 comprising glucose signals and a poorly resolved envelope of albumin and N-acetylated glycoprotein resonances. The relative contributions of these spectrotypes in each sample were calculated. For both caloric restricted (CR) and control fed (CF) dogs between ages 1 and 9 years, the contribution of spectrotype 2 > spectrotype 1, whereas for dogs aged between 9 and 12 years spectrotype 1 > spectrotype 2. Therefore, SMCR analysis pinpointed ages where nutrition and aging metabolic changes became significant within serum samples as well as providing the individual longitudinal contribution profiles associated with each spectrotype, which could potentially be used as part of a strategy to monitor and predict longevity and morbidity in populations. Hence SMCR is a useful addition to the chemometric "toolbox" for metabolic analysis and should have diverse applications within other biomedical conditions characterized by subtle time-dependent changes. PMID- 18510346 TI - Electrophoretic analysis of the mitochondrial outer membrane rupture induced by permeability transition. AB - A pathological increase of the permeability of the mitochondrial membranes may culminate in the irreversible rupture of the mitochondrial outer membrane. Such a permeability transition is lethal because it results in the release of death inducing molecules from mitochondria and/or metabolic failure. Current methods to assess this outer membrane damage are mostly indirect or scarcely representative of the overall mitochondrial population. Here we present an analytical and preparative approach using free flow electrophoresis to directly distinguish rat liver mitochondria that have undergone the permeability transition from unaffected organelles or from organelles that are damaged to a minor degree. Mitochondrial populations, which considerably differ in outer membrane integrity or cytochrome c content, were separated by this means. We further show that the relative abundance of each population depends on the dose of the permeability transition inducer and the duration of the treatment time. Finally, we have employed this approach to investigate the impairment of mitochondria that were isolated from livers subjected to ischemia/reperfusion damage. PMID- 18510347 TI - Detection of native protein ions in aqueous solution under ambient conditions by electrospray laser desorption/ionization mass spectrometry. AB - Liquid electrospray laser desorption/ionization (ELDI) mass spectrometry allows desorption and ionization of proteins directly from aqueous solutions and biological fluids under ambient conditions. Native protein ions such as those of myoglobin, cytochrome c, and hemoglobin were obtained. A droplet (ca. 5 microL) containing the protein molecules and micrometer-sized particles (e.g., carbon graphite powder) is irradiated with a pulsed UV laser. The laser energy adsorbed by the inert particles is transferred to the surrounding solvent and protein molecules, leading to their desorption; the desorbed gaseous molecules are then postionized within an electrospray (ESI) plume to generate the ESI-like protein ions. With the use of this technique, we detected only the protonated protein ions in various biological fluids (including human tears, cow milk, serum, and bacterial extracts) without interference from their corresponding sodiated or potassiated adduct ions. In addition, we rapidly quantified the levels of glycosylated hemoglobin present in drops of whole blood obtained from diabetic patients without the need of sample pretreatment. PMID- 18510348 TI - Wavelet-based method for noise characterization and rejection in high-performance liquid chromatography coupled to mass spectrometry. AB - We present a new method for rejecting noise from HPLC-MS data sets. The algorithm reveals peptides at low concentrations by minimizing both the chemical and the random noise. The goal is reached through a systematic approach to characterize and remove the background. The data are represented as two-dimensional maps, in order to optimally exploit the complementary dimensions of separation of the peptides offered by the LC-MS technique. The virtual chromatograms, reconstructed from the spectrographic data, have proved to be more suitable to characterize the noise than the raw mass spectra. By means of wavelet analysis, it was possible to access both the chemical and the random noise, at different scales of the decomposition. The novel approach has proved to efficiently distinguish signal from noise and to selectively reject the background while preserving low abundance peptides. PMID- 18510349 TI - Identification of disulfide-containing chemical cross-links in proteins using MALDI-TOF/TOF-mass spectrometry. AB - Cross-linking can be used to identify spatial relationships between amino acids in proteins or protein complexes. A rapid and sensitive method for identifying the site of protein cross-linking using dithiobis(sulfosuccinimidyl propionate) (DTSSP) is presented and illustrated with experiments using murine cortactin, actin and acyl-CoA thioesterase. A characteristic 66 Da doublet, which arises from the asymmetric fragmentation of the disulfide of DTSSP-modified peptides, is observed in the mass spectra obtained under MALDI-TOF/TOF-MS conditions and allows rapid assignment of cross-links in modified proteins. This doublet is observed not only for linear cross-linked peptides but also in the mass spectra of cyclic cross-linked peptides when simultaneous fragmentation of the disulfide and the peptide backbone occurs. We suggest a likely mechanism for this fragmentation. We use guanidinylation of the cross-linked peptides with O-methyl isourea to extend the coverage of cross-linked peptides observed in this MALDI-MS technique. The methodology we report is robust and amenable to automation, and permits the analysis of native cystines along with those introduced by disulfide containing cross-linkers. PMID- 18510350 TI - Synthesis and biological characterization of novel 2-quinolinecarboxamide ligands of the peripheral benzodiazepine receptors bearing technetium-99m or rhenium. AB - Potential receptor imaging agents based on Tc-99m for the in vivo visualization of the peripheral benzodiazepine receptor (PBR) have been designed on the basis of the information provided by the previously published structure-affinity relationship studies, which suggested the existence of tolerance to voluminous substituents in the receptor area interacting with 3-position of the quinoline nucleus of 2-quinolinecarboxamides 5. In the first step of the investigation, the stereoelectronic features of the above-indicated receptor area were also probed by means of 4-phenyl-3-[(1-piperazinyl)methyl]-2-quinolinecarboxamide derivatives bearing different substituents on the terminal piperazine nitrogen atom (compounds 6a-f). The structure-affinity relationship data confirmed the existence of a tolerance to bulky lipophilic substituents and stimulated the design of bifunctional ligands based on the 4-phenyl-3-[(1-piperazinyl)methyl]-2 quinolinecarboxamide moiety (compounds 6h,j,k,m). The submicromolar PBR affinity of rhenium complexes 6j,m suggests that the presence of their metal-ligand moieties with encaged rhenium is fairly compatible with the interaction with the PBR binding site. Thus, in order to obtain information on the in vivo behavior of these bifunctional ligands, (99m)Tc-labeled compounds 6h,k were synthesized and evaluated in preliminary biodistribution and single photon emission tomography (SPET) studies. The results suggest that both tracers do not present a clear preferential distribution in tissues rich in PBR, probably because of their molecular dimensions, which may hamper both the intracellular diffusion toward PBR and the interaction with the binding site. PMID- 18510351 TI - Development of novel 68Ga- and 18F-labeled GnRH-I analogues with high GnRHR targeting efficiency. AB - A large majority of tumors of the reproductive system express the gonadotropin releasing hormone receptor (GnRHR). Blockade and activation of this receptor with various antagonistic and agonistic analogues of native GnRH-I (pGlu(1)-His(2) Trp(3)-Ser(4)-Tyr(5)-Gly (6)-Leu(7)-Arg(8)-Pro(9)-Gly(10)-NH2), respectively, has shown efficient suppression of tumor growth. In this study, the GnRH-receptor system has been evaluated with respect to its suitability as a target for in vivo peptide receptor targeting using radiolabeled GnRH-analogues, and in parallel, new (18)F- and (68Ga)-labeled GnRH analogues have been developed. In vitro radioligand binding assays performed with various GnRHR-expressing human cell lines using [(125)I]Triptorelin (D-Trp(6)-GnRH-I) as the standard radioligand revealed a very low level of GnRH receptor expression on the cell surface. Generally, total cellular activity was very low (approximately 3% of the applied activity), and only a small fraction (max. 40%) of cell-associated activity could be attributed to receptor-specific radioligand binding/internalization. However, substitution of fetal calf serum by NU serum in the culture medium led to increased and stable GnRHR-expression, especially in the ovarian cancer cell line EFO-27, thus allowing for a stable experimental setup for the evaluation of the new radiolabeled GnRH-I analogues. The new radiolabeled GnRH-I analogues developed in this study were all based on the D-Lys(6)-GnRH-I-scaffold. For (68)Ga-labeling, the latter was coupled with DOTA at D-Lys(6). To allow (18)F labeling via chemoselective oxime formation, D-Lys(6)-GnRH-I was also conjugated with Ahx (aminohexanoic acid) or beta-Ala, which in turn was coupled with Boc aminooxyacetic acid. (18)F-labeling via oxime formation with 4 [(18)F]fluorobenzaldehyde was performed using the Boc-protected precursors. Receptor affinities of [(68)Ga]DOTA-GnRH-I, D-Lys(6)-Ahx([(18)F]FBOA)-GnRH-I, and D-Lys(6)-betaAla([(18)F]FBOA)-GnRH-I (FBOA = fluorobenzyloxime acetyl) were determined using GnRHR-membrane preparations, and internalization efficiency of the new radioligands was determined in EFO-27 cells. Both quantities were highest for D-Lys(6)-Ahx([(18)F]FBOA)-GnRH-I (IC 50 = 0.50 +/- 0.08 nM vs 0.13 +/- 0.08 nM for Triptorelin; internalization: 86 +/- 16% of the internal reference [(125)I]Triptorelin), already substantially reduced in the case of the betaAla([(18)F]FBOA)-derivative (IC 50 = 0.86 +/- 0.13 nM; internalization: 42 +/ 3% of [(125)I]Triptorelin), while the [(68)Ga]DOTA-analogue showed almost complete loss of binding affinity and ligand internalization (IC50 = 13.3 +/- 1.0 nM; internalization: 2.6 +/- 1.0% of [(125)I]Triptorelin). Generally, the lipophilic residue [(18)F]FBOA is much better tolerated as a modification of the D-Lys(6)-side chain, with receptor affinity of the respective analogues strongly depending upon spacer length between the D-Lys(6)-side chain and the [(18)F]FBOA moiety. In summary, D-Lys(6)(Ahx-[(18)F]FBOA)-GnRH-I shows the highest potential for efficient GnRHR-targeting in vivo of the compounds investigated. Unfortunately, however, the very low cell surface expression of GnRH-receptors and thus very low radioligand uptake by GnRHR-positive tumor cells found in vitro was also confirmed by a preliminary biodistribution study in OVCAR-3 xenografted nude mice using the standard GnRHR radioligand [(125)I]Triptorelin. Tumor uptake was lower than blood activity concentration at 1 h p.i. (0.49 +/- 0.05 vs 0.96 +/ 0.13 for tumor and blood, respectively). These data seriously challenge the suitability of the GnRHR-system as a suitable target for in vivo peptide receptor imaging using radiolabeled GnRH-I derivatives, despite the availability of high affinity radiolabeled receptor-ligands such as D-Lys(6)(Ahx-[(18)F]FBOA)-GnRH-I. PMID- 18510352 TI - How changes in the sequence of the peptide CLPFFD-NH2 can modify the conjugation and stability of gold nanoparticles and their affinity for beta-amyloid fibrils. AB - In a previous work, we studied the interaction of beta-amyloid fibrils (Abeta) with gold nanoparticles (AuNP) conjugated with the peptide CLPFFD-NH2. Here, we studied the effect of changing the residue sequence of the peptide CLPFFD-NH2 on the efficiency of conjugation to AuNP, the stability of the conjugates, and the affinity of the conjugates to the Abeta fibrils. We conjugated the AuNP with CLPFFD-NH 2 isomeric peptides (CDLPFF-NH2 and CLPDFF-NH2) and characterized the resulting conjugates with different techniques including UV-Vis, TEM, EELS, XPS, analysis of amino acids, agarose gel electrophoresis, and CD. In addition, we determined the proportion of AuNP bonded to the Abeta fibrils by ICP-MS. AuNP CLPFFD-NH2 was the most stable of the conjugates and presented more affinity for Abeta fibrils with respect to the other conjugates and bare AuNP. These findings help to better understand the way peptide sequences affect conjugation and stability of AuNP and their interaction with Abeta fibrils. The peptide sequence, the steric effects, and the charge and disposition of hydrophilic and hydrophobic residues are crucial parameters when considering the design of AuNP peptide conjugates for biomedical applications. PMID- 18510353 TI - Scaled-down purification protocol to access proteomic analysis of 20S proteasome from human tissue samples: comparison of normal and tumor colorectal cells. AB - The proteasome is a proteolytic complex that constitutes the main pathway for degradation of intracellular proteins in eukaryotic cells. It regulates many physiological processes and its dysfunction can lead to several pathologies like cancer. To study the 20S proteasome structure/activity relationship in cells that derive from human biopsy samples, we optimized an immuno-purification protocol for the analysis of samples containing a small number of cells using magnetic beads. This scaled-down protocol was used to purify the cytoplasmic 20S proteasome of adjacent normal and tumor colorectal cells arising from tissue samples of several patients. Proteomic analyses based on two-dimensional gel electrophoresis (2DE) and mass spectrometry showed that the subunit composition of 20S proteasomes from these normal and tumor cells were not significantly different. The proteasome activity was also assessed in the cytoplasmic extracts and was similar or higher in tumor colorectal than in the corresponding normal cells. The scaled-down 20S proteasome purification protocol developed here can be applied to any human clinical tissue samples and is compatible with further proteomic analyses. PMID- 18510354 TI - Vegetable proteomics: the detection of Ole e 1 isoallergens by peptide matching of MALDI MS/MS spectra of underivatized and dansylated glycopeptides. AB - Ole e 1 (NCBI entry gi|14424429) is the major allergen of Oleaceae family. Multiple isoforms and variants are present in varying degrees of distribution. In this report, we present a new approach to the resolution of multiple forms of Ole e 1 from whole antigen extracts, based on a preliminary chemical fractionation procedure followed by MALDI MS and MS/MS measurements. The characterization of Ole e 1 isoallergens was accomplished through the identification of the amino acid sequence including the glycosylation site and the structure of the glycan moieties. The structure feature of the identified Ole e 1.0102 (gi|2465127), main olive allergen [Olea europaea] (gi|13195753), Major pollen allergen Ole e 1 (gi|33329740) and Ole e 1c (gi|1362131) is represented by the point mutation K(106) --> I and by the presence of a glycan moiety. Two other variants Major pollen allergen (Allergen Ole e1) (Ole e I) (gi|14424429) and Ole e 1.0103 protein [Olea europea] (gi|2465129) were identified as nonglycosylated species. These results, partially in disagreement with Swiss-Prot annotation, were validated by matching the MALDI MS/MS spectra of the natural tryptic mixture with those obtained after deglycosylation. PMID- 18510355 TI - Phosphoproteomic analysis of human brain by calcium phosphate precipitation and mass spectrometry. AB - Alzheimer's disease (AD), the most common form of dementia, is manifested in the brain by the aggregation of amyloid plaques and neurofibrillary tangles. The tangles are primarily composed of microtubule-associated protein tau that is aberrantly hyperphosphorylated, suggesting that deregulated phosphorylation may contribute to AD pathogenesis. However, systematic analysis of the phosphoproteome in AD brain tissues has not been reported. We used calcium phosphate precipitation to analyze an AD postmortem brain, followed by liquid chromatography-tandem mass spectrometry. The protein sample was first resolved by one-dimensional polyacrylamide gel electrophoresis and subjected to gel excision and in-gel digestion. Phosphopeptides in the resulting peptide mixtures were enriched in a single step of calcium phosphate precipitation, and then analyzed by the LC-MS/MS approach. After database search, stringent filtering, and manual validation of neutral loss in the MS/MS spectra, a total of 466 phosphorylation sites on 185 proteins including tau were identified. A majority of sites were not described previously. This study demonstrates the feasibility of combining calcium phosphate precipitation with mass spectrometry for phosphoproteome analysis of postmortem human brain tissue. PMID- 18510356 TI - Phosphoprotein profiling by PA-GeLC-MS/MS. AB - A significant consequence of protein phosphorylation is to alter protein-protein interactions, leading to dynamic regulation of the components of protein complexes that direct many core biological processes. Recent proteomic studies have populated databases with extensive compilations of cellular phosphoproteins and phosphorylation sites and a similarly deep coverage of the subunit compositions and interactions in multiprotein complexes. However, considerably less data are available on the dynamics of phosphorylation, composition of multiprotein complexes or that define their interdependence. We describe a method to identify candidate phosphoprotein complexes by combining phosphoprotein affinity chromatography, separation by size, denaturing gel electrophoresis, protein identification by tandem mass spectrometry, and informatics analysis. Toward developing phosphoproteome profiling, we have isolated native phosphoproteins using a phosphoprotein affinity matrix, Pro-Q Diamond resin (Molecular Probes-Invitrogen). This resin quantitatively retains phosphoproteins and associated proteins from cell extracts. Pro-Q Diamond purification of a yeast whole cell extract followed by 1-D PAGE separation, proteolysis and ESI LC-MS/MS, a method we term PA-GeLC-MS/MS, yielded 108 proteins, a majority of which were known phosphoproteins. To identify proteins that were purified as parts of phosphoprotein complexes, the Pro-Q eluate was separated into two fractions by size, <100 kDa and >100 kDa, before analysis by PAGE and ESI LC-MS/MS and the component proteins queried against databases to identify protein-protein interactions. The <100 kDa fraction was enriched in phosphoproteins indicating the presence of monomeric phosphoproteins. The >100 kDa fraction contained 171 proteins of 20-80 kDa, nearly all of which participate in known protein-protein interactions. Of these 171, few are known phosphoproteins, consistent with their purification by participation in protein complexes. By comparing the results of our phosphoprotein profiling with the informational databases on phosphoproteomics, protein-protein interactions and protein complexes, we have developed an approach to examining the correlation between protein interactions and protein phosphorylation. PMID- 18510357 TI - 18O labeling over a coffee break: a rapid strategy for quantitative proteomics. AB - Proteomics-based quantification methods for differential protein expression measurements are among the most important and challenging techniques in the field of mass spectrometry. Though numerous quantification methods have been established, no method meets all the demands for measuring accurate protein expression levels. Of the various relative quantification methods by isotopic labeling, (18)O labeling method has been shown to be simple, specific, cost effective and applicable to a wide range of analyses. However, some researchers refrain from using the method due to long incubation periods required during the labeling process. To address this problem, we demonstrate a method by which the labeling procedure can be completed in 15 min. We digested and labeled samples using immobilized trypsin on micro-spin columns to speed up the enzyme-mediated oxygen substitution, thereby completing the labeling process within 15 min with high labeling efficiency. We demonstrate the efficiency and accuracy of the method using a four protein mixture and whole cell lysate from rat vascular endothelial cells. PMID- 18510358 TI - Comparison of SYPRO Ruby and Deep Purple using commonly available UV transilluminator: wide-scale application in proteomic research. AB - Fluorescent stains are becoming increasingly useful in proteomics research involving protein expression as well as post-translational modification studies and are particularly useful for samples which are expensive and scarce. The fluorescent dyes Deep Purple and SYPRO Ruby are widely used in protein expression studies. Using UV transillumination and Charged Coupled Device (CCD) based imaging system, their relative sensitivity to detect proteins separated by two dimensional polyacrylamide gel electrophoresis and downstream protein identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was compared. Using mouse liver homogenate, we detected a greater number of spots using SYPRO Ruby over Deep Purple stain. However, the number of matched peptides and the percentage of amino acid residues identified for 21 different proteins were comparable suggesting their equivalency for LC-MS/MS identification. In spite of comparable MS compatibility, we recommend the use of SYPRO Ruby for expression proteomics due to its higher sensitivity in detecting protein spots. PMID- 18510359 TI - Facile, efficient approach to accomplish tunable chemistries and variable biodistributions for shell cross-linked nanoparticles. AB - The in vivo behavior of shell cross-linked knedel-like (SCK) nanoparticles is shown to be tunable via a straightforward and versatile process that advances SCKs as attractive nanoscale carriers in the field of nanomedicine. Tuning of the pharmacokinetics was accomplished by grafting varied numbers of methoxy terminated poly(ethylene glycol) (mPEG) chains to the amphiphilic block copolymer precursors, together with chelators for the radioactive tracer and therapeutic agent (64)Cu, followed by self-assembly into block copolymer micelles and chemical cross-linking throughout the shell regions. (64)Cu-radiolabeling was then performed to evaluate the SCKs in vivo by means of biodistribution experiments and positron emission tomography (PET). It was found that the blood retention of PEGylated SCKs could be tuned, depending on the mPEG grafting density and the nanoparticle surface properties. A semiquantitative model of the density of mPEG surface coverage as a function of in vivo behavior was applied to enhance the understanding of this system. PMID- 18510360 TI - Highly filled bionanocomposites from functionalized polysaccharide nanocrystals. AB - Cellulose and starch nanocrystals obtained from the acid hydrolysis of ramie fibers and waxy maize starch granules, respectively, were subjected to isocyanate mediated reaction to graft polycaprolactone (PCL) chains with various molecular weights on their surface. Grafted nanoparticles were characterized by X-ray diffraction analysis and contact angle measurements. We observed that the nanoparticles kept their initial morphological integrity and native crystallinity. Nanocomposite films were processed from both unmodified and PCL grafted nanoparticles and PCL as matrix using a casting/evaporation technique. We showed that mechanical properties of resulting films were notably different. Compared to unmodified nanoparticles, the grafting of PCL chains on the surface results in lower modulus values but significantly higher strain at break. This unusual behavior clearly reflects the originality of the reinforcing phenomenon of polysaccharide nanocrystals resulting from the formation of a percolating network thanks to chain entanglements and cocrystallization. PMID- 18510361 TI - Liquid crystalline phase behavior of high molecular weight DNA: a comparative study of the influence of metal ions of different size, charge and binding mode. AB - The ability of Li(+), Na(+), K(+), Rb(+), Cs(+), Mg(2+), Ca(2+), Sr(2+), Ba(2+), Cu(2+), Cd(2+), Al(3+), V(4+), Hg(2+), Pd(2+), Au(3+), and Pt(4+) to provoke liquid crystalline (LC) phases in high molecular weight DNA was investigated. The alkali and alkaline earth metal ions provoked typical cholesteric/columnar structures, whereas transition metal ions precipitated DNA into solid/translucent gel-like aggregates. Heavy metal ions reduced viscosity of DNA solution, disrupting rigid, rod-like DNA structure necessary for LC textures. Three-layer quantum mechanical-molecular mechanical (QM/MM) studies of Li(+), Na(+), K(+), Mg(2+), and Ca(2+) binding DNA fragment suggested several possible binding modes of these ions to the phosphate groups. The dianion mode of metal binding, involving the phosphate groups of both strands of DNA, allowed for higher DNA binding affinity of the alkaline earth metal ions. These results have implications in understanding the biological role of metal ions and developing DNA-based sensors and nanoelectronic devices. PMID- 18510362 TI - Bioactive isocoumarins isolated from the endophytic fungus Microdochium bolleyi. AB - Three new isocoumarin derivatives ( 2- 4) were isolated together with monocerin ( 1) from Microdochium bolleyi, an endophytic fungus from Fagonia cretica, a herbaceous plant of the semiarid coastal regions of Gomera. Compounds 2 and 3 are both 12-oxo epimers of 1, and 4 is a ring-opened derivative of 1. The structures were elucidated by detailed spectroscopic analysis and comparison with reported data. The absolute configurations were determined by a modified Mosher's method. Compounds 1, 3, and 4 showed good antifungal, antibacterial, and antialgal activities against Microbotryum violaceum, Escherichia coli, Bacillus megaterium, and Chlorella fusca. Compound 2 was moderately antifungal and antialgal. PMID- 18510363 TI - Solid-phase synthesis of asymmetrically substituted "AB3-type" phthalocyanines. AB - Synthesis of phthalocyanines with asymmetrical substitution on the periphery is often difficult due the problems in purification of the phthalocyanine mixtures obtained. Using a poly(ethylene glycol) (PEG)-based support with a Wang-type linker, we have developed the synthesis of monohydroxylated, oligoethylene glycol substituted phthalocyanines utilizing an amidine-base-promoted phthalonitrile tetramerization reaction. The use of a hydrophilic support allows symmetrical phthalocyanine product formed in solution to be readily and completely removed by washing while leaving the "AB3" product on the support. Acid cleavage with 10% trifluoroacetic acid provides the pure unsymmetrically substituted Pc. This method was applied to several metallo Pcs. Additionally, methods to avoid premature reactions on-resin that give A2B2 products are provided. PMID- 18510364 TI - Aryl-aryl bond formation by flash vacuum pyrolysis of benzannulated thiopyrans. AB - In contrast to fully unsaturated 7-membered ring sulfur heterocycles (thiepines), some of which extrude sulfur and give the ring-contracted hydrocarbon even at room temperature in solution, benzannulated thiopyrans (6-membered sulfur heterocycles) require flash vacuum pyrolysis (FVP) conditions in the gas phase at temperatures in the range of 1000-1200 degrees C to promote the corresponding reaction. Thus, FVP of benzo[kl]thioxanthene (1) gives fluoranthene, and naphtho[2,1,8,7-klmn]thioxanthene (6) gives benzo[ghi]fluoranthene (7). FVP of thioxanthone (9) gives fluorenone (10), together with lesser amounts of dibenzo[b,d]thiophene (11), from competing decarbonylation. PMID- 18510365 TI - Enantiomeric resolution of N,N'-dimethyldithiodianthranilide through diastereomeric silver(I) complex. Circular dichroism spectra, racemization barrier, and molecular self-assembly. AB - Planar chiral N,N'-dimethyldithiodianthranilide (2b) was resolved to enantiomers through a diasteromeric complex with easily accessible silver(I) (1S) camphorosulfonate (3). The (-)-2b enantiomer was assigned the R absolute configuration from the X-ray crystal structure of the silver complex. The compound is configurationally stable and its racemization occurs through boat-to boat ring inversion (DeltaG(double dagger) = 36.5 +/- 0.2 kcal mol(-1) at 438 K). The analysis of the CD spectrum of the title compound showed that the n-pi* Cotton effect sign is determined by the helicity of the skewed thiobenzamide chromophore. The molecules of 2b are unable to achieve efficient crystal packing by themselves and easily form inclusion complexes with toluene or pentafluorophenol. PMID- 18510366 TI - Structure-based drug design: exploring the proper filling of apolar pockets at enzyme active sites. AB - The proper filling of apolar pockets at enzyme active sites is central for increasing binding activity and selectivity of hits and leads in medicinal chemistry. In our structure-based design approach toward the generation of potent enzyme inhibitors, we encountered a variety of challenges in gaining suitable binding affinity from the occupation of such pockets. We summarize them here for the first time. A fluorine scan of tricyclic thrombin inhibitors led to the discovery of favorable orthogonal dipolar C-F...CO interactions. Efficient cation pi interactions were established in the S4 pocket of factor Xa, another serine protease from the blood coagulation cascade. Changing from mono- to bisubstrate inhibitors of catechol O-methyltransferase, a target in the L-Dopa-based treatment of Parkinson's disease, enabled the full exploitation of a previously unexplored hydrophobic pocket. Conformational preorganization of a pocket at an enzyme active site is crucial for harvesting binding affinity. This is demonstrated for two enzymes from the nonmevalonate pathway of isoprenoid biosynthesis, IspE and IspF, which are pursued as antimalarial targets. Disrupting crystallographically defined water networks on the way into a pocket might cost all of the binding free enthalpy gained from its occupation, as revealed in studies with tRNA-guanine transglycosylase, a target against shigellosis. Investigations of the active site of plasmepsin II, another antimalarial target, showed that principles for proper apolar cavity filling, originally developed for synthetic host-guest systems, are also applicable to enzyme environments. PMID- 18510367 TI - Maximizing efficiency in the production of compound libraries. AB - Efficiency is one of the most important criteria in departments responsible for the production of compounds in a library format. Consequently, this was a key factor in the initial design of our automated medicinal chemistry department, established some years ago. Nonetheless, we were able to improve and optimize our workflows and processes constantly. Here, we outline our current setup, from design to submission of libraries, and discuss which procedures and techniques appear to be useful for us and which ones turned out to be less effective. The aim of the manuscript is not to present individualized and tailor-made solutions in our laboratory but rather to describe approaches (often executed with commercial equipment) which might be of relevance for a broader readership working in this field. PMID- 18510368 TI - Exciton and biexciton luminescence from single GaN/AlN quantum dots in nanowires. AB - We present a microphotoluminescence study of single GaN/AlN quantum dots embedded in single nanowires. At low excitation power, single exciton lines with full width at half-maximum as narrow as 1 meV are observed. The study of the excitation power dependence of the emission allows us to identify the biexciton transitions with binding energies ranging from 20 to 40 meV. PMID- 18510369 TI - Nature of the second optical transition in PbSe nanocrystals. AB - The second peak in the optical absorption spectrum of PbSe nanocrystals is arguably the most discussed optical transition in semiconductor nanocrystals. Ten years of scientific debate have produced many theoretical and experimental claims for the assignment of this feature as the 1P e1P h as well as the 1S h,e1P e,h transitions. We studied the nature of this absorption feature by pump-probe spectroscopy, exactly controlling the occupation of the states involved, and present conclusive evidence that the optical transition involves neither 1S e nor 1S h states. This suggests that it is the 1P h1P e transition that gives rise to the second peak in the absorption spectrum of PbSe nanocrystals. PMID- 18510370 TI - Surface effect on the elastic behavior of static bending nanowires. AB - The surface effect from surface stress and surface elasticity on the elastic behavior of nanowires in static bending is incorporated into Euler-Bernoulli beam theory via the Young-Laplace equation. Explicit solutions are presented to study the dependence of the surface effect on the overall Young's modulus of nanowires for three different boundary conditions: cantilever, simply supported, and fixed fixed. The solutions indicate that the cantilever nanowires behave as softer materials when deflected while the other structures behave like stiffer materials as the nanowire cross-sectional size decreases for positive surface stresses. These solutions agree with size dependent nanowire overall Young's moduli observed from static bending tests by other researchers. This study also discusses possible reasons for variations of nanowire overall Young's moduli observed. PMID- 18510371 TI - Clavatadine A, a natural product with selective recognition and irreversible inhibition of factor XIa. AB - Bioassay-guided fractionation of a CH2Cl2/MeOH extract of the sponge Suberea clavata using the serine protease factor XIa to detect antithrombotic activity led to the isolation of the new marine natural products, clavatadines A and B. Clavatadines A and B inhibited factor XIa with IC50's of 1.3 and 27 microM, respectively. A crystal structure of protein-inhibitor (clavatadine A) complex was obtained and revealed interesting selective binding and irreversible inhibition of factor XIa. The cocrystal structure provides guidance for the design and synthesis of future factor XIa inhibitors as antithrombotic agents. PMID- 18510372 TI - Calculation of quantum-mechanical descriptors for QSPR at the DFT level: is it necessary? AB - Most of the recently published quantitative structure-property relationship (QSPR) models, which can be used to predict environmentally relevant physicochemical data for persistent organic pollutants (e.g., polychlorinated dibenzo- p-dioxins, dibenzofurans, and biphenyls), employ molecular descriptors obtained by means of relatively costly calculations at the density functional theory (DFT) level. However, new semiempirical methods, PM6 and RM1, have recently been developed by J. J. P. Stewart's group. In this study, we compared various QSPR models based on DFT (B3LYP functional) descriptors with the same models based on semiempirical (PM6 and RM1) descriptors. We recalibrated 10 previously published models (for different properties and groups of congeneric compounds) employing PM6 and RM1 descriptors instead of B3LYP ones. We demonstrated that by applying RM1 and PM6 descriptors, we could obtain QSPR models with quality similar to that of models based on B3LYP descriptors. This level of accuracy was out of reach for the models employing AM1- and PM3-based descriptors. PMID- 18510373 TI - Synthesis of thiophene-containing hybrid calixphyrins of the 5,10-porphodimethene type. AB - The synthesis, structure, and optical and electrochemical properties of thiophene containing hybrid calixphyrins are reported. The 5,10-porphodimethene type 14pi- and 16pi-S,N2,X-hybrid calixphyrins (X = NH, O, S) were prepared by acid-promoted dehydrative condensation between a thiatripyrrane and the corresponding 2,5 bis[hydroxy(phenyl)methyl]heteroles followed by DDQ oxidation. Both crystallographic and spectroscopic analyses of the newly prepared hybrid calixphyrins have revealed that the combination of heteroles explicitly influences the electronic structures of the pi-conjugated framework. The 14pi S,N2,X-hybrid calixphyrins have proven to be fluorescent in solution. PMID- 18510374 TI - Biopolymer microparticle and nanoparticle formation within a microfluidic device. AB - This paper reports a novel microfluidic method for the production of cross-linked alginate microparticles and nanoparticles. We describe a continuous process relying on both thermodynamic and hydrodynamic factors to form microdroplets. A rapid cross-linking reaction thereafter allows solidification of the polymer droplets either within the microfluidic device or "off-chip" to form alginate micro- and nanoparticles. Monodisperse droplets are generated by extruding an aqueous alginate solution using an axisymmetric flow-focusing design. As they flow downstream in the channel, due to water and the continuous phase being partially miscible, the water diffuses very slowly out of the polymeric droplets into the transport fluid, which causes the shrinkage of the drops and the condensation of the polymer phase. The resulting size of the solid particles depends on the polymer concentration and the ensuing balance between the kinetics of the cross-linking reaction and the volume loss due to solvent diffusion. This work details both a single-step microfluidic technique for the formation of alginate microparticles of sizes ranging from 1 to 50 microm via near-equilibrium solvent diffusion within a microfluidic device and thereafter a two-step method, which was shown to generate biopolymer nanoparticles of sizes ranging from 10 to 300 nm. These novel methodologies are extremely flexible and can be extended to the preparation of micro- and nanoparticles from a wide range of single or mixed synthetic and biologically derived polymers. PMID- 18510375 TI - Structure and activity assay of nanozymes prepared by the coimmobilization of practically useful enzymes and hydrophilic block copolymers on gold nanoparticles. AB - Enzyme/polymer/gold nanoparticle hybrids, called "nanozymes", were prepared and structurally analyzed by dynamic light scattering (DLS), ultraviolet-visible spectroscopy, and zeta-potential and transmission electron microscopy (TEM) measurements, which showed that the nanozyme particles were mainly composed of a single gold nanoparticle, on whose surface the enzyme and polymer were coimmobilized. This kind of structure resulted in the high dispersion stability of the nanozyme under various conditions, accompanied by improved thermal stability of the enzyme. PMID- 18510376 TI - Double cushions preserve transmembrane protein mobility in supported bilayer systems. AB - Supported lipid bilayers (SLBs) have been widely used as model systems to study cell membrane processes because they preserve the same 2D membrane fluidity found in living cells. One of the most significant limitations of this platform, however, is its inability to incorporate mobile transmembrane species. It is often postulated that transmembrane proteins reconstituted in SLBs lose their mobility because of direct interactions between the protein and the underlying substrate. Herein, we demonstrate a highly mobile fraction for a transmembrane protein, annexin V. Our strategy involves supporting the lipid bilayer on a double cushion, where we not only create a large space to accommodate the transmembrane portion of the macromolecule but also passivate the underlying substrate to reduce nonspecific protein-substrate interactions. The thickness of the confined water layer can be tuned by fusing vesicles containing polyethyleneglycol (PEG)-conjugated lipids of various molecular weights to a glass substrate that has first been passivated with a sacrificial layer of bovine serum albumin (BSA). The 2D fluidity of these systems was characterized by fluorescence recovery after photobleaching (FRAP) measurements. Uniform, mobile phospholipid bilayers with lipid diffusion coefficients of around 3 x 10(-8) cm2/s and percent mobile fractions of over 95% were obtained. Moreover, we obtained annexin V diffusion coefficients that were also around 3 x 10(-8) cm2/s with mobile fractions of up to 75%. This represents a significant improvement over bilayer platforms fabricated directly on glass or using single cushion strategies. PMID- 18510377 TI - Gastrointestinal stromal tumors: key to diagnosis and choice of therapy. AB - The common feature of gastrointestinal stromal tumors (GISTs) is the expression of KIT protein or acquisition of activating, constitutive mutations in the KIT or platelet-derived growth factor receptor alpha (PDGFRA) genes that are the early oncogenic events during GIST development. With these discoveries, GIST has emerged as a distinct sarcoma entity, enabling the introduction of targeted therapy using the inhibition of KIT/PDGFRA and their downstream signaling cascade. The introduction of a small-molecule tyrosine kinase inhibitor, imatinib mesylate, to clinical practice has revolutionized the treatment of patients with advanced GISTs and is currently approved as first-line treatment for patients with metastatic and/or inoperable GISTs. Mutation screening is currently a tool in GIST diagnosis, assessment of sensitivity to tyrosine kinase inhibitors, and prediction of achieving response to molecularly targeted therapy. This article discusses the histologic and molecular criteria for distinguishing GISTs from other types of sarcoma, and the molecular diagnostic tools that are currently available or in development to assist in therapy decisions. PMID- 18510378 TI - Cerebral small vessel disease: genetic risk assessment for prevention and treatment. AB - Cerebrovascular disease is a major burden to individuals and their communities worldwide. Stroke is one of the leading causes of death and disability, and the prevention and treatment of stroke can be improved with a better understanding of its causation. Cerebral small vessel disease (SVD) is a subset of cerebrovascular disease, and has an equally large impact on an individual's quality of life. Although many risk factors are involved, we propose that genetics has a significant role in the pathogenesis of SVD through a complex interplay of environmental and multigenetic factors. Advances in molecular technology have enabled the human genome to be investigated both at a population and, more recently, an individual level. A better understanding of the molecular basis of SVD will enable the development of therapies to help in its prevention and treatment. This review assesses the molecular genetics underlying cerebral SVD. PMID- 18510379 TI - Biomarkers for osteoporosis management: utility in diagnosis, fracture risk prediction and therapy monitoring. AB - Osteoporosis is a systemic disease characterized by low bone mass and microarchitectural deterioration of bone tissue, resulting in an increased risk of fracture. While the level of bone mass can be estimated by measuring bone mineral density (BMD) using dual X-ray absorptiometry (DXA), its measurement does not capture all the risk factors for fracture. Quantitative changes in skeletal turnover can be assessed easily and non-invasively by the measurement of serum and urinary biochemical markers; the most sensitive markers include serum osteocalcin, bone specific alkaline phosphatase, the N-terminal propeptide of type I collagen for bone formation, and the crosslinked C- (CTX) and N- (NTX) telopeptides of type I collagen for bone resorption. Advances in our knowledge of bone matrix biochemistry, most notably of post-translational modifications in type I collagen, are likely to lead to the development of new biochemical markers that reflect changes in the material property of bone, an important determinant of bone strength. Among those, the measurement of the urinary ratio of native (alpha) to isomerized (beta) CTX - an index of bone matrix maturation - has been shown to be predictive of fracture risk independently of BMD and bone turnover. In postmenopausal osteoporosis, levels of bone resorption markers above the upper limit of the premenopausal range are associated with an increased risk of hip, vertebral, and nonvertebral fracture, independent of BMD. Therefore, the combined use of BMD measurement and biochemical markers is helpful in risk assessment, especially in those women who are not identified as at risk by BMD measurement alone. Levels of bone markers decrease rapidly with antiresorptive therapies, and the levels reached after 3-6 months of therapy have been shown to be more strongly associated with fracture outcome than changes in BMD. Preliminary studies indicate that monitoring changes of bone formation markers could also be useful to monitor anabolic therapies, including intermittent parathyroid hormone administration and, possibly, to improve adherence to treatment. Thus, repeated measurements of bone markers during therapy may help improve the management of osteoporosis in patients. PMID- 18510381 TI - Molecular Medicine TriConference 2008: bridging biology, chemistry, and business. PMID- 18510380 TI - The effects of immunosuppression on regulatory CD4(+)CD25(+) T cells: impact on immunosuppression selection in transplantation. AB - During immune response and T-cell activation, both effector T cells and regulatory T(T(reg)) cells are activated and regulated simultaneously by both positive and negative pathways. CD4(+)CD25(+) T(reg) cells play a critical role in immune tolerance to self antigens as well as to allografts in some transplant settings. Effective immunosuppressive regimens significantly reduced the incidence of acute allograft rejection in patients following organ transplantation. However, the impact of immunosuppressive treatment on the potential induction of transplant tolerance has not been well determined. In this review we summarize the effects of immunosuppressive reagents on CD4(+)CD25(+) T(reg) cells in order to bring attention to this issue, which may affect the choice of immunosuppressive regimen in the clinical setting. PMID- 18510382 TI - Lucica MI urinary myoinositol kit: a new diagnostic test for diabetes mellitus and glucose intolerance. AB - Diabetes mellitus is a growing healthcare problem internationally, and poses a major burden from both a individual and societal perspective. Diabetes causes potentially life-threatening complications that are preventable if the disease is detected early and appropriate interventions are put in place. Early detection is therefore imperative for preventing diabetes-related morbidity and mortality. Current methods of detection, including the oral glucose tolerance test (OGTT), and measures of fasting plasma glucose, glycated hemoglobin (HbA(1c)), or glycated albumin, can be time-consuming and uncomfortable for patients. Myoinositol can be measured in urine and has been found to be elevated in patients with diabetes and glucose intolerance; it has thus proven useful as a marker for the early detection of these conditions. Lucica MI is a diagnostic kit for the measurement of urinary myoinositol; it is used to detect glucose intolerance and diabetes mellitus at an early stage in disease progression. The test is based on an enzymatic method that uses liquid reagents requiring no preparation. Clinical trial results demonstrate that the test could be used to detect not only diabetes mellitus, but also to distinguish impaired fasting glucose and impaired glucose tolerance from normal glucose tolerance. PMID- 18510383 TI - From the world literature. PMID- 18510384 TI - Let's talk.. PMID- 18510385 TI - Report to the membership...AAMI's new strategic plan charts future, expands member value. PMID- 18510387 TI - The technical iconoclast roundtable... thinking outside the box. PMID- 18510390 TI - Shop organization. PMID- 18510391 TI - Biomed roundup. PMID- 18510399 TI - Modernizing communications in healthcare: case studies highlight promise, pitfalls. PMID- 18510404 TI - A view from the corner command post. PMID- 18510405 TI - Speak up: a guide to improving your public speaking skills. PMID- 18510407 TI - Motivating challenging employees. PMID- 18510408 TI - Biomedical device interfacing to clinical information systems: a primer. AB - I am pleased that we get to take advantage of Bridget Moorman's background, experience, and perspective in this installment of IT World. One of the most nerve-racking tasks we run into these days is getting disparate medical devices to talk to each other over a network. This is especially so if the device you're trying to communicate with doesn't support network connectivity. Bridget shares her experience here not only with a great high-level view of network interfacing, but also with references to dig into all the grim details. She shows us a lot of facets to consider when assembling such a network. You've got to convert to hit the ramp then translate and aggregate before gaining access to the clinical information system cloud. If that doesn't make sense, read on! -Jeff Kabachinski, IT World columnist. PMID- 18510409 TI - Confronting the network challenge. PMID- 18510410 TI - Using human factors engineering principles to strengthen technology management. PMID- 18510411 TI - Getting up to speed on laser safety. PMID- 18510412 TI - Heart lung machines. PMID- 18510413 TI - Asset management: securing the most vulnerable assets. PMID- 18510414 TI - The "other" environment of care management plans. PMID- 18510415 TI - Laying the foundation for preventing device-related patient safety problems. PMID- 18510416 TI - The challenge of the modern sterile processing department. PMID- 18510417 TI - Purchasing controls: best practice guidelines. PMID- 18510418 TI - Guidance expands to include home hemodialysis. PMID- 18510419 TI - Using high-temperature formaldehyde sterilization as a model for studying gaseous sterilization. AB - This study uses the high-temperature formaldehyde sterilization system provided by the Harvey Chemiclave, manufactured by Barnstead Thermolyne Corporation (Dubuque, IA), as a model to investigate certain phenomena associated with gaseous chemical sterilization systems. Although formaldehyde sterilization presents some unique and complex system attributes, the current studies provide helpful insights into general sterilization methods by chemicals in the gaseous state. Both population recovery and fraction negative (FN) techniques were used to assay surviving populations from biological indicators of the organism Geobacillus stearothermophilus following exposure to incremental Chemiclave cycles. Models 5500 and 6000 of the Barnstead/Thermolyne Chemiclave were used in the study. Reusable instruments such as scalers, explorers, and various hinged pieces were tested in minimum versus maximum load studies. Population recovery study results demonstrated that lethality rates increase with time throughout the Chemiclave sterilization process and that there are significant variations in lethality according to load location. The population recovery data in conjunction with the FN studies and temperature data confirm that one-half the full-cycle time is not a good estimator of one-half the full-cycle lethality because lethality curves are concave downward and lethality varies by load location. This conclusion can also be applied to other types of gaseous, chemical sterilization such as ethylene oxide. The work outlined in this study was a result of investigations into the parameters affecting formaldehyde chemical vapor sterilization with the Harvey Chemiclave sterilizer. During these studies, it became apparent that results clearly depicted the effects of continued acceleration of the rate of microbial lethality, as well as variations in delivered lethality as a function of position in the sterilizer load. This publication focuses on these observations because they are important considerations for understanding general concepts of sterilization efficacy in process applications. Erroneous conclusions can be drawn when one evaluates sterilization without a thorough understanding of affecting variables. PMID- 18510420 TI - Damaged equipment: who should pay. PMID- 18510421 TI - Quantum dots for tumor-targeted drug delivery and cell imaging. PMID- 18510424 TI - Antibody-functionalized nano test tubes target breast cancer cells. AB - AIM: To develop nano test tubes that will deliver a biomedical payload to a specific cell type. METHODS: The template-synthesis method was used to prepare silica nano test tubes. An antibody that is specific for breast cancer cells was attached to the outer tube surfaces. A fluorophore was attached to the inner surfaces of the nano test tubes. The tubes were incubated with the breast cancer cells and the extent of attachment to the cell surfaces was investigated by fluorescence microscopy. RESULTS: Tubes modified on their outer surfaces with the target antibody showed enhanced attachment to breast-cancer cells, relative to tubes modified on their outer surfaces with a species and isotype-matched control antibody. CONCLUSIONS: This work is a first step toward demonstrating that nano test tubes can be used as cell-specific delivery vehicles. PMID- 18510425 TI - Detection of HIV-1 p24 Gag in plasma by a nanoparticle-based bio-barcode amplification method. AB - BACKGROUND: Detection of HIV-1 in patients is limited by the sensitivity and selectivity of available tests. The nanotechnology-based bio-barcode amplification method offers an innovative approach to detect specific HIV-1 antigens from diverse HIV-1 subtypes. We evaluated the efficacy of this protein detection method in detecting HIV-1 in men enrolled in the Chicago component of the Multicenter AIDS Cohort Study (MACS). METHODS: The method relies on magnetic microparticles with antibodies that specifically bind the HIV-1 p24 Gag protein and nanoparticles that are encoded with DNA and antibodies that can sandwich the target protein captured by the microparticle-bound antibodies. The aggregate sandwich structures are magnetically separated from solution, and treated to remove the conjugated barcode DNA. The DNA barcodes (hundreds per target) were identified by a nanoparticle-based detection method that does not rely on PCR. RESULTS: Of 112 plasma samples from HIV-1-infected subjects, 111 were positive for HIV-1 p24 Gag protein (range: 0.11-71.5 ng/ml of plasma) by the bio-barcode amplification method. HIV-1 p24 Gag protein was detected in only 23 out of 112 men by the conventional ELISA. A total of 34 uninfected subjects were negative by both tests. Thus, the specificity of the bio-barcode-amplification method was 100% and the sensitivity 99%. The bio-barcode-amplification method detected HIV-1 p24 Gag protein in plasma from all study subjects with less than 200 CD4(+) T cells/microl of plasma (100%) and 19 out of 20 (95%) HIV-1-infected men who had less than 50 copies/ml of plasma of HIV-1 RNA. In a separate group of 60 diverse international isolates, representative of clades A, B, C and D and circulating recombinant forms CRF01_AE and CRF02_AG, the bio-barcode-amplification method identified the presence of virus correctly. CONCLUSIONS: The bio-barcode amplification method was superior to the conventional ELISA assay for the detection of HIV-1 p24 Gag protein in plasma with a breadth of coverage for diverse HIV-1 subtypes. Because the bio-barcode-amplification method does not require enzymatic amplification, this method could be translated into a robust point-of-care test. PMID- 18510426 TI - Studies on biodegradable polymeric nanoparticles of risperidone: in vitro and in vivo evaluation. AB - AIM: The aim of this work was to develop extended-release risperidone nanoparticles for parenteral delivery (intravenous) and to reduce the dose dependent extrapyramidal side effects of risperidone. METHODS: Polymeric nanoparticles containing risperidone made of poly (epsilon-caprolactone) were designed by the nanoprecipitation method using polymeric stabilizers (poloxamers). The in vivo efficacy of prepared formulations and the risperidone solution was studied by administering them intravenously to apomorphine-treated mice. Extrapyramidal side effects of the risperidone and its formulations were also studied. RESULTS: The particle size of the prepared nanoparticles ranged between 99 and 304 nm. Approximately 78-85% drug-encapsulation efficiency was achieved when risperidone was loaded at 1.7-4.1% by weight of the polymer. During in vivo studies, prepared risperidone-containing formulations showed a significant prolonged antipsychotic effect than that of risperidone solution, also having less extrapyramidal side effects. CONCLUSION: The prolonged effect of risperidone was obtained from the nanoparticles of risperidone administered by the intravenous route and this may improve the treatment of psychotic disorders by dose reduction. PMID- 18510427 TI - A carbon-wrapped nanoscaled thermometer for temperature control in biological environments. AB - AIMS: A carbon-wrapped nanoscaled thermometer for a contactless temperature control in biological systems on the cellular level is presented. MATERIALS & METHODS: The thermometer is based on multiwalled carbon nanotubes (MWCNTs) filled with materials with strongly temperature-dependent nuclear magnetic resonance (NMR) parameters. The NMR frequency shift and relaxation time were measured in cuprous-iodide-filled CNTs at different temperatures. RESULTS: The experimental data indicate a pronounced temperature dependence of the NMR parameters, thereby realizing the nanoscaled thermometer. CONCLUSION: This study is a proof-of concept that the functionalized CNTs can be used as a contactless thermometer in biomedical applications. PMID- 18510428 TI - Antibacterial nanomedicine. AB - Recent advances in the field of nanotechnology led several groups to recognize the promise of recruiting nanomaterials to the ongoing battle against pathogenic bacteria. A large battery of newly discovered and developed nanomaterials has been accumulating during the last decade, therefore, it could be anticipated that it should only be a matter of time until such preliminary nanomedicine applications are presented. We review some of these pioneering studies in which nanomaterials have been evaluated as potential therapeutics, antiseptics or disinfectants. These studies can be divided roughly into two groups. The first are studies of antibacterial nanomedicines that are based solely on synthetic (artificial) materials. The second group comprises studies of antibacterial nanomaterials that are based on biological substances used in their natural or in a modified form. We will discuss the physicochemical and antibacterial highlights of each material and present the future perspectives of this emerging field. PMID- 18510430 TI - Intracellular delivery of nanoparticles via the HIV-1 tat peptide. AB - Functionalized nanoparticles are heralded as part of the future with regards to targeted cell and nuclear delivery. However, direct intracellular and intranuclear delivery has, until recently, been difficult to achieve owing to the impermeable nature of the plasma and nuclear membranes. During the past 15 years, a range of peptides, termed cell-penetrating peptides (CPPs), which have the ability to translocate into living cells, have been discovered. Thus, in more recent years, the combination of CPPs with nanoparticles, enabling CPP-mediated cell delivery, has opened up many avenues of research. This review discusses the use of various CPPs, focusing on tat peptide, to functionalize nanoparticles and the possible move from the laboratory to the clinic. PMID- 18510429 TI - Design opportunities for actively targeted nanoparticle vaccines. AB - Vaccines for many infectious diseases are poorly developed or simply unavailable. There are significant technological and practical design issues that contribute to this problem; thus, a solution to the vaccine problem will require a systematic approach to test the multiple variables that are required to address each of the design challenges. Nanoparticle technology is an attractive methodology for optimizing vaccine development because design variables can be tested individually or in combination. The biology of individual components that constitute an effective vaccine is often well understood and may be integrated into particle design, affording optimal immune responses to specific pathogens. Here, we review technological variables and design parameters associated with creating modular nanoparticle vaccine systems that can be used as vectors to protect against disease. Variables, such as the material and size of the core matrix, surface modification for attaching targeting ligands and routes of administration, are discussed. Optimization of these variables is important for the development of nanoparticle-based vaccine systems against infectious diseases and cancer. PMID- 18510431 TI - Micro/nanostructured polymeric systems for biomedical and pharmaceutical applications. AB - This review provides an outline of the polymeric micro/nanostructured advanced systems that are suited for the controlled and targeted administration of, specifically, nonconventional drugs. The contribution of new trends in drug delivery technology is focused on two major parts, dealing with brief surveys of: the biodegradable/bioerodible polymeric systems used in the formulation of micro/nanoparticles and techniques used in the preparation of micro/nanoparticles for their biomedical application in cancer treatment specifically, in inflammation pathologies, as oxygen carriers (blood substitutes) and in tissue engineering practice. A small discussion of the future perspectives of the described systems is also given. PMID- 18510435 TI - Calcium is involved in formation of high molecular weight adiponectin. AB - BACKGROUND: Adiponectin, an adipocyte-specific secretory protein, is known to circulate as different isoforms in the blood stream. METHODS: Using sucrose gradients and Western blotting on nondenaturing gels, adiponectin isoforms were examined in human serum, plasma, adipose tissue, and cells. The medium from human adipose tissue and human and mouse adipocytes were also examined for changes in isoform formation upon treatment with EGTA. RESULTS: Comparison of adiponectin complexes revealed distinct differences in distribution of high molecular weight (HMW) forms between human serum and plasma, with an apparent difference in molecular weight. Variation in molecular weight suggested a probable dissociation of the HMW isoforms in the presence of EDTA in the plasma. Examination of human serum samples treated with EDTA or EGTA showed a partial dissociation of the HMW isoform, while the addition of excess calcium, but not magnesium, to human plasma resulted in partial restoration of HMW adiponectin. When human adipose tissue secreted adiponectin was treated with EGTA, there was a decrease in the HMW isoform by 61% (+/- 1.89%) and a corresponding increase in low molecular weight (LMW) and middle molecular weight (MMW) isoforms, compared to untreated samples. Analysis of mouse and human adipocytes also showed a reduction in HMW isoforms with a corresponding increase in MMW and LMW isoforms upon treatment with EGTA. The Simpson-Golabi-Behmel syndrome (SGBS) human adipocyte cell line, which primarily synthesizes LMW isoforms, produced increasing amounts of HMW adiponectin upon treatment with calcium in a dose-dependent manner. CONCLUSION: These data indicate that calcium promotes the formation of HMW adiponectin, and calcium sequestration decreases HMW adiponectin. Because of the importance of HMW adiponectin in insulin sensitivity, these data demonstrate the importance of assay conditions and sample preparation in the measurement of adiponectin isoforms. PMID- 18510434 TI - Physiological, pharmacological, and nutritional regulation of circulating adiponectin concentrations in humans. AB - Adiponectin is an adipocyte hormone that links visceral adiposity with insulin resistance and atherosclerosis. It is unique among adipocyte-derived hormones in that its circulating concentrations are inversely proportional to adiposity, and low adiponectin concentrations predict the development of type 2 diabetes and cardiovascular disease. Consequently, in the decade since its discovery, adiponectin has generated immense interest as a potential therapeutic target for the metabolic syndrome and diabetes. This review summarizes current research regarding the regulation of circulating adiponectin concentrations by physiological, pharmacological, and nutritional factors, with an emphasis on human studies. In humans, plasma adiponectin concentrations are influenced by age and gender, and are inversely proportional to visceral adiposity. In vitro studies suggest that adiponectin production may be determined primarily by adipocyte size and insulin sensitivity, with larger, insulin-resistant adipocytes producing less adiponectin. While adiponectin concentrations are unchanged after meal ingestion, they are increased by significant weight loss, such as after bariatric surgery. In addition, adiponectin production is inhibited by a number of hormones, including testosterone, prolactin, glucocorticoids and growth hormone, and by inflammation and oxidative stress in adipose tissue. Smoking decreases, while moderate alcohol consumption increases, circulating adiponectin concentrations. Dietary fatty acid composition in rodents influences adiponectin production via ligand-activated nuclear receptors (PPARs); however, current evidence in humans is equivocal. In addition to PPAR agonists (such as thiazolidinediones and fibrates), a number of pharmacological agents (angiotensin receptor type 1 blockers, ACE inhibitors, and cannabinoid receptor antagonists) used in treatment of the metabolic syndrome also increase adiponectin concentrations in humans. PMID- 18510436 TI - Obesity in Jordan: prevalence, associated factors, comorbidities, and change in prevalence over ten years. AB - OBJECTIVES: To determine the prevalence of obesity in northern Jordan, identify its associated factors, assess its association with selected comorbidities, and determine how the prevalence of obesity has changed in Jordan over 10 years. METHODS: A total of 1121 participants aged 25 years and above were randomly selected. Sociodemographic characteristics as well as information on selected metabolic disorders and their potential risk factors were obtained. Anthropometric and biochemical characteristics were measured. Obesity was defined based on body mass index (BMI), waist circumference, and waist-to-hip ratio. RESULTS: The age-standardized prevalence of obesity in northern Jordan was 28.1% (95% CI: 23.4, 32.8) for men and 53.1% (95% CI: 49.3, 57.0) for women. Irrespective of age or measure used, women always had a considerably higher prevalence of obesity than men. The prevalence of obesity varied greatly with age, generally increasing, irrespective of the measurement used. There has been a significant increase in the prevalence of obesity over a period of ten years for both men and women aged 60 years and above only. When important variables were taken into account in logistic regression analyses, obesity was significantly associated with increased odds of having all studied metabolic abnormalities. Female gender, increase in age, being married, former smoker or nonsmoker, and fewer than 12 years of education were significantly associated with increased odds of BMI-defined obesity and high waist circumference. CONCLUSIONS: This study demonstrated alarming rates of obesity and of its associated comorbidities among Jordanians, especially among women. PMID- 18510437 TI - Effects of exercise and serum uric acid on the metabolic syndrome for Japanese workers. AB - BACKGROUND: The effects of exercise and serum uric acid on the metabolic syndrome (MetS) were evaluated. METHODS: The study subjects were 1792 Japanese male workers aged 21 to 60 years old who were employees of a manufacturing company of electrical products. The standard Japanese criteria for the diagnosis of MetS were used. The presence of obesity is an indispensable factor, with any two or more of the following criteria: (1) a high serum level of triglyceride and/or low serum level of HDL cholesterol; (2) high blood pressure; (3) high fasting plasma glucose concentration. The body mass index (BMI) was used as an obesity-related overweight index; BMI >or=25 kg/m(2) was defined as overweight. Age, smoking habit and frequency of drinking were used for the adjustment of significance. RESULTS: The odds ratios (95% confidence intervals) of serum uric acid >or=7 mg/dL, and no history of exercise for MetS were 3.2 (2.2, 4.6) and 1.8 (1.1, 3.0), respectively. The authors also categorized walking by the number of steps per day and by the time in minute per day. CONCLUSIONS: Although subjects with no habitual exercise have risk of MetS, the situation of physical activity should precisely be evaluated. PMID- 18510438 TI - The attentional blink modulates activity in the early visual cortex. AB - The attentional blink (AB) documents a particularly strong case of visual attentional competition, in which subjects' ability to identify a second target (T2) is significantly impaired when it is presented with a short SOA after a first target (T1). We used functional magnetic resonance imaging to investigate the impact of the AB on visual activity in individually defined retinotopic representations of the target stimuli. Our results show reduction of neural response in V3 and marginally in V2 and V1, paralleling the behavioral AB effect. Reduction of visual activity was accompanied by reduced neural response in the inferior parietal cortex. This indicates that attentional competition modulates activity in higher-order parietal regions and the early visual cortex, providing a plausible neural basis of the behavioral AB effect. PMID- 18510439 TI - Functional representation of living and nonliving domains across the cerebral hemispheres: a combined event-related potential/transcranial magnetic stimulation study. AB - Transcranial magnetic stimulation (TMS) over the left hemisphere has been shown to disrupt semantic processing but, to date, there has been no direct demonstration of the electrophysiological correlates of this interference. To gain insight into the neural basis of semantic systems, and in particular, study the temporal and functional organization of object categorization processing, we combined repetitive TMS (rTMS) and ERPs. Healthy volunteers performed a picture word matching task in which Snodgrass drawings of natural (e.g., animal) and artifactual (e.g., tool) categories were associated with a word. When short trains of high-frequency rTMS were applied over Wernicke's area (in the region of the CP5 electrode) immediately before the stimulus onset, we observed delayed response times to artifactual items, and thus, an increased dissociation between natural and artifactual domains. This behavioral effect had a direct ERP correlate. In the response period, the stimuli from the natural domain elicited a significant larger late positivity complex than those from the artifactual domain. These differences were significant over the centro-parietal region of the right hemisphere. These findings demonstrate that rTMS interferes with post perceptual categorization processing of natural and artifactual stimuli that involve separate subsystems in distinct cortical areas. PMID- 18510440 TI - Time course of early audiovisual interactions during speech and nonspeech central auditory processing: a magnetoencephalography study. AB - Cross-modal fusion phenomena suggest specific interactions of auditory and visual sensory information both within the speech and nonspeech domains. Using whole head magnetoencephalography, this study recorded M50 and M100 fields evoked by ambiguous acoustic stimuli that were visually disambiguated to perceived /ta/ or /pa/ syllables. As in natural speech, visual motion onset preceded the acoustic signal by 150 msec. Control conditions included visual and acoustic nonspeech signals as well as visual-only and acoustic-only stimuli. (a) Both speech and nonspeech motion yielded a consistent attenuation of the auditory M50 field, suggesting a visually induced "preparatory baseline shift" at the level of the auditory cortex. (b) Within the temporal domain of the auditory M100 field, visual speech and nonspeech motion gave rise to different response patterns (nonspeech: M100 attenuation; visual /pa/: left-hemisphere M100 enhancement; /ta/: no effect). (c) These interactions could be further decomposed using a six dipole model. One of these three pairs of dipoles (V270) was fitted to motion induced activity at a latency of 270 msec after motion onset, that is, the time domain of the auditory M100 field, and could be attributed to the posterior insula. This dipole source responded to nonspeech motion and visual /pa/, but was found suppressed in the case of visual /ta/. Such a nonlinear interaction might reflect the operation of a binary distinction between the marked phonological feature "labial" versus its underspecified competitor "coronal." Thus, visual processing seems to be shaped by linguistic data structures even prior to its fusion with auditory information channel. PMID- 18510441 TI - Difficulty of discrimination modulates attentional capture by regulating attentional focus. AB - Attentional capture for distractors is enhanced by increasing the difficulty of discrimination between the standard and the target in the three-stimulus oddball paradigm. In this study, we investigated the cognitive mechanism of this modulation of attentional capture. Event-related brain potentials were recorded from participants while they performed a visual three-stimulus oddball paradigm (frequent standard, rare target, and rare distractor). The discrimination difficulty between standard and target was manipulated in the central location. Distractor stimuli were presented in the central or surrounding locations. The P3a component was elicited by distractor stimuli and was used as a measure of attentional capture. The results revealed that discrimination difficulty had opposite effects on the P3a response between central and surrounding locations. With an increase in the difficulty of discrimination, the P3a response was enhanced when distractor stimuli were presented in the central location. In contrast, the P3a response was reduced when distractor stimuli were presented in a surrounding location. This finding suggests that spatial attention was focused by the difficulty of discrimination, and deviant processing was increased within its focus but decreased outside its focus. Therefore, attentional capture for deviant distractors is modulated by top-down controlled attentional focus. PMID- 18510442 TI - Saccades to a remembered location elicit spatially specific activation in human retinotopic visual cortex. AB - The possible impact upon human visual cortex from saccades to remembered target locations was investigated using functional magnetic resonance imaging (fMRI). A specific location in the upper-right or upper-left visual quadrant served as the saccadic target. After a delay of 2,400 msec, an auditory signal indicated whether to execute a saccade to that location (go trial) or to cancel the saccade and remain centrally fixated (no-go). Group fMRI analysis revealed activation specific to the remembered target location for executed saccades, in the contralateral lingual gyrus. No-go trials produced similar, albeit significantly reduced, effects. Individual retinotopic mapping confirmed that on go trials, quadrant-specific activations arose in those parts of ventral V1, V2, and V3 that coded the target location for the saccade, whereas on no-go trials, only the corresponding parts of V2 and V3 were significantly activated. These results indicate that a spatial-motor saccadic task (i.e., making an eye movement to a remembered location) is sufficient to activate retinotopic visual cortex spatially corresponding to the target location, and that this activation is also present (though reduced) when no saccade is executed. We discuss the implications of finding that saccades to remembered locations can affect early visual cortex, not just those structures conventionally associated with eye movements, in relation to recent ideas about attention, spatial working memory, and the notion that recently activated representations can be "refreshed" when needed. PMID- 18510443 TI - Compressed scaling of abstract numerosity representations in adult humans and monkeys. AB - There is general agreement that nonverbal animals and humans endowed with language possess an evolutionary precursor system for representing and comparing numerical values. However, whether nonverbal numerical representations in human and nonhuman primates are quantitatively similar and whether linear or logarithmic coding underlies such magnitude judgments in both species remain elusive. To resolve these issues, we tested the numerical discrimination performance of human subjects and two rhesus monkeys (Macaca mulatta) in an identical delayed match-to-numerosity task for a broad range of numerosities from 1 to 30. The results demonstrate a noisy nonverbal estimation system obeying Weber's Law in both species. With average Weber fractions in the range of 0.51 and 0.60, nonverbal numerosity discriminations in humans and monkeys showed similar precision. Moreover, the detailed analysis of the performance distributions exhibited nonlinearly compressed numerosity representations in both primate species. However, the difference between linear and logarithmic scaling was less pronounced in humans. This may indicate a gradual transformation of a logarithmic to linear magnitude scale in human adults as the result of a cultural transformation process during the course of mathematical education. PMID- 18510444 TI - Oscillatory activity and phase-amplitude coupling in the human medial frontal cortex during decision making. AB - Electroencephalogram oscillations recorded both within and over the medial frontal cortex have been linked to a range of cognitive functions, including positive and negative feedback processing. Medial frontal oscillatory characteristics during decision making remain largely unknown. Here, we examined oscillatory activity of the human medial frontal cortex recorded while subjects played a competitive decision-making game. Distinct patterns of power and cross trial phase coherence in multiple frequency bands were observed during different decision-related processes (e.g., feedback anticipation vs. feedback processing). Decision and feedback processing were accompanied by a broadband increase in cross-trial phase coherence at around 220 msec, and dynamic fluctuations in power. Feedback anticipation was accompanied by a shift in the power spectrum from relatively lower (delta and theta) to higher (alpha and beta) power. Power and cross-trial phase coherence were greater following losses compared to wins in theta, alpha, and beta frequency bands, but were greater following wins compared to losses in the delta band. Finally, we found that oscillation power in alpha and beta frequency bands were synchronized with the phase of delta and theta oscillations ("phase-amplitude coupling"). This synchronization differed between losses and wins, suggesting that phase-amplitude coupling might reflect a mechanism of feedback valence coding in the medial frontal cortex. Our findings link medial frontal oscillations to decision making, with relations among activity in different frequency bands suggesting a phase-utilizing coding of feedback valence information. PMID- 18510445 TI - An event-related potential study on changes of violation and error responses during morphosyntactic learning. AB - Based on recent findings showing electrophysiological changes in adult language learners after relatively short periods of training, we hypothesized that adult Dutch learners of German would show responses to German gender and adjective declension violations after brief instruction. Adjective declension in German differs from previously studied morphosyntactic regularities in that the required suffixes depend not only on the syntactic case, gender, and number features to be expressed, but also on whether or not these features are already expressed on linearly preceding elements in the noun phrase. Violation phrases and matched controls were presented over three test phases (pretest and training on the first day, and a posttest one week later). During the pretest, no electrophysiological differences were observed between violation and control conditions, and participants' classification performance was near chance. During the training and posttest phases, classification improved, and there was a P600-like violation response to declension but not gender violations. An error-related response during training was associated with improvement in grammatical discrimination from pretest to posttest. The results show that rapid changes in neuronal responses can be observed in adult learners of a complex morphosyntactic rule, and also that error-related electrophysiological responses may relate to grammar acquisition. PMID- 18510446 TI - Attentional selection of multiple goal positions before rapid hand movement sequences: an event-related potential study. AB - A dot-probe paradigm was used to provide physiological evidence for the parallel selection of multiple movement goals before rapid hand movement sequences. Participants executed a sequence of manual pointing movements to two out of three possible goal positions. During movement preparation, a task-irrelevant visual transient (a dot probe) was flashed either at one of both movement goals, or at the third, movement-irrelevant location. The results revealed that the N1 component induced by the presentation of the dot was enhanced if the dot was flashed at one of the movement goals, indicating that both target positions were attended before the initialization of the movement sequence. A second experiment showed that movement-irrelevant locations between the movement goals were not attended, suggesting that attention splits into spatially distinct foci. PMID- 18510447 TI - Structural and metabolic correlates of episodic memory in relation to the depth of encoding in normal aging. AB - This study set out to establish the relationship between changes in episodic memory retrieval in normal aging on the one hand and gray matter volume and (18)FDG uptake on the other. Structural MRI, resting-state (18)FDG-PET, and an episodic memory task manipulating the depth of encoding and the retention interval were administered to 46 healthy subjects divided into three groups according to their age (young, middle-aged, and elderly adults). Memory decline was found not to be linear in the course of normal aging: Whatever the retention interval, the retrieval of shallowly encoded words was impaired in both the middle-aged and the elderly, whereas the retrieval of deeply encoded words only declined in the elderly. In middle-aged and elderly subjects, the reduced performance in the shallow encoding condition was mainly related to posterior mediotemporal volume and metabolism. By contrast, the impaired retrieval of deeply encoded words in the elderly group was mainly related to frontal and parietal regions, suggesting the adoption of inefficient strategic processes. PMID- 18510448 TI - Development of spatial and verbal working memory capacity in the human brain. AB - A core aspect of working memory (WM) is the capacity to maintain goal-relevant information in mind, but little is known about how this capacity develops in the human brain. We compared brain activation, via fMRI, between children (ages 7-12 years) and adults (ages 20-29 years) performing tests of verbal and spatial WM with varying amounts (loads) of information to be maintained in WM. Children made disproportionately more errors than adults as WM load increased. Children and adults exhibited similar hemispheric asymmetry in activation, greater on the right for spatial WM and on the left for verbal WM. Children, however, failed to exhibit the same degree of increasing activation across WM loads as was exhibited by adults in multiple frontal and parietal cortical regions. Thus, children exhibited adult-like hemispheric specialization, but appeared immature in their ability to marshal the neural resources necessary to maintain large amounts of verbal or spatial information in WM. PMID- 18510449 TI - Distinctive neural mechanisms supporting visual object individuation and identification. AB - Many everyday activities, such as driving on a busy street, require the encoding of distinctive visual objects from crowded scenes. Given resource limitations of our visual system, one solution to this difficult and challenging task is to first select individual objects from a crowded scene (object individuation) and then encode their details (object identification). Using functional magnetic resonance imaging, two distinctive brain mechanisms were recently identified that support these two stages of visual object processing. While the inferior intraparietal sulcus (IPS) selects a fixed number of about four objects via their spatial locations, the superior IPS and the lateral occipital complex (LOC) encode the features of a subset of the selected objects in great detail (object shapes in this case). Thus, the inferior IPS individuates visual objects from a crowded display and the superior IPS and higher visual areas participate in subsequent object identification. Consistent with the prediction of this theory, even when only object shape identity but not its location is task relevant, this study shows that object individuation in the inferior IPS treats four identical objects similarly as four objects that are all different, whereas object shape identification in the superior IPS and the LOC treat four identical objects as a single unique object. These results provide independent confirmation supporting the dissociation between visual object individuation and identification in the brain. PMID- 18510450 TI - Alternate task inhibits single-neuron category-selective responses in the human hippocampus while preserving selectivity in the amygdala. AB - One fifth of neurons in the medial-temporal lobe of human epilepsy patients respond selectively to categories of images, such as faces or cars. Here we show that responses of hippocampal neurons are rapidly modified as subjects alternate (over 60 sec) between two tasks (1) identifying images from a category, or (2) playing a simple video game superimposed on the same images. Category-selective responses, present when a subject identifies categories, are eliminated when the subject shifts to playing the game for 87% of category-selective hippocampal neurons. By contrast, responses in the amygdala are present during both tasks for 72% of category-selective amygdalar neurons. These results suggest that attention to images is required to evoke selective responses from single neurons in the hippocampus, but is not required by neurons in the amygdala. PMID- 18510451 TI - Are vowels and consonants processed differently? Event-related potential evidence with a delayed letter paradigm. AB - To investigate the neural bases of consonant and vowel processing, event-related potentials (ERPs) were recorded while participants read words and pseudowords in a lexical decision task. The stimuli were displayed in three different conditions: (i) simultaneous presentation of all letters (baseline condition); (ii) presentation of all letters, except that two internal consonants were delayed for 50 msec (consonants-delayed condition); and (iii) presentation of all letters, except that two internal vowels were delayed for 50 msec (vowels-delayed condition). The behavioral results showed that, for words, response times in the consonants-delayed condition were longer than in the vowels-delayed condition, which, in turn, were longer than in the baseline condition. The ERPs showed that, starting as early as 150 msec, words in the consonants-delayed condition produced a larger negativity than words in vowels-delayed condition. In addition, there were peak latency differences and amplitude differences in the P150, N250, P325, and N400 components between the baseline and the two letter-delayed conditions. We examine the implications of these findings for models of visual-word recognition and reading. PMID- 18510452 TI - The common neural basis of autobiographical memory, prospection, navigation, theory of mind, and the default mode: a quantitative meta-analysis. AB - A core brain network has been proposed to underlie a number of different processes, including remembering, prospection, navigation, and theory of mind [Buckner, R. L., & Carroll, D. C. Self-projection and the brain. Trends in Cognitive Sciences, 11, 49-57, 2007]. This purported network-medial prefrontal, medial-temporal, and medial and lateral parietal regions-is similar to that observed during default-mode processing and has been argued to represent self projection [Buckner, R. L., & Carroll, D. C. Self-projection and the brain. Trends in Cognitive Sciences, 11, 49-57, 2007] or scene-construction [Hassabis, D., & Maguire, E. A. Deconstructing episodic memory with construction. Trends in Cognitive Sciences, 11, 299-306, 2007]. To date, no systematic and quantitative demonstration of evidence for this common network has been presented. Using the activation likelihood estimation (ALE) approach, we conducted four separate quantitative meta-analyses of neuroimaging studies on: (a) autobiographical memory, (b) navigation, (c) theory of mind, and (d) default mode. A conjunction analysis between these domains demonstrated a high degree of correspondence. We compared these findings to a separate ALE analysis of prospection studies and found additional correspondence. Across all domains, and consistent with the proposed network, correspondence was found within the medial-temporal lobe, precuneus, posterior cingulate, retrosplenial cortex, and the temporo-parietal junction. Additionally, this study revealed that the core network extends to lateral prefrontal and occipital cortices. Autobiographical memory, prospection, theory of mind, and default mode demonstrated further reliable involvement of the medial prefrontal cortex and lateral temporal cortices. Autobiographical memory and theory of mind, previously studied as distinct, exhibited extensive functional overlap. These findings represent quantitative evidence for a core network underlying a variety of cognitive domains. PMID- 18510453 TI - In vitro electrophysiological drug testing using human embryonic stem cell derived cardiomyocytes. AB - Pro-arrhythmia (development of cardiac arrhythmias as a pharmacological side effect) has become the single most common cause of the withdrawal or restrictions of previously marketed drugs. The development of new medications, free from these side effects, is hampered by the lack of an in vitro assay for human cardiac tissue. We hypothesized that human embryonic stem cell-derived cardiomyocytes (hESC-CMs) assessed with a combination of single cell electrophysiology and microelectrode array (MEA) mapping can serve as a novel model for electrophysiological drug screening. Current-clamp studies revealed that E-4031 and Sotalol (IKr blockers) significantly increased hESC-CM's action potential duration and also induced after-depolarizations (the in vitro correlates of increased arrhythmogenic potential). Multicellular aggregates of hESC-CMs were then analyzed with the MEA technique. Application of class I (Quinidine, Procaineamide) and class III (Sotalol) antiarrhythmic agents, E-4031, and Cisapride (a noncardiogenic agent known to lengthen QT) resulted in dose dependent prolongation of the corrected field potential duration (cFPD). We next utilized the MEA technique to also assess pharmacological effects on conduction. Activation maps demonstrated significant conduction slowing following administration of Na channel blockers (Quinidine and Propafenone) and of the gap junction blocker (1-heptanol). While most attention has been focused on the prospects of using hESC-derived cardiomyocytes for regenerative medicine, this study highlights the possible utilization of these unique cells also for cardiac electrophysiological studies, drug screening, and target validation. PMID- 18510454 TI - Protective role of DC-SIGN (CD209) neck-region alleles with <5 repeat units in HIV-1 transmission. AB - To investigate the role that DC-SIGN neck-region length variation plays in HIV-1 transmission, we studied 530 HIV-1-positive and 341 HIV-1-negative individuals in China. The carrier frequency of a DC-SIGN allele with <5 repeat units in the neck region was 0.9% in HIV-1-positive and 3.8% in HIV-1-negative individuals (P=.007), an observation suggesting that this DC-SIGN variation plays a role in HIV-1 transmission. These naturally occurring DC-SIGN neck-region variants were significantly more frequent in the Chinese population than in the US population (P <.001) and in a worldwide population (P=.006). PMID- 18510455 TI - Risk factors for surgical site infection after low transverse cesarean section. AB - BACKGROUND: Independent risk factors for surgical site infection (SSI) after cesarean section have not been well documented, despite the large number of cesarean sections performed and the relatively common occurrence of SSI. OBJECTIVE: To determine independent risk factors for SSI after low transverse cesarean section. DESIGN: Retrospective case-control study. SETTING: Barnes Jewish Hospital, a 1,250-bed tertiary care hospital. PATIENTS: A total of 1,605 women who underwent low transverse cesarean section during the period from July 1999 to June 2001. METHODS: Using the International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes for SSI or wound complication and/or data on antibiotic use during the surgical hospitalization or at readmission to the hospital or emergency department, we identified potential cases of SSI in a cohort of patients who underwent a low transverse cesarean section. Cases of SSI were verified by chart review using the definitions from the Centers for Disease Control and Prevention's National Nosocomial Infections Surveillance System. Control patients without SSI or endomyometritis were randomly selected from the population of patients who underwent cesarean section. Independent risk factors for SSI were determined by logistic regression. RESULTS: SSIs were identified in 81 (5.0%) of 1,605 women who underwent low transverse cesarean section. Independent risk factors for SSI included development of subcutaneous hematoma after the procedure (adjusted odds ratio [aOR], 11.6 [95% confidence interval [CI], 4.1-33.2]), operation performed by the university teaching service (aOR, 2.7 [95% CI, 1.4-5.2]), and a higher body mass index at admission (aOR, 1.1 [95% CI, 1.0-1.1]). Cephalosporin therapy before or after the operation was associated with a significantly lower risk of SSI (aOR, 0.2 [95% CI, 0.1-0.5]). Use of staples for skin closure was associated with a marginally increased risk of SSI. CONCLUSIONS: These independent risk factors should be incorporated into approaches for the prevention and surveillance of SSI after surgery. PMID- 18510457 TI - Decreased rates of nosocomial endometritis and urinary tract infection after vaginal delivery in a French surveillance network, 1997-2003. AB - OBJECTIVES: To identify independent risk factors for endometritis and urinary tract infection (UTI) after vaginal delivery, and to monitor changes in nosocomial infection rates and derive benchmarks for prevention. DESIGN: Prospective study. METHODS: We analyzed routine surveillance data for all vaginal deliveries between January 1997 and December 2003 at 66 maternity units participating in the Mater Sud-Est surveillance network. Adjusted odds ratios for risk of endometritis or UTI were obtained using a logistic regression model. RESULTS: The overall incidence rates were 0.5% for endometritis and 0.3% for UTI. There was a significant decrease in the incidence and risk of endometritis but not of UTI during the 7-year period. Significant risk factors for endometritis were fever during labor, parity of 1, and instrumental delivery and/or manual removal of the placenta. Significant risk factors for UTI were urinary infection on admission, premature rupture of membranes (more than 12 hours before admission), blood loss of more than 800 mL, parity of 1, instrumental delivery, and receipt of more than 5 vaginal digital examinations. Each maternity unit received a poster showing graphs of the number of expected and observed cases of UTI and endometritis associated with vaginal deliveries, which enabled each maternity unit to determine their rank within the network and to initiate prevention programs. CONCLUSIONS: Although routine surveillance means additional work for maternity units, our results demonstrate the usefulness of regular targeted monitoring of risk factors and of the most common nosocomial infections in obstetrics. Most of the information needed for monitoring is already present in the patients' records. PMID- 18510458 TI - Time-series analysis of the impact of bed occupancy rates on the incidence of methicillin-resistant Staphylococcus aureus infection in overcrowded general wards. AB - OBJECTIVE: We investigated the impact of bed occupancy, particularly overcrowding, on the incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection in general ward settings. METHODS: We performed a time-series and mixed-model analysis of variance of monthly incidence of MRSA infection and corresponding bed occupancy rates, over 65 months, in the medicine and surgical wards within St. Luke's Hospital, a 900-bed tertiary care facility in Malta. RESULTS: In the medicine wards, significant periodic fluctuations in bed demand were evident during the study period, with peaks of occupancy greater than 120% during the winter months. Cross-correlation analysis between the rate of bed occupancy and the rate of MRSA infection displayed an oscillatory configuration, with a periodicity of 12, similar to the periodicity evident in the autocorrelation bed-occupancy pattern. Further statistical analysis by means of analysis of variance confirmed that the months with excessive overcrowding tended to coincide with a significant increase in the rate of MRSA infection, occurring after a lag of approximately 2 months. Identical analysis of equivalent data from the surgical wards also revealed significant fluctuation in the rate of bed occupancy; however, occupancy never exceeded 100%. No cross-correlational relationship with MRSA infection incidence was present. CONCLUSION: The study data suggest that, in our setting, simple fluctuations in the rate of bed occupancy did not have a direct impact on the incidence of MRSA infection as long as the rate of bed occupancy was within designated levels. Rather, it was episodes of significant overcrowding, with occupancy levels in excess of designated numbers, that triggered increases in infection incidence rates. PMID- 18510459 TI - Active surveillance to determine the impact of methicillin-resistant Staphylococcus aureus colonization on patients in intensive care units of a Veterans Affairs Medical Center. AB - OBJECTIVE: The impact of methicillin-resistant Staphylococcus aureus (MRSA) colonization on mortality has not been well characterized. We sought to describe the impact of MRSA colonization on patients admitted to intensive care units (ICUs) in the Birmingham Veterans Affairs Medical Center (VAMC). METHODS: We conducted a retrospective cohort study of ICU patients at the Birmingham VAMC during 2005 to evaluate the predictors of MRSA colonization and determine its effect on clinical outcomes. Surveillance cultures for MRSA were performed on admission to the ICU and weekly thereafter. Clinical findings, the incidence of MRSA infection, and mortality within 3 months after ICU admission were recorded. Predictors of mortality and S. aureus colonization were determined using multivariable models. RESULTS: S. aureus colonization was present in 97 (23.3%) of 416 patients screened, of whom 67 (16.1%) were colonized with methicillin susceptible S. aureus (MSSA) and 30 (7.2%) with MRSA. All-cause mortality at 3 months among MRSA-colonized patients was significantly greater than that among MSSA-colonized patients (46.7% vs 19.4%; P = .009). MRSA colonization was an independent predictor of death (adjusted odds ratio [OR], 3.7 [95% confidence interval [CI], 1.5-8.9]; P = .003) and onset of MRSA infection after hospital discharge (adjusted OR, 7.6 [95% CI, 2.48-23.2]; P < .001). Risk factors for MRSA colonization included recent antibiotic use (adjusted OR, 4.8 [95% CI, 1.9-12.2]; P = .001) and dialysis (adjusted OR, 18.9 [95% CI, 2.1-167.8]; P = .008). CONCLUSIONS: Among ICU patients, MRSA colonization is associated with subsequent MRSA infection and an all-cause mortality that is greater than that for MSSA colonization. Active surveillance for MRSA colonization may identify individuals at risk for these adverse outcomes. Prospective studies of outcomes in MRSA colonized patients may better define the role of programs for active MRSA surveillance. PMID- 18510460 TI - Highly effective regimen for decolonization of methicillin-resistant Staphylococcus aureus carriers. AB - OBJECTIVE: To evaluate the efficacy of a standardized regimen for decolonization of methicillin-resistant Staphylococcus aureus (MRSA) carriers and to identify factors influencing decolonization treatment failure. DESIGN: Prospective cohort study from January 2002 to April 2007, with a mean follow-up period of 36 months. SETTING: University hospital with 750 beds and 27,000 admissions/year. PATIENTS: Of 94 consecutive hospitalized patients with MRSA colonization or infection, 32 were excluded because of spontaneous loss of MRSA, contraindications, death, or refusal to participate. In 62 patients, decolonization treatment was completed. At least 6 body sites were screened for MRSA (including by use of rectal swabs) before the start of treatment. INTERVENTIONS: Standardized decolonization treatment consisted of mupirocin nasal ointment, chlorhexidine mouth rinse, and full-body wash with chlorhexidine soap for 5 days. Intestinal and urinary-tract colonization were treated with oral vancomycin and cotrimoxazole, respectively. Vaginal colonization was treated with povidone-iodine or, alternatively, with chlorhexidine ovula or octenidine solution. Other antibiotics were added to the regimen if treatment failed. Successful decolonization was considered to have been achieved if results were negative for 3 consecutive sets of cultures of more than 6 screening sites. RESULTS: The mean age (+/- standard deviation [SD]) age of the 62 patients was 66.2 +/- 19 years. The most frequent locations of MRSA colonization were the nose (42 patients [68%]), the throat (33 [53%]), perianal area (33 [53%]), rectum (36 [58%]), and inguinal area (30 [49%]). Decolonization was completed in 87% of patients after a mean (+/-SD) of 2.1 +/- 1.8 decolonization cycles (range, 1-10 cycles). Sixty-five percent of patients ultimately required peroral antibiotic treatment (vancomycin, 52%; cotrimoxazole, 27%; rifampin and fusidic acid, 18%). Decolonization was successful in 54 (87%) of the patients in the intent-to-treat analysis and in 51 (98%) of 52 patients in the on-treatment analysis. CONCLUSION: This standardized regimen for MRSA decolonization was highly effective in patients who completed the full decolonization treatment course. PMID- 18510461 TI - Intensive care unit outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae controlled by cohorting patients and reinforcing infection control measures. AB - OBJECTIVE: To describe an outbreak of extended-spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae in the intensive care units (ICUs) of a hospital and the impact of routine and reinforced infection control measures on interrupting nosocomial transmission. DESIGN: Outbreak report. SETTING: A 31-bed intensive care department (composed of 4 ICUs) in a university hospital in Belgium. INTERVENTION: After routine infection control measures (based on biweekly surveillance cultures and contact precautions) failed to interrupt a 2 month outbreak of ESBL-producing K. pneumoniae, reinforced infection control measures were implemented. The frequency of surveillance cultures was increased to daily sampling. Colonized patients were moved to a dedicated 6-bed ICU, where they received cohorted care with the support of additional nurses. Two beds were closed to new admissions in the intensive care department. Meetings between the ICU and infection control teams were held every day. Postdischarge disinfection of rooms was enforced. Broad-spectrum antibiotic use was discouraged. RESULTS: Compared with a baseline rate of 0.44 cases per 1,000 patient-days for nosocomial transmission, the incidence peaked at 11.57 cases per 1,000 patient-days (October and November 2005; rate ratio for peak vs baseline, 25.46). The outbreak involved 30 patients, of whom 9 developed an infection. Bacterial genotyping disclosed that the outbreak was polyclonal, with 1 predominant genotype. Reinforced infection control measures lasted for 50 days. After the implementation of these measures, the incidence fell to 0.08 cases per 1,000 patient-days (rate ratio for after the outbreak vs during the outbreak, 0.11). CONCLUSION: These data indicate that, in an intensive care department in which routine screening and contact precautions failed to prevent and interrupt an outbreak of ESBL-producing K. pneumoniae, reinforced infection control measures controlled the outbreak without major disruption of medical care. PMID- 18510462 TI - Development of a guideline for the management of ventilator-associated pneumonia based on local microbiologic findings and impact of the guideline on antimicrobial use practices. AB - OBJECTIVE: To describe the development of a guideline for the management of ventilator-associated pneumonia (VAP) based on local microbiologic findings and to evaluate the impact of the guideline on antimicrobial use practices. DESIGN: Retrospective comparison of antimicrobial use practices before and after implementation of the guideline. SETTING: Intensive care units at Harborview Medical Center, Seattle, Washington, a university-affiliated urban teaching hospital. PATIENTS: A total of 819 patients who received mechanical ventilation and who underwent quantitative bronchoscopy between July 1, 2003, and June 30, 2005, for suspected VAP. INTERVENTIONS: Implementation of an evidence-based VAP guideline that focused on the use of quantitative bronchoscopy for diagnosis, administration of empirical antimicrobial therapy based on local microbiologic findings and resistance patterns, tailoring definitive antimicrobial therapy on the basis of culture results, and appropriate duration of therapy. RESULTS: During the baseline period, 168 (46.7%) of 360 patients had quantitative cultures that met the diagnostic criteria for VAP, compared with 216 (47.1%) of 459 patients in the period after the guideline was implemented. The pathogens responsible for VAP remained similar between the 2 periods, except that the prevalence of VAP due to carbapenem-resistant Acinetobacter species increased from 1.8% to 15.3% (P<.001), particularly in late-onset VAP. Compared with the baseline period, there was an improvement in antimicrobial use practices after implementation of the guideline: antimicrobial therapy was more frequently tailored on the basis of quantitative culture results (103 [61.3%] of 168 vs 150 [69.4%] of 216 patients; P = .034), there was an increase in the use of appropriate definitive therapy (135 [80.4%] of 168 vs 193 [89.4%] of 216 patients; P = .001), and there was a decrease in the mean duration of therapy (12.0 vs 10.7 days; P = .0014). The all-cause mortality rate was similar in the periods before and after the guideline was implemented (38 [22.6%] of 168 vs 46 [21.3%] of 216 patients; P = .756). CONCLUSIONS: Implementation of a guideline for the management of VAP that incorporated the use of quantitative bronchoscopy, the use of empirical therapy based on local microbiologic findings, tailoring of therapy on the basis of culture results, and use of shortened durations of therapy led to significant improvements in antimicrobial use practices without adversely affecting the all-cause mortality rate. PMID- 18510463 TI - Correlation between respiratory colonization with gram-negative bacteria and development of gram-negative bacterial infection after cardiac surgery. AB - This pilot, observational study involving 286 patients who underwent cardiac surgery found that patients who had endotracheal colonization with gram-negative bacteria at 1 week after surgery were more likely to develop subsequent infection compared to those without colonization (8 of 23 vs. 4 of 40; relative risk 2.3 [95% confidence interval, 1.3-4.1; P value <.05]). PMID- 18510464 TI - Genotypic evolution of Acinetobacter baumannii strains in an outbreak associated with war trauma. AB - The Walter Reed Army Medical Center has experienced an influx of traumatically injured patients either infected or colonized with Acinetobacter baumannii. Using multilocus polymerase chain reaction (PCR) and mass spectrometry to genotype isolates, we found an atypical and evolving strain distribution, distinct from those found at nonmilitary hospitals in the United States. PMID- 18510465 TI - Effect of an infection control program on the frequency of nosocomial viral respiratory infections. AB - We determined the rate of nosocomial viral respiratory infection in infants and the effect of an infection control program during 4 winter seasons. The rate of nosocomial viral respiratory infection decreased from 6.09 episodes per 100 patients admitted during the first study year to 1.46 episodes per 100 patients admitted during the last study year. PMID- 18510466 TI - Using samples to estimate the sensitivity and specificity of a surveillance process. AB - Determining sensitivity and specificity of a postoperative infection surveillance process is a difficult undertaking. Because postoperative infections are rare, vast numbers of negative results exist, and it is often not reasonable to assess them all. This study gives a methodological framework for estimating sensitivity and specificity by taking only a small sample of the number of patients who test negative and comparing their findings to the reference or "gold standard" rather than comparing the findings of all patients to the gold standard. It provides a formula for deriving confidence intervals for these estimates and a guide to minimum requirements for sampling results. PMID- 18510467 TI - Risk factors for infectious spondylodiscitis in patients receiving hemodialysis. AB - A retrospective case-control and cohort analysis of hemodialysis patients was done to identify risk factors for spondylodiscitis. These risk factors included bacteremia, receipt of blood products, invasive procedures, and establishment of vascular access. The death rate was greater for case subjects than for control subjects (odds ratio, 2.7). PMID- 18510468 TI - Nonjudicious dispensing of antibiotics by drug stores in Pratumthani, Thailand. AB - Mock patient presentations of 6 common syndromic ailments to drug stores in Pratumthani, Thailand, were conducted. Appropriate dispensing of antibiotic therapy for all 6 presentations occurred at 56 (20%) of 280 drug stores. By multivariate analysis, drug stores' proximity to a hospital was associated with appropriate dispensing of antibiotics (adjusted odds ratio, 34 [95% confidence interval, 15-83]; P < .001). PMID- 18510469 TI - Effectiveness of alcohol-based hand hygiene gels in reducing nosocomial infection rates. PMID- 18510470 TI - "Cannot detect a change" is not the same as "there is not a change". PMID- 18510471 TI - Trial of alcohol-based hand gel in critical care units. PMID- 18510472 TI - Alcohol-based hand hygiene and nosocomial infection rates. PMID- 18510474 TI - Free radicals in the heart: friend or foe? PMID- 18510475 TI - Nutrition 101: physicians can no longer ignore the healing power of diet and nutritional supplements. PMID- 18510477 TI - Role of tolvaptan in acute decompensated heart failure. AB - Acute decompensated heart failure accounts for more than 1 million hospitalizations in the USA every year. Currently, the most common treatment for symptom relief is the use of loop diuretics, despite recent concerns for potential adverse effects. With the growing understanding of the role of neurohormonal dysregulation in the pathophysiology of heart failure, there has been increasing interest in novel pharmacologic therapies targeting specific neurohormonal axes. Serum arginine vasopressin is a potent vasoconstrictor, as well as an antidiuretic, and serum concentrations are upregulated in heart failure. Tolvaptan, a vasopressin receptor antagonist, has been shown to improve diuresis and symptom relief without adversely affecting renal function, and may be a promising novel therapeutic agent in the growing population of patients with heart failure. PMID- 18510478 TI - Abciximab: a review and update for clinicians. AB - The glycoprotein (GP) IIb/IIIa receptor serves as the final common pathway of platelet-thrombus formation. Thus, the GP IIb/IIIa receptor has been identified as a target for the prevention of thrombus formation during acute coronary syndromes and/or percutaneous coronary intervention. While there are several intravenous GP IIb/IIIa inhibitors available, abciximab has proven to be a dependable agent with unique properties. Based on American College of Cardiology, American Heart Association and Society for Angiography and Interventions guidelines, compared with the other available intravenous GP IIb/IIIa inhibitors, abciximab receives the highest recommendation for use in patients with ST elevation myocardial infarction undergoing primary percutaneous coronary intervention. Abciximab is also unique in that there is no need for renal dosing and its effects are mostly reversible with platelet transfusions. PMID- 18510479 TI - Biomarkers, atherosclerosis and cardiovascular events. AB - The 57th Annual Scientific Session of the American College of Cardiology was held in Chicago (IL, USA) between 29 March and 1 April 2008. It was attended by nearly 30,000 participants from around the world. The conference was highlighted by the presentation of 13 late-breaking clinical trials and 13 late-breaking abstracts. PMID- 18510480 TI - Confusion over thiazolidinedione-induced heart failure: need for a better definition of heart failure. AB - Evaluation of: Erdmann E, Wilcox RG. Weighing up the cardiovascular benefits of thiazolidinedione therapy: the impact of increased risk of heart failure. Eur. Heart J. 29(1), 12-20 (2008). Thiazolidinediones (TZDs) reduce insulin resistance through the modulation of peroxisome proliferator-activated receptor (PPAR)-gamma activity and are, therefore, used for the treatment of individuals with Type 2 diabetes. TZDs have been blamed for inducing heart failure (HF) and are contraindicated in patients with impaired ventricular function. Whether precipitation of HF by TZDs is overestimated or not remains hotly debated in the scientific community. One message from the TZD-HF debacle is that current definitions of HF lack scientific rigour as they fail to assess cardiac organ function directly using a representative and reliable method. Once cardiologists reappraise and update the current definition of HF, appropriate steps can then be taken to answer the question of whether TZDs really induce true HF. PMID- 18510481 TI - Comparison of different LDL apheresis methods. AB - This article presents the generally accepted indications for LDL apheresis treatment. The available LDL apheresis methods differ with respect to acute relative reductions of LDL cholesterol; mean values after the LDL apheresis treatments are not different. Serum triglycerides, HDL-cholesterol, and lipoprotein(a) are also acutely reduced. Available LDL apheresis methods differ with respect to their impact on the coagulation system, on C-reactive protein and on leukocyte count. Cardiovascular events are clearly reduced by the LDL apheresis methods. There is an urgent need to prospectively compare the different LDL apheresis methods taking into account hard end points. The lower target values for LDL cholesterol suggested by international guidelines for high-risk patients will certainly require a more widespread use of LDL apheresis. PMID- 18510482 TI - Gene therapy to prevent occlusion of venous bypass grafts. AB - Revascularization with vein grafts is standard surgical therapy for occlusive arterial diseases. Autologous saphenous vein grafts are important conduits for repairing blocked coronary arteries and are used in the majority of vein graft procedures. Up to 50% of saphenous vein grafts will be occluded during the first decade after surgery. Vein graft occlusion occurs as a result of neointimal hyperplasia, which takes place in response to hemodynamic changes and vessel wall injury, and is characterized by the migration and proliferation of vascular smooth muscle cells. Intimal hyperplasia is further complicated by the concomitant development of atherosclerosis and thrombosis. In the absence of effective pharmacological interventions for the treatment and prevention of occlusive vein graft disease, gene therapy has emerged as a potential therapeutic alternative. Gene therapy could improve vein graft patency by reducing early thrombosis, neointimal hyperplasia and atherosclerosis. In this review we will summarize the emerging applications of gene therapy as a therapeutic tool in occlusive vein graft disease. PMID- 18510483 TI - Biological approaches to ischemic tissue repair: gene- and cell-based strategies. AB - Gene therapy is a potential therapeutic strategy for treatment of ischemic vascular diseases; however, the clinical application of gene therapy has met some anticipated challenges. Recent randomized, controlled trials suggest that patients with cardiovascular disease may also benefit from cell-based therapies, and the optimal treatment regimen may combine both approaches to take advantage of potential synergy between the underlying therapeutic mechanisms. This review discusses recent research into both gene and cell therapy and considers the potential application of a combined treatment approach for cardiovascular and cerebrovascular ischemic diseases. PMID- 18510484 TI - Myocardial repair: from salvage to tissue reconstruction. AB - Cardiac tissue reconstruction following myocardial infarction represents a major challenge in cardiovascular therapy, as current clinical approaches are limited in their ability to regenerate or replace damaged myocardium. Thus, different novel treatments have been introduced aimed at myocardial salvage and repair. Here, we present a review of recent advancements in cardiac cell, gene-based and tissue engineering therapies. Selected strategies in cell therapy and new tools for myocardial gene transfer are summarized. Finally, we consider novel approaches to myocardial tissue engineering as a platform for the integration of various modalities in an attempt to rejuvenate infarcted tissue in vivo. PMID- 18510485 TI - Endothelial progenitor cells: new perspectives and applications in cardiovascular therapies. AB - For over 10 years, bone marrow-derived endothelial progenitor cells (EPCs) have been studied as a novel biomarker to assess the severity of cardiovascular diseases, and as a potential new strategy in regenerative medicine. Cell-based therapy to stimulate postnatal vasculogenesis or to repair vascular integrity is being evaluated for cardiovascular diseases with excess morbidity and mortality, including ischemic heart disease, in-stent restenosis, pulmonary hypertension and peripheral arterial occlusive disease. Although clinical experience is still limited, observed effects appear modest compared with preclinical models. In this review, we will examine major hurdles to the effective use of EPCs, including our incomplete understanding of the characterization and dysfunctional phenotype of circulating EPCs in pathological conditions. Understanding the basic mechanisms of EPC dysfunction will be a prerequisite in enhancing their therapeutic potential. PMID- 18510486 TI - G-CSF- and erythropoietin-based cell therapy: a promising strategy for angiomyogenesis in myocardial infarction. AB - Granulocyte colony-stimulating factor (G-CSF) and erythropoietin are two cytokines that have been demonstrated to improve cardiac function and perfusion in myocardial infarction. G-CSF was initially evaluated as a stem cell mobilizer and erythropoietin as a cytoprotective agent. However, both cytokines have direct cytoprotective effects and stem cell-mobilizing ability. Direct cytoprotective effects of both cytokines are commonly mediated by the Jak-STAT pathway. In preclinical study, G-CSF and erythropoietin improved cardiac function and perfusion by angiomyogenesis and protection of cardiomyocytes in myocardial infarction. However, results from recent clinical trials did not support beneficial effects of cytokine therapy with G-CSF or erythropoietin alone in patients with myocardial infarction. Further studies are required to elucidate the mechanism of action and to improve therapeutic efficacy by employing novel strategies, such as combined cytokines. PMID- 18510487 TI - Surgical management of obstructive hypertrophic cardiomyopathy: the gold standard. AB - While medication is the first line of therapy in obstructive hypertrophic cardiomyopathy, patients who have symptoms refractory to medical treatment or asymptomatic patients with high resting gradients (>or=30 mmHg) may require septal myectomy. Surgical septal myectomy can be performed safely, with excellent survival, relief from symptoms and low morbidity. Alcohol septal ablation is an alternative to surgical treatment, but late outcomes are uncertain. Although both methods of septal reduction relieve left ventricular outflow tract gradients and improve functional status, the need for permanent pacing appears higher with alcohol ablation compared with surgical myectomy. As our understanding of obstructive hypertrophic cardiomyopathy continues to grow, the indications for intervention will evolve. In our practice, septal myectomy remains the gold standard for treatment of obstructive hypertrophic cardiomyopathy. PMID- 18510488 TI - Antioxidant plasma concentration and supplementation in carotid intima media thickness. AB - Cerebrovascular diseases represent a major problem in Western countries. Oxidative stress, an important condition of increased amounts of reactive oxygen species, is now recognized to be a prominent feature of many acute and chronic diseases, and even of the normal aging process. Carotid intima media thickness is an important marker of atherosclerosis that correlates with established coronary heart disease. Changes in carotid intima media thickness, measured by B-mode high resolution carotid ultrasonography, represent an important and early step in carotid plaque formation and progression and are the most common currently used marker to evaluate the progression of atherosclerotic processes. Several therapeutic strategies have been adopted to slow the early atherosclerotic process in asymptomatic subjects in order to reduce the risk of cardiovascular events. An additional step to slow the atherosclerotic process may include interventions to decrease newly emerging coronary risk factors, such as oxidative stress and inflammation. Consuming a diet rich in fruits and vegetables will provide antioxidant vitamins, and carotenoids, which are believed to inhibit tissue damage derived from oxidative processes and may slow the progression of early atherosclerosis, modify the increase in carotid intima media thickness and, consequently, reduce cardiovascular events. This review synthesizes the published literature regarding antioxidant vitamins plasma concentration and supplementation and carotid intima media thickness. PMID- 18510489 TI - Metabolic syndrome and target organ damage: role of blood pressure. AB - A growing body of evidence indicates that the clustering of metabolic and hemodynamic abnormalities characterizing the metabolic syndrome is associated with a prevalence of subclinical damage in a variety of organs, such as left ventricular hypertrophy, thickening or atherosclerotic plaques of carotid arteries, microalbuminuria and deranged renal function. This is clinically relevant since these markers of target organ damage are associated with an increased risk of cardiovascular fatal and nonfatal events. The contribution of the metabolic syndrome to target organ damage in hypertensives is presumably responsible for a substantial increase in cardiovascular fatal and nonfatal events. Thus, target organ damage should be routinely searched for in hypertensives with metabolic syndrome in order to define initial therapeutic strategies and to monitor treatment-induced protection. PMID- 18510490 TI - Cardiorespiratory fitness and sleep-related breathing disorders. AB - Obstructive sleep apnea and central sleep apnea are burgeoning sleep-related breathing disorders within the general population. Most of the associated comorbidities and causes of these sleep disorders are known to negatively impact cardiorespiratory fitness; however, little is known about the direct relationships between cardiorespiratory fitness, obstructive sleep apnea and central sleep apnea. This article provides a systematic analysis of existing peer reviewed, published clinical studies pertaining to the relationship between cardiorespiratory fitness and sleep-related breathing disorders in adults. A brief description of each sleep disorder, the pathophysiology, its epidemiology and its implications for cardiorespiratory fitness are provided. Finally, we discuss therapy for each disorder and its effect on the cardiovascular system. PMID- 18510491 TI - Current possibilities of ACE inhibitor and ARB combination in arterial hypertension and its complications. AB - The renin-angiotensin-aldosterone system (RAAS) plays a crucial role in blood pressure regulation and hypertension-related complications. Angiotensin converting enzyme inhibitors (ACEIs) were the first to be used to block the RAAS and now have many compelling indications in the treatment of hypertension and its cardiovascular and renal complications. Angiotensin II receptor blockers (ARBs), introduced 20 years later, have been shown to be equally as effective as antihypertensive treatment and are also associated with a lower number of side effects. Furthermore, in clinical trials ARBs and ACEIs were associated with comparable benefits for their most typical indications. This was confirmed in the 2007 New European Society of Hypertension/European Society of Cardiology (ESH/ESC) guidelines for the management of hypertension by comparable specific recommendations for ARB and ACEI treatment. There is sufficient theoretical background and, in some cases, also clinical evidence that combination therapy with ACEIs and ARBs may be more beneficial than monotherapy with either of the groups alone, both in uncomplicated hypertension and with concomitant heart failure or renal dysfunction. However, the combination of ACEI and ARB was not recommended in the ESH/ESC 2007 Guidelines. This may change after the publication of the Ongoing Telmisartan Alone and in Combination with Ramipril Global End point Trial (ONTARGET) study, the preliminary results of which have just been presented. In heart failure, recent studies have shown that the combination of ACEI and ARB decreases cardiovascular mortality and the number of hospitalizations due to aggravation of heart failure. These results have been reflected in the newest ESC guidelines of the heart failure treatment. Nephroprotective properties of the combination of ACEs and ARBs have been proved both in studies on nondiabetic and diabetic nephropathy. The potential benefits, indications in prespecified groups of patients, the most recent data from clinical trials and latest research regarding dual blockade of RAAS will be reviewed in this article. PMID- 18510494 TI - WITHDRAWN:Involvement of JAK/STAT, PI3K, PKC and MAP kinases in the growth hormone induced production of cytokines in murine peritoneal macrophages in vitro. AB - The Ahead of Print article entitled "Involvement of JAK/STAT, PI3K, PKC and MAP kinases in the growth hormone induced production of cytokines in murine peritoneal macrophages in vitro", which was published online on 30 May 2008, was withdrawn at the author's request on 8 October 2008. PMID- 18510493 TI - Modulation of hepatocyte nuclear factor-4alpha function by the peroxisome proliferator-activated receptor-gamma co-activator-1alpha in the acute-phase response. AB - HNF-4alpha (hepatocyte nuclear factor-4alpha) is a key regulator of liver specific gene expression. To understand the mechanisms governing the regulation of HNF-4alpha function during the APR (acute-phase response), the effects of transcription co-activators, including p300, PGC-1alpha (peroxisome-proliferator activated receptor-gamma co-activator-1alpha) and SRC (steroid receptor co activator)-1alpha were investigated in an injury cell model. We have shown previously that the HNF-4alpha-sensitive APR genes ApoB (apolipoprotein B), TTR (transthyretin) and alpha1-AT (alpha1-antitrypsin) were regulated at the DNA binding and transcriptional levels after cytokine stimulation. We now show that co-activators have a differential impact on the transactivation of HNF-4alpha sensitive genes via HNF-4alpha-binding sites in ApoB, TTR or alpha1-AT promoters. PGC-1alpha strongly enhances the transactivation of ApoB and alpha1-AT and, to a lesser extent, of TTR, whereas SRC-1alpha and p300 only have a weak or no effect on these three genes. More importantly, it was found that PGC-1alpha has a novel role in the modulation of the binding ability of HNF-4alpha in response to cytokine treatment. Using in vitro and in vivo approaches, electrophoretic mobility-shift and chromatin immunoprecipitation assays, we demonstrate that the reduced HNF-4alpha-DNA binding ability induced by cytokines is eliminated by overexpression of PGC-1alpha. Cytokine treatment does not significantly alter the protein levels of HNF-4alpha and PGC-1alpha, but it does reduce the recruitment of PGC-1alpha to HNF-4alpha-binding sites and thereby decreases transcriptional activity. These results establish the importance of PGC-1alpha for HNF-4alpha function and describe a new HNF-4alpha-dependent regulatory mechanism that is involved in the response to injury. PMID- 18510495 TI - Statistical significance, power and sample size - what does it all mean? PMID- 18510496 TI - The role of nutrients in modulating disease. AB - The role of nutrition in the management of diseases has often centred on correcting apparent nutrient deficiencies or meeting estimated nutritional requirements of patients. Nutrition has traditionally been considered a supportive measure akin to fluid therapy and rarely it has been considered a primary means of ameliorating diseases. Recently, however, further understanding of the underlying mechanisms of various disease processes and how certain nutrients possess pharmacological properties have fuelled an interest in exploring how nutritional therapies themselves could modify the behaviour of various conditions. Nutrients such as omega-3 fatty acids, antioxidants and certain amino acids such as arginine and glutamine have all been demonstrated to have at least the potential to modulate diseases. Developments in the area of critical care nutrition have been particularly exciting as nutritional therapies utilising a combination of approaches have been shown to positively impact outcome beyond simply proving substrate for synthesis and energy. Application of certain nutrients for the modulation of diseases in veterinary patients is still in early stages, but apparent successes have already been demonstrated, and future studies are warranted to establish optimal approaches. This review describes the rationale of many of these approaches and discusses findings both in human beings and in animals, which may guide future therapy. PMID- 18510498 TI - Methimazole-triggered lymphadenomegaly in a hyperthyroid cat? PMID- 18510501 TI - Synthetic morphology: prospects for engineered, self-constructing anatomies. AB - This paper outlines prospects for applying the emerging techniques of synthetic biology to the field of anatomy, with the aim of programming cells to organize themselves into specific, novel arrangements, structures and tissues. There are two main reasons why developing this hybrid discipline--synthetic morphology- would be useful. The first is that having a way to engineer self-constructing assemblies of cells would provide a powerful means of tissue engineering for clinical use in surgery and regenerative medicine. The second is that construction of simple novel systems according to theories of morphogenesis gained from study of real embryos will provide a means of testing those theories rigorously, something that is very difficult to do by manipulation of complex embryos. This paper sets out the engineering requirements for synthetic morphology, which include the development of a library of sensor modules, regulatory modules and effector modules that can be connected functionally within cells. A substantial number of sensor and regulatory modules already exist and this paper argues that some potential effector modules have already been identified. The necessary library may therefore be within reach. The paper ends by suggesting a set of challenges, ranging from simple to complex, the achievement of which would provide valuable proofs of concept. PMID- 18510502 TI - Spatial packing, cranial base angulation, and craniofacial shape variation in the mammalian skull: testing a new model using mice. AB - The hypothesis that variation in craniofacial shape within and among species is influenced by spatial packing has a long history in comparative anatomy, particularly in terms of primates. This study develops and tests three alternative models of spatial packing to address how and to what extent the cranial base angle is influenced by variation in brain and facial size. The models are tested using mouse strains with different mutations affecting craniofacial growth. Although mice have distinctive crania with small brains, long faces, and retroflexed cranial bases, the results of the study indicate that the mouse cranial base flexes to accommodate larger brain size relative to cranial base length. In addition, the mouse cranial base also extends, but to a lesser degree, to accommodate larger face size relative to cranial base length. In addition, interactions between brain size, face size, and the widths and lengths of the components of the cranial base account for a large percentage of variation in cranial base angle. The results illustrate the degree to which the cranial base is centrally embedded within the covariation structure of the craniofacial complex as a whole. PMID- 18510503 TI - Cranial performance in the Komodo dragon (Varanus komodoensis) as revealed by high-resolution 3-D finite element analysis. AB - The Komodo dragon (Varanus komodoensis) displays a unique hold and pull-feeding technique. Its delicate 'space-frame' skull morphology differs greatly from that apparent in most living large prey specialists and is suggestive of a high degree of optimization, wherein use of materials is minimized. Here, using high resolution finite element modelling based on dissection and in vivo bite and pull data, we present results detailing the mechanical performance of the giant lizard's skull. Unlike most modern predators, V. komodoensis applies minimal input from the jaw muscles when butchering prey. Instead it uses series of actions controlled by postcranial muscles. A particularly interesting feature of the performance of the skull is that it reveals considerably lower overall stress when these additional extrinsic forces are added to those of the jaw adductors. This remarkable reduction in stress in response to additional force is facilitated by both internal and external bone anatomy. Functional correlations obtained from these analyses also provide a solid basis for the interpretation of feeding ecology in extinct species, including dinosaurs and sabre-tooth cats, with which V. komodoensis shares various cranial and dental characteristics. PMID- 18510504 TI - Osteology and taxonomic revision of Hyphalosaurus (Diapsida: Choristodera) from the Lower Cretaceous of Liaoning, China. AB - Although the long-necked choristodere Hyphalosaurus is the most abundant tetrapod fossil in the renowned Yixian Formation fossil beds of Liaoning Province, China, the genus has only been briefly described from largely unprepared specimens. This paper provides a thorough osteological description of the type species Hyphalosaurus lingyuanensis and the con-generic species Hyphalosaurus baitaigouensis based on the study of fossils from several research institutions in China. The diagnoses for these two species are revised based on comparison of a large sample of specimens from the type area and horizon of each of the two species. The skull, better known in H. baitaigouensis, exhibits key choristodere synapomorphies including an elongate contact between the prefrontals and posteriorly expanded supratemporal fenestrae that strongly support the placement of the highly derived hyphalosaurids within Choristodera. Both species of Hyphalosaurus share a proportionally small head, an elongate neck, a relatively unspecialized appendicular skeleton and a long, dorsoventrally heightened tail. Soft tissue preservation in several specimens provides rare insight into the integument of an extinct group. The integument of Hyphalosaurus is made up of small polygonal scales with several parasagittal rows of large, keeled, ovoid scutes. These possibly ornamental scutes bear a strong resemblance to the rows of large scutes in the monjurosuchid choristodere Monjurosuchus splendens. Observations from a variety of growth stages reveal that significant ontogenetic change in the proportions of the body and limb bones occurred in both species of Hyphalosaurus. PMID- 18510505 TI - Calibration of estimated biting forces in domestic canids: comparison of post mortem and in vivo measurements. AB - Estimates of biting forces are widely used in paleontological and comparative studies of feeding mechanics and performance, and are usually derived from lever models based on measurements made on the skull that are relevant to the mechanics of the masticatory system. Owing to assumptions and unmeasurable errors in their estimation, such values are used comparatively rather than as absolute estimates. The purpose of this paper was to provide calibration of post-mortem calculated bite force estimates by comparing them to in vivo forces derived from a sample of 20 domestic dogs (Canis familiaris) during muscle stimulation under general anaesthesia. Two lever models previously described in the literature were used to estimate post-mortem values, and regression analysis was also performed to derive best-fit equations against a number of morphometric measurements on the skull. The ranges of observed forces in vivo were 147-946 N at the canine, and 524-3417 N at the second molar. The lever models substantially underestimated these forces, giving mean values between 39% and 61% of the observed means. Predictability was considerably improved by removing the linear bias and deviation of the regression slope from unity with an adjustment equation. Best fit statistical models developed on these animals performed considerably better (calculated means within 0.54% of observed means) and included easily measureable variables such as bodyweight, dimensions of the temporalis fossa and out-lever from the jaw joint to the biting tooth. These data should lead to more accurate absolute, rather than relative, estimates of biting forces for other extant and fossil canids, and other carnivorans by extrapolation. PMID- 18510506 TI - The anatomical relationships between the avian eye, orbit and sclerotic ring: implications for inferring activity patterns in extinct birds. AB - Activity pattern, or the time of day when an animal is awake and active, is highly associated with that animal's ecology. There are two principal activity patterns: diurnal, or awake during the day in a photopic, or high light level, environment; and nocturnal, awake at night in scotopic, or low light level, conditions. Nocturnal and diurnal birds exhibit characteristic eye shapes associated with their activity pattern, with nocturnal bird eyes optimized for visual sensitivity with large corneal diameters relative to their eye axial lengths, and diurnal birds optimized for visual acuity, with larger axial lengths of the eye relative to their corneal diameters. The current study had three aims: (1) to quantify the nature of the relationship between the avian eye and its associated bony anatomy, the orbit and the sclerotic ring; (2) to investigate how activity pattern is reflected in that bony anatomy; and (3) to identify how much bony anatomy is required to interpret activity pattern reliably for a bird that does not have the soft tissue available for study, specifically, for a fossil. Knowledge of extinct avian activity patterns would be useful in making palaeoecological interpretations. Here eye, orbit and sclerotic ring morphologies of 140 nocturnal and diurnal bird species are analysed in a phylogenetic context. Although there is a close relationship between the avian eye and orbit, activity pattern can only be reliably interpreted for bony-only specimens, such as a fossil, that include both measurements of the sclerotic ring and orbit depth. Any missing data render the fossil analysis inaccurate, including fossil specimens that are flat and therefore do not have an orbit depth available. For example, activity pattern cannot be determined with confidence for Archaeopteryx lithographica, which has a complete sclerotic ring but no orbit depth measurement. Many of the bird fossils currently available that retain a good sclerotic ring tend to be flat specimens, while three-dimensionally preserved bird fossils tend not to have a well-preserved sclerotic ring or a well-defined optic foramen, necessary for delimiting the orbit depth. PMID- 18510508 TI - Immunohistochemical distribution of regulatory peptides in the human fetal adenohypophysis. AB - We have studied here the cellular distribution of several regulatory peptides in hormone-producing cells of the human pituitary during the fetal period. Immunohistochemistry was used to show the expression of several regulatory peptides, namely Angiotensin-II, Neurotensin and Galanin, at successive gestational stages and their co-localization with hormones in the human fetal adenohypophysis. Somatotrophs, gonadotrophs and thyrotrophs were differentiated earliest. At gestational week 9, Angiotensin-II immunoreactivity was co-localized only with growth hormone immunoreactivity in somatotrophs, one of the first hormone-producing cells to differentiate. This co-localization remained until week 37. Neurotensin immunoreactivity was present in gonadotrophs and thyrotrophs in week 23, after FSH and TSH hormone differentiation. Galanin immunoreactivity was present in all hormone-producing cell types except corticotrophs. The different pro-opiomelanocortin-derived peptides were detected at different stages of gestation and adrenocorticotrophic hormone immunoreaction was the last to be detected. Our results show an interesting relationship between regulatory peptides and hormones during human fetal development, which could imply that these peptides play a regulatory role in the development of pituitary function. PMID- 18510507 TI - The presence and absence of prosencephalic cell groups relaying striatal information to the medial and lateral thalamus in tenrec. AB - Although there are remarkable differences regarding the output organization of basal ganglia between mammals and non-mammals, mammalian species with poorly differentiated brain have scarcely been investigated in this respect. The aim of the present study was to identify the pallidal neurons giving rise to thalamic projections in the Madagascar lesser hedgehog tenrec (Afrotheria). Following tracer injections into the thalamus, retrogradely labelled neurons were found in the depth of the olfactory tubercle (particularly the hilus of the Callejal islands and the insula magna), in subdivisions of the diagonal band complex, the peripeduncular region and the thalamic reticular nucleus. No labelled cells were seen in the globus pallidus. Pallidal neurons were tentatively identified on the basis of their striatal afferents revealed hodologically using anterograde axonal tracer substances and immunohistochemically with antibodies against enkephalin and substance P. The data showed that the tenrec's medial thalamus received prominent projections from ventral pallidal cells as well as from a few neurons within and ventral to the cerebral peduncle. The only regions projecting to the lateral thalamus appeared to be the thalamic reticular nucleus (RTh) and the dorsal peripeduncular nucleus (PpD). On the basis of immunohistochemical data and the topography of its thalamic projections, the PpD was considered to be an equivalent to the pregeniculate nucleus in other mammals. There was no evidence of entopeduncular (internal pallidal) neurons being present within the RTh/PpD complex, neuropils of which did not stain for enkephalin and substance P. The ventrolateral portion of RTh, the only region eventually receiving a striatal input, projected to the caudolateral rather than the rostrolateral thalamus. Thus, the striatopallidal output organization in the tenrec appeared similar, in many respects, to the output organization in non-mammals. This paper considers the failure to identify entopeduncular neurons projecting to the rostrolateral thalamus in a mammal with a little differentiated cerebral cortex, and also stresses the discrepancy between this absence and the presence of a distinct external pallidal segment (globus pallidus). PMID- 18510509 TI - Rapid loss of motor nerve terminals following hypoxia-reperfusion injury occurs via mechanisms distinct from classic Wallerian degeneration. AB - Motor nerve terminals are known to be vulnerable to a wide range of pathological stimuli. To further characterize this vulnerability, we have developed a novel model system to examine the response of mouse motor nerve terminals in ex vivo nerve/muscle preparations to 2 h hypoxia followed by 2 h reperfusion. This insult induced a rapid loss of neurofilament and synaptic vesicle protein immunoreactivity at pre-synaptic motor nerve terminals but did not appear to affect post-synaptic endplates or muscle fibres. The severity of nerve terminal loss was dependent on the age of the mouse and muscle type: in 8-12-week-old mice the predominantly fast-twitch lumbrical muscles showed an 82.5% loss, whereas the predominantly slow-twitch muscles transversus abdominis and triangularis sterni showed a 57.8% and 27.2% loss, respectively. This was contrasted with a > 97% loss in the predominantly slow-twitch muscles from 5-6-week-old mice. We have also demonstrated that nerve terminal loss occurs by a mechanism distinct from Wallerian degeneration, as the slow Wallerian degeneration (Wld(s)) gene did not modify the extent of nerve terminal pathology. Together, these data show that our new model of hypoxia-reperfusion injury is robust and repeatable, that it induces rapid, quantitative changes in motor nerve terminals and that it can be used to further examine the mechanisms regulating nerve terminal vulnerability in response to hypoxia-reperfusion injury. PMID- 18510510 TI - Morphofunctional responses to anaemia in rat skeletal muscle. AB - Adult male Sprague-Dawley rats were randomly assigned to two groups: control and anaemic. Anaemia was induced by periodical blood withdrawal. Extensor digitorum longus and soleus muscles were excised under pentobarbital sodium total anaesthesia and processed for transmission electron microscopy, histochemical and biochemical analyses. Mitochondrial volume was determined by transmission electron microscopy in three different regions of each muscle fibre: pericapillary, sarcolemmal and sarcoplasmatic. Muscle samples sections were also stained with histochemical methods (SDH and m-ATPase) to reveal the oxidative capacity and shortening velocity of each muscle fibre. Determinations of fibre and capillary densities and fibre type composition were made from micrographs of different fixed fields selected in the equatorial region of each rat muscle. Determination of metabolites (ATP, inorganic phosphate, creatine, creatine phosphate and lactate) was done using established enzymatic methods and spectrophotometric detection. Significant differences in mitochondrial volumes were found between pericapillary, sarcolemmal and sarcoplasmic regions when data from animal groups were tested independently. Moreover, it was verified that anaemic rats had significantly lower values than control animals in all the sampled regions of both muscles. These changes were associated with a significantly higher proportion of fast fibres in anaemic rat soleus muscles (slow oxidative group = 63.8%; fast glycolytic group = 8.2%; fast oxidative glycolytic group = 27.4%) than in the controls (slow oxidative group = 79.0%; fast glycolytic group = 3.9%; fast oxidative glycolytic group = 17.1%). No significant changes were detected in the extensor digitorum longus muscle. A significant increase was found in metabolite concentration in both the extensor digitorum longus and soleus muscles of the anaemic animals as compared to the control group. In conclusion, hypoxaemic hypoxia causes a reduction in mitochondrial volumes of pericapillary, sarcolemmal, and sarcoplasmic regions. However, a common proportional pattern of the zonal distribution of mitochondria was maintained within the fibres. A significant increment was found in the concentration of some metabolites and in the proportion of fast fibres in the more oxidative soleus muscle in contrast to the predominantly anaerobic extensor digitorum longus. PMID- 18510511 TI - The effects of high-fat diet on the renal structure and morphometric parametric of kidneys in rats. AB - To characterize the kidney in a high-fat-induced obesity model, we examined the renal structure of adult Sprague-Dawley rats fed a control diet or a high-fat diet for 3 months. Ten adult female Sprague-Dawley rats were fed a diet consisting highly of fat (30%) for a period of 3 months. Ten control rats were maintained with standard rat chow. All animals were weighed every 10 days for 3 months. At the end of the experiment, the naso-anal length of the anaesthetized rats was measured to calculate body mass index, and subsequently whole kidneys of intracardially formalin-perfused animals were removed. Quantitative features of the kidney were analysed with the Cavalieri and physical dissector methods applied to serial paraffin sections. Kidney samples were also examined histologically. The body mass indices of the control and treatment groups were 4.528 +/- 0.242 and 5.876 +/- 0.318 kg m(-2), respectively. The difference between the body mass indices of the two groups was statistically significant (P < 0.01, Mann-Whitney U-test), suggesting that the animals fed with a high-fat diet may be overweight. Stereological examination of the kidneys revealed differences in kidney weight, total kidney volume, volume of cortex, medulla, glomeruli, proximal and distal tubules, and numerical density of glomeruli and glomerular height in the treatment group compared with the control group. Light microscopic investigation showed a dilatation in blood vessels and Bowman's space, mononuclear cell infiltration, degeneration in nephrons, including glomerulosclerosis and tubular defects, and an increase in the connective tissue in the kidneys in the treatment group. We concluded that a fatty diet is responsible for the rats' obesity and may lead to renal deformities as a result of histopathological changes such as dilatation, tubular defects, inflammation and connective tissue enlargement of the kidney. PMID- 18510513 TI - Image processing techniques to quantify microprojections on outer corneal epithelial cells. AB - It is widely accepted that cellular microprojections (microvilli and/or microplicae) of the corneal surface are essential to maintain the functionality of the tissue. To date, the characterization of these vital structures has been made by analysing scanning or transmission electron microscopy images of the cornea by methods that are intrinsically subjective and imprecise (qualitative or semiquantitative methods). In the present study, numerical data concerning three microprojection features were obtained by an automated method and analysed to establish which of them showed less variability. We propose that the most stable microprojection characteristic would be a useful sign in early detection of epithelial damage or disease. With this aim, the scanning electron microscopy images of 220 corneal epithelial cells of nine rabbits were subjected to several image processing techniques to quantify microprojection density, microprojection average size and surface covered by microprojections (SCM). We then assessed the reliability of the methods used and performed a statistical analysis of the data. Our results show that the thresholding process, the basis of all image processing techniques used in this work, is highly reliable in separating microprojections from the rest of the cell membrane. Assessment of histogram information from thresholded images is a good method to quantify SCM. Amongst the three studied variables, SCM was the most stable (with a coefficient of variation of 15.24%), as 89.09% of the sample cells had SCM values > or = 40%. We also found that the variability of SCM was mainly due to intercellular differences (the cell factor contribution represented 88.78% of the total variation in the analysed cell areas). Further studies are required to elucidate how healthy corneas maintain high SCM values. PMID- 18510512 TI - Quantification of diverse subcellular immunohistochemical markers with clinicobiological relevancies: validation of a new computer-assisted image analysis procedure. AB - Tissue microarray technology and immunohistochemical techniques have become a routine and indispensable tool for current anatomical pathology diagnosis. However, manual quantification by eye is relatively slow and subjective, and the use of digital image analysis software to extract information of immunostained specimens is an area of ongoing research, especially when the immunohistochemical signals have different localization in the cells (nuclear, membrane, cytoplasm). To minimize critical aspects of manual quantitative data acquisition, we generated semi-automated image-processing steps for the quantification of individual stained cells with immunohistochemical staining of different subcellular location. The precision of these macros was evaluated in 196 digital colour images of different Hodgkin lymphoma biopsies stained for different nuclear (Ki67, p53), cytoplasmic (TIA-1, CD68) and membrane markers (CD4, CD8, CD56, HLA-Dr). Semi-automated counts were compared to those obtained manually by three separate observers. Paired t-tests demonstrated significant differences between intra- and inter-observer measurements, with more substantial variability when the cellular density of the digital images was > 100 positive cells/image. Overall, variability was more pronounced for intra-observer than for inter observer comparisons, especially for cytoplasmic and membrane staining patterns (P < 0.0001 and P = 0.050). The comparison between the semi-automated and manual microscopic measurement methods indicates significantly lower variability in the results yielded by the former method. Our semi-automated computerized method eliminates the major causes of observer variability and may be considered a valid alternative to manual microscopic quantification for diagnostic, prognostic and therapeutic purposes. PMID- 18510515 TI - WFH--the cornerstone of global development: 45 years of progress. AB - The World Federation of Hemophilia (WFH) has been the cornerstone of global development for 45 years. The WFH has identified and optimized the essential elements of a model for the development of a sustainable national care programme. The five elements of the WFH Development Model are integrated and interdependent: ensuring accurate laboratory diagnosis, achieving government support for a national programme, improving the care delivery system, increasing the availability of treatment products and building a strong national patient organization. It can been demonstrated that patient organizations, healthcare providers and the Ministry of Health working together in coalition is essential to achieving sustainable care. Equally important, the provision of care by a multidisciplinary team of trained professionals within a comprehensive care setting is fundamentally important to optimize outcomes. Using data from the WFH Global Survey, it is evident that the WFH Development Model brings about sustainable improvements in care. To support the Model, the WFH has created a vast range of tools, guides and programmes tailored to specific development needs. The Global Alliance for Progress is the preeminent WFH development program. Five years of outcomes data document a narrowing of the care gap between developed and developing nations. To ensure the continued advance towards the WFH vision of Treatment for All, it is vital that global collaboration occur on the research front as well. The WFH is well positioned to meet the challenges ahead and to continue serving as the cornerstone of global collaboration and development. PMID- 18510516 TI - Back to the future: a recent history of haemophilia treatment. AB - In the last few decades, the management of patients with haemophilia has witnessed dramatic improvements, through the larger availability of safe plasma derived and recombinant products for replacement therapy. Another important step forward is the progressively larger-scale implementation of primary prophylaxis in children. Currently, the main problem in patients with haemophilia is the onset of antibodies inactivating the infused clotting factor (inhibitors), even though immune tolerance regimens that are able to eradicate inhibitors and the availability of products that bypass the intrinsic coagulation defects have dramatically improved the management of these patients. Cure of haemophilia through gene transfer is being attempted, but relatively, it is far from being implemented on a large scale. It is likely that further improvements in replacement therapy will occur in the near future, through the availability of new-therapeutic tools such as factors VIII and IX with longer half-lives, more potent bypassing agents and factors extracted from the milk of transgenic animals. PMID- 18510517 TI - Quality of life in haemophilia. AB - There is a growing interest in patient-reported outcomes as measures for evaluating the benefits of new and existing treatments. Health-related quality of life (HRQoL) is one of these patient-reported outcomes and represents the individual experience and perception of illness/health together with the psychosocial response to disease-related and treatment-related symptoms. Generic and disease-specific HRQoL questionnaires enable us to assess and quantify the multi-dimensional perception of well-being, namely the physical components and the psychological (emotional, mental, social and behavioural) components of patient's well-being and functioning. These instruments should be standardized and validated and they should prove to be reliable, valid, specific and sensitive in a similar manner to instruments created for objective parameters. HRQoL assessment can help us to evaluate the benefits of new treatments from the perspective of patient's values and expectations. It can also help to evaluate the quality of care provided, in order to be able to improve it at a local and national level. Moreover, HRQoL assessment can be routinely assessed to monitor improvement and progress or deterioration and decline from the global point of view of each single patient, integrating the otherwise limited angle of objective signs and instrumental or lab parameters. PMID- 18510518 TI - Management of hepatitis virus infections. PMID- 18510519 TI - How to get rid of inhibitors. PMID- 18510520 TI - Inhibitor development. AB - The immune response to factor VIII and the development of inhibitory antibodies is a complex multi-factorial process involving a variety of immune regulatory genes and cells, several of which have the potential to determine risk. A better understanding of the mechanisms involved will increase the likelihood of development of new therapeutic options for patients with hemophilia. This review summarizes genetic and non-genetic risk factors currently under evaluation, and the potential modulative effect of the von Willebrand factor on factor VIII immuno- and antigenicity. In addition, the role of T-regulatory cells in the pathogenicity of inhibitors will be discussed. PMID- 18510521 TI - Linking the world with training and research for improving haemophilia care. PMID- 18510522 TI - Epidemiology and general guidelines of the management of acquired haemophilia and von Willebrand syndrome. PMID- 18510523 TI - von Willebrand disease update: diagnostic and treatment dilemmas. AB - Although von Willebrand disease (VWD) is now well-described, many facets of diagnosis and management continue to be debated. The diagnosis of type 1 disease can be difficult but recent genetic analyses help to distinguish many factors which can influence von Willebrand factor (VWF) levels and bleeding phenotype. Type 2 disease (functional abnormalities) includes a particularly interesting group of disorders with faulty binding between VWF and FVIIIC (Normandy) where treatment methods need careful consideration. Type 3 VWD is the most severe form of VWD and a new international study is underway to examine the use of prophylaxis. PMID- 18510524 TI - Animal models of bleeding and tissue repair. AB - While a number of animal models have been developed for human haemophilia, it has been difficult to develop reproducible measures of bleeding in these models. They have also not been extensively utilized to study the complications of haemophilia beyond blood loss. Poor haemostatic function also leads to local haematomas, joint damage and poor wound healing. Some of the abnormalities related to bleeding are because of the deleterious effects of iron deposition in the tissues. Evidence from mouse skin wound and joint haemorrhage models suggests that bleeding and iron deposition initiate a vicious cycle of inflammation, angiogenesis and renewed bleeding. However, there is much yet to be learned about the effects of bleeding on tissue responses, including validating the results of animal studies in clinical trials. PMID- 18510525 TI - Relevance of quantitative assessment of bleeding in haemorrhagic disorders. AB - In the last few years, there has been a growing interest in the diagnosis of mild bleeding disorders (MBD) to find reliable tools for the assessment of their inherent bleeding risk and minimum criteria for the definition of a clinically useful diagnosis. Unlike in more severe haemorrhagic disorders, in MBD, the bleeding history may overlap with that reported by normal people. This problem has required the development of strategies that could allow the assessment of bleeding symptoms from both a qualitative (presence or absence) and quantitative (bleeding severity) aspect. An example of high quality clinical research in bleeding disorders was given by the systematic approach used for the evaluation of menorrhagia. For this symptom, the most common in women with bleeding disorders, the use of pictorial charts provided many new insights. Dr Kadir will review its use in a clinical context. The assessment of the whole bleeding history requires first, the development of reproducible tools to collect symptoms and secondly, formulation of easily applicable criteria to convert the collected data into clinical information. Dr Tosetto will propose a bleeding questionnaire in which clinical criteria were developed and validated, and show how a summative, quantitative index of bleeding severity (the Bleeding Score) could be used in von Willebrand disease. Finally, Dr James will review the development of quantitative analysis in children, a particularly important and difficult application, but one that needs to be tackled urgently. PMID- 18510526 TI - Diagnosis of factor VIII deficiency. AB - The correct diagnosis of factor VIII deficiency and the assessment of severity of the disease are essential for a patient-tailored treatment strategy. An optimal diagnostic procedure comprises sensitive and specific screening methods and factor VIII activity assays. Different screening reagents show variable characteristics and receiver operator characteristic curves are presented showing the relation between sensitivity and specificity of eleven activated partial thromboplastin time reagents. The details of the three methods for factor VIII activity assay, one-stage and two-stage assay and chromogenic assays, are discussed. The chromogenic assay seems to be more sensitive than the one-stage assay with regard to the detection of severe haemophilia. Discrepant results obtained with one-stage and two-stage assays are reviewed and discussed. PMID- 18510527 TI - New assays for monitoring haemophilia treatment. AB - Precise measurements of factor VIII (FVIII) or factor IX (FIX) activity are believed to be essential for clinical management in haemophilia, although discrepancies between factor levels and clinical severity have been recognized. Clot wave form analysis has demonstrated that different wave form patterns may be evident in severe haemophilia A patients with levels of FVIII activity <1 IU dL( 1), and this might explain, in part, the phenotypic heterogeneity seen in these patients. In addition, the relatively new technique of computer-assisted thrombelastography (TEG), in which coagulation is initiated by tissue factor, has revealed a considerable degree of variability in different patients in the presence FVIII levels, which are sufficient to normalize TEG parameters. In contrast, a global thrombin generation test (TGT) has been proposed as a sensitive and reliable method for assessing overall clotting function in haemophilia patients. Several studies have demonstrated a significant correlation between TGT and FVIII/FIX levels, and these measurements also appear to correlate with the clinical phenotype. The TGT may be very useful, therefore, for evaluating overall haemostasis in different clinical situations, although substantial inter-assay and inter-individual variations have been reported. Both the TEG and TGT have been found to be particularly helpful for monitoring haemostatic therapy with bypassing agents or conventional FVIII or FIX concentrates in patients with inhibitors. These global tests enable the selection of appropriate therapeutic agents in individual circumstances and offer the opportunity to tailor the most effective haemostatic treatment even during severe bleeding or major surgery. PMID- 18510528 TI - Laboratory issues in bleeding disorders. AB - Selected laboratory issues critical for the appropriate diagnosis of haemophilia A and B, von Willebrand's disease (VWD) and more rare bleeding disorders (RBD) are discussed from a worldwide perspective. The overall picture that emerges is on the whole reassuring. Even in non-Western countries like Latin America, most cases of haemophilia are appropriately diagnosed. Moreover, national and international laboratory training workshops are further improving the diagnostic capabilities also in less severe disorders. Most of the RBD can be appropriately diagnosed with relatively simple tests wherever a high clinical suspicion is present. Moreover, minimal requirements for a useful clinical diagnosis are not too far from the capabilities of majority of non-Western countries. The most needed areas concern VWD and platelet function disorders, which suffer from inadequate diagnostic standardization, hampering widespread diagnostic capability in both Western and non-Western countries. PMID- 18510529 TI - New approaches in the measurement of coagulation. AB - In this session contributors present recent developments in laboratory tools for investigation of haemorrhagic disorders as well as their relative utility in clinical research. In an overview of B. Sorensen the present knowledge is summarized on the dynamic properties of whole blood fibrin formation as studied by changes in whole blood elasticity on a thrombelastometry system. Additionally, fibrin formation dynamics using simple APTT methods are presented. G. Castaman reviews the pathophysiology of von Willebrand's disease (VWD) and explains which tests are best used in diagnosis and subclassification of VWD accounting for recent developments. This presentation also describes the treatment technologies available today and their implications in clinical management of bleeding episodes in VWD. J. Lloyd addresses the assay discrepancy phenomenon that is found in some of our patients suffering from mild haemophilia A. Assay discrepancy most often means a much lower factor VIII:C value by a two-stage or chromogenic assay for factor VIII:C compared to the activity recorded by the one stage clotting system for factor VIII:C. In rare cases, the opposite phenomenon exist. The presentation includes data from 16 Australian families with discrepant results. D. Varon reports on an assay for study of platelet function in whole blood under flow conditions. The equipment is described as a cone-and-plate(let) analyser in which the adhesion and aggregation of platelets onto a polystyrene surface is studied under arterial flow conditions. Basically, it is anticipated that proteins such as VWF and fibrinogen of flowing blood is attached to the polystyrene surface where they build up a thrombogenic surface. In the study of the author platelets were pre-activated with agonists and platelet deposits were determined after passage of whole blood for a pre-set time interval. Data presented suggest that the assay is sensitive to platelet numbers as well as qualitative changes in platelets themselves, and several examples of disorders characterized by enhanced as well as reduced platelet aggregating activities illustrates the sensitivity of the method. PMID- 18510530 TI - Genetic aspects and research development in haemostasis. PMID- 18510531 TI - Advances in minor oral surgery in patients with congenital bleeding disorders. PMID- 18510532 TI - Ethical considerations in clinical investigation: exploring relevance in haemophilia research. AB - Painful controversy has so far been largely absent from the history of haemophilia-related clinical research. However, the investigative methods now needed to realize evidence-based clinical practice, therapeutic advance, and a progressive standard of care for patients worldwide will be accompanied by the potential for ethical dilemma and transgression. From the current vantage point, three primary ethical issues merit special consideration: (i) the therapeutic misconception inherent to all clinical research and the randomized trial in particular; (ii) high risk and potentially non-beneficial novel technology research in children; and (iii) a collaborative partnership approach to research in the developing world. This study will focus on a discussion of each of these, drawing from the research ethics literature to offer a potential template for future deliberations in clinical trial design. PMID- 18510533 TI - Progress in the molecular biology of inherited bleeding disorders. PMID- 18510534 TI - Mild/moderate haemophilia A: new insights into molecular mechanisms and inhibitor development. AB - In mild/moderate haemophilia A (MHA) patients, many factor VIII (FVIII) gene defects, mainly missense mutations, have been identified and greatly improved the understanding of the structure and function of FVIII molecule. Characterization of the molecular mechanisms involved in MHA has helped to identify regions critical for proper FVIII biosynthesis, thrombin activation, intramolecular stability as well as binding regions for important intermolecular interactions with von Willebrand factor, factor IXa and the phospholipid surface. Some missense mutations were also recognized as contributing factors to inhibitor development in MHA, in parallel to acquired factors such as inflammatory state or intensity of treatment. Treatment of MHA with inhibitor patients raises questions on how best to stop or prevent bleeding episodes and eradicate the inhibitor. Longitudinal data collection is currently being conducted in France and Belgium to enhance our knowledge in this field and to further help make treatment decision. The description of mutations in MHA finally contributed to the identification of epitopes involved in the immune response to FVIII. In some patients, the epitope specificity of inhibitor antibodies recognizing normal exogenous FVIII alone and not patient ('self') FVIII was described. This distinguished epitope specificity could also be demonstrated at the T-cell clonal level. One might expect that these molecular studies will have a major impact on development of new FVIII products in the future. PMID- 18510535 TI - New images in haemophilia. AB - New imaging techniques are valuable for the care of patients with haemophilia. On angiography it is shown that some bleedings in severely damaged joints or after implantation of prostheses are arterial. Effect of clotting factor is often poor. Selective catherization with embolization of the bleeding artery stops the bleed and is clinically effective. From 31 patients with severe haemophilia A or B, 62 knee radiographs were scored according to the Pettersson-scoring system as well as with Knee Digital Image Analysis (KIDA). Using KIDA, good correlation was found for osteoporosis, irregular subchondral surface, narrowing of the joint space, deformity and incongruence. For each of the parameters within one point in the Pettersson score a large variation existed in KIDA grading. MRI is accurate for diagnosis of soft and osteochondral tissue. Nevertheless, it is costly and not very accessible. The use of parallel imaging is more feasible for assessment of multiple joints within a relatively short period of time. Although ultrasonography also holds the potential for being an adjunct to MRI it has the disadvantage that it is operator-dependent. In a cohort of 124 chronically HCV infected haemophilia patients transient elastography was performed to measure liver stiffness. 57 (46%) had no or mild fibrosis, 18 (14.5%) moderate fibrosis and 49 severe or cirrhotic fibrosis. Transient elastography is safe and helpful to refer patients to antiviral therapy. PMID- 18510536 TI - The role of the physiatrist in the haemophilia comprehensive care team in different parts of the world. AB - The role of the physiatrist, as a member of the multidisciplinary haemophilia comprehensive care team, is to prevent and treat activity limitations and restriction of participation on the part of the patient. . This role is threefold: (i) provide education to the patients, families and healthcare providers to detect disabling injuries and take adequate precautions; (ii) provide specialized treatment of musculoskeletal disabilities striving for the highest level of functionality and (iii) Stimulate the patient's full participation in socio-economic activities of his country. Prominent physiatrists from different parts of the world describe the situation in Armenia, China, Egypt, France, Indonesia (and the Philippines), the Netherlands and South America. PMID- 18510537 TI - Musculoskeletal measurement tools from the International Prophylaxis Study Group (IPSG). PMID- 18510538 TI - Orthopaedic management of haemophilia arthropathy of the ankle. AB - Joint bleeding, or haemarthrosis, is the most common type of bleeding episode experienced by individuals with haemophilia A and B. This leads to changes within the joints, including synovial proliferation, which results in further bleeding and chronic synovitis. Blood in the joint can also directly damage the cartilage, and with repeated bleeding, there is progressive destruction of both cartilage and bone. The end result is known as haemophilic arthropathy. The joints most commonly affected are the knees, elbows and ankles, although any synovial joint may be involved. In the ankle, both the tibiotalar and subtalar joints may be affected and joint bleeding and arthropathy can lead to a number of deformities. Haemophilic arthropathy can be prevented through regular factor replacement prophylaxis and implementing physiotherapy. However, when necessary, there are multiple surgical and non-surgical options available. In early ankle arthropathy with absent or minimal joint changes, both radioisotopic and chemical synoviorthesis can be used to reduce the hypertrophied synovium. These procedures can decrease the frequency of bleeding episodes, minimizing the risk of articular cartilage damage. Achilles tendon lengthening can be performed, in isolation or in combination with other surgical measures, to correct Achilles tendon contractures. Both arthroscopic and open synovectomies are available as a means to remove the friable villous layer of the synovium and are often indicated when bleeding episodes cannot be properly controlled by factor replacement therapy or synoviorthesis. In the later stages of ankle arthropathy, other surgical options may be considered. Debridement may be indicated when there are loose pieces of cartilage or anterior osteophytes, and can help to improve the joint function, even in the presence of articular cartilage damage. Supramalleolar tibial osteotomy may be indicated in patients with a valgus deformity of the hindfoot without degenerative radiographic findings. Joint fusion, or arthrodesis, is the treatment of choice in the advanced stages of ankle arthropathy although total ankle replacement is currently available. Early ankle replacement components were associated with a poor outcome, but as implant designs have improved, there have been successful outcomes achieved. As the ankle is a commonly affected joint in many individuals with haemophilia, it is important to add to the knowledge base to validate indications and timing of surgical and non-surgical interventions in ankle arthropathy. PMID- 18510539 TI - The role of synovectomy in the management of a target joint. PMID- 18510540 TI - Management of carriers and babies with haemophilia. AB - Although up to 30% of babies born with haemophilia do not have a family history of the disorder, the remaining 70% are born in families where haemophilia has been diagnosed. It has been estimated that for each male with haemophilia, there are five potential female carriers. Such women will benefit from knowledge of both their genetic (mutation present or not) and phenotype (level of plasma factor activity) status. Genetic counselling services to provide information and testing, together with plasma factor measurement, should be offered where available to all women at risk of being carriers. It is critical that women know their plasma factor measurement as they may have mild haemophilia (factor 5-30%, reference range 50-150%) which requires management at times of medical and surgical procedures and following trauma. Close liaison between adult and paediatric haemophilia centres and obstetric-gynaecology units is important to ensure that clinical carers identify and address carriers' needs. Genetic testing should be performed only after a potential carrier has been counselled and supported to receive such information. There is no coercion to accept such testing. An advantage of genetic testing is to then discuss pre-implantation genetic diagnosis which is an ex-vitro form of prenatal diagnosis. This can assist couples at risk of having a child with haemophilia who wish to reduce their anxieties about reproduction. Approximately 4% of boys with haemophilia, born in countries with good maternal care, will have intracranial haemorrhage in the neonatal period. There are no high-level evidence-based guidelines for the management of delivery or of the newborn with haemophilia. Obstetricians or other birth attendants need to be advised of the possibility of delivery of a boy with haemophilia and seek support from a haemophilia specialist during the pregnancy. The mother can then be monitored and plans for delivery be developed between her medical consultants and discussed with her. It is always preferable for a carrier to know of her genetic and phenotypic status before becoming pregnant so that she is informed as to her options and requirements for safe delivery. PMID- 18510541 TI - Haemophilia in the first years of life. AB - Surgery in infants and young children with haemophilia, when preceded by accurate diagnosis and accompanied by safe and effective factor prophylaxis, is not associated with a significant risk of haemorrhage. Haemophilic newborns undergoing circumcision or major surgery prior to diagnosis and in the absence of appropriate haemostatic prophylaxis remain as a concern. Inhibitor development has replaced haemorrhage as the major surgical complication in the developed world, largely because of the intensity of treatment used to secure haemostasis. For that reason only, essential surgery should be performed. Intracranial haemorrhage (ICH) during the neonatal period affects 3.5-4.0% of all haemophilia boys in countries with a good standard of health care, which is considerably (40 80 times) higher than expected in the normal population. Because of the high frequency of sporadic cases, ICH in the neonatal period can only be partially prevented by improved carrier diagnosis and counselling. Infections and thrombosis are the major serious complications of central venous lines. Large differences are seen in the frequency of these complications, the most plausible explanations are probably related to the protocol used for device care, the quality of education and the compliance of the users, an issue addressed in an on going study. PMID- 18510542 TI - Prophylaxis for severe haemophilia: clinical challenges in the absence as well as in the presence of inhibitors. AB - Prophylaxis is defined as the regular administration of clotting factor concentrates to prevent bleeding. Extensive data from observational studies and a recent randomized controlled trial (have established that early prophylactic treatment prevents bleeds and arthropathy in boys with severe haemophilia. The initiation of prophylaxis in young children remains challenging. To prevent arthropathy, prophylaxis should be started early, before the onset of joint damage. Alternative strategies of starting include starting before the age of 2 years, or starting before the third joint bleed. Dose and frequency vary between the original Swedish regime of 20-40 IU kg(-1) three times per week and lower dosed and step up regimes starting with 50 IU kg(-1) once weekly and rapidly increasing dose and frequency in case of bleeds. In the second decade, most patients on prophylaxis learn self-infusion. Self-management warrants confirmation of adequate knowledge of the disease. Increasing self-management concurring with major physical and psychological changes may cause reduced adherence. The challenge is to promote adherence and continue to prevent bleeds during this important period of rapid growth. The third decade of life often represents a change in lifestyle. Patients may get a job and periods of physical activity may be more confined. About two thirds of patients experiment with discontinuing prophylaxis in their early twenties, and 20-30% with mild bleeding patterns switch to on-demand treatment for prolonged periods or even permanently. The challenge is to optimize efficiency by individualizing prophylactic dose and frequency according to lifestyle and bleeding pattern. Inhibitors may develop in up to 30% of patients with severe haemophilia. Especially those with high titre inhibitors are at increased risk of developing target joints and severe arthropathy. The use of prophylactic treatment with bypassing agents in inhibitor patients is increasing. Early studies report in a significant reduction of bleeds, including intracranial bleeds, and improvement in quality of life. Data on results of primary prophylaxis in patients with inhibitors to prevent arthropathy are not yet available. PMID- 18510543 TI - Rare bleeding disorders. AB - During the haemostatic response, the formation of a primary platelet plug limits bleeding and provides a surface for clotting factors to assemble and become activated. The initial platelet plug is stabilized by fibrin monomers, covalently cross-linked by FXIII, forming a platelets-fibrin thrombus. Defects in platelets as well as inherited deficiencies of coagulation factors including fibrinogen, FII, FV, FV + FVIII, FVII, FX, FXI and FXIII deficiencies, generally lead to lifelong bleeding disorders, whose severity of bleeding symptoms is heterogeneous in platelets abnormalities but generally inversely proportional to the degree of the factor deficiency in rare bleeding disorders (RBDs). The prevalence of platelet defects among the general population has not been established, whereas for RBDs it ranges from approximately 1 in 2 million to 1 in 500,000, being higher in countries where consanguineous marriages are diffused. As a consequence of the rarity of these deficiencies, the type and severity of bleeding symptoms, the underlying molecular defects, and the actual management of bleeding episodes are not well established. In this review the main features, diagnosis, available treatment options and treatment complications of the platelet disorders, caused by abnormalities in platelet receptors for adhesive proteins, platelet receptors for soluble agonists, platelet granules, signal transduction pathways, or procoagulant phospholipids will be discussed by Dr Cattaneo, whereas fibrinogen deficiency and FXIII deficiency will be described by Dr Inbal and Dr de Moerloose, respectively. Finally, the update of the Rare Bleeding Disorders Database will be presented by Dr Spreafico. PMID- 18510544 TI - Genetic imprinting of autoantibody repertoires in systemic lupus erythematosus patients. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease distinguished by great heterogeneity in clinical manifestations and autoantibody expression. While only a handful of autoantibody specificities have proved useful for clinical diagnosis, to characterize complex lupus-associated autoantibody profiles more fully we have applied proteome microarray technology. Our multiplex microarrays included control ligands and 65-autoantigens, which represent diverse nuclear and cytoplasmic antigens recognized by disease-associated and natural autoantibodies. From longitudinal surveys of unrelated SLE patients, we found that autoantibody profile patterns can be patient-specific and highly stable overtime. From profiles of 38 SLE patients that included 14 sets of SLE twins, autoantibodies to the phospholipid neo-determinants, malondialdehyde (MDA) and phosphorylcholine (PC), which are exposed on apoptotic but not healthy cells, were among the most prevalent and highly expressed. We also found that immunoglobulin M (IgM) reactivity to MDA and PC ligands had significant direct correlations with DNA containing antigens, while such a general relationship was not found with a panel of RNA-related antigens, or for IgG-autoantibodies. Significantly, hierarchical analysis revealed co-distribution/clustering of the IgM autoantibody repertoire patterns for six of 14 twin sets, and such patterns were even more common (10 of 14) for IgG autoantibody profiles. Our findings highlight the potentially distinct roles of IgM and IgG autoantibodies, as we postulate that the direct correlations for IgM autoantibodies to DNA antigens with apoptosis-related determinants may be due to co-expression arising from common pro-homeostatic protective roles. In contrast, the sharing of IgG autoantibody fingerprints by monozygotic twins suggests that lupus IgG autoantibodies can arise in predisposed individuals in genetically determined patterns. PMID- 18510545 TI - Tietz syndrome: unique phenotype specific to mutations of MITF nuclear localization signal. PMID- 18510547 TI - X-linked hypohidrotic ectodermal dysplasia. Genetic and dental findings in 67 Danish patients from 19 families. AB - This study aimed to investigate genotype and phenotype in males affected with X linked hypohidrotic ectodermal dysplasia (HED) and in female carriers, to analyse a possible genotype-phenotype correlation, and to analyse a possible relation between severity of the symptoms and the X-chromosome inactivation pattern in female carriers. The study group comprised 67 patients from 19 families (24 affected males and 43 female carriers). All participants had clinical signs of ectodermal dysplasia and a disease-causing EDA mutation. The EDA gene was screened for mutations by single-stranded conformational polymorphism and direct sequencing. Multiplex ligation-dependent probe amplification (MLPA) analysis was used to detect deletions/duplications in female probands. Sixteen different EDA mutations were detected in the 19 families, nine not described previously. The MLPA analysis detected a deletion of exon 1 in one female proband. No genotype phenotype correlations were observed, and female carriers did not exhibit a skewed X-chromosome inactivation pattern. However, in two female carriers with pronounced clinical symptoms, in whom the parental origin of each allele was known, we observed that mainly the normal allele was inactivated. PMID- 18510546 TI - Genetic aspects of human congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a common major malformation affecting 1/3000-1/4000 births, which continues to be associated with significant perinatal mortality. Much current research is focused on elucidating the genetics and pathophysiology contributing to CDH to develop more effective therapies. The latest data suggest that many cases of CDH are genetically determined and also indicate that CDH is etiologically heterogeneous. The present review will provide a brief summary of diaphragm development and model organism work most relevant to human CDH and will primarily describe important human phenotypes associated with CDH and also provide recommendations for diagnostic evaluation of a fetus or infant with CDH. PMID- 18510548 TI - Variable phenotypes associated with 10q23 microdeletions involving the PTEN and BMPR1A genes. AB - Infantile juvenile polyposis is a rare disease with severe gastrointestinal symptoms and a grave clinical course. Recently, 10q23 microdeletions involving the PTEN and BMPR1A genes were found in four patients with infantile juvenile polyposis. It was hypothesized that a combined and synergistic effect of the deletion of both genes would explain the condition. Subsequently, however, a patient with a larger 10q23 deletion including the same genes but with a mild clinical phenotype was identified. Here, we present four additional patients with 10q23 microdeletions involving the PTEN and BMPR1A genes. The sizes of the deletions were analyzed using single nucleotide polymorphism array analysis. All patients had macrocephaly, dysmorphic features, retardation and congenital abnormalities. One patient developed colorectal cancer. However, only one case had disease onset before 2 years of age and severe symptoms requiring colectomy. No clear correlation was found between ages at onset or severity of gastrointestinal symptoms and the sizes of the deletions. We conclude that patients with 10q23 microdeletions involving the PTEN and BMPR1A genes have variable clinical phenotypes, which cannot be explained merely by the deletion sizes. The phenotypes are not restricted to severe infantile juvenile polyposis but include childhood-onset cases with macrocephaly, retardation, mild gastrointestinal symptoms and possibly early-onset colorectal cancer. PMID- 18510549 TI - The effect of L-arginine administration on muscle force and power in postmenopausal women. AB - Previously published data (J Bone Miner Res (2005); 20: 471) did not give evidence that the administration of the nitric oxide precursor L-arginine increases bone formation and decreases bone resorption in postmenopausal women. Data of this trial were reanalysed for putative effects of L-arginine on muscle mass and muscular function. Therefore, 11 females of the former study group (n=15; age 54.5+/-4.1 years; daily oral administration of 18 g L-arginine hydrochloride (equivalent of 14.2 g L-arginine) over 6 months) and 12 females of the control group (n=15; age 55.3+/-4.4 years; daily administration of 18 g dextrose over 6 months) were analysed for biomechanical parameters (MIGF, maximal isometric grip force; PJF, peak jump force; PJP, peak jump power) and for the cross-sectional muscle area (MA) and fat area (FA) at forearm and leg (calf) measured by peripheral quantitative computed tomography. The study was performed in a double-blind design. The assessment of muscular and biomechanical parameters was undertaken before and after 6 months of L-arginine versus placebo administration. L-arginine-supplemented females had a significant increase of PJF/kg in comparison with the control group. PJP/kg, MIGF, MA and FA were not significantly influenced by the administration of L-arginine. In conclusion, the administration of L-arginine increased maximal force in mechanographic analyses and may prevent a decline of muscle force in postmenopausal women. PMID- 18510550 TI - Prediction of recurrent disease by cytology and HPV testing after treatment of cervical intraepithelial neoplasia. AB - OBJECTIVE: To assess the role of human papillomavirus (HPV) testing and cytology as predictors of residual/recurrent disease after treatment of high-grade cervical intraepithelial lesions. METHODS: One hundred and thirty-eight women with cervical intraepithelial neoplasia (CIN) grade 2/3 lesion on biopsy were included in a prospective follow-up study in Belgium and Nicaragua. All women were treated with loop electrosurgical excision procedure (LEEP) and follow-up visits took place at 6 weeks, 6 months, 1 year and 2 years. During these visits, a Papanicolaou (Pap) smear test was taken, colposcopy was performed and specimens were collected for HPV testing. Cytology, high-risk (HR) HPV presence, persistent HR HPV infection and combinations of these tests at different time points during follow-up were correlated with histologically confirmed residual/recurrent disease. RESULTS: Thirteen patients (9%) developed residual/recurrent disease during follow-up. Abnormal cytology at 6 weeks after treatment was significantly correlated with residual/recurrent disease. Nine of thirty-seven patients with abnormal cytology at 6 weeks had recurrent disease versus three of seventy with a normal cytology [odds ratio (OR): 7.2; 95% confidence interval (CI): 1.8-28.5; P = 0.003). Sensitivity of this test was 75.0%, specificity 70.5%. Combining abnormal cytology and the presence of HR HPV within the first 6 months after treatment gave the best correlation with residual/recurrent disease: of the 54 women with abnormal cytology and/or HR HPV presence within the first 6 months, 11 developed residual/recurrent disease (OR 10.2; 95% CI: 2.2-48.3). Sensitivity of this combination was 84.6% and specificity 65.0%. CONCLUSION: Cytology remains the cornerstone in the early follow-up after LEEP for CIN lesions of the cervix. HPV testing can add value as it increases the sensitivity of cytology in concomitant testing within the first 6 months. PMID- 18510551 TI - Cytology and outcome of LSIL: cannot exclude HSIL compared to ASC-H. AB - OBJECTIVE: The cytological features associated with clinical outcome of 'LSIL cannot exclude HSIL (LSIL-H)' in comparison with 'atypical squamous cells cannot exclude HSIL (ASC-H)' are incompletely described. METHODS: LSIL-H and ASC-H Pap tests reported in a regional laboratory during a 13-month period were reviewed by two pathologists. Cytological features suspicious for HSIL were evaluated against a check list of 52 atypical features. All histology over 2 years of follow up for tests reclassified as LSIL-H and ASC-H was retrieved to determine clinical outcome. Atypical cytological features were correlated with outcome. RESULTS: The review yielded 89 LSIL-H and 86 ASC-H. The highest ranked atypical cytological feature in each group was increased nuclear cytoplasmic ratio. Clinical outcome was positive (CIN II/III or AIS) in 44 (49%) LSIL-H and 33 (38%) ASC-H. Round (P = 0.02) and naked nuclei (P = 0.009) were significant correlates of outcome amongst LSIL-H tests, but no feature correlated with outcome in the ASC-H group. CONCLUSIONS: LSIL-H is different to ASC-H because of the 11% higher frequency of a positive outcome and the cytological features associated with outcome. PMID- 18510552 TI - Abundant and diverse bacteria involved in DMSP degradation in marine surface waters. AB - An expanded analysis of oceanic metagenomic data indicates that the majority of prokaryotic cells in marine surface waters have the genetic capability to demethylate dimethylsulfoniopropionate (DMSP). The 1701 homologues of the DMSP demethylase gene, dmdA, identified in the (2007) Global Ocean Sampling (GOS) metagenome, are sufficient for 58% (+/-9%) of sampled cells to participate in this critical step in the marine sulfur cycle. This remarkable frequency of DMSP demethylating cells is in accordance with biogeochemical data indicating that marine phytoplankton direct up to 10% of fixed carbon to DMSP synthesis, and that most of this DMSP is subsequently degraded by bacteria via demethylation. The GOS metagenomic data also revealed a new cluster of dmdA sequences (designated Clade E) that implicates marine gammaproteobacteria in DMSP demethylation, along with previously recognized alphaproteobacterial groups Roseobacter and SAR11. Analyses of G+C content and gene order indicate that lateral gene transfer is likely responsible for the wide distribution of dmdA among diverse taxa, contributing to the homogenization of biogeochemical roles among heterotrophic marine bacterioplankton. Candidate genes for the competing bacterial degradation process that converts DMSP to the climate-active gas dimethylsulfide (DMS) (dddD and dddL) occur infrequently in the (2007) GOS metagenome, suggesting either that the key DMS-producing bacterial genes are yet to be identified or that DMS formation by free-living bacterioplankton is insignificant relative to their demethylation activity. PMID- 18510553 TI - A microdiversity study of anammox bacteria reveals a novel Candidatus Scalindua phylotype in marine oxygen minimum zones. AB - The anaerobic oxidation of ammonium (anammox) contributes significantly to the global loss of fixed nitrogen and is carried out by a deep branching monophyletic group of bacteria within the phylum Planctomycetes. Various studies have implicated anammox to be the most important process responsible for the nitrogen loss in the marine oxygen minimum zones (OMZs) with a low diversity of marine anammox bacteria. This comprehensive study investigated the anammox bacteria in the suboxic zone of the Black Sea and in three major OMZs (off Namibia, Peru and in the Arabian Sea). The diversity and population composition of anammox bacteria were investigated by both, the 16S rRNA gene sequences and the 16S-23S rRNA internal transcribed spacer (ITS). Our results showed that the anammox bacterial sequences of the investigated samples were all closely related to the Candidatus Scalindua genus. However, a greater microdiversity of marine anammox bacteria than previously assumed was observed. Both phylogenetic markers supported the classification of all sequences in two distinct anammox bacterial phylotypes: Candidatus Scalindua clades 1 and 2. Scalindua 1 could be further divided into four distinct clusters, all comprised of sequences from either the Namibian or the Peruvian OMZ. Scalindua 2 consisted of sequences from the Arabian Sea and the Peruvian OMZ and included one previously published 16S rRNA gene sequence from Lake Tanganyika and one from South China Sea sediment (97.9-99.4% sequence identity). This cluster showed only 0.05) in PT and INR determination, but there was a significant difference (P < 0.01) for APTT. The INR with local MNPT and APTTR with MNAPTT, obtained with the ACL Futura and CA 510, showed much better agreement; 98.8% (82/83) of bias for INR with local MNPT was less than 15% compared with 90.4% (75/83) of bias for INR; and 100% of bias for APTTR (62/62) was less than 15% compared with only 6.5% (4/62) of bias for APTT. Meanwhile, there was no significant difference (P = 0.865) for APTTR with MNAPTT compared with APTT (P = 0.002) between the ACL Futura and CA 510. In conclusion, these analyzers showed very poor agreement for both the PT and APTT, but the calculation of ratios significantly improved agreement. PMID- 18510576 TI - Assessment and documentation of patients' nutritional status: perceptions of registered nurses and their chief nurses. AB - AIMS: To study, within municipal care and county council care, (1) chief nurses' and registered nurses' perceptions of patient nutritional status assessment and nutritional assessment/screening tools, (2) registered nurses' perceptions of documentation in relation to nutrition and advantages and disadvantages with a documentation model. BACKGROUND: Chief nurses and registered nurses have a responsibility to identify malnourished patients and those at risk of malnutrition. DESIGN AND METHODS: In this descriptive study, 15 chief nurses in municipal care and 27 chief nurses in county council care were interviewed by telephone via a semi-structured interview guide. One hundred and thirty-one registered nurses (response rate 72%) from 14 municipalities and 28 hospital wards responded to the questionnaire, all in one county. RESULTS: According to the majority of chief nurses and registered nurses, only certain patients were assessed, on admission and/or during the stay. Nutritional assessment/screening tools and nutritional guidelines were seldom used. Most of the registered nurses documented nausea/vomiting, ability to eat and drink, diarrhoea and difficulties in chewing and swallowing, while energy intake and body mass index were rarely documented. However, the majority documented their judgement about the patient's nutritional condition. The registered nurses perceived the VIPS model (Swedish nursing documentation model) as a guideline as well as a model obstructing the information exchange. Differences were found between nurses (chief nurses/registered nurses) in municipal care and county council care, but not between registered nurses and their chief nurses. CONCLUSIONS: All patients are not nutritionally assessed and important nutritional parameters are not documented. Nutritionally compromised patients may remain unidentified and not properly cared for. RELEVANCE TO CLINICAL PRACTICE: Assessment and documentation of the patients' nutritional status should be routinely performed in a more structured way in both municipal care and county council care. There is a need for increased nutritional nursing knowledge. PMID- 18510577 TI - Genetic engineering of improved nitrogen use efficiency in rice by the tissue specific expression of alanine aminotransferase. AB - Summary Nitrogen is quantitatively the most essential nutrient for plants and a major factor limiting crop productivity. One of the critical steps limiting the efficient use of nitrogen is the ability of plants to acquire it from applied fertilizer. Therefore, the development of crop plants that absorb and use nitrogen more efficiently has been a long-term goal of agricultural research. In an attempt to develop nitrogen-efficient plants, rice (Oryza sativa L.) was genetically engineered by introducing a barley AlaAT (alanine aminotransferase) cDNA driven by a rice tissue-specific promoter (OsAnt1). This modification increased the biomass and grain yield significantly in comparison with control plants when plants were well supplied with nitrogen. Compared with controls, transgenic rice plants also demonstrated significant changes in key metabolites and total nitrogen content, indicating increased nitrogen uptake efficiency. The development of crop plants that take up and assimilate nitrogen more efficiently would not only improve the use of nitrogen fertilizers, resulting in lower production costs, but would also have significant environmental benefits. These results are discussed in terms of their relevance to the development of strategies to engineer enhanced nitrogen use efficiency in crop plants. PMID- 18510578 TI - Analysis of enzyme-treated red blood cell surface and haemagglutination using a theory of soft-particle electrophoresis. AB - BACKGROUND AND OBJECTIVES: Enzymatic treatment of red blood cells is thought to reduce the cell zeta (zeta) potential, effectively decreasing the distance between cells to less than the length of an immunoglobulin G antibody binding site, and resulting in agglutination of cells. However, the zeta potential given by Smoluchowski's formula is based on theories of the electrophoresis of hard colloidal particles. A theory has recently been developed for the electrophoresis of colloidal particles covered with polyelectrolytes, which we call 'soft particles'. MATERIALS AND METHODS: The electrophoretic mobility of red blood cell treated with papain and neuraminidase was measured as the electrolyte concentration of the medium using phosphate buffer. The results were analysed via the formula for 'soft particles'. This mobility formula involved two parameters, the fixed charge density (ZN) and parameter 1/lambda characterizing the 'softness' of the cell surface layer. RESULTS: The best-fit curves of 0.1 units neuraminidase-treated red blood cells indicated that ZN decreased by 76% and 1/lambda decreased by 8% compared to intact red blood cells. In contrast, in 5 units of papain-treated red blood cells ZN decreased by 45% and 1/lambda decreased by 33% compared to intact red blood cells. CONCLUSION: The present study shows that the change in ZN for neuraminidase-treated cells was very large, but the cells did not become agglutinable. Papain-treated cells had changes in both ZN and 1/lambda, and the cells became agglutinable. 1/lambda is one of the important factors for agglutination. PMID- 18510579 TI - Introduction of a real-time-based blood-group genotyping approach. AB - BACKGROUND AND OBJECTIVE: Genotyping may be applied for rare blood group polymorphisms in a high-throughput mode as well for the molecular determination of blood groups due to unclear serological results. MATERIAL AND METHODS: We developed and validated a DNA typing method for the determination of KEL1/2, JK1/2, FY1/2, FY0, MNS1/2, MNS3/4, DO1/2, CO1/2 and LU1/2 alleles using a melting curve analysis downstream from a fully automated DNA extraction. All assays were validated in terms of specificity, sensitivity, assay variability and robustness. The usability was proven by a batch of 200 blood samples with partially known phenotype. RESULTS: Assays for all blood groups were within the range of specificity (100%), assay variability and robustness (coefficient of variance < 3%). Genotypes of 200 random platelet donors were fully consistent with existing phenotype data. The obtained genotype distribution is in complete concordance with existing data for the European population underlined by a complete absence of CO2 homozygous donors and the FY0 allele among the cohort. CONCLUSION: We introduce an approach for blood group genotyping of particular samples or gene loci in glass capillary format and for medium-throughput analysis in 96/384-well format. The advantages of this real-time polymerase chain reaction method are its automation potential, the flexibility regarding hardware and the rapid cycling time. PMID- 18510580 TI - Validation of a haemoglobin dilution method for estimation of blood loss. AB - BACKGROUND AND OBJECTIVES: Analysis of haemoglobin (Hb) dilution after bleeding is a simple, inexpensive and non-invasive method to estimate blood loss. Blood volume is estimated, taking sex, weight and height into account. The Hb concentration before and after blood loss is analysed and, from the difference, the blood loss volume can be calculated assuming a normovolemic subject. Although widely used this method has never been validated. MATERIAL AND METHODS: The Hb concentration of 21 blood donors was analysed before and up to 4 days after a standard blood donation and in another 18 blood donors the Hb concentration was analysed before and on day 4, 6, 8, 11 and 14 after blood donation. The blood volume of each donor was calculated and the donated blood volume was estimated by weighing. We calculated the blood loss by the Hb dilution method and compared the calculated value with the donated blood volume. RESULTS: The mean donated blood volume was 442 +/- 10 ml, whereas the mean calculated blood loss was 152 +/- 214 ml using the Hb concentration of the first day after donation and 301 +/- 145 ml with the Hb concentration of day 6 after blood donation after which no further Hb decrease was observed. The directly measured Hb concentration was always higher than the calculated/expected Hb concentration based on the blood donation volume. CONCLUSIONS: The Hb dilution method underestimates the true blood loss by more than 30% after a moderate blood loss of approximately 10% of the total blood volume. PMID- 18510581 TI - Asthma pharmacotherapy prescribing in the ambulatory population of the United States: evidence of nonadherence to national guidelines and implications for elderly people. AB - OBJECTIVES: To examine the level of physician adherence to the Expert Panel Report 2 (EPR-2) pharmacotherapy guidelines of the asthma population, specifically in the elderly ambulatory patient population of the United States. DESIGN: Retrospective cross-sectional study using a national survey. SETTING: National Ambulatory Medical Care Survey data of U.S. elderly patients from 1998 through 2004. PARTICIPANTS: The weighted population sample size was 82,020,318 patients. There were 1,540 observations in this study (preweighted sample size) and 96 strata, with 446 population sampling units (PSUs). There were 11,868,340 patients that were elderly, and they accounted for 14.5% of the overall population sampled. MEASUREMENTS: Specific patient demographic variables, physician demographic variables, and information about asthma medications prescribed were extracted from the data set and analyzed. Descriptive statistics for the patient demographic, physician demographic, and asthma pharmacotherapy variables were generated. A series of logistic regression models were created, with the choice of asthma pharmacotherapy agent used as the dependent variable and patient and physician demographic variables as the independent variables. RESULTS: A major finding was that physicians were not adherent to the National Asthma Education and Prevention Program EPR-2 asthma pharmacotherapy guidelines. Another finding was that, although elderly patients (aged >or=65) were exposed to more-stable patterns of care, they were less likely to be prescribed controller medications, long-acting bronchodilators (LABAs), combinations of inhaled corticosteroids and LABAs, and short-acting beta agonists than patients aged 35 to 64. CONCLUSION: A more-concerted effort needs to be undertaken to improve physician adherence to the EPR-2 guidelines, especially in prescribing asthma pharmacotherapy to elderly patients. PMID- 18510582 TI - Recurrent falls and dual task-related decrease in walking speed: is there a relationship? AB - OBJECTIVES: To determine whether dual task-related changes in walking speed were associated with recurrent falls in frail older adults. DESIGN: Twelve-month prospective cohort study. SETTING: Thirteen senior housing facilities. PARTICIPANTS: Two hundred thirteen subjects (mean age 84.4+/-5.5). MEASUREMENTS: Usual and dual-tasking walking speeds (m/s) were calculated on a 10-m straight walkway at baseline. Information on incident falls during the follow-up year was collected monthly, and participants were divided into three groups based on the occurrence of falls (0, 1, and >or=2). Recurrent falls were defined as two or more falls during the 12-month follow-up period. RESULTS: Twenty subjects (9.4%) were classified as recurrent fallers. The occurrence of recurrent falls was associated with age (crude odds ratio (OR)=1.11, P=.02), number of drugs (crude OR=1.28, P=.002), and walking speed under both walking conditions (crude OR=0.96, P=.002 for usual walking and crude OR=0.60, P=.005 for walking while counting backward). Multiple Poisson regression showed that only walking speed while dual tasking and number of drugs were associated with incident falls (incident rate ratio (IRR)=0.84, P=.045 and IRR=1.10, P=.004). CONCLUSION: Slower walking speed while counting backward was associated with recurrent falls, suggesting that changes in gait performance while dual tasking might be an inexpensive way of identifying frail older adults prone to falling. PMID- 18510583 TI - Anticholinergic activity of 107 medications commonly used by older adults. AB - The objective of this study was to measure the anticholinergic activity (AA) of medications commonly used by older adults. A radioreceptor assay was used to investigate the AA of 107 medications. Six clinically relevant concentrations were assessed for each medication. Rodent forebrain and striatum homogenate was used with tritiated quinuclidinyl benzilate. Drug-free serum was added to medication and atropine standard-curve samples. For medications that showed detectable AA, average steady-state peak plasma and serum concentrations (C(max)) in older adults were used to estimate relationships between in vitro dose and AA. All results are reported in pmol/mL of atropine equivalents. At typical doses administered to older adults, amitriptyline, atropine, clozapine, dicyclomine, doxepin, L-hyoscyamine, thioridazine, and tolterodine demonstrated AA exceeding 15 pmol/mL. Chlorpromazine, diphenhydramine, nortriptyline, olanzapine, oxybutynin, and paroxetine had AA values of 5 to 15 pmol/mL. Citalopram, escitalopram, fluoxetine, lithium, mirtazapine, quetiapine, ranitidine, and temazepam had values less than 5 pmol/mL. Amoxicillin, celecoxib, cephalexin, diazepam, digoxin, diphenoxylate, donepezil, duloxetine, fentanyl, furosemide, hydrocodone, lansoprazole, levofloxacin, metformin, phenytoin, propoxyphene, and topiramate demonstrated AA only at the highest concentrations tested (patients with above-average C(max) values, who receive higher doses, or are frail may show AA). The remainder of the medications investigated did not demonstrate any AA at the concentrations examined. Psychotropic medications were particularly likely to demonstrate AA. Each of the drug classifications investigated (e.g., antipsychotic, cardiovascular) had at least one medication that demonstrated AA at therapeutic doses. Clinicians can use this information when choosing between equally efficacious medications, as well as in assessing overall anticholinergic burden. PMID- 18510584 TI - Molecular evolution and population genetics of two Drosophila mettleri cytochrome P450 genes involved in host plant utilization. AB - Understanding the genetic basis of adaptation is one of the primary goals of evolutionary biology. The evolution of xenobiotic resistance in insects has proven to be an especially suitable arena for studying the genetics of adaptation, and resistant phenotypes are known to result from both coding and regulatory changes. In this study, we examine the evolutionary history and population genetics of two Drosophila mettleri cytochrome P450 genes that are putatively involved in the detoxification of alkaloids present in two of its cactus hosts: saguaro (Carnegiea gigantea) and senita (Lophocereus schottii). Previous studies demonstrated that Cyp28A1 was highly up-regulated following exposure to rotting senita tissue while Cyp4D10 was highly up-regulated following exposure to rotting saguaro tissue. Here, we show that a subset of sites in Cyp28A1 experienced adaptive evolution specifically in the D. mettleri lineage. Moreover, neutrality tests in several populations were also consistent with a history of selection on Cyp28A1. In contrast, we did not find evidence for positive selection on Cyp4D10, although this certainly does not preclude its involvement in host plant use. A surprising result that emerged from our population genetic analyses was the presence of significant genetic differentiation between flies collected from different host plant species (saguaro and senita) at Organ Pipe National Monument, Arizona, USA. This preliminary evidence suggests that D. mettleri may have evolved into distinctive host races that specialize on different hosts, a possibility that warrants further investigation. PMID- 18510585 TI - Phylogeography and local endemism of the native Mediterranean brine shrimp Artemia salina (Branchiopoda: Anostraca). AB - There has been a recent appreciation of the ecological impacts of zooplanktonic species invasions. The North American brine shrimp Artemia franciscana is one such alien invader in hyper-saline water ecosystems at a global scale. It has been shown to outcompete native Artemia species, leading to their local extinction. We used partial sequences of the mitochondrial Cytochrome c Oxidase Subunit 1 (COI or cox1) gene to investigate the genetic diversity and phylogeography of A. salina, an extreme halophilic sexual brine shrimp, over its known distribution range (Mediterranean Basin and South Africa) and to assess the extent of local endemism, the degree of population structure and the potential impact of traditional human saltpan management on this species. We also examined the phylogenetic relationships in the genus Artemia using COI sequences. Our results show extensive regional endemism and indicate an early Pleistocene expansion of A. salina in the Mediterranean Basin. Subsequent population isolation in a mosaic of Pleistocene refugia is suggested, with two or three refugia located in the Iberian Peninsula. Two instances of long-distance colonization were also observed. Surprisingly, given its strong phylogeographical structure, A. salina showed a signature of correlation between geographical and genetic distance. Owing to strong 'priority effects', extensive population differentiation is retained, despite dispersal via migrant birds and human management of saltpans. The foreseeable expansion of A. franciscana is likely to be followed by substantial loss of genetic diversity in Mediterranean A. salina. Large genetic divergences between Mediterranean and South African A. salina suggest that the latter deserves species status. PMID- 18510586 TI - Inferring recruitment history from spatial genetic structure within populations of the colonizing tree Albizia julibrissin (Fabaceae). AB - Comparative analyses of spatial genetic structure (SGS) among species, populations, or cohorts give insight into the genetic consequences of seed dispersal in plants. We analysed SGS of a weedy tree in populations with known and unknown recruitment histories to first establish patterns in populations with single vs. multiple founders, and then to infer possible recruitment scenarios in populations with unknown histories. We analysed SGS in six populations of the colonizing tree Albizia julibrissin Durazz. (Fabaceae) in Athens, Georgia. Study sites included two large populations with multiple, known founders, two small populations with a single, known founder, and two large populations with unknown recruitment histories. Eleven allozyme loci were used to genotype 1385 individuals. Insights about the effects of colonization history from the SGS analyses were obtained from correlograms and Sp statistics. Distinct differences in patterns of SGS were identified between populations with multiple founders vs. a single founder. We observed significant, positive SGS, which decayed with increasing distance in the populations with multiple colonists, but little to no SGS in populations founded by one colonist. Because relatedness among individuals is estimated relative to a local reference population, which usually consists of those individuals sampled in the study population, SGS in populations with high background relatedness, such as those with a single founder, may be obscured. We performed additional analyses using a regional reference population and, in populations with a single founder, detected significant, positive SGS at all distances, indicating that these populations consist of highly related descendants and receive little seed immigration. Subsequent analyses of SGS in size cohorts in the four large study populations showed significant SGS in both juveniles and adults, probably because of a relative lack of intraspecific demographic thinning. SGS in populations of this colonizing tree is pronounced and persistent and is determined by the number and relatedness of founding individuals and adjacent seed sources. Patterns of SGS in populations with known histories may be used to indirectly infer possible colonization scenarios for populations where it is unknown. PMID- 18510587 TI - Sequence differentiation in regions identified by a genome scan for local adaptation. AB - Genome scans using large numbers of randomly selected markers have revealed a small proportion of loci that deviate from neutral expectations and so may mark genomic regions that contribute to local adaptation. Measurements of sequence differentiation and identification of genes in these regions is important but difficult, especially in organisms with limited genetic information available. We have followed up a genome scan in the marine gastropod, Littorina saxatilis, by searching a bacterial artificial chromosome library with differentiated and undifferentiated markers, sequencing four bacterial artificial chromosomes and then analysing sequence variation in population samples for fragments at, and close to the original marker polymorphisms. We show that sequence differentiation follows the patterns expected from the original marker frequencies, that differentiated markers identify independent and highly localized sites and that these sites fall outside coding regions. Two differentiated loci are characterized by insertions of putative transposable elements that appear to have increased in frequency recently and which might influence expression of downstream genes. These results provide strong candidate loci for the study of local adaptation in Littorina. They demonstrate an approach that can be applied to follow up genome scans in other taxa and they show that the genome scan approach can lead rapidly to candidate genes in nonmodel organisms. PMID- 18510588 TI - SKI is critical for repressing the growth inhibitory function of TGF-beta in human melanoma. PMID- 18510589 TI - Dose-dependent, complete response to imatinib of a metastatic mucosal melanoma with a K642E KIT mutation. PMID- 18510590 TI - The complex role of vitamin D in autoimmune diseases. AB - Vitamin D, besides having well-known control functions of calcium and phosphorus metabolism, bone formation and mineralization, also has a role in the maintenance of immune-homeostasis. The immune-regulatory role of vitamin D affects both the innate and adaptive immune system contributing to the immune-tolerance of self structures. Impaired vitamin D supply/regulation, amongst other factors, leads to the development of autoimmune processes in animal models of various autoimmune diseases. The administration of vitamin D in these animals leads to improvement of immune-mediated symptoms. Moreover, in human autoimmune diseases, such as multiple sclerosis, or rheumatoid arthritis the pathogenic role of vitamin D has been described. The review aims at describing the complex immune-regulatory role of vitamin D from the cellular level through autoimmune animal models and depicting the known contribution of vitamin D in the pathogenesis of human autoimmune diseases. PMID- 18510591 TI - The importance of 'Mechnikov's thorn' for an improved understanding of 21st century medicine and immunology: a view from the eye. AB - In 1908, Ehrlich and Mechnikov shared the Nobel Prize in Medicine for their independent studies that set the scene for the modern understanding of innate and adaptive immunity. However, 20th century immunology thinking was dominated by aberrant adaptive immunity but this never adequately explained the full spectrum of inflammatory disease. This article draws on medical observations, from where immunology originated, and uses the example of the eye to illustrate how the integration of medicine and immunology leads to an improved understanding of inflammation against self. The spectrum of ocular inflammation can be viewed as either predominantly adaptive immune mediated (mostly the realm of immunology), or predominantly due to ocular tissues factors that lead to regional innate immune activation (the realm of medicine), or a variable interaction between the two. Just as the thorns that Mechnikov inserted into molluscs lead to localized innate immune activation; ocular inflammation can likewise be driven by non immune factors that include tissue degeneration or microdamage. The present article emphasizes the importance of such factors in the initiation or phenotypic expression of ocular immunopathology allowing different immunological dogmas including self-non-self discrimination, immunological tolerance and immunoprivilege to be viewed in a different light. This scheme also leads to an appreciation of how the innate immune system may be the sole perpetuator of some ocular immunopathologies. We propose that this integrated view of medicine and immunology is crucial for understanding immunology from a translational angle and has implications far beyond ocular disease. PMID- 18510592 TI - Performance-based functional evaluation of non-operative and operative treatment after anterior cruciate ligament injury. AB - Highly active anterior cruciate ligament (ACL)-injured patients are usually recommended surgical treatment as the primary intervention. The objective of this study was to compare the functional outcome in a cohort of individuals after non operative treatment to individuals after surgical treatment at a 1-year follow up. One hundred and twenty-five subjects with a mean age of 27.2 years (+/-8.6 years), and participating in level I or II activities were included. Baseline and 1-year follow-up examination included four single-legged hop tests, IKDC 2000, KOS-ADLS, KT-1000 knee arthrometer measurement, VAS, episodes of giving way, and activity level. Fifty-one percent went through non-operative treatment. Non operated subjects performed significantly better on two of the four single-legged hop tests compared with the ACL-reconstructed subjects at the 1-year follow-up. No other differences were observed. Both groups performed an average >90% compared with their uninjured leg on all single-legged hop tests at the 1-year follow-up. The IKCD 2000 scores in the non-operated and ACL-reconstructed group were on average 86 and 87. ACL-injured subjects should be informed of the possibility of success after non-operative treatment, but future studies are needed to determine significant predictive factors for success for non-operative and surgically treated individuals. PMID- 18510593 TI - The impact of fluctuations in boat velocity during the rowing cycle on race time. AB - In competitive rowing, the fluctuations in boat velocity during the rowing cycle are associated with an increased water resistance of the boat as compared with a boat moving at a constant velocity. We aimed to quantify the influence of the increased water resistance on race time using a mathematical approximation, based on the increase in physiological power being proportional to the 2nd power of boat speed. Biomechanical data (oar force, rowing angle, boat velocity, and boat acceleration) were measured when eight elite coxless pair crews performed a rowing test with a stepwise increasing stroke rate (SR: 20, 24, 28, and 32 min( 1)) that successively increased the mean boat speed. The results revealed a +4.59 s (SR 24.2) to +5.05 s (SR 31.5) 2000-m race-time difference compared with a boat hypothetically moving without velocity fluctuations. Velocity fluctuations were highly correlated with SR (r=0.93) because the accelerations of the rowers' body mass and the mass of the counteracting boat increase with SR. The possibilities to reduce velocity fluctuations and therefore race time are limited. For elite rowers, race time may be slightly reduced by a moderate reduction in SR that is compensated by an increased force output for each stroke. PMID- 18510594 TI - Serial assessments to determine normalization of gait following anterior cruciate ligament reconstruction. AB - There is an increased emphasis to identify clinically applicable methods that quantify gait deficits following anterior cruciate ligament reconstruction (ACLR). The purpose of this study was to perform serial gait assessments in a clinical setting to determine whether and when clinical gait parameters normalize in patients following ACLR. The hypothesis was that a clinically available gait treadmill would quantify gait deficits measured at 4 weeks post-reconstruction. The secondary hypothesis was that patients would demonstrate incremental improvements in these gait parameters measured at each interval up to 12 weeks post-reconstruction, and that the objectively measured improvements would correlate to the patient's subjective rating of function. Fifteen subjects, five male and 10 female, who had initial unilateral anterior cruciate ligament injury were selected for this study on the basis of operative data. All subjects were evaluated in a physical therapy clinic within 3 days following ACLR and were enrolled in a standardized rehabilitation program. The dependent gait variables of step length, stance time and gait velocity were measured at 4, 6, 8 and 12 weeks post-ACLR on a commercially available gait treadmill. A 2 x 4 multivariate analysis of variance (2 within factors) with measures for limb involvement (uninvolved and involved) and repeated measures for time (4, 6, 8 and 12 weeks) was used to assess the interactions and the main effects on the gait variables of stance time and step length. The results of this study supported the hypothesis that gait deficits and serial improvements can be objectively quantified in a clinical setting (P<0.001). Specifically, stance time, step length and gait velocity deficits evaluated at 4 weeks showed significant improvements at the measured intervals. Step length normalized at week 8. Stance time and gait velocity reached normal levels at the 12-week time interval. Subjective activity of daily living scores (ADLS) also improved following the 12-week rehabilitation, from 53+/-17% to a mean score of 88+/-11% (P<0.001). ADLS scores significantly correlated to step length (R=0.63) and stance time (R=0.53) in the involved limb. Self-selected gait velocity also correlated to ADLS scores and significantly predicted 49% of the variance in the subjective outcome measure. A clinically available gait treadmill can be used to quantify gait deficits and improvements following ACLR. Serial assessments of walking gait may aid clinicians to identify and target deficits in their patients during rehabilitation. PMID- 18510595 TI - Golf: a game of life and death--reduced mortality in Swedish golf players. AB - The specific health benefits achieved from different forms and patterns of leisure-time physical activity are not established. We analyzed the mortality in a cohort of Swedish golf players. We used the Swedish Golf Federation's membership registry and the nationwide Mortality Registry. We calculated standardized mortality ratios (SMR) with stratification for age, sex, and socioeconomic status. The cohort included 300 818 golfers, and the total number of deaths was 1053. The overall SMR was 0.60 [95% confidence intervals (CIs): 0.57-0.64]. The mortality reduction was observed in men and women, in all age groups, and in all socioeconomic categories. Golfers with the lowest handicap (the most skilled players) had the lowest mortality; SMR=0.53 (95% CI: 0.41-0.67) compared with 0.68 (95% CI: 0.61-0.75) for those with the highest handicap. While we cannot conclude with certainty that all the 40% decreased mortality rates are explained by the physical activity associated with playing golf, we conclude that most likely this is part of the explanation. To put the observed mortality reduction in context, it may be noted that a 40% reduction of mortality rates corresponds to an increase in life expectancy of about 5 years. PMID- 18510596 TI - The effect of molecular adsorbent recirculating system treatment on survival, native liver recovery, and need for liver transplantation in acute liver failure patients. AB - Acute liver failure (ALF) is a medical emergency. Molecular adsorbent recirculating system (MARS), an artificial liver support system, can partly compensate for the detoxifying function of the liver by removing toxins from blood. To analyze the efficacy of MARS treatment, the outcomes of 113 ALF patients, treated with MARS between 2001 and 2007, were compared with a historical control group of 46 ALF patients treated without MARS between 1995 and 2001. Overall survival of transplanted patients was 94% in the MARS group and 77% in the control group (P=0.06). Without transplantation, survival was 66% and 40% (P=0.03), respectively. However, the etiological distribution of ALF differed significantly between the groups. In ALF patients with unknown etiology, groups were comparable at baseline; 91% and 69% of transplanted patients survived the MARS and control groups and the native liver recovered in 20% and 8% of the patients, respectively. Of the originally nonencephalopathic patients of unknown etiology, 36% underwent liver transplantation in the MARS group compared to 100% in the control group. Interpretation of the results was difficult in toxic etiology patients on account of differing baseline statuses. MARS treatment might partly explain the trend toward increased survival of ALF patients with unknown etiology. PMID- 18510598 TI - White matter lesion severity is associated with reduced cognitive performances in patients with normal CSF Abeta42 levels. AB - OBJECTIVE: To identify possible associations between white matter lesions (WML) and cognition in patients with memory complaints, stratified in groups with normal and low cerebrospinal fluid (CSF) Abeta42 values. MATERIAL AND METHODS: 215 consecutive patients with subjective memory complaints were retrospectively included. Patients were stratified into two groups with normal (n = 127) or low (n = 88) CSF Abeta42 levels (cut-off is 450 ng/l). Cognitive scores from the Mini Mental State Examination (MMSE) and the Neurobehavioral Cognitive Status Examination (Cognistat) were used as continuous dependent variables in linear regression. WML load was used as a continuous independent variable and was scored with a visual rating scale. The regression model was corrected for possible confounding factors. RESULTS: WML were significantly associated with MMSE and all Cognistat subscores except language (repetition and naming) and attention in patients with normal CSF Abeta42 levels. No significant associations were observed in patients with low CSF Abeta42. CONCLUSIONS: WML were associated with affection of multiple cognitive domains, including delayed recall and executive functions, in patients with normal CSF Abeta42 levels. The lack of such associations for patients with low CSF Abeta42 (i.e. with evidence for amyloid deposition), suggests that amyloid pathology may obscure cognitive effects of WML. PMID- 18510599 TI - Anaphylactic reaction to omeprazole. PMID- 18510600 TI - Observations of the Indian enigma is valid. PMID- 18510601 TI - Tuberculous abdominal cocoon: a case report and review of the literature. PMID- 18510602 TI - The relationship between obesity and GERD: "big or overblown". AB - The rates of obesity have doubled over the last 22 yr in the United States. Similarly, the prevalence of gastroesophageal reflux disease (GERD) has also been rising in the United States, with studies suggesting it may now be as high as 1 in 5. Given these parallel time trends, it is tempting to speculate that obesity may, in some way, promote the development of GERD. Of the proposed mechanisms, the most attractive ones focus on the mechanical stresses imposed on the antireflux barrier, specifically increased pressure gradients across the esophagogastric junction (EGJ). In this issue of the American Journal of Gastroenterology, de Vries et al. provide a comprehensive retrospective analysis of the relationship among body mass index (BMI), pressure gradients across the EGJ, and objective evidence of GERD. While their results support that increased BMI is associated with increased pressure gradients through the EGJ and the propensity to develop hiatus hernia, they do not illustrate a direct relationship between increased gastroesophageal pressure gradients and abnormal acid exposure in the esophagus. These results highlight the complexity of this relationship and once more support the concept that GERD is a multifactorial disease that cannot be defined by a single anatomical or functional abnormality. PMID- 18510603 TI - Gastroesophageal pressure gradients in gastroesophageal reflux disease: relations with hiatal hernia, body mass index, and esophageal acid exposure. AB - OBJECTIVES: The roles of intragastric pressure (IGP), intraesophageal pressure (IEP), gastroesophageal pressure gradient (GEPG), and body mass index (BMI) in the pathophysiology of gastroesophageal reflux disease (GERD) and hiatal hernia (HH) are only partly understood. METHODS: In total, 149 GERD patients underwent stationary esophageal manometry, 24-h pH-metry, and endoscopy. RESULTS: One hundred three patients had HH. Linear regression analysis showed that each kilogram per square meter of BMI caused a 0.047-kPa increase in inspiratory IGP (95% confidence interval [CI] 0.026-0.067) and a 0.031-kPa increase in inspiratory GEPG (95% CI 0.007-0.055). Each kilogram per square meter of BMI caused expiratory IGP to increase with 0.043 kPa (95% CI 0.025-0.060) and expiratory IEP with 0.052 kPa (95% CI 0.027-0.077). Each added year of age caused inspiratory IEP to decrease by 0.008 kPa (95% CI -0.015-0.001) and inspiratory GEPG to increase by 0.008 kPa (95% CI 0.000-0.015). In binary logistic regression analysis, HH was predicted by inspiratory and expiratory IGP (odds ratio [OR] 2.93 and 2.62, respectively), inspiratory and expiratory GEPG (OR 3.19 and 2.68, respectively), and BMI (OR 1.72/5 kg/m(2)). In linear regression analysis, HH caused an average 5.09% increase in supine acid exposure (95% CI 0.96-9.22) and an average 3.46% increase in total acid exposure (95% CI 0.82-6.09). Each added year of age caused an average 0.10% increase in upright acid exposure and a 0.09% increase in total acid exposure (95% CI 0.00-0.20 and 0.00-0.18). CONCLUSIONS: BMI predicts IGP, inspiratory GEPG, and expiratory IEP. Age predicts inspiratory IEP and GEPG. Presence of HH is predicted by IGP, GEPG, and BMI. GEPG is not associated with acid exposure. PMID- 18510604 TI - Bile acid-stimulated expression of the farnesoid X receptor enhances the immune response in Barrett esophagus. AB - OBJECTIVES: Barrett's esophagus (BE) is a premalignant condition of the esophagus. It is a consequence of mucosal injury from chronic gastroesophageal reflux in which bile acids are an important toxic component. The farnesoid X receptor (FXR) is a nuclear receptor involved in the regulation of bile acid synthesis, transport, and absorption. FXR activation is also involved in the induction of the innate immune response. This suggests that FXR is involved in the pathogenesis and the inflammation seen in BE. METHODS: mRNA levels of FXR and the FXR-regulated genes, ileal bile acid-binding protein (IBABP), small heterodimer partner (SHP), and chemokines interleukin (IL)-8 and macrophage inflammatory protein 3 alpha (MIP3 alpha), were determined by real time polymerase chain reaction (RT-PCR). Protein expression was determined by immunohistochemistry. RESULTS: FXR was not expressed in squamous epithelium of healthy subjects (N = 7), but was present in both squamous and columnar epithelium of BE patients. Compared to the squamous epithelium of BE patients, their columnar epithelium displayed a 2.3-fold (P= 0.02) increase in FXR mRNA. Also, IBABP (2.2-fold; P= 0.0029), SHP (2.7-fold; P= 0.007), IL-8 (1.5-fold; P= 0.04), and MIP3 alpha (1.7-fold; P= 0.019) transcription levels were increased. Exposure of esophageal cell line TE7 to deoxycholic acid (DCA) resulted in a similar induction. The induction was abolished by the FXR antagonist guggulsterone. CONCLUSIONS: Expression levels of the bile acid receptor FXR, the bile acid metabolism genes IBABP and SHP, and the chemokines IL-8 and MIP3 alpha are increased in Barrett's epithelium. The in vitro induction of FXR by DCA suggests that bile acids can actively induce the inflammatory response in BE by recruiting immune cells. PMID- 18510605 TI - Surgical efficiency or eradication sufficiency. AB - The management of local and early mucosal cancer of the esophagus is an enormous challenge. There is a change in the practice developing with the establishment of endoscopic mucosal resection and other ablative techniques. Das et al. in this issue have used the Surveillance Epidemiology and End Results (SEER) database of the National Cancer Institute data to compare survival of 99 patients treated using endotherapy (endoscopic mucosal resection, thermal ablation, and photodynamic therapy), with 742 patients treated with surgical resection. Using the Cox proportional hazards model, there was no survival difference between the groups. The survival predictors were the age at diagnosis and the absence of radiotherapy. Although many patients are unsuitable for surgical excision and are offered alternative therapy, these data provide evidence that endotherapy may be an equally effective alternative for many patients with early esophageal cancer. It supports the need for randomized data where this can be achieved. These population-based data open up the possibility of wider acceptance of endoscopic techniques. PMID- 18510606 TI - A comparison of endoscopic treatment and surgery in early esophageal cancer: an analysis of surveillance epidemiology and end results data. AB - OBJECTIVES: Endoscopic therapy for early esophageal cancer is gaining gradual acceptance in the United States. However, little information is available regarding long-term outcome of endoscopic therapy compared to surgical treatment of early esophageal cancer. We aimed to analyze outcomes in terms of cancer-free survival in patients with early esophageal cancer managed with either endoscopic therapy or surgical resection. METHODS: The Surveillance Epidemiology and End Results database of the National Cancer Institute was searched to identify all patients who were diagnosed with stage 0 and stage 1 nonsquamous and squamous cell-type esophageal cancer between 1998 and 2003. Data on demographic features, tumor characteristics, types of treatment received (endoscopic vs surgical resection), and esophageal cancer-specific mortality were analyzed. RESULTS: Data were available for analysis in 742 patients with early esophageal cancer. Only 99 (13.3%) of these underwent endoscopic treatment (group A). The remainder of the patients was managed by surgical resection (group B). In the Cox proportional hazards model, the relative hazard for esophageal cancer-specific mortality in group A was not different from that of group B (relative hazard [RH] 0.89, 95% confidence interval [CI] 0.51-1.56, P= 0.68). The significant predictors of survival were age at diagnosis (RH 1.06, 95% CI 1.03-1.08, P < 0.001) and absence of exposure to radiation therapy (RH 0.32, 95% CI 0.21-0.48, P < 0.001). CONCLUSIONS: Patients with early esophageal cancer managed with endoscopic therapy have equivalent long-term survival compared to those treated with surgical resection. These are the first population-based data that support the effectiveness of endoscopic therapy for managing these patients. PMID- 18510607 TI - The effect of hypnosis on systemic and rectal mucosal measures of inflammation in ulcerative colitis. AB - OBJECTIVES: Hypnotherapy is effective in several diseases with a psychosomatic component. Our aim was to study the effects of one session of hypnosis on the systemic and rectal mucosal inflammatory responses in patients with active ulcerative colitis (UC). METHODS: In total, 17 patients with active UC underwent a 50-min session of gut-focused hypnotherapy. Before and after each procedure, the systemic inflammatory response was assessed by serum interleukin (IL)-6 and IL-13 concentrations, tumor necrosis factor-alpha (TNF-alpha) and IL-6 production by lipopolysaccharide (LPS)-stimulated whole blood, leukocyte count, natural killer (NK) cell number, platelet activation, and platelet-leukocyte aggregate formation. Rectal inflammation was assessed by mucosal release of substance P (SP), histamine, IL-13 and TNF-alpha, reactive oxygen metabolite production, and mucosal blood flow. Eight patients with active UC underwent a control procedure. RESULTS: Hypnosis decreased pulse by a median 7 beats per minute (bpm) (P= 0.0008); it also reduced the median serum IL-6 concentration by 53% (P= 0.001), but had no effect on the other systemic variables assessed. Hypnosis reduced rectal mucosal release of SP by a median 81% (P= 0.001), histamine by 35% (P= 0.002) and IL-13 by 53% (P= 0.003), and also, blood flow by 18% (P= 0.0004). The control protocol had no effect on any of the variables assessed. CONCLUSIONS: Hypnosis reduced several components of the systemic and mucosal inflammatory response in active ulcerative colitis toward levels found previously in the inactive disease. Some of these effects may contribute to the anecdotally reported benefits of hypnotherapy and provide a rationale for controlled trials of hypnotherapy in UC. PMID- 18510608 TI - Minimal hepatic encephalopathy in patients with extrahepatic portal vein obstruction. AB - BACKGROUND AND AIMS: Minimal hepatic encephalopathy (MHE) is associated with poor quality of life and increased work disability in cirrhotic patients. Its prevalence in extrahepatic portal vein obstruction (EHPVO) is not known. We studied the prevalence of MHE in EHPVO patients and utility of critical flicker frequency (CFF) for diagnosing MHE. PATIENTS AND METHODS: Thirty-four EHPVO patients with a history of variceal bleed (age 23.2 +/- 11.2 yr, M:F 22:12) diagnosed by either Doppler US or MR angiography, which demonstrated portal vein obstruction and/or portal vein cavernoma, were evaluated by psychometry (number connection tests A, B or figure connection tests A, B) and P300 auditory event related potential (P300ERP). CFF was also evaluated. MHE was diagnosed by abnormal psychometry (>2 standard deviation [SD]) and/or P300ERP (>2.5 SD). RESULTS: Prevalence of MHE (N = 12) was 35.3%. Of 34 patients, P300ERP was abnormal (380.0 +/- 28.9 msec) in 11 (32%), psychometry in 9 (26.4%), both P300ERP and psychometry in 8 (23.5%), and CFF <38 Hz in 7 (21%) patients. Six (67%) patients with abnormal psychometry and 7 (64%) with abnormal P300ERP had CFF below 38 Hz. CFF had sensitivity (75%), specificity (96%), positive predictive value (86%), negative predictive value (93%), and diagnosis accuracy of 91% when compared to patients with both abnormal psychometry and P300ERP. The venous ammonia level was higher in patients with MHE (83.1 +/- 29.7 vs 44.7 +/- 16.1 micromol/L, P < 0.001) compared to patients without MHE. Spontaneous shunts were present in 67% of patients with MHE compared to 14% of non-MHE patients. MHE was more common in patients with spontaneous shunts (72.7%vs 17.4%, P= 0.001) than without spontaneous shunts. CONCLUSIONS: Prevalence of MHE in EHPVO patients is 35.3%, and CFF alone can reliably diagnose 88% of MHE patients with both abnormal psychometry and P300ERP. However, in view of the relatively low number of patients with MHE, the usefulness of CFF in this setting awaits confirmatory studies. PMID- 18510609 TI - Comparison of an established simple office-based immunological FOBT with fecal tumor pyruvate kinase type M2 (M2-PK) for colorectal cancer screening: prospective multicenter study. AB - OBJECTIVES: The immunological fecal occult blood test (IFOBT) has established itself as a more precise marker for colorectal cancer (CRC) screening than traditional guaiac-based FOBT. The simpler, cheaper, and more convenient newer office-based IFOBTs have been validated for diagnosing CRC. Dimeric isoenzyme of pyruvate kinase, M2-PK, expressed by tumor cells, has as well been proposed as a screening tool for CRC. This is the first study comparing fecal M2-PK as a screening biomarker for CRC against previously evaluated office-based IFOBT and colonoscopy. METHODS: Six hundred forty consecutive subjects (symptomatic, as well as for CRC screening) referred for colonoscopy for various indications across five centers in Germany provided the stool samples for performing M2-PK and an immunochemical FOB strip test. The IFOBT used was a rapid immunochromatographic assay for detection of fecal hemoglobin. For M2-PK, a commercially available sandwich enzyme-linked immunosorbent assay (ELISA) was used. The M2-PK test needs 6 h, while the office-based test can be read in just 10 min and is five times cheaper. RESULTS: Office-based IFOBT had sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and positive and negative likelihood ratios (LR) of 64.5, 96.3, 72.0, 94.9, 17.5, and 0.4 for diagnosing colorectal neoplasia (CRN), while the above performance characteristics for M2-PK at a cutoff value of 4 U/mL were 72.4, 73.8, 29.0, 94.8, 2.8, and 0.8 respectively. CONCLUSIONS: This office-based IFOBT was found to have significantly higher specificity, PPV, and positive LR as compared with M2-PK. IFOBT proved to be a convenient, noncumbersome, quick, and cheap tool in patients with above-average risk for detection of CRN. PMID- 18510610 TI - A pilot study of the association of low plasma adiponectin and Barrett's esophagus. AB - BACKGROUND AND AIMS: Gastroesophageal reflux disease (GERD) and obesity are associated with esophageal adenocarcinoma (EAC). We hypothesized that the obesity EAC relation is mediated by factors secreted from adipocytes. Adiponectin is a peptide secreted by adipocytes, and its plasma levels are inversely associated with obesity. We aimed to estimate the effect of circulating adiponectin on the risk of Barrett's esophagus (BE), an accepted precursor of EAC, controlling for GERD symptoms and other potential confounders. METHODS: We conducted a case control study in cases of BE compared with controls without BE; most controls had GERD. Odds ratios (OR), corresponding to associations with BE, were estimated from conditional and unconditional logistic regression analyses of 50 matched pairs. RESULTS: BE was inversely associated with plasma adiponectin level (OR for each 10-microg/mL decrement 4.7, 95% confidence interval [CI] 1.4-15.0) and positively associated with GERD duration >or=10 yr, male gender, tobacco smoking, body mass index (BMI), waist circumference, and waist-to-hip ratio. Further adjustment for GERD duration, tobacco use, and BMI increased the adiponectin-BE association (OR 6.4, 95% CI 1.1-37.0), but the estimated OR was reduced when adjusting for measures of abdominal obesity (e.g., OR 2.5, 95% CI 0.49-13.00) and further adjusting for gender (OR 1.8, 95% CI 0.66-4.70). CONCLUSIONS: Despite methodologic limitations, including the small sample size, our findings suggest that adiponectin may be involved in the etiology of BE. Rather than simply a mechanical effect of obesity promoting GERD, the effects of abdominal obesity on the risk of BE might be mediated by adiponectin and other circulating factors. PMID- 18510611 TI - Screening of 214 single nucleotide polymorphisms in 44 candidate cancer susceptibility genes: a case-control study on gastric and colorectal cancers in the Japanese population. AB - OBJECTIVES: Single nucleotide polymorphisms (SNPs) have potential as markers for identifying genes responsible for common diseases and for personalized medicine. To investigate the association between polymorphisms and gastrointestinal (gastric and colorectal) cancer, we performed a hospital-based case-control study in Japan. METHODS: We screened a total of 214 SNPs in 44 candidate genes by using a mass spectrometry-based technique (MassARRAY; Sequenom, Inc., San Diego, CA). In this study, 153 patients and 302 controls for gastric cancer and 121 patients and 245 controls for colorectal cancer were matched with regard to age, sex, and residential area. Genes were selected based on their possible interactions with the environment and lifestyle, and the candidate genes constitute those encoding xenobiotic metabolism enzymes, DNA repair enzymes, and other stress-related proteins. Each polymorphism was tested in controls to ensure its fit with Hardy Weinberg equilibrium. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by conditional logistic regression analysis to estimate the association between genetic polymorphisms and the risk of gastric and colorectal cancers. RESULTS: Twenty-one SNPs in nine genes were associated with the risk of gastric cancer (P < 0.05) and 15 SNPs in nine genes were associated with the risk of colorectal cancer (P < 0.05). CONCLUSIONS: The findings of the present study will be the basis for future large-scale association studies of gene-environment factors using the candidate gene approach for the Japanese population. PMID- 18510612 TI - Relation of bowel habits to fecal incontinence in women. AB - BACKGROUND: Though most women with fecal incontinence (FI) have anorectal dysfunctions, a majority have intermittent symptoms. Variations in bowel habits and daily routine may partly explain this. AIM: To compare bowel habits and daily routine between controls and FI, and between continent and incontinent stools among women with FI. METHOD: Using a mailed questionnaire, we identified 507 women with FI among 5,300 women in Olmsted County, MN. Bowel habits were compared among 127 randomly selected controls and 154 women with self-reported FI, who did ("active" FI, N = 106) or did not ("inactive" FI, N = 48) have an incontinent episode during a 2-wk bowel diary period. RESULTS: Independent risk factors for FI were: rectal urgency (odds ratio [OR] for inactive FI vs controls 5.6, 95% confidence interval [CI] 2.3-13.3; and OR for active FI vs inactive FI 2.0, 95% CI 0.9-4.3) and a sense of incomplete evacuation (OR for inactive FI vs controls 3.5, 95% CI 1.4-8.8; and OR for active FI vs inactive FI 2.2, 95% CI 1.1-4.9). Similar results were found for stool frequency and form. Among incontinent women, incontinent stools (versus continent stools) were less formed, more likely to occur at work, and to be preceded by rectal urgency. CONCLUSIONS: Bowel patterns, rectal urgency, and daily routine influence the occurrence of FI. Stool characteristics explained 46% of the likelihood for incontinence episodes, emphasizing that anorectal sensorimotor dysfunctions must also contribute to FI in women. PMID- 18510613 TI - Value of the apparent diffusion coefficient for quantification of low-grade hepatic encephalopathy. AB - BACKGROUND AND AIMS: Minimal hepatic encephalopathy (HE) is associated with poorer quality of life and increased work disability. Recently, low-grade cerebral edema has been implicated in chronic liver disease. METHODS: We measured the apparent diffusion coefficient (ADC) of water in various regions of the brains of patients with cirrhosis, and elucidated the significance of the evaluation of ADC in quantifying low-grade HE and predicting overt HE and survival. Forty patients with cirrhosis and 24 controls underwent diffusion weighted imaging, and patients were followed up every month. RESULTS: The mean ADC values were increased in cirrhotic patients with minimal HE versus no HE or controls. Minimal HE patients separated from no HE patients with a sensitivity of 70 approximately 90% and a specificity of 85 approximately 90%. ADC values correlated with individual neuropsychological tests. ADC values of white matter, such as the frontal (log-rank test 4.35, P < 0.05) and parietal (log-rank test 5.98, P < 0.05) white matter, was predictive of further bouts of overt HE. CONCLUSIONS: ADC is a reliable tool for quantification of low-grade HE, and could predict the development of overt HE. PMID- 18510614 TI - Biliary complications in donors for living donor liver transplantation. AB - OBJECTIVES: With the increasing number of living donor liver transplantations, biliary complications in donors have emerged as a major postoperative problem. The aim of the present study was to characterize the features of the biliary complications that occur in donors. METHODS: The study subjects comprised 731 consecutive patients who donated liver grafts (434 right-lobe and 297 left-lobe grafts) for transplantation at Kyoto University Hospital from July 1999 to December 2006. Donors whose biliary complications could not be cured by conservative therapy were referred for endoscopic treatment. RESULTS: Postoperative biliary complications occurred in 55 (7.5%) donors. Initially, 48 of these 55 donors had biliary leakage and 7 had biliary stricture. Subsequently, 5 of 48 donors with leakage developed biliary stricture. The respective incidences of biliary leakage and overall biliary complications were significantly higher among donors of right-lobe grafts (9.9% and 11.1%) than among donors of left-lobe grafts (1.7% and 2.4%). Among 55 donors with biliary complications, 24 were cured by conservative therapy, and 1 was converted to surgical repair due to ileus. Endoscopic treatment was successful in 24 of 30 (80%) donors treated by endoscopic retrograde cholangiography, while the remaining 6 (20%) patients underwent surgery due to difficulties with cannulation (N = 2), excessive biliary leakage (N = 2), or complete biliary obstruction (N = 2). CONCLUSIONS: Donors of right-lobe grafts have a significantly higher incidence of biliary complications than donors of left-lobe grafts. When conservative therapy fails, endoscopic treatment is effective for these complications, and should be attempted as the first-line therapy before surgical repair. PMID- 18510615 TI - In search of an ideal biomarker for nonalcoholic fatty liver disease. PMID- 18510616 TI - Improved survival in young women with colorectal cancer. AB - BACKGROUND: Studies have reported the effect of gender in the context of assessing predictors of survival from colorectal cancer (CRC); however, few have specifically addressed the impact of gender on the clinical and pathological outcomes of CRC. Appreciation of gender disparities may assist in the implementation of measures to address these differences, and improve the overall outcomes of patients with CRC. METHODS: The South Western Sydney Colorectal Tumour Group registry, which encompasses a population in excess of 800,000, prospectively collects data on new patients with CRC. Data from 1997 to 2004 were collected, including demography, site, grade, histopathology, stage, treatment, and survival. RESULTS: In total, 2,050 consecutive patients (44% women) with CRC were analyzed. Compared to men, women were older (median 69 yr, range 27-95 yr vs 67, range 22-92 yr, P= 0.001), had more emergency surgery for CRC-related complications (18.8%vs 15.1%, P= 0.03), had more proximal cancers (42.2%vs 31.5%, P < 0.001), had more poorly differentiated cancers (16.9%vs 12.9%, P= 0.01), and had fewer radiotherapy treatments for Dukes B and C rectal cancers (36.4%vs 48.1%, P= 0.02). Young women (aged 50 yr and below) had significantly better overall survival compared to young men; in this group, female gender predicted improved overall survival independent of age, emergency surgery, site, grade, and stage (hazard ratio [HR] 0.46, 95% confidence interval [CI] 0.25-0.86, P= 0.01). Similarly, young women had significantly better cancer-specific survival (HR 0.46, 95% CI 0.25-0.85, P= 0.01). However, older women (aged over 50 yr) had worse survival independent of age, emergency surgery, site, grade, and stage (HR 1.38, 95% CI 1.14-1.68, P= 0.001). There were no gender differences in screening, histopathology, stage, or utilization of chemotherapy. CONCLUSIONS: This study demonstrated an opposing effect of gender on overall and cancer-specific survival at either side of the age of 50 yr. The protective effect of estrogen on CRC may be an important factor. Women had a greater proportion of emergency surgery, which was related to the predominance of proximal cancers in this gender. Women also had more proximal cancers, thereby limiting flexible sigmoidoscopy as a screening test. PMID- 18510617 TI - Sporadic duodenal adenoma and the association with colorectal neoplasia: a case control study. AB - OBJECTIVES: Sporadic duodenal adenomas are an uncommon finding. It is not clear whether patients with sporadic duodenal adenoma have a greater risk for colorectal neoplasia and should undergo colonoscopy. The aims of the present study were to estimate the prevalence of colorectal neoplasia in patients with sporadic duodenal adenoma, and to compare colorectal neoplasia rates in patients with sporadic duodenal adenomas versus those without them. METHODS: A retrospective case-control study was conducted to identify sporadic duodenal adenoma patients using the databases of two academic and one regional hospital in the Netherlands. Colonoscopic findings in the sporadic duodenal adenoma patients were compared with those of a control group of patients who underwent both gastroduodenoscopy and colonoscopy. Furthermore, the frequency of colorectal cancer in the sporadic duodenal adenoma patients was compared with the population incidence of colorectal cancer. RESULTS: During the period 1991-2006, 102 patients in total with sporadic duodenal adenomas were identified. Colonoscopy was performed in 49 patients (48%), and colorectal neoplasia was present in 21 of these patients (43%). There was a significantly higher rate of both colorectal neoplasia (43%vs 17%, odds ratio [OR] 3.6, 95% confidence interval [CI] 1.7-7.4) and advanced colorectal adenoma (18%vs 3%, OR 7.8, 95% CI 2.1-29.4) in the patients with sporadic duodenal adenoma compared to that in the control group. Also, the incidence of colorectal cancer was higher in sporadic duodenal adenoma patients compared to that in the population (P= 0.02). CONCLUSIONS: Individuals with sporadic duodenal adenomas appear to be at a significantly higher risk of colorectal neoplasia, and therefore should undergo colonoscopy. PMID- 18510618 TI - Increased hepatic and circulating interleukin-6 levels in human nonalcoholic steatohepatitis. AB - BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) and type 2 diabetes (T2D) are growing public health problems, and are strongly associated. The link between the two conditions remains poorly understood. Hepatic interleukin-6 (IL-6), a major proinflammatory cytokine, expression is increased in animal models of NAFLD, while in mice, selective sustained upregulation of IL-6 in the liver results in systemic insulin resistance. The extent and clinical significance of hepatic IL-6 expression in human NAFLD, as well as potential mechanisms by which steatosis may increase IL-6 production in the liver, have not been examined. AIMS: To ascertain the occurrence and significance of IL-6 expression in the liver in human NAFLD. PATIENTS AND METHODS: Plasma was obtained at time of liver biopsy from 50 consecutive patients with suspected NAFLD. Histology was assessed blindly. Hepatic IL-6 expression was assessed by immunohistochemistry, while plasma IL-6 levels were determined by an enzyme-linked immunosorbent assay. RESULTS: IL-6 expression was markedly increased in the livers of patients with nonalcoholic steatohepatitis (NASH) as compared to patients with simple steatosis (P < 0.005) or normal biopsies (P < 0.010), confirming the presence of hepatic IL 6 expression in human NASH. A positive correlation was observed between hepatocyte IL-6 expression and degree of inflammation and stage of fibrosis. Furthermore, liver IL-6 expression positively correlated with plasma IL-6 levels and degree of systemic insulin resistance. Culture of liver cells with saturated, but not mono- or polyunsaturated, FFA resulted in a significant increase in IL-6 messenger RNA (mRNA) and protein expression. CONCLUSION: Collectively, these data suggest that increased hepatic IL-6 production may play an important role in NASH development, as well as in systemic insulin resistance and diabetes. PMID- 18510619 TI - Somatic pain sensitivity in children with recurrent abdominal pain. AB - OBJECTIVE: Evidence is accumulating that recurrent abdominal pain (RAP) in children is associated with visceral hyperalgesia. However, it is not known whether somatic sensitivity is altered as well. Therefore, the aim of our study was to assess somatic pain sensitivity in children with RAP and healthy controls at the abdomen and a distal site (thenar). METHODS: We examined 20 children with RAP (age 8-14) and 23 healthy control children (age 9-14). Heat and mechanical pain thresholds as well as measures of perceptual sensitization in response to repetitive mechanical or tonic thermal noxious stimulation were assessed. RESULTS: At the abdominal site, pain sensitivity in children with RAP did not differ significantly when compared to controls. At the thenar, pain thresholds of children in the RAP group were not significantly different from control children. However, children with RAP showed less perceptual sensitization in response to tonic heat and repetitive mechanical stimuli (ps or=56 mm was strongly suggestive of TMC (sensitivity 90%, specificity 90%, area under the ROC curve 0.91). No child with TMC died and 70% required colectomy during admission. Two of the three with intact colons at discharge required second-line therapy during the subsequent year. CONCLUSIONS: Colonic dilatation >or=56 mm in children with IBD strongly suggests TMC, if clinical signs are present. Mental alteration and hypotension may be less common in children than in adults. TMC in children with IBD is associated with poor outcome, with a high rate of corticosteroid failure. PMID- 18510625 TI - Colon cleansing with oral sodium phosphate in adolescents: dose, efficacy, acceptability, and safety. AB - BACKGROUND AND AIMS: Standardized bowel preparation in children and adolescents has not been established. Our aim was to compare two bowel preparation regimens and determine which was more effective, acceptable, and safer for children undergoing colonoscopy. METHODS: We compared the efficacy and acceptability of a 1-day regimen with oral sodium phosphate solution (NaP solution) (1 mL/kg/day, maximum 90 mL in two divided doses; regimen A) to our standard 3-day regimen magnesium citrate (4 mL/kg/day x 3 days, maximum 237 mL, followed by an enema the morning of colonoscopy; regimen B). After informed consent was obtained, 48 children were randomized (N = 25, 23, respectively). Weight, electrolytes, calcium, phosphorus, and magnesium were measured at screening and the day of the colonoscopy. Questionnaires were given to assess acceptability and adverse events. Endoscopists rated the quality of bowel preparation on a 4-level scale from excellent to poor. RESULTS: Median age and weight at screening were 14 yr, 53 kg, and 15 yr, 51 kg in regimen A and B, respectively. No statistical significance was observed in electrolytes, phosphorus, or adverse events apart from higher nausea intensity in regimen A (P= 0.012). Bowel cleansing was similar between groups (71% excellent or good). Subjects were more willing to repeat regimen A than B (77%vs 32%, respectively, P < 0.006). All 10 subjects who received regimen A and had prior colonoscopies using regimen B, preferred regimen A. CONCLUSIONS: In a selected group of otherwise healthy children and adolescents over 10.5 yr and above 34 kg, 1-day oral NaP solution was more acceptable than 3 day magnesium citrate with an enema, and both regimens were found to be safe and efficacious. PMID- 18510626 TI - Regulatory face-off: what agency should oversee face transplants? PMID- 18510627 TI - Prognostic value of cardiovascular screening in potential renal transplant recipients: a single-center prospective observational study. AB - We assessed the outcome of pretransplant cardiac assessment in a single center. Three hundred patients with end-stage renal disease underwent electrocardiogram, Bruce exercise testing (ETT) and ventricular assessment by cardiac MRI. Patients with high index of suspicion of coronary artery disease (CAD) underwent coronary angiography and percutaneous coronary intervention (PCI) if indicated. Two hundred and twenty-two patients were accepted onto the renal transplant waiting list; 80 patients were transplanted during the follow-up period and 60 died (7 following transplantation). Successful transplantation was associated with improved survival (mean survival 4.5 +/- 0.6 years vs. listed not transplanted 4.1 +/- 1.4 years vs. not listed 3.1 +/- 1.7 years; p < 0.001). Ninety-nine patients underwent coronary angiography; 65 had normal or low-grade CAD and 34 obstructive CAD. Seventeen patients (5.6%) were treated by PCI. There was no apparent survival difference between patients who underwent PCI or coronary artery bypass graft compared to those who underwent angiography without intervention or no angiography (p = 0.67). Factors associated with nonlisting for renal transplantation included burden of preexisting cardiovascular disease, poor exercise tolerance and severity of CAD. Pretransplant cardiovascular screening provides prognostic information and information that can be used to restrict access to transplantation. However, if the aim is to identify and treat CAD, the benefits are far from clear. PMID- 18510628 TI - PD-1 expression and IL-2 loss of cytomegalovirus- specific T cells correlates with viremia and reversible functional anergy. AB - Cytomegalovirus (CMV) represents a major cause of infectious complications after transplantation. Recently, chronic infections with lymphocyte choriomeningitis virus (LCMV), HIV or HCV were shown to be associated with functionally exhausted T cells characterized by high expression of the programmed death (PD)-1 molecule and altered cytokine expression patterns. We therefore hypothesized that functional exhaustion of CMV-specific CD4 T cells may determine impaired CMV control in patients after renal transplantation. In viremic transplant recipients, a significantly higher proportion of CMV-specific CD4 T cells was PD 1 positive (median 40.9%, 17.0-88.7%) as compared to nonviremic transplant patients (8.8%, 0.8-80.5%), dialysis patients (8.8%, 0-36.7%) or controls (3.2%, 0.3-15.4%, p < 0.0001). In line with functional impairment, PD-1-positive T cells produced significantly less IFNgamma as compared to PD-1-negative T cells (p < 0.0001). Moreover, unlike controls or nonviremic patients, CMV-specific T cells from viremic patients showed a significant loss of IL-2 production (p < 0.0001). Interestingly, functional anergy of CMV-specific CD4 T cells was reversible in that antibody-mediated blockade of PD-1 signaling with its ligands PD-L1/-L2 led to an up to 10-fold increase in CMV-specific proliferation. In conclusion, expression of PD-1 defines a reversible defect of CMV-specific CD4 T cells that is associated with viremia, and blocking PD-1 signaling may provide a potential target for enhancing the function of exhausted T cells in chronic CMV infection. PMID- 18510629 TI - Pulmonary hypertension in patients with bronchiolitis obliterans syndrome listed for retransplantation. AB - Bronchiolitis Obliterans Syndrome (BOS) is a major cause of morbidity and mortality post-lung transplantation. Pulmonary hypertension (PH) may complicate the course of patients with advanced lung disease. We sought to characterize the prevalence of PH in patients with BOS. We performed a retrospective analysis of lung transplant recipients with BOS relisted for transplantation with the United Network for Organ Sharing (UNOS). Right heart catheterization (RHC) data were required for analysis. Eighty patients with BOS qualified for the analysis. PH was present in 32.5% of patients with an average mean pulmonary artery pressure (mPAP) of 32.3 mmHg (range: 26-63 mmHg). Of these, 42.3% had an elevated pulmonary capillary wedge pressure. There was no difference in PH prevalence between bilateral (26.5%) and single lung recipients (41.9%), nor did it differ by primary disease. There was no correlation between pulmonary function data and the presence or severity of PH. There was no difference in oxygen requirements or 6-min walk distance between patients with and without PH. This is the first report of PH in patients with BOS. Many of these cases occur in association with diastolic dysfunction. Although no impact on functional status or outcomes was discerned, further studies appear warranted. PMID- 18510630 TI - Predictive value of dobutamine stress echocardiography for coronary artery disease detection in liver transplant candidates. AB - Patients with obstructive coronary artery disease (CAD) undergoing orthotopic liver transplantation (OLT) are at increased risk of poor outcomes. The accuracy of dobutamine stress echocardiography (DSE) to detect obstructive CAD is not well established in this population. We retrospectively identified patients with end stage liver disease who underwent both DSE and coronary angiography as part of risk stratification prior to OLT. One hundred and five patients had both DSE and angiography, of whom 14 had known CAD and 27 failed to reach target heart rate during DSE. Among the remaining 64 patients (45 men; average age 61 +/- 8 years) DSE had a low sensitivity (13%), high specificity (85%), low positive predictive value (PPV) (22%) and intermediate negative predictive value (NPV) (75%) for obstructive CAD. DSE as a screening test for obstructive CAD in OLT candidates has a poor sensitivity. The frequent chronotropic incompetence and low sensitivity in patients who achieve target heart rate, even in those with multiple cardiovascular disease risk factors, suggest that alternative or additional methods of risk stratification are necessary. PMID- 18510631 TI - Essential role of sphingosine-1-phosphate receptor 1-bearing CD8+CD44+CCR7+ T cells in acute skin allograft rejection. AB - A subset of naturally formed sphingosine-1-phosphate receptor 1 (S1P1)-bearing CD8(+)CD44(+)CCR7(+) memory T cells has been identified in transplant recipient BALB/c (H-2(d)) mice. The frequency of this subset of memory T cells is significantly increased in the spleen, lymph nodes and skin grafts in the recipient BALB/c mice during acute skin allograft rejections. The immune reconstitution with CD8(+)CD44(+)CCR7(+)S1P1(+) memory T cells facilitates acute skin allograft rejection in SCID mice. Being Th1-polarized and cytotoxic, CD8(+)CD44(+)CCR7(+)S1P1(+) memory T cells proliferate and differentiate immediately into effectors upon encountering allo-antigens. A siRNA against S1P1 inhibits CD8(+)CD44(+)CCR7(+)S1P1(+) memory T cell-mediated acute skin allograft rejection in SCID mice by means of knocking-down S1P1-expression. CCL21 mutant (CCL21-DeltaCT) has been used to compete with wild-type CCL21 in the course of binding to CCR7. Combined administration of siRNA S1P1 and CCL21-DeltaCT significantly prolongs the survival of skin allograft in the recipient BALB/c mice by means of inhibiting accumulation of CD8(+)CD44(+)CCR7(+)S1P1(+) memory T cells in the spleen and the skin grafts. Our data provide direct evidence that S1P1 and CCR7 are involved in the proliferation and trafficking of CD8(+)CD44(+)CCR7(+)S1P1(+) memory T cells. S1P1 may serve as a functional marker for CD8(+)CD44(+)CCR7(+) memory T cells. Targeting CD8(+)CD44(+)CCR7(+)S1P1(+) T cells may be a useful strategy to prolong the survival of allograft transplant. PMID- 18510632 TI - Alemtuzumab (Campath-1H) and tacrolimus monotherapy after renal transplantation: results of a prospective randomized trial. AB - The lymphocyte-depleting antibody alemtuzumab was evaluated in a prospective randomized multicenter trial in deceased donor kidney transplantation. The 65 patients in the study group received induction with alemtuzumab followed by delayed tacrolimus monotherapy, while the 66 patients in the control group were started on tacrolimus in combination with mycophenolate mofetil and steroids. Tacrolimus levels of 8-12 ng/mL for the first 6 months and 5-8 ng/mL thereafter were aimed for in both groups. At 12 months the biopsy-proven rejection rate was 20% in the study group and 32% in the control group (p = 0.09). Patient survival at 1 year was 98% for both groups. Graft survival was 96% for the study group versus 90% for the control group (p = 0.18). Graft function was identical in both groups. Adverse events were similar in both groups apart for more CMV infections in the study group. At the end of the first year 82% of the patients in the study group were steroid-free and 71% continued on tacrolimus monotherapy. These results suggest that alemtuzumab induction together with tacrolimus monotherapy is at least as efficient in renal transplantation as is a tacrolimus-based triple drug regimen with a similar safety profile but more CMV infections. PMID- 18510633 TI - Mammalian target of rapamycin inhibitor dyslipidemia in kidney transplant recipients. AB - The incidence, pathogenesis, consequences and treatment of mammalian target of rapamycin (mTOR) inhibitor dyslipidemia are not well described. We conducted a systematic review of randomized controlled trials reporting cholesterol and triglycerides in mTOR versus non-mTOR inhibitor immunosuppressive treatment regimens in kidney transplant recipients. All but one of 17 trials reported higher levels of cholesterol and triglycerides, or an increased prevalence of treatment with lipid-lowering agents. Approximately 60% of mTOR inhibitor-treated patients received lipid-lowering agents (2-fold higher than controls). There appeared to be little difference between dyslipidemias caused by sirolimus (14 trials) versus everolimus (3 trials). It was difficult to determine the extent to which declines in lipids over time posttransplant were due to lipid-lowering therapy, changes in doses and/or discontinuations of mTOR inhibitors. From the four trials that measured lipoproteins, it appeared that at least some of the increase in total cholesterol with mTOR inhibitors was due to increased low density lipoprotein cholesterol. What direct or indirect effects mTOR inhibitors have on atherosclerotic cardiovascular disease in kidney transplant patients are unknown. However, in the absence of the necessary clinical trials, dyslipidemia should be managed, as it would be in nontransplant patients at high risk for cardiovascular disease. PMID- 18510634 TI - Comparison of histidine-tryptophan-ketoglutarate and University of Wisconsin preservation in renal transplantation. PMID- 18510635 TI - Unexpected postmortem diagnosis of Acanthamoeba meningoencephalitis following allogeneic peripheral blood stem cell transplantation. AB - Meningoencephalitis caused by pathogenic free-living amebas is usually fatal. Only a few cases of Acanthamoeba meningoencephalitis, diagnosed at autopsy, have been reported following hematopoietic stem cell transplantation. We here report a case of Acanthamoeba meningoencephalitis following allogeneic peripheral blood stem cell transplantation with rapidly evolving neurologic symptoms that remained unexplained. Magnetic resonance imaging failed to show brain lesions and cerebrospinal fluid was negative for microbiological cultures. Definite diagnosis was an unexpected autopsy finding. As overall and teaching hospital autopsy rates are declining worldwide, we must emphasize the need of autopsy exams if we want to improve our knowledge as the best way to care for our patients. PMID- 18510636 TI - Cidofovir inhibits polyomavirus BK replication in human renal tubular cells downstream of viral early gene expression. AB - The human polyomavirus BK (BKV) causes nephropathy and hemorrhagic cystitis in kidney and bone marrow transplant patients, respectively. The anti-viral cidofovir (CDV) has been used in small case series but the effects on BKV replication are unclear, since polyomaviruses do not encode viral DNA polymerases. We investigated the effects of CDV on BKV(Dunlop) replication in primary human renal proximal tubule epithelial cells (RPTECs). CDV inhibited the generation of viral progeny in a dose-dependent manner yielding a 90% reduction at 40 microg/mL. Early steps such as receptor binding and entry seemed unaffected. Initial large T-antigen transcription and expression were also unaffected, but subsequent intra-cellular BKV DNA replication was reduced by >90%. Late viral mRNA and corresponding protein levels were also 90% reduced. In uninfected RPTECs, CDV 40 microg/mL reduced cellular DNA replication and metabolic activity by 7% and 11% in BrdU and WST-1 assays, respectively. BKV infection increased DNA replication to 142% and metabolic activity to 116%, respectively, which were reduced by CDV 40 microg/mL to levels of uninfected untreated RPTECs. Our results show that CDV inhibits BKV DNA replication downstream of large T-antigen expression and involves significant host cell toxicity. This should be considered in current treatment and drug development. PMID- 18510637 TI - FOXP3 expression in human kidney transplant biopsies is associated with rejection and time post transplant but not with favorable outcomes. AB - Expression of the transcription factor forkhead box P3 (FOXP3) in transplant biopsies is of interest due to its role in a population of regulatory T cells. We analyzed FOXP3 mRNA expression using RT-PCR in 83 renal transplant biopsies for cause in relationship to histopathology, clinical findings and expression of pathogenesis-based transcript sets assessed by microarrays. FOXP3 mRNA was higher in rejection (T-cell and antibody-mediated) than nonrejection. Surprisingly, some native kidney controls also expressed FOXP3 mRNA. Immunostaining for FOXP3 was consistent with RT-PCR, showing interstitial FOXP3+ lymphocytes, even in some native kidney controls. FOXP3 expression correlated with interstitial inflammation, tubulitis, interstitial fibrosis, tubular atrophy, C4d positivity, longer time posttransplant, younger donors, class II panel reactive antibody >20% and transcript sets reflecting inflammation and injury, but unlike these features was time dependent. In multivariate analysis, higher FOXP3 mRNA was independently associated with rejection, T-cell-associated transcripts, younger donor age and longer time posttransplant. FOXP3 expression did not correlate with favorable graft outcomes, even when the analysis was restricted to biopsies with rejection. Thus FOXP3 mRNA expression is a time-dependent feature of inflammatory infiltrates in renal tissue. We hypothesize that time-dependent entry of FOXP3 positive cells represents a mechanism for stabilizing inflammatory sites. PMID- 18510638 TI - Sirolimus may reduce fertility in male renal transplant recipients. AB - Assessment of sex hormones in organ transplant recipients suggests that sirolimus may impair testicular function. The aim of this study was to evaluate the frequency and severity of sirolimus-associated alterations in sperm parameters and their impact on fathered pregnancy rate. An observational study was carried out in male patients aged 20-40 years who received a kidney transplant during 1995-2005. Patients were sent a questionnaire by post, and sperm analysis was proposed. The fathered pregnancy rates according to the immunosuppressive regimen were estimated and compared using the Poisson model. Complete information was obtained from 95 out of 116 recipients. Patients treated with sirolimus throughout the post-transplant period had a significantly reduced total sperm count compared to patients who did not receive sirolimus (28.6 +/- 31.2 x 10(6) and 292.2 +/- 271.2 x 10(6), respectively; p = 0.006), and a decreased proportion of motile spermatozoa (22.2 +/- 12.3% and 41.0 +/- 14.5%, p = 0.01). Moreover, the fathered pregnancy rate (pregnancies/1000 patient years) was 5.9 (95% CI, 0.8 42.1) and 92.9 (95% CI, 66.4-130.0) in patients receiving sirolimus-based and sirolimus-free regimens, respectively (p = 0.007). Of six patients in whom sirolimus treatment was interrupted, only three showed a significant improvement in sperm parameters. Sirolimus is associated with impaired spermatogenesis and, as a corollary, may reduce male fertility. PMID- 18510639 TI - The association of state and national legislation with living kidney donation rates in the United States: a national study. AB - The effect of state legislation and federal policies supporting living donors on living kidney donation rates in the United States is unknown. We studied living kidney donation rates from 1988 to 2006, and we assessed changes in donation before and after the enactment of state legislation and the launch of federal initiatives supporting donors. During the study, 27 states enacted legislation. Among states enacting legislation, there was no statistically significant difference in the average rate of increase in overall living kidney donations after compared to before state legislation enactment (annual increase in donations per 1 000 000 population [95% confidence interval] 2.39 [1.94-2.84] compared to 1.68 [0.89-2.47] respectively, p > 0.05). Among states not enacting legislation, there was a statistically significantly greater annual increase in overall donation rates from 1997 to 2002 compared to before 1997 when federal initiatives commenced, but there was no growth in annual rates after 2002. State and federal legislation were associated with increases in living-unrelated donation. These findings suggest that although existing public policies were not associated with improvements in the majority of donations from living-related donors, they may have had a selective effect on barriers to living-unrelated kidney donation. PMID- 18510640 TI - Health status and renal function evaluation of kidney vendors: a report from Pakistan. AB - Unrelated kidney transplants have lead to commerce and kidney vending in Pakistan. This study on 104 vendors reports demographics, history, physical and systemic examination, ultrasound findings, renal and liver function and GFR by Cockcroft-Gault. Results were compared with 184 age, sex and nephrectomy duration matched living-related donors controls. Comparison of vendors versus controls showed mean age of 30.55 +/- 8.1 versus 30.65 +/- 7.85 (p = 0.91) years, M:F of 4.5:1 versus 4.2:1 and nephrectomy period of 33.89 +/- 30 versus 32.01 +/- 29.71 (p = 0.60) months respectively. Of the vendors 67% were bonded laborers earning <50 $/month as compared to controls where 68% were skilled laborers and self employed earning >100 $/month. History of vendors revealed jaundice in 8%, stone disease in 2% and urinary tract symptoms in 4.8%. Postnephrectomy findings between vendors versus donors showed BMI of 21.02 +/- 2.8 versus 23.02 +/- 4.2 (p = 0.0001), hypertension in 17% versus 9.2% (p = 0.04), serum creatinine (mg/dL) of 1.17+/-0.21 versus 1.02 +/- 0.27 (p = 0.0001), GFR (mL/min) of 70.94 +/- 14.2 versus 95.4 +/- 20.44 (p = 0.0001), urine protein/creatinine of 0.150 +/- 0.109 versus 0.10 +/- 0.10 (p = 0.0001), hepatitis C positivity in 27% versus 1.0% (p = 0.0001) and hepatitis B positive 5.7% versus 0.5% (p = 0.04), respectively. In conclusion, vendors had compromised renal function suggesting inferior selection and high risk for developing chronic kidney disease in long term. PMID- 18510641 TI - Baseline 6-min walk distance predicts survival in lung transplant candidates. AB - In a large, prospectively followed, two-center cohort of patients listed for lung transplantation (n = 376), we used Cox proportional hazards models to determine the importance of baseline 6-min walk distance (6MWD) in predicting patient survival. 6MWD used as a continuous variable was a significant predictor of survival after adjusting for other important covariates when transplant was considered as a time-varying covariate (HR for each 500 ft increase in 6MWD = 0.57, 95% CI: 0.43-0.77, p = 0.0002). 6MWD remained an important predictor of survival in models that considered only survival to transplant (HR for each 500 ft increase in 6MWD = 0.41, 95% CI: 0.27-0.62, p < 0.0001) or survival only after transplant (HR for each 500 ft increase in 6MWD = 0.40, 95% CI: 0.22-0.72, p = 0.002). Unadjusted Kaplan-Meier analysis demonstrates significantly different survival by 6MWD tertiles (<900, 900-1200, or >1200 ft, p-value = 0.0001). In the overall model, 6MWD prediction of survival was relatively homogeneous across disease category (6MWD by disease interaction term, p-value = 0.63). Our results demonstrate a significant relationship between baseline 6MWD and survival among patients listed for lung transplantation that exists across all native disease categories and extends through transplantation. The 6MWD is thus a useful measure of both urgency and utility among patients awaiting lung transplantation. PMID- 18510642 TI - Pharmacogenetics of immunosuppressants: progress, pitfalls and promises. AB - Most of the immunosuppressants used in organ transplantation are characterized by a narrow therapeutic index, whereby underdosing is associated with increased risk of rejection episodes and overdosing may exacerbate drug-related toxicity. Pharmacogenetics--complementary to pharmacokinetics--holds the potential to allow individualized dosing of immunosuppressive agents to optimize their therapeutic actions while minimizing adverse effects. Most of the studies have focused on polymorphisms of genes involved in drug metabolism and distribution, but as of now, only thiopurine-S-methyltransferase and cytochrome P 450 3A5 genotypes appear to have sufficiently large influence to have potentialities in guiding drug dosing. This may reflect the fact that available information from other polymorphisms derives almost exclusively from retrospective observations or from studies with important methodological biases. Active investigations aimed at identifying allelic variants of gene encoding for the pharmacologic targets are now ongoing. Recent studies have demonstrated that also donor genotype may play a significant role in immunosuppressive drug pharmacokinetics and pharmacodynamics. As one of the main future tasks, it is mandatory to develop mathematical models able to incorporate multiple gene polymorphisms with pharmacokinetic data and other critical information, providing algorithms able to individualize the best immunosuppressive therapy for each patient before transplantation. PMID- 18510643 TI - The spectrum of antibody-mediated renal allograft injury: implications for treatment. AB - Improvements in anti-HLA antibody detection and diagnostic criteria have increased recognition of antibody-mediated rejection (AMR) following renal transplantation. Therapy of acute AMR is directed toward rapidly lowering circulating donor-specific antibody (DSA) activity. Despite reversal of acute renal dysfunction, however, antibody-secreting plasma cells in spleen and bone marrow are not depleted by treatment and circulating DSA commonly remains detectable in peripheral blood. Sequential ultrastructural studies of renal allografts during acute AMR show progression of microvascular endothelial abnormalities from necrosis and apoptosis to glomerular and peritubular capillary basement membrane duplication, termed transplant glomerulopathy (TG), a manifestation of chronic AMR. Additionally, long-term exposure to anti-HLA antibodies (particularly against class II antigens) is associated with shortened allograft survival and TG even in the absence of documented acute AMR. The association of TG with prior acute AMR and with circulating DSA provides evidence that antibody-mediated allograft injury exists as a spectrum of renal injury. Although effective therapy is available for acute AMR, allografts remain at risk for chronic AMR and shortened survival. The optimum approach to treatment for chronic AMR remains to be determined. PMID- 18510644 TI - Rescue of a living donor with liver transplantation. AB - Postoperative liver failure is a rare complication after living donor liver resection. This is a case report of a 22-year-old healthy donor who was rescued with liver transplantation 11 days after right hemihepatectomy. Nine months later the patient is alive, and has fully recovered from his multiple organ failure. According to a review of the literature, there are four additional living liver donors, who received a liver transplant. Our own patient is the only survivor, so far. This case demonstrates that even in supposedly healthy living donors postoperative complications cannot be completely prevented. Although liver failure is rare in these patients, timely transplantation may need to be considered as the only life-saving treatment. PMID- 18510645 TI - Calcineurin inhibitor withdrawal after renal transplantation with alemtuzumab: clinical outcomes and effect on T-regulatory cells. AB - To address the results of calcineurin inhibitor (CNI) withdrawal after alemtuzumab induction relative to CNI continuation, we performed a pilot randomized clinical trial in renal allograft recipients on CNI, a mycophenolic acid derivative and steroids after the first 2 months posttransplantation. Forty patients were randomized to taper off CNI or to maintain it, and followed for at least 1 year. Four patients in the withdrawal group were treated for acute rejection while no patient received antirejection treatment in the control group. Two control patients withdrew CNI due to nephrotoxicity. Estimated GFR was similar in both groups after 1 year. Flow cytometry of CD4(+)CD25(+)CTLA 4(+)FoxP3(+) regulatory T cells (Treg) demonstrated a significant increase in Treg percentages in the peripheral blood of alemtuzumab-treated patients on CNI early postransplant. Furthermore, the increased Treg percentages in the withdrawal cohort were unchanged at month 6 postenrollment, whereas they decreased significantly in those patients maintained on CNI. Patients withdrawn from CNI after alemtuzumab trend toward a higher rejection rate, but most patients can be weaned from a CNI using this regimen. With the exception of maintaining increased Treg levels, the benefits are not appreciable in this short follow-up, and a larger randomized trial is justified. PMID- 18510647 TI - Linkage validation of RP25 Using the 10K genechip array and further refinement of the locus by new linked families. AB - Retinitis pigmentosa (RP) is a clinically and genetically heterogeneous group of retinal dystrophies, characterised by rod photoreceptor cell degeneration with autosomal recessive RP (arRP) as the commonest form worldwide. To date, a total of 26 loci have been reported for arRP, each having a prevalence of 1-5%, except for the RP25 locus which was identified as the genetic cause of 14% of arRP cases in Spain. In order to validate the original linkage of RP25, we undertook a total genome scan using the 10K GeneChip mapping array on three of the previously linked families. The data obtained supported the initial findings of linkage. Additionally, linkage analysis in 18 newly ascertained arRP families was performed using microsatellite markers spanning the chromosome 6p12.1-q15 interval. Five out of the 18 families showed suggestive evidence of linkage to RP25, hence supporting the high prevalence of this locus in the Spanish population. Furthermore, the finding of a crossover in one of these families is likely to have refined the disease interval from the original 16 cM to only a 2.67 cM region between D6S257 and D6S1557. PMID- 18510646 TI - Large-scale molecular analysis of a 34 Mb interval on chromosome 6q: major refinement of the RP25 interval. AB - A large scale bioinformatics and molecular analysis of a 34 Mb interval on chromosome 6q12 was undertaken as part of our ongoing study to identify the gene responsible for an autosomal recessive retinitis pigmentosa (arRP) locus, RP25. Extensive bioinformatics analysis indicated in excess of 110 genes within the region and we also noted unfinished sequence on chromosome 6q in the Human Genome Database, between 58 and 61.2 Mb. Forty three genes within the RP25 interval were considered as good candidates for mutation screening. Direct sequence analysis of the selected genes in 7 Spanish families with arRP revealed a total of 244 sequence variants, of which 67 were novel but none were pathogenic. This, together with previous reports, excludes 60 genes within the interval ( approximately 55%) as disease causing for RP. To investigate if copy number variation (CNV) exists within RP25, a comparative genomic hybridization (CGH) analysis was performed on a consanguineous family. A clone from the tiling path array, chr6tp-19C7, spanning approximately 100-Kb was found to be deleted in all affected members of the family, leading to a major refinement of the interval. This will eventually have a significant impact on cloning of the RP25 gene. PMID- 18510648 TI - Is post-mortem harm possible? Understanding death harm and grief. AB - The purpose of this article is not to affirm or deny particular philosophical positions, but to explore the limits of intelligibility about what post-mortem harm means, especially in the light of improper post-mortem procedures at Bristol and Alder Hey hospitals in the late 1990s. The parental claims of post-mortem harm to dead children at Alder Hey Hospital are reviewed from five different philosophical perspectives, eventually settling on a crucial difference of perspective about how we understand harm to the dead. On the one hand there is the broadly 'analytical' tradition(1) of thinking that predicates the notion of harm on the basis of an existing subject. Since the dead are non-existent persons, it makes little sense to view the dead as being harmed. On the other hand, there is a phenomenological perspective, where the dead, in respect to the experience of grief, are existentially absent. This forms the basis that it is possible to harm grieving parent's experiences of how their dead are treated. The article ends with a short examination of what harming the dead implies for traditional bioethical concerns, namely, obtaining informed consent from significant others when planning medical research on the newly dead. PMID- 18510649 TI - Testing marginal homogeneity against stochastic order in multivariate ordinal data. AB - SUMMARY: Many assessment instruments used in the evaluation of toxicity, safety, pain, or disease progression consider multiple ordinal endpoints to fully capture the presence and severity of treatment effects. Contingency tables underlying these correlated responses are often sparse and imbalanced, rendering asymptotic results unreliable or model fitting prohibitively complex without overly simplistic assumptions on the marginal and joint distribution. Instead of a modeling approach, we look at stochastic order and marginal inhomogeneity as an expression or manifestation of a treatment effect under much weaker assumptions. Often, endpoints are grouped together into physiological domains or by the body function they describe. We derive tests based on these subgroups, which might supplement or replace the individual endpoint analysis because they are more powerful. The permutation or bootstrap distribution is used throughout to obtain global, subgroup, and individual significance levels as they naturally incorporate the correlation among endpoints. We provide a theorem that establishes a connection between marginal homogeneity and the stronger exchangeability assumption under the permutation approach. Multiplicity adjustments for the individual endpoints are obtained via stepdown procedures, while subgroup significance levels are adjusted via the full closed testing procedure. The proposed methodology is illustrated using a collection of 25 correlated ordinal endpoints, grouped into six domains, to evaluate toxicity of a chemical compound. PMID- 18510650 TI - A comparison of methods for estimating the causal effect of a treatment in randomized clinical trials subject to noncompliance. AB - SUMMARY: We consider the analysis of clinical trials that involve randomization to an active treatment (T = 1) or a control treatment (T = 0), when the active treatment is subject to all-or-nothing compliance. We compare three approaches to estimating treatment efficacy in this situation: as-treated analysis, per protocol analysis, and instrumental variable (IV) estimation, where the treatment effect is estimated using the randomization indicator as an IV. Both model- and method-of-moment based IV estimators are considered. The assumptions underlying these estimators are assessed, standard errors and mean squared errors of the estimates are compared, and design implications of the three methods are examined. Extensions of the methods to include observed covariates are then discussed, emphasizing the role of compliance propensity methods and the contrasting role of covariates in these extensions. Methods are illustrated on data from the Women Take Pride study, an assessment of behavioral treatments for women with heart disease. PMID- 18510651 TI - On comparison of mixture models for closed population capture-recapture studies. AB - SUMMARY: A mixture model is a natural choice to deal with individual heterogeneity in capture-recapture studies. Pledger (2000, Biometrics 56, 434 442; 2005, Biometrics 61, 868-876) advertised the use of the two-point mixture model. Dorazio and Royle (2003, Biometrics 59, 351-364; 2005, Biometrics 61, 874 876) suggested that the beta-binomial model has advantages. The controversy is related to the nonidentifiability of the population size (Link, 2003, Biometrics 59, 1123-1130) and certain boundary problems. The total bias is decomposed into an intrinsic bias, an approximation bias, and an estimation bias. We propose to assess the approximation bias, the estimation bias, and the variance, with the intrinsic bias excluded when comparing different estimators. The boundary problems in both models and their impacts are investigated. Real epidemiological and ecological examples are analyzed. PMID- 18510652 TI - Simultaneous factor selection and collapsing levels in ANOVA. AB - When performing an analysis of variance, the investigator often has two main goals: to determine which of the factors have a significant effect on the response, and to detect differences among the levels of the significant factors. Level comparisons are done via a post-hoc analysis based on pairwise differences. This article proposes a novel constrained regression approach to simultaneously accomplish both goals via shrinkage within a single automated procedure. The form of this shrinkage has the ability to collapse levels within a factor by setting their effects to be equal, while also achieving factor selection by zeroing out entire factors. Using this approach also leads to the identification of a structure within each factor, as levels can be automatically collapsed to form groups. In contrast to the traditional pairwise comparison methods, these groups are necessarily nonoverlapping so that the results are interpretable in terms of distinct subsets of levels. The proposed procedure is shown to have the oracle property in that asymptotically it performs as well as if the exact structure were known beforehand. A simulation and real data examples show the strong performance of the method. PMID- 18510653 TI - Joint regression analysis of correlated data using Gaussian copulas. AB - This article concerns a new joint modeling approach for correlated data analysis. Utilizing Gaussian copulas, we present a unified and flexible machinery to integrate separate one-dimensional generalized linear models (GLMs) into a joint regression analysis of continuous, discrete, and mixed correlated outcomes. This essentially leads to a multivariate analogue of the univariate GLM theory and hence an efficiency gain in the estimation of regression coefficients. The availability of joint probability models enables us to develop a full maximum likelihood inference. Numerical illustrations are focused on regression models for discrete correlated data, including multidimensional logistic regression models and a joint model for mixed normal and binary outcomes. In the simulation studies, the proposed copula-based joint model is compared to the popular generalized estimating equations, which is a moment-based estimating equation method to join univariate GLMs. Two real-world data examples are used in the illustration. PMID- 18510654 TI - Power and sample size estimation for the Wilcoxon rank sum test with application to comparisons of C statistics from alternative prediction models. AB - The Wilcoxon Mann-Whitney (WMW) U test is commonly used in nonparametric two group comparisons when the normality of the underlying distribution is questionable. There has been some previous work on estimating power based on this procedure (Lehmann, 1998, Nonparametrics). In this article, we present an approach for estimating type II error, which is applicable to any continuous distribution, and also extend the approach to handle grouped continuous data allowing for ties. We apply these results to obtaining standard errors of the area under the receiver operating characteristic curve (AUROC) for risk prediction rules under H(1) and for comparing AUROC between competing risk prediction rules applied to the same data set. These results are based on SAS callable functions to evaluate the bivariate normal integral and are thus easily implemented with standard software. PMID- 18510655 TI - Bayesian nonparametric estimation of continuous monotone functions with applications to dose-response analysis. AB - In this article, we consider monotone nonparametric regression in a Bayesian framework. The monotone function is modeled as a mixture of shifted and scaled parametric probability distribution functions, and a general random probability measure is assumed as the prior for the mixing distribution. We investigate the choice of the underlying parametric distribution function and find that the two sided power distribution function is well suited both from a computational and mathematical point of view. The model is motivated by traditional nonlinear models for dose-response analysis, and provides possibilities to elicitate informative prior distributions on different aspects of the curve. The method is compared with other recent approaches to monotone nonparametric regression in a simulation study and is illustrated on a data set from dose-response analysis. PMID- 18510656 TI - Differential equation modeling of HIV viral fitness experiments: model identification, model selection, and multimodel inference. AB - Many biological processes and systems can be described by a set of differential equation (DE) models. However, literature in statistical inference for DE models is very sparse. We propose statistical estimation, model selection, and multimodel averaging methods for HIV viral fitness experiments in vitro that can be described by a set of nonlinear ordinary differential equations (ODE). The parameter identifiability of the ODE models is also addressed. We apply the proposed methods and techniques to experimental data of viral fitness for HIV-1 mutant 103N. We expect that the proposed modeling and inference approaches for the DE models can be widely used for a variety of biomedical studies. PMID- 18510657 TI - Renal surgery in the elderly: morbidity in patients aged >75 years in a contemporary series. AB - OBJECTIVES: To evaluate the surgical complications in a contemporary group of elderly patients with renal masses, as almost a quarter of patients with newly diagnosed renal mass are aged >74 years, with the potential for significant comorbidity. PATIENTS AND METHODS: From April 2004 to June 2007, of 379 surgical resections of renal tumours, we assessed 117 consecutive patients aged >or=75 years, who had either radical nephrectomy (RN) or partial nephrectomy (PN) for assumed renal cell carcinoma. Also elderly patients who had nephroureterectomy (NU) for upper urothelial cancer were followed. RESULTS: Fifty patients had RN, 57 PN and 10 had NU; the median (range) age of all patients was 78.1 (72.7-92.5) years and was similar in all groups. No patient died during surgery and only one died within 90 days. The complication rates during and after surgery RN, PN and NU were 12%, 15% and 20%, respectively; the major complications within 30 days were 4%, 7% and 10%; major complications included bleeding during surgery and one acute bleeding event after surgery in the PN group. CONCLUSIONS: Open renal surgery in elderly patients can be done safely; there was no difference in morbidity among RN, PN and NU. Renal surgery in the elderly patient is safe if done at a specialized centre. Mortality and morbidity can be very low, rendering this a feasible approach in the treatment of renal masses even if the prognosis is not determined by the oncological situation but by comorbidity. PMID- 18510658 TI - Outcome of dorsal buccal graft urethroplasty for recurrent bulbar urethral strictures. AB - OBJECTIVE: To audit our results of dorsal buccal mucosal graft urethroplasty for recurrent bulbar urethral stricture disease and compare them with those from specialist centres. PATIENTS AND METHODS: Data were collected prospectively on 52 men who had urethroplasty with > or =1 year of follow-up; failure was defined as the need for further intervention. RESULTS: The mean (range) age of the patients was 39 (19-61) years and 23 (45%) had an identifiable cause for their stricture. The mean (range) stricture length was 3.5 (1.5-6) cm and was associated with moderate or severe spongiofibrosis in 38 (73%) men. Ten (19%) men had minor complications after surgery. The mean (range) follow-up was 34 (12-80) months, with the mean maximum urinary flow rate increasing from 6 to 24 mL/s after surgery. The surgery failed, requiring dilatation or urethrotomy, in seven (14%) men at a mean (range) of 25 (15-50) months after urethroplasty, giving an overall success rate of 86%. CONCLUSION: This prospective audit of dorsal buccal patch augmentation urethroplasty for bulbar strictures shows an equivalent outcome to the standard set by the expert originators, suggesting that is transferable to less specialized centres. The efficacy, low complication rate, short hospital stay and general applicability of the technique encourage its use for all men with recurrent bulbar stricture disease, but formal comparison with other options in randomized trials, including cost-effectiveness analysis, is needed. PMID- 18510659 TI - Extended-release tolterodine with or without tamsulosin in men with lower urinary tract symptoms and overactive bladder: effects on urinary symptoms assessed by the International Prostate Symptom Score. AB - OBJECTIVE: To evaluate the efficacy of tolterodine extended-release (ER) plus tamsulosin on lower urinary tract symptoms (LUTS) as assessed by changes in the International Prostate Symptom Score (IPSS) in men who met symptom entry criteria for both overactive bladder (OAB) and benign prostatic hyperplasia (BPH) trials. PATIENTS AND METHODS: Men aged > or =40 years with an IPSS of > or =12 and diary documented OAB symptoms (> or =8 voids/24 h and > or =3 urgency episodes/24 h, with or without urgency urinary incontinence) who reported at least moderate problems related to their bladder condition were randomized to receive placebo, tolterodine ER (4 mg), tamsulosin (0.4 mg), or tolterodine ER (4 mg) + tamsulosin (0.4 mg) once daily for 12 weeks. Patients completed the IPSS at baseline and at 1, 6 and 12 weeks. RESULTS: Patients receiving tolterodine ER + tamsulosin had significantly greater improvements than those taking placebo on IPSS storage subscale scores and scores for all three individual storage items included on the IPSS (urinary frequency, urgency, and nocturnal micturitions) by 12 weeks. Storage subscale and urgency scores were significantly improved vs placebo at 1 and 6 weeks, whereas frequency scores were significantly improved at 6 weeks. Changes in IPSS storage subscale and individual storage item scores in the tolterodine ER and tamsulosin monotherapy groups were not significantly different from placebo at most time points. IPSS voiding subscale scores and scores for three of four individual voiding items (sensation of incomplete emptying, intermittency, and weak stream) were significantly improved by 12 weeks for patients receiving tamsulosin monotherapy vs placebo. Voiding subscale and intermittency scores were significantly improved vs placebo at 1 week; weak stream scores were significantly improved at 1 and 6 weeks. The IPSS voiding subscale and individual voiding item scores in the tolterodine ER + tamsulosin and tolterodine ER groups were not significantly different from placebo at most time points. CONCLUSIONS: In this distinct clinical research population of men who met traditional symptom entry criteria for both OAB and BPH trials, tolterodine ER + tamsulosin was significantly more effective than placebo in treating storage LUTS, including OAB symptoms. Tamsulosin monotherapy produced significant improvements in voiding LUTS. PMID- 18510660 TI - Extracorporeal shock-wave therapy for treating chronic pelvic pain syndrome: a feasibility study and the first clinical results. AB - OBJECTIVE: To investigate the feasibility and clinical outcome of extracorporeal shock-wave therapy (ESWT) for patients suffering from chronic pelvic pain syndrome (CPPS). PATIENTS AND METHODS: The study included 34 patients who had had CPPS for >or=3 months, who were investigated in two subsequent studies. ESWT was administered using a perineal approach with two different standard ESWT devices with and without an ultrasonographic positioning system. The follow-up was at 1, 4 and 12 weeks after ESWT, to evaluate the effects on pain, quality of life and voiding. Imaging studies and changes in prostate-specific antigen (PSA) were used to investigate the safety and side-effects of ESWT. RESULTS: All patients completed the treatments and follow-up; there were statistically significant improvements in pain and quality of life after ESWT. Voiding conditions were temporarily improved but with no statistical significance. Perineal ESWT was easy and safe to administer with no anaesthesia on an outpatient basis. Side-effects could be excluded clinically, by imaging studies and by changes in PSA level. CONCLUSION: Perineal ESWT must be considered as a promising new therapy for CPPS, in particular as it is easy to apply and causes no side-effects. PMID- 18510661 TI - The Canadian Uro-Oncology Group multicentre phase II study of docetaxel administered every 3 weeks with prednisone in men with metastatic hormone refractory prostate cancer progressing after mitoxantrone/prednisone. AB - OBJECTIVE: To investigate the use of docetaxel 75 mg/m(2) intravenously every 3 weeks plus prednisone 5 mg orally twice daily in men with metastatic hormone refractory prostate cancer (HRPC) progressing after first-line mitoxantrone/prednisone (MP), the primary outcome being progression-free survival with prostatic-specific antigen (PSA) and pain response, toxicity and quality of life (QoL) also assessed. PATIENTS AND METHODS: Thirty patients from four centres were enrolled in the study; all had had previous MP for symptomatic, metastatic HRPC and all had castrate levels of testosterone maintained during therapy. RESULTS: At enrolment, the median age was 69 years, the mean PSA level was 324 ng/dL, and 86% of patients reported pain. There was a PSA response in 57% of the men and a reduction in pain in >60%; the overall QoL was maintained. There were four cases of febrile neutropenia and two treatment-related deaths. The median progression-free and overall survival were 5 and 15 months, respectively. CONCLUSION: Docetaxel was associated with high rates of PSA and pain response in this study. Non-haematological toxicity was similar to that during first-line treatment, but rates of febrile neutropenia and toxic death appeared to be slightly higher. In selected patients with progressive metastatic HRPC previously treated with mitoxantrone, docetaxel appears to be a beneficial therapeutic option. PMID- 18510662 TI - Complementary and alternative medicine in urology: what we need to know in 2008. PMID- 18510663 TI - Dermoscopic monitoring of melanocytic skin lesions: clinical outcome and patient compliance vary according to follow-up protocols. AB - BACKGROUND: Dermoscopic monitoring of melanocytic lesions increases the likelihood that featureless melanomas are not overlooked and minimizes the excision of benign lesions. Objective To examine clinical outcome and patient compliance using different follow-up protocols. METHODS: A retrospective analysis of 600 lesions from 405 patients (aged 6-79 years) was performed to examine patient compliance and clinical outcome in patients with multiple atypical melanocytic lesions undergoing sequential dermoscopy imaging during short-, medium- or long-term follow-up. Based on the degree of dermoscopic atypical features, patients were scheduled for short-term monitoring with follow-up after 3 months, medium-term monitoring with follow-up after 6 months or long-term monitoring with annual follow-up. Criteria leading to excision of monitored lesions differed according to the follow-up protocol. RESULTS: In a median follow up period of 23 months, 54 (9%) lesions were excised, revealing 12 early melanomas (occurring in 3% of monitored patients), one basal cell carcinoma and 41 melanocytic naevi. The melanoma/benign ratio of excised lesions was 1 : 3.4. Seven of 12 melanomas showed changes after two to four visits, corresponding to 8 54 months of follow-up. Patient compliance was 84% for short-term monitoring, 63% for medium-term monitoring and 30% for long-term monitoring. CONCLUSIONS: In patients with multiple naevi sequential dermoscopy imaging is a useful strategy to avoid missing melanomas while minimizing unnecessary excision of benign lesions. For better compliance, the first re-examination should be scheduled at 3 months after the baseline visit. Regular annual follow-up monitoring is also needed to detect slow-growing melanomas in which subtle changes may become apparent only over time. PMID- 18510664 TI - para-Phenylenediamine: the profile of an important allergen. Results of the IVDK. AB - BACKGROUND: para-Phenylenediamine (PPD) is an important contact allergen and primarily used in hair dyeing. OBJECTIVES: To quantify cases of contact allergy (CA) to PPD attributed to sources of exposure. METHODS: Patients with PPD CA, diagnosed by the Information Network of Departments of Dermatology (IVDK; n = 3307 of 83 030 patients tested), were divided into five subgroups of exposure. Demographic variables and pattern of concomitant reactions were used to characterize the subgroups further. The impact of individual factors on the risk of PPD CA was examined with a logistic regression analysis. Clinical epidemiology and drug utilization research (CE-DUR) methods were employed to estimate the 10 year prevalence of PPD CA in the general population. RESULTS: The 4% prevalence of PPD CA in patients was extrapolated to a prevalence of 0.96% in the general population. The defined profiles were found to 'explain' the following percentages of PPD CA: (i) hair dyeing in clients 22% (0.2% of the general population); (ii) different occupational exposures, namely hair dyeing by hairdressers, paint- and rubber-associated exposures 23% (0.22% of the general population); (iii) clothing/shoes 12% (0.12% of the general population). A probable causal exposure to PPD could not be identified in about 44% of patients with PPD CA. CONCLUSIONS: In more than 50% of cases of PPD CA, a (very) probable causal exposure was identified. In the large remainder (44%) this was not possible. 'Historical' CA without current relevance and active sensitization through patch testing (a further important exposure to PPD) must be considered in these patients. PMID- 18510665 TI - An evidence base for reconsidering current follow-up guidelines for patients with cutaneous melanoma less than 0.5 mm thick at diagnosis. AB - BACKGROUND: Despite current guidelines, there is uncertainty about the required duration and frequency of follow-up visits for patients with invasive primary cutaneous melanoma < 0.5 mm thick. OBJECTIVES: To review patients with invasive melanoma thinner than 0.5 mm followed for at least 5 years to provide an evidence base for considering modification of guidelines. METHODS: A retrospective review of 430 patients diagnosed in the west of Scotland during 1992-2001 with melanoma < 0.5 mm was carried out. Recurrence, deaths from melanoma and second primary melanomas were all identified. RESULTS: From 1992 to 2001, 430 melanomas < 0.5 mm thick at diagnosis were diagnosed out of a total of 3036 primary cutaneous melanomas. To date there have been 593 deaths from melanoma (19%) in the whole group. Five of these deaths were reported in patients with melanomas < 0.5 mm, but on pathological review two were thicker than 0.5 mm at diagnosis (1.5 and > 3 mm), and the remaining three patients all developed thicker second primary melanomas (2.7, 12.0 and 19.0 mm) with a recurrence pattern and timing indicating that these thicker primaries were the cause of death. Fourteen further patients developed a second primary melanoma, and 13 are currently alive and disease free, one dying of other causes. CONCLUSIONS: Our data indicate that recurrence and subsequent death from melanomas < 0.5 mm is a very rare event, and that quarterly follow-up for 3 years will yield very few events. Modification of current guideline recommendations are suggested to include a period of patient education concentrating on recognition of second primary tumours followed by rapid access to an expert opinion if required. PMID- 18510666 TI - Glycerol and the skin: holistic approach to its origin and functions. AB - Glycerol is a trihydroxy alcohol that has been included for many years in topical dermatological preparations. In addition, endogenous glycerol plays a role in skin hydration, cutaneous elasticity and epidermal barrier repair. The aquaporin 3 transport channel and lipid metabolism in the pilosebaceous unit have been evidenced as potential pathways for endogenous delivery of glycerol and for its metabolism in the skin. Multiple effects of glycerol on the skin have been reported. The diverse actions of the polyol glycerol on the epidermis include improvement of stratum corneum hydration, skin barrier function and skin mechanical properties, inhibition of the stratum corneum lipid phase transition, protection against irritating stimuli, enhancement of desmosomal degradation, and acceleration of wound-healing processes. Even an antimicrobial effect has been demonstrated. Topical application of glycerol-containing products improves skin properties in diseases characterized by xerosis and impaired epidermal barrier function, such as atopic dermatitis. The increase of epidermal hydration by glycerol is critical in skin conditions aggravated by dry and cold environmental conditions, e.g. winter xerosis. This paper provides a review on effects of glycerol on the skin, the mechanisms of its action, and the potential applications of glycerol in dermatology. PMID- 18510667 TI - Ultraviolet responses of Gorlin syndrome primary skin cells. AB - BACKGROUND: Gorlin syndrome, or naevoid basal cell carcinoma syndrome (NBCCS), is an autosomal dominant disorder associated with mutations in the PTCH1 gene, which encodes the receptor of SONIC HEDGEHOG. In addition to developmental abnormalities, patients with NBCCS are prone to basal cell carcinoma (BCC), the most frequent type of nonmelanoma skin cancer in humans. OBJECTIVES: As ultraviolet (UV) exposure plays a prominent role in the development of sporadic BCC, we aimed to determine whether primary NBCCS skin cells exhibit differential responses to UV exposure compared with wild-type (WT) skin cells. METHODS: Primary fibroblast and keratinocyte strains were isolated from nonlesional skin biopsies of 10 patients with characteristic NBCCS traits. After identification of PTCH1 mutations, capacities of NBCCS cells to repair UV-induced DNA lesions and to survive after UV irradiation, as well as p53 responses, were compared with those of WT skin cells. RESULTS: The c1763insG PTCH1 mutation is described for the first time. DNA repair and cell survival analyses following UV irradiation revealed no obvious differences between responses of NBCCS and WT fibroblasts and keratinocytes. However, p53 accumulation after UV irradiation was abnormally persistent in all NBCCS primary keratinocyte strains compared with WT keratinocytes. CONCLUSIONS: Our observations that NBCCS cells harbour normal DNA repair and survival capacities following UV irradiation better explain that BCC proneness of patients with NBCCS does not solely concern body areas exposed to sunlight and suggest rather that it might be due to cell cycle alterations. PMID- 18510668 TI - Susceptibility of dermatophyte isolates obtained from a large worldwide terbinafine tinea capitis clinical trial. AB - BACKGROUND: Our group, in collaboration with seven other laboratories, has recently developed a method to determine the susceptibility of dermatophytes. OBJECTIVES: The objective of this study was to determine the terbinafine susceptibility profile of dermatophyte isolates obtained from patients with tinea capitis enrolled in two large worldwide clinical trials and to investigate whether these susceptibilities differ by geographical location. Methods Susceptibilities were determined according to the Clinical and Laboratory Standards Institute M38-A2 standard. RESULTS: From a total of 978 baseline dermatophyte isolates, we selected 301 isolates at random. These included: Trichophyton tonsurans (n = 125), Microsporum canis (n = 94), T. violaceum (n = 63) and M. audouinii (n = 19). The terbinafine minimum inhibitory concentration (MIC) range was 0.001-0.25 microg mL(-1), while MIC(50) and MIC(90) ranged between 0.002 and 0.125 microg mL(-1) and 0.03 and 0.25 microg mL(-1), respectively, for all species tested. MIC(50) and MIC(90) varied by individual species; however, there was no difference in terbinafine MIC among the different species isolated from U.S. and non-U.S. sites. CONCLUSION: Terbinafine demonstrates potent antifungal activity against dermatophyte isolates obtained from patients with tinea capitis worldwide. PMID- 18510669 TI - In vivo measurement of skin erythema and pigmentation: new means of implementation of diffuse reflectance spectroscopy with a commercial instrument. AB - BACKGROUND: Various physical, chemical and biological insults, including exposure to ultraviolet (UV) radiation, cause erythema and change in pigmentation in human skin. These reactions provide an important measure of the cutaneous response to the insult. OBJECTIVES: To present a new implementation of a method for objective in vivo measurement of erythema and pigmentation. METHODS: The method is based on acquisition of reflectance spectra in the visible range using a commercially available spectrophotometer. The probe of this instrument incorporates an integrating sphere that captures the light remitted from the skin in a wide range of angles. We corrected the acquired reflectance spectra for the contribution of specular reflections by an amount given by the Fresnel equation and verified this correction experimentally. This correction is particularly important when measurements are performed on heavily pigmented skin. The corrected reflectance spectra are then transformed into absorbance spectra. To analyse these spectra, we developed an algorithm which can be used to calculate apparent concentrations of oxyhaemoglobin, deoxyhaemoglobin and melanin. This method was tested in clinical studies of skin reactions induced by exposure to UV radiation. These experiments involved three groups of subjects with progressively darker complexion (constitutive pigmentation). Each group consisted of 10 subjects. Erythema was measured 1 day after UV exposure, and pigmentation (melanin content) 1 week later. Results Distinct apparent absorbance spectra were obtained for dark, intermediate and fair skin. There was good agreement between reconstructed spectra and experimental data at relevant wavelengths. Difference absorption spectra were able to show the dose dependence of UV-induced responses, and erythema and pigmentation values obtained by the spectroscopic method showed good correlation with those derived by subjective visual grading. CONCLUSIONS: The results demonstrate that the presented methodology provides an objective noninvasive way of measuring UV-induced reactions independently of the level of constitutive pigmentation. PMID- 18510670 TI - The chemistry and synergy of benzoyl peroxide with clindamycin. PMID- 18510671 TI - Squamous cell carcinoma arising within folliculitis decalvans. PMID- 18510672 TI - Identical p53 gene mutation in malignant proliferating trichilemmal tumour of the scalp and small cell carcinoma of the common bile duct: the necessity for therapeutic caution? PMID- 18510673 TI - Interaction between p53 codon 72 polymorphism and melanocortin 1 receptor variants on suntan response and cutaneous melanoma risk. AB - BACKGROUND: Ultraviolet (UV) radiation-induced p53 activation promotes cutaneous pigmentation by increasing transcriptional activity of pro-opiomelanocortin (POMC) in the skin. Induction of POMC/alpha-melanocyte-stimulating hormone (alpha MSH) activates the melanocortin 1 receptor (MC1R), resulting in skin pigmentation. The common p53 codon 72 polymorphism alters the protein's transcriptional activity, which may influence the UV radiation-induced tanning response. OBJECTIVES: We assessed the association of the p53 codon 72 polymorphism with tanning response, and its interaction with MC1R variants on tanning response and skin cancer risk. METHODS: The assessment was done in a nested case-control study within the Nurses' Health Study [219 melanoma cases, 286 squamous cell carcinoma (SCC) cases, 300 basal cell carcinoma (BCC) cases and 874 controls], and among controls from four nested case-control studies within the Nurses' Health Study. RESULTS: We found that the p53 Proline (Pro) allele was positively associated with childhood tanning response only among black/dark brown haired women. Compared with the Arginine/Arginine (Arg/Arg) genotype, odds ratios (ORs) of childhood tanning tendency for Arg/Pro and Pro/Pro genotypes were 1.59 (95% CI, 0.96-2.65) and 1.56 (95% CI, 0.55-4.40), respectively. The association between MC1R variants and childhood tanning tendency was similar in both p53 Arg/Arg genotype and Pro allele carriers (Arg/Pro or Pro/Pro). The association of the p53 Pro/Pro genotype with melanoma risk was strongest among women with light pigmentation, and with MC1R variants, with the joint risk categories having the highest overall risk. We did not observe such interaction for SCC and BCC. CONCLUSIONS: Our study suggests the involvement of the p53 codon 72 polymorphism in the skin tanning response and potential interaction with skin pigmentation on melanoma risk. Further work is needed to evaluate the association between p53 and its associated proteins and skin cancer risk. PMID- 18510674 TI - Evidence of high levels of anxiety and depression in polymorphic light eruption and their association with clinical and demographic variables. AB - BACKGROUND: Polymorphic light eruption (PLE) is a prevalent photosensitivity condition associated with psychological distress. Objective To examine patients with PLE for evidence of anxiety and depression, the influence of demographic and clinical variables, and the coping strategies used. METHODS: In a cross-sectional design, patients with PLE (n = 145) who had attended a hospital dermatology department completed validated questionnaires assessing anxiety, depression, social anxiety, and coping strategies. Clinical variables examined were: (i) number of months of the year affected by PLE; (ii) facial involvement; (iii) time taken for the rash to resolve; and (iv) whether the patient ever used steroids for their condition. RESULTS: Participants ranged in age from 16-78 (mean 44 years, SD 11.9), 81% female, with a mean age at onset of PLE of 28 years. Evidence of high levels of anxiety and depression was found in PLE, with 22% and 8% of patients scoring as probable cases for anxiety and depression, respectively. Higher levels of anxiety were associated with younger age of onset of PLE (r = -0.25, P < 0.01) and facial involvement (t = 2.84, P < 0.01), and depression was also associated with facial involvement (t = 3.60, P < 0.01). Furthermore, higher levels of depression and anxiety were associated with the use of maladaptive coping strategies, and depression was found to be the principal predictor of quality of life. CONCLUSIONS: High levels of anxiety and depression occur in PLE. Clinicians should be alert to the potential need for psychological management, particularly in patients with facial involvement and a younger age of onset of PLE. PMID- 18510675 TI - Noninfectious dermatological diseases associated with chronic exposure to mine tailings in a Peruvian district. AB - BACKGROUND: Mine tailings are metallic wastes which are deposited in the environment due to mining activity. Long-term exposure to these metals is harmful to human health. OBJECTIVE: To determine if chronic exposure to mine tailings constitutes a risk factor for the development of dermatological diseases in the district of San Mateo de Huanchor (Lima, Peru). METHODS: An observational case control study was carried out in the communities of Mayoc, Daza and Tamboraque (exposed to mine tailings, case group) located in the district of San Mateo de Huanchor, and also in the communities of Choccna and Caruya (not exposed to mine tailings, control group) located in the same district. Out of 230 adults, 121 were exposed and 109 were not exposed to mine tailings and out of 135 children, 71 were exposed and 64 were not exposed to mine tailings. RESULTS: In the adult group, 71% of the exposed cases had some noninfectious dermatological disease while in the nonexposed group the frequency was 34% [P < 0.001; odds ratio (OR) 5.40; 95% confidence interval (CI) 3.02-9.68]. A statistically significant difference between groups was found for arsenical dermatitis, nonpruritic papulovesicular eruption, atopic dermatitis, contact dermatitis, seborrhoeic dermatitis and xerosis. In the paediatric population, 71 exposed and 64 nonexposed children were evaluated. Sixty-nine per cent of the exposed group had some noninfectious dermatological disease vs. 30% in the nonexposed group (P < 0.001; OR 6.00; 95% CI 2.71-13.31). A statistically significant difference between groups was found for xerosis and atopic dermatitis. CONCLUSION: Chronic exposure to mine tailings represents a risk factor for development of noninfectious dermatological diseases in both adults and children. PMID- 18510676 TI - Beta-papillomaviruses and psoriasis: an intra-patient comparison of human papillomavirus carriage in skin and hair. AB - BACKGROUND: Human papillomaviruses (HPVs) of the beta genus (beta-PV), especially HPV5 and HPV36, are proposed to play a pathogenic role in psoriasis, but many previous studies have failed to control for potential confounders, including treatment. OBJECTIVES: To re-examine the relationship between beta-PV and psoriasis addressing limitations present in previous studies and analyse intra patient concordance for carriage of HPV. METHODS: Plucked eyebrow hairs and forearm skin scrapes were collected from 20 newly diagnosed, previously untreated adult patients with psoriasis and 23 normal controls. A combination of type specific and degenerate polymerase chain reaction methods was used to achieve comprehensive HPV DNA detection. RESULTS: The prevalence of HPV in hair and skin from psoriasis patients was higher than in controls (83.3% vs. 46.7%, respectively, P < 0.03 corrected for age and clustering). HPV5 or HPV36 were not over-represented. The profile of diverse beta-PV types was comparable in the two groups. Intra-patient concordance for HPV DNA at separate sites was high (P < 0.00001). CONCLUSIONS: Our data do not support a specific causal role for HPV5 or HPV36 in psoriasis, but suggest that psoriatic skin may be more permissive for viral presence than normal skin. High intra-patient concordance for specific HPV types at separate sites, together with the ubiquity of HPV DNA in normal human skin, suggests that an individual becomes colonized with a particular beta-PV profile presumably to the exclusion of other types. To what extent this HPV profile is then causal in the subsequent development of hyperproliferative skin disease is unknown. PMID- 18510677 TI - Fibroblast activation protein alpha identifies mesenchymal stromal cells from human bone marrow. AB - Mesenchymal stromal cells (MSCs) have gained widespread popularity in cell therapy, but their development into clinical products has been impeded by the scarcity of cell-specific markers. We previously explored transcriptome and membrane proteome of MSCs, from which fibroblast activation protein alpha (FAP) was recognized as a prime surface marker candidate. The present study showed that FAP was constitutively expressed on MSCs, but not on other cells. FAP immunoselection yielded homogeneous MSCs from cryopreserved bone marrow (BM). These results suggest that FAP serves as a surface protein marker that can singly define MSCs from BM and possibly from other sources. PMID- 18510678 TI - Platelet factor 4-heparin complexes trigger immune responses independently of the MyD88 pathway. PMID- 18510679 TI - IgG subclasses of anti-FVIII antibodies during immune tolerance induction in patients with hemophilia A. AB - The eradication of inhibitory antibodies in patients with haemophilia A can be accomplished by frequent administration of high or intermediate doses of factor VIII (FVIII), so-called immune tolerance induction (ITI). This study monitored the distribution of IgG subclasses of anti-FVIII antibodies during ITI. FVIII specific antibodies of subclass IgG1 were detected in all inhibitor patients tested, anti-FVIII IgG4 in 16, IgG2 in 10 and IgG3 in one of 20 patients analysed. Levels of anti-FVIII IgG1 and IgG4 correlated well with inhibitor titres as measured by Bethesda assay. In low-titre inhibitor patients, anti-FVIII antibodies consisted primarily of subclass IgG1 whereas, anti-FVIII antibodies of subclass IgG4 were more prominent in patients with high titre inhibitors who needed prolonged treatment or who failed ITI. Longitudinal analysis of 14 patients undergoing ITI revealed that the relative contribution of IgG subclasses was constant for most of the patients analysed. In two patients, the relative contribution of IgG4 increased during ITI. Overall, our findings document the distribution and dynamics of anti-FVIII IgG subclasses during ITI. Future studies will need to address whether monitoring the relative contribution of anti-FVIII subclasses IgG1 and IgG4 may be useful for the identification of patients who are at risk of failing ITI. PMID- 18510680 TI - Risk factors for early death in neonates with Down syndrome and transient leukaemia. AB - Transient leukaemia (TL) in neonates with Down syndrome (DS) is characterized by the transient appearance of blast cells in the peripheral blood that resolves spontaneously. Some TL patients die at an early age due to organ failure. Seventy DS patients with TL were studied retrospectively to identify clinical and laboratory characteristics associated with early death (<6 months of age). Sixteen of 70 patients (22.9%) died early. The main causes of death were organ failure, particularly hepatic and cardiopulmonary failure. On univariate analysis, early gestational age (EGA), high white blood cell (WBC) count (> or =100 x 10(9)/l), percentage of peripheral blasts, elevated aspartate transaminase (AST), elevated direct bilirubin (DB), and low Apgar score were significantly associated with poor survival. On multivariate analysis, EGA, WBC count, and DB were independent predictors of poor outcome. A simple risk stratification system combining EGA and WBC count was devised to predict poor outcome. Term infants (EGA > or = 37 weeks) whose WBC count was lower than 100 x 10(9)/l had the best outcome [7.7% (3/39) died early], while preterm infants (EGA < 37 weeks) whose WBC count was higher than 100 x 10(9)/l had the worst outcome [54.5% (6/11) died early]. This stratification system may be useful for identifying high-risk patients who need early therapeutic interventions. PMID- 18510681 TI - Antiphospholipid antibodies in adults with immune thrombocytopenic purpura. AB - To determine the clinical significance of antiphospholipid antibodies (aPL) in patients with immune thrombocytopenic purpura (ITP), anticardiolipin (aCL) (IgG and IgM) and lupus anticoagulant (LA) were sought at diagnosis in 215 ITP adults with platelets <50 x 10(9)/l. aPL (aCL and/or LA) were detected in 55 patients (26%): aCL alone in 39 (18%), aCL and LA in 15 (7%) and LA alone in one (0.5%). LA was significantly associated with high IgG-aCL levels (P = 0.001). Among age, sex, initial platelet count, bleeding score, acute or chronic ITP outcome, only younger age was significantly associated with LA-positivity (mean age 29 +/- 14 years vs. 45 +/- 20 years, P = 0.002). After a median follow-up of 31 months, 14/215 (7%) patients developed thrombosis (four arterial, 10 venous and/or pulmonary embolism); four of them (29%) had high aCL levels and LA. Multivariate analysis significantly associated thrombosis events only with age [hazard ratio (HR) = 1.6; 95% confidence interval (CI): 1.2-2.4], LA (HR: 9.9; 95% CI: 2.3 43.4) or high IgG-aCL level (HR: 7.5; 95% CI; 1.8-31.5). Although the thrombosis rate was low, the significant associations between thrombosis and LA or high aCL level suggest that aPL should be tested at ITP diagnosis. PMID- 18510682 TI - Acquired pure red cell aplasia: updated review of treatment. AB - Pure red cell aplasia (PRCA) is a syndrome characterized by a severe normocytic anaemia, reticulocytopenia, and absence of erythroblasts from an otherwise normal bone marrow. Primary PRCA, or secondary PRCA which has not responded to treatment of the underlying disease, is treated as an immunologically-mediated disease. Although vigorous immunosuppressive treatments induce and maintain remissions in a majority of patients, they carry an increased risk of serious complications. Corticosteroids were used in the treatment of PRCA and this has been considered the treatment of first choice although relapse is not uncommon. Cyclosporine A (CsA) has become established as one of the leading drugs for treatment of PRCA. However, common concerns have been the number of patients treated with CsA who achieve sustained remissions and the number that relapse. This article reviews the current status of CsA therapy and compares it to other treatments for diverse PRCAs. PMID- 18510683 TI - R-CHOP followed by consolidative autologous stem cell transplant and low dose rituxan maintenance therapy for advanced mantle cell lymphoma. PMID- 18510684 TI - Lessons from a Jehovah's Witness with 5q- syndrome: role of systemic immunosuppression and kinetics of recovery with lenalidomide from a life threatening anaemia. PMID- 18510685 TI - The use of ecarin chromogenic assay and prothrombinase induced clotting time in the monitoring of lepirudin for the treatment of heparin-induced thrombocytopenia. AB - Lepirudin (r-hirudin) is one of the two alternative anticoagulants licensed to treat patients with heparin-induced thrombocytopenia (HIT). Manufacturer's guidelines state that lepirudin should be monitored using the activated partial thromboplastin time (APTT) ratio. However, several studies have demonstrated a plateau effect of higher concentrations of lepirudin on APTT ratios and variable results when comparing different APTT reagents. This study compares APTT ratios (using two different APTT reagents) with two other commercially available methods for directly quantifying plasma lepirudin levels: ecarin chromogenic assay and prothrombinase-induced clotting time in 95 samples from five patients receiving lepirudin anticoagulation for HIT. PMID- 18510686 TI - Efficacy of alemtuzumab and gemcitabine in a patient with enteropathy-type T-cell lymphoma. PMID- 18510687 TI - Audit of the peri-delivery use of unfractionated heparin in women on therapeutic low-molecular weight heparin. AB - There is no evidence-based approach for the optimal management of peri-delivery anticoagulation in women receiving therapeutic dose of low-molecular weight heparin (LMWH) during pregnancy. Nevertheless, the maintenance of anticoagulation for the maximal period peri-delivery appears appropriate in women considered to be at high risk of venous or arterial thromboembolism. We developed a regimen based on fixed thromboprophylactic dose of unfractionated heparin (UFH) peri delivery and undertook an audit to evaluate the use and feasibility of this approach and any adverse events. Fixed intravenous thromboprophylactic dose of UFH (15,000 units/24 h) was commenced on the evening prior to a planned delivery [induction of labour or elective caesarean section (CS)], stopped 4 h predelivery and restarted 2-6 h postdelivery. Compliance was good with 32/38 consecutive deliveries managed according to the regimen. There were no cases of postpartum haemorrhage and no thrombosis associated with these 32 deliveries. Twenty-one patients were delivered by CS (11 elective) and eight patients received epidural/spinal anaesthesia without complication. In conclusion, the fixed thromboprophylactic dose UFH regimen provided maintenance of anticoagulation except for a matter of hours without excessive bleeding risk (conducive to neuroaxial anaesthesia) and was simple, flexible and acceptable to staff and patients. PMID- 18510688 TI - Myeloblastic transformation of chronic neutrophilic leukaemia. PMID- 18510689 TI - Malignant lymphoma and Schistosoma japonica infection. PMID- 18510690 TI - Von Willebrand disease and angiodysplasia responding to atorvastatin. PMID- 18510691 TI - Gemtuzumab ozogamicin as maintenance therapy after autologous stem cell transplantation in elderly patients with acute myeloid leukaemia. PMID- 18510692 TI - Platelet transfusion refractoriness. AB - The platelet, a fascinating anucleate cell, is critically important for haemostasis. Platelet transfusions have greatly reduced the incidence of major haemorrhagic complications associated with the management of haematological and oncological disorders. However, some patients fail to receive the full benefit of platelet transfusions because they do not achieve the appropriate platelet count increment following transfusion. This review will discuss the aetiology, diagnosis, and management of refractoriness to platelet transfusion, a complicated problem for both the treating physicians and the transfusion services supporting these patients. Although advances have been made in the diagnosis and treatment of immune-mediated platelet refractoriness, which is usually caused by anti-human leucocyte antigen antibodies, non-immune causes, such as sepsis, remain problematic. PMID- 18510693 TI - Neuro-mediators as predictors of paediatric atopic dermatitis. AB - BACKGROUND: Attempts to identify predictors of atopic dermatitis (AD) have focused on genetic and immunologic factors. However, the role of neuro-mediators remains to be elucidated. OBJECTIVE: To evaluate nerve growth factor (NGF) and vaso-active intestinal peptide (VIP) in predicting paediatric AD and assess their correlation with intrinsic and extrinsic types of AD. METHODS: We performed a nested case-control study in the prospective Taiwan birth panel cohort study. Cord and maternal plasma and questionnaires were gathered at birth. During follow up, we identified 40 available AD cases, which were matched to 80 unaffected controls chosen from this cohort. The concentrations of IgE, NGF, and VIP in cord and maternal plasma of these subjects were performed by ELISA. Receiver-operating characteristic (ROC) curves were generated to see how well each biomarker could predict AD. RESULTS: The NGF levels were significantly higher in AD patients than controls (mean+/-SD: 65.47+/-44.45 vs. 49.21+/-12.18 pg/mL for cord plasma and 89.68+/-41.04 vs. 66.96+/-23.05 pg/mL for maternal plasma) (P<0.05). VIP levels were also higher but not statistically significant. Plasma NGF may be a better biomarker than IgE in detecting paediatric AD (area under the ROC curve=0.65 vs. 0.61 for cord plasma and 0.69 vs. 0.61 for maternal plasma). Maternal NGF levels were significantly higher in patients with both intrinsic (96.18+/-48.15 pg/mL) and extrinsic (86.18+/-37.23 pg/mL) types of AD compared with controls (66.96+/ 23.05 pg/mL) (P<0.05). We assessed a significant correlation between self reported stress during pregnancy and maternal NGF levels (r=0.22, P=0.02). CONCLUSION: Our results suggest that NGF is a good alternative biomarker in predicting children with a risk of AD. PMID- 18510694 TI - Oral delivery of Lactobacillus casei Shirota modifies allergen-induced immune responses in allergic rhinitis. AB - BACKGROUND: Changes in the composition of the gut microbiota have been implicated in the pathogenesis of allergic disorders, suggesting beneficial interactions between the intestinal immune system and specific bacterial strains. Lactobacilli are naturally present within the complex gastrointestinal microbiota of humans and they are currently present in many probiotic supplements. OBJECTIVE: We sought to investigate the role that Lactobacillus casei Shirota (LcS) may play in modulating seasonal allergic rhinitis (SAR). METHODS: The study format was double blinded, placebo-controlled with 10 SAR sufferers in each group. We have documented and compared changes in immune status arising through the daily ingestion of a milk drink with or without live LcS, over a period of 5 months. Pre-, peak- and post-grass pollen season blood samples were collected for determination of plasma total IgE and grass pollen-specific IgG and IgE levels by an enzyme immunoassay. At the same time, cytokine levels were determined by flow cytometric bead array technology following culture of peripheral blood mononuclear cells for 6 days in the presence or absence of specific grass pollen antigens. RESULTS: Volunteers treated with LcS showed a significant reduction in levels of antigen-induced IL-5, IL-6 and IFN-gamma production compared with volunteers supplemented with placebo. Meanwhile, levels of specific IgG increased and IgE decreased in the probiotic group. CONCLUSION: Changes in antigen-induced production of cytokines were observed in patients treated with probiotics. These data show that probiotic supplementation modulates immune responses in allergic rhinitis and may have the potential to alleviate the severity of symptoms. PMID- 18510695 TI - Asthma induced by inhalation of flour in adults with food allergy to wheat. AB - BACKGROUND: Wheat is one of the major food allergens and it is also an inhalant allergen in workers exposed to flour dusts. Food allergy to wheat in adulthood seems to be rare and has never been reported to be associated with asthma induced by flour inhalation. OBJECTIVE: The study aimed at detecting adults with food allergy to wheat and screening them for the presence of specific bronchial reactivity to inhaled wheat proteins. METHODS: Adults with a history of adverse reactions to ingestion of wheat underwent skin prick test with commercial wheat extract and were assessed for the presence of specific wheat IgE in the sera. Food sensitivity to wheat was confirmed by double-blind, placebo-controlled food challenge (DBPCFC). Specific bronchial reactivity was investigated through a specific bronchial challenge with wheat proteins. RESULTS: In nine patients with evidence of specific IgE response to wheat, a diagnosis of food allergy was made by DBPCFC. Only two subjects had asthma as disease induced by ingestion of wheat. Seven subjects reported a history of respiratory symptoms when exposed to flour dusts. A significant reduction of forced expiratory volume in 1 s (FEV(1)) was detected in these seven patients when a specific bronchial challenge with flour proteins was performed. Only three out of seven subjects with asthma induced by flour could be considered occupationally exposed to flour dusts. CONCLUSION: For the first time, it has been shown that specific bronchial reactivity to wheat proteins can be detected in patients with different disorders associated with food allergy to wheat. The presence of asthma induced by inhaled flour is not strictly related to occupational exposure and it may also occur in subjects not displaying asthma among symptoms induced by wheat ingestion. PMID- 18510696 TI - A double-blind placebo-controlled birch allergy vaccination study II: correlation between inhibition of IgE binding, histamine release and facilitated allergen presentation. AB - BACKGROUND: The pathogenesis of IgE-mediated allergic disease is closely related to the production of T-helper type 2 (Th2) cytokines, which lead to IgE production pivotal for activation of mast cells and basophils. Proliferating T cells along with eosinophils expanded and attracted by Th2 cytokines are major contributors to the late-phase reaction. The activation of these Th2 cells is strongly enhanced by CD23-mediated IgE facilitated allergen presentation (FAP). OBJECTIVE: The present study aims to investigate the effect of specific immunotherapy (SIT)-induced allergen-specific non-IgE antibodies (blocking antibodies) on IgE binding to allergen, histamine release (HR) and CD23-mediated allergen uptake in antigen-presenting cells. METHODS: Competition between IgE and non-IgE for allergen binding was studied by Advia Centaur antibody measurements, passively sensitized basophils were used to study HR and IgE-facilitated binding of allergen to B cells (FAP) was studied by flow cytometry. FAP measurements were performed both with and without the addition of a reference IgE serum, which was included to obtain optimal complex formation. The serum samples were obtained from birch pollen immunotherapy (n=21) or placebo control patients (n=21) before and after 1 and 2 years of treatment. RESULTS: Statistically significant reduction of all parameters investigated was observed after 1 year of treatment and the effect was maintained during the second year of treatment. There was a clear correlation between the two FAP measurements and between each of them and the level of T cell activation reported upon previously. Moreover, strong correlations were found between changes in FAP, IgE binding and HR. CONCLUSION: The present study clearly demonstrates that SIT induces changes in the composition of serum antibodies that inhibit IgE binding, HR and FAP to a similar extent. This suggests that these measurements, individually or in combination, may be used to monitor the immunological effect of SIT, even though direct correlations to changes in clinical parameters could not be demonstrated. PMID- 18510697 TI - Foxp3 expression on normal and leukemic CD4+CD25+ T cells implicated in human T cell leukemia virus type-1 is inconsistent with Treg cells. AB - Foxp3 is a master gene of Treg cells, a novel subset of CD4(+) T cells primarily expressing CD25. We describe here different features in Foxp3 expression profile between normal and leukemic CD4(+)CD25(+) T cells, using peripheral blood samples from healthy controls (HCs), human T-cell leukemia virus type-1 (HTLV-1)-infected asymptomatic carriers (ACs), patients with adult T-cell leukemia (ATL), and various hematopoietic cell lines. The majority of CD4(+)CD25(+) T cells in HCs were positive for Foxp3, but not all CD4(+)CD25(+) T cells in ACs were positive, indicating that Foxp3 expression is not always linked to CD25 expression in normal T cells. Leukemic (ATL) T cells constitutively expressing CD25 were characteristic of heterogeneous Foxp3 expression, such as intra- and inter-case heterogeneity in intensity, inconsistency with CD25 expression, and a discrepancy in the mRNA and its protein expression. Surprisingly, a discernible amount of Foxp3 mRNA was detectable even in most cell lines without CD25 expression, a small fraction of which was positive for the Foxp3 proteins. The subcellular localization of Foxp3 in HTLV-1-infected cell lines was mainly cytoplasmic, different from that of primary ATL cells. These findings indicate that Foxp3 has two facets: essential Treg identity and molecular mimicry secondary to tumorigenesis. Conclusively, Foxp3 in normal T cells, but not mRNA, is basically potent at discriminating a subset of Treg cells from CD25(+) T-cell populations, whereas the modulation of Foxp3 expression in leukemic T cells could be implicated in oncogenesis and has a potentially useful clinical role. PMID- 18510698 TI - Study of gene expression profile during cord blood-associated megakaryopoiesis. AB - AIMS: To study the gene profile in cord blood (CB)-associated megakaryopoiesis. METHODS: In vitro differentiation of megakaryocytes (Mks) was carried out using human CB CD34(+) cells under the stimulation of recombinant human interleukin-3, stem cell factor and thrombopoietin for 7 d, followed by thrombopoietin only for further 3 d. Lineage-specific differentiation of Mk was examined by the expression of CD41 using flow cytometry and confocal microscopy. Total cellular RNA was extracted from day-0 CD34(+), day-10 CD41(+) and CD41(-) populations were isolated by immunomagnetic sorting respectively. Microarray was performed, and the data were analyzed using the GeneChip Operating System, Spotfire software and Genomatix BiblioSphere. RESULTS: Flow cytometric analysis showed 19.44 +/- 3.05% CD41(+) cells at day 10 of culture. The purity of CD41(+) population was enriched to 95.70 +/- 4.19% after sorting. Gene expression profiling revealed an upregulation of 285 and downregulation of 53 unique genes in the CD41(+) cells compared with CD41(-) and CD34(+) cells. Platelet-associated genes, such as thrombospondin 1, platelet glycoprotein IIIa, etc., were highly expressed in CD41(+) cells but not in CD41(-) cells and CD34(+) cells. Moreover, some genes that have not been reported to be associated with CB-derived megakaryopoiesis, such as Cbl-interacting proteins Sts-1, protocadherin 21, etc., are found to be highly expressed in the CD41(+) cells from this study. CONCLUSIONS: This study reveals a global gene expression profile of in vitro human CB-derived megakaryopoiesis at day 10. Some of these genes may play regulatory roles during the development of CB-derived megakaryopoiesis. PMID- 18510699 TI - Etoposide-initiated MLL rearrangements detected at high frequency in human primitive hematopoietic stem cells with in vitro and in vivo long-term repopulating potential. AB - Rearrangements initiating within the well-characterized break-point cluster region of the mixed lineage leukemia (MLL) gene on 11q23 are a hallmark of therapy-related leukemias following treatment with topoisomerase II poisons including etoposide. Hematopoietic stem cells (HSC) are believed to be the target cell for leukemia-initiating MLL rearrangement events. Although etoposide treatment is sufficient to induce readily detectable MLL rearrangements in primary human CD34+ cells, the majority of cells that gain translocations do not proliferate in culture possibly due to reduced proliferative capacity of most CD34+ cells during normal differentiation [Blood 2005;105:2124]. We characterized the impact of etoposide on primary human long-term repopulating HSC that represent only a minor portion of CD34+ cells. The proliferative capacity of HSC is dramatically increased following both a single and multiple exposures to etoposide as determined by their ability to engraft bone marrow of immune deficient non-obese diabetic/severe combined immunodeficient mice and to initiate hematopoiesis in long-term initiating cultures. Similar to results in CD34+ cells, a significant proportion of etoposide-treated HSC-derived clones harbored stable MLL rearrangements, including duplications, inversions and translocations. These results indicate HSC are highly susceptible to etoposide-induced and potentially oncogenic rearrangements initiating within MLL, and these HSC are particularly proficient for continued long-term proliferation both in vivo and in vitro. PMID- 18510700 TI - A phase I clinical trial of the histone deacetylase inhibitor belinostat in patients with advanced hematological neoplasia. AB - PURPOSE: To determine the safety, dose-limiting toxicity and maximum tolerated dose (MTD) of the novel hydroxamate histone deacetylase inhibitor belinostat (PXD101) in patients with advanced hematological neoplasms. PATIENTS AND METHODS: Sequential dose-escalating cohorts of three to six patients with hematological malignancies received belinostat administered as a 30-min i.v. infusion on days 1 5 of a 21-d cycle. Experience from a parallel dose-finding study in patients with solid tumors influenced the selection of the final dose. RESULTS: Sixteen patients received belinostat at one of three dose levels: 600 mg/m(2)/d (three patients), 900 mg/m(2)/d (three patients) and 1000 mg/m(2)/d (10 patients), the dose determined to be the MTD in a phase I solid tumor study [Steele et al. (2008) Clin Cancer Res, 14, 804-10]. The most common treatment-related adverse events (all grades) were nausea (50%), vomiting (31%), fatigue (31%) and flushing (31%). No grade 3 or 4 hematological toxicity compared with baseline occurred except one case of grade 3 lymphopenia. There were two related grade 4 adverse events of renal failure observed. Both events occurred in patients with multiple myeloma and had similar characteristics, i.e. an acute episode of decrease in renal function (pre-existing nephropathy in one patient), with a metabolic profile and decrease in tumor burden consistent with tumor lysis syndrome. No other related grade 4 events were noted. The only related grade 3 events noticed in more than one patient were fatigue and neurological symptoms (one patient had status epilepticus in association with uremia and one patient had paresthesia), all other related grade 3 events occurred in single patients. No cardiac events were noted. No complete or partial remissions were noted in these heavily pre treated (median of four prior regimens) patients. However, five patients, including two patients with diffuse large-cell lymphoma [including one patient with transformed chronic myelocytic leukaemia (CLL)], two patients with CLL and one patient with multiple myeloma, achieved disease stabilization in of two to nine treatment cycles. CONCLUSIONS: Intravenous belinostat at 600, 900 and 1000 mg/m(2)/d is well tolerated by patients with hematological malignancies. The study was carried out in parallel to a similar dose-finding study in patients with solid tumors, in which the MTD was determined to be 1000 mg/m(2)/d days 1-5 in a 21-d cycle. This dose can also be recommended for phase II studies in patients with hematological neoplasms. PMID- 18510701 TI - BK virus infection and neurologic dysfunctions in a patient with lymphoma treated with chemotherapy and rituximab. PMID- 18510702 TI - Rituximab therapy in adult patients with relapsed or refractory immune thrombocytopenic purpura: long-term follow-up results. AB - OBJECTIVE: To evaluate the long-term activity and toxicity profile of rituximab in adult patients with idiopathic immune thrombocytopenic purpura (ITP). PATIENTS AND METHODS: Twenty-six patients with active and symptomatic ITP relapsed or refractory received weekly infusions of rituximab 375 mg/m(2) for 4 wk. Median time from diagnosis to rituximab was 34.5 months. The following parameters of efficacy and toxicity were considered: complete response (CR) and partial response (PR), relapse rate, relapse-free survival (RFS), therapy-free survival (TFS), short- and long-term toxicity. RESULTS: CR and PR were 14/26 (54%) and 4/26 (15%), respectively. Median time of observation was 56.5 months (range 39 77). Nine of the 18 responding patients relapsed after a median of 21 months (range 8-66); 9/26 patients (35%) maintained the response, with a median follow up of 57 months (range 39-69), and 11/26 (42%) did not necessitate further therapy; estimated 5 yr RFS and TFS were 61% and 72%, respectively. Younger age and shorter interval from diagnosis to rituximab appeared indicators of better outcome. Rituximab administration was associated with two episodes of short-term toxicity, with one case of serum sickness syndrome; no infectious or other significant long-term complications were documented. CONCLUSION: Rituximab therapy may achieve long-lasting remission in nearly one-third of patients with relapsed or refractory ITP, with a good safety profile. PMID- 18510703 TI - Serum BAFF predicts prognosis better than APRIL in diffuse large B-cell lymphoma patients treated with rituximab plus CHOP chemotherapy. AB - OBJECTIVES: B-cell activating factor of the tumor necrosis factor family (BAFF) and a proliferation-inducing ligand (APRIL) regulate survival and proliferation of B cells. Thus the association of elevated serum levels of BAFF and APRIL with worse prognosis has been suggested in B-cell lymphoid malignancies. However, the prognostic relevance of BAFF and APRIL is unknown in patients treated with rituximab, a monoclonal antibody targeting B-cell depletion. METHODS: We measured serum levels of BAFF and APRIL by enzyme-linked immunosorbent assay in 66 patients newly diagnosed as diffuse large B-cell lymphoma (DLBCL). All patients were treated with rituximab-CHOP chemotherapy. RESULTS: The mean (+/-standard deviation) serum level of BAFF (1 970.21 +/- 1 979.45 pg/mL) was higher in DLBCL than in controls (861.03 +/- 194.92 pg/mL, Mann-Whitney U-test, P < 0.001). When the patients were dichotomized into high and low BAFF group based on the median value (1 258.00 pg/mL), high BAFF group had less numbers of complete responders to rituximab-CHOP, and more relapses or progression after or during treatment. In multivariate analysis, serum BAFF was an independent prognostic factor for overall survival and progression-free survival (P < 0.05). Although serum levels of APRIL was also higher than controls (10.60 +/- 19.08 ng/mL vs. 1.10 +/- 0.30 ng/mL, P = 0.023), it failed to show prognostic significance. CONCLUSIONS: Serum BAFF may be a useful indicator predicting prognosis in DLBCL patients treated with rituximab-containing chemotherapy. PMID- 18510704 TI - U-2973, a novel B-cell line established from a patient with a mature B-cell leukemia displaying concurrent t(14;18) and MYC translocation to a non-IG gene partner. AB - B-cell lymphomas/leukemias with simultaneous t(14;18)(q32;q21) and MYC rearrangements have recently been shown to constitute a separate diagnostic entity, presenting with a rapid clinical course and a very poor prognosis. We describe the establishment of an Epstein-Barr virus negative cell line, designated U-2973, from a male patient with a de novo aggressive B-cell lymphoma/leukemia and very high peripheral blast cell count. Flow cytometry of bone marrow cells and U-2973 displayed a mature B-cell phenotype, and immunostaining showed expression of MYC and BCL2. IG gene rearrangement data were consistent with a lymphoid neoplasm of germinal centre derivation. Cytogenetic studies using conventional G-banding, fluorescent in situ hybridization, spectral karyotyping and single nucleotide polymorphism array demonstrated a complex karyotype with both a t(14;18) and double translocations between MYC and a non-IG gene partner located at chromosome 12p12.1. PMID- 18510705 TI - Aggressive NK cell leukaemia after splenectomy: association with CD95-resistant memory T-cell proliferation and recalcitrant clinical course of haemophagocytic syndrome. AB - We describe a 44-yr-old Japanese woman with persistent polyclonal T-cell proliferation and recalcitrant clinical course of haemophagocytic syndrome (HPS). T cells bearing alphabeta T-cell receptors (TCR) expressed increased amounts of CD95 and of CD45RO, which are phenotypically memory T cells. The TCR repertoire was broad and diverse. Regardless of CD95 expression, these cells were resistant to CD95-mediated apoptosis. Aggressive natural killer cell leukaemia (ANKL) without an association with Epstein-Barr virus was detected 1 month after therapeutic splenectomy that followed 3 yr of immunosuppressive therapy against HPS. The immunophenotype of these leukaemia cells was CD56, CD16(dim), CD7, CD45RA and they expressed some CD2, CD8 and HLA-DR. Moreover, hyperdiploid clones with complex chromosomal abnormalities were also detected. Latent NK-cell malignancy seemed to cause the CD95-resistant memory T-cell proliferation and splenectomy resulted in overt ANKL progression. There should be careful consideration of the risks versus benefits of splenectomy in HPS, in light of the possibility of fatal leukaemia/lymphoma progression. PMID- 18510706 TI - Prepubertal social subjugation and anabolic androgenic steroid-induced aggression in male rats. AB - Abused children are more prone to abuse drugs, such as anabolic androgenic steroids (AAS), as teenagers and display violence as adults. AAS use has been linked with elevated aggression. Thus, exposure to child abuse and AAS may potentiate aggression. A social subjugation paradigm was used as an animal model of childhood abuse to determine whether prior subjugation increases AAS-induced aggression in male rats. Prepubertal gonadally intact male rats were exposed to social subjugation, a novel cage experience, or remained undisturbed in their home cages. Experimental males were socially subjugated by being placed in the home cage of an adult male. At puberty, both subjugated and nonsubjugated rats were injected with either the AAS testosterone or vehicle. AAS treatment continued for 5 weeks. Aggression was measured during the last week of AAS exposure. AAS was then discontinued. Aggression was again tested 12 weeks after AAS withdrawal. Aggression was tested under three conditions: (i) physical provocation of the experimental male; (ii) provocation of the intruder male; and (iii) without provocation. Both AAS-treated males and socially subjugated males displayed significantly more aggression than did controls. Elevated aggression by subjugated males was still present 17 weeks after social subjugation. AAS males also showed increased aggression 12 weeks after AAS withdrawal. However, exposure to both social subjugation and AAS had no long-term effects on aggression. The results of the present study indicate that social subjugation may have lasting consequences on the expression of adaptive social behaviours. PMID- 18510707 TI - Analysis of selected genes in neuroendocrine tumours: insulinomas and phaeochromocytomas. AB - Insulinomas and phaeochromocytomas are neuroendocrine tumours that may be either sporadic or manifestation of a familial cancer syndromes and are both derived from the neural crest. In the present study, gene components of different signalling pathways were investigated in sporadic human insulinomas and phaeochromocytomas to identify the responsible candidates. Ret and k-ras were tested for activating point mutations, and NF1, p53, BRCA1, nm23-H1, SDHB and SDHD for loss of heterozygosity (LOH). Twenty-two sporadic insulinomas and 15 phaeochromocytomas were analysed by the polymerase chain reaction using restriction fragment length polymorphism or dinucleotide repeat polymorphism methods. The results of our analysis demonstrate that the most frequent changes were point mutations of k-ras: 23% of insulinomas and 62% of phaeochromocytomas harboured k-ras mutations. The analysis also showed two phaeochromocytomas with point mutations of the ret oncogene. Only one insulinoma showed LOH of NF1, and another showed LOH of p53. Allelic loss of BRCA1 was detected in two insulinomas, and of nm23-H1 in another insulinoma. Allelic losses of the SDHB gene were present in two phaeochromocytoma and one insulinoma cases and allelic losses of SDHD were present in one phaeochromocytoma case. The changes observed in phaeochromocytomas were more homogenous and confined to k-ras and ret oncogenes, whereas insulinomas showed more heterogenic situation. Our findings may contribute to a better understanding of the genetic profile of neuroendocrine tumours. PMID- 18510708 TI - Effect of oestrogen receptor alpha and beta agonists on brain N-methyl-D aspartate receptors. AB - Previous studies have shown the oestradiol modulation of brain N-methyl-D aspartate (NMDA) receptors composed of the NR1/2B subunits. The contribution of oestrogen receptor subtypes in this oestradiol modulation of NMDA receptors and its subunits is not known. The following experiments investigated whether an oestrogenic receptor subtype is involved in the oestradiol effect on NMDA receptor specific binding and subunit mRNA levels. Ovariectomised Sprague-Dawley rats were treated 2 days after ovariectomy for 2 weeks with 17beta-oestradiol, an agonist for oestrogen receptor (ER)alpha 4,4',4''-(4-propyl-[1H]-pyrazole-1,3,5 triyl)trisphenol (PPT) or an agonist for ER beta 2,3-bis(4-hydroxyphenyl) propionitrile (DPN) and compared with control vehicle-treated ovariectomised and intact rats. Uterus weights, used as a peripheral measure of oestrogenic activity, decreased after ovariectomy and increased by oestradiol and PPT but not DPN treatment. In the hippocampal CA1 oriens and CA1 radiatum, [(3)H]Ro 25-6981 specific binding, a NMDA/NR2B ligand, was decreased in ovariectomised compared to intact rats and this was prevented by 17beta-oestradiol or PPT but not DPN treatments; a similar pattern was observed in the CA2/3 and dentate gyrus but did not reach statistical significance. In situ hybridisation of the mRNA of the NMDA/2B subunit in the hippocampus CA1, CA2/3 and dentate gyrus showed a decrease in ovariectomised rats compared to controls and this was also prevented by 17beta oestradiol and PPT but not DPN treatments. In cingulate and prefrontal cortices, ovariectomy increased [(3)H]Ro 25-6981 specific binding compared to intact controls, which was corrected by 17beta-oestradiol treatment but neither by PPT, nor DPN. In the cortical regions, the lack of effect of the ER alpha or ER beta agonist whereas 17beta-oestradiol was active, suggesting that the oestradiol modulation of cortical NMDA receptors requires both ERs or that this modulation does not involve ERs. In the hippocampus, the results obtained suggest an oestrogenic genomic modulation of NMDA receptors containing the NR2B subunit, implicating an ER alpha. PMID- 18510709 TI - Kisspeptin: a novel regulator of reproductive function. AB - The UK and international neuroendocrine community was deeply shocked and saddened the unbelievably premature death of Michael Harbuz in Bristol in 2006. Mick was a superb friend and colleague, and played a huge part in the development and activities of the British Neuroendocrine Group/British Society for Neuroendocrinology (BSN), serving as both Membership Secretary and Treasurer between 1999 and 2004. Mick was a leader in the field of neuroendocrine-immune interactions, and brought a great deal of charisma, humour and ability to meetings and conferences. He was also a passionate and committed supporter of the progress of young researchers and of their participation in neuroendocrine events. He recognised that today's postgraduate students and postdoctoral research fellows are tomorrow's neuroendocrine researchers, be it in academia, the health services or industry. To recognise Mick's great commitment to and enthusiasm for postgraduate education both in the University of Bristol and in the BSN, we decided to honour and remember him by instituting the 'Michael Harbuz Young Investigator Prize Lecture' to be delivered annually. Dr Waljit Dhillo from Imperial College London was the inaugural recipient of this award, and presented his lecture at the Annual Meeting of the BSN in Nottingham in September 2007, upon which this review is based. Recent evidence demonstrates that the neuropeptide kisspeptin and its receptor, GPR54, have a fundamental role in initiating the onset of puberty and are important in regulating reproductive function. This review discusses the evidence available from animals and humans demonstrating that kisspeptin potently stimulates the release of gonadotrophins by stimulating the release of gonadotrophin-releasing hormone and that a lack of kisspeptin or GPR54 results in reproductive failure. PMID- 18510710 TI - Genetic variation in Arabidopsis thaliana for night-time leaf conductance. AB - Night-time leaf conductance (g(night)) and transpiration may have several adaptive benefits related to plant water, nutrient and carbon relations. Little is known, however, about genetic variation in g(night) and whether this variation correlates with other gas exchange traits related to water use and/or native habitat climate. We investigated g(night) in 12 natural accessions and three near isogenic lines (NILs) of Arabidopsis thaliana. Genetic variation in g(night) was found for the natural accessions, and g(night) was negatively correlated with native habitat atmospheric vapour pressure deficit (VPD(air)), suggesting lower g(night) may be favoured by natural selection in drier habitats. However, there were also significant genetic correlations of g(night) with daytime gas exchange traits expected to affect plant fitness [i.e. daytime leaf conductance, photosynthesis and intrinsic water-use efficiency (WUE(i))], indicating that selection on daytime gas exchange traits may result in indirect selection on g(night). The comparison of three NILs to their parental genotypes identified one quantitative trait locus (QTL) contributing to variation in g(night). Further characterization of genetic variation in g(night) within and among populations and species, and of associations with other traits and native habitats will be needed to understand g(night) as a putatively adaptive trait. PMID- 18510711 TI - Contribution of FAT/CD36 to the regulation of skeletal muscle fatty acid oxidation: an overview. AB - Long chain fatty acids (LCFAs) are an important substrate for ATP production within the skeletal muscle. The process of LCFA delivery from adipose tissue to muscle mitochondria involves many regulatory steps. Recently, it has been recognized that LCFA oxidation is not only dependent on LCFA delivery to the muscle, but also on regulatory steps within the muscle. Increasing selected fatty acid binding proteins/transporters on the plasma membrane facilitates a very rapid LCFA increase into the muscle, independent of any changes in LCFA delivery to the muscle. Such a mechanism of LCFA transporter translocation is activated by muscle contraction. Intramuscular triacylglycerols may also be hydrolysed to provide fatty acids for mitochondrial oxidation, particularly during exercise, when hormone-sensitive lipase and other enzymes are activated. Mitochondrial LCFA entry is also highly regulated. This however does not involve only the malonyl CoA carnitine palmitoyltransferase-I (CPTI) axis. Exercise-induced fatty acid entry into mitochondria is also regulated by at least one of the proteins (FAT/CD36) that also regulates plasma membrane fatty acid transport. Among individuals, differences in mitochondrial fatty acid oxidation appear to be correlated with the content of mitochondrial CPTI and FAT/CD36. This paper provides a brief overview of mechanisms that regulate LCFA uptake and oxidation in skeletal muscle during exercise and in obesity. We focus largely on our own work on FAT/CD36, which contributes to regulating, in a coordinated fashion, LCFA uptake across the plasma membrane and the mitochondrial membrane. Very little is known about the roles of FATP1-6 on fatty acid transport in skeletal muscle. PMID- 18510712 TI - Method for electroporation for the early chick embryo. AB - In vitro whole-embryo culture of chick embryos, originally invented by New, has been widely used for studies of early embryogenesis. Here, a method for electroporation using the New culture and its derivatives is described, to achieve misexpression of exogenous gene in a temporally and spatially controlled manner in gastrulating chick embryos. Detailed information for the devices and procedures, and some experimental examples are presented. PMID- 18510713 TI - RNA interference by expressing short hairpin RNA in the Ciona intestinalis embryo. AB - We carried out RNA interference by expressing short hairpin RNA (shRNA) in the Ciona intestinalis embryo. For this purpose, we identified a gene encoding U6 small nuclear RNA (snRNA) in the C. intestinalis genome. The 1-kb sequence upstream of the U6 snRNA gene was sufficient for directing transcription of short RNA as revealed by Northern blot hybridization. An shRNA-expressing plasmid vector was constructed, in which shRNA-encoding oligonucleotides are inserted downstream of the U6 promoter. An shRNA that contained a sequence homologous to the C. intestinalis tyrosinase gene (Ci-tyrosinase) suppressed melanization of pigment cells in the brain of morphologically normal tailbud embryos. An shRNA that perfectly matched the translated sequence of enhanced green fluorescent protein (EGFP) (a mutant type of Aequorea victoria green fluorescent protein) suppressed the expression of the coelectroporated EGFP transgene. These results suggest that the expression of shRNA interferes with functions of both endogenous and exogenous genes. The shRNA-expressing plasmid constructed in the present study provides an easy and inexpensive alternative for the functional analysis of genes in ascidian embryos. PMID- 18510714 TI - Electroporation as an efficient method of gene transfer. AB - Gene transfer by electroporation is an indispensable method for the study of developmental biology, especially for the study using chick embryos. Here we briefly review the principles of the method, and its application to chick embryos. Methods of transient misexpression and long-term misexpression by retrovirus vector or transposon system, and knockdown by small interference RNA are reviewed. PMID- 18510715 TI - Abundance of thraustochytrids and bacteria in the equatorial Indian Ocean, in relation to transparent exopolymeric particles (TEPs). AB - Thraustochytrid protists are often abundant in coastal waters. However, their population dynamics and substrate preferences in the oceanic water column are poorly understood. We studied the abundance and distribution of thraustochytrids, bacteria and TEPs in the equatorial Indian Ocean waters during September 2003, October 2004 and September 2006. Thraustochytrids and bacteria were abundant, suggesting high biological productivity of the region. Thraustochytrids were positively related to bacteria during October 2004 but not at other times, suggesting overlapping or varying substrate preferences at different times. Thraustochytrid and bacteria were positively related to TEPs only in a few stations during October 2004, but were mostly positively related to TEPs generated from in situ water in a roller table experiment. TEPs from natural samples during October 2004 had a much greater affinity to the lectin Concanavalin A than to Limulin compared with those in September 2006 and from the roller tank experiments. The chemical composition of TEPs might explain their relationship with thraustochytrids. Thraustochytrids averaged a higher biomass than bacteria in two of the three cruises, but were less frequent and more patchily distributed compared with bacteria. PMID- 18510716 TI - The clinical relevance of duodenal intraepithelial lymphocyte counts in children treated for coeliac disease. PMID- 18510717 TI - Human serum amyloid A (SAA) and high sensitive C-reactive protein (hsCRP) in preterm newborn infants with nosocomial infections. AB - Human serum amyloid A (SAA) and high sensitive C-reactive protein (hsCRP) and their relation to suggestive nosocomial infections (NIs) were investigated in very preterm (VPT) newborn infants. In a retrospective analysis, information of suggestive NI was matched to levels of SAA and hsCRP in 224 serum samples from 72 VPT newborn infants. As a control group, 35 healthy-term newborn infants were chosen. Of the 224 serum samples, 145 samples were not associated with nosocomial infections. However, 79 were associated with NI: of these 79, 42 were found to be culture-proven NI. Trimmed mean (alpha= 0.05) levels for SAA and hsCRP in VPT newborn infants were higher than in control term newborn infants (1.74, 2.67 mg/L vs. 0.78, 0.16 mg/L; p = 0.01 and <0.0001, respectively), and higher in the NI group than in the non-NI group (5.14, 5.74 mg/L vs. 1.03, 1.18; p < 0.01 and <0.0001; respectively). The areas under the curve (AUC) for hsCRP, calculated from the receiver-operator characteristic (ROC) curves, was greater (0.816; 95% CI 0.759-0.864) than for SAA (0.610; 95% CI 0.543-0.675). CONCLUSION: Identifying and monitoring of bacterial and fungal infections in VPT might be further improved by the use of SAA and hsCRP. PMID- 18510718 TI - Multidimensional measure for gastroesophageal reflux disease (MM-GERD) symptoms in children: a population-based study. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) symptoms are very common in children with major presenting symptoms of abdominal pain, heartburn and regurgitation. The presence of GERD symptoms often result in an impaired health related quality of life for both the patients and their parents. Evaluation of children with GERD symptoms continues to challenge physicians due to the lack of a validated measure for GERD symptoms. AIMS: To develop and test a multidimensional measure for GERD symptoms in children and to evaluate the responses of the measure among children attending pediatric gastroenterology (GI) clinics. METHODS: We conducted a cross-sectional study that enrolled children with GERD symptoms from pediatric GI clinic. All children and parents received a standardized questionnaire concerning socio-economic parameters, GERD symptoms, duration, frequency, intensity and missed activities due to GERD symptoms. Each child and parent pair was interviewed by a physician to complete baseline information for the multidimensional measure that consisted of four scales: symptoms scale (10 items), pain intensity scale (3 items); disability scale (3 items) and satisfaction scale (2 items). RESULTS: One hundred and thirty-three children participated in the study; 59% girls, ages 4 to 18 years, mean age = 10 +/- 3, 50%, 10 years and younger. There was an excellent correlation between the four-scales measure among children 7 years and younger (R = 0.70, p = 0.0001) and children >7 years (R = 0.74, p = 0.0001). The inter-item consistency (Cronbach's co-efficient alpha) for the symptoms items, pain intensity items, disability items and satisfaction items were 0.71, 0.74, 0.78 and 0.60, respectively, demonstrating adequate reliability of the measure. CONCLUSION: Children with GERD symptoms have good responses to the multidimensional measure for GERD symptoms, showing that the measure performed well across populations. The measure is reliable and specific for assessing the symptoms of GERD in children and is an appropriate outcomes measure for clinical trials involving GERD symptoms in children. PMID- 18510719 TI - Relationship between metabolic and genomic diversity in sesame (Sesamum indicum L.). AB - BACKGROUND: Diversity estimates in cultivated plants provide a rationale for conservation strategies and support the selection of starting material for breeding programs. Diversity measures applied to crops usually have been limited to the assessment of genome polymorphism at the DNA level. Occasionally, selected morphological features are recorded and the content of key chemical constituents determined, but unbiased and comprehensive chemical phenotypes have not been included systematically in diversity surveys. Our objective in this study was to assess metabolic diversity in sesame by nontargeted metabolic profiling and elucidate the relationship between metabolic and genome diversity in this crop. RESULTS: Ten sesame accessions were selected that represent most of the genome diversity of sesame grown in India, Western Asia, Sudan and Venezuela based on previous AFLP studies. Ethanolic seed extracts were separated by HPLC, metabolites were ionized by positive and negative electrospray and ions were detected with an ion trap mass spectrometer in full-scan mode for m/z from 50 to 1000. Genome diversity was determined by Amplified Fragment Length Polymorphism (AFLP) using eight primer pair combinations. The relationship between biodiversity at the genome and at the metabolome levels was assessed by correlation analysis and multivariate statistics. CONCLUSION: Patterns of diversity at the genomic and metabolic levels differed, indicating that selection played a significant role in the evolution of metabolic diversity in sesame. This result implies that when used for the selection of genotypes in breeding and conservation, diversity assessment based on neutral DNA markers should be complemented with metabolic profiles. We hypothesize that this applies to all crops with a long history of domestication that possess commercially relevant traits affected by chemical phenotypes. PMID- 18510720 TI - In silico identification of genes involved in selenium metabolism: evidence for a third selenium utilization trait. AB - BACKGROUND: Selenium (Se) is a trace element that occurs in proteins in the form of selenocysteine (Sec) and in tRNAs in the form of selenouridine (SeU). Selenophosphate synthetase (SelD) is required for both utilization traits. However, previous research also revealed SelDs in two organisms lacking Sec and SeU, suggesting a possible additional use of Se that is dependent on SelD. RESULTS: In this study, we conducted comparative genomics and phylogenetic analyses to characterize genes involved in Se utilization. Candidate genes identified included SelA/SelB and YbbB that define Sec and SeU pathways, respectively, and NADH oxidoreductase that is predicted to generate a SelD substrate. In addition, among 227 organisms containing SelD, 10 prokaryotes were identified that lacked SelA/SelB and YbbB. Investigation of selD neighboring genes in these organisms revealed a SirA-like protein and two hypothetical proteins HP1 and HP2 that were strongly linked to a novel Se utilization. With these new signature proteins, 32 bacteria and archaea were found that utilized these proteins, likely as part of the new Se utilization trait. Metabolic labeling of one organism containing an orphan SelD, Enterococcus faecalis, with 75Se revealed a protein containing labile Se species that could be released by treatment with reducing agents, suggesting non-Sec utilization of Se in this organism. CONCLUSION: These studies suggest the occurrence of a third Se utilization trait in bacteria and archaea. PMID- 18510721 TI - Dynamics of pro-inflammatory and anti-inflammatory cytokine release during acute inflammation in chronic obstructive pulmonary disease: an ex vivo study. AB - BACKGROUND: Exacerbations of Chronic obstructive pulmonary disease (COPD) are an important cause of the morbidity and mortality associated with the disease. Strategies to reduce exacerbation frequency are thus urgently required and depend on an understanding of the inflammatory milieu associated with exacerbation episodes. Bacterial colonisation has been shown to be related to the degree of airflow obstruction and increased exacerbation frequency. The aim of this study was to asses the kinetics of cytokine release from COPD parenchymal explants using an ex vivo model of lipopolysaccharide (LPS) induced acute inflammation. METHODS: Lung tissue from 24 patients classified by the GOLD guidelines (7F/17M, age 67.9 +/- 2.0 yrs, FEV1 76.3 +/- 3.5% of predicted) and 13 subjects with normal lung function (8F,5M, age 55.6 +/- 4.1 yrs, FEV1 98.8 +/- 4.1% of predicted) was stimulated with 100 ng/ml LPS alone or in combination with either neutralising TNFalpha or IL-10 antibodies and supernatant collected at 1,2,4,6,24, and 48 hr time points and analysed for IL-1beta, IL-5, IL-6, CXCL8, IL-10 and TNFalpha using ELISA. Following culture, explants were embedded in glycol methacrylate and immunohistochemical staining was conducted to determine the cellular source of TNFalpha, and numbers of macrophages, neutrophils and mast cells. RESULTS: In our study TNFalpha was the initial and predictive cytokine released followed by IL-6, CXCL8 and IL-10 in the cytokine cascade following LPS exposure. The cytokine cascade was inhibited by the neutralisation of the TNFalpha released in response to LPS and augmented by the neutralisation of the anti-inflammatory cytokine IL-10. Immunohistochemical analysis indicated that TNFalpha was predominantly expressed in macrophages and mast cells. When patients were stratified by GOLD status, GOLD I (n = 11) and II (n = 13) individuals had an exaggerated TNFalpha responses but lacked a robust IL-10 response compared to patients with normal lung function (n = 13). CONCLUSION: We report on a reliable ex vitro model for the investigation of acute lung inflammation and its resolution using lung parenchymal explants from COPD patients. We propose that differences in the production of both TNFalpha and IL-10 in COPD lung tissue following exposure to bacterial LPS may have important biological implications for both episodes of exacerbation, disease progression and amelioration. PMID- 18510722 TI - Rasch fit statistics and sample size considerations for polytomous data. AB - BACKGROUND: Previous research on educational data has demonstrated that Rasch fit statistics (mean squares and t-statistics) are highly susceptible to sample size variation for dichotomously scored rating data, although little is known about this relationship for polytomous data. These statistics help inform researchers about how well items fit to a unidimensional latent trait, and are an important adjunct to modern psychometrics. Given the increasing use of Rasch models in health research the purpose of this study was therefore to explore the relationship between fit statistics and sample size for polytomous data. METHODS: Data were collated from a heterogeneous sample of cancer patients (n = 4072) who had completed both the Patient Health Questionnaire - 9 and the Hospital Anxiety and Depression Scale. Ten samples were drawn with replacement for each of eight sample sizes (n = 25 to n = 3200). The Rating and Partial Credit Models were applied and the mean square and t-fit statistics (infit/outfit) derived for each model. RESULTS: The results demonstrated that t-statistics were highly sensitive to sample size, whereas mean square statistics remained relatively stable for polytomous data. CONCLUSION: It was concluded that mean square statistics were relatively independent of sample size for polytomous data and that misfit to the model could be identified using published recommended ranges. PMID- 18510723 TI - Economic analysis including long-term risks and costs of alternative diagnostic strategies to evaluate patients with chest pain. AB - BACKGROUND: Diagnosis costs for cardiovascular disease waste a large amount of healthcare resources. The aim of the study is to evaluate the clinical and economic outcomes of alternative diagnostic strategies in low risk chest pain patients. METHODS: We evaluated direct and indirect downstream costs of 6 strategies: coronary angiography (CA) after positive troponin I or T (cTn-I or cTnT) (strategy 1); after positive exercise electrocardiography (ex-ECG) (strategy 2); after positive exercise echocardiography (ex-Echo) (strategy 3); after positive pharmacologic stress echocardiography (PhSE) (strategy 4); after positive myocardial exercise stress single-photon emission computed tomography with technetium Tc 99m sestamibi (ex-SPECT-Tc) (strategy 5) and direct CA (strategy 6). RESULTS: The predictive accuracy in correctly identifying the patients was 83,1% for cTn-I, 87% for cTn-T, 85,1% for ex-ECG, 93,4% for ex-Echo, 98,5% for PhSE, 89,4% for ex-SPECT-Tc and 18,7% for CA. The cost per patient correctly identified results $2.051 for cTn-I, $2.086 for cTn-T, $1.890 for ex ECG, $803 for ex-Echo, $533 for PhSE, $1.521 for ex-SPECT-Tc ($1.634 including cost of extra risk of cancer) and $29.673 for CA ($29.999 including cost of extra risk of cancer). The average relative cost-effectiveness of cardiac imaging compared with the PhSE equal to 1 (as a cost comparator), the relative cost of ex Echo is 1.5x, of a ex-SPECT-Tc is 3.1x, of a ex-ECG is 3.5x, of cTnI is x3.8, of cTnT is x3.9 and of a CA is 56.3x. CONCLUSION: Stress echocardiography based strategies are cost-effective versus alternative imaging strategies and the risk and cost of radiation exposure is void. PMID- 18510724 TI - Access to artemisinin combination therapy for malaria in remote areas of Cambodia. AB - BACKGROUND: Malaria-endemic countries are switching antimalarial drug policy to artemisinin combination therapies (ACTs) and the global community are considering the setting up of a global subsidy mechanism in order to make them accessible and affordable. However, specific interventions may be needed to reach remote at-risk communities and to ensure that they are used appropriately. This analysis documents the coverage with ACTs versus artemisinin monotherapies, and the effectiveness of malaria outreach teams (MOTs) and Village Malaria Workers (VMWs) in increasing access to appropriate diagnosis and treatment with ACTs in Cambodia, the first country to switch national antimalarial drug policy to an ACT of artesunate and mefloquine (A+M) in 2000. METHODS: A cross-sectional survey was carried out in three different types of intervention area: with VMWs, MOTs and no specific interventions. Individuals with a history of fever in the last three weeks were included in the study and completed a questionnaire on their treatment seeking and drug usage behaviour. Blood was taken for a rapid diagnostic test (RDT) and data on the household socio-economic status were also obtained. RESULTS: In areas without specific interventions, only 17% (42/251) of respondents received a biological diagnosis, 8% (17/206) of respondents who received modern drug did so from a public health facility, and only 8% of them (17/210) received A+M. Worryingly, 78% (102/131) of all artemisinin use in these areas was as a monotherapy. However, both the VMW scheme and MOT scheme significantly increased the likelihood of being seen by a trained provider (Adjusted Odds Ratios (AOR) of 148 and 4 respectively) and of receiving A+M (AORs of 2.7 and 7.7 respectively). CONCLUSION: The coverage rates of appropriate diagnosis and treatment of malaria were disappointingly low and the use of artemisinin monotherapy alarmingly high. This reflects the fragmented nature of Cambodia's health system in remote areas and the reliance placed by these communities on informal vendors from whom artemisinin monotherapies are widely available. However VMWs in particular are an effective means of improving access to malaria diagnosis and treatment. The VMW scheme and the social marketing of RDTS and blister-packaged artesunate and mefloquine have both been scaled up nationally. Case management in the public sector has also reportedly improved. Given recent concerns regarding the development of artemisinin drug resistance on the Thai-Cambodia border, the effectiveness of these measures in reducing the use of artemisinin monotherapy needs to be urgently re-evaluated. PMID- 18510725 TI - Surface-enhanced laser desorption/ionization time-of-flight proteomic profiling of breast carcinomas identifies clinicopathologically relevant groups of patients similar to previously defined clusters from cDNA expression. AB - INTRODUCTION: Microarray-based gene expression profiling represents a major breakthrough for understanding the molecular complexity of breast cancer. cDNA expression profiles cannot detect changes in activities that arise from post translational modifications, however, and therefore do not provide a complete picture of all biologically important changes that occur in tumors. Additional opportunities to identify and/or validate molecular signatures of breast carcinomas are provided by proteomic approaches. Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) offers high throughput protein profiling, leading to extraction of protein array data, calling for effective and appropriate use of bioinformatics and statistical tools. METHODS: Whole tissue lysates of 105 breast carcinomas were analyzed on IMAC 30 ProteinChip Arrays (Bio-Rad, Hercules, CA, USA) using the ProteinChip Reader Model PBS IIc (Bio-Rad) and Ciphergen ProteinChip software (Bio-Rad, Hercules, CA, USA). Cluster analysis of protein spectra was performed to identify protein patterns potentially related to established clinicopathological variables and/or tumor markers. RESULTS: Unsupervised hierarchical clustering of 130 peaks detected in spectra from breast cancer tissue lysates provided six clusters of peaks and five groups of patients differing significantly in tumor type, nuclear grade, presence of hormonal receptors, mucin 1 and cytokeratin 5/6 or cytokeratin 14. These tumor groups resembled closely luminal types A and B, basal and HER2 like carcinomas. CONCLUSION: Our results show similar clustering of tumors to those provided by cDNA expression profiles of breast carcinomas. This fact testifies the validity of the SELDI-TOF MS proteomic approach in such a type of study. As SELDI-TOF MS provides different information from cDNA expression profiles, the results suggest the technique's potential to supplement and expand our knowledge of breast cancer, to identify novel biomarkers and to produce clinically useful classifications of breast carcinomas. PMID- 18510726 TI - Meta-analysis confirms BCL2 is an independent prognostic marker in breast cancer. AB - BACKGROUND: A number of protein markers have been investigated as prognostic adjuncts in breast cancer but their translation into clinical practice has been impeded by a lack of appropriate validation. Recently, we showed that BCL2 protein expression had prognostic power independent of current used standards. Here, we present the results of a meta-analysis of the association between BCL2 expression and both disease free survival (DFS) and overall survival (OS) in female breast cancer. METHODS: Reports published in 1994-2006 were selected for the meta-analysis using a search of PubMed. Studies that investigated the role of BCL2 expression by immunohistochemistry with a sample size greater than 100 were included. Seventeen papers reported the results of 18 different series including 5,892 cases with an average median follow-up of 92.1 months. RESULTS: Eight studies investigated DFS unadjusted for other variables in 2,285 cases. The relative hazard estimates ranged from 0.85 - 3.03 with a combined random effects estimate of 1.66 (95%CI 1.25 - 2.22). The effect of BCL2 on DFS adjusted for other prognostic factors was reported in 11 studies and the pooled random effects hazard ratio estimate was 1.58 (95%CI 1.29-1.94). OS was investigated unadjusted for other variables in eight studies incorporating 3,910 cases. The hazard estimates ranged from 0.99-4.31 with a pooled estimate of risk of 1.64 (95%CI 1.36-2.0). OS adjusted for other parameters was evaluated in nine series comprising 3,624 cases and the estimates for these studies ranged from 1.10 to 2.49 with a pooled estimate of 1.37 (95%CI 1.19-1.58). CONCLUSION: The meta analysis strongly supports the prognostic role of BCL2 as assessed by immunohistochemistry in breast cancer and shows that this effect is independent of lymph node status, tumour size and tumour grade as well as a range of other biological variables on multi-variate analysis. Large prospective studies are now needed to establish the clinical utility of BCL2 as an independent prognostic marker. PMID- 18510727 TI - Associations between disease severity, coping and dimensions of health-related quality of life in patients admitted for elective coronary angiography - a cross sectional study. AB - BACKGROUND: In patients with suspected coronary artery disease (CAD), the overall aim was to analyse the relationships between disease severity and both mental and physical dimensions of health related quality of life (HRQOL) using a modified version of the Wilson and Cleary model. METHODS: Using a cross-sectional design, 753 patients (74% men), mean age 62 years, referred for elective cardiac catheterisation were included. The measures included 1) physiological factors 2) symptoms (disease severity, self-reported symptoms, anxiety and depression 3) self-reported functional status, 4) coping, 5) perceived disease burden, 6) general health perception and 7) overall quality of life. To analyse relationships, we performed linear and ordinal logistic regressions. RESULTS: CAD and left ventricular ejection fraction (LVEF) were significantly associated with symptoms of angina pectoris and dyspnea. CAD was not related to symptoms of anxiety and depression, but less depression was found in patients with low LVEF. Angina pectoris and dyspnea were both associated with impaired physical function, and dyspnea was also negatively related to social function. Overall, less perceived burden and better overall QOL were observed in patients using more confronting coping strategy. CONCLUSION: The present study demonstrated that data from cardiac patients to a large extent support the suggested model by Wilson and Cleary. PMID- 18510728 TI - Prediction of structural stability of short beta-hairpin peptides by molecular dynamics and knowledge-based potentials. AB - BACKGROUND: The structural stability of peptides in solution strongly affects their binding affinities and specificities. Thus, in peptide biotechnology, an increase in the structural stability is often desirable. The present work combines two orthogonal computational techniques, Molecular Dynamics and a knowledge-based potential, for the prediction of structural stability of short peptides (< 20 residues) in solution. RESULTS: We tested the new approach on four families of short beta-hairpin peptides: TrpZip, MBH, bhpW and EPO, whose structural stabilities have been experimentally measured in previous studies. For all four families, both computational techniques show considerable correlation (r > 0.65) with the experimentally measured stabilities. The consensus of the two techniques shows higher correlation (r > 0.82). CONCLUSION: Our results suggest a prediction scheme that can be used to estimate the relative structural stability within a peptide family. We discuss the applicability of this predictive approach for in-silico screening of combinatorial peptide libraries. PMID- 18510729 TI - Variation in expression of HMW1 and HMW2 adhesins in invasive nontypeable Haemophilus influenzae isolates. AB - BACKGROUND: Among surface antigens of nontypeable Haemophilus influenzae (NTHi), the HMW1 and HMW2 proteins are the major adhesins promoting colonization of the upper respiratory tract. Since they are potential vaccine candidates, knowledge concerning variation in HMW proteins expression among clinical isolates is of great interest. In this study, expression of hmw1A and hmw2A genes was evaluated by quantitative real-time reverse transcription-PCR in 3 NTHi invasive isolates (strains 56, 72, 91) and in the prototype strain 12. Number of 7-bp repeats within the hmwA promoters and presence of HMW proteins by Western blotting were also determined. RESULTS: Results showed that gene transcription varied not only among different isolates but also between the hmw1A and hmw2A genes from the same isolate. Compared to that found in prototype strain 12, up-regulation of the hmw1A gene expression was found in strain 56, down-regulation of both hmw1A and hmw2A genes transcripts was observed in strain 72 whereas the two hmwA genes appeared differentially expressed in strain 91 with the hmw1A transcript enhanced but the hmw2A transcript reduced. CONCLUSION: Increasing numbers of 7-bp repeats within the hmwA promoters generally correlated with decreased amounts of mRNA transcript, however additional control mechanisms contributing to modulation of hmw1A gene seem to be present. PMID- 18510730 TI - Impact of case management by advanced practice nurses in primary care on unplanned hospital admissions: a controlled intervention study. AB - BACKGROUND: Increasing unplanned hospital admissions disrupt planned health care, lead to additional morbidity and are expensive. A recent review found only weak evidence for case management preventing unplanned admissions, yet case management of older people is being implemented widely in the UK. We aimed to study the effect of advanced practice nurse case management on unplanned medical and geriatric hospital admission rates in patients 50 years and over, and on admission risk in a 'higher risk' sub-group of patients in the UK. METHODS: Case management by advanced practice nurses in NHS primary care practices in the Swansea Local Health Board area, Wales, UK. We conducted a prospective non randomized controlled intervention study comparing unplanned medical and geriatric patient admissions between five intervention and thirty non intervention practices during a pre-intervention year and an intervention year. RESULTS: For all lengths of stay, comparing intervention (n = 5) with non intervention practices (n = 30) from pre-intervention to intervention year, we found that the unplanned medical and geriatric admission rate was significantly lower in the intervention group - adjusted relative risk of 0.909; relative risk reduction 9.1% (95% credible limit 0.840 to 0.984, p = 0.018); absolute risk reduction 0.99 admissions per 100 patients (95% credible limit 0.17 to 1.86, p = 0.018). For lengths of stay of one night or more we observed a stronger effect - adjusted relative risk 0.896; relative risk reduction 10.41% (95%, credible limit 0.820 to 0.979, p = 0.015). Most of the rate reduction was due to a reduction in the number of new admissions but much less so for admissions of lengths of stay of at least one night, compared to all lengths of stay. We did not find a statistically significant effect on re-admission or multiple re-admission rates in 'higher risk' patients previously admitted one or more times - adjusted relative risk of further multiple admissions per previously admitted patient 0.908 (95% credible limit 0.765, 1.077); relative risk reduction 9.3%; adjusted relative risk of total admissions per multiple admitter 0.995 (95% credible limit 0.940, 1.053) relative risk reduction 0.6%. CONCLUSION: Although this study reports a reduction in unplanned admission rates in the intervention practices, this appears to be only in part directly due to nurse case management: most of the reduction did not occur in multipe admitters whom were case managed. Further research is needed to explain this finding, to elucidate how best to target the attention of case managers and to examine the complexity of potential outcomes in terms of the nature and necessity of admissions and most suitable lengths-of-stay in terms of acute care or rehabilittion need. PMID- 18510731 TI - Endothelial nitric oxide synthase gene polymorphisms and cardiovascular damage in hypertensive subjects: an Italian case-control study. AB - BACKGROUND: Nitric oxide (NO) synthesized by endothelial nitric oxide synthase (eNOS) plays an important role in regulation of endothelial function and in the control of blood pressure. However, the results from some studies on the association between three clinically relevant eNOS gene polymorphisms (G894T, T786C and intron 4b/a) and essential hypertension are unclear. We designed a case control study to evaluate the influence of eNOS polymorphisms on target organ damage in 127 hypertensives and 67 normotensives. Clinical evaluation, biochemical parameters, Urinary Albumin Excretion (UAE) and echocardiogram were performed to characterize target organ damage. eNOS polymorphism were recognized by PCR method. RESULTS: The distribution of eNOS genotypes was similar in hypertensives and normotensives but 4aa was present in the 2.5% of hypertensives and completely absent in normotensives. Subjects with 4bb, G894T, and T786C genotypes showed an increased prevalence of target organ damage. Moreover prevalence of G894T and introne 4 variants was significantly higher in hypertensives than in normotensives both with cardiovascular damage. Logistic regression analysis didn't show any association between eNOS polymorphisms, Body Mass Index (BMI), hypertension, gender and cardiovascular damage. Only the age (OR 1.11; IC 95% 1.06-1.18) was predictive of cardiovascular damage in our population. CONCLUSION: Our results seem to indicate a lack of association with eNOS variants and cardiovascular damage onset. PMID- 18510732 TI - ParaHox gene expression in larval and postlarval development of the polychaete Nereis virens (Annelida, Lophotrochozoa). AB - BACKGROUND: Transcription factors that encode ANTP-class homeobox genes play crucial roles in determining the body plan organization and specification of different organs and tissues in bilaterian animals. The three-gene ParaHox family descends from an ancestral gene cluster that existed before the evolution of the Bilateria. All three ParaHox genes are reported from deuterostomes and lophotrochozoans, but not to date from any ecdysozoan taxa, and there is evidence that the ParaHox genes, like the related Hox genes, were ancestrally a single chromosomal cluster. However, unlike the Hox genes, there is as yet no strong evidence that the ParaHox genes are expressed in spatial and temporal order during embryogenesis. RESULTS: We isolated fragments of the three Nereis virens ParaHox genes, then used these as probes for whole-mount in situ hybridization in larval and postlarval worms. In Nereis virens the ParaHox genes participate in antero-posterior patterning of ectodermal and endodermal regions of the digestive tract and are expressed in some cells in the segment ganglia. The expression of these genes occurs in larval development in accordance with the position of these cells along the main body axis and in postlarval development in accordance with the position of cells in ganglia along the antero-posterior axis of each segment. In none of these tissues does expression of the three ParaHox genes follow the rule of temporal collinearity. CONCLUSION: In Nereis virens the ParaHox genes are expressed during antero-posterior patterning of the digestive system (ectodermal foregut and hindgut, and endodermal midgut) of Nereis virens. These genes are also expressed during axial specification of ventral neuroectodermal cell domains, where the expression domains of each gene are re-iterated in each neuromere except for the first parapodial segment. These expression domains are probably predetermined and may be directed on the antero-posterior axis by the Hox genes, whose expression starts much earlier during embryogenesis. Our results support the hypothesis that the ParaHox genes are involved in antero-posterior patterning of the developing embryo, but they do not support the notion that these genes function only in the patterning of endodermal tissues. PMID- 18510733 TI - cpRAS: a novel circularly permuted RAS-like GTPase domain with a highly scattered phylogenetic distribution. AB - A recent systematic survey suggested that the YRG (or YawG/YlqF) family with the G4-G5-G1-G2-G3 order of the conserved GTPase motifs represents the only possible circularly permuted variation of the canonical GTPase structure. Here we show that a different circularly permuted GTPase domain actually does exist, conforming to the pattern G3-G4-G5-G1-G2. The domain, dubbed cpRAS, is a variant of RAS family GTPases and occurs in two types of larger proteins, either inserted into a region homologous to a bacterial group of proteins classified as COG2373 and potentially related to the alpha-2-macroglobulin family (so far a single protein in Dictyostelium) or in combination with a von Willebrand factor type A (VWA) domain. For the latter protein type, which was found in a few metazoans and several distantly related protists, existence in the common ancestor of opisthokonts, Amoebozoa and excavates followed by at least eight independent losses may be inferred. Our findings thus bring further evidence for the importance of parallel reduction of ancestral complexity in the eukaryotic evolution. PMID- 18510734 TI - Neuromyelitis optica pathogenesis and aquaporin 4. AB - Neuromyelitis optica (NMO) is a severe, debilitating human disease that predominantly features immunopathology in the optic nerves and the spinal cord. An IgG1 autoantibody (NMO-IgG) that binds aquaporin 4 (AQP4) has been identified in the sera of a significant number of NMO patients, as well as in patients with two related neurologic conditions, bilateral optic neuritis (ON), and longitudinal extensive transverse myelitis (LETM), that are generally considered to lie within the NMO spectrum of diseases. NMO-IgG is not the only autoantibody found in NMO patient sera, but the correlation of pathology in central nervous system (CNS) with tissues that normally express high levels of AQP4 suggests NMO IgG might be pathogenic. If this is the case, it is important to identify and understand the mechanism(s) whereby an immune response is induced against AQP4. This review focuses on open questions about the "events" that need to be understood to determine if AQP4 and NMO-IgG are involved in the pathogenesis of NMO. These questions include: 1) How might AQP4-specific T and B cells be primed by either CNS AQP4 or peripheral pools of AQP4? 2) Do the different AQP4 expressing tissues and perhaps the membrane structural organization of AQP4 influence NMO-IgG binding efficacy and thus pathogenesis? 3) Does prior infection, genetic predisposition, or underlying immune dysregulation contribute to a confluence of events which lead to NMO in select individuals? A small animal model of NMO is essential to demonstrate whether AQP4 is indeed the incipient autoantigen capable of inducing NMO-IgG formation and NMO. If the NMO model is consistent with the human disease, it can be used to examine how changes in AQP4 expression and blood-brain barrier (BBB) integrity, both of which can be regulated by CNS inflammation, contribute to inductive events for anti-AQP4 specific immune response. In this review, we identify reagents and experimental questions that need to be developed and addressed to enhance our understanding of the pathogenesis of NMO. Finally, dysregulation of tolerance associated with autoimmune disease appears to have a role in NMO. Animal models would allow manipulation of hormone levels, B cell growth factors, and other elements known to increase the penetrance of autoimmune disease. Thus an AQP4 animal model would provide a means to manipulate events which are now associated with NMO and thus demonstrate what set of events or multiplicity of events can push the anti-AQP4 response to be pathogenic. PMID- 18510736 TI - Malakoplakia of the appendix, an uncommon entity at an unusual site: a case report. AB - INTRODUCTION: Malakoplakia is an uncommon inflammatory condition usually affecting the genitourinary tract, which has been associated with infections, tumours and immunocompromised states. CASE PRESENTATION: We report a case of malakoplakia in the appendix of a 61-year-old man with a long-standing history of ulcerative colitis. Clinically and macroscopically malakoplakia can simulate tumours or abscesses and can cause diagnostic difficulties. Histologically malakoplakia in the gastrointestinal tract must be differentiated from Whipple disease, other infectious and noninfectious granulomatous disorders and histiocyte storage diseases. To the best of our knowledge, this is the first case of malakoplakia of the appendix reported in association with ulcerative colitis and the sixth reported case of malakoplakia of the appendix in the literature. Although the underlying disease in our case was ulcerative colitis, the malakoplakia was limited to the appendix. CONCLUSION: The significance of this finding is not clear but we feel that this was a localised manifestation of the underlying immunosuppressive state. Ulcerative colitis and treatment with steroids may make a patient immunosuppressive and the local and systemic change in the immunity may facilitate the proliferation of the organisms and modify the phagocytic abilities of the macrophages. PMID- 18510735 TI - Oxytocin-induced antinociception in the spinal cord is mediated by a subpopulation of glutamatergic neurons in lamina I-II which amplify GABAergic inhibition. AB - BACKGROUND: Recent evidence suggests that oxytocin (OT), secreted in the superficial spinal cord dorsal horn by descending axons of paraventricular hypothalamic nucleus (PVN) neurons, produces antinociception and analgesia. The spinal mechanism of OT is, however, still unclear and requires further investigation. We have used patch clamp recording of lamina II neurons in spinal cord slices and immunocytochemistry in order to identify PVN-activated neurons in the superficial layers of the spinal cord and attempted to determine how this neuronal population may lead to OT-mediated antinociception. RESULTS: We show that OT released during PVN stimulation specifically activates a subpopulation of lamina II glutamatergic interneurons which are localized in the most superficial layers of the dorsal horn of the spinal cord (lamina I-II). This OT-specific stimulation of glutamatergic neurons allows the recruitment of all GABAergic interneurons in lamina II which produces a generalized elevation of local inhibition, a phenomenon which might explain the reduction of incoming Adelta and C primary afferent-mediated sensory messages. CONCLUSION: Our results obtained in lamina II of the spinal cord provide the first clear evidence of a specific local neuronal network that is activated by OT release to induce antinociception. This OT-specific pathway might represent a novel and interesting therapeutic target for the management of neuropathic and inflammatory pain. PMID- 18510737 TI - Use of intravitreal bevacizumab in a patient with a Von Hippel-Lindau-associated retinal haemangioblastoma of the optic nerve head: a case report. AB - INTRODUCTION: The optimum management of a capillary haemangioblastoma affecting the optic nerve head is not clear. A number of treatment modalities have been used to treat the tumours and their consequences. Ocular haemangioblastomas express high levels of vascular endothelial growth factor and levels have been correlated with tumour growth and activity. Treatment with vascular endothelial growth factor inhibitors would therefore seem a logical approach. CASE PRESENTATION: We describe a 23-year-old man with an exophytic capillary haemangioblastoma of the optic nerve head that was treated with intravitreal bevacizumab injections. CONCLUSION: Unfortunately, treatment with intravitreal bevacizumab on three occasions had no effect on either tumour size or exudation in this patient. PMID- 18510738 TI - Identification of SNPs and INDELS in swine transcribed sequences using short oligonucleotide microarrays. AB - BACKGROUND: Genome-wide detection of single feature polymorphisms (SFP) in swine using transcriptome profiling of day 25 placental RNA by contrasting probe intensities from either Meishan or an occidental composite breed with Affymetrix porcine microarrays is presented. A linear mixed model analysis was used to identify significant breed-by-probe interactions. RESULTS: Gene specific linear mixed models were fit to each of the log2 transformed probe intensities on these arrays, using fixed effects for breed, probe, breed-by-probe interaction, and a random effect for array. After surveying the day 25 placental transcriptome, 857 probes with a q-value < or = 0.05 and |fold change| > or = 2 for the breed-by probe interaction were identified as candidates containing SFP. To address the quality of the bioinformatics approach, universal pyrosequencing assays were designed from Affymetrix exemplar sequences to independently assess polymorphisms within a subset of probes for validation. Additionally probes were randomly selected for sequencing to determine an unbiased confirmation rate. In most cases, the 25-mer probe sequence printed on the microarray diverged from Meishan, not occidental crosses. This analysis was used to define a set of highly reliable predicted SFPs according to their probability scores. CONCLUSION: By applying a SFP detection method to two mammalian breeds for the first time, we detected transition and transversion single nucleotide polymorphisms, as well as insertions/deletions which can be used to rapidly develop markers for genetic mapping and association analysis in species where high density genotyping platforms are otherwise unavailable.SNPs and INDELS discovered by this approach have been publicly deposited in NCBI's SNP repository dbSNP. This method is an attractive bioinformatics tool for uncovering breed-by-probe interactions, for rapidly identifying expressed SNPs, for investigating potential functional correlations between gene expression and breed polymorphisms, and is robust enough to be used on any Affymetrix gene expression platform. PMID- 18510739 TI - Exercise training improves relaxation response and SOD-1 expression in aortic and mesenteric rings from high caloric diet-fed rats. AB - BACKGROUND: Obesity has been associated with a variety of disease such as type II diabetes mellitus, arterial hypertension and atherosclerosis. Evidences have shown that exercise training promotes beneficial effects on these disorders, but the underlying mechanisms are not fully understood. The aim of this study was to investigate whether physical preconditioning prevents the deleterious effect of high caloric diet in vascular reactivity of rat aortic and mesenteric rings. METHODS: Male Wistar rats were divided into sedentary (SD); trained (TR); sedentary diet (SDD) and trained diet (TRD) groups. Run training (RT) was performed in sessions of 60 min, 5 days/week for 12 weeks (70-80% VO2max). Triglycerides, glucose, insulin and nitrite/nitrate concentrations (NOx-) were measured. Concentration-response curves to acetylcholine (ACh) and sodium nitroprusside (SNP) were obtained. Expression of Cu/Zn superoxide dismutase (SOD 1) was assessed by Western blotting. RESULTS: High caloric diet increased triglycerides concentration (SDD: 216 +/- 25 mg/dl) and exercise training restored to the baseline value (TRD: 89 +/- 9 mg/dl). Physical preconditioning significantly reduced insulin levels in both groups (TR: 0.54 +/- 0.1 and TRD: 1.24 +/- 0.3 ng/ml) as compared to sedentary animals (SD: 0.87 +/- 0.1 and SDD: 2.57 +/- 0.3 ng/ml). On the other hand, glucose concentration was slightly increased by high caloric diet, and RT did not modify this parameter (SD: 126 +/- 6; TR: 140 +/- 8; SDD: 156 +/- 8 and TRD 153 +/- 9 mg/dl). Neither high caloric diet nor RT modified NOx- levels (SD: 27 +/- 4; TR: 28 +/- 6; SDD: 27 +/- 3 and TRD: 30 +/- 2 microM). Functional assays showed that high caloric diet impaired the relaxing response to ACh in mesenteric (about 13%), but not in aortic rings. RT improved the relaxing responses to ACh either in aortic (28%, for TR and 16%, to TRD groups) or mesenteric rings (10%, for TR and 17%, to TRD groups) that was accompanied by up-regulation of SOD-1 expression and reduction in triglycerides levels. CONCLUSION: The improvement in endothelial function by physical preconditioning in mesenteric and aortic arteries from high caloric fed-rats was directly related to an increase in NO bioavailability to the smooth muscle mostly due to SOD-1 up regulation. PMID- 18510740 TI - Influence of acute pancreatitis on the in vitro responsiveness of rat mesenteric and pulmonary arteries. AB - BACKGROUND: Acute pancreatitis is an inflammatory disease characterized by local tissue injury and systemic inflammatory response leading to massive nitric oxide (NO) production and haemodynamic disturbances. Therefore, the aim of this work was to evaluate the vascular reactivity of pulmonary and mesenteric artery rings from rats submitted to experimental pancreatitis. Male Wistar rats were divided into three groups: saline (SAL); tauracholate (TAU) and phospholipase A2 (PLA2). Pancreatitis was induced by administration of TAU or PLA2 from Naja mocambique mocambique into the common bile duct of rats, and after 4 h of duct injection the animals were sacrificed. Concentration-response curves to acetylcholine (ACh), sodium nitroprusside (SNP) and phenylephrine (PHE) in isolated mesenteric and pulmonary arteries were obtained. Potency (pEC50) and maximal responses (EMAX) were determined. Blood samples were collected for biochemical analysis. RESULTS: In mesenteric rings, the potency for ACh was significantly decreased from animals treated with TAU (about 4.2-fold) or PLA2 (about 6.9-fold) compared to saline group without changes in the maximal responses. Neither pEC50 nor EMAX values for Ach were altered in pulmonary rings in any group. Similarly, the pEC50 and the EMAX values for SNP were not changed in both preparations in any group. The potency for PHE was significantly decreased in rat mesenteric and pulmonary rings from TAU group compared to SAL group (about 2.2- and 2.69-fold, for mesenteric and pulmonary rings, respectively). No changes were seen in the EMAX for PHE. The nitrite/nitrate (NOx-) levels were markedly increased in animals submitted to acute pancreatitis as compared to SAL group, approximately 76 and 68% in TAU and PLA2 protocol, respectively. CONCLUSION: Acute pancreatitis provoked deleterious effects in endothelium-dependent relaxing response for ACh in mesenteric rings that were strongly associated with high plasma NOx- levels as consequence of intense inflammatory responses. Furthermore, the subsensitivity of contractile response to PHE in both mesenteric and pulmonary rings might be due to the complications of this pathological condition in the early stage of pancreatitis. PMID- 18510741 TI - Leg-length inequality is not associated with greater trochanteric pain syndrome. AB - INTRODUCTION: Greater trochanteric pain syndrome (GTPS) is a common condition, the pathogenesis of which is incompletely understood. Although leg-length inequality has been suggested as a potential risk factor for GTPS, this widely held assumption has not been tested. METHODS: A cross-sectional analysis of greater trochanteric tenderness to palpation was performed in subjects with complaints of hip pain and no signs of hip osteoarthritis or generalized myofascial tenderness. Subjects were recruited from one clinical center of the Multicenter Osteoarthritis Study, a multicenter population-based study of community-dwelling adults aged 50 to 79 years. Diagnosis of GTPS was based on a standardized physical examination performed by trained examiners, and technicians measured leg length on full-limb anteroposterior radiographs. RESULTS: A total of 1,482 subjects were eligible for analysis of GTPS and leg length. Subjects' mean +/- standard deviation age was 62.4 +/- 8.2 years, and 59.8% were female. A total of 372 lower limbs from 271 subjects met the definition for having GTPS. Leg length inequality (difference > or = 1 cm) was present in 37 subjects with GTPS and in 163 subjects without GTPS (P = 0.86). Using a variety of definitions of leg-length inequality, including categorical and continuous measures, there was no association of this parameter with the occurrence of GTPS (for example, for > or = 1 cm leg-length inequality, odds ratio = 1.17 (95% confidence interval = 0.79 to 1.73)). In adjusted analyses, female sex was significantly associated with the presence of GTPS, with an adjusted odds ratio of 3.04 (95% confidence interval = 2.07 to 4.47). CONCLUSION: The present study found no evidence to support an association between leg-length inequality and greater trochanteric pain syndrome. PMID- 18510742 TI - A systematic review with procedural assessments and meta-analysis of low level laser therapy in lateral elbow tendinopathy (tennis elbow). AB - BACKGROUND: Recent reviews have indicated that low level level laser therapy (LLLT) is ineffective in lateral elbow tendinopathy (LET) without assessing validity of treatment procedures and doses or the influence of prior steroid injections. METHODS: Systematic review with meta-analysis, with primary outcome measures of pain relief and/or global improvement and subgroup analyses of methodological quality, wavelengths and treatment procedures. RESULTS: 18 randomised placebo-controlled trials (RCTs) were identified with 13 RCTs (730 patients) meeting the criteria for meta-analysis. 12 RCTs satisfied half or more of the methodological criteria. Publication bias was detected by Egger's graphical test, which showed a negative direction of bias. Ten of the trials included patients with poor prognosis caused by failed steroid injections or other treatment failures, or long symptom duration or severe baseline pain. The weighted mean difference (WMD) for pain relief was 10.2 mm [95% CI: 3.0 to 17.5] and the RR for global improvement was 1.36 [1.16 to 1.60]. Trials which targeted acupuncture points reported negative results, as did trials with wavelengths 820, 830 and 1064 nm. In a subgroup of five trials with 904 nm lasers and one trial with 632 nm wavelength where the lateral elbow tendon insertions were directly irradiated, WMD for pain relief was 17.2 mm [95% CI: 8.5 to 25.9] and 14.0 mm [95% CI: 7.4 to 20.6] respectively, while RR for global pain improvement was only reported for 904 nm at 1.53 [95% CI: 1.28 to 1.83]. LLLT doses in this subgroup ranged between 0.5 and 7.2 Joules. Secondary outcome measures of painfree grip strength, pain pressure threshold, sick leave and follow-up data from 3 to 8 weeks after the end of treatment, showed consistently significant results in favour of the same LLLT subgroup (p < 0.02). No serious side-effects were reported. CONCLUSION: LLLT administered with optimal doses of 904 nm and possibly 632 nm wavelengths directly to the lateral elbow tendon insertions, seem to offer short-term pain relief and less disability in LET, both alone and in conjunction with an exercise regimen. This finding contradicts the conclusions of previous reviews which failed to assess treatment procedures, wavelengths and optimal doses. PMID- 18510743 TI - Variable selection for large p small n regression models with incomplete data: mapping QTL with epistases. AB - BACKGROUND: Identifying quantitative trait loci (QTL) for both additive and epistatic effects raises the statistical issue of selecting variables from a large number of candidates using a small number of observations. Missing trait and/or marker values prevent one from directly applying the classical model selection criteria such as Akaike's information criterion (AIC) and Bayesian information criterion (BIC). RESULTS: We propose a two-step Bayesian variable selection method which deals with the sparse parameter space and the small sample size issues. The regression coefficient priors are flexible enough to incorporate the characteristic of "large p small n" data. Specifically, sparseness and possible asymmetry of the significant coefficients are dealt with by developing a Gibbs sampling algorithm to stochastically search through low-dimensional subspaces for significant variables. The superior performance of the approach is demonstrated via simulation study. We also applied it to real QTL mapping datasets. CONCLUSION: The two-step procedure coupled with Bayesian classification offers flexibility in modeling "large p small n" data, especially for the sparse and asymmetric parameter space. This approach can be extended to other settings characterized by high dimension and low sample size. PMID- 18510744 TI - Sequencing and genotypic analysis of the triosephosphate isomerase (TPI1) locus in a large sample of long-lived Germans. AB - BACKGROUND: Triosephosphate isomerase (TPI) is a central and conserved glycolytic enzyme. In humans, TPI is encoded by a single gene on 12p13, and associated with a rare genetic disorder, TPI deficiency. Reduced TPI activity can increase specific oxidant resistances of model organisms and TPI null-alleles have been hypothesized to promote a heterozygote advantage in man. However, comprehensive genetic information about the TPI1 locus is still lacking. RESULTS: Here, we sequenced the TPI1 locus in a sample of 357 German long-lived individuals (LLI) aged 95 to 110 years. We identified 17 different polymorphisms, of which 15 were rare and previously unknown. The two remaining SNPs occurred at much higher frequency and were tested for association with the longevity phenotype in larger samples of LLI (n = 1422) and younger controls (n = 967). Neither of the two markers showed a statistically significant difference in allele or genotype frequency between LLI and control subjects. CONCLUSION: This study marks the TPI1 locus as extraordinarily conserved, even when analyzing intronic and non-coding regions of the gene. None of the identified sequence variations affected the amino acid composition of the TPI protein and hence, are unlikely to impact the catalytic activity of the enzyme. Thus, TPI variants occur less frequent than expected and inactive alleles are not enriched in German centenarians. PMID- 18510745 TI - How high: quantity as a predictor of cannabis-related problems. AB - BACKGROUND: Research on cannabis use has emphasized frequency as a predictor of problems. Studies of other drugs reveal that frequency relates to psychological and physiological outcomes, but quantity also plays an important role. In the study of cannabis, quantity has been difficult to assess due to the wide range of products and means of consumption. METHODS: The present study introduces three new measures of quantity, and examines their contribution to cannabis-related problems. Over 5,900 adults using cannabis once or more per month completed an internet survey that inquired about use, dependence, social problems and respiratory health. In addition to detailing their frequency of cannabis use, participants also reported three measures of quantity: number of quarter ounces consumed per month, usual intensity of intoxication, and maximum intensity of intoxication. RESULTS: Frequency of use, monthly consumption, and levels of intoxication predicted respiratory symptoms, social problems and dependence. What is more, each measure of quantity accounted for significant variance in outcomes after controlling for the effects of frequency. CONCLUSION: These findings indicate that quantity is an important predictor of cannabis-related outcomes, and that the three quantity measures convey useful information about use. PMID- 18510746 TI - Variables associated with physical fighting among US high-school students. AB - BACKGROUND: Violence among adolescents is an important public health problem in the United States. This study was conducted to estimate the prevalence of having been engaged in physical fighting on school property and associated factors of the behavior among school-going adolescents in the United States. METHODS: This study was based on secondary analysis of the United States Youth Risk Behavior Survey (YRBS) conducted in 2005. The sampling frame included all private and public school in the country, stratified by region and urbanicity based on the US census bureau data. Frequencies and proportions were obtained for the outcome and explanatory variables. Logistic regression analysis was used to estimate the level of association between explanatory variables and the outcome (having been involved in a physical fight). RESULTS: Of the 13,857 respondents, 13.5% (18.2% for males and 8.8% for females) reported physical fighting onschool property in the last 12 months to the survey. Males were more likely to have been in a physical fight than females (OR = 2.23; 95% CI [1.89, 2.63]). Respondents aged 17 years or older were less likely to report physical fighting than those who were 14 years or younger. Compared to Whites, American Indians or Alaska Natives, Blacks, Native Hawaii or other Pacific Islanders, and Hispanics were more likely to report physical fighting on school property (OR = 2.11; 95% CI [1.22, 3.66], OR = 1.72; 95% CI [1.42, 2.0], OR = 2.18; 95% CI [1.01, 4.79], and OR = 1.74; 95% CI [1.41, 2.16] respectively). Physical fighting on school property was also positively associated with cigarette smoking (OR = 1.70; 95% CI [1.37, 2.10]), drinking alcohol (OR = 1.45; 95% CI [1.20, 1.76]), use of illegal drugs (OR = 1.73; 95% CI [1.42, 2.12]), having had property stolen or deliberately damaged on school property (OR = 2.06; 95% CI [1.74, 2.44]), having been threatened or injured with a weapon on school property (OR = 2.63; 95% CI [2.06, 3.34]), and playing videogame three or more hours a day (OR = 1.29; 95% CI [1.07, 1.56]). CONCLUSION: These findings suggest that physical fighting among US High School students is widespread and positively associated with victimization (having been threatened or injured with a weapon on school property) and other risky behaviors such as smoking, alcohol and drugs use. Intervention programs to prevent/control those risky behaviors as well as further attention on the association between physical fighting and victimization at school through longitudinal research are warranted. PMID- 18510747 TI - Missing value imputation for microarray gene expression data using histone acetylation information. AB - BACKGROUND: It is an important pre-processing step to accurately estimate missing values in microarray data, because complete datasets are required in numerous expression profile analysis in bioinformatics. Although several methods have been suggested, their performances are not satisfactory for datasets with high missing percentages. RESULTS: The paper explores the feasibility of doing missing value imputation with the help of gene regulatory mechanism. An imputation framework called histone acetylation information aided imputation method (HAIimpute method) is presented. It incorporates the histone acetylation information into the conventional KNN(k-nearest neighbor) and LLS(local least square) imputation algorithms for final prediction of the missing values. The experimental results indicated that the use of acetylation information can provide significant improvements in microarray imputation accuracy. The HAIimpute methods consistently improve the widely used methods such as KNN and LLS in terms of normalized root mean squared error (NRMSE). Meanwhile, the genes imputed by HAIimpute methods are more correlated with the original complete genes in terms of Pearson correlation coefficients. Furthermore, the proposed methods also outperform GOimpute, which is one of the existing related methods that use the functional similarity as the external information. CONCLUSION: We demonstrated that the using of histone acetylation information could greatly improve the performance of the imputation especially at high missing percentages. This idea can be generalized to various imputation methods to facilitate the performance. Moreover, with more knowledge accumulated on gene regulatory mechanism in addition to histone acetylation, the performance of our approach can be further improved and verified. PMID- 18510748 TI - Implementing electronic clinical reminders for lipid management in patients with ischemic heart disease in the veterans health administration: QUERI Series. AB - BACKGROUND: Ischemic heart disease (IHD) affects at least 150,000 veterans annually in the United States. Lowering serum cholesterol has been shown to reduce coronary events, cardiac death, and total mortality among high risk patients. Electronic clinical reminders available at the point of care delivery have been developed to improve lipid measurement and management in the Veterans Health Administration (VHA). Our objective was to report on a hospital-level intervention to implement and encourage use of the electronic clinical reminders. METHODS: The implementation used a quasi-experimental design with a comparison group of hospitals. In the intervention hospitals (N = 3), we used a multi faceted intervention to encourage use of the electronic clinical reminders. We evaluated the degree of reminder use and how patient-level outcomes varied at the intervention and comparison sites (N = 3), with and without adjusting for self reported reminder use. RESULTS: The national electronic clinical reminders were implemented in all of the intervention sites during the intervention period. A total of 5,438 patients with prior diagnosis of ischemic heart disease received care in the six hospitals (3 intervention and 3 comparison) throughout the 12 month intervention. The process evaluation showed variation in use of reminders at each site. Without controlling for provider self-report of use of the reminders, there appeared to be a significant improvement in lipid measurement in the intervention sites (OR 1.96, 95% CI 1.34, 2.88). Controlling for use of reminders, the amount of improvement in lipid measurement in the intervention sites was even greater (OR 2.35, CI 1.96, 2.81). Adjusting for reminder use demonstrated that only one of the intervention hospitals had a significant effect of the intervention. There was no significant change in management of hyperlipidemia associated with the intervention. CONCLUSION: There may be some benefit to focused effort to implement electronic clinical reminders, although reminders designed to improve relatively simple tasks, such as ordering tests, may be more beneficial than reminders designed to improve more complex tasks, such as initiating or titrating medications, because of the less complex nature of the task. There is value in monitoring the process, as well as outcome, of an implementation effort. PMID- 18510749 TI - The role of organizational research in implementing evidence-based practice: QUERI Series. AB - BACKGROUND: Health care organizations exert significant influence on the manner in which clinicians practice and the processes and outcomes of care that patients experience. A greater understanding of the organizational milieu into which innovations will be introduced, as well as the organizational factors that are likely to foster or hinder the adoption and use of new technologies, care arrangements and quality improvement (QI) strategies are central to the effective implementation of research into practice. Unfortunately, much implementation research seems to not recognize or adequately address the influence and importance of organizations. Using examples from the U.S. Department of Veterans Affairs (VA) Quality Enhancement Research Initiative (QUERI), we describe the role of organizational research in advancing the implementation of evidence-based practice into routine care settings. METHODS: Using the six-step QUERI process as a foundation, we present an organizational research framework designed to improve and accelerate the implementation of evidence-based practice into routine care. Specific QUERI-related organizational research applications are reviewed, with discussion of the measures and methods used to apply them. We describe these applications in the context of a continuum of organizational research activities to be conducted before, during and after implementation. RESULTS: Since QUERI's inception, various approaches to organizational research have been employed to foster progress through QUERI's six-step process. We report on how explicit integration of the evaluation of organizational factors into QUERI planning has informed the design of more effective care delivery system interventions and enabled their improved "fit" to individual VA facilities or practices. We examine the value and challenges in conducting organizational research, and briefly describe the contributions of organizational theory and environmental context to the research framework. CONCLUSION: Understanding the organizational context of delivering evidence-based practice is a critical adjunct to efforts to systematically improve quality. Given the size and diversity of VA practices, coupled with unique organizational data sources, QUERI is well-positioned to make valuable contributions to the field of implementation science. More explicit accommodation of organizational inquiry into implementation research agendas has helped QUERI researchers to better frame and extend their work as they move toward regional and national spread activities. PMID- 18510750 TI - An organizational framework and strategic implementation for system-level change to enhance research-based practice: QUERI Series. AB - BACKGROUND: The continuing gap between available evidence and current practice in health care reinforces the need for more effective solutions, in particular related to organizational context. Considerable advances have been made within the U.S. Veterans Health Administration (VA) in systematically implementing evidence into practice. These advances have been achieved through a system-level program focused on collaboration and partnerships among policy makers, clinicians, and researchers. The Quality Enhancement Research Initiative (QUERI) was created to generate research-driven initiatives that directly enhance health care quality within the VA and, simultaneously, contribute to the field of implementation science. This paradigm-shifting effort provided a natural laboratory for exploring organizational change processes. This article describes the underlying change framework and implementation strategy used to operationalize QUERI. STRATEGIC APPROACH TO ORGANIZATIONAL CHANGE: QUERI used an evidence-based organizational framework focused on three contextual elements: 1) cultural norms and values, in this case related to the role of health services researchers in evidence-based quality improvement; 2) capacity, in this case among researchers and key partners to engage in implementation research; 3) and supportive infrastructures to reinforce expectations for change and to sustain new behaviors as part of the norm. As part of a QUERI Series in Implementation Science, this article describes the framework's application in an innovative integration of health services research, policy, and clinical care delivery. CONCLUSION: QUERI's experience and success provide a case study in organizational change. It demonstrates that progress requires a strategic, systems-based effort. QUERI's evidence-based initiative involved a deliberate cultural shift, requiring ongoing commitment in multiple forms and at multiple levels. VA's commitment to QUERI came in the form of visionary leadership, targeted allocation of resources, infrastructure refinements, innovative peer review and study methods, and direct involvement of key stakeholders. Stakeholders included both those providing and managing clinical care, as well as those producing relevant evidence within the health care system. The organizational framework and related implementation interventions used to achieve contextual change resulted in engaged investigators and enhanced uptake of research knowledge. QUERI's approach and progress provide working hypotheses for others pursuing similar system-wide efforts to routinely achieve evidence-based care. PMID- 18510751 TI - Angiolymphoid hyperplasia with eosinophilia developing in a patient with history of peripheral T-cell lymphoma: evidence for multicentric T-cell lymphoproliferative process. AB - BACKGROUND: Angiolymphoid hyperplasia with eosinophilia (ALHE) is a vasocentric process characterized by infiltrates of lymphocytes and eosinophils, usually affecting the muscular arteries of the head and neck. Currently it is unclear whether it is a reactive or neoplastic process. REPORT: We present a 61-year-old African American male with a twenty year history of superficial skin patches involving the head and neck region. An excisional biopsy of a right submental lymph node revealed an atypical T-cell lymphocytic process, diagnosed as peripheral T-cell lymphoma after immunophenotyping and molecular studies. Three months later the patient underwent a biopsy of a left temporal nodule that was diagnosed as ALHE. Subsequently, at two year follow-up, the patient was diagnosed with Mycosis Fungoides. Polymerase chain reaction for T cell receptor gamma showed the same T-cell receptor gene rearrangement in both the temporal mass and the right submental lymph node. CONCLUSION: ALHE with molecular evidence of monoclonality is extremely unusual, as is the association with nodal peripheral T cell nodal lymphoma. The findings of this case support our hypothesis that ALHE might be an early form of T-cell lymphoma. PMID- 18510752 TI - Toll-like receptor 4-dependent upregulation of cytokines in a transgenic mouse model of Alzheimer's disease. AB - BACKGROUND: Abeta deposits in the brains of patients with Alzheimer's disease (AD) are closely associated with innate immune responses such as activated microglia and increased cytokines. Accumulating evidence supports the hypothesis that innate immune/inflammatory responses play a pivotal role in the pathogenesis of AD: either beneficial or harmful effects on the AD progression. The molecular mechanisms by which the innate immune system modulates the AD progression are not well understood. Toll-like receptors (TLRs) are first-line molecules for initiating the innate immune responses. When activated through TLR signaling, microglia respond to pathogens and damaged host cells by secreting chemokines and cytokines and express the co-stimulatory molecules needed for protective immune responses to pathogens and efficient clearance of damaged tissues. We previously demonstrated that an AD mouse model homozygous for a destructive mutation of TLR4 has increases in diffuse and fibrillar Abeta deposits as well as buffer-soluble and insoluble Abeta in the brain as compared with a TLR4 wild-type AD mouse model. Here, we investigated the roles of TLR4 in Abeta-induced upregulation of cytokines and chemokines, Abeta-induced activation of microglia and astrocytes and Abeta-induced immigration of leukocytes. METHODS: Using the same model, levels of cytokines and chemokines in the brain were determined by multiplex cytokine/chemokine array. Activation of microglia and astrocytes and immigration of leukocytes were determined by immunoblotting and immunohistochemistry followed by densitometry and morphometry, respectively. RESULTS: Levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-10 and IL-17 in the brains of TLR4 wild-type AD mice were significantly higher than those in TLR4 wild-type non transgenic littermates. Such increases in cytokines were not found in TLR4 mutant AD mice as compared with TLR4 mutant non-transgenic littermates. Although expression levels of CD11b (a microglia marker) and GFAP (a reactive astrocyte marker) in the brains of TLR4 mutant AD mice were higher than those in TLR4 wild type AD mice, no difference was found in levels of CD45 (common leukocyte antigen). CONCLUSION: This is the first demonstration of TLR4-dependent upregulation of cytokines in an AD mouse model. Our results suggest that TLR4 signaling is involved in AD progression and that TLR4 signaling can be a new therapeutic target for AD. PMID- 18510753 TI - Providing support to psychiatric patients living in the community in Japan: patient needs and care providers perceptions. AB - BACKGROUND: Social support programs are a critical component of care for psychiatric patients living in community or residential settings. There is little information, however, on how to optimally deliver these services in the Japanese context. METHODS: We selected ten community life support centers for patients with major mental illness and administered questionnaires to 199 pairs of patients and staff members. These questionnaires consisted of twenty-six items from six categories: difficulties with interpersonal relationships; risks to physical well-being; risks to mental health; difficulties with life skills; challenges regarding living conditions; risks towards community safety. For each of these items, patients were asked whether they had experienced difficulties during the previous month, and staff members were asked the extent to which their patients needed support. RESULTS: The results demonstrated that staff members tended to understate patients' needs regarding chronic medical conditions (p < 0.01), dietary habits (< 0.01), and excessive smoking or alcohol drinking (< 0.05). On the other hand, staff members recognized patients' needs regarding mental health problems to a greater extent than patients themselves (< 0.05). CONCLUSION: Results of this study suggest that social services geared towards specific tasks of daily living form an important component of comprehensive care for psychiatric patients living in community settings in Japan. PMID- 18510754 TI - His-tag ELISA for the detection of humoral tumor-specific immunity. AB - BACKGROUND: The application of high throughput molecular techniques such as SEREX are resulting in the identification of a multitude of tumor associated antigens. As newly identified antigens are incorporated into a variety of clinical trials, standardization of immunologic monitoring methods becomes increasingly important. We questioned whether mammalian cell expression of a histadine-linked human protein could be used to produce antigen suitable for detecting tumor-specific humoral immunity and whether such an assay could be amenable to standardization for clinical use. METHODS: We designed a his-tagged capture ELISA based on lysate from genetically engineered CHO cells for detection of antibodies to insulin-like growth factor binding protein 2, a novel tumor antigen. We performed technical and preliminary clinical validation studies, including comparison to a standard indirect ELISA based on commercially prepared recombinant antigen. RESULTS: The his-tagged capture ELISA could be standardized. Precision experiments resulted in CVs < 15%. Linearity and calibration experiments demonstrated r2 values of 0.99. In comparison to Western blot analysis, his-tag and indirect ELISA accurately identified 88% and 93% of samples, respectively. Sample concordance between capture and indirect assays was highly significant (p = 0.003). Furthermore, significantly greater levels of IGFBP-2 antibody immunity were found in cancer patients compared to normal controls (p = 0.008). CONCLUSION: A genetically engineered cell lysate based ELISA can be amenable to standardization and can detect increased levels of antibody immunity to tumor-associated antigen in cancer patients compared to non tumor-bearing healthy controls. PMID- 18510755 TI - Genomic organization of zebrafish microRNAs. AB - BACKGROUND: microRNAs (miRNAs) are small (~22 nt) non-coding RNAs that regulate cell movement, specification, and development. Expression of miRNAs is highly regulated, both spatially and temporally. Based on direct cloning, sequence conservation, and predicted secondary structures, a large number of miRNAs have been identified in higher eukaryotic genomes but whether these RNAs are simply a subset of a much larger number of noncoding RNA families is unknown. This is especially true in zebrafish where genome sequencing and annotation is not yet complete. RESULTS: We analyzed the zebrafish genome to identify the number and location of proven and predicted miRNAs resulting in the identification of 35 new miRNAs. We then grouped all 415 zebrafish miRNAs into families based on seed sequence identity as a means to identify possible functional redundancy. Based on genomic location and expression analysis, we also identified those miRNAs that are likely to be encoded as part of polycistronic transcripts. Lastly, as a resource, we compiled existing zebrafish miRNA expression data and, where possible, listed all experimentally proven mRNA targets. CONCLUSION: Current analysis indicates the zebrafish genome encodes 415 miRNAs which can be grouped into 44 families. The largest of these families (the miR-430 family) contains 72 members largely clustered in two main locations along chromosome 4. Thus far, most zebrafish miRNAs exhibit tissue specific patterns of expression. PMID- 18510756 TI - A new approach to understanding the impact of circadian disruption on human health. AB - BACKGROUND: Light and dark patterns are the major synchronizer of circadian rhythms to the 24-hour solar day. Disruption of circadian rhythms has been associated with a variety of maladies. Ecological studies of human exposures to light are virtually nonexistent, however, making it difficult to determine if, in fact, light-induced circadian disruption directly affects human health. METHODS: A newly developed field measurement device recorded circadian light exposures and activity from day-shift and rotating-shift nurses. Circadian disruption defined in terms of behavioral entrainment was quantified for these two groups using phasor analyses of the circular cross-correlations between light exposure and activity. Circadian disruption also was determined for rats subjected to a consistent 12-hour light/12-hour dark pattern (12L:12D) and ones subjected to a "jet-lagged" schedule. RESULTS: Day-shift nurses and rats exposed to the consistent light-dark pattern exhibited pronounced similarities in their circular cross-correlation functions and 24-hour phasor representations except for an approximate 12-hour phase difference between species. The phase difference reflects the diurnal versus nocturnal behavior of humans versus rodents. Phase differences within species likely reflect chronotype differences among individuals. Rotating-shift nurses and rats subjected to the "jet-lagged" schedule exhibited significant reductions in phasor magnitudes compared to the day-shift nurses and the 12L:12D rats. The reductions in the 24-hour phasor magnitudes indicate a loss of behavioral entrainment compared to the nurses and the rats with regular light-dark exposure patterns. CONCLUSION: This paper provides a quantitative foundation for systematically studying the impact of light-induced circadian disruption in humans and in animal models. Ecological light and activity data are needed to develop the essential insights into circadian entrainment/disruption actually experienced by modern people. These data can now be obtained and analyzed to reveal the interrelationship between actual light exposures and markers of circadian rhythm such as rest-activity patterns, core body temperature, and melatonin synthesis. Moreover, it should now be possible to bridge ecological studies of circadian disruption in humans to parametric studies of the relationships between circadian disruption and health outcomes using animal models. PMID- 18510757 TI - Executive function does not predict coping with symptoms in stable patients with a diagnosis of schizophrenia. AB - BACKGROUND: Associations between coping with and control over psychotic symptoms were examined using the Maastricht Assessment of Coping Strategies-24, testing the hypothesis that the cognitive domain of executive functioning predicted quality and quantity of coping. METHODS: MACS-24 was administered to 32 individuals with a diagnosis of schizophrenia. For each of 24 symptoms, experience of distress, type of coping and the resulting degree of perceived control were assessed. Coping types were reduced to two contrasting coping categories: symptomatic coping (SC) and non-symptomatic coping (NSC; combining active problem solving, passive illness behaviour, active problem avoiding, and passive problem avoiding). Cognitive functioning was assessed using the GIT (Groninger Intelligence Test), the Zoo map (BADS: Behavioural Assessment of Dysexecutive function), Stroop-test and Trail making. RESULTS: Cognitive function was not associated with frequency of coping, nor did cognitive function differentially predict SC or NSC. Cognitive function similarly was not associated with symptom distress or level of perceived control over the symptom. CONCLUSION: There was no evidence that cognitive function predicts quantity or quality of coping with symptoms in people with a diagnosis of schizophrenia. Variation in the realm of emotion regulation and social cognition may be more predictive of coping with psychotic symptoms. PMID- 18510758 TI - Exploiting orthologue diversity for systematic detection of gain-of-function phenotypes. AB - BACKGROUND: Systematic search for genes whose gain-of-function by exogenous expression confers an advantage in cell-based selective screenings is a powerful method for unbiased functional exploration of the genome, and has the potential to disclose new targets for cancer therapy. A major limit of this approach resides in the labor-intensive cloning of resistant cells, identification of the integrated genes and validation of their ability to confer a selective advantage. Moreover, the selection has to be drastic and genes conferring a limited advantage are typically missed. RESULTS: We developed a new functional screening strategy based on transduction of mammalian cells of a given species with an expression library from another species, followed by one-shot quantitative tracing with DNA microarrays of all library-derived transcripts before and after selection. In this way, exogenous transcripts enriched after selection, and therefore likely to confer resistance, are readily detected. We transduced a retroviral cDNA expression library from mouse testis into human and canine cells, and optimized the use of commercial murine gene expression arrays for species specific detection of library-derived transcripts. We then conducted a functional screening by growing library-transduced canine MDCK cells in suspension, to enrich for cDNAs conferring anchorage independence. Notably, these cells show partial resistance to loss of anchorage, and the selection can be of limited stringency, compromising approaches based on clonal selection or anyway requiring high stringency. Microarray analysis revealed reproducible enrichment after three weeks of growth on polyhema for seven genes, among which the Hras proto-oncogene and Sox5. When individually transduced into MDCK cells, Sox5 specifically promoted anchorage-independent growth, thereby confirming the validity and specificity of the approach. CONCLUSION: The procedure described here brings substantial advantages to the field of expression cloning, being faster, more systematic and more sensitive. Indeed, this strategy allowed identification and validation of genes promoting anchorage-independent growth of epithelial cells under selection conditions not amenable to conventional expression cloning. PMID- 18510759 TI - Extended flow cytometry characterization of normal bone marrow progenitor cells by simultaneous detection of aldehyde dehydrogenase and early hematopoietic antigens: implication for erythroid differentiation studies. AB - BACKGROUND: Aldehyde dehydrogenase (ALDH) is a cytosolic enzyme highly expressed in hematopoietic precursors from cord blood and granulocyte-colony stimulating factor mobilized peripheral blood, as well as in bone marrow from patients with acute myeloblastic leukemia. As regards human normal bone marrow, detailed characterization of ALDH+ cells has been addressed by one single study (Gentry et al, 2007). The goal of our work was to provide new information about the dissection of normal bone marrow progenitor cells based upon the simultaneous detection by flow cytometry of ALDH and early hematopoietic antigens, with particular attention to the expression of ALDH on erythroid precursors. To this aim, we used three kinds of approach: i) multidimensional analytical flow cytometry, detecting ALDH and early hematopoietic antigens in normal bone marrow; ii) fluorescence activated cell sorting of distinct subpopulations of progenitor cells, followed by in vitro induction of erythroid differentiation; iii) detection of ALDH+ cellular subsets in bone marrow from pure red cell aplasia patients. RESULTS: In normal bone marrow, we identified three populations of cells, namely ALDH+CD34+, ALDH-CD34+ and ALDH+CD34- (median percentages were 0.52, 0.53 and 0.57, respectively). As compared to ALDH-CD34+ cells, ALDH+CD34+ cells expressed the phenotypic profile of primitive hematopoietic progenitor cells, with brighter expression of CD117 and CD133, accompanied by lower display of CD38 and CD45RA. Of interest, ALDH+CD34- population disclosed a straightforward erythroid commitment, on the basis of three orders of evidences. First of all, ALDH+CD34- cells showed a CD71bright, CD105+, CD45- phenotype. Secondly, induction of differentiation experiments evidenced a clear-cut expression of glycophorin A (CD235a). Finally, ALDH+CD34- precursors were not detectable in patients with pure red cell aplasia (PRCA). CONCLUSION: Our study, comparing surface antigen expression of ALDH+/CD34+, ALDH-/CD34+ and ALDH+/CD34- progenitor cell subsets in human bone marrow, clearly indicated that ALDH+CD34- cells are mainly committed towards erythropoiesis. To the best of our knowledge this finding is new and could be useful for basic studies about normal erythropoietic differentiation as well as for enabling the employment of ALDH as a red cell marker in polychromatic flow cytometry characterization of bone marrow from patients with aplastic anemia and myelodysplasia. PMID- 18510760 TI - BatchPrimer3: a high throughput web application for PCR and sequencing primer design. AB - BACKGROUND: Microsatellite (simple sequence repeat - SSR) and single nucleotide polymorphism (SNP) markers are two types of important genetic markers useful in genetic mapping and genotyping. Often, large-scale genomic research projects require high-throughput computer-assisted primer design. Numerous such web-based or standard-alone programs for PCR primer design are available but vary in quality and functionality. In particular, most programs lack batch primer design capability. Such a high-throughput software tool for designing SSR flanking primers and SNP genotyping primers is increasingly demanded. RESULTS: A new web primer design program, BatchPrimer3, is developed based on Primer3. BatchPrimer3 adopted the Primer3 core program as a major primer design engine to choose the best primer pairs. A new score-based primer picking module is incorporated into BatchPrimer3 and used to pick position-restricted primers. BatchPrimer3 v1.0 implements several types of primer designs including generic primers, SSR primers together with SSR detection, and SNP genotyping primers (including single-base extension primers, allele-specific primers, and tetra-primers for tetra-primer ARMS PCR), as well as DNA sequencing primers. DNA sequences in FASTA format can be batch read into the program. The basic information of input sequences, as a reference of parameter setting of primer design, can be obtained by pre-analysis of sequences. The input sequences can be pre-processed and masked to exclude and/or include specific regions, or set targets for different primer design purposes as in Primer3Web and primer3Plus. A tab-delimited or Excel-formatted primer output also greatly facilitates the subsequent primer-ordering process. Thousands of primers, including wheat conserved intron-flanking primers, wheat genome-specific SNP genotyping primers, and Brachypodium SSR flanking primers in several genome projects have been designed using the program and validated in several laboratories. CONCLUSION: BatchPrimer3 is a comprehensive web primer design program to develop different types of primers in a high-throughput manner. Additional methods of primer design can be easily integrated into future versions of BatchPrimer3. The program with source code and thousands of PCR and sequencing primers designed for wheat and Brachypodium are accessible at http://wheat.pw.usda.gov/demos/BatchPrimer3/. PMID- 18510761 TI - Genome-wide gene expression profiling analysis of Leishmania major and Leishmania infantum developmental stages reveals substantial differences between the two species. AB - BACKGROUND: Leishmania parasites cause a diverse spectrum of diseases in humans ranging from spontaneously healing skin lesions (e.g., L. major) to life threatening visceral diseases (e.g., L. infantum). The high conservation in gene content and genome organization between Leishmania major and Leishmania infantum contrasts their distinct pathophysiologies, suggesting that highly regulated hierarchical and temporal changes in gene expression may be involved. RESULTS: We used a multispecies DNA oligonucleotide microarray to compare whole-genome expression patterns of promastigote (sandfly vector) and amastigote (mammalian macrophages) developmental stages between L. major and L. infantum. Seven per cent of the total L. infantum genome and 9.3% of the L. major genome were differentially expressed at the RNA level throughout development. The main variations were found in genes involved in metabolism, cellular organization and biogenesis, transport and genes encoding unknown function. Remarkably, this comparative global interspecies analysis demonstrated that only 10-12% of the differentially expressed genes were common to L. major and L. infantum. Differentially expressed genes are randomly distributed across chromosomes further supporting a posttranscriptional control, which is likely to involve a variety of 3'UTR elements. CONCLUSION: This study highlighted substantial differences in gene expression patterns between L. major and L. infantum. These important species-specific differences in stage-regulated gene expression may contribute to the disease tropism that distinguishes L. major from L. infantum. PMID- 18510762 TI - Protocol for stage 2 of the GaP study (genetic testing acceptability for Paget's disease of bone): a questionnaire study to investigate whether relatives of people with Paget's disease would accept genetic testing and preventive treatment if they were available. AB - BACKGROUND: Paget's disease of bone (PDB) disrupts normal bone architecture and causes pain, deformity, deafness, osteoarthritis, and fractures. Genetic factors play a role in PDB and genetic tests are now conducted for research purposes. It is thus timely to investigate the potential for a clinical programme of genetic testing and preventative treatment for people who have a family history of PDB. This study examines the beliefs of relatives of people with PDB. It focuses particularly on illness and treatment representations as predictors of the acceptability and uptake of potential clinical programmes. Illness representations are examined using Leventhal's Common Sense Self-Regulation Model while cognitions about treatment behaviours (acceptance of testing and treatment uptake) are conceptualised within the Theory of Planned Behaviour. METHODS/DESIGN: A postal questionnaire of non-affected relatives of people with Paget's disease. The sample will include relatives of Paget's patients with a family history of Paget's disease and relatives of Paget's patients without a family history of Paget's disease. The questionnaire will explore whether a range of factors relate to acceptability of a programme of genetic testing and preventive treatment in relatives of Paget's disease sufferers. The questionnaire will include several measures: illness representations (as measured by the Brief Illness Perceptions Questionnaire); treatment representations (as measured by Theory of Planned Behaviour-based question items, informed by a prior interview elicitation study); descriptive and demographic details; and questions exploring family environment and beliefs of other important people. Data will also be collected from family members who have been diagnosed with Paget's disease to describe the disease presentation and its distribution within a family. DISCUSSION: The answers to these measures will inform the feasibility of a programme of genetic testing and preventive treatment for individuals who are at a high risk of developing Paget's disease because they carry an appropriate genetic mutation. They will also contribute to theoretical and empirical approaches to predicting diagnostic and treatment behaviours from the combined theoretical models. PMID- 18510763 TI - Vertebral osteomyelitis and native valve endocarditis due to Staphylococcus simulans: a case report. AB - BACKGROUND: Staphylococcus simulans is a common animal pathogen that occasionally can colonize human skin. Unlike other coagulase-negative staphylococci, S. simulans tends to cause more severe infections that resemble those caused by S. aureus. We present a case of vertebral osteomyelitis and endocarditis due to S. simulans. To the best of our knowledge, this is the first report of vertebral osteomyelitis associated with native valve endocarditis rather than orthopedic surgery. CASE PRESENTATION: A 46-year-old male butcher was admitted to the hospital with a 4-week history of high fever with profound sweating. He reported weakness in his legs and low back pain that compromised his walking ability. Blood cultures yielded Gram-positive cocci on Gram stain. These cocci were identified to the species level as S. simulans, a coagulase-negative staphylococcus. The patient was treated with antibiotics, which were discontinued after 6 months. CONCLUSION: This case illustrates the importance of identifying coagulase-negative staphylococci to the species level. Accurate identification of S. simulans would further help investigations defining its pathogenic role in human infections. PMID- 18510764 TI - Development and optimization of quantitative PCR for the diagnosis of invasive aspergillosis with bronchoalveolar lavage fluid. AB - BACKGROUND: The diagnosis of invasive pulmonary aspergillosis (IPA) remains challenging. Culture and histopathological examination of bronchoalveolar lavage (BAL) fluid are useful but have suboptimal sensitivity and in the case of culture may require several days for fungal growth to be evident. Detection of Aspergillus DNA in BAL fluid by quantitative PCR (qPCR) offers the potential for earlier diagnosis and higher sensitivity. It is important to adopt quality control measures in PCR assays to address false positives and negatives which can hinder accurate evaluation of diagnostic performance. METHODS: BAL fluid from 94 episodes of pneumonia in 81 patients was analyzed. Thirteen episodes were categorized as proven or probable IPA using Mycoses Study Group criteria. The pellet and the supernatant fractions of the BAL were separately assayed. A successful extraction was confirmed with a human 18S rRNA gene qPCR. Inhibition in each qPCR was measured using an exogenous DNA based internal amplification control (IAC). The presence of DNA from pathogens in the Aspergillus genus was detected using qPCR targeting fungal 18S rRNA gene. RESULTS: Human 18S rRNA gene qPCR confirmed successful DNA extraction of all samples. IAC detected some degree of initial inhibition in 11 samples. When culture was used to diagnose IPA, the sensitivity and specificity were 84.5% and 100% respectively. Receiver-operating characteristic analysis of qPCR showed that a cutoff of 13 fg of Aspergillus genomic DNA generated a sensitivity, specificity, positive and negative predictive value of 77%, 88%, 50%, 96% respectively. BAL pellet and supernatant analyzed together resulted in sensitivity and specificity similar to BAL pellet alone. Some patients did not meet standard criteria for IPA, but had consistently high levels of Aspergillus DNA in BAL fluid by qPCR. CONCLUSION: The Aspergillus qPCR assay detected Aspergillus DNA in 76.9% of subjects with proven or probable IPA when the concentrated BAL fluid pellet fraction was used for diagnosis. There was no benefit from analyzing the BAL supernatant fraction. Use of both extraction and amplification controls provided optimal quality control for interpreting qPCR results and therefore may increase our understanding of the true potential of qPCR for the diagnosis of IPA. PMID- 18510765 TI - Internet image viewer (iiV). AB - BACKGROUND: Visualizing 3-dimensional (3-D) datasets is an important part of modern neuroimaging research. Many tools address this problem; however, they often fail to address specific needs and flexibility, such as the ability to work with different data formats, to control how and what data are displayed, to interact with values, and to undo mistakes. RESULTS: iiV, an interactive software program for displaying 3-D brain images, is described. This tool was programmed to solve basic problems in 3-D data visualization. It is written in Java so it is extensible, is platform independent, and can display images within web pages.iiV displays 3-D images as 2-dimensional (2-D) slices with each slice being an independent object with independent features such as location, zoom, colors, labels, etc. Feature manipulation becomes easier by having a full set of editing capabilities including the following: undo or redo changes; drag, copy, delete and paste objects; and save objects with their features to a file for future editing. It can read multiple standard positron emission tomography (PET) and magnetic resonance imaging (MRI) file formats like ECAT, ECAT7, ANALYZE, NIfTI-1 and DICOM. We present sample applications to illustrate some of the features and capabilities. CONCLUSION: iiV is an image display tool with many useful features. It is highly extensible, platform independent, and web-compatible. This report summarizes its features and applications, while illustrating iiV's usefulness to the biomedical imaging community. PMID- 18510766 TI - Nef does not contribute to replication differences between R5 pre-AIDS and AIDS HIV-1 clones from patient ACH142. AB - AIDS-associated, CCR5-tropic (R5) HIV-1 clones, isolated from a patient that never developed CXCR4-tropic HIV-1, replicate to a greater extent and cause greater cytopathic effects than R5 HIV-1 clones isolated before the onset of AIDS. Previously, we showed that HIV-1 Env substantially contributed to the enhanced replication of an AIDS clone. In order to determine if Nef makes a similar contribution, we cloned and phenotypically analyzed nef genes from a series of patient ACH142 derived R5 HIV-1 clones. The AIDS-associated Nef contains a series of residues found in Nef proteins from progressors 1. In contrast to other reports 123, this AIDS-associated Nef downmodulated MHC-I to a greater extent and CD4 less than pre-AIDS Nef proteins. Additionally, all Nef proteins enhanced infectivity similarly in a single round of replication. Combined with our previous study, these data show that evolution of the HIV-1 env gene, but not the nef gene, within patient ACH142 significantly contributed to the enhanced replication and cytopathic effects of the AIDS-associated R5 HIV-1 clone. PMID- 18510767 TI - Sleep disturbances in an arctic population: the Tromso Study. AB - BACKGROUND: Prevalence estimates for insomnia range from 10 to 50% in the adult general population. Sleep disturbances cause great impairment in quality of life, which might even rival or exceed the impairment in other chronic medical disorders. The economic implications and use of health-care services related to chronic insomnia represent a clinical concern as well as a pronounced public health problem. Hypnotics are frequently prescribed for insomnia, but alcohol and over-the-counter sleep aids seem to be more widely used by insomniacs than prescription medications. Despite the complex relationship between insomnia and physical and mental health factors, the condition appears to be underrecognized and undertreated by health care providers, probably due to the generally limited knowledge of the causes and natural development of insomnia. METHODS/DESIGN: The Tromso Study is an ongoing population-based cohort study with five previous health studies undertaken between 1974 and 2001. This protocol outlines a planned study within the sixth Tromso Study (Tromso VI), aiming at; 1) describing sleep patterns in a community-based sample representative of the general population of northern Norway, and 2) examining outcome variables of sleep disturbances against possible explanatory and confounding variables, both within a cross-sectional approach, as well as retrospectively in a longitudinal study - exploring sleep patterns in subjects who have attended two or more of the previous Tromso studies between 1974 and 2009. First, we plan to perform a simple screening in order to identify those participants with probable sleep disturbances, and secondly to investigate these sleep disturbances further, using an extensive sleep questionnaire. We will also collect biological explanatory variables, i.e. blood samples, weight, height and blood pressure. We plan to merge data on an individual level from the Tromso VI Study with data from the Norwegian Prescription Database (NorPD), which is a national registry including data for all prescription drugs issued at Norwegian pharmacies. Participants with sleep disturbances will be compared with pair-matched controls without sleep disturbances. DISCUSSION: Despite ongoing research, many challenges remain in the characterization of sleep disturbances and its correlates. Future mapping of the biological dimensions, natural history, as well as the behavioral and drug related aspects of sleep disturbances in a representative population samples is clearly needed. PMID- 18510768 TI - Evolution of the CDKN1C-KCNQ1 imprinted domain. AB - BACKGROUND: Genomic imprinting occurs in both marsupial and eutherian mammals. The CDKN1C and IGF2 genes are both imprinted and syntenic in the mouse and human, but in marsupials only IGF2 is imprinted. This study examines the evolution of features that, in eutherians, regulate CDKN1C imprinting. RESULTS: Despite the absence of imprinting, CDKN1C protein was present in the tammar wallaby placenta. Genomic analysis of the tammar region confirmed that CDKN1C is syntenic with IGF2. However, there are fewer LTR and DNA elements in the region and in intron 9 of KCNQ1. In addition there are fewer LINEs in the tammar compared with human and mouse. While the CpG island in intron 10 of KCNQ1 and promoter elements could not be detected, the antisense transcript KCNQ1OT1 that regulates CDKN1C imprinting in human and mouse is still expressed. CONCLUSION: CDKN1C has a conserved function, likely antagonistic to IGF2, in the mammalian placenta that preceded its acquisition of imprinting. CDKN1C resides in synteny with IGF2, demonstrating that imprinting of the two genes did not occur concurrently to balance maternal and paternal influences on the growth of the placenta. The expression of KCNQ1OT1 in the absence of CDKN1C imprinting suggests that antisense transcription at this locus preceded imprinting of this domain. These findings demonstrate the stepwise accumulation of control mechanisms within imprinted domains and show that CDKN1C imprinting cannot be due to its synteny with IGF2 or with its placental expression in mammals. PMID- 18510769 TI - Human papillomavirus (HPV) infection and intraepithelial neoplasia and invasive cancer of the uterine cervix: a case-control study in Zaragoza, Spain. AB - INTRODUCTION: The raw incidence of cancer of the uterine cervix is Spain is 7,8 per 100.000 inhabitants (adjusted incidence is 5.6). The incidence of this tumor is still low, but a steady increase has been seen, probably related to increasing risk factors. AIM: To determine the frequency of infection by different types of human papillomavirus (HPV) in Papanicolau smears from women with and without cancer of the uterine cervix in Spain. PATIENTS AND METHODS: A case-control study was performed in women with and without cervical cancer from Zaragoza, Spain. Pap smears from 600 cases (540 women with cervical intraepithelial neoplasms (CIN) and 60 with invasive cancer) and 1200 controls (women without those lesions) were tested by polymerase chain reaction (PCR) and typed by oligonucleotide microarray based detection. RESULTS: HPV was detected in 93.3% of all samples with invasive cancer versus 17.5% of controls. OR for invasive cancer was 55 (95% CI 21.5 140,5). Statistically significant associations were also found for different grades of cervical dysplasia. CONCLUSION: The strong association found between HPV infection, specifically types 16 and 18 and cancer of the uterine cervix in Zaragoza, Spain, stresses the importance of ongoing efforts to institute a vaccine program with recently approved HPV vaccines in order to prevent cervical cancer in this population. PMID- 18510770 TI - Psychopathological status, behavior problems, and family adjustment of Kuwaiti children whose fathers were involved in the first gulf war. AB - OBJECTIVES: Following the end of the Gulf War that resulted in the liberation of Kuwait, there are no reports on the impact of veterans' traumatic exposure and posttraumatic stress disorder (PTSD) on their children. We compared the severity of anxiety, depression, deviant behavior and poor family adjustment among the children of a stratified random sample of four groups of Kuwaiti military men, viz: the retired; an active -in-the-army group (AIA) (involved in duties at the rear); an in-battle group (IB) (involved in combat); and a prisoners -of- war (POWs) group. Also, we assessed the association of father's PTSD/combat status and mother's characteristics with child psychosocial outcomes. METHOD: Subjects were interviewed at home, 6 years after the war, using: the Child Behavior Index to assess anxiety, depression, and adaptive behavior; Rutter Scale A2 for deviant behavior; and Family Adjustment Device for adjustment at home. Both parents were assessed for PTSD. RESULTS: The 489 offspring (250 m, 239 f; mean age 13.8 yrs) belonged to 166 father-mother pairs. Children of POWs tended to have higher anxiety, depression, and abnormal behavior scores. Those whose fathers had PTSD had significantly higher depression scores. However, children of fathers with both PTSD and POW status (N = 43) did not have significantly different outcome scores than the other father PTSD/combat status groups. Mother's PTSD, anxiety, depression and social status were significantly associated with all the child outcome variables. Parental age, child's age and child's level of education were significant covariates. Although children with both parents having PTSD had significantly higher anxiety/depression scores, the mother's anxiety was the most frequent and important predictor of child outcome variables. The frequency of abnormal test scores was: 14% for anxiety/depression, and 17% for deviant behavior. CONCLUSION: Our findings support the impression that child emotional experiences in vulnerable family situations transcend culture and are associated with the particular behavior of significant adults in the child's life. The primacy of the mother's influence has implications for interventions to improve the psychological functioning of children in such families. Mental health education for these families has the potential to help those in difficulty. PMID- 18510771 TI - Development and evaluation of a high-throughput, low-cost genotyping platform based on oligonucleotide microarrays in rice. AB - BACKGROUND: We report the development of a microarray platform for rapid and cost effective genetic mapping, and its evaluation using rice as a model. In contrast to methods employing whole-genome tiling microarrays for genotyping, our method is based on low-cost spotted microarray production, focusing only on known polymorphic features. RESULTS: We have produced a genotyping microarray for rice, comprising 880 single feature polymorphism (SFP) elements derived from insertions/deletions identified by aligning genomic sequences of the japonica cultivar Nipponbare and the indica cultivar 93-11. The SFPs were experimentally verified by hybridization with labeled genomic DNA prepared from the two cultivars. Using the genotyping microarrays, we found high levels of polymorphism across diverse rice accessions, and were able to classify all five subpopulations of rice with high bootstrap support. The microarrays were used for mapping of a gene conferring resistance to Magnaporthe grisea, the causative organism of rice blast disease, by quantitative genotyping of samples from a recombinant inbred line population pooled by phenotype. CONCLUSION: We anticipate this microarray based genotyping platform, based on its low cost-per-sample, to be particularly useful in applications requiring whole-genome molecular marker coverage across large numbers of individuals. PMID- 18510772 TI - Pyrosequencing as a method for SNP identification in the rhesus macaque (Macaca mulatta). AB - BACKGROUND: Rhesus macaques (Macaca mulatta) are the primate most used for biomedical research, but phenotypic differences between Indian-origin and Chinese rhesus macaques have encouraged genetic methods for identifying genetic differences between these two populations. The completion of the rhesus genome has led to the identification of many single nucleotide polymorphisms (SNPs) in this species. These single nucleotide polymorphisms have many advantages over the short tandem repeat (STR) loci currently used to assay genetic variation. However, the number of currently identified polymorphisms is too small for whole genome analysis or studies of quantitative trait loci. To that end, we tested a combination of methods to identify large numbers of high-confidence SNPs, and screen those with high minor allele frequencies (MAF). RESULTS: By testing our previously reported single nucleotide polymorphisms, we identified a subset of high-confidence, high-MAF polymorphisms. Resequencing revealed a large number of regionally specific SNPs not identified through a single pyrosequencing run. By resequencing a pooled sample of four individuals, we reliably identified loci with a MAF of at least 12.5%. Finally, we found that when applied to a larger, geographically variable sample of rhesus, a large proportion of our loci were variable in both populations, and very few loci were ancestry informative. Despite this fact, the SNP loci were more effective at discriminating Indian and Chinese rhesus than STR loci. CONCLUSION: Pyrosequencing and pooled resequencing are viable methods for the identification of high-MAF SNP loci in rhesus macaques. These SNP loci are appropriate for screening both the inter- and intra population genetic variation. PMID- 18510773 TI - Short-term triple therapy with azithromycin for Helicobacter pylori eradication: low cost, high compliance, but low efficacy. AB - BACKGROUND: The Brazilian consensus recommends a short-term treatment course with clarithromycin, amoxicillin and proton-pump inhibitor for the eradication of Helicobacter pylori (H. pylori). This treatment course has good efficacy, but cannot be afforded by a large part of the population. Azithromycin, amoxicillin and omeprazole are subsidized, for several aims, by the Brazilian federal government. Therefore, a short-term treatment course that uses these drugs is a low-cost one, but its efficacy regarding the bacterium eradication is yet to be demonstrated. The study's purpose was to verify the efficacy of H. pylori eradication in infected patients who presented peptic ulcer disease, using the association of azithromycin, amoxicillin and omeprazole. METHODS: Sixty patients with peptic ulcer diagnosed by upper digestive endoscopy and H. pylori infection documented by rapid urease test, histological analysis and urea breath test were treated for six days with a combination of azithromycin 500 mg and omeprazole 20 mg, in a single daily dose, associated with amoxicillin 500 mg 3 times a day. The eradication control was carried out 12 weeks after the treatment by means of the same diagnostic tests. The eradication rates were calculated with 95% confidence interval. RESULTS: The eradication rate was 38% per intention to treat and 41% per protocol. Few adverse effects were observed and treatment compliance was high. CONCLUSION: Despite its low cost and high compliance, the low eradication rate does not allow the recommendation of the triple therapy with azithromycin as an adequate treatment for H. pylori infection. PMID- 18510774 TI - Identifying outcome-based indicators and developing a curriculum for a continuing medical education programme on rational prescribing using a modified Delphi process. AB - BACKGROUND: Continuing medical education (CME) is compulsory for physicians in Iran. Recent studies in Iran show that modifications of CME elements are necessary to improve the effectiveness of the educational programmes. Other studies point to an inappropriate, even irrational drug prescribing. Based on a needs assessment study regarding CME for general physicians in the East Azerbaijan province in Iran, rational prescribing practice was recognized as a high priority issue. Considering different educational methods, outcome-based education has been proposed as a suitable approach for CME. The purpose of the study was to obtain experts' consensus about appropriate educational outcomes of rational prescribing for general physicians in CME and developing curricular contents for this education. METHODS: The study consisted of two phases: The first phase was conducted using a two-round Delphi consensus process to identify the outcome-based educational indicators regarding rational prescribing for general physicians in primary care (GPs). In the second phase the agreed indicators were submitted to panels of experts for assessment and determination of content for a CME program in the field. RESULTS: Twenty one learning outcomes were identified through a modified Delphi process. The indicators were used by the panels of experts and six educational topics were determined for the CME programme and the curricular content of each was defined. The topics were 1) Principles of prescription writing, 2) Adverse drug reactions, 3) Drug interactions, 4) Injections, 5) Antibiotic therapy, and 6) Anti-inflammatory agents therapy. One of the topics was not directly related to any outcome, raising a question about the need for a discussion on constructive alignment. CONCLUSIONS: Consensus on learning outcomes was achieved and an educational guideline was designed. Before suggesting widespread use in the country the educational package should be tested in the CME context. PMID- 18510775 TI - The complete mitochondrial genome sequence of the hydrothermal vent galatheid crab Shinkaia crosnieri (Crustacea: Decapoda: Anomura): a novel arrangement and incomplete tRNA suite. AB - BACKGROUND: Metazoan mitochondrial genomes usually consist of the same 37 genes. Such genes contain useful information for phylogenetic analyses and evolution modelling. Although complete mitochondrial genomes have been determined for over 1,000 animals to date, hydrothermal vent species have, thus far, remained excluded due to the scarcity of collected specimens. RESULTS: The mitochondrial genome of the hydrothermal vent galatheid crab Shinkaia crosnieri is 15,182 bp in length, and is composed of 13 protein-coding genes, two ribosomal RNA genes and only 18 transfer RNA genes. The total AT content of the genome, as is typical for decapods, is 72.9%. We identified a non-coding control region of 327 bp according to its location and AT-richness. This is the smallest control region discovered in crustaceans so far. A mechanism of cytoplasmic tRNA import was addressed to compensate for the four missing tRNAs. The S. crosnieri mitogenome exhibits a novel arrangement of mitochondrial genes. We investigated the mitochondrial gene orders and found that at least six rearrangements from the ancestral pancrustacean (crustacean + hexapod) pattern have happened successively. The codon usage, nucleotide composition and bias show no substantial difference with other decapods. Phylogenetic analyses using the concatenated nucleotide and amino acid sequences of the 13 protein-coding genes prove consistent with the previous classification based upon their morphology. CONCLUSION: The present study will supply considerable data of use for both genomic and evolutionary research on hydrothermal vent ecosystems. The mitochondrial genetic characteristics of decapods are sustained in this case of S. crosnieri despite the absence of several tRNAs and a number of dramatic rearrangements. Our results may provide evidence for the immigrating hypothesis about how vent species originate. PMID- 18510776 TI - Divergence of canonical danger signals: the genome-level expression patterns of human mononuclear cells subjected to heat shock or lipopolysaccharide. AB - BACKGROUND: Peripheral blood mononuclear cells (PBMC) serve a sentinel role allowing the host to efficiently sense and adapt to the presence of danger signals. Herein we have directly compared the genome-level expression patterns (microarray) of a human PBMC model (THP-1 cells) subjected to one of two canonical danger signals, heat shock or lipopolysaccharide (LPS). RESULTS AND DISCUSSION: Based on sequential expression and statistical filters, and in comparison to control cells, we found that 3,988 genes were differentially regulated in THP-1 cells subjected to LPS stress, and 2,921 genes were differentially regulated in THP-1 cells subjected to heat shock stress. Venn analyses demonstrated that the majority of differentially regulated genes (> or = 70%) were uniquely expressed in response to one of the two danger signals. Functional analyses demonstrated that the two danger signals induced expression or repression of genes corresponding to unique pathways, molecular functions, biological processes, and gene networks. In contrast, there were 184 genes that were commonly upregulated by both stress signals, and 430 genes that were commonly downregulated by both stress signals. Interestingly, the 184 commonly upregulated genes corresponded to a gene network broadly related to inflammation, and more specifically to chemokine signaling. CONCLUSION: These data demonstrate that the mononuclear cell responses to the canonical stress signals, heat shock and LPS, are highly divergent. However, there is a heretofore unrecognized common pattern of gene network expression corresponding to chemokine-related biology. The data also serve as a reference database for investigators in the field of stress signaling. PMID- 18510777 TI - Improving the accuracy of Aboriginal and non-Aboriginal disease notification rates using data linkage. AB - BACKGROUND: Routinely collected infectious disease surveillance data provide a valuable means to monitor the health of populations. Notifiable disease surveillance systems in Australia have consistently reported high levels of completeness for the demographic data fields of age and sex, but low levels of completeness for Aboriginality data. Significant amounts of missing data associated with case notifications can introduce bias in the estimation of disease rates by population subgroups. The aim of this analysis was to evaluate the use of data linkage to improve the accuracy of estimated notification rates for sexually transmitted infections (STIs) and blood borne viruses (BBVs) in Aboriginal and non-Aboriginal groups in Western Australia. METHODS: Probabilistic methods were used to link disease notification data received in Western Australia in 2004 with core population health datasets from the established Western Australian Data Linkage System. A comparative descriptive analysis of STI and BBV notification rates according to Aboriginality was conducted based on the original and supplemented notification datasets. RESULTS: Using data linkage, the proportion of STI and BBV notifications with missing Aboriginality data was reduced by 74 per cent. Compared with excluding notifications with unknown Aboriginality data from the analysis, or apportioning notifications with unknown Aboriginality based on the proportion of cases with known Aboriginality, the rate ratios of chlamydia, syphilis and hepatitis C among Aboriginal relative to non Aboriginal people decreased when Aboriginality data from data linkage was included. CONCLUSION: Although there is still a high incidence of STIs and BBVs in Aboriginal people, incompleteness of Aboriginality data contributes to overestimation of the risk associated with Aboriginality for these diseases. Data linkage can be effectively used to improve the accuracy of estimated disease notification rates. PMID- 18510778 TI - Acute pancreatitis and subdural haematoma in a patient with severe falciparum malaria: case report and review of literature. AB - Plasmodium falciparum infection is known to be associated with a spectrum of systemic complications ranging from mild and self-limiting to life-threatening. This case report illustrates a patient who had a protracted course in hospital due to several rare complications of falciparum malaria. A 21-year old man presented with a five-day history of high-grade fever, jaundice and abdominal pain and a two-day history of altered conscious state. A diagnosis of severe falciparum malaria was made based on the clinical presentation and a positive blood smear with parasitaemia of 45%. Despite adequate anti-malarial therapy with artesunate, the patient had persistent and worsening abdominal pain. Investigations suggested a diagnosis of acute pancreatitis, a rare association with falciparum malaria. However, in spite of supportive therapy for acute pancreatitis and a 10-day course of intravenous artesunate and oral doxycycline at recommended doses, he continued to be febrile with peripheral blood smear showing persistence of ring forms. Antimalarial therapy was, therefore, changed to quinine on the suspicion of possible artesunate resistance. On the 17th day of stay in hospital, the patient developed generalized tonic-clonic seizures. Computerized tomography of the brain showed bilateral fronto-parietal subdural haematomas that were surgically drained. His fever persisted beyond 30-days despite broad-spectrum antibiotics, quinine therapy and negative malarial smears. A possibility of drug fever was considered and all drugs were ceased. He subsequently became afebrile and was discharged on the 38th hospital admission day. Recognition of complications and appropriate management at each stage facilitated successful outcome. This report has been presented to highlight the occurrence of several rare complications of falciparum malaria in the same patient. PMID- 18510779 TI - National quality and performance system for Divisions of General Practice: early reflections on a system under development. AB - BACKGROUND: Governments are increasingly introducing performance management systems to improve the quality and outcomes of health care. Two types of approaches have been described: assurance systems that use summative information for external accountability and internally driven systems that use formative information for continuous quality improvement. Australia recently introduced a National Quality and Performance System (NQPS) for Divisions of General Practice that has the dual purposes of increasing accountability and improving performance. In this article, we ask whether the framework can deliver on its objectives for achieving accountability and fostering performance improvement. We examine the system in terms of four factors identified in a recent systematic review of indicator systems known to improve their use. These are: involving stakeholders in development; having clear objectives; approach to data collection and analysis including using 'soft data' to aid interpretation; and feeding back information. RESULTS: We found that early consultative processes influenced system development. The system promotes the collection of performance information against defined program objectives. Data includes a mix of qualitative and quantitative indicators that are fitted to a conceptual framework that facilitates an approach to performance assessment that could underpin continuous quality improvement at the Division level. Feedback of information to support the development of quality improvement activities has not been fully developed. CONCLUSION: The system currently has elements that, with further development, could support a more continuous quality improvement or assurance based approach. Careful consideration needs to be given to the development of methods for analysis and review of performance indicators, performance assessment and engagement with consumers. The partnership arrangement that supported early development could be expected to serve as an important vehicle for further development. PMID- 18510780 TI - Wild leafy vegetables: a study of their subsistence dietetic support to the inhabitants of Nanda Devi Biosphere Reserve, India. AB - Consumption of greens is a major source of vitamins and micro-nutrients for people using only vegetarian diets rich in carbohydrates. In remote rural settlements where vegetable cultivation is not practiced and market supplies are not organized, local inhabitants depend on indigenous vegetables, both cultivated in kitchen gardens and wild, for enriching the diversity of food. Knowledge of such foods is part of traditional knowledge which is largely transmitted through participation of individuals of households. A total of 123 households in six villages of Nanda Devi Biosphere Reserve buffer zone was surveyed using a schedule to assess the knowledge, availability and consumption pattern of wild leafy vegetables. Quantity estimations were done using regular visits with informants from 30 sample households of the six study villages during the collections. Monetization was used to see the value of wild leafy vegetables harvested during a year. The diversity of wild leafy vegetables being use by the local inhabitants is 21 species belonging to 14 genera and 11 families. This is far less than that being reported to be used by the communities from Western Ghats in India and some parts of Africa. Irrespective of social or economic status all households in the study villages had the knowledge and used wild leafy vegetables. The number of households reported to consume these wild leafy vegetables is greater than the number of households reporting to harvest them for all species except for Diplazium esculentum and Phytolacca acinosa. The availability and use period varied for the species are listed by the users. The study indicated that the knowledge is eroding due to changing social values and non participation of younger generation in collection and processing of such wild leafy vegetables. PMID- 18510781 TI - Expression of nm23 in the spectrum of pre-invasive, invasive and metastatic breast lesions. AB - BACKGROUND: Nm23 protein is a metastasis suppressor protein, expressed in all tissues. Reduced Nm23 expression is related to a high incidence of lymph node and distant metastasis and poor prognosis in patients with cancers. The present study was done to analyze the expression of Nm23 using immunohistochemistry in non neoplastic and neoplastic breast lesions. METHODS: Sections from 93 samples were studied and classified into non-proliferative breast lesion (13), fibroadenoma (7), proliferative breast lesion (13), carcinoma in situ (20), invasive carcinoma (23) and metastatic deposits in lymph nodes (17). RESULTS: Nm23 expression in these groups showed a progressive down regulation with increasing neoplastic transformation. On comparing the various groups, nm23 expression was significantly different between the various subgroups with greatest expression in non-proliferative lesions and least in metastatic deposits (p < 0.050). CONCLUSION: It is concluded that the modulation of nm23 in a spectrum of breast lesions can be indicative of metastatic phenotype and help to predict the aggressiveness of disease. PMID- 18510782 TI - Carotid intima-media thickness and endothelial function: useful surrogate markers for establishing cardiovascular risk in patients with inflammatory rheumatic disease. PMID- 18510784 TI - Arthritis gene therapy's first death. AB - In July 2007 a subject died while enrolled in an arthritis gene therapy trial. The study was placed on clinical hold while the circumstances surrounding this tragedy were investigated. Early in December 2007 the Food and Drug Administration removed the clinical hold, allowing the study to resume with minor changes to the protocol. In the present article we collate the information we were able to obtain about this clinical trial and discuss it in the wider context of arthritis gene therapy. PMID- 18510785 TI - Process rather than pattern: finding pine needles in the coevolutionary haystack. AB - The geographic mosaic theory is fast becoming a unifying framework for coevolutionary studies. A recent experimental study of interactions between pines and mycorrhizal fungi in BMC Biology is the first to rigorously test geographical selection mosaics, one of the cornerstones of the theory. PMID- 18510786 TI - Recently published papers: a little less ventilation, a little more oxygen please? AB - Recent papers discussed include two large, multicentre, high-positive end expiratory pressure trials in acute lung injury and reflects upon the usefulness of such trial designs. Further papers considered include the emerging story of beta2-agonists for pulmonary oedema, highlights the newly described, iatrogenic demon, of ventilator-induced diaphragm injury, promotes the addition of B-type natriuretic peptide testing to the prediction of extubation success, and muses again over the oxygen debate. PMID- 18510787 TI - Defining and labelling 'healthy' and 'unhealthy' food. AB - OBJECTIVE: To consider the use of systematic methods for categorising foods according to their nutritional quality ('nutrient profiling') as a strategy for promoting public health through better dietary choices. METHODS: We describe and discuss several well-developed approaches for categorising foods using nutrient profiling, primarily in the area of food labelling and also with respect to advertising controls. The best approach should be able to summarise and synthesise key nutritional dimensions (such as sugar, fat and salt content, energy density and portion size) in a manner that is easily applied across a variety of products, is understandable to users and can be strictly defined for regulatory purposes. RESULTS: Schemes that provide relative comparisons within food categories may have limited use, especially for foods that are not easily categorised. Most nutrient-profiling schemes do not clearly identify less-healthy foods, but are used to attract consumers towards products with supposedly better profiles. The scheme used in the UK to underpin the colour-coded 'traffic light' signalling on food labels, and the one used by the UK broadcasting regulator Ofcom to limit advertising to children, together represent the most developed use of nutrient profiling in government policy-making, and may have wider utility. CONCLUSION: Nutrient profiling as a method for categorising foods according to nutritional quality is both feasible and practical and can support a number of public health-related initiatives. The development of nutrient profiling is a desirable step in support of strategies to tackle obesity and other non communicable diseases. A uniform approach to nutrient profiling will help consumers, manufacturers and retailers in Europe. PMID- 18510788 TI - The 'Sydney Principles' for reducing the commercial promotion of foods and beverages to children. AB - A set of seven principles (the 'Sydney Principles') was developed by an International Obesity Taskforce (IOTF) Working Group to guide action on changing food and beverage marketing practices that target children. The aim of the present communication is to present the Sydney Principles and report on feedback received from a global consultation (November 2006 to April 2007) on the Principles. The Principles state that actions to reduce marketing to children should: (i) support the rights of children; (ii) afford substantial protection to children; (iii) be statutory in nature; (iv) take a wide definition of commercial promotions; (v) guarantee commercial-free childhood settings; (vi) include cross border media; and (vii) be evaluated, monitored and enforced. The draft principles were widely disseminated and 220 responses were received from professional and scientific associations, consumer bodies, industry bodies, health professionals and others. There was virtually universal agreement on the need to have a set of principles to guide action in this contentious area of marketing to children. Apart from industry opposition to the third principle calling for a statutory approach and several comments about the implementation challenges, there was strong support for each of the Sydney Principles. Feedback on two specific issues of contention related to the age range to which restrictions should apply (most nominating age 16 or 18 years) and the types of products to be included (31% nominating all products, 24% all food and beverages, and 45% energy-dense, nutrient-poor foods and beverages). The Sydney Principles, which took a children's rights-based approach, should be used to benchmark action to reduce marketing to children. The age definition for a child and the types of products which should have marketing restrictions may better suit a risk-based approach at this stage. The Sydney Principles should guide the formation of an International Code on Food and Beverage Marketing to Children. PMID- 18510789 TI - [Factors associated with untreated hypertension among older adults: results of the Mexican Health and Aging Study, 2001]. AB - OBJECTIVE: To determine factors associated with failure to adhere to treatment for diagnosed hypertension among a representative sample of older Mexican adults living in the community. METHODS: A cross-sectional study of 2,029 individuals 65 years of age or older with diagnosed hypertension who participated in the Mexican Health and Aging Study, carried out during the summer of 2001. The survey collected information on several demographics (age, sex, schooling, whether living alone, and employment status, among others), any chronic illnesses, symptoms of depression, cognitive deterioration, body mass index, smoking, alcohol consumption, and difficulty performing basic and instrumental activities of daily living. Simple and multifactorial logistical regression analyses were used to evaluate the association among the study variables and self-reported untreated high blood pressure. RESULTS: Of the 2,029 participants, 437 (21.5%) reported not following any treatment whatsoever for controlling their hypertension; 1,584 (78.1%) affirmed they were complying with treatment; and 8 (0.4%) did not respond to this question. The multifactorial analysis adjusted for confounding variables (age, sex, symptoms of depression, and cognitive deterioration) showed that only a low number of years of schooling (adjusted odds ratio [OR] = 1.70; 95% confidence interval [95%CI]: 1.10-2.64; P = 0.02 for 1-6 years of schooling and adjusted OR = 3.32; 95%CI: 2.10-5.24; P < 0.01 for no schooling), alcohol consumption (adjusted OR = 1.52; 95%CI: 1.14-2.03; P = 0.01), and urinary incontinence (adjusted OR = 1.61; 95%CI: 1.15-2.26;P < 0.01) were independently associated with hypertension. CONCLUSIONS: Hypertension is a common and important issue among older adults in Mexico. To obtain better medication compliance, doctors prescribing or modifying hypertension treatment should taken into account whether or not the patient suffers from urinary incontinence, consumes alcohol, and/or has a low level of schooling. PMID- 18510790 TI - Universal vaccination of children against hepatitis A in Chile: a cost effectiveness study. AB - OBJECTIVE: To evaluate the healthcare and economic impact of routine hepatitis A vaccination of toddlers in Chile. METHODS: We used a dynamic model of hepatitis A infection to evaluate the impact of a two-dose vaccination program, administered at ages 12 and 18 months. The model incorporated the changing epidemiology of hepatitis A in Chile and the development of vaccine-induced herd immunity. Our analysis was conducted from the public payer perspective, and an estimation of the societal perspective was performed. Costs are expressed in 2005 U.S. dollars. RESULTS: Vaccination of toddlers rapidly reduced the healthcare burden of hepatitis A. In the base case (95% vaccination coverage, 100-year time horizon, 1% annual decrease in force of infection), the average number of infections fell by 76.6% annually, and associated deaths fell by 59.7%. Even at 50% coverage, the program reduced infection rates substantially. Routine vaccination of toddlers had economic as well as health benefits, saving $4,984 per life-year gained (base case scenario). The program became cost saving after 6 years, and its overall cost-effectiveness per life-year gained was largely unaffected by changes in disease-related costs, herd immunity, coverage rate, and annual decrease in force of infection. CONCLUSIONS: Routine vaccination of toddlers will reduce the rates of symptomatic hepatitis A and associated mortality. The two-dose schedule evaluated here will be less expensive than disease-related costs in the absence of vaccination from the sixth year of its implementation. These findings support the establishment of a routine vaccination program for toddlers in Chile. PMID- 18510791 TI - [Paracoccidioidomycosis-related mortality trend, state of Sao Paulo, Brazil: a study using multiple causes of death]. AB - OBJECTIVE: To investigate mortality in which paracoccidioidomycosis appears on any line or part of the death certificate. METHOD: Mortality data for 1985-2005 were obtained from the multiple cause-of-death database maintained by the Sao Paulo State Data Analysis System (SEADE). Standardized mortality coefficients were calculated for paracoccidioidomycosis as the underlying cause-of-death and as an associated cause-of-death, as well as for the total number of times paracoccidioidomycosis was mentioned on the death certificates. RESULTS: During this 21-year period, there were 1 950 deaths related to paracoccidioidomycosis; the disease was the underlying cause-of-death in 1 164 cases (59.69%) and an associated cause-of-death in 786 (40.31%). Between 1985 and 2005 records show a 59.8% decline in the mortality coefficient due to paracoccidioidomycosis as the underlying cause and a 53.0% decline in the mortality as associated cause. The largest number of deaths occurred among men, in the older age groups, and among rural workers, with an upward trend in winter months. The main causes associated with paracoccidioidomycosis as the underlying cause-of-death were pulmonary fibrosis, chronic lower respiratory tract diseases, and pneumonias. Malignant neoplasms and AIDS were the main underlying causes when paracoccidioidomycosis was an associated cause-of-death. The decision tables had to be adapted for the automated processing of causes of death in death certificates where paracoccidioidomycosis was mentioned. CONCLUSIONS: Using the multiple cause-of death method together with the traditional underlying cause-of-death approach provides a new angle on research aimed at broadening our understanding of the natural history of paracoccidioidomycosis. PMID- 18510792 TI - Severity of alcohol use and problem behaviors among school-based youths in Puerto Rico. AB - OBJECTIVES: The present study sought to: (a) categorize youths into groups based on their level of alcohol use and number of symptoms of alcohol abuse and dependence defined in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV), and (b) examine whether these categories were associated with other problem behaviors in which youths engage (marijuana use, sexual intercourse, and having been arrested or having trouble with the law). METHODS: The study is based on a cross-sectional survey administered to 972 school-based youths from one middle school and one high school in San Juan, Puerto Rico. Youths were categorized based on their alcohol use and alcohol problems. These categories were then examined for associations with lifetime marijuana use, lifetime sexual intercourse, and having been arrested or having had trouble with the law in the past year. The original eight categories of alcohol use were collapsed into six categories based on the results. RESULTS: For virtually every group characterized by higher severity of alcohol use and alcohol problems, researchers found an increasing prevalence of marijuana use in their lifetimes, increasing odds of sexual intercourse in their lifetimes, and having had trouble with the law in the past year. CONCLUSIONS: Knowing about variations in alcohol use and alcohol problems may be instrumental in measuring the degree to which youths may also be engaging in a range of other elevated risk behaviors and a progression to more serious forms of alcohol and drug use. PMID- 18510793 TI - [Risk factors for symptoms of allergic rhinitis in adolescents in Castellon, Spain]. AB - OBJECTIVE: To estimate the risk factor for symptoms of allergic rhinitis in adolescents in Castellon, Spain. METHODS: A cross-sectional population based study of Phase III of the International Study of Asthma and Allergies in Childhood (ISAAC) conducted in 2002 among adolescents from 13-14 years of age. The ISAAC questionnaire was used to define cases of allergic rhinitis. Logistic regression models were used to estimate odds ratio (OR) and their 95% confidence intervals (95%CI). RESULTS: Participation was 66.8% (3,995 adolescents of the 5,981 total). The prevalence of rhinoconjunctivitis symptoms in the last 12 months was 16.5% and the prevalence of nasal allergy at some point was 7.4%. Logistic regression showed that rhinoconjunctivitis was associated with being female (OR = 1.63; 95%CI: 1.33-2.00); a mother who smokes in the home (OR = 1.32; 95%CI: 1.08-1.63); a history of sinusitis (OR = 2.02; 95%CI: 1.51-2.70); and living on a street with heavy truck traffic (OR = 1.58; 95%CI: 1.02-2.44). Likewise, nasal allergy was associated with a family history of allergic rhinitis (OR = 2.62; 95%CI: 1.90-3.63); a history of sinusitis (OR = 2.65; 95% CI: 1.77 3.96); a history of bronchitis (OR = 1.68; 95%CI: 1.19-2.36); and social class, with a steady decline when comparing higher classes to lower classes. CONCLUSIONS: Various environmental risk factors were associated with the symptoms of nasal allergies, which points to the importance of implementing specific preventive measures. PMID- 18510794 TI - HIV risk behaviors of Latin American and Caribbean men who have sex with men in Miami, Florida, USA. AB - OBJECTIVE: The goal of this study is to describe the sexual practices, drug use behaviors, psychosocial factors, and predictors of unprotected anal intercourse (UAI) in a sample of Hispanic men who have sex with men (MSM) born in Latin American and Caribbean (LAC) countries who currently reside in Miami-Dade County, Florida. METHODS: Hispanic MSM (N = 566) recruited from community and Internet venues completed a computer-assisted self-interview assessing sociodemographic factors, drug use, sexual behaviors, and psychosocial factors. We focused on the 470 men who were born in LAC countries, including Puerto Rico. We first examined separately, by country of origin, the sexual practices, drug use behaviors, and psychosocial factors of the sample. We then collapsed the groups and examined the factors associated with UAI in the previous 6 months for the entire sample of Hispanic MSM from LAC countries. RESULTS: In the previous 6 months, 44% of the sample engaged in UAI, and 41% used club drugs. At the multivariate level, psychological distress, higher number of sexual partners, club drug use, HIV positive status at the time of immigration, and greater orientation to American culture were significantly associated with UAI in the previous 6 months. CONCLUSION: Many MSM born in LAC countries engage in HIV-related risk behaviors in the AIDS epicenter of Miami-Dade County, Florida. Culturally appropriate interventions should address these risk behaviors in this underserved population. PMID- 18510795 TI - [Underlying cause-of-death mortality statistics: considering the reliability of data]. AB - Mortality statistics are used in epidemiology and public health as an indicator of health status, to evaluate health programs, and in population studies to compare trends and spatial differences. One of the variables used in this type of analysis is the underlying cause-of-death. However, the quality of cause-of-death statistics based on the information recorded by physicians in death certificates has been criticized. The aim of this paper is to discuss the reliability of cause of-death data recorded by physicians in death certificates, based on studies carried out according to various methodologies, and to comment on the validity of using such underlying cause-of-death statistics. PMID- 18510796 TI - [How to implement Chile's new anti-tobacco law within the university]. AB - On the 16th of May 2006, the Diario Oficial de la Republica de Chile (Official News of the Republic of Chile), published Law No. 20.105, which modified Law No. 19.419 regarding advertising and consumption of tobacco. Since it entered into force, different institutions and public spaces have had to enact its regulations. The purpose of this article is to share how the new anti-tobacco law is being implemented, with a health promotion angle, by the Pontificia Universidad Catolica de Chile (UC). As such, we highlight key findings from a review of the literature on the subject and describe the implementation strategies developed and substantiated in theory as well as by how they address the concerns of the university community. The main accomplishments of the anti tobacco implementation were: reaching a compromise between the university directorate and the student leaders, who in turn motivated the student body to participate during the large-scale roll-out of the new law; the participation of all of the UC stakeholders; utilizing the varied methods of mass communication; and lastly, providing an opportunity for reflection and analysis for those directly affected: the smokers. The challenge is to evaluate the strategies implemented in the incorporation of the new Chilean anti-tobacco law. PMID- 18510797 TI - No association of defined variability in leptin, leptin receptor, adiponectin, proopiomelanocortin and ghrelin gene with food preferences in the Czech population. AB - BACKGROUND: Previously, it has been reported that mutations in the genes encoding for adipokines may be associated with impaired food intake and may serve as potential obesity biomarkers. The aim of this study was to investigate the possible associations of defined variability in leptin, leptin receptor, adiponectin, proopiomelanocortin and ghrelin genes with food preferences in the obese and non-obese Czech population and evaluate their potential as the obesity susceptibility genes. PATIENTS AND METHODS: Using PCR followed by restriction analysis, we studied 185 volunteers. Basic anthropometrical characteristics associated to obesity were measured and the food intake was monitored using a 7 day record method. In the group of obese individuals, a subset of 34 morbidly obese patients was studied for plasma leptin and soluble leptin receptor levels. RESULTS: None of the examined polymorphisms was associated to anthropometrical or demographic characteristics of the study subjects. The Gln223Arg polymorphism within the leptin receptor gene was significantly associated with lower plasma leptin levels (the RR genotype being more frequent in patients with lower plasma leptin levels; P = 0.001). No associations of the examined polymorphisms with food preferences was observed. CONCLUSIONS: Based on our results, the examined polymorphisms in the adipokine genes do not seem to be the major risk factor for obesity development in the Czech population nor significantly affect food preferences. PMID- 18510798 TI - Altered amino acid excretion in children with autism. AB - Autism is a complex and life-long behavioural disorder of unknown aetiology. Recent reports have indicated the involvement of digestive tract dysfunction and possible complications from inadequate nutrition. In this study, 34 autistic children (12 untreated and 22 receiving therapeutic treatments related to digestive function and nutritional uptake) and 29 control subjects (all 5-15 years of age) were investigated to determine whether there were any anomalies in the urinary excretion of amino acids, glucose, sucrose, arabinose and tartaric acid using GC/FID and GC/MS analysis techniques. Significantly lower relative urinary levels of essential amino acids were revealed for both the untreated (mean +/- SEM, 32.53 +/- 3.09%) and treated (31.98 +/- 2.87%) autistic children compared with the controls (37.87 +/- 1.50%). There were no significant differences in measured excretions of sugars or tartaric acid. It was concluded that the untreated autistic children had evidence of altered metabolic homeostasis. PMID- 18510799 TI - Screening for methylenetetrahydrofolate reductase C677T and A1298C polymorphisms in Indian patients with idiopathic mental retardation. AB - Although genetic, nutritional, and environmental factors have been found to aggravate mental retardation in approximately 1% of individuals, no cause is known till date. In this study, two genetic polymorphisms in methylenetetrahydrofolate reductase (MTHFR), C677T (rs#1801133) and A1298C (rs#1801131), have been investigated in idiopathic mental retardation (IMR) subjects. Significantly higher frequency of the C677 allele was observed in IMR (n = 155; chi(2) = 5.5; P = 0.019) and moderate IMR (n = 67; chi(2) = 6.16; P = 0.013) groups as compared to controls (n = 126); for A1298C, no significant difference was noticed. TDT analysis revealed preferential transmission of C677 allele to a small group of mild IMR probands (chi(2) = 5.545; P = 0.018). Higher frequency of CA haplotype was also noticed in IMR cases as compared to controls (chi(2) = 6.28; P = 0.012). We infer from the present investigation that these polymorphisms are not contributing to the aetiology of IMR in this population since both case-control and family-based analysis revealed no significant transmission of the mutated allele. PMID- 18510800 TI - Pseudotumor cerebri in pediatric age: role of obesity in the management of neurological impairments. AB - Pseudotumor cerebri occurs quite rarely in the pediatric population and its clinical features differ from adults in many ways. Intracranial hypertension with papilledema should obviously be treated promptly to avoid permanent visual damage, but various more or less invasive options have been proposed over the years, from bariatric surgery for obesity to optic nerve sheath fenestration. We report a prospective study on a group of 15 children, aged 3-16 years, with clinical and instrumental diagnosis of pseudotumor cerebri. All the patients were treated simply by external lumbar cerebrospinal fluid drainage with a mean volume of 10 ml/h for 3-5 days, with hypocaloric diet and with appropriate dosages of acetazolamide. All had immediate relief of headache, a considerable reduction in papilledema and marked improvement of both visual loss and cranial nerve palsies within 2 months. None of the patients relapsed during the follow-up period, ranging from 12-48 months. PMID- 18510801 TI - Consumption of green tea alters glial fibriliary acidic protein immunoreactivity in the spinal cord astrocytes of STZ-diabetic rats. AB - We examined the effect of green tea consumption on glial fibriliary acidic protein (GFAP) expression in spinal cord of streptozotocin (STZ) treated rats. Three groups (n = 10) were used in this study: (i) controls; (ii) STZ-induced diabetic rats given tap water; and (iii) an STZ-induced diabetic group given green tea. Immunohistochemistry showed a significant (P < 0.001) decrease in the number of GFAP immunoreactive astrocytes in spinal cord sections of diabetic rats compared to non-diabetic controls. Diabetic rats treated with green tea showed a significant (P < 0.01) increase in the number GFAP-immunoreactive astrocytes in all the spinal cord gray areas as compared to water-drinking diabetic rats. Immunoblotting confirmed that the diabetic spinal cord tissue expressed 71.0 +/- 7.0% less GFAP compared to non-diabetic controls and that the GFAP content in diabetic rats increased up to 86.34 +/- 18.74% compared to non-diabetic controls after 12 weeks of green tea consumption. In conclusion, consumption of green tea may represent an achievable adjunct therapy for improving changes seen in diabetic spinal cord. PMID- 18510802 TI - Suppressive effect of Yokukansan on excessive release of glutamate and aspartate in the hippocampus of zinc-deficient rats. AB - Yokukansan (TJ-54), a herbal medicine, has been used as a cure for insomnia and irritability in children. Yokukansan also improves behavioral and psychological symptoms such as agitation, aggression and irritability in patients with dementia including Alzheimer's disease, in which the glutamatergic neurotransmitter system is perturbed. However, the action of Yokukansan in synaptic neurotransmission is unknown. In the present study, the action of Yokukansan in the glutamatergic neurotransmitter system was examined in zinc-deficient rats, a neurological disease model, in which the glutamatergic neurotransmitter system is perturbed. Administration of Yokukansan significantly suppressed the increase in extracellular concentrations of glutamate and aspartate in the hippocampus after stimulation with 100 mM KCl, but not the increase in extracellular concentrations of glycine and taurine, suggesting that Yokukansan is involved in modulation of excitatory neurotransmitter systems. The present study demonstrates that Yokukansan is a possible medicine for prevention or cure of neurological diseases associated with excitotoxicity. PMID- 18510803 TI - Diet-induced obesity and spatial cognition in young male rats. AB - Recent work suggests that obesity may adversely affect cognitive behavior. To examine this suggestion, the effects of feeding a standard chow diet, and either supplemental sugar or fat on the development of obesity and performance on a test of spatial learning, the Morris Water Maze (MWM), were assessed in young male Long-Evans rats. Rats given access to a sucrose solution or dietary fat in addition to the chow diet consumed approximately 10% more calories per day, gained more weight, and had larger epididymal fat pads than rats fed the chow diet alone. Moreover, rats fed the supplemental sucrose took significantly more time to find a hidden platform in the MWM than rats fed the chow diet alone or chow and supplemental fat. Additionally, when tested 10 days after the initial training trials, rats given sucrose displayed deficits in long-term spatial memory. After 6 weeks on the diets, fasting blood glucose and serum triglyceride concentrations were significantly higher in sucrose-fed rats than in rats eating only the standard diet. These results indicate that diet-induced obesity resulting from excess sucrose intake, but not fat intake, in young animals impairs spatial learning and memory. It is hypothesized that these deficits arise from metabolic insults that leave the brain vulnerable to alterations in insulin sensitivity and glucose metabolism. PMID- 18510804 TI - Enterostatin up-regulates the expression of the beta-subunit of F(1)F(o)-ATPase in the plasma membrane of INS-1 cells. AB - Exposure to high-fat diet easily promotes overeating while at the same time disrupting insulin secretion and islet function. Enterostatin is a peptide which is secreted from the pancreas in response to high-fat feeding and has been shown to inhibit fat intake as well as insulin secretion in experimental animal models. Until recently, there was no known receptor for enterostatin. In 2002, Berger and co-workers found enterostatin to target the beta-subunit of the F(1)-ATPase in rat brain membranes as well as in a clonal beta-cell line (INS-1). In this study, we found the beta-subunit of F(1)-ATPase to be ectopically expressed in the plasma membrane of INS-1 cells using both immunohistochemistry and Western blotting. Incubation with enterostatin for 60 min resulted in a 3.5-fold increase of the protein expression of the beta-subunit of F(1)-ATPase in the plasma membrane. Furthermore, we found ATP to be able to displace the binding of enterostatin to purified bovine F(1)-ATPase. This reported targeting of enterostatin to the beta-subunit of F(1)-ATPase in insulin cells may provide a link between high-fat intake and islet function. PMID- 18510805 TI - Delayed CNS maturation in iron-deficient anaemic infants. AB - Direct evidence of CNS developmental alterations in iron-deficient anaemic (IDA) infants was obtained. Twenty 3-15-month-old IDA and 20 non-IDA infants (age and gender matched), healthy in every other respect, were studied. Complete blood and iron kinetics tests determined an IDA status. Psychomotor development was assessed through the test of Rogers and co-workers [Rogers SJ, Donovan CM, D'Eugenio D, Brown SL, Whiteside E, Moersch MS, Schafer DS. (eds) Developmental Programming for Infants and Young Children, Vol 2. University of Michigan Press, 1981] and under the 10-20 International System qEEG was performed (sleep/stage II). A Pearson's correlation test was applied between haematological, psychomotor and broad band EEG variables, and through ANOVA psychomotor and AP means were compared. IDA infants showed lower scores in cognition, fine motor and social/emotional areas, higher delta/theta and lower alpha power. Most correlations between haematological/psychological variables were positive. Delta/theta correlations were negative with self-care/gross and motor items while alpha/beta AP showed positive correlations with psychomotor and haematological variables. A clear association was found between EEG alterations and a low haematological/iron profile leading to a delayed psychomotor development. PMID- 18510806 TI - Cholecystokinin-58 is more potent in inhibiting food intake than cholecystokinin 8 in rats. AB - INTRODUCTION: Several cholecystokinin (CCK) forms have been detected in plasma, but most studies on food intake investigated the effects of CCK-8 only. Recently, it has been demonstrated that CCK-58 is the only endocrine-active form of CCK in rats. METHODS: CCK-58 was synthesized with a peptide synthesizer using FMOC chemistry and CCK-58 effects on food intake were compared to CCK-8 in rats. RESULTS: Both CCK-58 and CCK-8 inhibited food intake in a dose-dependent manner and were equally potent at 30 min. CCK-58 showed a prolonged inhibition of food intake compared to CCK8 at the higher dose tested (7 nmol/kg), inhibiting food intake also at 60 min, and cumulative food intake was inhibited for up to 210 min by CCK-58. CONCLUSIONS: CCK-58 has the same potency in inhibiting food intake as CCK-8 in rats, but inhibits food intake longer. This might be due to its tertiary structure resulting in a delayed plasma degradation or a prolonged binding at the CCK receptor. As CCK-58 is the major CCK form in the gut wall and possibly in the circulating blood in humans, the effects of CCK on food intake might have been underestimated in the past. PMID- 18510807 TI - Cognitive findings of an exploratory trial of docosahexaenoic acid and lutein supplementation in older women. AB - INTRODUCTION: Low dietary intake of docosahexaenoic acid (DHA) and/or foods rich in lutein may be associated with increased risk of cognitive decline in the elderly. SUBJECTS AND METHODS: The cognitive benefit of DHA and lutein in unimpaired elder women was explored in the context of a 4-month, double-blind, intervention trial of DHA and lutein supplementation for eye health. Forty-nine women (aged 60-80 years) were randomized to receive DHA (800 mg/day; n = 14), lutein (12 mg/day; n = 11), a combination of DHA and lutein (n = 14) or placebo (n = 10). Subjects underwent cognitive tests measuring verbal fluency, memory, processing speed and accuracy, and self-reports of mood at randomization and upon completion of the trial. RESULTS: Following supplementation, verbal fluency scores improved significantly in the DHA, lutein, and combined treatment groups (P < 0.03). Memory scores and rate of learning improved significantly in the combined treatment group (P < 0.03), who also displayed a trend toward more efficient learning (P = 0.07). Measures of mental processing speed, accuracy and mood were not affected by supplementation. CONCLUSIONS: These exploratory findings suggest that DHA and lutein supplementation may have cognitive benefit for older adults. PMID- 18510808 TI - Functional relationships between serum total cholesterol levels, executive control, and sustained attention. AB - This study investigated the potential relationship between serum total cholesterol (TC) and two specific aspects of cognition (executive control and sustained attention) in a non-elderly sample, after controlling for life stress and several sociodemographic and health variables. For each participant (n = 46), measurements of TC, physical health, and life stress were obtained, and executive control and sustained attention were assessed using the Tower of London and the Digit Vigilance Test. The outcomes of these cognitive assessments were correlated with TC, and a covariate-adjusted analysis was performed. After controlling for several co-variates, TC was found to be significantly negatively associated with components of executive control and sustained attention. Because these cognitive functions are crucial in the moment-to-moment regulation of behavior, elevated TC may have negative behavioral consequences in everyday life situations. PMID- 18510809 TI - The present status of human helminthic diseases in Iran. AB - Over the last few decades there have been several marked changes in the human helminthiases found in Iran. Fascioliasis is emerging as an important chronic disease of humans, especially in the northern province of Gilan (where outbreaks in 1989 and 1999 involved >7000 and >10,000 cases, respectively) and, more recently, in the western province of Kermanshah. In contrast, no cases of urinary schistosomiasis, a disease that once affected thousands of individuals in south western Khuzestan province, have been reported in Iran in recent years, and no cases of dracunculiasis have been seen in the country since the mid-1970s. Approximately 1% of all admissions to surgical wards are attributable to cystic echinococcosis, which is still considered endemic, but only a few cases of alveolar echinococcosis have been recorded. Over the last decade, there appears to have been a generally downward trend in the incidence of intestinal helminthiases in Iran. Recent estimates of the prevalences of ascariasis and strongyloidiasis, for example, lie between just 0.1% and 0.3%, and <1% of the population now appears to be infected with hookworm. In contrast, human infection with Hymenolepis and Enterobius remains relatively common. There have been a few case reports of toxocariasis and a few sero-epidemiological investigations of this disease but problems in accurate diagnosis have prevented good estimates of the general prevalence of this nematode infection. Just nine cases of pentastomiasis (all caused by Linguatula), 12 of dirofilariasis, one of gongylonemiasis, and three of moniliformiasis have been formally recorded in Iran. PMID- 18510810 TI - Predictors of serum ferritin and haemoglobin during pregnancy, in a malaria endemic area of western Kenya. AB - Between 2000 and 2004, a cross-sectional survey was conducted, as part of a prospective cohort study, among the women attending antenatal-care clinics in Bondo district, a malaria-endemic area of western Kenya. The aim was to assess the prevalence of iron deficiency and determine the predictors of haemoglobin and serum ferritin concentrations in the women who had a gestational age between 14 and 24 weeks. A standardized questionnaire was used to collect and store the relevant bio-data for the study. Haemoglobin and ferritin concentrations were evaluated, sickle-cell status was determined, and malarial parasitaemias were detected and evaluated, using blood samples collected at enrollment. Multiple regression analysis was then used to test for significant predictors of the haemoglobin and serum ferritin concentrations. Although 842 women were enrolled in the prospective cohort study, haemoglobin concentrations were evaluated for only 828 of them, serum ferritin levels for 621, and levels of parasitaemia for 812. The mean haemoglobin concentration recorded was 10.9 g/dl. Although 37.9% of the subjects had mild-moderate anaemia (7.0-10.5 g haemoglobin/dl), only 0.5% were severely anaemic (<7.0 g haemoglobin/dl). The geometric mean serum ferritin concentration recorded was 18.9 microg/litre, and 32.3% of the subjects evaluated had low serum concentrations of ferritin (<12 microg/litre). Among the parasitaemic primigravidae (but not the parasitaemic multigravidae), those found positive for sickle-cell trait had significantly lower haemoglobin concentrations than those found negative in a sickling test (P=0.01). Among the pregnant women of Bondo district, gravidity, malarial infection and sickle cell appear to be key predictors of haemoglobin concentration. PMID- 18510811 TI - Cryptosporidium species causing acute diarrhoea in children in Antananarivo, Madagascar. AB - A 13-month study of children presenting with acute diarrhoeal disease at hospitals and rehydration clinics in Antananarivo, Madagascar, was undertaken between May 2004 and May 2005. Cryptosporidiosis accounted for diarrhoea in 12 (5.6%) of the 215 children investigated. Cases of cryptosporidiosis were detected only in the rainy season, and the median age of cases was 13.5 months (range=1 day-27 months). As 11 of the cases of cryptosporidiosis were caused by Cryptosporidium hominis and only one by C. parvum, most of the cases were probably the result of anthroponotic transmission. GP60/45/15 gene polymorphisms indicated that the causative pathogens were of subtypes Ia, Id, Ie and IIc. PMID- 18510812 TI - Neurocysticercosis: detection of Taenia solium DNA in human cerebrospinal fluid using a semi-nested PCR based on HDP2. AB - Human neurocysticercosis (NC) is caused by Taenia solium larvae lodged in the central nervous system. This disease is usually diagnosed by radiology but the results are not always clear-cut and so immunological assays are often also used. A semi-nested PCR, based on the non-coding HDP2 sequence of T. saginata, has now been developed for detecting DNA from T. solium cysticerci and confirming NC. This PCR, which amplifies a 171-bp T. solium product, allowed the specific detection of just 174 attograms of T. solium DNA. The efficacy of the PCR was tested using cerebrospinal fluid (CSF) from neurological patients, including 46 confirmed Mexican cases of NC and 32 patients from non-endemic Spain. Eighteen of the confirmed cases [including 10 (71%) of the 14 with vesicular extraparenchymal cysticerci and four (17%) of the 24 with damaged cysticerci] and two (33%) of the six patients with 'uncertain' diagnosis (in whom a diagnosis of NC could not be established by radiological and immunological studies) were found PCR-positive. The 36 patients known to have neurological problems other than NC were found PCR negative. The HDP2 PCR offers a new tool in the diagnosis of NC and in exploring the pathogenesis of this serious disease. PMID- 18510813 TI - Population-based survey of taeniasis along the United States-Mexico border. AB - Taenia solium and T. saginata are zoonotic tapeworms of substantial medical and economic importance. Although human taeniasis is widely recognised as an endemic problem in Mexico, its presence in the United States is poorly understood. The first population-based study to estimate the prevalence of human infection with Taenia tapeworms along the Texas-Mexico border has recently been conducted. Households were interviewed in the Texan city of El Paso and in the neighbouring Ciudad Juarez, in Mexico. Faecal samples from household members were then checked for Taenia eggs by flotation and/or for Taenia copro-antigens in an ELISA. The overall prevalence of taeniasis in this border region was found to be 3% but, compared with the residents of Juarez, El Paso residents were 8.6-fold more likely to be tapeworm carriers. The interviews revealed some important differences between the two study sites, particularly the more frequent use of anthelminthic drugs on the Mexican side of the border. These findings have implications in terms of the planning of effective health-education campaigns to decrease the prevalence of taeniasis in the human populations along the Texas Mexico border. PMID- 18510814 TI - Missed treatment opportunities for schistosomiasis mansoni, in an active programme for the treatment of urinary schistosomiasis in Plateau and Nasarawa states, Nigeria. AB - Both Schistosoma haematobium and S. mansoni are endemic in Nigeria. Since 1999 the ministries of health of Plateau and Nasarawa states, assisted by The Carter Center, have provided mass drug administrations with praziquantel to villages where >20% of the school-aged children tested with urine dipsticks have been found to have haematuria (presumed to be caused by S. haematobium). The current extent of S. mansoni in Nigeria remains relatively unknown because the tests needed to detect human infection with this parasite are difficult to perform in many endemic areas. In a cross-sectional survey involving 924 children, the prevalence of S. mansoni was determined in 30 villages (in four local government areas) that had been excluded from mass praziquantel administrations because the prevalence of haematuria in their school-aged children had been found to be <20%. Seventeen (57%) of the surveyed villages had sufficient S. mansoni (i.e. prevalences of at least 10%) to warrant treatment. The results indicated that, if both S. haematobium and S. mansoni are taken into account, 81% of the villages in the four local government areas studied require treatment, compared with 50% if only S. haematobium is considered. At the moment, the costs of the village-by village diagnosis of S. haematobium and S. mansoni would be greater than those of the presumptive treatment of the school-aged children in all villages. Until improved and cheaper rapid diagnostic methods for S. mansoni become available, the cheapest approach to the overall problem of schistosomiasis in this part of Nigeria would therefore be wide-spread mass drug distributions, without screening for at-risk populations. PMID- 18510815 TI - Sero-epidemiology of toxocariasis in a rural settlement in Sao Paulo state, Brazil. AB - The seroprevalence of Toxocara canis and risk factors for infection with this parasite were explored in a rural settlement in Sao Paulo state, Brazil. Total IgA and IgE levels in 79 subjects were determined by turbidimetry and chemiluminescence, respectively. Total counts of leucocytes and erythrocytes and differential counts of leucocytes were made by flow cytometry. ELISA for the detection of anti-Toxocara IgG, IgA and IgE were standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis. Seventeen (21.5%) of the subjects were found positive for anti Toxocara IgG, with no significant differences in such seropositivity with age or gender. Thirty (38%) of the subjects showed eosinophilia and 70 (89%) had elevated levels of total IgE. Among the 17 subjects found seropositive for anti Toxocara IgG, the percentage of leucocytes represented by eosinophils (P=0.0069) and total levels of IgE (P=0.0452) were positively correlated with the levels of anti-TES IgE. Although anti-TES IgA was detected in 10 (59%) of the subjects, there was no significant correlation between the levels of total IgA and those of Toxocara-specific IgA. Only one of the 17 subjects found positive for anti Toxocara IgG had attended a secondary school and all but two belonged to households with monthly incomes of or=1x10(6) copies/ml and ALT levels between 1.5 to 10 times of upper limits of normal (ULN) were enrolled in the study. Ten mg/d of ADV was administered for 52 weeks. Line serum samples were collected for measuring HBV DNA and HBV markers. The efficacy of the treatment at week 52 was evaluated in patients with different ALT, HBeAg and HBV DNA levels at baseline and HBV DNA levels at week 12 after treatment. RESULTS: At week 52 of ADV treatment, the rates of HBV DNA<10(3) were 72.7%, 66.7% and 53.0% respectively in patients with ALT>5xULN, HBeAg350 s/co (30.2%, P<0.01) and HBV DNA>10(8) copies/ml (34.4%, P<0.05) at baseline. HBeAg seroconversion rates were 42.2% and 7.5% (P<0.01) in patients with HBeAg titer350 S/co at baseline. In patients with HBV DNA<10(3), 10(3)-10(5) and >10(5) copies/ml at week 12, the ratios of them with HBV DNA<10(3) less than 1000 copies/ml at week 52 were 82.6%, 57.1% and 17.5% and significant differences were found between these groups (P<0.05); HBeAg seroconversion rates were 52.2%, 25.7% and 5.0% (P<0.05); ALT normalization rates were 100%, 83% and 75%, significantly higher in patients with HBV DNA<10(3) copies/ml than those with HBV DNA>10(5) copies/ml (P<0.05) at week 12. HBV DNA and HBeAg seroconversion at week 52 correlated with HBV DNA levels at week 12 (r=0.6 and r=0.5 respectively, P<0.01). CONCLUSIONS: In HBeAg-positive CHB patients treated with adefovir dipivoxil, HBV DNA levels at week 12 can be used to predict the efficacy at week 52. HBV DNA<10(3) copies/ml at week 12 predict a better treatment result at week 52. PMID- 18510845 TI - [An investigation of clinical features and immunological functions of HCV specific cytotoxic T lymphocytes in HIV/HCV co-infected patients]. AB - OBJECTIVE: To investigate the clinical features, CD4+ T and CD8+ T cell counts, HIV RNA load, HCV RNA load, CD8+ T cell responses to HCV of HIV/HCV co-infected and HCV mono-infected patients and to assess the mutual influences of the two viruses in the infection. METHODS: Fifty-nine patients with HIV/HCV co-infection were enrolled in this study. Thirty-six patients with HCV mono-infection served as a comparison group. The liver function, peripheral blood CD4+ T and CD8+ T cell counts, HIV RNA load and HCV RNA load were compared between the groups. Peripheral blood mononuclear cells were analyzed by interferon-gamma ELISpot using a panel of HCV antigens. RESULTS: The frequency of HIV/HCV co-infection in those blood donors in Henan, China was 60.8%. ALT and AST in the HIV/HCV co infection patients were not different from those of the HCV group. Globulin in the HIV/HCV co-infection group was higher than that in the HCV group (P<0.01). CD4+ T cell counts in the HIV/HCV co-infection group were lower than those in the HCV group, but CD8+ T cell counts in the HIV/HCV co-infection group were higher than those in the HCV group (P<0.01). The HCV RNA loads were higher in the HIV/HCV co-infection group than in the HCV group(P<0.01). The magnitude of HCV specific CTL response to HCV-NS3 overlapping peptides in the HIV/HCV co-infection group (649.34+/-685.90) was higher than that in the HCV group (1233.70+/ 1085.16). Albumin was negatively correlated with HCV RNA (log10copies/ml) in the HIV/HCV co-infection group (r=-0.540). A positive correlation was found between platelet and peripheral blood CD4+ T cell counts (P<0.05). No linear correlation was found between HCV virus loads, HIV virus loads or peripheral blood CD4+ T cell counts. CONCLUSION: The frequency of HIV/HCV co-infection in the blood donors in Henan, China was 60.8%. HIV/HCV co-infection aggravated the progress of chronic hepatitis C. PMID- 18510846 TI - [Adefovir dipivoxil treatment of hepatic cirrhosis complicated with hepatitis B virus associated glomerulonephritis]. AB - OBJECTIVE: To investigate the efficacy and safety of adefovir dipivoxil (ADV) in treating hepatic cirrhosis complicated with hepatitis B virus associated glomerulonephritis. METHODS: Six hepatic cirrhosis (Child-Pugh A grade, liver function compensated) patients complicated with hepatitis B virus associated glomerulonephritis diagnosed by renal biopsy, real time PCR and urinary protein tests were treated with ADV for one year in addition to a routine treatment. The dosage of ADV was 100mg daily. RESULTS: After 3 and 6 months treatment the negative conversion rates of HBV-DNA were 33.3% and 83.3%; the negative conversion rates of HBeAg were 16.7% and 66.7%; the positive conversion rates of HBeAb were both 16.7%; the recovery rates of ALT were 83.3% and 100.0%; and the recovery rates of TBil were 66.7% and 83.3% respectively. Protein in the urine of two patients was decreased to 0.3 g/d and in three patients it was 50% of the original values. After 1 year treatment the disease subsided fully in 3 and partially in 2 patients. CONCLUSION: Treating hepatic cirrhosis complicated with hepatitis B virus associated glomerulonephritis using adefovir dipivoxil is effective and safe. PMID- 18510847 TI - [The effects of sympathetic neurotransmitters and adrenergic receptors on liver fibrosis in murine schistosomiasis]. AB - OBJECTIVE: To investigate the effects of sympathetic neurotransmitters and adrenergic receptors on liver fibrosis in murine schistosomiasis. METHODS: Mice were infestated with schistosoma by means of pasting cercariae on their abdomens. Thirty mice were randomly divided into a control group and a model group. Hematoxylin eosin and Van Gieson staining were used to view the histopathology of their livers. Immunofluorescence histochemistry and laser scanning confocal fluorescence microscopy were used to measure the a1A and beta2 adrenergic receptors in livers of the two groups of mice. High performance liquid chromatography-electrochemical detector (HPLC-ECD) was used to determine the concentration of norepinephrine (NE) and dopamine (DA) in the plasma of the mice. RESULTS: Immunofluorescence histochemistry showed that a1A and beta2 receptors were present in hepatocytes and hepatic sinusoids of the livers of the mice of the two groups, but there were many more in the livers of the schistosoma infected mice (t=-2.888; t=-6.648) (P<0.05). The results of HPLC-ECD showed that the levels of NE and DA in the model group were higher than those of the control group (t=-3.372; t=-4.428) (P<0.05). CONCLUSION: Sympathetic neurotransmitters and adrenergic receptors may participate in liver fibrogenesis in mice infected with schistosoma. PMID- 18510848 TI - [Tumor-targeting expression of a new tumor suppressor gene HCCS1 and its tumor selective inhibitory effects on hepatocellular carcinoma]. AB - OBJECTIVE: To construct a tumor-targeting recombinant adenovirus vector containing hepatocellular carcinoma suppressor gene HCCS1 to enhance the safety of tumor treatment. METHODS: CCK-8 assay was used to observe different inhibitory effects on normal and malignant liver cells with high expressions of HCCS1 protein. The relative transcriptional activity of PEG-3p was quantified by luciferase assay. Recombinant adenovirus Ad-PEG-3p-HCCS1 was packaged with AdEasy system and confirmed by PCR. The tumor-targeted expression of HCCS1 protein in cells infected with Ad-PEG-3p-HCCS1 was determined by Western blot. Crystal violet assay and MTT assay were applied to observe the selective anti-tumor effects of the newly constructed virus in vitro. RESULTS: A higher inhibitory rate of about 60% was found in BEL-7404 and SW-620 than that in L02 and NHLF 96 h after the high expression of HCCS1. Luciferase assay showed 3.9-, 4.7-, and 1.5 fold transcriptional activity in BEL-7404, BEL-7405 and QGY-7703 respectively, in comparison with that in L02. Ad-PEG-3p-HCCS1 was constructed successfully and was verified by PCR. Western blot indicated that high expression of HCCS1 could be induced in BEL-7404 and QGY-7703 but not in L02. Crystal violet assay and MTT assay showed that it remarkably reduced the toxicity to L02 but still had enough antitumoral effect on Ad-CMV-HCCS1. CONCLUSIONS: With high expression of HCCS1 the tumor cells we used are being inhibited more. PEG-3p has the tumor-selective driving function in malignant liver cells. Our recombinant adenovirus Ad-PEG-3p HCCS1 can tumor-targeting induce HCCS1 expression in tumor cells, which can improve the safety of gene therapy with HCCS1. PMID- 18510849 TI - [Effects of paclitaxel loaded ultrasound contrast agents on cell cycles and ultrastructural features of HepG2 cells]. AB - OBJECTIVES: To investigate the effects of paclitaxel loaded ultrasound contrast agents on cell cycles and ultrastructural features of HepG2 cells. METHODS: HepG2 cells were cultured, and divided into a blank control group, a paclitaxel group, an ultrasound contrast agents group, and a paclitaxel loaded ultrasound contrast agents group. Cell cycles of the four groups were detected by flow cytometry, and the ultrastructural changes of the cells were observed under a transmission electron microscope. RESULTS: Paclitaxel loaded ultrasound contrast agents blocked the HepG2 cells at their G2/M phases, and it also induced more apoptosis of the HepG2 cells. CONCLUSIONS: Paclitaxel loaded ultrasound contrast agents can block HepG2 cells at the G2/M phase and induce apoptosis of the cells. PMID- 18510850 TI - [Effect of siRNA targeted against survivin on the malignant behaviors of human hepatoma cells]. AB - OBJECTIVE: To study survivin expression in human hepatoma cells and the effects of survivin siRNA on the malignant phenotypes of human hepatocellular cell line HCCLM6. METHODS: Four hepatocellular carcinoma (HCC) cell lines were used. Semi quantitative RT-PCR and Western blot were used to measure and compare their survivin expressions. The siRNA expression vector pshRNA-survivin targeting the mRNA of survivin and vector pGPU6/GFP/Neo-NC (as a control) were constructed, and then transfected into HCCLM6 cells. FQ-PCR was used to quantify the mRNA levels of survivin. The malignant phenotypes of transfected HCCLM6 cells, including invasive activities and adhesive capabilities, were analyzed. RESULTS: Survivin expression gradually increased with the increase of the invasion and metastasis behaviors of the four HCC cell lines (P<0.05). The expression of survivin was highest in cell line HCCLM6. Survivin mRNA level was decreased by 93.500%+/ 3.117% after the pshRNA-survivin transfection. The cell adhesion rates significantly decreased in the cells transfected with pshRNA-survivin (cell adhesion rates were 11.403%+/-1.256% vs 32.545%+/-1.367%, t=20.732, P<0.01). The migrating number of HCCLM6 cells (13.5+/-0.9) transfected with pshRNA-survivin was also significantly decreased (t=14.5, P<0.01) as compared with the control group (32.6+/-1.4). CONCLUSION: The expression of survivin in HCC might have a close relationship to their invasion and metastasis properties. Sequence-specific shRNA can significantly reduce the survivin expression in the HCCLM6 cell line. Suppression of survivin expression in HCCLM6 cells transfected with pshRNA survivin can reduce their invasive and adhesive capabilities. PMID- 18510851 TI - [Clinical features of 107 autoimmune hepatitis patients and 30 of them with AIH primary biliary cirrhosis overlap syndrome]. AB - OBJECTIVE: In order to provide a reliable basis for the diagnosis and treatment of autoimmune hepatitis (AIH) and its overlap syndrome, we investigated the clinical, immunological characteristics of and the therapeutic methods for AIH and AIH-primary biliary cirrhosis (PBC) overlap syndrome. METHODS: One hundred seven patients (77 with AIH and 30 with AIH-PBC overlap syndrome) were enrolled in the study. Their clinical manifestations, serum liver function tests (LFTs) findings, serum immunoglobulins, liver histopathological changes and their responsiveness to the therapies were investigated. RESULTS: The age distribution of AIH patients showed a single peak during their fifties and their main clinical manifestations were malaise, abdominal distension, anorexia and jaundice. Serum gamma globulin and IgG were significantly higher than their normal levels. 74% of the patients were positive for anti-nuclear antibody (ANA), 32% of the patients were positive for anti-smooth muscle antibody (AMA), and over 50% of the patients suffered from concurrent extrahepatic autoimmune diseases. The main histological changes in the liver biopsies were interface hepatitis (65%), lobular hepatitis and rosette formation of liver cells. Bridging necrosis was observed in severe AIH cases. In the AIH-PBC overlap syndrome patients, the levels of serum ALT, AST, GGT, ALP and incidences of ANA and AMA/AMA-M2 were all significantly higher than those of the AIH group. After treating AIH patients with prednisolone and azathioprine (Aza), complete response was seen in 42 cases (70%), sustained response was seen in 26 cases (43%). Sixteen cases had relapses after the withdrawal of the treatment or prednisolone dosage was reduced lower than 10 mg/d. The cases having normal serum ALT, AST, gamma-globulin and IgG levels after treatment were still responding to the reduced prednisolone dosage of 5-10 mg/d without azathioprine added. After combination with ursodeoxycholic acid (UDCA) treatment, the liver function tests (AST, ALT, TBil) of AIH-PBC overlap syndrome patients also significantly improved compared to those before the treatment (P<0.01). CONCLUSION: AIH and AIH-PBC overlap syndrome are not rare in our clinics. Their diagnoses should be based on the clinical presentations, biochemical and immunological indices and liver histological changes. In AIH cases, once their AST, ALT, gamma-globulin and IgG levels return to normal, the prednisolone dosage can be maintained at 5-10 mg/d and Aza can even be withdrawn. Good improvement for patients with AIH-PBC overlap syndrome can be obtained with UDCA and immunosuppression treatment. PMID- 18510852 TI - [Clinical and pathological features of 16 patients with Gilbert syndrome and 2 cases with genetic analysis]. AB - OBJECTIVE: To summarize the clinical and pathological features of Gilbert syndrome. METHODS: The clinical features and liver histological findings of 16 cases of Gilbert syndrome were reviewed. RESULTS: Of the 16 cases (13 males and 3 females, with an age range from 14 to 40 years), all had recurrent jaundice, unconjugated hyperbilirubinemia and lipofuscin granules in the hepatocytes around the hepatic perivenular areas. The genetic analysis of the two patients showed that the site of genetic mutations were located at exon 1 (Gly71Arg). CONCLUSIONS: The diagnosis of Gilbert disease can be improved by combining the data of clinical features, the genetic analysis findings and the histological changes of the livers of the patients. PMID- 18510853 TI - [Association of hepatic lipase gene promoter polymorphism -514C/T with nonalcoholic fatty liver disease]. AB - OBJECTIVE: To study the prevalence of the hepatic lipase gene (LIPC) promoter polymorphism (at position -514) in patients with nonalcoholic fatty liver disease (NAFLD), and its relationship with the susceptibility to NAFLD. METHODS: Genotype of LIPC promoter was detected with PCR-RFLP in 106 patients with NAFLD. Body mass index, waist-to-hip ratio (WHR), blood pressure, CHOL, HDL, LDL, TG, FPG and FINS of the patients were measured. Index of insulin resistance was determined using the homeostasis model assessment (HOMA) method. One hundred six healthy subjects matched for age and sex served as controls. RESULTS: The frequency of CC genotype and C allele in the NAFLD group were significantly higher than those in the control group (31.1% vs 26.4%, 62.7% vs 54.2%, P<0.05). Compared with TT genotype, both CC genotype and CT genotypes had higher relative risk of NAFLD (OR: 3.73, 95% CI: 1.31, 10.63; OR: 3.60, 95% CI: 1.35, 9.60). At the same time, the non-carriers of T allele in -514 had higher WHR than the T carriers (0.877+/ 0.06 vs 0.848+/-0.06, t=2.072, P<0.05)). Logistic regression analysis showed that T substitution in LIPC-514 position (OR: 1.28, 95% CI 0.10-0.74) had a lower susceptibility to NAFLD. CONCLUSION: The LIPC-514C/T polymorphism is associated with WHR, and the T substitution of LIPC-514 may lower the susceptibility to NAFLD. PMID- 18510854 TI - [Role of caveolin-1 down-regulation by iRNA in human hepatocyte proliferation]. AB - OBJECTIVE: To investigate the role of caveolin-1 down-regulation in human hepatocyte proliferation in vitro. METHODS: The expression vector psiRNA-CAV1 was constructed and transfected into Chang liver cells (CHL). The caveolin-1 down regulated cell clones were selected by the antibiotic zeocin. The proliferation of the cell strain CAV7 was examined by MTT, in which untransfected CHL and HepG2 cells were set as controls. Expression of caveolin-1, Akt, Erk1/2, p-Akt and p Erk1/2 in the transfected and control cells was detected by Western blot. RESULTS: After caveolin-1 expression was down-regulated by RNAi, CHL increased faster at first (24 h and 72 h, P<0.05; 96 h, P<0.01), but slower later. P-Akt and p-Erk1/2 expressions were down-regulated, indicating that the growth and proliferation related Akt and Erk1/2 pathways were inhibited after caveolin-1 down-regulation. CONCLUSION: Caveolin-1 may play an important role in hepatocyte proliferation. PMID- 18510855 TI - [Effects of farnesoid X receptor ligand on the metabolism of bile acids in rats with estrogen-induced intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the effects and mechanism of farnesoid X receptor (FXR) and its ligands on the metabolism of bile acids in rats with estrogen-induced intrahepatic cholestasis of pregnancy (ICP). METHODS: An ICP rat model was established with estradiol benzoate (EB) injections. Then FXR ligand chenodeoxycholic acid (CDCA) was administrated (100 mg/kg daily) to ICP rats for 5 days. The serum TBA and expression of FXR and bile salt export pump (BSEP) in the rat livers were examined by immunohistochemistry and reverse transcription PCR. RESULTS: The levels of TBA in the CDCA group rats were significantly lower than the untreated rats [(17.2+/-4.1)micromol/L vs (29.3+/-6.4)micromol/L], and the expressions of mRNA and protein of FXR were significantly higher [(0.76+/ 0.09 vs 0.53+/-0.06, P<0.05 and 2.35+/-0.06 vs 1.83+/-0.05, P<0.017, respectively)], and the expressions of BSEP were also higher [(0.99+/-0.21 vs 0.76+/-0.07, P<0.017 and 1.88+/-0.03 vs 1.46+/-0.06, P<0.017, respectively)]. CONCLUSIONS: FXR plays an important role in modulating the metabolism of bile acids. CDCA can lower the levels of serum TBA by upregulating the expression of FXR and BSEP and then increasing the transport of the bile acids. These facts might present a new idea and target for the treatment of ICP. PMID- 18510856 TI - [Transforming growth factor beta 1 stimulates the expression of plasminogen activator inhibitor 1 in hepatic stellate cells]. PMID- 18510857 TI - [Clinical analysis of liver transplantation in elderly patients]. PMID- 18510858 TI - [The effect of exogenous PEG10 on mitochondrial membrane potential of L02 human liver cells and its possible mechanisms]. PMID- 18510859 TI - [The radiosensitization effect of cyclooxygenase-2 selective inhibitor NS-398 on HepG2 cells and its mechanism]. PMID- 18510860 TI - [Preventive and treatment effects of granulocyte colony stimulating factor on CCl4 induced chronic liver injury in mice]. PMID- 18510861 TI - [Relationship of the transforming growth factor beta 3/beta 1 mRNA ratio and collagen I synthesis in rat hepatic stellate cells]. PMID- 18510862 TI - [HBx protein and its down-streaming molecules in hepatocellular carcinomas]. PMID- 18510863 TI - Blood-neural barrier: its diversity and coordinated cell-to-cell communication. AB - The cerebral microvessels possess barrier characteristics which are tightly sealed excluding many toxic substances and protecting neural tissues. The specialized blood-neural barriers as well as the cerebral microvascular barrier are recognized in the retina, inner ear, spinal cord, and cerebrospinal fluid. Microvascular endothelial cells in the brain closely interact with other components such as astrocytes, pericytes, perivascular microglia and neurons to form functional 'neurovascular unit'. Communication between endothelial cells and other surrounding cells enhances the barrier functions, consequently resulting in maintenance and elaboration of proper brain homeostasis. Furthermore, the disruption of the neurovascular unit is closely involved in cerebrovascular disorders. In this review, we focus on the location and function of these various blood-neural barriers, and the importance of the cell-to-cell communication for development and maintenance of the barrier integrity at the neurovascular unit. We also demonstrate the close relation between the alteration of the blood-neural barriers and cerebrovascular disorders. PMID- 18510864 TI - Application of hybrid LRR technique to protein crystallization. AB - LRR family proteins play important roles in a variety of physiological processes. To facilitate their production and crystallization, we have invented a novel method termed "Hybrid LRR Technique". Using this technique, the first crystal structures of three TLR family proteins could be determined. In this review, design principles and application of the technique to protein crystallization will be summarized. For crystallization of TLRs, hagfish VLR receptors were chosen as the fusion partners and the TLR and the VLR fragments were fused at the conserved LxxLxLxxN motif to minimize local structural incompatibility. TLR-VLR hybridization did not disturb structures and functions of the target TLR proteins. The Hybrid LRR Technique is a general technique that can be applied to structural studies of other LRR proteins. It may also have broader application in biochemical and medical application of LRR proteins by modifying them without compromising their structural integrity. PMID- 18510865 TI - Multiple shRNA expressing vector enhances efficiency of gene silencing. AB - RNA interference (RNAi) is the process of sequence-specific gene silencing. However, RNAi efficiency still needs to be improved for effective inhibition of target genes. We have developed an effective strategy to express multiple shRNAs (small hairpin RNA) simultaneously using multiple RNA Polymerase III (Pol III) promoters in a single vector. Our data demonstrate that multiple shRNAs expressed from Pol III promoters have a synergistic effect in repressing the target gene. Silencing of endogenous cyclophilin A (CypA) or key HIV viral genes by multiple shRNAs results in significant inhibition of the target gene. PMID- 18510866 TI - Overexpression of a delayed early gene hlg1 of temperate mycobacteriophage L1 is lethal to both M. smegmatis and E. coli. AB - Two genes of temperate mycobacteriophage L5, namely, gp63 and gp64, were hypothesized to be toxic to M. smegmatis. An identical L5 gp64 ortholog (designated hlg1) was cloned from homoimmune mycobacteriophage L1 and characterized at length here. As expected, hlg1 affected the growth of M. smegmatis when overexpressed from a resident plasmid. HLG1 (the protein encoded by hlg1) in fact caused growth retardation of M. smegmatis and the region encompassing its 57-114 C-terminal amino acid residues was found indispensable for its growth-retardation activity. Both nucleic acid and protein biosynthesis were severely impaired in M. smegmatis expressing HLG1. Interestingly, HLG1 also affected E. coli almost similarly. This putative delayed early lipoprotein did not participate in the lytic growth of L1. PMID- 18510867 TI - Apoptosis-inducing effect and structural basis of Polygonatum cyrtonema lectin and chemical modification properties on its mannose-binding sites. AB - Polygonatum cyrtonema Lectin (PCL), which is classified as a monocot mannose binding lectin, has received great regards for its uniquely biological activities and potentially medical applications in cancer cells. This paper was initially aimed to study apoptosis of PCL on Hela cells. Thus, 3-(4, 5-dimethylthiazol-2 yl)-2, 5-diphenyltetrazolium bromide (MTT) method was carried out. Through observation of cell morphologic changes and Lactate dehydrogenase (LDH) activity based cytotoxicity assays, PCL induced HeLa cell apoptosis in a dose-dependent manner. To further gain structural basis, multiple alignments, homology modeling and docking experiments were performed to analyze the correlation between its biological activities and mannose-binding sites. Eventually, considering docking data, chemical modification properties on the three mannose-binding sites were analyzed by a series of biological experiments (e.g., hemagglutinating and mitogenic activity assays, fluorescence and Circular Dichrosim (CD) spectroscopy) to profoundly identify the role of some key amino acids in the structure-function relationship of PCL. PMID- 18510868 TI - RNA silencing-mediated resistance is related to biotic / abiotic stresses and cellular RdRp expression in transgenic tobacco plants. AB - The discovery of RNA silencing inhibition by virus encoded suppressors or low temperature leads to concerns about the stability of transgenic resistance. RNA dependent RNA polymerase (RdRp) has been previously characterized to be essential for transgene-mediated RNA silencing. Here we showed that low temperature led to the inhibition of RNA silencing, the loss of viral resistance and the reduced expression of host RdRp homolog (NtRdRP1) in transgenic T4 progeny with untranslatable potato virus Y coat protein (PVY-CP) gene. Moreover, RNA silencing and the associated resistance were differently inhibited by potato virus X (PVX) and tobacco mosaic virus (TMV) infections. The increased expression of NtRdRP1 in both PVX and TMV infected plants indicated its general role in response to viral pathogens. Collectively, we propose that biotic and abiotic stress factors affect RNA silencing-mediated resistance in transgenic tobacco plants and that their effects target different steps of RNA silencing. PMID- 18510869 TI - Heat stress protection in Aspen sp1 transgenic Arabidopsis thaliana. AB - It is known that the stable protein 1 (SP1) detected in aspen plants remains soluble upon boiling and that sp1 expression in transgenic aspen is resistant to salt stress. Presently, we analyzed the effect of expression of SP1 in Arabidopsis thaliana plants and their response to high temperature stress. After 45 degrees C for 16 h, relative to wild type plants, sp1 transgenic plants exhibited stronger growth and were better in several physiological properties including chlorophyll, chlorophyll fluorescence, water content, proline content, and malondialdehyde content. These preliminarily results suggest that the over expression of SP1 may notably enhance heat-tolerant level of transgenic A. thaliana plants. PMID- 18510870 TI - A heat shock cognate 70 gene in the endoparasitoid, Pteromalus puparum, and its expression in relation to thermal stress. AB - The Pphsc70 (heat shock cognate 70) gene was isolated from the endoparasitoid Pteromalus puparum and then characterized. The full-length cDNA was 2204 base pair (bp) and contained a single 1968 bp ORF that encoded a polypeptide of 656 amino acids with a predicted molecular mass of 71.28 kDa. Phylogenetic analysis based on Hsc70 amino acid sequences from fifteen insect species agreed with the present phylogeny. In addition, genomic DNA confirmed the presence of three introns located at the coding region as well as the 5'UTR. A significant elevation of Pphsc70 expression was observed following heat treatment, however, continued exposure to heat shock or recovery caused the expression of induced mRNA to gradually decline to levels that were significantly lower than those of control pupae (P < 0.05). In addition, a significant increase was observed in the emergence rate of pupae that were preheated at 40 degrees C and then exposed to 50 degrees C for 1 h when compared with the pupae that were not preheated, but instead directly exposed to 50 degrees C. Taken together, these results revealed that exposure to gradually increasing temperatures can enhance an insects thermo tolerance. PMID- 18510871 TI - Intrinsic bent DNA colocalizes with the sequence involved in the Nd-sD mutation in the Bombyx mori fibroin light chain gene. AB - Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd s(D) mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-s(D) mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene. PMID- 18510872 TI - Bombyx mori protein disulfide isomerase enhances the production of nuecin, an antibacterial protein. AB - The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpressions of foreign proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins. PMID- 18510873 TI - Soluble expression and purification of synthetic human bone morphogenetic protein 2 in Escherichia coli. AB - A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2. PMID- 18510874 TI - Human brain pyridoxal-5'-phosphate phosphatase (PLPP):protein transduction of PEP 1-PLPP into PC12 cells. AB - Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1 PLPP to media pretreated with the vitamin B(6) precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin B(6). PMID- 18510875 TI - A review and critical appraisal of measures of therapist-patient interactions in mental health settings. AB - OBJECTIVES: To assemble and to appraise critically the current literature on tests and measures of therapist-patient interactions in order to make recommendations for practice, training and research, and to establish benchmarks for standardisation, acceptability and routine use of such measures. DATA SOURCES: Major electronic databases (including PsycINFO) were searched from inception to 2002. REVIEW METHODS: A comprehensive conceptual map of the subject area of therapist-patient interactions was developed through data extraction from, and analysis of, studies selected from the literature searches. The results of these searches were assessed and appraised to produce a set of possible therapist-patient measures. These measures were then evaluated. RESULTS: The contextual map included the various concepts and domains that had been used in the context of the literature on therapist-patient interactions, and was used to guide the successive stages of the review. Three developmental processes were identified as necessary for the provision of an effective therapeutic relationship: 'establishing a relationship', 'developing a relationship' and 'maintaining a relationship'. Eighty-three therapist-patient measures having basic information on reliability and validity were identified for critical appraisal. The areas of the conceptual map that received most coverage (i.e. over 50% measures associated with them) were framework, therapist and patient engagement, roles, therapeutic techniques and threats to the relationship. These areas relate to the three key developmental processes outlined above. Of the 83 measures matching the content domain, 43 met the minimum standard. A total of 30 measures displayed adequate responsiveness or precision. None of the 43 measures that met the minimum standard was fully addressed in terms of acceptability and feasibility evidence. The majority of these measures had three or fewer components described. Therefore, out of a total of 83 measures matching the content domain, no measure could be said to have met an industry standard. CONCLUSIONS: The findings indicate that the therapist-patient interaction can be measured using a wide range of instruments of varying value. However, due care should be taken in ensuring that the measure is suitable for the context in which it is to be used. Following on from this work, it is suggested that specific research networks for the development of therapist-patient measures should be established, that research activity should prioritise investment in increasing the evidence base of existing measures rather than attempting to develop new ones, and that research activity should focus on improving these existing measures in terms of acceptability and feasibility issues. PMID- 18510876 TI - Clostridium difficile infection: a critical overview. AB - Clostridium difficile is a gram-positive, spore-forming, toxin-producing anaerobic bacillus identified as the causal agent of a variety of manifestations typically isolated to the colon, but in its severe form, it can lead to sepsis and death. C. difficile infection due to a toxin gene variant strain (BI/NAP1) has been identified at the center of outbreaks and has resulted in increased mortality. Many questions remain as to how this strain appeared so quickly and has harmed or killed so many patients. We present a review of C. difficile infection, discussing its clinical presentation, diagnosis, management, and prevention. PMID- 18510877 TI - Methicillin-resistant Staphylococcus aureus infections of the eyes. PMID- 18510878 TI - Methicillin-resistant Staphylococcus aureus infections of the sinuses. PMID- 18510879 TI - The effect of vaccination on Streptococcus pneumoniae resistance. AB - Following the introduction of the 7-valent pneumococcal conjugate vaccine (PCV-7) in the routine immunization schedule of US children, the morbidity and mortality associated with Streptococcus pneumoniae infections have changed considerably. Post-licensure data from the United States have confirmed that PCV-7 has significantly decreased the incidence of vaccine-type susceptible and antibiotic resistant invasive pneumococcal diseases, community-acquired respiratory infections, and nasopharyngeal colonization in vaccinated individuals and their contacts. An unintended consequence of immunization with PCV-7 is the simultaneous increase in the carriage rates of non-vaccine-serotype pneumococci (some of which are highly antibiotic-resistant) among vaccinated children (ie, replacement phenomenon). Neither the implications of increased non-vaccine-type colonization nor the additional role of inappropriate antibiotic use in this process is clearly understood, but the greatest concern is that replacement colonization may result in replacement disease. Future vaccination with PCV-7 in European countries could provide additional information. PMID- 18510880 TI - The prospect of vaccination against group A beta-hemolytic streptococci. AB - Group A streptococcus is a widespread human pathogen that causes a broad spectrum of human disease. The persistent high burden and severity of illness in developing and industrialized countries speaks to the need for a safe and effective vaccine. Modern approaches to vaccine construction include M protein type-specific vaccines, vaccines utilizing conserved M antigens, and vaccines based on other conserved surface-expressed or secreted antigens. Vaccine candidates in various stages of development offer promise for prevention of Group A streptococcal infections and their sequelae. PMID- 18510881 TI - Diagnosis and management of supraglottitis (epiglottitis). AB - Acute supraglottitis is a serious, potentially fatal infection in both adults and children. Haemophilus influenzae type b (Hib) is the most common cause in children, even in this era of the Hib vaccine. Most cases of acute supraglottitis in adults are thought to be caused by other bacterial organisms, viral or combined viral-bacterial infections, and noninfectious etiologies. Early visualization of the larynx is essential, as is establishment of a secure airway in selected cases. Most cases will respond to intravenous antibiotic therapy and will not require an artificial airway. Admission to an area of high-intensity nursing is essential. Daily laryngoscopy is necessary to monitor response to treatment. Although the incidence of acute supraglottitis in children has decreased, isolated cases will be encountered and demand high vigilance. PMID- 18510882 TI - Maxillary sinusitis of odontogenic origin. AB - Odontogenic etiology accounts for 10% to 12% of cases of maxillary sinusitis. Although uncommon, direct spread of dental infections into the maxillary sinus is possible due to the close relationship of the maxillary posterior teeth to the maxillary sinus. If a periapical dental infection or dental/oral surgery procedure violates the schneiderian membrane integrity, infection will likely spread into the sinus, leading to sinusitis. An odontogenic source should be considered in individuals with symptoms of maxillary sinusitis and a history of dental or jaw pain; dental infection; oral, periodontal, or endodontic surgery; and in those people resistant to conventional sinusitis therapy. An odontogenic infection is a polymicrobial aerobic-anaerobic infection, with anaerobes outnumbering the aerobes. Diagnosis requires a thorough dental and clinical evaluation including radiographs. Management of sinus disease of odontogenic origin often requires medical treatment with appropriate antibiotics, surgical drainage when indicated, and treatment to remove the offending dental etiology. PMID- 18510883 TI - Do statins and/or ACE inhibitors impact risk for or outcome of pneumonia in certain populations? PMID- 18510884 TI - The value of prognostic indices for pneumonia. AB - One of the most important decisions in the management of community-acquired pneumonia is deciding the care site, which affects morbidity, mortality, and costs. Clinical judgment alone is difficult and imprecise. The Pneumonia Severity Index score and the CURB-65 (confusion, urea nitrogen, respiratory rate, blood pressure, 65 years of age and older) score are validated prognostic indices to predict mortality, and they can identify low-risk patients who may be eligible for outpatient management. However, limitations of the scoring systems preclude their isolated use, and they can only be recommended as an aid to guide hospital admission decisions. The Pneumonia Severity Index score is slightly better at identifying the lowest risk patients, whereas CURB-65 is much simpler to use. As an adjunct to clinical judgment, we consider CURB-65 to be the most useful prognostic index for identifying low-risk patients. PMID- 18510885 TI - Pneumonia in elderly patients with chronic obstructive pulmonary disease. AB - This article reviews the association between pneumonia and chronic obstructive pulmonary disease (COPD) and the possible role of inhaled corticosteroids in increasing the risk of pneumonia in patients with COPD. An increased risk of pneumonia with inhaled corticosteroids was first reported from the Toward Revolution in COPD Health (TORCH) study, a large randomized clinical trial comparing fluticasone, salmeterol, or a combination of the two medications with placebo. We carried out a large observational study using a health care administrative database of information on hospitalizations and medication use among patients older than 65 years of age in Quebec. We found an excess of pneumonia requiring hospitalization and an excess of pneumonia hospitalizations leading to death in relation to current use of inhaled corticosteroids, especially at high doses. Here, we explore the potential mechanisms of this association and try to weigh the benefits and risks of therapy with inhaled corticosteroids in patients with COPD. PMID- 18510886 TI - The changing epidemiology of pneumococcal pulmonary disease in the era of the heptavalent vaccine. AB - Since the pediatric pneumococcal 7-valent conjugate vaccine was licensed in the United States, dramatic changes have taken place in the occurrence of pneumococcal disease among children and adults. Incidence of invasive pneumococcal disease including bacteremic pneumonia has declined, but changes in noninvasive pulmonary disease have been more difficult to assess. This paper reviews evidence of the epidemiologic features of invasive pneumococcal disease and community-acquired pneumonia among children and adults since 2000. Despite some increases in non-vaccine-type disease, its use has resulted in major reductions in incidence of invasive pneumococcal disease in all age groups, particularly that of antibiotic-resistant disease and of pediatric pneumococcal pneumonia. Several studies show that changes in circulating pneumococcal strains may be producing differential changes in disease incidence and disease presentation in several populations, including HIV-infected individuals and Alaska Natives. PMID- 18510887 TI - Complicated parapneumonic effusion and empyema: pleural decortication and video assisted thoracic surgery. AB - Complicated parapneumonic effusion or empyema, in which the fluid becomes more exudative, develops in 10% to 20% of patients with parapneumonic effusions. Early diagnosis and effective treatment strategies are required. New treatment strategies such as fibrinolytic therapy and decortication or debridement through video-assisted thoracoscopic surgery, have changed the choice of treatment for different stages of empyema or complicated pleural effusion. This review discusses the definition, clinical presentations, diagnosis, and treatment of these conditions, focusing on the value and comparison of video-assisted thoracoscopic surgery decortication and fibrinolytic therapies. PMID- 18510888 TI - Update on male circumcision: prevention success and challenges ahead. AB - This article reviews the findings of three trials of male circumcision for HIV prevention, with emphasis on the public health impact, cultural and safety concerns, implications for women, and the challenges of roll out. Three randomized trials in Africa demonstrated that adult male circumcision reduces HIV acquisition by 50% to 60%. As circumcision provides only partial protection, higher risk behaviors could nullify circumcision's effect. Additionally, circumcision among HIV-infected men does not directly reduce male-to-female HIV transmission among discordant couples, according to the results of a recent Ugandan study. The roll-out or full-scale implementation requires committed expansion into existing HIV prevention programs. Efforts should include attention to safety, implications for women, and risk compensation. Rapid, careful establishment of circumcision services is essential to optimize HIV prevention in countries with the highest prevalence. PMID- 18510889 TI - Regulatory T cells in HIV infection: who's suppressing what? AB - The role of regulatory T cells (Treg) in HIV pathogenesis is not fully elucidated. Persistent antigens such as HIV are believed to promote the expansion and activation of antigen-specific Treg, and several reports have described beneficial and detrimental roles for Treg in HIV pathogenesis. These apparently contradictory observations may arise from imprecision in enumerating Treg and the lack of definition of Treg subsets. New markers allowing more precise identification and purification of Treg for functional studies have been described recently, and these may open avenues for efficient isolation of pure, homogenous populations of human Treg. PMID- 18510890 TI - Pathogenesis and management of lipoatrophy. AB - Despite impressive decreases in mortality and morbidity, significant adverse events have surfaced as a result of combination antiretroviral therapy (ART). They include lipoatrophy, or subcutaneous fat wasting of the face, arms, buttocks, or legs, which can be associated with central fat accumulation. Although the underlying mechanism of ART-related body fat abnormalities has not been definitively established, mitochondrial toxicity is increasingly implicated in the lipoatrophy component of these fat abnormalities. Several studies evaluating switches off of nucleoside analogues have showed modest but statistically significant increases in limb fat. Because ART switches result in slow and small improvements and are not an option in many patients, other therapeutic interventions are needed. Although peroxisome proliferator-activated receptor chi agonist thiazolidinediones would be expected to have positive effects on lipoatrophy, initial clinical studies are conflicting. Other interventions of uridine, pravastatin, and facial fillers have been evaluated in small studies. PMID- 18510891 TI - Effects of antiretroviral therapy on cognitive impairment. AB - Since the introduction of combination antiretroviral therapy, the manifestations of HIV-associated central nervous system (CNS) involvement have generally become less severe and more manageable. Patients initiating antiretroviral therapy have demonstrated improvement in cognitive functioning. Reduction of viral burden has been correlated with improved CNS functioning. Recent studies have focused on the importance of CNS-penetrating antiretroviral regimens to target active replication in this protected compartment. There were initial conflicting reports of whether CNS-penetrating antiretrovirals improved cognitive functioning, with more evidence supporting the importance of CNS penetration in reducing viral burden and improving cognitive outcomes. Cognitive loss remains a feature of HIV infection, and some patients still suffer from incident or progressing cognitive and motor dysfunction even on combination therapy. Because the virus enters the CNS within days of initial infection, latent infection may be chipping away at the brain during the long asymptomatic period in HIV. When to start antiretrovirals for the maximum cognitive benefit remains to be determined. PMID- 18510892 TI - Syphilis: continuing public health and diagnostic challenges. AB - As the "great imitator" of disease, syphilis, caused by the bacterium Treponema pallidum, continues to be a conceptually elusive condition that is surrounded by diagnostic ambiguity and clinical misunderstanding. Concurrent HIV infection adds further difficulty by introducing the oldest and most confusing medical conundrum into the socially and biologically complex situation of treating patients with a virus we are still only learning to manage. Syphilis continues to be a challenge to the health of men who have sex with men and people of color as infection rates continue to increase within these substantial subgroups living with HIV. As the resurgence of syphilis continues, a clear understanding of diagnostic testing, disease-staging paradigms, and treatment strategies are necessary for optimal management of the HIV co-infected patient. PMID- 18510893 TI - Human papillomavirus and anal neoplasia. AB - Anal cancer is a rare disease in the general population, but the incidence of anal cancer is higher in certain at-risk groups, such as men who have sex with men (MSM), and immunosuppressed individuals, including those with HIV infection. Among HIV-positive MSM, the incidence of anal cancer may be as high as 10 times greater than current rates of cervical cancer in the general population of women. Anal cancer is associated with human papillomavirus (HPV) infection and may be preceded by high-grade anal intraepithelial neoplasia (HGAIN). HGAIN and anal HPV infection are both highly prevalent in groups at risk for anal cancer. Current issues include determining the effect of antiretroviral therapy on the natural history of HGAIN and the incidence of anal cancer, optimizing diagnostic and therapeutic approaches to HGAIN, and determining the potential for prophylactic HPV vaccines to prevent anal HPV infection and anal cancer in at-risk groups. PMID- 18510895 TI - Scaling up ART treatment capacity: lessons learned from South Africa, Zimbabwe, and Botswana. AB - Over the past 3 years, significant strides have been made in the effort to provide antiretroviral therapy (ART) to the millions of people worldwide who require treatment for HIV. In 2006, 1.3 million people had initiated ART in sub Saharan Africa, which is a 10-fold increase over the number who had access to treatment 3 years prior. Although this progress should be acknowledged, achieving universal access will require much more work. As countries initiate large-scale treatment programs, many political, social, economic, and operational challenges have become evident. South Africa, Zimbabwe, and Botswana are three neighboring countries engaged in ART roll-out. This article describes the HIV epidemic in these three countries, details the most critical challenges inhibiting the progression of antiretroviral therapy roll-out, and highlights successes within each setting. PMID- 18510894 TI - Management of hepatitis B virus co-infection on and off antiretroviral therapy. AB - Chronic hepatitis B virus (HBV) infection is recognized in 5% to 10% of persons with HIV. Co-infected individuals show an accelerated course of HBV-associated liver disease with faster progression to cirrhosis. The number of anti-HBV drugs has increased in the past few years, and some agents (eg, lamivudine, emtricitabine, tenofovir) also exert activity against HIV-1. Emergence of drug resistance challenges the long-term benefit of anti-HBV monotherapy. Data derived from studies using new more potent anti-HBV drugs are very promising, and strategies to use these antiretrovirals sequentially or in combination are being developed. Appropriate diagnosis and monitoring of chronic hepatitis B, including the use of noninvasive tools for assessing liver fibrosis, measurement of serum HBV-DNA, and drug-resistance testing, along with wise use of antivirals may convert HBV/HIV co-infection in to a manageable disease. Hopefully, this success will translate into a halt of liver-related complications and death in the co infected population. PMID- 18510897 TI - Mobility changes of the first ray after hallux valgus surgery: clinical results after proximal metatarsal chevron osteotomy and distal soft tissue procedure. AB - BACKGROUND: The purpose of this study was to evaluate the change of the first ray mobility after PMCO and DSTP in hallux valgus patients. MATERIALS AND METHODS: From May 2004 to December 2005, 82 PMCO with DSTP surgeries were performed for the management of hallux valgus deformity. The dorsiflexion mobility of the first ray of the foot was measured both preoperatively and 1 year after surgery using a modified Klaue device. The data were statistically analyzed with a paired t-test. An American Orthopedic Foot and Ankle Society (AOFAS) forefoot hallux score and patient satisfaction were also evaluated. RESULTS: Subjects consisted of 9 male and 73 female patients with an average age of 47.7 years (range, 19 to 74 years). The mean preoperative dorsiflexion mobility was 6.8 (range, 2.32 to 15.02) mm and the mean dorsiflexion mobility at one year after operation was 3.2 (range, from 1.7 to 5.4) mm. This decrease was statistically significant (p < 0.01). The mean preoperative AOFAS forefoot hallux score was 66.2 (range, 44 to 90) and improved to 89.1 (range, 72 to 100) by the 1-year followup (p < 0.01). CONCLUSION: Clinically, the dorsiflexion mobility of the first ray was significantly reduced after correction of hallux valgus with PMCO with DSTP. Because the stability of the first ray can be improved with PMCO with DSTP, the surgical indication for this procedure could include some patients showing hypermobility of the first ray. PMID- 18510898 TI - Clinical and radiological results after modified distal metatarsal osteotomy for hallux valgus. AB - BACKGROUND: The chevron osteotomy is an acceptable method for correction of mild and moderate hallux valgus, but can result in instability at the osteotomy site. The purpose of this study was to present clinical and radiological results with our modified technique of osteotomy. MATERIALS AND METHODS: We performed a modified technique of distal osteotomy of the first metatarsal on 77 feet of 46 patients with symptomatic hallux valgus; followed up for an average of 52 months. RESULTS: All of the patients experienced satisfactory pain relief and acceptable cosmesis. The mean postoperative reduction in the intermetatarsal angle was 6.5 degrees and of the metatarsophalangeal angle was 23.0 degrees. There was no loss of correction and there was no discrepancy in preoperative and postoperative lengths of the first metatarsal during the followup period. CONCLUSION: We found our modified distal metatarsal osteotomy to be an effective method of correcting hallux valgus. PMID- 18510899 TI - Modified chevron osteotomy fixed with stofella pin for hallux valgus. AB - BACKGROUND: The purpose of this study was to present the mid-term results of hallux valgus patients who underwent a modified chevron osteotomy. MATERIALS AND METHODS: Fifty-six patients (73 feet) with mild to moderate hallux valgus underwent a modified chevron osteotomy and Stoffella pin fixation between January 1999 and December 2004. Patients were evaluated clinically by the American Orthopedic Foot and Ankle Society (AOFAS) score. Pre- and postoperative radiographs were evaluated for the hallux valgus and intermetatarsal angles and sesamoid position. RESULTS: An improvement of 44.8 points in the AOFAS score was found. A change of 17.4 degrees in the hallux valgus angle and by 5.3 degrees in the intermetatarsal angle was achieved (p < 0.05). The change in the sesamoid position was significantly improved. Superficial skin infection in 3 cases, transient hypoesthesia in 2 cases, and bursitis due to screw irritation in 4 cases were the complications. CONCLUSION: Stable and rigid fixation by modified chevron osteotomy using Stoffella pins allows early mobilization and weightbearing without a cast. We believe early mobilization of the joint provides better functional outcomes with fewer complications compared to other fixation techniques. PMID- 18510900 TI - Effectiveness of ultrasound-guided corticosteroid injection in the treatment of Morton's neuroma. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy of corticosteroid injection and determine the duration of symptom-free period after treatment with a single ultrasound-guided injection for a painful Morton's neuroma. MATERIALS AND METHODS: From May 2002 to November 2003, 35 consecutive patients (7 males, 28 females) (mean age, 54; age range, 29 to 77 years) underwent a single ultrasound guided corticosteroid injection. Thirty-nine injections were performed as 4 patients had bilateral Morton's neuromas. The injection of 1.0 cc Celestone Chronodose (5.7 mg/ml) with 0.5 cc of 1% lidocaine was performed into the symptomatic intermetatarsal web-space. The efficacy of the injection was determined by the Johnson grading scale, and modified lower extremity functional scale. RESULTS: On the Johnson scale, 15 of 39 (38%) neuromas showed complete satisfaction 9 months after treatment and 11 of 39 (28%) were satisfied with minor reservations. A total of 26 of 39 (66%) neuromas had a positive outcome 9 months after the injection. On the functional daily activity (FDA) scale, 20 of 39 (51%) neuromas showed no difficulty and 4 of 39 (10%) indicated minor difficulties, which was considered a positive outcome 9 months after injection. Complete pain relief was achieved in 11 of 39 (28%) neuromas 9 months after treatment. Twelve of 39 (31%) neuromas did not respond to conservative treatment and required surgery. The results of treatment suggested improvement in efficacy if injection was used early. The size of the lesion measured on ultrasound showed no correlation with pain relief after injection. CONCLUSION: A single corticosteroid injection can offer short-term pain relief in the conservative management of Morton's neuroma. PMID- 18510901 TI - Interpositional arthroplasty with extensor digitorum brevis tendon in Freiberg's disease: a new surgical technique. AB - BACKGROUND: Freiberg's infraction is an osteochondrosis of a lesser metatarsal head resulting in joint degeneration. There is no consensus regarding the management of these lesions. Here, we describe an interpositional arthroplasty using extensor digitorum brevis tendon as a solution for Freiberg's disease. MATERIALS AND METHODS: Between 2003 and 2006, 6 women and 4 men with Freiberg's disease unresponsive to conservative treatment were operated with interpositional arthroplasty with extensor digitorum brevis tendon. Mean age was 34 (range, 20 to 48) years and followup time 24.6 (range, 12 to 36) months. The transferred tendon was passed through a tunnel, centered, stabilized and rolled into a ball following the debridement of joint. According to the Smillie classification, there were 3 grade II, 5 grade III, and 2 grade IV. The AOFAS scoring system was used for clinical assesment. RESULTS: The mean preoperative and postoperative AOFAS scores were 58.3 (range, 44 to 77) and 80.4 (range, 67 to 100), respectively. The complaint of pain with joint motion was decreased in all patients except one. The postoperative passive range of motion of joints did not differ significantly. We found 4 excellent (40%), 5 good (50%) and 1 poor (10%) result. CONCLUSION: We recommend our technique of interpositional arthroplasty with the extensor digitorum brevis tendon because it is free of additional donor site morbidity. It can be performed easily without specialized instruments. Also, the use of natural tissue eliminates potential foreign body reactions and risk of infection. PMID- 18510902 TI - Modified resection arthroplasty for infected non-healing ulcers with toe deformity in diabetic patients. AB - BACKGROUND: Diabetic motor neuropathy is expressed as the loss of function and the contracture of the intrinsic muscles of the foot, leading to the classic claw toe deformity. This deformity predisposes the foot to ulcerations on the dorsum or tip of the toes or an interdigital ulcer over a condyle between the toes. We present our results of a modified resection arthroplasty for the treatment of this difficult problem. MATERIALS AND METHODS: In this study, 72 toes (57 feet) with a deformity in the second to fifth toe accompanied by chronically infected ulcers were involved. All patients underwent modified resection arthroplasty of the PIP or DIP joint depending on the ulcer location. The second toe was involved in 27 cases (38%), the third toe in 11 cases (15%), the fourth toe in 19 cases (26%), and the fifth toe in 15 cases (21%). With the exception of 4 patients, all had a positive culture, including 7 cases of MRSA. The mean followup was 28.7 +/- 8.1 months. RESULTS: The mean wound healing time was 25.6 +/- 6.2 days. Three cases eventually required toe amputation but there was no proximal spread of infection. No recurrence of a claw toe or ulcer occurred in the remaining toes. CONCLUSION: We believe that modified resection arthroplasty for toe deformities with chronic infected ulcers in diabetic patients is a good treatment alternative to toe amputation. PMID- 18510903 TI - Achilles tendinopathy in diabetes mellitus. AB - BACKGROUND: Plantar forefoot ulcers in individuals with diabetes often lead to deep infection and lower extremity amputation. Increasing evidence suggests that the process is initiated by increased passive stiffness within the gastrocnemius soleus musculotendinous unit. The goal of this investigation was to perform ultrasound examination of the Achilles tendon in a consecutive group of asymptomatic diabetic individuals to identify any inherent structural pathology that might be associated with the increased stiffness that appears to be associated with the development of diabetic forefoot ulcers. MATERIALS AND METHODS: Seventy consecutive diabetic individuals with no past history of diabetic foot morbidity underwent ultrasonography of their Achilles tendons. Each patient was also tested for fasting glucose and glycosylated hemoglobin as a measure of diabetes control. Ten similarly aged non-diabetics with no history of Achilles tendinopathy served as controls. RESULTS: The ultrasonography revealed disorganized tendon fibers in 62 of 70 (89%) patients, and calcification within the Achilles tendon in 53 (76%) patients. The Achilles tendon thickness averaged 5.0 (range, 4 to 8) mm. There was no correlation between patient age and Achilles tendon thickness (r = 0.292, p = 0.014); however, there was a trend for duration of disease and Achilles tendon disorganization (p = 0.073). The oldest patients also appeared to demonstrate a trend for more ultrasound-measured Achilles tendinopathy. There was no correlation between fasting glucose or glycosylated hemoglobin and Achilles tendon alterations. CONCLUSION: This investigation confirms structural abnormalities within the Achilles tendon of diabetic individuals that might represent biologic changes affecting the inherent stiffness that leads to increased forefoot pressure and the development of plantar forefoot ulcers. This process appears to worsen with advanced age and does not appear to be related to diabetes control. PMID- 18510904 TI - Axial rotation and mediolateral translation of the fibula during passive plantarflexion. AB - BACKGROUND: Since the fibula is linked to the ankle as well as the knee joint, its importance for knee and ankle disabilities should be investigated. This study evaluates its movement during range of motion of the ankle. MATERIALS AND METHODS: An instrument, together with the experimental protocol, was devised to determine the relative motion of the fibula in reference to the tibia with motion of the ankle joint on 20 paired lower extremity cadaver specimens. RESULTS: It was demonstrated in all specimens that the fibula had a relative rotation around its longitudinal axis and mediolateral translation with reference to the tibia with ankle motion. The distal end of the fibula rotates more compared to the proximal end. The mediolateral translation of the proximal end of the fibula is rather close to that of the distal end. Although there was no consistent pattern for rotation, dorsiflexion caused lateral translation and plantarflexion caused medial displacement for most of the specimens. CONCLUSION: A novel, invasive but relatively simple test setup was devised. Movement of the fibula which is important for the kinematics and kinetics of the knee and ankle joints was evaluated by this new device. CLINICAL RELEVANCE: Evaluation of the fibula movement in normal lower extremities may lead to better understanding of its dynamic function which could have treatment implications for pathological conditions. PMID- 18510905 TI - Morphometric dimensions of the dorsal calcaneocuboid ligament. AB - BACKGROUND: An isolated injury to the calcaneocuboid joint is frequently unrecognized clinically. The possibility of an injured dorsal calcaneocuboid ligament (DCC) should be considered in all acutely swollen feet after trauma involving supination. Despite its significance, there is much confusion in the literature regarding nomenclature, morphology, and location. MATERIALS AND METHODS: To determine morphometric dimensions, dissection was performed on the feet of 30 adult preserved cadaveric limbs. RESULTS: The DCC was consistently present in all specimens in the form of either a solitary structure (66.6%) or 2 components (33.3%). For the solitary structures, the superior and inferior borders measured 18.6 +/- 2.8 mm and 17.3 +/- 2.5 mm, respectively. The widths of the calcaneal and cuboid sides were 13.8 +/- 2.2 mm and 12.4 +/- 2.1 mm, respectively. In the 2 component ligaments, the superior component had a superior and inferior length that averaged 16.9 +/- 2.1 mm and 16.4 +/- 2.8 mm, respectively. Its calcaneal and cuboid borders were 9.3 +/- 1.2 mm and 10.6 +/- 1.4 mm, respectively. The inferior component had superior and inferior sides that were 12.9 +/- 2.9 mm and 13.8 +/- 3.0 mm, respectively. The widths of the calcaneal and cuboid sides were 8.3 +/- 1.4 mm and 7.2 +/- 1.0 mm, respectively. CONCLUSION: We have demonstrated variations in the morphology of the DCC along with its morphometric dimensions. CLINICAL RELEVANCE: Knowledge of the morphologic variations and dimensions of the DCC can be of aid when planning for surgical management of the ruptured ligament. PMID- 18510907 TI - Technique tip: Using 1.7-mm screws in a 1/3 semitubular plate to achieve additional fixation with a first MTP fusion. PMID- 18510906 TI - Biomechanical comparison of the simple running and cross-stitch epitenon sutures in achilles tendon repairs. AB - BACKGROUND: Augmenting the strength of Achilles tendon repairs may allow for earlier active rehabilitation with less risk of adhesion formation and re ruptures, leading to quicker and stronger healing. Building upon previous research that has (1) demonstrated strength gains in Achilles repairs upon addition of simple running epitenon sutures, and (2) shown the cross-stitch epitenon suture to be stronger than the simple running stitch in flexor tendons of the hand, this study compares use of these epitenon sutures in the Achilles tendon. MATERIALS AND METHODS: Ruptures were simulated in 7 matched pairs of fresh frozen human Achilles tendons and repaired with the two-tailed Krakow locking loop core technique using No. 2 nonabsorbable, braided, polyester suture. From each pair, one specimen was randomly selected to also receive the epitenon cross-stitch, the other receiving the simple running stitch. All epitenon repairs employed 4-0 nylon suture. Repaired tendons were loaded in tension to the point of failure on a Materials Testing Machine (MTS). RESULTS: Tendon repair augmented with the cross-stitch displayed a significant, 53% greater failure strength than those repaired with the simple running stitch. Increases in initial stiffness and resistance to 2-mm gap formation in the cross-stitch specimens were 3.1% and 3.6%, respectively. CONCLUSION: Gapping resistance and initial stiffness in Achilles tendon repairs were comparable between the cross-stitch and simple running stitch, but the cross-stitch significantly improved failure strength. CLINICAL RELEVANCE: Greater failure strength may translate clinically to lower rates of re-rupture and earlier mobilization following Achilles tendon repair. PMID- 18510908 TI - Technique tip: a simplified method for washer removal. PMID- 18510909 TI - Irreducible isolated subtalar dislocation: a case report. AB - Subtalar dislocations are uncommon and account for approximately 1% of all dislocations. Optimal management is by immediate closed reduction under general anesthesia. We report 3 cases of irreducible, isolated subtalar dislocation that required an open procedure. Closed reduction failed in 2 patients with lateral dislocation due to interposition of the posterior tibialis tendon caused by a large tear of the flexor retinaculum. The flexor retinaculum was accurately reconstructed after the reduction. In the third case, a medial dislocation, a displaced extensor retinaculum prevented relocation of the talar head and required resection. We also discuss the mechanisms for irreducible subtalar dislocations. PMID- 18510910 TI - Pigmented villonodular synovitis about the ankle: two case reports. PMID- 18510911 TI - Giant cell tumor of the EDL tendon sheath: an unusual cause of hallux valgus. AB - Hallux valgus is a lateral deviation of the proximal phalanx of the first metatarsophalangeal joint. It is a common disorder in adults. The etiologic factors include modern shoes, rheumatoid arthritis, pes planus, metatarsus primus varus, and trauma. Tumors causing hallux valgus deformities are unusual. We report a 50-year-old female with a hallux valgus deformity caused by a giant cell tumor of the second EDL tendon sheath. Surgical excision of the tumor and corrective osteotomy produced a permanent cure. This unusual cause of a hallux valgus deformity should increase awareness of tumors as a possible cause of foot deformities. PMID- 18510912 TI - Mycobacterium fortuitum infection following primary achilles tendon debridement with flexor hallucis longus augmentation: a case report. AB - Mycobacterium fortuitum (M. fortuitum), a rapidly growing non-tuberculous mycobacterium is a well-recognized, yet uncommon cause of soft tissue infection. The incidence of post surgical wound infections from this organism is increasing. The presentation of infection is atypical and failure to consider this pathogen can cause diagnostic delay and increased morbidity. Achilles tendon debridement with FHL augmentation is commonly used in patients with chronic Achilles tendinosis. Wound-edge necrosis is the most common surgical complication of this procedure, and superficial and deep infections are potentially devastating complications. We report the case of a patient who underwent Achilles tendon debridement with flexor hallucis longus augmentation, whose postoperative course was complicated by a deep M. FORTUITUM infection. Critical to the identification and ultimate treatment of this particular pathogen is the utilization of appropriate intraoperative cultures and microbiologic testing. In addition, repeat aggressive irrigation and debridement procedures coupled with removal of foreign materials and the appropriate use of prolonged antibiotic therapy can result in a successful long-term outcome. PMID- 18510913 TI - Surgical strategies: insertional achilles tendinopathy. PMID- 18510914 TI - Re: syndesmosis procedure: a non-osteotomy approach to metatarsus primus varus correction. PMID- 18510915 TI - Getting up close and personal with your genome. AB - A new type of company is offering to scan a person's genome and reveal the information it holds for as little as $1000. Are these services fun novelty items or do they provide valuable information that will help people take better care of their health? PMID- 18510916 TI - Science on the streets of the Big Apple. AB - A five-day festival of science takes place this week at venues across New York City. The festival features not only leading researchers from New York and beyond but also actors, writers, musicians, and choreographers in a series of multimedia programs designed to reveal science to the general public in exciting new ways. PMID- 18510917 TI - Insect odorant receptors: channeling scent. AB - Odorant detection in insects involves heterodimers between an odorant receptor (OR) and a conserved seven-transmembrane protein called Or83b, but the exact mechanism of OR signal transduction is unclear. Two recent studies in Nature (Sato et al., 2008; Wicher et al., 2008) now reveal that these OR-Or83b heterodimers form odorant-gated ion channels, revealing a surprising new mode of olfactory transduction. PMID- 18510918 TI - Chromatin proteins do double duty. AB - The histone acetyltransferase MOF (males-absent-on-the-first) is required for the regulation of X chromosome gene dosage compensation in Drosophila males. In this issue, Kind et al. (2008) show that MOF is also found on autosomes and that it has two modes of binding: one in males for X chromosome dosage compensation and the other in both sexes for X chromosome and autosomal gene regulation independent of dosage compensation. PMID- 18510919 TI - Follow the monomer. AB - Capping proteins limit actin filament growth, but paradoxically increase actin based cell motility. This has been attributed to funneling of actin monomers to the filament ends that remain uncapped. Using a reconstituted motility system, Akin and Mullins (2008) now demonstrate that filament capping increases Arp2/3 based nucleation and branching, rather than elevating the rate of filament elongation. PMID- 18510920 TI - p53 regulation orchestrates the TGF-beta response. AB - Among its multiple functions, p53 is a critical regulator of TGF-beta responses. Sasai et al. (2008) now identify a new p53 inhibitory protein, XFDL156. During embryonic development, this factor is expressed in the ectoderm germ layer and maintains the pluripotency of ectodermal cells by inhibiting TGF-beta target genes that promote mesoderm specification. PMID- 18510921 TI - Forever young: death-defying neuroblasts. AB - During development, many neural stem cells "age" as they sequentially generate distinct neuronal or glial cell types. In this issue, Maurange et al. (2008) now identify the temporal control factors in Drosophila neural stem cells (neuroblasts) that regulate the fate of stem cell progeny and signal the end of stem cell proliferation. PMID- 18510922 TI - Plant evolution: TALES of development. AB - TALE homeodomain proteins regulate development in many eukaryotes. Now, Lee et al. (2008) report that two TALE homeodomain proteins control zygote development of the unicellular green alga Chlamydomonas. This implicates TALE gene loss and diversification in the evolution of new diploid body plans that appeared when land plants evolved from algal ancestors over 450 million years ago. PMID- 18510923 TI - Regulatory T cells and immune tolerance. AB - Regulatory T cells (Tregs) play an indispensable role in maintaining immunological unresponsiveness to self-antigens and in suppressing excessive immune responses deleterious to the host. Tregs are produced in the thymus as a functionally mature subpopulation of T cells and can also be induced from naive T cells in the periphery. Recent research reveals the cellular and molecular basis of Treg development and function and implicates dysregulation of Tregs in immunological disease. PMID- 18510924 TI - XPD helicase structures and activities: insights into the cancer and aging phenotypes from XPD mutations. AB - Mutations in XPD helicase, required for nucleotide excision repair (NER) as part of the transcription/repair complex TFIIH, cause three distinct phenotypes: cancer-prone xeroderma pigmentosum (XP), or aging disorders Cockayne syndrome (CS), and trichothiodystrophy (TTD). To clarify molecular differences underlying these diseases, we determined crystal structures of the XPD catalytic core from Sulfolobus acidocaldarius and measured mutant enzyme activities. Substrate binding grooves separate adjacent Rad51/RecA-like helicase domains (HD1, HD2) and an arch formed by 4FeS and Arch domains. XP mutations map along the HD1 ATP binding edge and HD2 DNA-binding channel and impair helicase activity essential for NER. XP/CS mutations both impair helicase activity and likely affect HD2 functional movement. TTD mutants lose or retain helicase activity but map to sites in all four domains expected to cause framework defects impacting TFIIH integrity. These results provide a foundation for understanding disease consequences of mutations in XPD and related 4Fe-4S helicases including FancJ. PMID- 18510925 TI - Structure of the DNA repair helicase XPD. AB - The XPD helicase (Rad3 in Saccharomyces cerevisiae) is a component of transcription factor IIH (TFIIH), which functions in transcription initiation and Nucleotide Excision Repair in eukaryotes, catalyzing DNA duplex opening localized to the transcription start site or site of DNA damage, respectively. XPD has a 5' to 3' polarity and the helicase activity is dependent on an iron-sulfur cluster binding domain, a feature that is conserved in related helicases such as FancJ. The xpd gene is the target of mutation in patients with xeroderma pigmentosum, trichothiodystrophy, and Cockayne's syndrome, characterized by a wide spectrum of symptoms ranging from cancer susceptibility to neurological and developmental defects. The 2.25 A crystal structure of XPD from the crenarchaeon Sulfolobus tokodaii, presented here together with detailed biochemical analyses, allows a molecular understanding of the structural basis for helicase activity and explains the phenotypes of xpd mutations in humans. PMID- 18510926 TI - Genome-wide analysis reveals MOF as a key regulator of dosage compensation and gene expression in Drosophila. AB - Dosage compensation, mediated by the MSL complex, regulates X-chromosomal gene expression in Drosophila. Here we report that the histone H4 lysine 16 (H4K16) specific histone acetyltransferase MOF displays differential binding behavior depending on whether the target gene is located on the X chromosome versus the autosomes. More specifically, on the male X chromosome, where MSL1 and MSL3 are preferentially associated with the 3' end of dosage compensated genes, MOF displays a bimodal distribution binding to promoters and the 3' ends of genes. In contrast, on MSL1/MSL3 independent X-linked genes and autosomal genes in males and females, MOF binds primarily to promoters. Binding of MOF to autosomes is functional, as H4K16 acetylation and the transcription levels of a number of genes are affected upon MOF depletion. Therefore, MOF is not only involved in the onset of dosage compensation, but also acts as a regulator of gene expression in the Drosophila genome. PMID- 18510927 TI - Early sexual origins of homeoprotein heterodimerization and evolution of the plant KNOX/BELL family. AB - Developmental mechanisms that yield multicellular diversity are proving to be well conserved within lineages, generating interest in their origins in unicellular ancestors. We report that molecular regulation of the haploid-diploid transition in Chlamydomonas, a unicellular green soil alga, shares common ancestry with differentiation pathways in land plants. Two homeoproteins, Gsp1 and Gsm1, contributed by gametes of plus and minus mating types respectively, physically interact and translocate from the cytosol to the nucleus upon gametic fusion, initiating zygote development. Their ectopic expression activates zygote development in vegetative cells and, in a diploid background, the resulting zygotes undergo a normal meiosis. Gsm1/Gsp1 dyads share sequence homology with and are functionally related to KNOX/BELL dyads regulating stem-cell (meristem) specification in land plants. We propose that combinatorial homeoprotein-based transcriptional control, a core feature of the fungal/animal radiation, may have originated in a sexual context and enabled the evolution of land-plant body plans. PMID- 18510928 TI - Capping protein increases the rate of actin-based motility by promoting filament nucleation by the Arp2/3 complex. AB - Capping protein (CP) is an integral component of Arp2/3-nucleated actin networks that drive amoeboid motility. Increasing the concentration of capping protein, which caps barbed ends of actin filaments and prevents elongation, increases the rate of actin-based motility in vivo and in vitro. We studied the synergy between CP and Arp2/3 using an in vitro actin-based motility system reconstituted from purified proteins. We find that capping protein increases the rate of motility by promoting more frequent filament nucleation by the Arp2/3 complex and not by increasing the rate of filament elongation as previously suggested. One consequence of this coupling between capping and nucleation is that, while the rate of motility depends strongly on the concentration of CP and Arp2/3, the net rate of actin assembly is insensitive to changes in either factor. By reorganizing their architecture, dendritic actin networks harness the same assembly kinetics to drive different rates of motility. PMID- 18510929 TI - The big brain aquaporin is required for endosome maturation and notch receptor trafficking. AB - Activity of the big brain (bib) gene influences Notch signaling during Drosophila nervous system development. We demonstrate that Bib, which belongs to the aquaporin family of channel proteins, is required for endosome maturation in Drosophila epithelial cells. In the absence of Bib, early endosomes arrest and form abnormal clusters, and cells exhibit reduced acidification of endocytic trafficking organelles. Bib acts downstream of Hrs in early endosome morphogenesis and regulates biogenesis of endocytic compartments prior to the formation of Rab7-containing late endosomes. Abnormal endosome morphology caused by loss of Bib is accompanied by overaccumulation of Notch, Delta, and other signaling molecules as well as reduced intracellular trafficking of Notch to nuclei. Analysis of several endosomal trafficking mutants reveals a correlation between endosomal acidification and levels of Notch signaling. Our findings reveal an unprecedented role for an aquaporin in endosome maturation, trafficking, and acidification. PMID- 18510930 TI - Chk1 suppresses a caspase-2 apoptotic response to DNA damage that bypasses p53, Bcl-2, and caspase-3. AB - Evasion of DNA damage-induced cell death, via mutation of the p53 tumor suppressor or overexpression of prosurvival Bcl-2 family proteins, is a key step toward malignant transformation and therapeutic resistance. We report that depletion or acute inhibition of checkpoint kinase 1 (Chk1) is sufficient to restore gamma-radiation-induced apoptosis in p53 mutant zebrafish embryos. Surprisingly, caspase-3 is not activated prior to DNA fragmentation, in contrast to classical intrinsic or extrinsic apoptosis. Rather, an alternative apoptotic program is engaged that cell autonomously requires atm (ataxia telangiectasia mutated), atr (ATM and Rad3-related) and caspase-2, and is not affected by p53 loss or overexpression of bcl-2/xl. Similarly, Chk1 inhibitor-treated human tumor cells hyperactivate ATM, ATR, and caspase-2 after gamma-radiation and trigger a caspase-2-dependent apoptotic program that bypasses p53 deficiency and excess Bcl 2. The evolutionarily conserved "Chk1-suppressed" pathway defines a novel apoptotic process, whose responsiveness to Chk1 inhibitors and insensitivity to p53 and BCL2 alterations have important implications for cancer therapy. PMID- 18510931 TI - Ectodermal factor restricts mesoderm differentiation by inhibiting p53. AB - During gastrulation of the amphibian embryo, specification of the three germ layers, endo-, ecto-, and mesoderm, is regulated by maternal and zygotic mechanisms. Although it is known that mesoderm specification requires the cooperation between TGF-beta signaling and p53 activity and requires maternal factors, essential zygotic factors have been elusive. Here, we report that the Zn finger protein XFDL156 is an ectodermal, zygotic factor that suppresses mesodermal differentiation. XFDL156 overexpression suppresses mesodermal markers, and its depletion induces aberrant mesodermal differentiation in the presumptive ectoderm. Furthermore, we find that XFDL156 and its mammalian homologs interact with the C-terminal regulatory region of p53, thereby inhibiting p53 target gene induction and mesodermal differentiation. Thus, XFDL156 actively restricts mesodermal differentiation in the presumptive ectoderm by controlling the spatiotemporal responsiveness to p53. PMID- 18510932 TI - Temporal transcription factors and their targets schedule the end of neural proliferation in Drosophila. AB - The timing mechanisms responsible for terminating cell proliferation toward the end of development remain unclear. In the Drosophila CNS, individual progenitors called neuroblasts are known to express a series of transcription factors endowing daughter neurons with different temporal identities. Here we show that Castor and Seven-Up, members of this temporal series, regulate key events in many different neuroblast lineages during late neurogenesis. First, they schedule a switch in the cell size and identity of neurons involving the targets Chinmo and Broad Complex. Second, they regulate the time at which neuroblasts undergo Prospero-dependent cell-cycle exit or Reaper/Hid/Grim-dependent apoptosis. Both types of progenitor termination require the combined action of a late phase of the temporal series and indirect feedforward via Castor targets such as Grainyhead and Dichaete. These studies identify the timing mechanism ending CNS proliferation and reveal how aging progenitors transduce bursts of transcription factors into long-lasting changes in cell proliferation and cell identity. PMID- 18510933 TI - The microRNA miR-1 regulates a MEF-2-dependent retrograde signal at neuromuscular junctions. AB - We show that miR-1, a conserved muscle-specific microRNA, regulates aspects of both pre- and postsynaptic function at C. elegans neuromuscular junctions. miR-1 regulates the expression level of two nicotinic acetylcholine receptor (nAChR) subunits (UNC-29 and UNC-63), thereby altering muscle sensitivity to acetylcholine (ACh). miR-1 also regulates the muscle transcription factor MEF-2, which results in altered presynaptic ACh secretion, suggesting that MEF-2 activity in muscles controls a retrograde signal. The effect of the MEF-2 dependent retrograde signal on secretion is mediated by the synaptic vesicle protein RAB-3. Finally, acute activation of levamisole-sensitive nAChRs stimulates MEF-2-dependent transcriptional responses and induces the MEF-2 dependent retrograde signal. We propose that miR-1 refines synaptic function by coupling changes in muscle activity to changes in presynaptic function. PMID- 18510934 TI - Live imaging of neuronal degradation by microglia reveals a role for v0-ATPase a1 in phagosomal fusion in vivo. AB - A significant proportion of neurons in the brain undergo programmed cell death. In order to prevent the diffusion of damaging degradation products, dying neurons are quickly digested by microglia. Despite the importance of microglia in several neuronal pathologies, the mechanism underlying their degradation of neurons remains elusive. Here, we exploit a microglial population in the zebrafish to study this process in intact living brains. In vivo imaging reveals that digestion of neurons occurs in compartments arising from the progressive fusion of vesicles. We demonstrate that this fusion is mediated by the v0-ATPase a1 subunit. By applying live pH indicators, we show that the a1 subunit mediates fusion between phagosomes and lysosomes during phagocytosis, a function that is independent of its proton pump activity. As a real-time description of microglial phagocytosis in vivo, this work advances our understanding of microglial-mediated neuronal degeneration, a hallmark of many neuronal diseases. PMID- 18510935 TI - SnapShot: p53 posttranslational modifications. PMID- 18510936 TI - Amine-reactive isobaric tagging reagents: requirements for absolute quantification of proteins and peptides. AB - Amine-reactive isobaric tagging reagents such as iTRAQ (isobaric tags for relative and absolute quantitation) have recently become increasing popular for relative protein quantification, cell expression profiling, and biomarker discovery. This is due mainly to the possibility of simultaneously identifying and quantifying multiple samples. The principles of iTRAQ may also be applied to absolute protein quantification with the use of synthetic peptides as standards. The prerequisites that must be fulfilled to perform absolute quantification of proteins by iTRAQ have been investigated and are described here. Three samples of somatropin were quantified using iTRAQ and synthetic peptides as standards, corresponding to a portion of the protein sequence. The results were compared with those obtained by quantification of the same protein solutions using double exact matching isotope dilution mass spectrometry (IDMS). To obtain reliable results, the appropriate standard peptides needed to be selected carefully and enzymatic digestion needed to be optimized to ensure complete release of the peptides from the protein. The kinetics and efficiency of the iTRAQ derivatization reaction of the standard peptides and digested proteins with isobaric tagging reagents were studied using a mixture of seven synthetic peptides and their corresponding labeled peptides. The implications of incomplete derivatization are also presented. PMID- 18510937 TI - Plant protein isolation and stabilization for enhanced resolution of two dimensional polyacrylamide gel electrophoresis. AB - Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is the common method of choice for proteomic analysis. By introducing several small changes, a method was developed that not only improved the resolution and reproducibility of 2D PAGE but also shortened the time of analysis. Precipitation by alkaline phenol and methanol/ammonium acetate was the choice for protein extraction. However, instead of precipitating the proteins overnight at -20 degrees C, it was carried out for 2 to 3h at -80 degrees C. Ethanol was used for the final wash of the protein precipitate instead of routinely used acetone. Dithiothreitol (DTT) was used in all solutions from the beginning, considerably improving the solubilization of precipitated proteins. Solubilization was further improved by using a mixture of detergents and denaturants at high concentrations along with large amounts of DTT. Both in-gel rehydration and cup-loading methods were used for isoelectric focusing (IEF). For in-gel rehydration, samples reduced with DTT were diluted with sample buffer containing 2-hydroxyethyl disulfide (2-HED) (1:3) or were cup-loaded on a strip rehydrated with sample buffer containing 2-HED. Glycerol (5%) was used in the sample buffer, and the focusing was performed at 15 degrees C. The applicability of the method was demonstrated using several soybean tissues. PMID- 18510938 TI - A critical review and discussion of analytical methods in the L-arginine/nitric oxide area of basic and clinical research. PMID- 18510939 TI - E1AF promotes mithramycin A-induced Huh-7 cell apoptosis depending on its DNA binding domain. AB - Transcription factor E1AF is widely known to play critical roles in tumor metastasis via directly binding to the promoters of genes involved in tumor migration and invasion. Here, we reported for the first time the pro-apoptotic role of E1AF in tumor cells. The expression of E1AF at protein level was obviously increased during Huh-7 and Hep3B cells apoptosis induced by the anticancer agent mithramycin A. E1AF overexpression markedly enhanced mithramycin A-induced Huh-7 cell apoptosis and the expression of pro-apoptotic protein Bax depending on its DNA-binding domain. And, reduction of E1AF inhibited mithramycin A-induced Huh-7 cell apoptosis. Furthermore, reducing the expression of Bax significantly inhibited E1AF-increased Huh-7 cell apoptosis induced by mithramycin A. Taken together, E1AF increases mithramycin A-induced Huh-7 cells apoptosis and Bax expression depending on its DNA-binding domain, indicating that E1AF might contribute to the therapeutic efficiency of mithramycin A for hepatoma. PMID- 18510940 TI - Correlated light and electron microscopy illuminates the role of mitochondrial inner membrane remodeling during apoptosis. AB - In addition to their role in providing ATP for cellular functions via oxidative phosphorylation, mitochondria also play a critical role in initiating and/or regulating apoptosis through the release of proteins such as cytochrome c from intermembrane and intracristal compartments. The mechanism by which these proteins are able to cross the outer mitochondrial membrane has been a subject of controversy. This paper will review some recent results that demonstrate that inner mitochondrial membrane remodeling does occur during apoptosis in HeLa cells but does not appear to be a requirement for release of cytochrome c from intracristal compartments. Inner membrane remodeling does appear to be related to fragmentation of the mitochondrial matrix, and the form of the remodeling suggests a topological mechanism for inner membrane fission and fusion. PMID- 18510941 TI - Acceptor side effects on the electron transfer at cryogenic temperatures in intact photosystem II. AB - In intact PSII, both the secondary electron donor (Tyr(Z)) and side-path electron donors (Car/Chl(Z)/Cyt(b)(559)) can be oxidized by P(680)(+)* at cryogenic temperatures. In this paper, the effects of acceptor side, especially the redox state of the non-heme iron, on the donor side electron transfer induced by visible light at cryogenic temperatures were studied by EPR spectroscopy. We found that the formation and decay of the S(1)Tyr(Z) EPR signal were independent of the treatment of K(3)Fe(CN)(6), whereas formation and decay of the Car(+)/Chl(Z)(+) EPR signal correlated with the reduction and recovery of the Fe(3+) EPR signal of the non-heme iron in K(3)Fe(CN)(6) pre-treated PSII, respectively. Based on the observed correlation between Car/Chl(Z) oxidation and Fe(3+) reduction, the oxidation of non-heme iron by K(3)Fe(CN)(6) at 0 degrees C was quantified, which showed that around 50-60% fractions of the reaction centers gave rise to the Fe(3+) EPR signal. In addition, we found that the presence of phenyl-p-benzoquinone significantly enhanced the yield of Tyr(Z) oxidation. These results indicate that the electron transfer at the donor side can be significantly modified by changes at the acceptor side, and indicate that two types of reaction centers are present in intact PSII, namely, one contains unoxidizable non-heme iron and another one contains oxidizable non-heme iron. Tyr(Z) oxidation and side-path reaction occur separately in these two types of reaction centers, instead of competition with each other in the same reaction centers. In addition, our results show that the non-heme iron has different properties in active and inactive PSII. The oxidation of non-heme iron by K(3)Fe(CN)(6) takes place only in inactive PSII, which implies that the Fe(3+) state is probably not the intermediate species for the turnover of quinone reduction. PMID- 18510942 TI - Sites of generation of reactive oxygen species in homogenates of brain tissue determined with the use of respiratory substrates and inhibitors. AB - Reactive oxygen species (ROS) have been widely implicated in the pathogenesis of various neurological diseases and aging. But the exact sites of ROS generation in brain tissue remained so far elusive. Here, we provide direct experimental evidence that at least 50% of total ROS generation in succinate-oxidizing homogenates of brain tissue can be attributed to complex I of mitochondrial respiratory chain. Applying quantitative methods for ROS detection we observed in different preparations from human, rat and mouse brain (digitonin-permeabilized tissue homogenates and isolated mitochondria) a linear relationship between rate of oxygen consumption and ROS generation with succinate as mitochondrial substrate. This quantitative relationship indicates, that under the particular conditions of oxygen saturation about 1% of the corresponding respiratory chain electron flow is redirected to form superoxide. Since we observed in mouse and rat brain mitochondria a unique dependency of both forward and reverse electron flow-dependent mitochondrial H(2)O(2) production on NAD redox state, we substantiated previous evidence that the FMN moiety of complex I is the major donor of electrons for the single electron reduction of molecular oxygen. PMID- 18510943 TI - The ADP and ATP transport in mitochondria and its carrier. AB - Different from some more specialised short reviews, here a general although not encyclopaedic survey of the function, metabolic role, structure and mechanism of the ADP/ATP transport in mitochondria is presented. The obvious need for an "old fashioned" review comes from the gateway role in metabolism of the ATP transfer to the cytosol from mitochondria. Amidst the labours, 40 or more years ago, of unravelling the role of mitochondrial compartments and of the two membranes, the sequence of steps of how ATP arrives in the cytosol became a major issue. When the dust settled, a picture emerged where ATP is exported across the inner membrane in a 1:1 exchange against ADP and where the selection of ATP versus ADP is controlled by the high membrane potential at the inner membrane, thus uplifting the free energy of ATP in the cytosol over the mitochondrial matrix. Thus the disparate energy and redox states of the two major compartments are bridged by two membrane potential responsive carriers to enable their symbiosis in the eukaryotic cell. The advance to the molecular level by studying the binding of nucleotides and inhibitors was facilitated by the high level of carrier (AAC) binding sites in the mitochondrial membrane. A striking flexibility of nucleotide binding uncovered the reorientation of carrier sites between outer and inner face, assisted by the side specific high affinity inhibitors. The evidence of a single carrier site versus separate sites for substrate and inhibitors was expounded. In an ideal setting principles of transport catalysis were elucidated. The isolation of intact AAC as a first for any transporter enabled the reconstitution of transport for unravelling, independently of mitochondrial complications, the factors controlling the ADP/ATP exchange. Electrical currents measured with the reconstituted AAC demonstrated electrogenic translocation and charge shift of reorienting carrier sites. Aberrant or vital para-functions of AAC in basal uncoupling and in the mitochondrial pore transition were demonstrated in mitochondria and by patch clamp with reconstituted AAC. The first amino acid sequence of AAC and of any eukaryotic carrier furnished a 6-transmembrane helix folding model, and was the basis for mapping the structure by access studies with various probes, and for demonstrating the strong conformation changes demanded by the reorientation mechanism. Mutations served to elucidate the function of residues, including the particular sensitivity of ATP versus ADP transport to deletion of critical positive charge in AAC. After resisting for decades, at last the atomic crystal structure of the stabilised CAT-AAC complex emerged supporting the predicted principle fold of the AAC but showing unexpected features relevant to mechanism. Being a snapshot of an extreme abortive "c-state" the actual mechanism still remains a conjecture. PMID- 18510944 TI - Cholesterol's decoupling effect on membrane partitioning and permeability revisited: is there anything beyond Fick's law of diffusion? AB - In general, Fick's law of diffusion describes membrane permeation of hydrophobic or amphiphilic molecules. In contrast to this, Thomae et al. recently identified the volume ratio between barrier and aqueous compartments as important additional determinants of membrane permeability (Pm) [A.V. Thomae, T. Koch, C. Panse, H. Wunderli-Allenspach, and S.D. Kramer, Comparing the lipid membrane affinity and permeation of drug-like acids: the intriguing effects of cholesterol and charged lipids, Pharm. Res. 24 (2007) 1457-1472.]. This new theory was supported by the striking observation that low concentrations of cholesterol increased Pm of salicylic acid. As Fick's law is of fundamental importance to all membrane transport processes, we reinvestigated this phenomenon. We measured the electrophoretic mobility of vesicles and used electrochemical scanning microscopy to study the adsorption of the SA anion to lipid vesicular bilayers and SA transport through planar lipid bilayers, respectively. As predicted by Fick's law, Pm of SA decreased continuously with increasing cholesterol content. Thomae et al. made the contrasting artifactual observation because their kinetic approach lacked the required time resolution and led to an underestimation of Pm by five orders of magnitude. We conclude that there is nothing beyond Fick's law of diffusion. It is still valid. PMID- 18510945 TI - PI3K signaling supports amphetamine-induced dopamine efflux. AB - The dopamine (DA) transporter (DAT) is a major molecular target of the psychostimulant amphetamine (AMPH). AMPH, as a result of its ability to reverse DAT-mediated inward transport of DA, induces DA efflux thereby increasing extracellular DA levels. This increase is thought to underlie the behavioral effects of AMPH. We have demonstrated previously that insulin, through phosphatidylinositol 3-kinase (PI3K) signaling, regulates DA clearance by fine tuning DAT plasma membrane expression. PI3K signaling may represent a novel mechanism for regulating DA efflux evoked by AMPH, since only active DAT at the plasma membrane can efflux DA. Here, we show in both a heterologous expression system and DA neurons that inhibition of PI3K decreases DAT cell surface expression and, as a consequence, AMPH-induced DA efflux. PMID- 18510946 TI - Physical interaction between Tbx6 and mespb is indispensable for the activation of bowline expression during Xenopus somitogenesis. AB - During vertebrate somitogenesis, various transcriptional factors function coordinately to determine the position of the somite boundary. Previously, we reported on the signaling crosstalk that occurs between two major transcription factors involved in somitogenesis, Tbx6 and mespb/mesp2. These factors synergistically activated the expression of a downstream gene, bowline/Ripply2, which is essential for precise formation of the somite boundary. However, the molecular mechanism underlying this synergistic effect remains unclear. In this report, we found that the Tbx6 and mespb proteins interacted physically with each other. Pulldown assays with various deletion mutants of these proteins identified the essential domains for this physical interaction. Finally, we found that interference with the physical interaction by a dominant-negative form of mespb, mespbDeltaDBD, abrogated the expression of the bowline gene during Xenopus somitogenesis. These results indicate that the appropriate expression of bowline/Ripply2 is regulated by a direct interaction between the Tbx6 and mespb proteins during Xenopus somitogenesis. PMID- 18510947 TI - Biliverdin is the endogenous ligand of human serum alpha1-acid glycoprotein. AB - alpha(1)-Acid glycoprotein (AAG), an acute phase component of the human serum, is a prominent member of the lipocalin family of proteins showing inflammatory/immunomodulatory activities and promiscuous drug binding properties. Both three-dimensional structure of AAG and its precise biological function are still unknown and only a few endogenous AAG ligands have been described to date. CD spectroscopic studies performed with commercial AAG and the separated genetic variants revealed high-affinity binding of biliverdin (BV) and biliverdin dimethyl ester to the 'F1/S' fraction of the protein. The preferential accommodation of the right-handed, P-helicity conformers of the pigments by the protein matrix resulted in strong induced CD activity, which was utilized for estimation of the binding parameters and to locate the binding site. It was concluded that both pigments are bound in the central beta-barrel cavity of AAG, held principally by hydrophobic interactions. Possible biological implications of the BV binding ability of AAG with special emphasis on the heme oxygenase-1 pathway are discussed. PMID- 18510948 TI - Template enhanced activity of lipase accommodated in siliceous mesocellular foams. AB - Lipases were adsorbed in siliceous mesocellular foams containing different amounts of residual template in the nanopores. It is found that the hydrolytic activities of the adsorbed lipases are increased with increasing the contents of template in the mesopores. The triacetin hydrolytic activity of the lipase adsorbed in the foam containing 46% of template can be 13 times higher than that of the lipase adsorbed in the foam without template in the nanopores, and its specific activity is about three times higher than that of the free lipase, showing the hyperactivation effect on lipase resulting from the interaction between the lipase and the surfactant in the nanopores. The immobilized lipase cross-linked with glutaraldehyde can retain up to 88% of its original activity after six hydrolysis reaction test. This work provides a new strategy to enhance the activity of immobilized lipase in mesoporous materials. PMID- 18510949 TI - MicroRNA switches in Trypanosoma brucei. AB - Trypanosoma brucei develops chronic infection in mammalian hosts due to a sophisticated strategy of antigenic variation of variant surface glycoprotein (VSG) coat to escape antibody-mediated lysis. MicroRNAs are a class of non-coding RNAs with presumed post-transcriptional regulatory activity. Homology based informatic approach is used to identify the microRNA (miRNA) genes of T. brucei and their target mRNAs. Our observation reveals a set of microRNAs targeting mRNAs corresponding to VSGs. Further, a number of miRNA hairpins have been found in clusters of multiple identical copies. The target proteins, 20S proteosome, GM6 and GRESAG 4.2 corresponding to these clustered miRNAs play essential role in trypanosomiasis. These snippets can act as genetic switches modulating host parasite interaction and provide useful clue toward treatment of trypanosomiasis. PMID- 18510950 TI - Involvement of nitric oxide in the promotion of cell survival by ceramide 1 phosphate. AB - Macrophages play vital roles in inflammatory responses, and their number at sites of inflammation is strictly regulated by cell death and division. Here, we demonstrate that production of nitric oxide (NO) is a major mechanism whereby ceramide-1-phosphate (C1P) blocks apoptosis in macrophages. However, NO failed to stimulate macrophage proliferation. The prosurvival effect of C1P was blocked by inhibitors of inducible NO synthase. The antiapoptotic effect of C1P was also blocked by phosphatidylinositol 3-kinase or nuclear factor-kappa B inhibitors. Moreover, NO reversed the inhibitory effect of C1P on acid sphingomyelinase, but the prosurvival effect of C1P was independent of this action. PMID- 18510951 TI - Words of wisdom. Re: A sealed laparoscopic nephroureterectomy: a new technique. PMID- 18510952 TI - Words of wisdom. Re: obesity-related plasma hemodilution and PSA concentration among men with prostate cancer. PMID- 18510953 TI - Words of wisdom. Re: needle biopsies on autopsy prostates: sensitivity of cancer detection based on true prevalence. PMID- 18510954 TI - Words of wisdom. Re: under diagnosis and over diagnosis of prostate cancer. PMID- 18510955 TI - Words of wisdom. Re: determining dosing intervals for LHRH agonists based on serum testosterone levels: a prospective study. PMID- 18510956 TI - Transseptal left heart catheterization a 50-year odyssey. AB - Development in the 1950s of the transseptal technique for left heart catheterization is described. Initial studies in animals and human cadavers were followed up by left atrial puncture with measurements of left atrial and left ventricular (LV) pressure (the latter using a small plastic catheter) in patients with cardiac disease. Many such procedures were performed safely without complications. Subsequent modification of the original technique for percutaneous catheter insertion allowed placement of a larger taper-tipped catheter in the LV chamber for selective LV angiography. Early clinical research studies at the National Heart Institute were performed using the transseptal method; these included investigation of the effects of increasing afterload on the normal and failing left ventricle by means of a graded angiotensin infusion to induce a progressive increase in aortic pressure. A marked decrease in the stroke volume occurred with increased afterload in the failing heart. This finding later led to the concept of afterload mismatch with limited pre-load reserve. Another early transseptal catheterization study in which measurements of LV pressure were made at different locations within the left ventricle as well as in the left atrium confirmed the presence of cavity obliteration in some patients and true obstruction in the LV outflow tract in many others. In addition, left ventriculography showed that obstruction was caused by abnormal anterior position during systole of the anterior mitral valve leaflet. With growing acceptance of retrograde catheterization of the left ventricle, the use of the transseptal technique for diagnostic purposes declined. However, in recent years, substantial renewed application of the transseptal method has occurred for special diagnostic and therapeutic purposes, including balloon valvuloplasties and electrophysiologic ablation procedures within the left heart. PMID- 18510957 TI - Emerging applications for transseptal left heart catheterization old techniques for new procedures. AB - Transseptal (TS) catheterization was introduced in 1959 as a strategy to directly measure left atrial (LA) pressure. Despite acceptable feasibility and safety, TS catheterization has been replaced by indirect measurements of LA pressure using the Swan-Ganz catheter. Today, TS puncture is rarely performed for diagnostic purposes but continues to be performed for procedures such as balloon mitral valvuloplasty, antegrade balloon aortic valvuloplasty, and ablation of arrhythmias in the LA. Thus, the "art" of TS puncture has been lost, except in centers that perform these procedures with regularity. Recently, there has been a renewed interest in the TS technique because of emerging therapeutic procedures for structural heart disease and atrial fibrillation ablation. Invasive cardiologists will have to refamiliarize themselves with the TS technique, newer TS devices, and advanced ultrasound imaging for guidance of the procedure. PMID- 18510958 TI - Differential effects of drug-eluting stents on local endothelium-dependent coronary vasomotion. AB - OBJECTIVES: The aim of our study was to compare coronary vasomotion after implantation of a second-generation biolimus A9-eluting stent (BES) and of a sirolimus-eluting stent (SES). BACKGROUND: Drug-eluting stents (DES) have been associated with impaired local coronary vasomotion, delayed endothelialization, and increased late thrombotic risk. New DES with different drugs, pharmacokinetics, and polymers have been developed. METHODS: Nineteen patients with a BES and 15 patients with a SES were studied 9 months after stent implantation. Endothelium-dependent and -independent coronary vasomotion were tested proximally and distally to the stent as well as at a reference segment during right atrial pacing at increasing heart rates. Quantitative coronary angiographic measurements were performed offline. RESULTS: Of the patients with BES, 2 showed vasoconstriction with increased heart rate and 17 showed vasodilatation. Of the patients with a SES, 9 showed vasoconstriction while 6 showed vasodilatation. The SES showed significant vasoconstriction at both the proximal (-2.3 +/- 10% vs. 7.9 +/- 10%) and the distal (-5.4 +/- 9% vs. 6.1 +/- 8%) segments to the stent compared with the BES (p = 0.003 for proximal, p < 0.001 for distal segment). Endothelium-independent vasomotion after intracoronary nitrates did not differ significantly between the 2 groups (p = NS for proximal and distal segment). CONCLUSIONS: Unlike the case with the SES, endothelium dependent vasomotion at adjacent stent segments seems to be preserved after BES implantation. This result may be explained by the different drug release kinetics, DES design, or characteristics of polymer used in the stent system. PMID- 18510959 TI - Sirolimus-induced vascular dysfunction. Increased mitochondrial and nicotinamide adenosine dinucleotide phosphate oxidase-dependent superoxide production and decreased vascular nitric oxide formation. AB - OBJECTIVES: This study sought to analyze mechanisms that mediate vascular dysfunction induced by sirolimus. BACKGROUND: Despite excellent antirestenotic capacity, sirolimus-eluting stents have been found to trigger coronary endothelial dysfunction and impaired re-endothelialization. METHODS: To mimic the continuous sirolimus exposure of a stented vessel, Wistar rats underwent drug infusion with an osmotic pump for 7 days. RESULTS: Sirolimus treatment caused a marked degree of endothelial dysfunction as well as a desensitization of the vasculature to the endothelium-independent vasodilator nitroglycerin. Also, sirolimus stimulated intense transmural superoxide formation as detected by dihydroethidine fluorescence in aortae. Increased superoxide production was mediated in part by the vascular nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase as indicated by a marked stimulation of p67(phox)/rac1 NADPH oxidase subunit expression and by increased rac1 membrane association. In addition, superoxide production in rat heart mitochondria was up-regulated by sirolimus, as measured by L012-enhanced chemiluminescence. As a consequence, electron spin resonance measurements showed a 40% reduction in vascular nitric oxide bioavailability, which was further supported by decreased serum nitrite levels. CONCLUSIONS: Sirolimus causes marked vascular dysfunction and nitrate resistance after continuous treatment for 7 days. This impaired vasorelaxation may, in part, be induced by up-regulated mitochondrial superoxide release as well as by an up-regulation of NADPH oxidase-driven superoxide production. Both processes could contribute to endothelial dysfunction observed after coronary vascular interventions with sirolimus-coated stents. PMID- 18510960 TI - Endothelial dysfunction associated with drug-eluting stents what, where, when, and how? PMID- 18510961 TI - Sustained benefits in vascular function through flavanol-containing cocoa in medicated diabetic patients a double-masked, randomized, controlled trial. AB - OBJECTIVES: Our goal was to test feasibility and efficacy of a dietary intervention based on daily intake of flavanol-containing cocoa for improving vascular function of medicated diabetic patients. BACKGROUND: Even in fully medicated diabetic patients, overall prognosis is unfavorable due to deteriorated cardiovascular function. Based on epidemiological data, diets rich in flavanols are associated with a reduced cardiovascular risk. METHODS: In a feasibility study with 10 diabetic patients, we assessed vascular function as flow-mediated dilation (FMD) of the brachial artery, plasma levels of flavanol metabolites, and tolerability after an acute, single-dose ingestion of cocoa, containing increasing concentrations of flavanols (75, 371, and 963 mg). In a subsequent efficacy study, changes in vascular function in 41 medicated diabetic patients were assessed after a 30-day, thrice-daily dietary intervention with either flavanol-rich cocoa (321 mg flavanols per dose) or a nutrient-matched control (25 mg flavanols per dose). Both studies were undertaken in a randomized, double masked fashion. Primary and secondary outcome measures included changes in FMD and plasma flavanol metabolites, respectively. RESULTS: A single ingestion of flavanol-containing cocoa was dose-dependently associated with significant acute increases in circulating flavanols and FMD (at 2 h: from 3.7 +/- 0.2% to 5.5 +/- 0.4%, p < 0.001). A 30-day, thrice-daily consumption of flavanol-containing cocoa increased baseline FMD by 30% (p < 0.0001), while acute increases of FMD upon ingestion of flavanol-containing cocoa continued to be manifest throughout the study. Treatment was well tolerated without evidence of tachyphylaxia. Endothelium-independent responses, blood pressure, heart rate, and glycemic control were unaffected. CONCLUSIONS: Diets rich in flavanols reverse vascular dysfunction in diabetes, highlighting therapeutic potentials in cardiovascular disease. PMID- 18510962 TI - Flavanol-rich cocoa a promising new dietary intervention to reduce cardiovascular risk in type 2 diabetes? PMID- 18510963 TI - Atrial fibrillation begets atrial fibrillation in the pulmonary veins on the impact of atrial fibrillation on the electrophysiological properties of the pulmonary veins in humans. AB - OBJECTIVES: Our purpose was to investigate the impact of short-lasting atrial fibrillation (AF) on the electrophysiological properties of the atria and pulmonary veins (PVs) in patients devoid of AF. BACKGROUND: The presence of AF is associated with electrical remodeling processes that promote a substrate for arrhythmia maintenance in the atria, which has been termed "AF begets AF." However, it is unclear whether those electrical alterations also occur in the PVs. METHODS: Thirty-five patients with a left-sided accessory pathway and without a prior history of AF were included. After successful ablation, the effective refractory periods (ERPs) and conduction times of the right atrium (RA), left atrium (LA), and the PVs were determined. Afterwards, AF was induced and maintained for a period of 15 min. Thereafter, the stimulation protocol was repeated. RESULTS: At baseline, the PVs had significantly longer ERPs than the atria. After exposure to AF, the ERPs of both the atria and the PVs decreased significantly. The ERPs of the PVs, however, decreased by a significantly greater extent than the ERPs of the atria (PVs: 248 +/- 27 ms vs. 211 +/- 40 ms, p < 0.001; LA: 233 +/- 23 ms vs. 214 +/- 20 ms, p = 0.004; RA: 226 +/- 29 ms vs. 188 +/- 20 ms; p = 0.003). After AF exposure, the PVs demonstrated a significant conduction slowing whereas the atria did not (PVs: 125 +/- 33 ms vs. 159 +/- 37 ms, p < 0.001; LA: 129 +/- 26 ms vs. 130 +/- 24 ms, p = NS; RA: 192 +/- 36 ms vs. 196 +/- 32 ms, p = NS). Finally, AF was more frequently induced after the presence of AF, particularly by pacing in the PVs (14% vs. 49%, p = 0.001). CONCLUSIONS: New-onset, short-lasting AF creates electrical characteristics similar to those of patients with AF. However, these alterations are pronounced in the PVs compared with the atria, indicating that "AF begets AF in the PVs" (Electrophysiological Properties of the Pulmonary Veins; NCT00530608). PMID- 18510964 TI - The pulmonary veins speedy recoveries and early discharges. PMID- 18510965 TI - The right ventricular failure risk score a pre-operative tool for assessing the risk of right ventricular failure in left ventricular assist device candidates. AB - OBJECTIVES: This study sought to develop a model that estimates the post operative risk of right ventricular (RV) failure in left ventricular assist device (LVAD) candidates. BACKGROUND: Right ventricular failure after LVAD surgery is associated with increased morbidity and mortality, but identifying LVAD candidates at risk for RV failure remains difficult. METHODS: A prospectively collected LVAD database was evaluated for pre-operative clinical, laboratory, echocardiographic, and hemodynamic predictors of RV failure. Right ventricular failure was defined as the need for post-operative intravenous inotrope support for >14 days, inhaled nitric oxide for > or =48 h, right-sided circulatory support, or hospital discharge on an inotrope. An RV failure risk score (RVFRS) was created from multivariable logistic regression model coefficients, and a receiver-operating characteristic curve of the score was generated. RESULTS: Of 197 LVADs implanted, 68 (35%) were complicated by post operative RV failure. A vasopressor requirement (4 points), aspartate aminotransferase > or =80 IU/l (2 points), bilirubin > or =2.0 mg/dl (2.5 points), and creatinine > or =2.3 mg/dl (3 points) were independent predictors of RV failure. The odds ratio for RV failure for patients with an RVFRS < or =3.0, 4.0 to 5.0, and > or =5.5 were 0.49 (95% confidence interval [CI] 0.37 to 0.64), 2.8 (95% CI 1.4 to 5.9), and 7.6 (95% CI 3.4 to 17.1), respectively, and 180-day survivals were 90 +/- 3%, 80 +/- 8%, and 66 +/- 9%, respectively (log rank for linear trend p = 0.0045). The area under the receiver-operating characteristic curve for the RVFRS (0.73 +/- 0.04) was superior to that of other commonly used predictors of RV failure (all p < 0.05). CONCLUSIONS: The RVFRS, composed of routinely collected, noninvasive pre-operative clinical data, effectively stratifies the risk of RV failure and death after LVAD implantation. PMID- 18510966 TI - Plasma concentration of SCUBE1, a novel platelet protein, is elevated in patients with acute coronary syndrome and ischemic stroke. AB - OBJECTIVES: This study investigates the potential application of plasma SCUBE1 [signal peptide-CUB (complement C1r/C1s, Uegf, and Bmp1)-EGF (epidermal growth factor)-like domain-containing protein 1] as a biomarker of platelet activation in acute coronary syndrome (ACS) and acute ischemic stroke (AIS). BACKGROUND: Platelet activation plays a crucial role in ACS and AIS. Platelet stimulation is associated with increased plasma concentration of SCUBE1, a novel platelet endothelial secreted protein identified in our previous study. METHODS: Plasma concentrations of SCUBE1 from 40 ACS and 40 AIS patients were measured by enzyme linked immunoadsorbent assay and compared with the levels of 40 healthy control subjects and 83 chronic coronary artery disease (CAD) patients. Two-dimensional electrophoresis followed by Western blotting was used to characterize SCUBE1 protein in patients' plasma. RESULTS: Plasma SCUBE1 concentration was virtually undetectable in healthy control subjects and CAD patients, but was significantly higher in ACS and AIS patients (median = 205 and 95.1 ng/ml, respectively, p < 0.01). The increase in plasma SCUBE1 was detectable in the plasma as early as 6 h after the onset of symptoms and remained detectable up to 84 h. Plasma SCUBE1 concentration is an independent predictor of stroke severity based on National Institutes of Health Stroke Scale (beta = 3.18, p < 0.001). Furthermore, smaller SCUBE1 fragments were detected in ACS patients' plasma, suggesting that plasma SCUBE1 might subject to a proteolytic regulation under pathological conditions. CONCLUSIONS: Plasma SCUBE1 concentration is significantly elevated in ACS and AIS but not CAD patients. Plasma SCUBE1 is a potential biomarker of platelet activation in acute thrombotic disease. PMID- 18510967 TI - Will SCUBE1 solve the ischemia marker deficit? PMID- 18510968 TI - Defective intercellular adhesion complex in myocardium predisposes to infarct rupture in humans. AB - OBJECTIVES: Our goal was to evaluate intercellular adhesion complex proteins in myocardium in human infarct rupture. BACKGROUND: Infarct rupture, a fatal complication of myocardial infarction (MI), has been attributed to a defective cell adhesion complex in a transgenic mouse model. METHODS: Heart samples were collected from autopsies from infarct rupture and control (nonrupture) MI patients. Both infarcted and remote areas were included. Cell adhesion proteins including alphaE-catenin, beta-catenin, gamma-catenin, and N-cadherin were characterized by immunohistochemistry and immunoblotting. Genetic analysis was undertaken to evaluate mutations and polymorphisms in the alphaE-catenin gene. In addition, infarct rupture was studied in transgenic mice heterozygous for alphaE catenin C-terminal deficiency, mimicking the situation in human infarct rupture patients. RESULTS: No alphaE-catenin was detected in 70% of remote samples of infarct rupture hearts compared with 20% in control MI by immunohistochemistry. The immunoblot analysis confirmed a significant reduction in remote areas, and complete absence of alphaE-catenin in infarct areas from infarct rupture patients. No mutation or polymorphism of the alphaE-catenin gene was discovered. Other cell adhesion proteins were not significantly affected in remote areas of infarct rupture hearts. Three-fourths of the heterozygous alphaE-catenin C terminal truncated mice died of infarct rupture, compared with one-fourth of the wild-type littermates. CONCLUSIONS: The data show a reduced expression and defective localization of alphaE-catenin in the intercalated disc region in patients dying of infarct rupture. The mechanism of lower expression of alphaE catenin remains to be elucidated. PMID- 18510969 TI - President's page: The continuing importance of international engagement. PMID- 18510970 TI - Association of the 719Arg variant of KIF6 with both increased risk of coronary events and with greater response to statin therapy. PMID- 18510972 TI - Bacillus thuringiensis pore-forming toxins trigger massive shedding of GPI anchored aminopeptidase N from gypsy moth midgut epithelial cells. AB - The insecticidal Cry proteins produced by Bacillus thuringiensis strains are pore forming toxins (PFTs) that bind to the midgut brush border membrane and cause extensive damage to the midgut epithelial cells of susceptible insect larvae. Force-feeding B. thuringiensis PFTs to Lymantria dispar larvae elicited rapid and massive shedding of a glycosylphosphatidylinositol (GPI)-anchored aminopeptidase N (APN) from midgut epithelial cells into the luminal fluid, and depletion of the membrane-anchored enzyme on the midgut epithelial cells. The amount of APN released into the luminal fluid of intoxicated larvae was dose- and time dependent, and directly related to insecticidal potency of the PFTs. The induction of toxin-induced shedding of APN was inhibited by cyclic AMP and MAPK kinase (MEK) inhibitors PD98059 and U0126, indicating that signal transduction in the MEK/ERK pathway is involved in the regulation of the shedding process. APN released from epithelial cells appears to be generated by the action of a phosphatidylinositol-specific phospholipase C (PI-PLC) cleavage of the GPI anchor based upon detection of a cross-reacting determinant (CRD) on the protein shed into the luminal fluid. Alkaline phosphatase was also released from the gut epithelial cells, supporting the conclusion that other GPI-anchored proteins are released as a consequence of the activation PI-PLC. These observations are the basis of a novel and highly sensitive tool for evaluating the insecticidal activity of new Cry proteins obtained though discovery or protein engineering. PMID- 18510973 TI - The cuticular fatty acids of Calliphora vicina, Dendrolimus pini and Galleria mellonella larvae and their role in resistance to fungal infection. AB - Epicuticular lipids in many terrestrial arthropods consist of vast numbers of polar and non-polar aliphatic compounds, which are mainly responsible for the water balance in these animals but can also affect conidia germination of entomopathogenic fungi. In this work the qualitative and quantitative profiles of cuticular fatty acids from three insect species differing in their susceptibility to fungal infection were studied. In an innovative approach, laser light scattering detection was coupled with HPLC in order to identify the non chromophoric chemicals usually present in cuticular extracts. The acids identified contained from 5 to 20 carbon atoms in the alkyl chain and included unsaturated entities such as C(16:1), C(18:1), C(18:2), C(18:3) and C(20:1). There was a marked dominance of acids containing 16-18 carbon atoms. The relative contents of fatty acids in the extracted waxes varied from trace amounts to 44%. Cuticular fatty acids profile of Calliphora vicina (species resistant to fungal infection) significantly differs from profiles of Dendrolimus pini and Galleria mellonella (both species highly susceptible to fungal infection). The major difference is the presence of C(14:0), C(16:1) and C(20:0) in the cuticle of C. vicina. These three fatty acids are absent in the cuticle of D. pini while G. mellonella cuticle contains their traces. The concentrations of four fatty acids dominating in the G. mellonella larval cuticle (C(16:0), C(18:0), C(18:1) and C(18:2)) were found to fluctuate during the final larval instar and correlate with fluctuations in the susceptibility of larvae to fungal infection. The possible role of cuticular fatty acids in preventing fungal infection is discussed. PMID- 18510974 TI - Subsite substrate specificity of midgut insect chymotrypsins. AB - Insect chymotrypsins are distinctively sensitive to plant protein inhibitors, suggesting that they differ in subsite architecture and hence in substrate specificities. Purified digestive chymotrypsins from insects of three different orders were assayed with internally quenched fluorescent oligopeptides with three different amino acids at P1 (Tyr, Phe, and Leu) and 13 amino acid replacements in positions P1', P2, and P3. The binding energy (DeltaG(s), calculated from K(m) values) and the activation energy (DeltaG(T)++, determined from k(cat)/K(m) values) were calculated. The hydrophobicities of each subsite were calculated from the efficiency of hydrolysis of the different amino acid replacements at that subsite. The results showed that except for S1, the other subsites (S2, S3, and S1') vary among chymotrypsins. This result contrasts with insect trypsin data that revealed a trend along evolution, putatively associated with resistance to plant inhibitors. In spite of those differences, the data suggested that in lepidopteran chymotrypsins S2 and S1' bind the substrate ground state, whereas only S1' binds the transition state, supporting aspects of the present accepted mechanism of catalysis. PMID- 18510975 TI - Over-expression of cytochrome P450 CYP6CM1 is associated with high resistance to imidacloprid in the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae). AB - The two most damaging biotypes of Bemisia tabaci, B and Q, have both evolved strong resistance to the neonicotinoid insecticide imidacloprid. The major mechanism in all samples investigated so far appeared to be enhanced detoxification by cytochrome P450s monooxygenases (P450s). In this study, a polymerase chain reaction (PCR) technology using degenerate primers based on conserved P450 helix I and heme-binding regions was employed to identify P450 cDNA sequences in B. tabaci that might be involved in imidacloprid resistance. Eleven distinct P450 cDNA sequences were isolated and classified as members of the CYP4 or CYP6 families. The mRNA expression levels of all 11 genes were compared by real-time quantitative RT-PCR across nine B and Q field-derived strains of B. tabaci showing strong resistance, moderate resistance or susceptibility to imidacloprid. We found that constitutive over-expression (up to approximately 17-fold) of a single P450 gene, CYP6CM1, was tightly related to imidacloprid resistance in both the B and Q biotypes. Next, we identified three single-nucleotide polymorphic (SNP) markers in the intron region of CYP6CM1 that discriminate between the resistant and susceptible Q-biotype CYP6CM1 alleles (r-Q and s-Q, respectively), and used a heterogeneous strain to test for association between r-Q and resistance. While survivors of a low imidacloprid dose carried both the r-Q and s-Q alleles, approximately 95% of the survivors of a high imidacloprid dose carried only the r-Q allele. Together with previous evidence, the results reported here identify enhanced activity of P450s as the major mechanism of imidacloprid resistance in B. tabaci, and the CYP6CM1 gene as a leading target for DNA-based screening for resistance to imidacloprid and possibly other neonicotinoids in field populations. PMID- 18510976 TI - Metabolism of myristicin by Depressaria pastinacella CYP6AB3v2 and inhibition by its metabolite. AB - Although methylenedioxyphenyl (MDP) compounds, such as myristicin, are useful in the management of insecticide-resistant insects, the molecular mechanisms for their action in mammals and insects have not been elucidated. In this study, GC MS analyses of methanol extracts of foliage of wild parsnip (Pastinaca sativa) have identified myristicin as a substrate for CYP6AB3v2, an imperatorin metabolizing cytochrome P450 monooxygenase from Depressaria pastinacella (parsnip webworm). In contrast with its strong inhibitory effects on many mammalian P450s, myristicin is effectively metabolized by CYP6AB3v2 (V(max) and K(m) of 97.9 pmol/min/pmol P450 and 17.9 microM, respectively) at a rate exceeding that recorded previously for imperatorin, the only other known substrate for this highly specialized enzyme. The myristicin metabolite of CYP6AB3v2 is 1-(3',4' methylenedioxy-5'-methoxyphenyl)-2,3-epoxypropane. Molecular dockings have indicated that, unlike other epoxide metabolites of furanocoumarins, this epoxide metabolite is likely to remain in the CYP6AB3v2 catalytic site due to its low binding energy (-31.0 kcal/mol). Inhibition assays indicate that myristicin acts as a mixed inhibitor of this insect P450 and suggest that the epoxide metabolite may be an intermediate involved in the formation of P450-methylenedioxyphenyl complexes. PMID- 18510977 TI - Gene silencing in phlebotomine sand flies: Xanthine dehydrogenase knock down by dsRNA microinjections. AB - Lutzomyia longipalpis are vectors of medically important visceral leishmaniasis in South America. Blood-fed adult females digest large amounts of protein, and xanthine dehydrogenase is thought to be a key enzyme involved in protein catabolism through the production of urate. Large amounts of heme are also released during digestion with potentially damaging consequences, as heme can generate oxygen radicals that damage lipids, proteins and nucleic acids. However, urate is an antioxidant that may prevent such oxidative damage produced by heme. We investigated xanthine dehydrogenase by developing the RNAi technique for sand flies and used this technique to knock down the Lu. longipalpis xanthine dehydrogenase gene to evaluate its role in survival of adult females after blood feeding. The gene sequence of Lu. longipalpis xanthine dehydrogenase is described together with expression in different life cycle stages and RNAi knock down. Semi quantitative RT-PCR of xanthine dehydrogenase expression showed a significant increase in expression after bloodmeal ingestion. Microinjection of dsRNA via the thorax of 1-day-old adult female sand flies resulted in approximately 40% reduction of xanthine dehydrogenase gene expression in comparison to flies injected with a control dsRNA. A significant reduction of urate in the whole body and excretions of Lu. longipalpis was observed after dsRNA xanthine dehydrogenase microinjection and feeding 96h later on rabbit blood. Sand flies injected with XDH dsRNA also exhibit significantly reduced life span in comparison with the mock-injected group when fed on sucrose or when rabbit blood fed, showing that urate could be indeed an important free radical scavenger in Lu. longipalpis. The demonstration of xanthine dehydrogenase knock down by dsRNA microinjection, low mortality of microinjected insects and the successful bloodfeeding of injected insects demonstrated the utility of RNAi as a tool for functional analysis of genes in phlebotomine sand flies. PMID- 18510978 TI - Extensive gene amplification and concerted evolution within the CPR family of cuticular proteins in mosquitoes. AB - Annotation of the Anopheles gambiae genome has revealed a large increase in the number of genes encoding cuticular proteins with the Rebers and Riddiford Consensus (the CPR gene family) relative to Drosophila melanogaster. This increase reflects an expansion of the RR-2 group of CPR genes, particularly the amplification of sets of highly similar paralogs. Patterns of nucleotide variation indicate that extensive concerted evolution is occurring within these clusters. The pattern of concerted evolution is complex, however, as sequence similarity within clusters is uncorrelated with gene order and orientation, and no comparable clusters occur within similarly compact arrays of the RR-1 group in mosquitoes or in either group in D. melanogaster. The dearth of pseudogenes suggests that sequence clusters are maintained by selection for high gene-copy number, perhaps due to selection for high expression rates. This hypothesis is consistent with the apparently parallel evolution of compact gene architectures within sequence clusters relative to single-copy genes. We show that RR-2 proteins from sequence-cluster genes have complex repeats and extreme amino-acid compositions relative to single-copy CPR proteins in An. gambiae, and that the amino-acid composition of the N-terminal and C-terminal sequence flanking the chitin-binding consensus region evolves in a correlated fashion. PMID- 18510979 TI - Pyrosequence analysis of expressed sequence tags for Manduca sexta hemolymph proteins involved in immune responses. AB - The tobacco hornworm Manduca sexta is widely used as a model organism to investigate the biochemical basis of insect physiological processes but little transcriptome information is available. To get a broad view of the larval hemolymph proteins, particularly those related to immunity, we synthesized and sequenced cDNA fragments from a mixture of eight total RNA samples: fat body and hemocytes from larvae injected with killed bacteria, fat body, hemocytes, integument and trachea from naive larvae, and fat body and hemocytes from wandering larvae. Using massively parallel pyrosequencing, we obtained 95,458 M. sexta expressed sequence tags (ESTs) at an average size of 185bp per read. A majority of the sequences (69,429 reads) could be assembled into 7231 contigs with an average size of 300bp, 1178 of which had significant similarity with Drosophila genes from various functional groups. Only approximately 8% (606) of the contigs matched known M. sexta cDNA sequences, representing 186 of the 375 unique NCBI entries. The remaining 6625 contigs represented newly discovered cDNA segments from this well studied biochemical model insect. A search of the 7231 contigs using Tribolium castaneum, Drosophila melanogaster, and Bombyx mori immunity-related sequences revealed 424 cDNA contigs with significant similarity (E-value <1 x 10(-5)). These included 218 previously unknown M. sexta sequences coding for putative defense molecules such as pattern recognition receptors, serine proteinases, serpins, Spatzle, Toll-like receptors, intracellular signaling molecules, and antimicrobial peptides. PMID- 18510980 TI - Preserving public confidence. PMID- 18510981 TI - How not to treat cancer. PMID- 18510982 TI - Stellate-ganglion block: a new treatment for hot flushes? PMID- 18510983 TI - A smart move against cancer for vaccinia virus. PMID- 18510984 TI - Galectin-3 for indeterminate thyroid cytology. PMID- 18510985 TI - Periodontal diseases and cancer. PMID- 18510986 TI - Treating Burkitt's lymphoma in Malawi, Cameroon, and Ghana. PMID- 18510987 TI - Vulvovaginal reconstruction for neoplastic disease. AB - Current treatment of neoplastic disease that involves the external female genitalia aims to achieve local disease control, but not to restore form and function of these organs. Despite a growing trend to reduce the extent of surgical resection, impaired quality of life after surgery due to severe sexual dysfunction and disturbed body image is common. We postulate that the integration of surgical techniques for vulvar and vaginal reconstruction into primary treatment would improve aesthetic and functional results and therefore quality of life. We systematically searched the literature for surgical procedures designed and validated for vulvovaginal reconstruction. Various skin flaps, both with random vascularisation and those based on vascular territories (ie, axial pattern, fasciocutaneous, musculocutaneous, and bowel flaps), can restore important parts of vulvovaginal form and function with acceptable morbidity at the donor and recipient sites. Appropriate vulvovaginal reconstruction cannot be achieved by doing a few standardised procedures; rather, it necessitates specialists who are familiar with general principles of reconstructive surgery to master many techniques to select the optimum procedure for the individual patient. Vulvovaginal reconstructive surgery has limitations, particularly achievement of functional restoration in irradiated tissue. Physicians who treat women with neoplastic disease of the external genitalia should be aware of the current state of vulvovaginal reconstructive surgery. Prospective controlled clinical trials are warranted to assess the effect of vulvovaginal reconstruction on morbidity and quality of life after treatment. PMID- 18510988 TI - Role of the cerebellum in the neurocognitive sequelae of treatment of tumours of the posterior fossa: an update. AB - The lengthened survival of patients with tumours of the posterior fossa has brought awareness of the neurocognitive deficits present in this patient population. In the past, these deficits were thought to be caused by radiotherapy damaging supratentorial structures known to be responsible for cognitive processing. This notion led to the development of new treatment protocols to restrict damage to supratentorial regions by decreasing the radiation dose and the irradiated volume. However, these treatment protocols have only resulted in marginal improvements, sometimes at the expense of long-term survival. Moreover, the current published work reports that non-irradiated patients with tumours of the posterior fossa exhibit similar cognitive impairments to irradiated patients. The growth and treatment of tumours of the posterior fossa also damage infratentorial structures, including the cerebellum. Findings from anatomical, clinical, and neuroimaging studies support a role for the cerebellum in cognitive functions similar to those impaired in patients with a tumour of the posterior fossa. Despite these findings, research focused on the treatment of these patients and on decreasing their cognitive impairments either ignores that the cerebellum has been implicated in non-motor functions or argues against the possibility that damage to the cerebellum might result in cognitive sequelae. Future studies need to address the possibility that the cognitive impairments of patients with tumours of the posterior fossa might be determined by a combination of factors, including damage to the cerebellum. Recognition of the important cognitive contributions of the cerebellum might lead to improved cognitive outcome and quality of life for this patient population. PMID- 18510989 TI - Management of venous thromboembolism in patients with advanced cancer: a systematic review and meta-analysis. AB - Venous thromboembolism is common in patients with cancer. However, no management guidelines exist for venous thromboembolism specific to patients with advanced progressive cancer. To help develop recommendations for practice, we have done a comprehensive review of anticoagulation treatment in patients with cancer, with particular focus on studies that included patients with advanced disease. Data from 19 publications, including randomised, prospective, and retrospective studies suggest that: long-term full-dose low-molecular-weight heparin (LMWH) is more effective than warfarin in the secondary prophylaxis of venous thromboembolism in patients with cancer of any stage, performance status, or prognosis; warfarin should not be used in patients with advancing progressive disease; and in patients at high risk of bleeding, full-dose LMWH for 7 days followed by a long-term decreased fixed dose long term can be considered. The optimum treatment duration is unclear, but because the prothrombotic tendency will persist in patients with advanced cancer, indefinite treatment is generally recommended. For patients with contraindications to anticoagulation, inferior vena-caval filters can be considered, but their use needs careful patient selection. Ultimately, the decision to initiate, continue, and stop anticoagulation will need to be made on an individual basis, guided by the available evidence, the patient's circumstances, and their informed preferences. PMID- 18510990 TI - Incidence, aetiology, and outcomes of cancer in Indigenous peoples in Australia. AB - An assessment of recent data on cancer in Indigenous Australians (Aborigines and Torres Strait Islanders) shows that, although they are less likely to have some types of cancer than other Australians, Indigenous people are significantly more likely to have cancers that have a poor prognosis, but are largely preventable, such as lung and liver cancer. Indigenous people with cancer are diagnosed at a later stage, are less likely to receive adequate treatment, and are more likely to die from their cancers than other Australians. Inadequate identification of Indigenous people in cancer registers precludes reporting for some parts of Australia, but sufficient information is available to identify priorities and inform appropriate remedial action. Health-risk factors, especially smoking, and inadequate health-system performance largely explain the patterns of cancer incidence and mortality in areas with adequate data. Effective tobacco control programmes, improvements across a range of health services, and meaningful Indigenous engagement are all needed to decrease the burden of cancer in Indigenous Australians. PMID- 18510991 TI - Carcinoma of unknown primary with a colon-cancer profile-changing paradigm and emerging definitions. AB - Carcinoma of unknown primary (CUP), which accounts for about 3-5% of all new cancers, is a challenging heterogeneous entity with an unmet research need. Traditionally, CUP has been managed with broad-spectrum chemotherapy, but with the increasing availability of sophisticated diagnostic techniques and the emergence of new treatments that have been shown to be effective in specific cancers the one-treatment-fits-all approach to CUP might eventually no longer be valid. CUP in association with a colon-cancer profile (CCP-CUP) is an example of an emerging, specific CUP subset that seems to benefit from a tailored approach. CCP-CUP is identified by CK20 and CDX2-positive and CK7-negative immunohistochemistry and a clinical course consistent with that of patients known to have metastatic colon cancer. Our findings suggest that patients with CCP-CUP derive substantial benefit from the use of specific treatments developed for colon cancer and larger clinical trials are warranted to more definitely test this finding. In the era of molecular profiling, we expect that additional work with CCP-CUP and other CUP subsets will provide attractive tailored treatment alternatives, with efficacies that exceed the current one-treatment-fits-all approach. PMID- 18510992 TI - Part I: Milestones in personalised medicine--imatinib. PMID- 18510993 TI - Effect of deleterious nsSNP on the HER2 receptor based on stability and binding affinity with herceptin: a computational approach. AB - In this study, we identified the most deleterious non-synonymous SNP of ERBB2 (HER2) receptors by its stability and investigated its binding affinity with herceptin. Out of 135 SNPs, 10 are nsSNPs in the coding region, in which one of the nsSNP (SNPid rs4252633) is commonly found to be damaged by I-Mutant 2.0, SIFT and PolyPhen servers. With this effort, we modelled the mutant HER2 protein based on this deleterious nsSNP (rs4252633). The modeled mutant showed less stability than native HER 2 protein, based on both total energy of the mutant and stabilizing residues in the mutant protein. This is due to a deviation between the mutant and the native HER2, having an RMSD of about 2.81 A. Furthermore, we compared the binding efficiency of herceptin with native and mutant HER2 receptors. We found that herceptin has a high binding affinity with mutant HER2 receptor, with a binding energy of -24.40 kcal/mol, as compared to the native type, which has a binding energy of -15.26 kcal/mol due to six-hydrogen bonding and two salt bridges exist between herceptin and the mutant type, whereas the native type establishes four hydrogen bonds and two salt bridges with herceptin. This analysis portrays that mutant type has two additional hydrogen bonds with herceptin compared with the native type. Normal mode analysis also showed that the two amino acids, namely Asp596 and Glu598 of mutant HER2, forming additional hydrogen bonding with herceptin, had a slightly higher flexibility than the native type. Based on our investigations, we propose that SNPid rs4252633 could be the most deleterious nsSNP for HER2 receptor, and that herceptin could be the best drug for mutant compared to the native HER2 target. PMID- 18510994 TI - Osmoregulation and antioxidant metabolism in drought-stressed Helianthus annuus under triadimefon drenching. AB - A pot-culture experiment was conducted to estimate the ameliorating effect of triadimefon (TDM) on drought stress in sunflower (Helianthus annuus L.) plants. The plants were subjected to 3-, 6-, and 9-day-interval drought (DID) stress and drought stress with TDM @ 15 mg l(-1) and 15 mg l(-1) TDM alone from the 30th day after sowing (DAS). One-day-interval irrigation was kept as control. The plant samples were collected on and separated into root, stem and leaf for estimating the amino acid (AA), proline (PRO) and glycine betaine (GB) contents and the activities of antioxidant enzymes. Individual and combined drought stress and TDM treatments increased AA, PRO and GB contents, superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and polyphenol oxidase (PPO) activities when compared to control. From the results of this investigation, it can be concluded that the application of TDM caused a partial amelioration of the adverse effects of drought stress by its influence on quaternary ammonium compounds and antioxidant potentials in H. annuus plants. PMID- 18510995 TI - Cadmium-induced oxidative stress and DNA damage in kidney of pregnant female rats. AB - In the present study, we have investigated the influence of sub-acute treatment with cadmium (Cd) on some parameters indicative of oxidative stress and DNA damage in tissues of pregnant female rats. Pregnant female rats (n=6) were injected subcutaneously, daily with a dose of cadmium chloride of 3 mg/kg body weight (b.w.) from day 6 to day 19 of pregnancy, and they were allowed to deliver normally. MDA level and GPx, CAT and SOD activities were used as markers of oxidative stress in liver and kidney. The 8-oxo-dG level was measured by the HPLC EC system. Cd treatment increased MDA (+116%, p<0.01) in kidney. Moreover, Cd treatment also decreased CuZn-SOD (-11%, p<0.05) and GSH level (-52%, p<0.05) in kidney. Treated rats displayed an increase of the liver metallothionein (MT) level. Induction of MT in liver was probably implicated in the detoxification of Cd. The high level of Cd (3 mg/kg) used in the present study is partially neutralized by MT in liver, whereas the free fraction could be implicated in the oxidative stress and DNA oxidation observed in kidney. Cd treatment failed to alter 8-oxodGuo, indicating the absence of DNA oxidation in liver; by contrast, the same treatment increased the 8-oxodGuo level (+51%, p<0.05) in the kidney of pregnant female rats, indicating an oxidative stress associated with DNA damage only in kidney. PMID- 18510996 TI - Higher plant antioxidants and redox signaling under environmental stresses. AB - Main antioxidants in higher plants include glutathione, ascorbate, tocopherol, proline, betaine, and others, which are also information-rich redox buffers and important redox signaling components that interact with biomembrane-related compartments. As an evolutionary consequence of aerobic life for higher plants, reactive oxygen species (ROS) are formed by partial reduction of molecular oxygen. The above enzymatic and non-enzymatic antioxidants in higher plants can protect their cells from oxidative damage by scavenging ROS. In addition to crucial roles in defense system and as enzyme cofactors, antioxidants influence higher plant growth and development by modifying processes from mitosis and cell elongation to senescence and death. Most importantly, they provide essential information on cellular redox state, and regulate gene expression associated with biotic and abiotic stress responses to optimize defense and survival. An overview of the literature is presented in terms of main antioxidants and redox signaling in plant cells. Special attention is given to ROS and ROS-antioxidant interaction as a metabolic interface for different types of signals derived from metabolism and from the changing environment, which regulates the appropriate induction of acclimation processes or, execution of cell death programs, which are the two essential directions for higher plants. PMID- 18510997 TI - Combined effects of long-term salinity and soil drying on growth, water relations, nutrient status and proline accumulation of Sesuvium portulacastrum. AB - The interaction between soil drying and salinity was studied in the perennial halophyte, Sesuvium portulacastrum. Rooted cuttings were individually cultivated for three months in silty-sandy soil under two irrigation modes: 100 and 25% of field capacity (FC). The amount of the evapotranspirated water was replaced by a nutrient solution containing either 0 or 100 mM NaCl. Whole-plant growth, leaf water content, leaf water potential (Psi(w)), and Na+, K+, and proline concentrations in the tissues were measured. When individually applied, both drought and salinity significantly restricted whole-plant growth, with a more marked effect of the former stress. However, the effects of the two stresses were not additive on whole-plant biomass or on leaf expansion. Root growth was more sensitive to salt than to soil drying, the latter being even magnified by the adverse impact of salinity. Leaf water content was significantly reduced following exposure to water-deficit stress, but was less affected in salt-treated plants. When simultaneously submitted to water-deficit stress and salinity, plants displayed higher values of water and potassium use efficiencies, leaf proline and Na+ concentrations, associated with lower leaf water potential (-1.87 MPa), suggesting the ability of S. portulacastrum to use Na+ and proline for osmotic adjustment. PMID- 18510998 TI - Gardens in urbanizing rural areas reveal an unexpected floral diversity related to housing density. AB - The increasing urbanization of rural areas leads to a strong development of horticultural flora, which is the main source of alien and invasive plants. In order to assess the pool of cultivated species under different urbanization pressures, the diversity and distribution of horticultural flora were studied in 120 Mediterranean gardens belonging to three housing density types. The results showed a great richness and heterogeneity of this flora, and similarities in species composition between gardens of the same housing density types. Twenty four percent of the cultivated species are well adapted to the Mediterranean climate, and 21 species known to be invasive on the French territory have emanated from gardens. Inventorying areas adjoining gardens would be useful in identifying escaped garden plants and to assess the associated risks for biological diversity. The results also suggested a detailed analysis of the influence of social, economic and regional factors on planting practices, in order to identify the drivers of these original floral patterns. PMID- 18510999 TI - [Contribution to the study of land-use dynamics in the plains of Macta (Algeria) with the aid of remote sensing and GIS]. AB - The plains of Macta, where the Macta marshes are located, form a wetland with particular ecological characteristics in the Maghreb and are subject to distinct land use dynamics. The plant formations are exposed to multiple forms of degradation, primarily due to climate (period of dryness) and to increasing anthropogenic activities in the area. The new methods of breeding and cultivation contribute to ongoing degradation in this natural ecosystem, with a deterioration of the wetland's biological potentialities. With the aim of safeguarding Macta's marshes, a better knowledge of vegetation distribution patterns and dynamics, together with the interpretation of the remote sensing data and GIS, makes it possible to consider programmes of natural environment restoration and sustainable management. The results of land-use dynamics analysis show a decline in agricultural land use from 33,000 ha in 1958 to 21,000 ha in 2005. Halophytic vegetation currently occupies 47,000 ha, while in 1958 it occupied only 27,000 ha. Tamarix, Inula, Joncus and Scirpus species are experiencing a decrease in surface cover, while Salicornia, Suaeda, and Atriplex are extending their area. To cite this article: PMID- 18511000 TI - A glimpse of lignicolous marine fungi occurring in coastal water bodies of Tamil Nadu (India). AB - In the present investigation, a total of 51 marine fungi were obtained from wood samples collected from four locations of Tamil Nadu (Tuthukudi, Chennai, Kanyakumari and Pichavaram), India. Out of these 51, 28 were ascomycetes, one was basidiomycete and 22 were mitosporic fungi. Maximum fungal diversity was encountered from Tuthukudi, followed by Chennai, Kanyakumari, and the minimum from Pichavaram. Periconia prolifica was the only species common to all the four locations. PMID- 18511001 TI - [Biodiversity of the aquatic invertebrates of the eastern part of the Bay and the Estuary of the Seine River: the CISA database, two centuries of observations]. AB - An inventory of all aquatic invertebrates of the Estuary and the eastern part of the Bay of Seine (Normandy, France) was performed and integrated in a free-of charge georeferenced database named CISA (Catalogue of the Invertebrates in Seine Aval, e.g., the downstream part of the Seine from the Poses dam to the sea). One thousand four hundred eighty-five taxa of aquatic invertebrates, out of which 5% were never identified at the species level, were recorded. This inventory has been analyzed in comparison with observation efforts carried out in this zone since two centuries, by distinguishing the first inventories of the end of the 19th century, the publications of the regional naturalist societies, the scientific publications, and the grey literature, such as the scientific reports and the university works with limited diffusion. It appears that the regional naturalist societies have played an essential role for the knowledge of the biological diversity before World War II; since then, reports and university works mention approximately half of the new species for the region. Less than one quarter of these descriptions was thus the subject of a publication without restricted diffusion. Biological diversity was thus underestimated for a long time for the eastern part of the Bay and the Estuary of the Seine River. These data show that the distribution of invertebrate species in the estuary of the Seine follows a two-ecocline model. PMID- 18511002 TI - Forensic Epidemiology: a systematic approach to probabilistic determinations in disputed matters. AB - Forensic medicine testimony often relies upon terms of probability to enhance the strength of the testimony. Such terms must have a demonstrably reliable and accurate basis; otherwise their use is speculative, unjustified, and potentially harmful. Forensic Epidemiology is introduced as a framework from which probabilistic testimony can be assessed in settings in which it is either proffered or encountered. In this paper, common forensic uses of probability are reviewed, appropriate methods for presenting such testimony are proposed, and inappropriate uses of probability and epidemiologic concepts and data, as well as a logical fallacies commonly observed in forensic settings are presented. A previously unpublished logical fallacy, the "Prior Odds" Fallacy, is also introduced. PMID- 18511003 TI - Drug facilitated sexual assault--a review. AB - This review was undertaken to identify the evolutionary process in the current understanding of allegations of drug facilitated sexual assault (DFSA), so that those who work in this field may gain a better understanding of the complexities involved in such cases. Several definitions of DFSA are provided as well as a list of intoxicating substances which have so far been incriminated in this crime. Perception and alcohol use is addressed, whilst an examination of intoxication and victim outcomes reveals disturbing but important information which needs to be centrally placed within health education campaigns with a degree of urgency. The review identifies the effects of alcohol on sexual behaviour, drinking patterns and specific quantitative research indicating very high alcohol levels in some instances. In practical terms, suggestions are made following Operation Matisse, to address prevention, early detection and easier identification of DFSA cases so that victims' needs are prioritised and appropriately addressed. PMID- 18511004 TI - Injuries to neck structures in deaths due to constriction of neck, with a special reference to hanging. AB - This prospective study aimed at examining various injuries to the neck structures in deaths due to constriction of neck. Neck dissection technique, as advocated by Prinsloo and Gordon was undertaken to study the injuries to the thyro-hyoid complex, strap muscles, carotid vessels, etc. Of the 1746 medico-legal autopsies, conducted during the study period, 5% were deaths due asphyxia of which 82% were those of constriction of neck. The 21-30 years age group accounted for the maximum number of cases (57%). Male:female ratio was 2:1. Hanging (69%) outnumbered other asphyxial deaths--ligature and/or manual strangulation, smothering, etc. Injury to the sternocleido-mastoid muscle (54%) was the commonest injury to the neck structures. The hyoid bone was fractured in 21% cases, while the thyroid cartilage was fractured in 17% cases. Complete hanging was noted in 68% of cases while the hanging was atypical in 88%. Fixed knot was found to have been used in 71%. A single loop round the neck was observed in 80% of the cases and it was above the level of thyroid in 58% cases. Most cases of the fracture of the laryngo-hyoid complex were in the 41-60 year age group, 72% and the fracture was on the same side as the knot in 52% cases. Majority used soft daily wear articles of clothing like a sari (32%) or chunni (24%). Asphyxial deaths due to constriction of neck being common in all parts of the world, prospective studies in different setups to examine the profile of neck structure injuries are needed so as to differentiate the suicidal or homicidal nature of such deaths with a greater certainty. PMID- 18511005 TI - Changes in genital injury patterns over time in women after consensual intercourse. AB - To date, there are no studies in the literature addressing whether or not microscopic genital injuries change over time or change in appearance during the 72 h time period following intercourse. In this study, women (n=35) had two evidentiary type pelvic examinations to document injuries after consensual intercourse. At Time 1 (within 48 h of consensual intercourse) a: larger total surface area of injury (p=0.02); larger surface area of injury to the posterior fourchette (p=0.02); larger surface area of abrasions (p=0.04); and larger surface area of redness (p=0.04) were found compared to Time 2 (24 h after Time 1). Since this research is exploratory, larger studies are needed to explore the differences in genital injuries based on the time of examination and in women after non-consensual intercourse. PMID- 18511006 TI - Biochemical blood markers and sampling sites in forensic autopsy. AB - Forensic pathologists often hesitate to use biochemical blood markers due to the risk of large postmortem changes and deviations from healthy subjects. Biochemical analyses of postmortem blood, if possible, may help to evaluate pathological status and determining the cause of death in forensic diagnosis, for example, in sudden unexpected death without obvious cause, or young adults with no apparent cause of death or antemortem information. Even commercially available biochemical markers were re-evaluated in the blood samples of 164 forensic autopsy cases. Biochemical markers examined were HbA1c, fructosamine, blood nitrogen urea (BUN), creatinine, total protein, total bilirubin, gamma-glutamyl transpeptidase (gamma-GTP), triglyceride, total cholesterol, C-reactive protein (CRP) and pseudocholine esterase (pChE). We collected cardiac blood (left cardiac blood and right cardiac blood) and peripheral blood (femoral vein blood) to clarify the differences in measured values by sampling site. The measured values were analyzed in relation to postmortem interval, etiology of death and sampling sites. Of all eleven markers, HbA1c is the most useful and reliable because of its negligible postmortem changes and small deviation from healthy subjects. Total bilirubin, BUN, CRP and total cholesterol can be useful if we set appropriate limit ranges and pay attention to the interpretation. For the evaluation of changes due to postmortem intervals, none of the markers except for triglyceride showed significant changes up to three days postmortem. As for sampling sites, femoral vein blood is generally recommended considering postmortem changes, but left cardiac blood was suitable for creatinine, pChE, and total cholesterol. For clinical forensic diagnosis of biochemical blood markers, we must determine the "forensic abnormal value" after collecting more cases by known causes with more information about the population. PMID- 18511007 TI - Homeless deaths in Istanbul, Turkey. AB - We retrospectively analyzed the autopsy records of the Council of Forensic Medicine during the 5-year period between the years 2000 and 2004 to contribute to the efforts targeted at lowering death rate in the milieu of homelessness by documenting the current status of this group particularly in terms of mode of death. Two hundred and nine of the 229 cases (91.27%) were males and remaining 20 were females. The preponderance of male cases in our autopsy population was also detected in homeless population. Most of the cases were in the age group of 40-49 years (64 cases, 27.95%). One hundred and ninety two corpses (83.85%) were found outdoors and only 37 of the cases (16.15%) indoors. Natural events constituted the cause of death in 138 of the cases (60.26%) and in remainder 91 cases the cause of death was related to an unnatural event (39.74%). In approximately 1/3 of the cases of natural death cases, autopsy revealed the evidence of pulmonary tuberculosis but only in 32 of these cases the tuberculosis was the primary cause of death. The presence of alcohol was found to be significantly associated with the manner of death. Blood alcohol level over 50mg/100ml was determined only in 9.42% of natural death cases, whereas it was positive in 61.53% of unnatural death cases. We concluded that immediate precautions targeted at lowering death rate in this population must include health care for preventable natural diseases. PMID- 18511008 TI - Prevalence of pulmonary thromboemboli among referred cadavers having hospitalization records to Tehran Legal Medicine Center. AB - BACKGROUND: Pulmonary thromboemboli are one of the main causes of sudden death especially in hospitalized patients and appeared with different nonspecific manifestations. The aim of this study was to determine the prevalence of thromboemboli. MATERIALS AND METHODS: In this cross sectional study, pulmonary autopsies of 200 cadavers who were selected randomly from all cadavers with clinical suspicion of thromboemboli referred to Tehran University Tissue Archive in different months from January 2005 to 2006 and the prevalence of pulmonary embolism in these cases and its relation with demographic characteristics and sources of disease was assessed. Also, agreement degree of clinical and histopathological diagnosis of pulmonary embolism was calculated. RESULTS: The prevalence of pulmonary embolism was estimated at 13.5%. There were positive relationship between prevalence of pulmonary embolism and increased of age (P=0.001). Interpretation of results of macroscopic and histopathological studies for diagnosis of embolism showed moderate agreement (kappa=0.59) and interpretation of results of clinical diagnosis of disease before death and pathologic findings after death showed poor agreement (kappa=0.34). The most frequent detected location of emboli were end branches of pulmonary artery. CONCLUSION: Considering the apparent high prevalence of pulmonary embolism in our study, we recommend increased use of anti-deep vein thrombosis measures in all appropriate patients within the Tehran hospital population, according to evidence based guidelines. PMID- 18511009 TI - The value of subendocardial haemorrhages as an indicator of exsanguination and brain injury--a retrospective forensic autopsy study. AB - INTRODUCTION: Subendocardial hemorrhages (SE) have been associated with multiple causes of violent and non violent deaths in forensic autopsies such as fatal exsanguination, brain injury and intoxications. METHODS: The presented retrospective study investigates the overall incidence and various causes by an analysis of a total of 1331 forensic autopsies based on autopsy reports. RESULTS: The results show that head injury and significant blood loss alone or in combination are the main mechanisms of death associated with SE. The incidences of SE were 50% in exsanguination, 80% in combined exsanguination and head injury, and 31% in solitary head trauma. DISCUSSION: On the basis of the study results it can be concluded, that SE are important indicative signs for both fatal exsanguination and brain injury at medico-legal autopsies. PMID- 18511010 TI - Systematic evaluation of sensitivity and specificity of sibship determination by using 15 STR loci. AB - Paternity disputes and other forms of kinship testing are routinely resolved using short tandem repeat (STR) DNA loci. Sibship determination is encountered in instances where the DNA profiles of two individuals are compared to determine if they are siblings. If either parent is available for testing then the situation is simplified but if neither parent of the two individuals is available for DNA testing, a combined sibling indices (CSI) for the determination of sibship between two people can be determined. Support for kinship is also based upon the sharing of alleles, particularly when both alleles are shared at the same locus, termed two-allele-sharing-loci (TASL). We report on the combination of CSI and TASL to enhance the determination of sibship. The 15 STR loci that comprise the Identifiler loci were applied to three populations using pairs of full siblings or unrelated pairs. Based upon the data obtained, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) can be applied to determining whether two DNA profiles come from full or non-sibling pairs. This report highlights the problems inherent in this form of kinship testing and recommends a combination use of CSI and TASL for sibship determination. PMID- 18511011 TI - Assessing changes in pupillary size in Rifian smokers of kif (Cannabis sativa L.). AB - Although the measurement of eye pupil variations is a common method in the only few cannabis effect research, there are no studies on short term effects of kif (Moroccan traditional preparation of cannabis) on eye pupil. The aim of the present paper is to present results about effect of a smoked kif preparation (Cannabis sativa L.) on pupil diameter variations after 30 mn. Two examiners measured the pupil diameter variations before and after kif smoking in 34 eyes of 17 volunteer-consumers in a dark closed room. Pupil diameter was estimated by Colvard pupillometer. Results reveal a significantly increase in pupil size post kif. PMID- 18511012 TI - Automobile door entrapment--a different form of vehicle-related crush asphyxia. AB - Crush asphyxia involving motor vehicles usually occurs when a victim is trapped beneath a vehicle that slips from a jack while being worked on, or beneath a car that has rolled over during a crash. Two cases are reported where crush asphyxia resulted from quite different circumstances. Case 1: A 58-year-old woman was found dead trapped between her car door and frame. As she was alighting from the vehicle it had rolled forward trapping her between the semi-opened door and car frame when the door had wedged against a second parked vehicle. Case 2: A second 58-year-old woman was found dead, also trapped between her car door and frame. She had been leaning out of her car trying to pick up a newspaper when the car rolled forward. The open driver's door wedged against an adjacent pillar trapping her between the door and the frame. Both victims died from crush asphyxia. This form of automobile door entrapment represents a distinct subset of automobile related asphyxial deaths and illustrates a particular and unusual set of circumstances that may result in unexpected traumatic death. Getting, or leaning, out of a vehicle that does not have the handbrake engaged may result in wedging of the victim between the semi-opened door and car frame if the car rolls forward and the door impacts against a nearby unyielding object. Correlation of the physical dimensions of the door and frame with markings on the victim's body will assist in reconstructing the terminal events. PMID- 18511013 TI - A mystery: one wound, multiple bullets. AB - This case report provides an unusual presentation of a gunshot wound (GSW) and stresses the importance of gathering complete clinical, scene and historical information, if possible. Sufficient details regarding an injured patient's mechanism of injury (MOI) should be elicited by the treating physician when hemodynamic status of the patient allows. A careful physical exam is essential as are appropriate laboratory investigations and diagnostic imaging. We present a case report of a single GSW found on physical exam with multiple projectiles found on imaging studies. The history of present illness, scene findings and trial transcripts clarify the patient presentation. PMID- 18511014 TI - Suicides in India: a response to "Suicide in India--a 78 four year retrospective study" [J Forensic Leg Med 2007;14:185-9]. PMID- 18511015 TI - Mapping the surface (hydr)oxo-groups of titanium oxide and its interface with an aqueous solution: the state of the art and a new approach. AB - In this article the "titanium oxide/electrolyte solution" interface is studied by taking in advantage the recent developments in the field of Surface and Interface Chemistry relevant to this oxide. Ab-initio calculations were performed in the frame of the DFT theory for estimating the charge of the titanium and oxygen atoms exposed on the anatase (1 0 1), (1 0 0), (0 0 1), (1 0 3)(f) and rutile (1 1 0) crystal faces. These orientations have smaller surface energy with respect to other ones and thus it is more probable to be the real terminations of the anatase and rutile nanocrystallites in the titania polycrystalline powders. Potentiometric titrations for obtaining "fine structured" titration curves as well as microelectrophoresis and streaming potential measurements have been performed. On the basis of ab-initio calculations, and taking into account the relative contribution of each crystal face to the whole surface of the nanocrystals involved in the titania aggregates of a suspension, the three most probable surface ionization models have been derived. These models and the Music model are then tested in conjunction with the "Stern-Gouy-Chapman" and "Basic Stern" electrostatic models. The finally selected surface ionization model (model A) in combination with each one of the two electrostatic models describes very well the protonation/deprotonation behavior of titania. The description is also very good if this model is combined with the Three Plane (TP) model. The application of the "A/(TP)" model allowed mapping the surface (hydr)oxo-groups [TiO(H) and Ti(2)O(H)] of titania exposed in aqueous solutions. At pH>pzc almost all terminal oxygens [TiO] are non-protonated whereas even at low pH values the non-protonated terminal oxygens predominate. The acid-base behavior of the bridging oxygens [Ti(2)O] is different. Thus, even at pH=10 the greater portion of them is protonated. The application of the "A/TP" model in conjunction with potentiometric titrations, microelectrophoresis and streaming potential experiments allowed mapping the "titania/electrolyte solution" interface. It was found that the first (second) charged plane is located on the oxygen atoms of the first (second) water overlayer at a distance of 1.7 (3.4) A from the surface. The region between the surface and the second plane is the compact layer. The region between the second plane and the shear plane is the stagnant diffuse part of the interface, with an ionic strength dependent width, ranging from 20 (0.01 M) up to 4 A (0.3 M). The region between the shear plane and the bulk solution is the mobile diffuse part, with an ionic strength dependent width, ranging from 10 (0.01 M) up to 2 A (0.3 M). At I>0.017 M the mean concentration of the counter ions is higher in the stagnant than in the mobile part of the diffuse layer. For a given I, removal of pH from pzc brings about an increase of the mean concentration in the interfacial region and a displacement of the counter ions from the mobile to the stagnant part of the diffuse layer. The mean concentration of the counter ions in the compact layer is generally lower than the corresponding ones in the stagnant and mobile diffuse layers. The mobility of the counter ions in the stagnant layer decreases as pH draws away from pzc or ionic strength increases. PMID- 18511016 TI - Metabolic syndrome in postmenopausal women: the influence of oral or transdermal estradiol on inflammation and coagulation markers. AB - OBJECTIVE: The objective of the study was to determine whether the route of administration of estrogen therapy in women with metabolic syndrome (MBS) influences inflammation and coagulation parameters. STUDY DESIGN: Fifty symptomatic postmenopausal women with MBS were randomized to receive 1 mg oral estradiol (oE(2)) or 0.05 mg transdermal E(2) (tE(2)) for 3 months. Measurements were compared with those of 20 healthy premenopausal women and 74 normal postmenopausal women. RESULTS: Compared with both control groups, women with MBS had significantly higher levels of certain inflammation and coagulation markers, which cannot be accounted for based on weight alone. After oE(2), antithrombin III decreased from 104% to 96% (P < .01), the metalloproteinase-9/ tissue inhibitor of metalloproteinase-1 ratio increased (P < .02), and E-selectin decreased from 60 +/- 4.4 to 55 +/- 4.6 ng/mL (P < .05). With tE(2), there were no major changes noted. CONCLUSION: Postmenopausal women with MBS have higher levels of certain coagulation and inflammation markers and different responses to oral compared with transdermal estradiol. PMID- 18511017 TI - The role of radiation in improving survival for early-stage carcinosarcoma and leiomyosarcoma. AB - OBJECTIVE: We examined the effect of radiation on survival for early-stage uterine carcinosarcomas and leiomyosarcomas. STUDY DESIGN: The surveillance, epidemiology, and end results database was used to identify patients with stage I/II carcinosarcomas and leiomyosarcomas. Logistic regression and Cox models were developed to determine radiation use and survival. RESULTS: Among 1819 women with carcinosarcomas and 1088 women with leiomyosarcomas, radiation was administered to 667 of the patients (37%) with carcinosarcomas and to 235 of the patients (22%) with leiomyosarcomas. In a multivariate model, adjuvant radiation reduced the risk of death by 21% in women with carcinosarcomas (hazard ratio, 0.79; 95% CI, 0.7-0.9). Radiation reduced mortality rates in patients with carcinosarcomas who had not undergone node dissection but had only a marginal effect on survival in node-negative women. Adjuvant radiation had no effect on survival for early stage leiomyosarcomas (hazard ratio, 1.1; 95% CI, 0.9-1.4). CONCLUSION: Adjuvant radiotherapy improves survival for select patients with early-stage carcinosarcomas but is of limited value for leiomyosarcomas. PMID- 18511018 TI - Maternal morbidity following a trial of labor after cesarean section vs elective repeat cesarean delivery: a systematic review with metaanalysis. AB - This study reviewed maternal morbidity following trial of labor (TOL) after cesarean section, compared with elective repeat cesarean delivery (ERCS). Articles were pooled to compare women planning vaginal birth after cesarean (VBAC) with those undergoing ERCS with regard to maternal morbidity (MM), uterine rupture/dehiscence (UR/D), blood transfusion (BT), and hysterectomy. The former group was subdivided into successful VBAC (S-VBAC) and failed TOL (F-TOL). VBAC was successful in 17,905 of 24,349 patients (73%). MM, BT, and hysterectomy were similar in women planning VBAC or ERCS, whereas UR/D was different (1.3%; 0,4%). MM, UR/D, BT and hysterectomy were more common after F-TOL (17%, 4.4%, 3%; 0.5%) than after S-VBAC (3.1%, 0.2%, 1.1%; 0.1%) or ERCS (4.3%, 0.4%, 1%; 0.3%). Outcomes were more favorable in S-VBAC than ERCS. These findings show that a higher risk of UR/D in women planning VBAC than ERCS is counterbalanced by reduction of MM, UR/D. and hysterectomy when VBAC is successful. PMID- 18511019 TI - Chronic electroconvulsive stimulation but not chronic restraint stress modulates mRNA expression of voltage-dependent potassium channels Kv7.2 and Kv11.1 in the rat piriform cortex. AB - The mechanisms by which stress and electroconvulsive therapy exert opposite effects on the course of major depression are not known. Potential candidates might include the voltage-dependent potassium channels. Potassium channels play an important role in maintaining the resting membrane potential and controlling neuronal excitability. To explore this hypothesis, we examined the effects of one or several electroconvulsive stimulations and chronic restraint stress (6 h/day for 21 days) on the expression of voltage-dependent potassium channel Kv7.2, Kv11.1, and Kv11.3 mRNA in the rat brain using in situ hybridization. Repeated, but not acute, electroconvulsive stimulation increased Kv7.2 and Kv11.1 mRNA levels in the piriform cortex. In contrast, restraint stress had no significant effect on mRNA expression of Kv7.2, Kv11.1, or Kv11.3 in any of the brain regions examined. Thus, it appears that the investigated voltage-dependent potassium channels are not modulated by restraint stress at the level of mRNA expression. However, our findings suggest that repeated electroconvulsive stimulation alter Kv7.2 and Kv11.1 function in the piriform cortex, a finding with potential relevance for the chain of neurobiological events underlying the clinical effects of ECT. PMID- 18511020 TI - Pyramidal neurons in the septal and temporal CA1 field of the human and hedgehog tenrec hippocampus. AB - The present study examines comparatively the cellular density of disector counted/Nissl-stained CA1 pyramidal neurons and the morphometric characteristics (dendritic number/length, spine number/density and Sholl-counted dendritic branch points/20 microm) of the basal and apical dendritic systems of Golgi-impregnated CA1 neurons, in the septal and temporal hippocampus of the human and hedgehog tenrec brain. The obtained results indicate that in both hippocampal parts the cellular density of the CA1 pyramidal neurons is lower in human than in tenrec. However, while the human pyramidal cell density is higher in the septal hippocampal part than in the temporal one, in the tenrec the density of these cells is higher in the temporal part. The dendritic tree of the CA1 pyramidal cells, more developed in the septal than in temporal hippocampus in both species studied, is in general more complex in the human hippocampus. The basal and the apical dendritic systems exhibit species related morphometric differences, while dendrites of different orders exhibit differences in their number and length, and in their spine density. Finally, in both species, as well as hippocampal parts and dendritic systems, changes of dendritic morphometric features along ascending dendritic orders fluctuate in a similar way, as do the number of dendritic branch points in relation to the distance from the neuron soma. PMID- 18511021 TI - Orexin-2 receptors inhibit primary afferent fiber-evoked responses of ventral roots in the neonatal rat isolated spinal cord. AB - Orexin-A and orexin-B are hypothalamic peptides. Orexin-A binds equally to both orexin-1 and orexin-2 receptors but orexin-B has a preferential affinity for orexin-2 receptor. In the spinal dorsal horn, orexins have been shown to be concentrated in the superficial laminae. In the present study, the authors examined the effect of spinally applied orexin-A and orexin-B on the primary afferent fiber-evoked nociceptive reflex in the isolated spinal cord of the neonatal rat. In the isolated spinal cord preparation from 0-3day old rats, single-shock stimulation of a dorsal root (L3-L5) at a strength which can activate C-fibers induced a slow depolarizing response lasting about 30s (slow ventral root potential: slow VRP) in the ipsilateral ventral root of the same segment. Bath application of orexin-A and orexin-B inhibited the slow VRP in a concentration-dependent manner. Bath application of SB-334867, a selective orexin 1 receptor antagonist, had no effect on the depressant effect of orexin-A on slow VRP. Bath application of [Ala11,d-Leu15]-orexin B, a selective orexin-2 receptor agonist, depressed the slow VRP. Both orexin-A and orexin-B depressed the level of temporal summation of synaptic activity evoked by 20 repetitive stimulations of the dorsal root. These data suggest that orexin-2 receptor, but not orexin-1 receptor, may play an inhibitory role in nociceptive transmission in the neonatal rat spinal cord. PMID- 18511022 TI - Peripheral neuritis and increased spinal cord neurochemicals are induced in a model of repetitive motion injury with low force and repetition exposure. AB - Performance of high repetition tasks with or without force is associated with peripheral tissue inflammation, decreased nerve function and motor dysfunction. Here, we examined whether a low repetition task with negligible force (LRNF) produces fewer tissue and behavioral pathologies than previously observed with high repetition tasks using our rat model of repetitive motion injury (RMI). Thirty-seven rats were randomized into control or LRNF groups, the latter reaching and grasping a 45 mg food pellet at a rate of 3 reaches/min. This task was performed in 4, 0.5 5 h sessions with 1.5 5 h rest periods for 3 days/week for up to 12 weeks. Examination of distal median nerve, forelimb flexor tendons and bones for ED1-positive cells (macrophages and osteoclasts) revealed increases in nerve and bone in week 12. The nerve also contained increased TNF-alpha expressing cells in week 12. Examination of spinal cord dorsal horns revealed increased immunoexpression of Substance P in week 8 and neurokinin-1 in weeks 8 and 12 in the superficial lamina. Motor behavioral analyses showed no changes in reach rate across weeks, slightly reduced task duration (a measurement of voluntary task participation) in week 12, but significantly increased extra arm movement reversals during reaching in week 8. These extra movement reversals were corrections for missed food pellets during a reach. Thus, performance of even a low repetition, negligible force upper extremity task for 3 months can induce mild peripheral tissue inflammation, neurochemical increases in spinal cord dorsal horns, and declines in fine motor control. PMID- 18511023 TI - Sleep deprivation alters functioning within the neural network underlying the covert orienting of attention. AB - One function of spatial attention is to enable goal-directed interactions with the environment through the allocation of neural resources to motivationally relevant parts of space. Studies have shown that responses are enhanced when spatial attention is predictively biased towards locations where significant events are expected to occur. Previous studies suggest that the ability to bias attention predictively is related to posterior cingulate cortex (PCC) activation [Small, D.M., et al., 2003. The posterior cingulate and medial prefrontal cortex mediate the anticipatory allocation of spatial attention. Neuroimage 18, 633-41]. Sleep deprivation (SD) impairs selective attention and reduces PCC activity [Thomas, M., et al., 2000. Neural basis of alertness and cognitive performance impairments during sleepiness. I. Effects of 24 h of sleep deprivation on waking human regional brain activity. J. Sleep Res. 9, 335-352]. Based on these findings, we hypothesized that SD would affect PCC function and alter the ability to predictively allocate spatial attention. Seven healthy, young adults underwent functional magnetic resonance imaging (fMRI) following normal rest and 34-36 h of SD while performing a task in which attention was shifted in response to peripheral targets preceded by spatially informative (valid), misleading (invalid), or uninformative (neutral) cues. When rested, but not when sleep deprived, subjects responded more quickly to targets that followed valid cues than those after neutral or invalid cues. Brain activity during validly cued trials with a reaction time benefit was compared to activity in trials with no benefit. PCC activation was greater during trials with a reaction time benefit following normal rest. In contrast, following SD, reaction time benefits were associated with activation in the left intraparietal sulcus, a region associated with receptivity to stimuli at unexpected locations. These changes may render sleep-deprived individuals less able to anticipate the locations of upcoming events, and more susceptible to distraction by stimuli at irrelevant locations. PMID- 18511024 TI - Neurotrophin-induced upregulation of p75NTR via a protein kinase C-delta dependent mechanism. AB - Neurotrophins exert their biological effects via p75NTR and Trk receptors. Functional interplay between these two receptors has been widely explored with respect to p75NTR enhancing the activation and signalling of Trk, but few studies address the bidirectional aspects. We have previously demonstrated that the expression of p75NTR can be differentially modulated by different Trk receptor mutations. Here we investigate the mechanism of Nerve Growth Factor (NGF)-induced upregulation of p75NTR expression. We utilize pharmacological inhibition to investigate the role of various TrkA-associated signalling intermediates in this regulatory cascade. Notably, the inhibition of phospholipase C-gamma (PLC-gamma) using U73122, prevented the NGF-induced upregulation of p75NTR protein and mRNA. The inhibition of protein kinase C-delta (PKC-delta) activation by rottlerin, a selective PKC-delta inhibitor, and by small interfering RNA (siRNA) directed against PKC-delta also inhibited this NGF-induced upregulation. Finally, we also show that in cerebellar granule neurons, BDNF acting via TrkB increases p75NTR expression in a PKC-delta dependent manner. These results indicate the importance of Trk-dependent PLC-gamma and PKC-delta activation for downstream regulation of p75NTR protein expression in response to neurotrophin stimulation, a process that has implications to the survival and growth of the developing nervous system. PMID- 18511025 TI - The effects of rewarding ventral tegmental area stimulation and environmental enrichment on lipopolysaccharide-induced sickness behavior and cytokine expression in female rats. AB - Responding for rewarding brain stimulation has been used to track hedonic status in animals. In addition to neurochemical alterations, stimulation of the lateral hypothalamus or ventral tegmentum has been shown to influence immunological processes, including elevation of peripheral natural killer cell activity. In the present study, we examined whether ventral tegmental area (VTA) stimulation or environmental enrichment altered the severity of lipopolysaccharide (LPS)-induced sickness behaviors and the provocation of cytokine expression induced by the endotoxin. Accordingly, rats received either trials of brain stimulation reward or exposure to an enriched environment and subsequently challenged with 150 ug/kg i.p. of LPS. Groups receiving LPS and saline injections without further manipulation were also included. Using the real-time RT-PCR and a multiplex bead assay, mRNA and protein levels for several cytokines and their receptors were determined to evaluate how these may vary as a consequence of reward. Both brain stimulation and environmental enrichment similarly diminished sickness behaviors associated with the endotoxin. Receptor gene levels were generally stable across groups. Levels of IL-6 within the VTA were increased in the group receiving LPS challenge alone and environmental enrichment was associated with modestly reduced IL-6 levels within this brain region. Taken together, these data suggest that rewarding brain stimulation and environmental enrichment buffer against malaise provoked by endotoxin challenge. Moreover, IL-6 expression within the VTA may influence the development of sickness behavior following inflammatory stimuli. PMID- 18511026 TI - Calculation of viscometric constants, hydrodynamic volume, polymer-solvent interaction parameter, and expansion factor for three polysaccharides with different chain conformations. AB - The viscometric constants, K and a, for three polysaccharides: hydroxyethylcellulose (HEC); hydroxypropylcellulose (HPC); and chitosan, were calculated at 30 degrees C using intrinsic viscosity, [eta] and molecular weight (M(n), M(w), M(z)) data. The polydispersity correction factor, q(MHS), and hydrodynamic volume for each polymer sample were also calculated. The value of q(MHS) for the polymer samples was taken into account in the calculation of the viscometric constants. The polymer-solvent interaction parameters for the three polysaccharides were estimated by both semiempirical and numerical methods using intrinsic viscosity and molecular weight data. Hydrodynamic expansion factors were also estimated using the latter data. The quality of the solvents for the three polymers was compared using exponent a, polymer-solvent interaction parameter, and expansion factor data. This study resulted in the following constants for: The values of 0.60, 1.08, and 0.885 for exponents a indicate that HEC, HPC, and chitosan behave as a flexible random coil, linear and extended conformations, respectively. The values of exponents a for the three polysaccharides appear to be inversely related to their K values. The results of the expansion factor were consistent with the results of exponent a and polymer solvent interaction parameters. PMID- 18511027 TI - Synthesis of fluorescent alkyl lactoside derivatives. AB - Fluorescent-labeled alkyl lactoside (NBD-alkyl lactoside) derivatives were synthesized by the reaction of NBD alkyl alcohol with heptaacetyllactosyl trichloroacetimidate in the presence of BF(3).Et(2)O. PMID- 18511028 TI - Cytokine production by M-CSF- and GM-CSF-induced mouse bone marrow-derived macrophages upon coculturing with late apoptotic cells. AB - Recently, we found that resident peritoneal macrophages produce MIP-2, one of the major chemokines for neutrophils, upon coculturing with late apoptotic cells, and that intraperitoneal injection of late apoptotic cells into the peritoneal cavity causes neutrophil infiltration via MIP-2. It is not known, however, whether or not macrophages are heterogeneous in such MIP-2 production. In this study, we examined changes in the surface phenotype during the differentiation of bone marrow cells into macrophages due to M-CSF and GM-CSF, and then examined the production of cytokines, namely IL-12 p40, MIP-2, IL-10, and TGF-beta, following phagocytosis of late apoptotic cells with these macrophages or LPS stimulation of these macrophages. GM-CSF and M-CSF induced macrophage populations with distinct but overlapping cell surface phenotype. Although these macrophages phagocytosed late apoptotic cells to a similar extent, they produced either IL-12 p40 or IL 10, whereas they produced MIP-2 to a similar extent after the coculture, raising the possibility that late apoptotic cells may induce neutrophil infiltration in any organs, such as the liver and lungs, where the macrophages are differentiated by either M-CSF or GM-CSF, respectively. PMID- 18511029 TI - Antioxidant activity and inhibition of aflatoxin B1-, nifuroxazide-, and sodium azide-induced mutagenicity by extracts from Rhamnus alaternus L. AB - The effect of extracts obtained from Rhamnus alaternus L. leaves on genotoxicity and SOS response induced by aflatoxin B(1) (10 microg/assay) as well as nifuroxazide (20 microg/assay) was investigated in a bacterial assay system, i.e., the SOS chromotest with Escherichia coli PQ37. The evaluation of the mutagenic and antimutagenic actions of the same extracts against the sodium azide (1.5 microg/plate)-induced mutagenicity was assayed using the Salmonella typhimurium assay system. The R. alaternus tested extracts exhibited no genotoxicity either with or without the external S9 activation mixture. However, all the extracts, particularly aqueous extract (A) and its chloroformic fraction (A(2)) significantly decreased the genotoxicity induced by aflatoxin B(1) and nifuroxazide. Moreover, the different extracts showed no mutagenicity when tested with Salmonella typhimurium strains TA1535 and TA1538 either with or without the S9 mix. Aqueous extract as well as its A(2) fraction exhibited the highest level of protection towards the direct mutagen, sodium azide-induced response in TA1535 strain with mutagenicity inhibition percentages of 83.6% and 91.4%, respectively, at a dose of 250 microg/plate. The results obtained by the Ames test assay confirm those of SOS chromotest. These same active extracts exhibited high xanthine oxidase (XOD) inhibiting with respective IC(50) values of 208 and 137 microg/ml, and superoxide anion-scavenging effects (IC(50) values of 132 and 117 microg/ml) when tested in the XOD enzymatic assay system. Our findings emphasize the potential of R. alaternus to prevent mutations and also its antioxidant effect. PMID- 18511030 TI - Notch signaling during larval and juvenile development in the polychaete annelid Capitella sp. I. AB - Notch signaling is involved in a large range of developmental processes, and has been functionally implicated in body plan segmentation in two of the three diverse segmented taxa, the vertebrates and arthropods. Here we investigate expression of Notch, Delta, and hes gene homologues during larval and juvenile development in the polychaete annelid Capitella sp. I., a member of the third group of segmented animals. During larval stages, CapI-Notch, CapI-Delta, CapI hes2, and CapI-hes3 transcripts are initially detected in broad ectodermal domains in future segments as well as in the brain and foregut; later, CapI Notch, CapI-Delta, and CapI-hes2 transcripts are detected in the presumptive chaetal sacs. In contrast, CapI-hes1 has a segmentally reiterated pattern in a restricted region of the mesoderm in each presumptive segment. CapI-Notch, CapI Delta, CapI-hes2, and CapI-hes3 and CapI-hes1 are all expressed in the terminal growth zone that generates post-metamorphic segments, however, CapI-hes1 has a non-overlapping complementary expression pattern to that of CapI-Notch and CapI Delta. CapI-Delta and CapI-Notch transcripts are localized to already formed segments, with posterior boundaries that correlate with the posterior boundary of the nascent segment, while CapI-hes1 lies posterior to CapI-Notch and CapI-Delta. The localization of CapI-Notch, CapI-Delta, and CapI-hes transcripts correlate with areas of rapid cell proliferation in Capitella, which include the brain, foregut, and terminal growth zone. PMID- 18511031 TI - Arsenic in drinking water and lung cancer: a systematic review. AB - Exposure to inorganic arsenic via drinking water is a growing public health concern. We conducted a systematic review of the literature examining the association between arsenic in drinking water and the risk of lung cancer in humans. Towards this aim, we searched electronic databases for articles published through April 2006. Nine ecological studies, two case-control studies, and six cohort studies were identified. The majority of the studies were conducted in areas of high arsenic exposure (100 microg/L) such as southwestern Taiwan, the Niigata Prefecture, Japan, and Northern Chile. Most of the studies reported markedly higher risks of lung cancer mortality or incidence in high arsenic areas compared to the general population or a low arsenic exposed reference group. The quality assessment showed that, among the studies identified, only four assessed arsenic exposure at the individual level. Further, only one of the ecological studies presented results adjusted for potential confounders other than age; of the cohort and case-control studies, only one-half adjusted for cigarette smoking status in the analysis. Despite these methodologic limitations, the consistent observation of strong, statistically significant associations from different study designs carried out in different regions provide support for a causal association between ingesting drinking water with high concentrations of arsenic and lung cancer. The lung cancer risk at lower exposure concentrations remains uncertain. PMID- 18511032 TI - Effects of angiotensin II and its metabolites in the rat coronary vascular bed: is angiotensin III the preferred ligand of the angiotensin AT2 receptor? AB - Aminopeptidases metabolize angiotensin II to angiotensin-(2-8) (=angiotensin III) and angiotensin-(3-8) (=angiotensin IV), and carboxypeptidases generate angiotensin-(1-7) from angiotensin I and II. Angiotensin-converting enzyme (ACE) inhibitors and/or angiotensin II type 1 (AT1) receptor blockers affect the concentrations of these metabolites, and they may thus contribute to the beneficial effects of these drugs, possibly through stimulation of non-classical angiotensin AT receptors. Here, we investigated the effects of angiotensin II, angiotensin III, angiotensin IV and angiotensin-(1-7) in the rat coronary vascular bed, with or without angiotensin AT1 - or angiotensin II type 2 (AT2) receptor blockade. Results were compared to those in rat iliac arteries and abdominal aortas. Angiotensin II, angiotensin III and angiotensin IV constricted coronary arteries via angiotensin AT1 receptor stimulation, angiotensin III and angiotensin IV being approximately 20- and approximately 8000-fold less potent than angiotensin II. The angiotensin AT2 receptor antagonist PD123319 greatly enhanced the constrictor effects of angiotensin III, starting at angiotensin III concentrations in the low nanomolar range. PD123319 enhanced the angiotensin II induced constriction at submicromolar angiotensin II concentrations only. Angiotensin-(1-7) exerted no effects in the coronary circulation, although, at micromolar concentrations, it blocked angiotensin AT1 receptor-induced constriction. Angiotensin AT2 receptor-mediated relaxation did not occur in iliac arteries and abdominal aortas, and the constrictor effects of the angiotensin metabolites in these vessels were identical to those in the coronary vascular bed. In conclusion, angiotensin AT2 receptor activation in the rat coronary vascular bed results in vasodilation, and angiotensin III rather than angiotensin II is the preferred endogenous agonist of these receptors. Angiotensin II, angiotensin III, angiotensin IV and angiotensin-(1-7) do not exert effects through non-classical angiotensin AT receptors in the rat coronary vascular bed, iliac artery or aorta. PMID- 18511033 TI - Synergy between a NR2B receptor antagonist DHQ and 3-methyl-gabapentin in mice with neuropathic pain. AB - The synergistic effect of a selective NR2B NMDA receptor antagonist, (-)-(R)-6-{2 [4-(3-Fluorophenyl)-4-hydroxy-1-piperidinyl]-1-hydroxyethyl-3,4-dihydro-2(1H) quinolinone (DHQ), and a alpha 2 delta ligand, 3-methyl-gabapentin (3M-GBP), was investigated in the mouse partial sciatic nerve model. The interaction was observed after administration of DHQ and 3M-GBP combination at fixed dose ratios of 1:10 and 1:30 and the dose-response curves shifted approximately 13- and 17 fold leftward, respectively, from the theoretical additive values. However, a fixed dose ratio of 1:50 resulted only in an additive effect. These results indicate the synergistic interaction between DHQ and 3M-GBP in this animal model of neuropathic pain. PMID- 18511034 TI - Actions of novel agonists, antagonists and antipsychotic agents at recombinant rat 5-HT6 receptors: a comparative study of coupling to G alpha s. AB - Though 5-HT6 receptors are targets for the treatment of schizophrenia and other psychiatric disorders, the influence of drugs upon signal transduction has not been extensively characterized. Herein, we employed a Scintillation Proximity Assay (SPA)/antibody-immunocapture procedure of coupling to G alpha s to evaluate the interaction of a broad range of novel agonists, antagonists and antipsychotics at rat 5-HT(6) receptors stably expressed in HEK293 cells. Serotonin (pEC(50), 7.7) increased [35S]GTP gamma S binding to G alpha s by ca 2 fold without affecting binding to Gi/o or Gq. LSD (9.2), 5-MeODMT (7.9), 5-CT (7.0) and tryptamine (6.1) were likewise full agonists. In contrast, the novel sulfonyl derivatives, WAY181,187 (9.1) and WAY208,466 (7.8), behaved as partial agonists and attenuated the actions of 5-HT. SB271,046 and SB258,585 abolished activation of G alpha s by 5-HT with pKb values of 10.2 and 9.9, respectively, actions mimicked by the novel antagonist, SB399,885 (10.9). SB271,046 likewise blocked partial agonist properties of WAY181,187 and WAY208,466 with pKb values of 9.8 and 9.0, respectively. 5-HT-stimulated [35S]GTP gamma S binding to G alpha s was antagonised by various antipsychotics including olanzapine (7.8), asenapine (9.1) and SB737,050 (7.8), whereas aripiprazole and bifeprunox were inactive. Further, antagonist properties of clozapine (8.0) were mimicked by its major metabolite, N-desmethylclozapine (7.9). In conclusion, the novel ligands, WAY208,466 and WAY181,187, behaved as partial agonists at 5-HT6 receptors coupled to G alpha s, while SB399,885 was a potent antagonist. Though 5-HT6 receptor blockade is not indispensable for therapeutic efficacy, it may well play a role in the functional actions of certain antipsychotic agents. PMID- 18511035 TI - Amyloid beta peptide-activated signal pathways in human platelets. AB - Amyloid beta peptide (amyloid-beta), which accumulates in the cerebral microvessels in an age-dependent manner, plays a key role in the pathogenesis of cerebral amyloid angiopathy. Platelets are an important cellular element in vasculopathy of various causes. Amyloid-beta may activate or potentiate platelet aggregation. The present study explored the signaling events that underlie amyloid-beta activation of platelet aggregation. Platelet aggregometry, immunoblotting and assays to detect activated cellular events were applied to examine the signaling processes of amyloid-beta activation of platelets. Exogenous amyloid-beta (1-2 microM) potentiated platelet aggregation caused by collagen and other agonists. At higher concentrations (5-10 microM), amyloid-beta induced platelet aggregation which was accompanied by an increase in thromboxane A2 (TxA2) formation. These amyloid-beta actions on platelets were causally related to amyloid-beta activation of p38 mitogen-activated protein kinase (MAPK). Inhibitors of p38 MAPK and its upstream signaling pathways including proteinase-activated receptor 1 (PAR1), Ras, phosphoinositide 3-kinase (PI3 kinase), or Akt, but not extracellular signal-regulated kinase 2 (ERK2)/c-Jun N terminal kinase 1 (JNK1), blocked amyloid-beta-induced platelet activation. These findings suggest that the p38 MAPK, but not ERK2 or JNK1 pathway, is specifically activated in amyloid-beta-induced platelet aggregation with the following signaling pathway: PAR1 --> Ras/Raf --> PI3-kinase --> Akt --> p38 MAPK --> cytosolic phospholipase A2 (cPLA2)--> TxA2. In conclusion, this study demonstrates amyloid-beta activation of a p38 MAPK signaling pathway in platelets leading to aggregation. Further studies are needed to define the specific role of amyloid-beta activation of platelets in the pathogenesis of vasculopathy including cerebral amyloid angiopathy. PMID- 18511036 TI - Characterization of the unique regulatory mechanisms of phorbol ester-induced polymorphonuclear leukocyte spreading in an acidified environment. AB - In vitro studies have shown that acidic conditions impair spreading of polymorphonuclear leukocytes, which is prerequisite for activation of microbicidal functions. However, the mechanisms by which pH affects polymorphonuclear leukocytes functions remain obscure. Moreover, in vitro observations seem to contradict the fact that an acidic microenvironment often prevails at sites of inflammation where polymorphonuclear leukocytes must function for host defense. In the present study, we found three peculiar characteristics of porcine polymorphonuclear leukocyte that had been induced to spread over fibrinogen-coated surfaces by phorbol 12-myristate 13-acetate (PMA) in acidified medium. First, the PMA-induced spreading at acidic pH, but not at neutral/alkaline pH, was dependent on extracellular Ca2+. Second, the spreading at acidic pH was independent of protein kinase C (PKC), whereas that at neutral/alkaline pH was strictly PKC-dependent. Finally, the spreading at acidic pH, but not at neutral/alkaline pH, was suppressed by H2O2 produced by activated NADPH oxidase or added exogenously. As a result, polymorphonuclear leukocyte spreading at acidic pH peaked at 30 min after PMA stimulation, and declined thereafter because of negative regulation triggered by accumulated H2O2, whereas that at neutral/alkaline pH was stable for at least 90 min. The NADPH oxidase inhibitor diphenyleneiodonium or the H2O2-degradation enzyme catalase consistently stabilized the spreading at acidic pH. We conclude that PMA stimulated polymorphonuclear leukocytes spread in an acidic environment through a mechanism different from that under neutral/alkaline conditions. This H2O2 mediated negative regulation system in an acidic environment may be crucial for avoiding tissue-damaging inflammatory actions of accumulated polymorphonuclear leukocytes in vivo. PMID- 18511037 TI - Heat stress alters G-protein coupled receptor-mediated function and endothelium dependent relaxation in rat mesenteric artery. AB - Heat stress has been demonstrated to have strong cardiovascular effects. However, the underlying mechanism-mediated cardiovascular effects are still not fully understood. The present study was designed to examine if heat stress alters vascular G-protein coupled receptor-mediated vasomotion and endothelium function in rat mesenteric artery. Rats were divided into two groups, heat stress rats and control. The G-protein coupled receptors of endothelin type B (ETB) receptor-, endothelin type A (ETA) receptor-, 5-hydroxytryptamine (5-HT) receptor-, calcitonin gene-related peptide (CGRP) receptor-, alpha-adrenoceptor-mediated vosoactivity and endothelium-dependent relaxation on rat mesenteric artery ring segments were monitored by a myograph system. The plasma level of CGRP was determined by radioimmunological assay. Compared with control arterial segments, the contractile response curves of sarafotoxin 6c, a selective ETB receptor agonist and 5-HT in the arterial segments from heat stress rats were shifted towards left. An increased maximum contraction (Emax) induced by sarafotoxin 6c, but not 5-HT, was seen in the arterial segments from heat stress rats. CGRP induced relaxation in endothelium-denuded arterial segments from heat stress rats was enhanced. The relaxation in endothelium-intact arterial segments induced by acetylcholine was significantly decreased in heat stress rats. In addition, the plasma concentration of CGRP was increased in heat stress rats. The endothelium dependent relaxation was characterized and shown there was a decrease in nitric oxide and endothelium-derived hyperpolarizing factor-mediated relaxation in the arterial segments from heat stress rats. In conclusion, heat stress induces an enhanced vascular endothelin ETB-, 5-HT-receptors-mediated contraction, an enhanced CGRP-receptor-induced relaxation and damage to endothelium-dependent relaxation. PMID- 18511038 TI - Baicalein protects chicken embryonic cardiomyocyte against hypoxia-reoxygenation injury via mu- and delta- but not kappa-opioid receptor signaling. AB - Baicalein, a pure compound derived from Scutellaria baicalensis Georgi, protected cells from lethal damage in an ischemia-reperfusion model. This study was aimed to investigate the role of opioid receptors in mediating cardioprotection by baicalein against hypoxia-reoxygenation injury. By using chick cardiomyocyte as in vitro model, baicalein was added to the perfusate during 1 h-hypoxia followed by 1 h-reoxygenation. Cell viability was assessed by propidium iodide uptake, while apoptosis was assessed by TUNEL and Hoechst 33342 staining. The expression of opioid receptors mRNA in chicken embryonic myocardium was determined by RT PCR. Opioid receptor antagonists, protein kinase C inhibitors, and KATP channel blockers were used to determine the presumed signal transduction pathways. The results showed that baicalein (0.1 approximately 5 microM) concentration dependently reduced hypoxia-reoxygenation-induced myocardial death and apoptosis. The cardioprotective effect of baicalein (1 microM) was blocked by pretreatment of nonspecific opioid receptor antagonist (naloxone), opioid mu-receptor (beta funaltrexamine) and delta-receptor (7-Benzylidenenaltrexone) antagonists, protein kinase C inhibitors (H7 and chelerythrine), and KATP channel blockers (glibenclamide and 5-hydroxydecanoate). Finally, RT-PCR analysis successfully demonstrated the presence of opioid receptors mRNA in chicken embryonic cardiomyocytes. We conclude that the cardioprotective effect of baicalein is mediated via mu-, delta- but not kappa-opioid receptor and their related signal transduction pathways, such as protein kinase C and the KATP channel. PMID- 18511039 TI - Role of IL-18 in overt pain-like behaviour in mice. AB - There are evidences that targeting IL-18 might be beneficial to inhibit inflammatory symptoms, including hypernociception (decrease in nociceptive threshold). The mechanism of IL-18 mechanical hypernociception depends on endothelin in rats and mice. However, the role of IL-18 in overt pain-like behaviour remains undetermined. Therefore, we addressed the role of IL-18 in writhing response induced by intraperitoneal (i.p.) injection of phenyl-p benzoquinone (PBQ) and acetic acid in mice. Firstly, it was detected that PBQ and acetic acid i.p. injection induced a dose-dependent number of writhes in Balb/c mice. Subsequently, it was observed that the PBQ - but not the acetic acid induced writhes were diminished in IL-18 deficient ((-/-)) mice. Therefore, considering that IFN-gamma, endothelin and prostanoids mediate IL-18-induced mechanical hypernociception, we also investigated the role of these mediators in the same model of writhing response in which IL-18 participates. It was noticed that PBQ-induced writhes were diminished in IFN-gamma(-/-) mice and by the treatment with bosentan (mixed endothelin ETA/ETB receptor antagonist), BQ 123 (cyclo[DTrp-DAsp-Pro-DVal-Leu], selective endothelin ETA receptor antagonist), BQ 788 (N-cys-2,6 dimethylpiperidinocarbonyl-l-methylleucyl-d-1-methoxycarboyl-d norleucine, selective endothelin ETB receptor antagonist) or indomethacin (cycloxigenase inhibitor). Thus, IL-18, IFN-gamma, endothelin acting on endothelin ETA and ETB receptors, and prostanoids mediate PBQ-induced writhing response in mice. To conclude, these results further advance the understanding of the physiopathology of overt pain-like behaviour, and suggest for the first time a role for IL-18 in writhing response in mice. PMID- 18511040 TI - Early effects of tocilizumab on bone and bone marrow lesions in a collagen induced arthritis monkey model. AB - To understand the contribution of IL-6/IL-6R to subchondral bone and bone marrow abnormality in RA patients and the effects of tocilizumab on those abnormalities, we evaluated early change in a collagen-induced arthritis (CIA) monkey model with or without a single administration of tocilizumab. Six CIA cynomolgus monkeys received tocilizumab and 3 CIA monkeys received vehicle only. Their interphalangeal joints were analyzed using HE, silver impregnation (SI), or immunohistochemistry (RANKL) staining. The number of osteoclasts increased in the arthritis control but was suppressed in the tocilizumab-treated animals. Osteoblast/stromal cells of the arthritis control monkeys were of monolayer, while in the tocilizumab-treated monkeys, the cells were multi-layer or differentiated osteoblasts, and the meshwork of the reticulum fibers showed recovery in the SI. Hematopoietic marrow was replaced by interstitial fluid and reticulum fibers were eliminated in the arthritic model but showed recovery in the tocilizumab-treated animals. RANKL showed overproduction with arthritis and suppressed with tocilizumab treatment. The evidence indicates that IL-6/IL-6R is involved in subchondral bone and bone marrow change in RA patients. Tocilizumab treatment recovered changes in the CIA monkeys as a result of the co differentiation between the osteoclasts and the osteoblast/stramal cells, at least partially through the suppression of RANKL overproduction. PMID- 18511041 TI - Remote activation of microglia and pro-inflammatory cytokines predict the onset and severity of below-level neuropathic pain after spinal cord injury in rats. AB - Spinal cord injury (SCI) impairs sensory systems causing chronic allodynia. Mechanisms underlying neuropathic pain have been more extensively studied following peripheral nerve injury (PNI) than after central trauma. Microglial activation, pro-inflammatory cytokine production and activation of p38 MAP kinase pathways may induce at-level allodynia following PNI. We investigated whether midthoracic SCI elicits similar behavioral and cellular responses below the level of injury (lumbar spinal cord; L5). Importantly, we show that anatomical connections between L5 and supraspinal centers remain intact after moderate SCI allowing direct comparison to a well-established model of peripheral nerve injury. We found that SCI elicits below-level allodynia of similar magnitude to at-level pain caused by a peripheral nerve injury. Moreover, the presence of robust microglial activation in L5 cord predicted allodynia in 86% of rats. Also increased phosphorylation of p38 MAP kinase occurred in the L5 dorsal horn of allodynic rats. For below-level allodynia after SCI, TNF-alpha and IL-1beta increased in the L5 dorsal horn by 7 dpo and returned to baseline by 35 dpo. Interestingly, IL-6 remains at normal levels early after SCI and increases at chronic time points. Increased levels of pro-inflammatory cytokines also occurred in the thalamus after SCI-induced allodynia. These data suggest that remote microglial activation is pivotal in the development and maintenance of below level allodynia after SCI. Fractalkine, a known activator of microglia, and astrocytes were not primary modulators of below-level pain. Although the mechanisms of remote microglial activation are unknown, this response may be a viable target for limiting or preventing neuropathic pain after SCI in humans. PMID- 18511042 TI - Late expression of Na+/H+ exchanger 1 (NHE1) and neuroprotective effects of NHE inhibitor in the gerbil hippocampal CA1 region induced by transient ischemia. AB - Although acidosis may be involved in neuronal death, the participation of Na(+)/H(+) exchanger (NHE) in delayed neuronal death in the hippocampal CA1 region induced by transient forebrain ischemia has not been well established. In the present study, we investigated the chronological alterations of NHE1 in the hippocampal CA1 region using a gerbil model after ischemia/reperfusion. In the sham-operated group, NHE1 immunoreactivity was weakly detected in the CA1 region. Two and 3 days after ischemia/reperfusion, NHE1 immunoreactivity was observed in glial components, not in neurons, in the CA1 region. Four days after ischemia/reperfusion, NHE1 immunoreactivity was markedly increased in CA1 pyramidal neurons as well as glial cells. These glial cells were identified as astrocytes based on double immunofluorescence staining. Western blot analysis also showed that NHE protein level in the CA1 region began to increase 2 days after ischemia/reperfusion. The treatment of 10 mg/kg 5-(N-ethyl-N-isopropyl) amiloride, a NHE inhibitor, significantly reduced the ischemia-induced hyperactivity 1 day after ischemia/reperfusion. In addition, NHE inhibitor potently protected CA1 pyramidal neurons from ischemic damage, and NHE inhibitor attenuated the activation of astrocytes and microglia in the ischemic CA1 region. In addition, NHE inhibitor treatment blocked Na(+)/Ca(2+) exchanger 1 immunoreactivity in the CA1 region after transient forebrain ischemia. These results suggest that NHE1 may play a role in the delayed death, and the treatment with NHE inhibitor protects neurons from ischemic damage. PMID- 18511043 TI - Neurotrophin-3 deficient Schwann cells impair nerve regeneration. AB - Neurotrophin 3 (NT-3) is an important autocrine factor supporting Schwann cell (SC) survival and differentiation in the absence of axons. Prior studies have failed to define the explicit role of SC versus axon in NT-3 deficiency in relation to nerve regeneration and associated remyelination. In the paradigm we studied, using NT-3 heterozygous (NT3(+/-)) knockout mice capable of survival into adult-life, the experimental design provided a model uniquely capable of differentiating SC/axon influences. In these studies we first identified a defect in nerve regeneration characterized by fewer SCs in the regenerating nerve fibers of crushed sciatic nerves of NT3(+/-) mice. Subsequent experiments differentiated SC versus axonal influences as the culprit in defective nerve regeneration using sciatic nerve transplant paradigms. Results show an impairment in nerve regeneration in NT3(+/-) mice with a retardation of the myelination process, and this defect is associated with decreased SC survival and an increase in the neurofilament packing density of regenerating axons. These observations indicate that NT3(+/-) status of the SCs, but not of the axons, is responsible for impaired nerve regeneration and that NT-3 is essential for SC survival in early stages of regeneration-associated myelination in the adult peripheral nerve. PMID- 18511044 TI - Parkinson patient fibroblasts show increased alpha-synuclein expression. AB - Parkinson's disease (PD) is a neurodegenerative movement disorder of advanced age with largely unknown etiology, but well documented tissue damage from oxidative stress. Increased alpha-synuclein (SNCA) expression is known to cause a rare form of PD, early-onset autosomal dominant PARK4. We have previously shown that loss of-function mutations of the mitochondrial kinase PINK1 which cause the early onset recessive PARK6 variant result in oxidative damage in patient fibroblasts. We now investigated the molecular chain of events from mitochondrial dysfunction to cell death which is largely unknown. Primary skin fibroblast cultures from patients were analysed for gene expression anomalies. In G309D-PINK1 patient fibroblasts, mainly genes regulated by oxidative stress, as well as genes encoding synaptic proteins such as SNCA showed altered expression. The induction of SNCA was also observed in control fibroblasts with knock-down of PINK1. The induction of SNCA expression was found to constitute a specific disease biomarker in sporadic PD patient fibroblasts. To understand the mechanism of this induction, we exposed control fibroblasts to oxidative, proteasomal and endoplasmic reticulum stress and were able to trigger the SNCA expression upregulation. Our data indicate that loss-of-function of PINK1 leads to enhanced alpha-synuclein expression and altered cell-cell contact. Alpha-synuclein induction might represent a common event for different variants of PD as well as a PD-specific trigger of neurodegeneration. We propose that the expression changes described might potentially serve as biomarkers that allow objective PD patient diagnosis in an accessible, peripheral tissue. PMID- 18511045 TI - Xenografts of expanded primate olfactory ensheathing glia support transient behavioral recovery that is independent of serotonergic or corticospinal axonal regeneration in nude rats following spinal cord transection. AB - Transplantation of olfactory ensheathing glial cells (OEG) may improve the outcome from spinal cord injury. Proof-of-principle studies in primates are desirable and the feasibility and efficacy of using in vitro expanded OEG should be tested. An intermediate step between the validation of rodent studies and human clinical trials is to study expanded primate OEG (POEG) xenografts in immunotolerant rodents. In this study the time course to generate purified POEG was evaluated as well as their survival, effect on damaged axons of the corticospinal and serotonergic systems, tissue sparing, and chronic locomotor recovery following transplantation. Fifty-seven nude rats underwent T9/10 spinal cord transection. Thirty-eight rats received POEG, 19 controls were injected with cell medium, and 10 received lentivirally-GFP-transfected POEG. Histological evaluation was conducted at 6 weeks, 8 weeks, 14 weeks and 23-24 weeks. Of these 57 rats, 18 were studied with 5-HT immunostaining, 16 with BDA anterograde CST labeling, and six were used for transmission electron microscopy. In grafted animals, behavioral recovery, sprouting and limited regeneration of 5-HT fibers, and increased numbers of proximal collateral processes but not regeneration of CST fibers was observed. Grafted animals had less cavitation in the spinal cord stumps than controls. Behavioral recovery peaked at three months and then declined. Five POEG-transplanted animals that had shown behavioral recovery underwent retransection and behavioral scores did not change significantly, suggesting that long tract axonal regeneration did not account for the locomotor improvement. At the ultrastructural level presumptive POEG were found to have direct contacts with astrocytes forming the glia limitans, distinct from those formed by Schwann cells. At 6 weeks GFP expression was detected in cells within the lesion site and within nerve roots but did not match the pattern of Hoechst nuclear labeling. At 3.5 months only GFP-positive debris in macrophages could be detected. Transplanted POEG support behavioral recovery via mechanisms that appear to be independent of long tract regeneration. PMID- 18511046 TI - Intracellular and extracellular expression of the major inducible 70kDa heat shock protein in experimental ischemia-reperfusion injury of the spinal cord. AB - Inflammatory responses exacerbate ischemia-reperfusion (IR) injury of spinal cord, although understanding of mediators is incomplete. The major inducible 70kDa heat shock protein (hsp70) is induced by ischemia and extracellular hsp70 (e-hsp70) can modulate inflammatory responses, but there is no published information regarding e-hsp70 levels in the cerebrospinal fluid (CSF) or serum as part of any neurological disease state save trauma. The present work addresses this deficiency by examining e-hsp70 in serum and CSF of dogs in an experimental model of spinal cord IR injury. IR injury of spinal cord caused hind limb paraplegia within 2-3 h that was correlated to lumbosacral poliomalacia with T cell infiltrates at 3 d post-ischemia. In this context, we showed a 5.2-fold elevation of e-hsp70 in CSF that was induced by ischemia and was sustained for the following 3 d observation interval. Plasma e-hsp70 levels were unaffected by IR injury, indicating e-hsp70 release from within the central nervous system. A putative source of this e-hsp70 was ependymal cells in the ischemic penumbra, based upon elevated i-hsp70 levels detected within these cells. Results warrant further investigation of e-hsp70's potential to modulate spinal cord IR injury. PMID- 18511047 TI - Characterization of Leishmania (Leishmania) amazonensis promastigotes resistant to pentamidine. AB - Pentamidine is a second-line agent used in the treatment of leishmaniasis and its mode of action and mechanism of resistance is not well understood. It was previously demonstrated that transfection of promastigotes and amastigotes with the ABC transporter PRP1 gene confers resistance to pentamidine. To further clarify this point, we generated Leishmania amazonensis mutants resistant to pentamidine. Our results indicated that this ABC transporter is not associated with pentamidine resistance in lines generated by drug pressure through amplification or overexpression mechanisms of PRP1 gene. PMID- 18511048 TI - An assessment of computer-assisted personalized sedation: a sedation delivery system to administer propofol for gastrointestinal endoscopy. AB - BACKGROUND: Demand for colonoscopy and EGD procedures is increasing. Impediments to performing these examinations persist. Patients perceive these procedures as unpleasant and painful. The use of suboptimal sedatives results in inefficiency in endoscopy practices. Improving sedation methods utilizing precise control of preferred sedatives may increase patient satisfaction and practice efficiency. OBJECTIVE: Our purpose was to demonstrate the feasibility of computer-assisted personalized sedation (CAPS) for facilitating the precise administration of propofol by endoscopist/nurse teams, achieving minimal to moderate sedation in subjects undergoing routine endoscopies. DESIGN: Open label, single-center studies. SETTING: Endoscopy clinics in Charlottesville, Virginia, and Gent, Belgium. SUBJECTS: Twenty-four adults per center; 12 colonoscopies, 12 EGDs. INTERVENTIONS: Propofol sedation with CAPS by endoscopist/registered nurse care teams. MAIN OUTCOME MEASUREMENTS: Sedation level measured by modified observer's assessment of alertness/sedation (MOAA/S), recovery time measured from endoscope removal until Aldrete >/= 12, dosage of propofol, oxygen saturation, and safety assessments. RESULTS: Subjects responded to mild tactile and verbal stimuli MOAA/S = 5, 4, 3, or 2) 99% of the time. Mean propofol doses in the United States and Belgium were 65.4 and 72.1 mg, respectively. Mean recovery times were 29 and 10 seconds, respectively. Oxygen desaturation occurred in only 6% of subjects. No device-related adverse events occurred. LIMITATION: Open-label design. CONCLUSIONS: Using CAPS, the endoscopist/nurse teams precisely controlled the administration of propofol achieving minimal to moderate sedation in subjects undergoing colonoscopy and EGD procedures. Mean propofol dosage was low and post procedure recovery times were rapid. The device performed well when operated by the endoscopist/nurse team, with no device-related adverse events. PMID- 18511049 TI - A randomized, controlled, double-blind trial of the adjunct use of tegaserod in whole-dose or split-dose polyethylene glycol electrolyte solution for colonoscopy preparation. AB - BACKGROUND: Problems of compliance, quality, and safety of colon preparation regimens have prompted continued investigation with alternative forms of cleansing. OBJECTIVE: To evaluate the efficacy of tegaserod as an adjunct to a polyethylene glycol electrolyte solution (PEG-E), given as a whole dose or split dose, in colonoscopy preparation. DESIGN: Randomized, placebo-controlled, double blind trial. SETTING: A single university-based hospital. PATIENTS: Patients who were undergoing elective colonoscopy. INTERVENTIONS: A 4-arm randomization scheme that compared tegaserod with a placebo, each with whole-dose or split-dose PEG-E preparation. MAIN OUTCOME MEASUREMENTS: Efficacy of colon cleansing was the primary outcome. Secondary outcomes included adherence, tolerability, adverse effects, and patient perceptions of their preparation quality. RESULTS: A total of 382 patients completed the trial. Patients who received the split-dose preparation had significantly better colon cleansing than those who received the whole-dose preparation (88.9% vs 42.6%, P < .001). The addition of tegaserod did not significantly improve the overall colonoscopy preparation quality compared with a placebo. However, there were fewer poor preparations in the whole-dose PEG E group (12.4% vs 1.1%, P = .002, Bonferroni correction removes significance) and more excellent preparations in the split-dose group (53.3% vs 38.3%, P = .035, Bonferroni correction removes significance) in favor of tegaserod. Interobserver and intraobserver variability analysis showed substantial agreement among endoscopists. Adherence was significantly lower in the whole-dose group versus the split-dose PEG-E group (68.8% vs 91%, P < .001), independent of the use of tegaserod. Adverse effects were not different between study groups. LIMITATIONS: A 4-arm randomization and the single-center nature of the study. CONCLUSIONS: Tegaserod has a marginal effect on the quality of colonoscopy preparation when used as an adjuvant to PEG-E. The split-dose PEG-E was superior to the whole-dose PEG-E and resulted in better colon cleansing, adherence, and tolerance. PMID- 18511050 TI - Biological indices, energy reserves, steroid hormones and sexual maturity in the infaunal bivalve Scrobicularia plana from three sites differing by their level of contamination. AB - The aim of this work was to investigate in situ biometric (condition index, hepato-somatic index, gonado-somatic index), biochemical (glycogen, lipids, sexual steroids) and histological (sex, sexual maturation stage) variables in the infaunal bivalve Scrobicularia plana. First, the reproductive cycle of S. plana was assessed by collecting bivalves from a reference site (the bay of Bourgneuf, Fr) in April, May, July, September and November 2005 and in January and March 2006. Then, S. plana were collected at three key periods of their sexual cycle (March 2006, beginning of gametogenesis; June 2006, spawning; and September 2006, spent) in three sites differing by their level of contamination (bay of Bourgneuf, reference site; Loire and Seine estuaries; Fr, impacted sites). The reproductive cycle of S. plana was well defined with a clear spawning period between May and July, sexual repose from November to January. Development of the gonad began in January and ended in September. Sex-ratio was determined during spawning and the influence of sex on biochemical variables was examined. Progesterone, 17beta-estradiol and testosterone levels in the gonad of S. plana were close to those reported in other bivalves. This study is the first to demonstrate in situ influence of site, sex and sexual maturity on energy reserves, and sexual steroids in S. plana. Even if interpretation of results is complex due to interferences between natural and anthropogenic factors, S. plana is a suitable species for estuarine studies and a better understanding of its reproduction will permit to assess impacts of environmental pollutants. PMID- 18511051 TI - Effects of gonadectomy and hormone replacement on a spontaneous novel object recognition task in adult male rats. AB - Recent studies in adult male rats have shown that gonadal hormones influence performance on certain working memory and other types of cognitive tasks that are sensitive to lesions of the medial and/or orbital prefrontal cortices. This study asked whether gonadal hormone modulation of prefrontal cortical function in males also extends to the perirhinal division of the rat prefrontal cortex. Specifically, sham-operated control, gonadectomized, and gonadectomized rats supplemented with testosterone propionate or estradiol were tested on a spontaneous novel object recognition task, a paradigm where performance has been shown to be impaired by perirhinal cortical lesions. Using analyses of variance, regression analyses and post-hoc testing to evaluate group differences, it was found that during both the sample and test trials of the task all four groups spent similar absolute and proportional amounts of time ambulating, rearing, stationary, and exploring the two objects present. All groups also explored each of the two identical objects present during sample trials equally. However, during the test trials, only the control and gonadectomized rats given testosterone showed the expected increase in exploration of the novel objects presented, whereas the gonadectomized and gonadectomized, estradiol-supplemental groups continued to explore the novel and familiar objects equally. That regression analyses also identified significant correlations between low bulbospongiosus muscle weight and impaired novel vs. familiar object discrimination further indicates that gonadectomy in adult male rats adversely affects spontaneous novel object recognition in an androgen-sensitive, estrogen insensitive manner. PMID- 18511052 TI - The value of trauma registries. AB - Trauma registries are databases that document acute care delivered to patients hospitalised with injuries. They are designed to provide information that can be used to improve the efficiency and quality of trauma care. Indeed, the combination of trauma registry data at regional or national levels can produce very large databases that allow unprecedented opportunities for the evaluation of patient outcomes and inter-hospital comparisons. However, the creation and upkeep of trauma registries requires a substantial investment of money, time and effort, data quality is an important challenge and aggregated trauma data sets rarely represent a population-based sample of trauma. In addition, trauma hospitalisations are already routinely documented in administrative hospital discharge databases. The present review aims to provide evidence that trauma registry data can be used to improve the care dispensed to victims of injury in ways that could not be achieved with information from administrative databases alone. In addition, we will define the structure and purpose of contemporary trauma registries, acknowledge their limitations, and discuss possible ways to make them more useful. PMID- 18511053 TI - Bioluminescent imaging of Trypanosoma cruzi infection. AB - Chagas disease, caused by infection with the protozoan parasite Trypanosoma cruzi, is a major public health problem in Central and South America. The pathogenesis of Chagas disease is complex and the natural course of infection is not completely understood. The recent development of bioluminescence imaging technology has facilitated studies of a number of infectious and non-infectious diseases. We developed luminescent T. cruzi to facilitate similar studies of Chagas disease pathogenesis. Luminescent T. cruzi trypomastigotes and amastigotes were imaged in infections of rat myoblast cultures, which demonstrated a clear correlation of photon emission signal strength to the number of parasites used. This was also observed in mice infected with different numbers of luminescent parasites, where a stringent correlation of photon emission to parasite number was observed early at the site of inoculation, followed by dissemination of parasites to different sites over the course of a 25-day infection. Whole animal imaging from ventral, dorsal and lateral perspectives provided clear evidence of parasite dissemination. The tissue distribution of T. cruzi was further determined by imaging heart, spleen, skeletal muscle, lungs, kidneys, liver and intestines ex vivo. These results illustrate the natural dissemination of T. cruzi during infection and unveil a new tool for studying a number of aspects of Chagas disease, including rapid in vitro screening of potential therapeutical agents, roles of parasite and host factors in the outcome of infection, and analysis of differential tissue tropism in various parasite-host strain combinations. PMID- 18511054 TI - Carotid intima-media thickness is increased in patients with spinal cord injury independent of traditional cardiovascular risk factors. PMID- 18511055 TI - Glycation of LDL in non-diabetic people: Small dense LDL is preferentially glycated both in vivo and in vitro. AB - OBJECTIVE: LDL atherogenicity is frequently attributed to oxidative modification, but glycated LDL, which can participate in many of the cellular processes leading to atherosclerosis, generally circulates at higher concentration even in non diabetic people. We tested the hypothesis that small-dense LDL, known to be most closely associated with coronary heart disease, undergoes more glycation than other LDL sub-fractions. METHODS AND RESULTS: The concentration of glycated apolipoprotein B (apo B) was measured in serum, LDL and its sub-fractions from 44 non-diabetic subjects. By ELISA serum glycated apoB concentration was 3.0+/ 1.1mg/dl (mean+/-S.D.) of which 84.6+/-13.6% was in LDL. Of the glycated apo B in LDL 67.8+/-21.9% was in small dense LDL (LDL3; D1.044-1.063g/ml) whereas only 32.2+/-21.9% was in more buoyant LDL subfractions (LDL1 and 2; D1.019-1.044g/ml). The percentage of apo B present in LDL1 and 2 which was glycated was 1.8+/-1.8% whereas in LDL3 it was 17.4+/-18.5% (P<0.001). Furthermore when LDL sub-fractions from non-diabetics (n=29) were incubated with glucose (30-80mmol/l) glycation of apo B in the denser LDL3 subfraction was significantly more pronounced than in less dense LDL subfractions. CONCLUSION: Small-dense LDL is more susceptible to glycation and this may contribute to the atherogenicity of small-dense LDL, even in non-diabetic people. PMID- 18511056 TI - Small arterial elasticity predicts the extent of coronary artery disease: Relationship with serum uric acid. AB - BACKGROUND: Arterial elasticity has been previously linked to atherosclerotic vascular disease states. Serum uric acid level has been recently associated with increased arterial stiffness, but to what extent serum uric acid reflects angiographic coronary artery status and vessel compliance remains to be established. In this study we aimed to evaluate the association of arterial elasticity indexes, serum uric acid and the presence and extent of angiographic coronary artery disease (CAD) in patients with chronic stable angina. METHODS: One hundred and eight consecutive patients attending for elective coronary angiography were investigated. The severity of CAD was expressed using the Gensini score. Quantitative analysis of the arterial elasticity was performed by applanation tonometry. Serum uric acid was measured in all participants. Stepwise multiple linear regression analysis was used to identify the independent correlates of the Gensini score. RESULTS: After adjustment for age, gender, common cardiac risk factors and cardiovascular drugs, small artery elasticity index (SAEI) (p<0.001) and serum uric acid (p<0.001) were independently correlated with the severity of CAD. Stepwise multiple linear regression analysis was also used to identify independent correlates of the SAEI. Serum uric acid emerged as the only independent correlate of SAEI (p<0.001). CONCLUSIONS: SAEI independently reflects the extent of CAD in patients with chronic stable angina. This relationship is chiefly mediated by serum uric acid. Our data add to the growing evidence that serum uric acid may be a marker of arterial stiffness and atherosclerotic burden. PMID- 18511057 TI - Adiponectin reduces lipid accumulation in macrophage foam cells. AB - Adiponectin is one of several, important metabolically active cytokines secreted from adipocytes. Low circulating levels of this adipokine have been associated epidemiologically with obesity, insulin resistance, type II diabetes, and cardiovascular disease. To determine if adiponectin can modulate lipid metabolism in macrophages, we expressed the adiponectin gene in human THP-1 macrophage foam cells using a lentiviral vector expression system and demonstrated that macrophages transduced with the adiponectin gene had decreased lipid accumulation compared with control macrophages transduced with the LacZ gene. Macrophages transduced with the adiponectin gene also exhibited decreased oxidized low density lipoprotein (oxLDL) uptake and increased HDL-mediated cholesterol efflux. Additional studies suggest two potential mechanisms for the reduced lipid accumulation in these adiponectin-transduced macrophage foam cells. The first mechanism involves the PPARgamma and LXR signaling pathways which up-regulate the expression of ABCA1 and promote lipid efflux from these cells. The second mechanism involves decreased lipid uptake and increased lipid hydrolysis which may result from decreased SR-AI and increased SR-BI and HSL gene activities in the transformed macrophage foam cells. We also demonstrated that the expression of two proatherogenic cytokines, MCP-1 and TNFalpha, were decreased in the adiponectin-transduced macrophage foam cells. These results suggest that adiponectin may modulate multiple pathways of lipid metabolism in macrophages. Our studies provide new insights into potential mechanisms of adiponectin mediated alterations in lipid metabolism and macrophage foam cell formation which may impact the development of atherosclerosis. PMID- 18511058 TI - Analysis of brominated flame retardants in styrenic polymers. Comparison of the extraction efficiency of ultrasonication, microwave-assisted extraction and pressurised liquid extraction. AB - The extraction efficiency of pressurised liquid extraction (PLE), microwave assisted extraction (MAE), and ultrasonic-assisted extraction (UAE) under different conditions has been compared for the recovery of the most commonly employed brominated flame retardants (BFRs) from styrenic polymeric matrixes. A HPLC-MS/MS method has been proposed for the simultaneous separation and quantification of tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) diastereomers, and decabromodiphenyl ether (deca-BDE) in the polymeric extracts. PLE results in complete extraction of TBBPA and HBCD (95-100% recovery), and intermediate recovery rates for deca-BDE (50%). MAE, on the other hand, gives comparable performance to PLE for HBCD, but lower extraction yields for TBBPA and mainly deca-BDE. Ultrasonication, finally, offers relatively low extraction recoveries (10-50%). The proposed analytical procedures could be used for the effective identification and quantification of BFRs in styrenic plastics and for quality purposes in recycling facilities that deal with styrenic fractions from waste electrical and electronic equipment (WEEE). PMID- 18511059 TI - Complex profiles of hydrophobic paralytic shellfish poisoning compounds in Gymnodinium catenatum identified by liquid chromatography with fluorescence detection and mass spectrometry. AB - The presence of hydrophobic analogues of paralytic shellfish poisoning toxins (PSTs) was studied in a Portuguese strain of Gymnodinium catenatum by conventional pre-column oxidation HPLC after a prolonged acetonitrile gradient coupled with fluorescence detection. Prior separation of hydrophobic PSTs analogues from hydrophilic analogues was done by solid-phase extraction (SPE) partitioning on a C18 cartridge. Several unknown oxidation products, with emission spectra similar to known PSTs, appeared after periodate or hydrogen peroxide oxidation. The compounds producing these oxidation products could be grouped into three major sub-groups according to SPE partitioning. The first one eluting with 10 and 20% MeOH, produced the first set of oxidation products observed after the saxitoxin oxidation product. The second one eluting with 30 100% MeOH produced the second set of oxidation products. The third one eluted with acidified 90% MeOH produced the third and last set of oxidation products. Additionally, the oxidation products corresponding to decarbamoyl gonyautoxins and decarbamoyl saxitoxins were also abundant, resulting from ester cleavage of the benzoate side chain of these compounds during the oxidation. Analysis of these fractions by LC-MS demonstrated the second sub-group was constituted by analogues of the 11-hydroxysulfated GC1/GC2, while the third sub-group was constituted by analogues of GC3, which lack the 11-hydroxysulfate. In addition to GC1/GC2 and GC3, novel analogues differing by 16u could be related, respectively, to the N1-hydroxyl analogues of GC1-GC3, designated GC4-GC6. A novel family of GC analogues, differing, by 16u from GC1-GC6, were hypothesized to possess an extra hydroxyl in the benzoate side chain, existing in both N1-hydroxylated and non-N1 hydroxylated variants, and tentatively designated GC1a-GC6a. The first sub-group was hypothesized to constitute an additional novel family of GC analogues with a hydroxysulfate group instead of the hydroxyl group in the benzoate side chain, tentatively designated GC1b-GC6b. PMID- 18511060 TI - Interstratification of trioctahedral and dioctahedral smectites through delamination and costacking. AB - Trioctahedral and dioctahedral organosmectites delaminate in 1-octanol to give stable monolayer colloidal dispersions. Addition of acetone to a mixture of these colloidal dispersions yields a composite of the two clays. Layers of the two smectites are interstratified in the composite. Due to random costacking of layers the thermal decomposition behavior of the composite is different from that of the parent smectites and their physical mixture. PMID- 18511061 TI - Magnesium hydroxide nanoparticles synthesized in water-in-oil microemulsions. AB - Well-dispersed magnesium hydroxide nanoplatelets were synthesized by a simple water-in-oil (w/o) microemulsion process, blowing gaseous ammonia (NH(3)) into microemulsion zones solubilized by magnesium chloride solution (MgCl(2)). Typical quaternary microemulsions of Triton X-100/cyclohexane/n-hexanol/water were used as space-confining microreactors for the nucleation, growth, and crystallization of magnesium hydroxide nanoparticles. The obtained magnesium hydroxide was characterized by field-emission scanning electron microscopy (FESEM), high resolution transmission election microscopy (HRTEM), X-ray powder diffraction (XRD), laser light scattering, Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis-differential scanning calorimetry (TGA-DSC). The mole ratio of water to surfactant (omega(0)) played an important role in the sizes of micelles and nanoparticles, increasing with the increase of omega(0). The compatibility and dispersibility of nanoparticles obtained from reverse micelles were improved in the organic phase. PMID- 18511062 TI - One-step synthesis of silver nanoparticles by sonication or heating using amphiphilic block copolymer as templates. AB - Block copolymer-supported Ag Nps (nanoparticles) have either a "cherry"-like or "raspberry"-like morphology [Antonietti, et al., Adv. Mater. 7 (1995) 1000-1005] depending on the amount of silver nitrate loading and the external conditions. Sonication favors silver nitrate and polyethyleneimine diffusion; the nucleation sites are well distributed in the micellar cores, so it is easy to form the cherry-like Ag NP colloids. However, when the amount of silver nitrate is decreased, it is heating that induces the formation of raspberry-like Ag NP colloids. The Ag NP colloids were investigated by transmission electron microscopy to demonstrate the nanosize dimensions and the location of the Ag NPs in the micelles. X-ray diffraction was employed to determine the crystal structure of the Ag NPs. UV-vis spectroscopy was employed for further qualitative characterization of the optical properties of Ag NPs. PMID- 18511063 TI - Validating parameters of a luciferase reporter gene assay to measure neutralizing antibodies to IFNbeta in multiple sclerosis patients. AB - Neutralizing antibodies (NAbs) can occur in some multiple sclerosis (MS) patients receiving interferon beta (IFNbeta) therapy. NAbs reduce drug bioavailabity and high NAb titers reduce drug efficacy. We describe the validation of the R. Farrell and G. Giovannoni luciferase reporter gene assay to measure NAbs to INFbeta. We assayed 163 sera from IFNbeta treated MS patients with an optimized luciferase method and compared the results to those obtained with the reference cytopathic effect (CPE) method using A549 cells and an encephalomyocarditis virus (EMCV). Binding antibodies (BAbs) were measured using a capture ELISA as a screening test for NAbs in the CPE assay. NAb status measured by the luciferase and the ELISA/CPE method did not yield a significant difference. Log10 NAb titers obtained from the luciferase assay and the A549/EMCV CPE methods correlated very well. The inter-assay coefficient of variation for titers was between 17.8-29.3%, and the intra-assay coefficient of variation was between 6.3-15.2%. The luciferase assay is reliable, appropriately sensitive and requires less time than the currently available NAb methods. PMID- 18511064 TI - Downregulation of ultraspiracle gene expression delays pupal development in honeybees. AB - Ecdysteroids regulate many aspects of insect physiology after binding to a heterodimer composed of the nuclear hormone receptor proteins ecdysone receptor (EcR) and ultraspiracle (Usp). Several lines of evidence have suggested that the latter also plays important roles in mediating the action of juvenile hormone (JH) and, thus, integrates signaling by the two morphogenetic hormones. By using an RNAi approach, we show here that Usp participates in the mechanism that regulates the progression of pupal development in Apis mellifera, as indicated by the observed pupal developmental delay in usp knocked-down bees. Knock-down experiments also suggest that the expression of regulatory genes such as ftz transcription factor 1 (ftz-f1) and juvenile hormone esterase (jhe) depend on Usp. Vitellogenin (vg), the gene coding the main yolk protein in honeybees, does not seem to be under Usp regulation, thus suggesting that the previously observed induction of vg expression by JH during the last stages of pupal development is mediated by yet unknown transcription factor complexes. PMID- 18511065 TI - Maternal effects on progeny size, number and body color in the desert locust, Schistocerca gregaria: Density- and reproductive cycle-dependent variation. AB - The effects of rearing density and maternal age on the progeny size, number and coloration of the desert locust, Schistocerca gregaria, were investigated. Isolated-reared females deposited smaller, but more eggs than crowd-reared females. The former produced smaller and more eggs with age, whereas the latter showed a tendency to produce larger and fewer eggs over time. A similar tendency was also observed with virgin females, indicating that mating or the presence of males was not important. The first egg pod produced by each mated crowd-reared female contained significantly smaller and more eggs than did the subsequent egg pods. The former often produced many green hatchlings (0-100%) characteristic of solitarious forms, whereas the egg pods deposited after the first pod produced predominantly black hatchlings typical of gregarious forms. Adults were highly sensitive to a shift in rearing density and quickly modified the quality and quantity of their progeny depending on the density encountered. The number of eggs per pod was influenced not only by the mother's rearing density but also by rearing density of the grandmother. The present results demonstrated that the characteristics of progeny are influenced not only by the crowding conditions experienced by the mother and grandmother but also by the mother's reproductive cycle. PMID- 18511066 TI - Interplay of JH, 20E and biogenic amines under normal and stress conditions and its effect on reproduction. AB - Juvenile hormone (JH) and 20-hydroxyecdysone (20E) are well known to play a gonadotropic role in adult insects. In Drosophila the mechanism of reciprocal regulation of JH and 20E is shown to be responsible for their proper balance. Dopamine is a mediator in this JH and 20E interplay. A proper balance between JH and 20E is crucial for the normal progress of oogenesis. An imbalance of gonadotropins leads to reproductive defects: a rise in JH titre leads to oviposition arrest, a rise in 20E level, to the degradation of vitellogenic oocytes. Upon a change in the level of one of the gonadotropins, the balance is restored owing to the relative change in the titre of the other. PMID- 18511067 TI - Further research on the production, longevity and infectivity of the zoospores of Leptolegnia chapmanii Seymour (Oomycota: Peronosporomycetes). AB - The effect of temperature on the production, survival and infectivity of zoospores of an Argentinean isolate of Leptolegnia chapmanii was determined under laboratory conditions. Production of zoospores of L. chapmaniiin vitro and in vivo upon first and fourth instars larvae of the mosquito Aedes aegypti was studied at three different temperatures. Zoospores from infected larvae were infective to mosquito larvae for 51, 12, and 5 consecutive days when maintained at 25, 35, and 10 degrees C, respectively. Maximum zoospore production in infected fourth-instar larvae was 9.6+/-1.4x10(4) zoosp/larva at 48 h at 25 degrees C. The average number of zoospores produced by individual fourth-instar Ae. aegypti larvae infected with L. chapmanii was 3.57+/-0.46x10(5) zoospores during 6 consecutive days at 25 degrees C. Zoospore production in vitro was also affected by temperature with a maximum of zoospores (n=47,666/ml) produced at 25 degrees C. When zoospores produced in vitro were used as inoculum against Ae. aegypti larvae at 25 degrees C, larval mortality was recorded for 5 consecutive weeks. The encystment process for zoospores took 17-20 min; the germination of cysts (excystment) occurred 5 min after exposure in water to mosquito larvae. The minimal time of contact between zoospores and mosquito larvae to develop infection was two minutes. Infection took place by zoospore attachment onto and then penetration through the larval cuticle or by ingestion of cysts as was confirmed by histological studies. Temperature directly affected infectivity and production of zoospores in vivo and in vitro although L. chapmanii zoospores tolerate a wide range of temperatures. PMID- 18511068 TI - A molecular revision of the taxonomic status of mermithid parasites of black flies from Quebec (Canada). AB - The four currently recognized mermithid (Nematoda) species parasitizing black flies (Diptera: Simuliidae) from Northeast America were distinguished using discriminatory PCR primers aimed at COI and 18S rDNA. Isomermis wisconsinensis, Gastromermis viridis and Mesomermis camdenensis were easily differentiated using either genomic target, even for juvenile mermithids damaged beyond morphological recognition. However, specimens from Mesomermis flumenalis being identical in external morphology and producing a unique-sized PCR product were classified by sequence data into four clearly distinguished molecular variants. This quartet was made of two winter and two summer 'physiological variants', including one which also belonged to, but diverged early from the rest of the Mesomermis genus. Combining the multiplex PCR and sequencing approaches allowed for the characterization of a multiple parasitism which simultaneously implicated I. wisconsinensis and two M. flumenalis variants. With another instance where parasites were identified by morphology only, this is the first report of black fly parasitism by multiple mermithid species. A phylogenetic tree built by combining our sequences to previous GenBank entries likely indicates a monophyletic origin for the mermithid family, but also suggests that differentiation between parasite genera sometimes occurred before the evolutionary emergence of the actual host group. PMID- 18511069 TI - Engineered bacterial outer membrane vesicles with enhanced functionality. AB - We have engineered bacterial outer membrane vesicles (OMVs) with dramatically enhanced functionality by fusing several heterologous proteins to the vesicle associated toxin ClyA of Escherichia coli. Similar to native unfused ClyA, chimeric ClyA fusion proteins were found localized in bacterial OMVs and retained activity of the fusion partners, demonstrating for the first time that ClyA can be used to co-localize fully functional heterologous proteins directly in bacterial OMVs. For instance, fusions of ClyA to the enzymes beta-lactamase and organophosphorus hydrolase resulted in synthetic OMVs that were capable of hydrolyzing beta-lactam antibiotics and paraoxon, respectively. Similarly, expression of an anti-digoxin single-chain Fv antibody fragment fused to the C terminus of ClyA resulted in designer "immuno-MVs" that could bind tightly and specifically to the antibody's cognate antigen. Finally, OMVs displaying green fluorescent protein fused to the C terminus of ClyA were highly fluorescent and, as a result of this new functionality, could be easily tracked during vesicle interaction with human epithelial cells. We expect that the relative plasticity exhibited by ClyA as a fusion partner should prove useful for: (i) further mechanistic studies to identify the vesiculation machinery that regulates OMV secretion and to map the intracellular routing of ClyA-containing OMVs during invasion of host cells; and (ii) biotechnology applications such as surface display of proteins and delivery of biologics. PMID- 18511070 TI - Structural analysis of CYP2R1 in complex with vitamin D3. AB - The activation of vitamin D to its hormonal form is mediated by cytochrome P450 enzymes. CYP2R1 catalyzes the initial step converting vitamin D into 25 hydroxyvitamin D. A CYP2R1 gene mutation causes an inherited form of rickets due to 25-hydroxylase deficiency. To understand the narrow substrate specificity of CYP2R1 we obtained the hemeprotein in a highly purified state, confirmed the enzyme as a vitamin D 25-hydroxylase, and solved the crystal structure of CYP2R1 in complex with vitamin D3. The CYP2R1 structure adopts a closed conformation with the substrate access channel being covered by the ordered B'-helix and slightly opened to the surface, which defines the substrate entrance point. The active site is lined by conserved, mostly hydrophobic residues. Vitamin D3 is bound in an elongated conformation with the aliphatic side-chain pointing toward the heme. The structure reveals the secosteroid binding mode in an extended active site and allows rationalization of the molecular basis of the inherited rickets associated with CYP2R1. PMID- 18511071 TI - Pre-folding IkappaBalpha alters control of NF-kappaB signaling. AB - Transcription complex components frequently show coupled folding and binding but the functional significance of this mode of molecular recognition is unclear. IkappaBalpha binds to and inhibits the transcriptional activity of NF-kappaB via its ankyrin repeat (AR) domain. The beta-hairpins in ARs 5-6 in IkappaBalpha are weakly-folded in the free protein, and their folding is coupled to NF-kappaB binding. Here, we show that introduction of two stabilizing mutations in IkappaBalpha AR 6 causes ARs 5-6 to fold cooperatively to a conformation similar to that in NF-kappaB-bound IkappaBalpha. Free IkappaBalpha is degraded by a proteasome-dependent but ubiquitin-independent mechanism, and this process is slower for the pre-folded mutants both in vitro and in cells. Interestingly, the pre-folded mutants bind NF-kappaB more weakly, as shown by both surface plasmon resonance and isothermal titration calorimetry in vitro and immunoprecipitation experiments from cells. One consequence of the weaker binding is that resting cells containing these mutants show incomplete inhibition of NF-kappaB activation; they have significant amounts of nuclear NF-kappaB. Additionally, the weaker binding combined with the slower rate of degradation of the free protein results in reduced levels of nuclear NF-kappaB upon stimulation. These data demonstrate clearly that the coupled folding and binding of IkappaBalpha is critical for its precise control of NF-kappaB transcriptional activity. PMID- 18511072 TI - The anti-cancer drug chlorambucil as a substrate for the human polymorphic enzyme glutathione transferase P1-1: kinetic properties and crystallographic characterisation of allelic variants. AB - The commonly used anti-cancer drug chlorambucil is the primary treatment for patients with chronic lymphocytic leukaemia. Chlorambucil has been shown to be detoxified by human glutathione transferase Pi (GST P1-1), an enzyme that is often found over-expressed in cancer tissues. The allelic variants of GST P1-1 are associated with differing susceptibilities to leukaemia and differ markedly in their efficiency in catalysing glutathione (GSH) conjugation reactions. Here, we perform detailed kinetic studies of the allelic variants with the aid of three representative co-substrates. We show that the differing catalytic properties of the variants are highly substrate-dependent. We show also that all variants exhibit the same temperature stability in the range 10 degrees C to 45 degrees C. We have determined the crystal structures of GST P1-1 in complex with chlorambucil and its GSH conjugate for two of these allelic variants that have different residues at positions 104 and 113. Chlorambucil is found to bind in a non-productive mode to the substrate-binding site (H-site) in the absence of GSH. This result suggests that under certain stress conditions where GSH levels are low, GST P1-1 can inactivate the drug by sequestering it from the surrounding medium. However, in the presence of GSH, chlorambucil binds in the H-site in a productive mode and undergoes a conjugation reaction with GSH present in the crystal. The crystal structure of the GSH-chlorambucil complex bound to the *C variant is identical with the *A variant ruling out the hypothesis that primary structure differences between the variants cause structural changes at the active site. Finally, we show that chlorambucil is a very poor inhibitor of the enzyme in contrast to ethacrynic acid, which binds to the enzyme in a similar fashion but can act as both substrate and inhibitor. PMID- 18511073 TI - Identification of ligands affecting the activity of the transcriptional repressor CcpN from Bacillus subtilis. AB - Carbon catabolite repression in Bacillus subtilis is mediated primarily by the major regulator CcpA. However, sugar-dependent repression of three genes, sr1 encoding a small nontranslated RNA and two genes coding for gluconeogenic enzymes, gapB and pckA, is carried out by the transcriptional repressor CcpN (control catabolite protein of gluconeogenic genes). It has previously been shown that ccpN is constitutively expressed, which leads to a constant occupation of all operators with CcpN. Since this would not allow for specific regulation, a ligand that modulates CcpN activity is required. In vitro transcription assays demonstrated that CcpN is able to specifically repress transcription to a small extent at the three mentioned promoters in the absence of an activating ligand. Upon testing of several ligands, including nucleotides and glycolysis intermediates, it could be shown that ATP is able to specifically enhance the repressing activity of CcpN, and this effect was more pronounced at a slightly acidic pH. Furthermore, ADP was found to specifically counteract the repressive effect of ATP. Circular dichroism measurements demonstrated a significant alteration of CcpN structure in the presence of ATP at acidic pH and in the presence of ADP. Electrophoretic mobility shift assays revealed that neither ATP nor ADP altered the affinity of CcpN for its operators. Therefore, we hypothesise that the effect of ligand-bound CcpN on the RNA polymerase might be due to a conformational switch that alters the interaction between the two proteins. Based on these results, a working model for CcpN action is discussed. PMID- 18511074 TI - Coils in the membrane core are conserved and functionally important. AB - With the increasing number of available alpha-helical transmembrane (TM) protein structures, the traditional picture of membrane proteins has been challenged. For example, reentrant regions, which enter and exit the membrane at the same side, and interface helices, which lie parallel with the membrane in the membrane-water interface, are common. Furthermore, TM helices are frequently kinked, and their length and tilt angle vary. Here, we systematically analyze 7% of all residues within the deep membrane core that are in coil state. These coils can be found in TM-helix kinks as major breaks in TM helices and as parts of reentrant regions. Coil residues are significantly more conserved than other residues. Due to the polar character of the coil backbone, they are either buried or located near aqueous channels. Coil residues are frequently found within channels and transporters, where they introduce the flexibility and polarity required for transport across the membrane. Therefore, we believe that coil residues in the membrane core, while constituting a structural anomaly, are essential for the function of proteins. PMID- 18511075 TI - Structural impact of the E113Q counterion mutation on the activation and deactivation pathways of the G protein-coupled receptor rhodopsin. AB - Disruption of an interhelical salt bridge between the retinal protonated Schiff base linked to H7 and Glu113 on H3 is one of the decisive steps during activation of rhodopsin. Using previously established stabilization strategies, we engineered a stabilized E113Q counterion mutant that converted rhodopsin to a UV absorbing photoreceptor with deprotonated Schiff base and allowed reconstitution into native-like lipid membranes. Fourier-transform infrared difference spectroscopy reveals a deprotonated Schiff base in the photoproducts of the mutant up to the active state Meta II, the absence of the classical pH-dependent Meta I/Meta II conformational equilibrium in favor of Meta II, and an anticipation of active state features under conditions that stabilize inactive photoproduct states in wildtype rhodopsin. Glu181 on extracellular loop 2, is found to be unable to maintain a counterion function to the Schiff base on the activation pathway of rhodopsin in the absence of the primary counterion, Glu113. The Schiff base becomes protonated in the transition to Meta III. This protonation is, however, not associated with a deactivation of the receptor, in contrast to wildtype rhodopsin. Glu181 is suggested to be the counterion in the Meta III state of the mutant and appears to be capable of stabilizing a protonated Schiff base in Meta III, but not of constraining the receptor in an inactive conformation. PMID- 18511076 TI - Changes in BDNF serum levels in patients with major depression disorder (MDD) after 6 months treatment with sertraline, escitalopram, or venlafaxine. AB - Recent studies have implicated brain-derived neurotrophic factor (BDNF) in the pathophysiology of depression and the activity of antidepressant drugs. Serum BDNF levels are lower in depressed patients, and increase in response to antidepressant medication. However, how BDNF responds to different classes of antidepressant drugs is unknown. We assessed serum BDNF levels in 21 patients with major depressive episode treated with sertraline, escitalopram, or venlafaxine and 20 healthy controls. Serum samples were collected between 10 a.m. and 12 p.m. at baseline, 5 weeks, and 6 months of treatment. BDNF levels were measured via immunoassay. The severity of symptoms and response to treatment were assessed by the Hamilton rating scales for depression (HRSD). Baseline serum BDNF levels were significantly lower in depressed patients compared to controls. Sertraline increased BDNF levels after 5 weeks and 6 months of treatment. Venlafaxine increased BDNF levels only after 6 months. Escitalopram did not affect BDNF levels at either time point. A significant negative association was found between percentage increase in BDNF levels and percentage decreased in HRSD scores after 6 months of treatment. In conclusion, these results suggest that different antidepressant drugs have variable effects on serum BDNF levels. This is true even though the three different drugs were equally effective in relieving symptoms of depression and anxiety. PMID- 18511077 TI - Bimodal response: electric tissue ablation--long term studies of morbidity and pathological change. PMID- 18511078 TI - Application of a new type of sutureless magnetic biliary-enteric anastomosis stent for one-stage reconstruction of the biliary-enteric continuity after acute bile duct injury: an experimental study. AB - BACKGROUND: The bile duct cannot be repaired or reconstructed in one stage after 24 h of bile duct injury due to significant inflammation. Even if the bile duct can be repaired, anastomosis is difficult because of the extremely technical nature of the procedure. The traditional method of anastomosis utilizes screw thread. This would increase the inflammatory response, delay anastomotic healing, and lead to the increase in the failure rate. To reconstruct the biliary-enteric continuity under the circumstance of severe inflammation after bile duct injury, we invented a new type of anastomotic apparatus (sutureless magnetic stent) for cholangiojejunostomy. The objective of this study was to evaluate the effect of a new type of sutureless magnetic biliary-enteric anastomosis stent, which was used to reconstruct the biliary-enteric continuity in one stage. The reconstruction was conducted under the circumstance of severe inflammation after acute bile duct injury in dogs. METHODS: We used a model of acute bile duct injury and bile peritonitis in dogs. The sutureless magnetic biliary-enteric anastomosis stents was used to reconstruct the biliary-enteric continuity in one stage under the circumstance of a bile duct with severe inflammation. The effect of stents was observed. Cholangiography and anastomotic histology were examined at 1 mo and compared with traditional manual anastomosis. RESULTS: Anastomotic stents were used to reconstruct the biliary-enteric continuity in one stage in dogs. No anastomotic leak or infection occurred. Cholangiography showed that the anastomosis was unobstructed. Histological examinations showed that the anastomosis healed well, the inflammatory reaction was small, and collagen fibers lined up in order. There was high incidence of bile leakage in the conventional suture group. Cholangiography showed that anastomotic stenosis was high. Histological examination showed that there was more extensive inflammation around the anastomosis and the collagen fibers were disorganized. CONCLUSION: It was safe and feasible to use the new type of anastomosis stent to reconstruct the biliary-enteric continuity in one stage under the circumstance of severe bile duct inflammation after bile duct injury in dogs. PMID- 18511079 TI - A report card system using error profile analysis and concurrent morbidity and mortality review: surgical outcome analysis, part II. AB - BACKGROUND: An effective report card system for adverse outcome error analysis following surgery is lacking. We hypothesized that a memorialized database could be used in conjunction with error analysis and management evaluation at Morbidity & Mortality conference to generate individualized report cards for Attending Surgeon and System performance. STUDY DESIGN: Prospectively collected data from September 2000 through April 2005 were reported following Morbidity & Mortality review on 1618 adverse outcomes, including 219 deaths, following 29,237 operative procedures, in a complete loop to approximately 60 individual surgeons and responsible system personnel. RESULTS: A 40% reduction of gross mortality (P < 0.001) and 43% reduction of age-adjusted mortality were achieved over 4 years at the Academic Center. Quality issues were identified at a rate three times greater than required by New York State regulations and increased from a baseline 4.96% to 32.7% (odds ratio 1.94; P < 0.03) in cases associated with mortality. A detailed review demonstrated a significant increase (P < 0.001) in system errors and physician-related diagnostic and judgment errors associated with mortality highlighted those practices and processes involved, and contrasted the results between academic (43% mortality improvement) and community (no improvement) hospitals. CONCLUSIONS: The findings suggest that structured concurrent data collection combined with non-punitive error-based case review and individualized report cards can be used to provide detailed feedback on surgical performance to individual surgeons and possibly improve clinical outcomes. PMID- 18511080 TI - Estradiol-treated mesenchymal stem cells improve myocardial recovery after ischemia. AB - BACKGROUND: Stem cell therapy is a promising treatment modality for injured cardiac tissue. A novel mechanism for this cardioprotection may include paracrine actions. Our lab has recently shown that gender differences exist in mesenchymal stem cell (MSC) paracrine function. Estrogen is implicated in the cardioprotection found in females. It remains unknown whether 17beta-estradiol (E2) affects MSC paracrine function and whether E2-treated MSCs may better protect injured cardiac tissue. We hypothesize that E2-exposed MSCs infused into hearts prior to ischemia may demonstrate increased vascular endothelial growth factor (VEGF) production and greater protection of myocardial function compared to untreated MSCs. MATERIALS AND METHODS: Untreated and E2-treated MSCs were isolated, cultured, and plated and supernatants were harvested for VEGF assay (enzyme-linked immunosorbent assay). Adult male Sprague-Dawley rat hearts (n = 13) were isolated and perfused via Langendorff model and subjected to 15 min equilibration, 25 min warm global ischemia, and 40 min reperfusion. Hearts were randomly assigned to perfusate vehicle, untreated male MSC, or E2-treated male MSC. Transcoronary delivery of 1 million MSCs was performed immediately prior to ischemia in experimental hearts. RESULTS: E2-treated MSCs provoked significantly more VEGF production than untreated MSCs (933.2 +/- 64.9 versus 595.8 +/- 10.7 pg/mL). Postischemic recovery of left ventricular developed pressure was significantly greater in hearts infused with E2-treated MSCs (66.9 +/- 3.3%) than untreated MSCs (48.7 +/- 3.7%) and vehicle (28.9 +/- 4.6%) at end reperfusion. There was also greater recovery of the end diastolic pressure with E2-treated MSCs than untreated MSCs and vehicle. CONCLUSIONS: Preischemic infusion of MSCs protects myocardial function and viability. E2-treated MSCs may enhance this paracrine protection, which suggests that ex vivo modification of MSCs may improve therapeutic outcome. PMID- 18511081 TI - Statistics of canonical RNA pseudoknot structures. AB - In this paper we study canonical RNA pseudoknot structures. We prove central limit theorems for the distributions of the arc-numbers of k-noncrossing RNA structures with given minimum stack-size tau over n nucleotides. Furthermore we compare the space of all canonical structures with canonical minimum free energy pseudoknot structures. Our results generalize the analysis of Schuster et al. obtained for RNA secondary structures [Hofacker, I.L., Schuster, P., Stadler, P.F., 1998. Combinatorics of RNA secondary structures. Discrete Appl. Math. 88, 207-237; Jin, E.Y., Reidys, C.M., 2007b. Central and local limit theorems for RNA structures. J. Theor. Biol. 250 (2008), 547-559; 2007a. Asymptotic enumeration of RNA structures with pseudoknots. Bull. Math. Biol., 70 (4), 951-970] to k noncrossing RNA structures. Here k2 and tau are arbitrary natural numbers. We compare canonical pseudoknot structures to arbitrary structures and show that canonical pseudoknot structures exhibit significantly smaller exponential growth rates. We then compute the asymptotic distribution of their arc-numbers. Finally, we analyze how the minimum stack-size and crossing number factor into the distributions. PMID- 18511082 TI - A network model for activity-dependent sleep regulation. AB - We develop and characterize a dynamical network model for activity-dependent sleep regulation. Specifically, in accordance with the activity-dependent theory for sleep, we view organism sleep as emerging from the local sleep states of functional units known as cortical columns; these local sleep states evolve through integration of local activity inputs, loose couplings with neighboring cortical columns, and global regulation (e.g. by the circadian clock). We model these cortical columns as coupled or networked activity-integrators that transition between sleep and waking states based on thresholds on the total activity. The model dynamics for three canonical experiments (which we have studied both through simulation and system-theoretic analysis) match with experimentally observed characteristics of the cortical-column network. Most notably, assuming connectedness of the network graph, our model predicts the recovery of the columns to a synchronized state upon temporary overstimulation of a single column and/or randomization of the initial sleep and activity integration states. In analogy with other models for networked oscillators, our model also predicts the possibility for such phenomena as mode-locking. PMID- 18511083 TI - Prediction of C-to-U RNA editing sites in plant mitochondria using both biochemical and evolutionary information. AB - Although cytidine-to-uridine conversions in plant mitochondria were discovered 18 years ago, it was still an enigmatic process. Since the sequencing projects of plant mitochondrial genomes are providing more and more available sequences, the requirements of computationally identifying C-to-U RNA editing sites are also increasing. By incorporating both evolutionary and biochemical information, we developed a novel algorithm for predicting C-to-U RNA editing sites in plant mitochondria. The algorithm has been implemented as an online service called CURE (Cytidine-to-Uridine Recognizing Editor). CURE performs better than other methods that are based on only biochemical or only evolutionary information. CURE also provides the ability of predicting C-to-U RNA editing sites in non-coding regions and the synonymous C-to-U RNA editing sites in coding regions that are impossible for other methods. Furthermore, CURE can carry out prediction directly on the entire mitochondria genome sequence. The prediction results of CURE suggest the functional importance of synonymous RNA editing sites, which was neglected before. The CURE service can be accessed at http://bioinfo.au.tsinghua.edu.cn/cure. PMID- 18511084 TI - Female alternative reproductive behaviors: the effect of female group size on mate assessment and copying. AB - Extensive theoretical and empirical research has focused on male alternative reproductive tactics. In comparison, female alternative tactics have attracted little attention, and further theoretical and empirical research are needed. Using a game theoretical model, we examine female choice alternatives (1) by considering assessment errors in a novel and more realistic manner than done previously, and (2) for the first time, by highlighting the formation of groups of females as an important consequence of copying behavior. We consider two alternatives: direct assessment of male quality by females and female copying of the choice of other females. Assessment and copying are predicted to coexist under a wide variety of circumstances and copying is favored when females make assessment errors, when high-quality males are either common or very rare, and when female fitness declines with the number of other females choosing the same male. We also find that the frequency of copying at equilibrium is predicted to decrease when the presence of other females mating with the same male has a positive effect on female fitness (e.g. through increased male parental effort, decreased predation risk or cooperation among females). Female alternative choice tactics also influence the potential for sexual selection. In our model, when the frequency of copying females is low, the potential for sexual selection can be higher than in the absence of female copying. However, contrary to previous theory, we find that as copying females become more common than assessing females, the potential for sexual selection will be low as more females copy the mate choice of other copiers without assessment. PMID- 18511085 TI - The nervous system might 'orthogonalize' to discriminate. AB - It is still unclear how information is actually stored in biological neural networks. We propose here that information could be first orthogonalized and then stored. This could happen in a manner similar to how a set of vectors is transformed into a set of orthogonalized (i.e. mutually perpendicular) vectors. Orthogonalization may overcome the limits of conventional artificial networks, particularly the catastrophic interference caused by interference between stored inputs. The features needed to allow orthogonalization are common to biological networks, suggesting that it may be a common network mechanism. To illustrate this hypothesis, we characterize the underlying features that an archetypal biological network must have in order to perform orthogonalization, and point out that a number of actual networks show this archetypal network organization. PMID- 18511086 TI - Spatial heterogeneity of mortality and temporal fluctuation in fertility promote coexistence but not vice versa: a random-community approach. AB - Both spatial heterogeneity and temporal fluctuation of the environment are important mechanisms promoting species coexistence, but they work in different manners. We consider many pairs of species with randomly generated survivorship and fertility in the lottery model, and examine how the variability in demographic processes affects the outcome of competition. The results are: [corrected] (1) Coexistence is easier if habitat difference in mortality is greater, or if year-to-year variation in reproductive rate is larger. But neither habitat [corrected] difference in fertility nor temporal variation in mortality promotes coexistence. [corrected] (2) Mean fertility does not affect the outcome if the coefficient of variation [corrected] remains constant. In contrast, enhanced mean mortality decreases the fraction of coexisting pairs if the environment fluctuates temporally. [corrected] (3) We also investigate the effect of limited dispersal of propagules between habitats. Compared with the complete mixing case, the fraction of coexisting pairs is clearly enhanced if the spatial heterogeneity is the major source of environmental variation, but shows slight increase if the temporal fluctuation is dominant. We conclude that spatial heterogeneity is likely to work more effectively in promoting species coexistence than temporal fluctuation, especially when the species suffer relatively high mortality, and disperse their propagules in a limited spatial scale. PMID- 18511087 TI - Benthic input rates predict seagrass (Posidonia oceanica) fish farm-induced decline. AB - Fish farms represent a growing source of anthropogenic disturbance to benthic communities, and efficient predictors of such impacts are urgently needed. We explored the effects of fish farm benthic organic and nutrient inputs on the population dynamics of a key seagrass species (Posidonia oceanica) in four Mediterranean deep meadows adjacent to sea bream and sea bass farms. We performed two annual plant censuses on permanent plots at increasing distance from farms and measured benthic sedimentation rates around plots. High shoot mortality rates were recorded near the cages, up to 20 times greater than at control sites. Recruitment rates increased in variability but could not compensate mortality, leading to rapid seagrass decline within the first 100 m from cages. Seagrass mortality increased with total sedimentation rates (K=0.55, p<0.0002), and with organic matter (K=0.50, p=0.001), total nitrogen (K=0.46, p=0.002) and total phosphorus (K=0.56, p<3.10(-5)) inputs. P. oceanica decline accelerated above a phosphorus loading threshold of 50mg m(-2)day(-1). Phosphorus benthic sedimentation rate seems a powerful predictor of seagrass mortality from fish farming. Coupling direct measurements of benthic sedimentation rates with dynamics of key benthic species is proposed as an efficient strategy to predict fish farm impacts to benthic communities. PMID- 18511088 TI - A role for nuclear beta-catenin in SNRI antidepressant-induced hippocampal cell proliferation. AB - Increasing evidences have been accumulated during recent years suggesting a role for antidepressant drugs (ADs) as hippocampal neurogenesis enhancers, but the information about the transductional mechanisms involved in this response is very limited. We have studied in the adult rat hippocampus the effects of chronic treatment with the dual reuptake inhibitor (SNRI) venlafaxine on both cellular proliferation rate and expression of key effectors of several signaling pathways. Increased cell proliferation (BrdU incorporation) in subgranular zone (SGZ) was achieved after chronic treatment with a high dose (40 mg/kg/day) of venlafaxine. However, significant increases in the immunoreactivity of hippocampal beta catenin in SGZ were already detected after administration of a lower dose of the drug (10 mg/kg/day). Western blot and immunoelectron microscopy studies demonstrated an increased presence of beta-catenin at the nuclear level. An increase in cytosolic AKT levels was also observed in venlafaxine-treated animals. These results suggest that the hippocampal proliferative effect of chronic venlafaxine, only evident when both serotonin (5-HT) and noradrenaline/norepinephrine (NE) reuptake systems are inhibited, requires a strong activation of intracellular signaling through Wnt (beta-catenin translocation) and AKT/PKB pathways. This activation would probably result in an increase of the expression of cell cycle regulator genes. Furthermore pERK2/ERK2 rate was also increased in the hippocampus of AD-treated animals, while no differences in the levels of CREB and p-CREB were observed. These results illustrate the complexity of the intracellular events underlying the neurogenetic responses of ADs. They also support the relevance of such effects for the therapeutic effects of these drugs. PMID- 18511089 TI - Global retrograde amnesia but selective anterograde amnesia after frontal temporal disconnection in monkeys. AB - Prefrontal cortex and inferior temporal cortex interact in support of a wide variety of learning and memory functions. In macaque monkeys, a disconnection of prefrontal and temporal cortex produces severe new learning impairments in a range of complex learning tasks such as visuo-motor conditional learning and object-in-place scene learning. The retrograde effects of this disconnection, however, have never been fully examined. We therefore assessed the postoperative retention of 128 preoperatively learned object discrimination problems in monkeys with prefrontal-temporal disconnection using 1 trial postoperative retention tests. Because previous experiments have suggested that both spatial and temporal factors may be important in engaging frontal-temporal interaction we used object discrimination problems with a variety of spatial and temporal properties. Postoperatively, although monkeys with prefrontal-temporal disconnection displayed a retrograde amnesia for all problem types, subsequent assessments of new learning revealed selective anterograde amnesia, which was limited to problems in which objects were presented as serial compound stimuli. The pattern of broad retrograde amnesia with selective anterograde amnesia contrasts with recent data from monkeys with lesions which disrupt subcortical-cortical connectivity and which show the opposite pattern, namely no retrograde amnesia but severe anterograde amnesia. These results support the hypothesis that visual memory acquisition is supported by subcortical-cortical interactions while the retrieval of visual memories normally depends on the interaction between prefrontal cortex and inferior temporal cortex. PMID- 18511090 TI - Three types of sesquiterpenes from rhizomes of Atractylodes lancea. AB - Eight sesquiterpenes, including four guaiane-types containing an interesting epoxy unit (1-4), a rare tricyclic carbon skeleton-type (5) and three eudesmane types (6-8), along with five known compounds, were isolated from rhizomes of Atractylodes lancea. The structures and relative configurations of 1-8 were determined by analysis of spectroscopic data, and the absolute configuration of 8 was assigned by application of the CD technique. Compounds 1, 2 and 4 were evaluated for their cytotoxic effects against P388 and A549 cells, but all were inactive. Possible biosynthetic pathways for sesquiterpenes (1-8) were discussed. PMID- 18511091 TI - Proceedings of the Fifth International Meeting on Rapid Responses to Steroid Hormones. PMID- 18511092 TI - In vitro and in vivo percutaneous absorption of retinol from cosmetic formulations: significance of the skin reservoir and prediction of systemic absorption. AB - The percutaneous absorption of retinol (Vitamin A) from cosmetic formulations was studied to predict systemic absorption and to understand the significance of the skin reservoir in in vitro absorption studies. Viable skin from fuzzy rat or human subjects was assembled in flow-through diffusion cells for in vitro absorption studies. In vivo absorption studies using fuzzy rats were performed in glass metabolism cages for collection of urine, feces, and body content. Retinol (0.3%) formulations (hydroalcoholic gel and oil-in-water emulsion) containing (3)H-retinol were applied and absorption was measured at 24 or 72 h. All percentages reported are % of applied dose. In vitro studies using human skin and the gel and emulsion vehicles found 0.3 and 1.3% retinol, respectively, in receptor fluid at 24 h. Levels of absorption in the receptor fluid increased over 72 h with the gel and emulsion vehicles. Using the gel vehicle, in vitro rat skin studies found 23% in skin and 6% in receptor fluid at 24 h, while 72-h studies found 18% in skin and 13% in receptor fluid. Thus, significant amounts of retinol remained in rat skin at 24 h and decreased over 72 h, with proportional increases in receptor fluid. In vivo rat studies with the gel found 4% systemic absorption of retinol after 24 h and systemic absorption did not increase at 72 h. Retinol remaining in rat skin after in vivo application was 18% and 13% of the applied dermal dose after 24 and 72 h, respectively. Similar observations were made with the oil-in water emulsion vehicle in the rat. Retinol formed a reservoir in rat skin both in vivo and in vitro. Little additional retinol was bioavailable after 24 h. Comparison of these in vitro and in vivo results for absorption through rat skin indicates that the 24-h in vitro receptor fluid value accurately estimated 24-h in vivo systemic absorption. Therefore, the best single estimate of retinol systemic absorption from in vitro human skin studies is the 24-h receptor fluid value. However, the receptor fluid value from the 72-h extended study may be used in a worst-case exposure estimate. In conclusion, in vivo skin absorption studies can be useful in determining whether to include material in the in vitro skin reservoir as absorbable material in estimates of systemic absorption. PMID- 18511093 TI - Activation of AMP-activated protein kinase by tributyltin induces neuronal cell death. AB - AMP-activated protein kinase (AMPK), a member of the metabolite-sensing protein kinase family, is activated by energy deficiency and is abundantly expressed in neurons. The environmental pollutant, tributyltin chloride (TBT), is a neurotoxin, and has been reported to decrease cellular ATP in some types of cells. Therefore, we investigated whether TBT activates AMPK, and whether its activation contributes to neuronal cell death, using primary cultures of cortical neurons. Cellular ATP levels were decreased 0.5 h after exposure to 500 nM TBT, and the reduction was time-dependent. It was confirmed that most neurons in our culture system express AMPK, and that TBT induced phosphorylation of AMPK. Compound C, an AMPK inhibitor, reduced the neurotoxicity of TBT, suggesting that AMPK is involved in TBT-induced cell death. Next, the downstream target of AMPK activation was investigated. Nitric oxide synthase, p38 phosphorylation and Akt dephosphorylation were not downstream of TBT-induced AMPK activation because these factors were not affected by compound C, but glutamate release was suggested to be controlled by AMPK. Our results suggest that activation of AMPK by TBT causes neuronal death through mediating glutamate release. PMID- 18511094 TI - Ultrasound enhanced thrombolysis in acute arterial ischemia. AB - In vitro and animal studies have shown that thrombolysis with intravenous tissue plasminogen activator (tPA) can be enhanced with ultrasound. Ultrasound delivers mechanical pressure waves to the clot, thus exposing more thrombus surface to circulating drug. Moreover, intravenous gaseous microspheres with ultrasound have been shown to be a potential alternative to fibrinolytic agents to recanalize discrete peripheral thrombotic arterial occlusions or acute arteriovenous graft thromboses. Small phase I-II randomized and non-randomized clinical trials have shown promising results concerning the potential applications of ultrasound enhanced thrombolysis in the setting of acute cerebral ischemia. CLOTBUST was an international four-center phase II trial, which demonstrated that, in patients with acute ischemic stroke, transcranial Doppler (TCD) monitoring augments tPA induced arterial recanalization (sustained complete recanalization rates: 38% vs. 13%) with a non-significant trend toward an increased rate of clinical recovery from stroke, as compared with placebo. The rates of symptomatic intracerebral hemorrhage (sICH) were similar in the active and placebo group (4.8% vs. 4.8%). Smaller single-center clinical trials using transcranial color-coded sonography (TCCD) reported recanalization rates ranging from 27% to 64% and sICH rates of 0 18%. A separate clinical trial evaluating the safety and efficacy of therapeutic low-frequency ultrasound was discontinued because of a concerning sICH rate of 36% in the active group. To further enhance the ability of tPA to break up thrombi, current ongoing clinical trials include phase II studies of a single beam 2 MHz TCD with perflutren-lipid microspheres. Moreover, potential enhancement of intra-arterial tPA delivery is being clinically tested with 1.7 2.1 MHz pulsed wave ultrasound (EKOS catheter) in ongoing phase II-III clinical trials. Intravenous platelet-targeted microbubbles with low-frequency ultrasound are currently investigated as a rapid noninvasive technique to identify thrombosed intracranial and peripheral vessels. Multi-national dose escalation studies of microspheres and the development of an operator independent ultrasound device are underway. PMID- 18511096 TI - Troxler effect with dichoptic stimulus presentations: evidence for binocular inhibitory summation and interocular suppression. AB - Whether the Troxler effect (TE) has to do with interocular suppression and/or summation was studied with dichoptically matched (binocular or dioptic) and unmatched (monocular) stimulus presentations. Perceptual disappearance was found to occur more slowly under the binocular condition (mean=14.2s) than the monocular condition (mean=8.4s), but much faster than predicted by probability summation of the experimentally obtained latencies and durations of the TE in the monocular conditions (>27 s), suggesting a binocular inhibitory summation, the opposite of the binocular summation found with detection and contrast matching tasks [(Blake, R., & Fox, R. (1973). The psychological inquiry into binocular summation. Perception and Psychophysics, 14, 161-185; Blake, R., Sloane, M., & Fox, R. (1981). Further developments in binocular summation. Perception &Psychophysics 30, 266-276.)]. In addition, Ss with poorer stereoacuity took longer to see the disappearance in the monocular condition, and showed a larger disparity between the TEs from the two monocular conditions, suggesting a contribution of interocular suppression to the TE. PMID- 18511097 TI - Functional hierarchies of nonconscious visual processing. AB - A number of psychophysical techniques can be used to eliminate the registration of stimuli in visual awareness and to study the dynamics of conscious and nonconscious information processing in the visual system. However, little is known about how these techniques relate to each other. We chose to compare binocular rivalry, induced by orthogonal gratings presented separately to the two eyes, and metacontrast suppression, produced when a target stimulus is followed by a spatially surrounding mask stimulus, to investigate relative levels and correlates of nonconscious processing. Combined with prior results, our findings indicate that binocular rivalry expresses its suppressive effects prior to the level at which the mechanism of metacontrast does. Implications for theories of masking and interpretations of the loss or perceptual effects when stimulus visibility is suppressed by different psychophysical methods are discussed. PMID- 18511098 TI - The organization of spatial frequency maps measured by cortical flavoprotein autofluorescence. AB - To determine the organization of spatial frequency (SF) preference within cat Area 17, we imaged responses to stimuli with different SFs using optical intrinsic signals (ISI) and flavoprotein autofluorescence (AFI). Previous studies have suggested that neurons cluster based on SF preference, but a recent report argued that SF maps measured with ISI were artifacts of the vascular bed. Because AFI derives from a non-hemodynamic signal, it is less contaminated by vasculature. The two independent imaging methods produced similar SF preference maps in the same animals, suggesting that the patchy organization of SF preference is a genuine feature of Area 17. PMID- 18511095 TI - Progress and problems in the application of focused ultrasound for blood-brain barrier disruption. AB - Advances in neuroscience have resulted in the development of new diagnostic and therapeutic agents for potential use in the central nervous system (CNS). However, the ability to deliver the majority of these agents to the brain is limited by the blood-brain barrier (BBB), a specialized structure of the blood vessel wall that hampers transport and diffusion from the blood to the brain. Many CNS disorders could be treated with drugs, enzymes, genes, or large-molecule biotechnological products such as recombinant proteins, if they could cross the BBB. This article reviews the problems of the BBB presence in treating the vast majority of CNS diseases and the efforts to circumvent the BBB through the design of new drugs and the development of more sophisticated delivery methods. Recent advances in the development of noninvasive, targeted drug delivery by MRI-guided ultrasound-induced BBB disruption are also summarized. PMID- 18511099 TI - Removal of perfluorinated surfactants by sorption onto granular activated carbon, zeolite and sludge. AB - Perfluorinated surfactants are emerging pollutants of increasing public health and environmental concern due to recent reports of their world-wide distribution, environmental persistence and bioaccumulation potential. Treatment methods for the removal of anionic perfluorochemical (PFC) surfactants from industrial effluents are needed to minimize the environmental release of these pollutants. Removal of PFC surfactants from aqueous solutions by sorption onto various types of granular activated carbon was investigated. Three anionic PFC surfactants, i.e., perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA) and perfluorobutane sulfonate (PFBS), were evaluated for the ability to adsorb onto activated carbon. Additionally, the sorptive capacity of zeolites and sludge for PFOS was compared to that of granular activated carbon. Adsorption isotherms were determined at constant ionic strength in a pH 7.2 phosphate buffer at 30 degrees C. Sorption of PFOS onto activated carbon was stronger than PFOA and PFBS, suggesting that the length of the fluorocarbon chain and the nature of the functional group influenced sorption of the anionic surfactants. Among all adsorbents evaluated in this study, activated carbon (Freundlich K(F) values=36.7 60.9) showed the highest affinity for PFOS at low aqueous equilibrium concentrations, followed by the hydrophobic, high-silica zeolite NaY (Si/Al 80, K(F)=31.8), and anaerobic sludge (K(F)=0.95-1.85). Activated carbon also displayed a superior sorptive capacity at high soluble concentrations of the surfactant (up to 80 mg l(-1)). These findings indicate that activated carbon adsorption is a promising treatment technique for the removal of PFOS from dilute aqueous streams. PMID- 18511100 TI - Comparison of the effectiveness of composting and vermicomposting for the biological stabilization of cattle manure. AB - Cattle manure is produced in large quantities in industrial breeding facilities and the storage and/or spreading of this waste on land may cause contamination of the atmosphere, soil and water. The aim of the present study was to evaluate the effectiveness of the active phases of composting, vermicomposting, and also a combination of composting and vermicomposting for reducing the polluting potential and for stabilizing cattle manure in the short-term. For this, the degree of decomposition as well as the microbial activity and microbial composition of the resulting products after the active phase of composting and vermicomposting were analysed. None of the treatments significantly reduced the dissolved organic carbon and dissolved organic nitrogen contents relative to the control, and therefore more time may be required for stabilization. Nevertheless, the lowest values of microbial biomass and activity corresponded to the earthworm worked substrates, in which fungal growth was also promoted; the combined treatment (composting + vermicomposting) was the most effective in terms of stabilizing the cattle manure. Moreover, earthworms promoted the retention of nitrogen and gradual release of P, as well as a reduction in electrical conductivity, thereby producing improved substrates for agricultural use. PMID- 18511101 TI - Two-generation toxicity study on the copepod model species Tigriopus japonicus. AB - Previous studies on the intertidal copepod Tigriopus japonicus have demonstrated that it is a suitable model species for the assessment of acute toxicities of marine pollutants. In order to standardize T. japonicus for use in environmental risk assessment involving whole life cycle exposure, we tested nine pollutants for their effects on growth and reproduction during a two-generation life cycle exposure test. Nauplii (F 0) were exposed to a range of concentrations of each chemical in a static renewal culture system. Broods of the second generation (F1) were subsequently exposed to the same concentrations for one full life cycle. Of the seven traits (nauplius phase, development time, survival, sex ratio, number of clutch, nauplii per clutch and fecundity), only the length of the nauplius phase and development time showed a greater sensitivity to chemical exposure. Between the two sensitive traits, the period of the nauplius phase was more sensitive than cohort generation time. Biocides significantly increased the maturation period of nauplii as well as copepodids in F 0 generation. In this study, it was demonstrated that T. japonicus could also be used in reproduction and life cycle tests and it provides an opportunity for testing the chronic and subchronic toxic effects of marine pollutants. Further validation and harmonization in a multi-centric study involving other laboratories of the region will strengthen its use as a supplement to existing model species. PMID- 18511102 TI - Effects of cold-dark storage on growth of Cylindrotheca closterium and its sensitivity to copper. AB - Cylindrotheca closterium cells were maintained at low temperature (4+/-1 degrees C) and dark conditions up to 21 weeks to assess the effect on survival and physiological status. From a control culture under standard conditions, three densities were prepared: (A) 2 x 10(4), (B) 10 x 10(4), and (C) 25 x 10(4) cells ml(-1). Weekly, inoculums of each stored density were exposed to continuous light and at 20+/-1 degrees C. Sensitivity to copper for microalgal cultures was evaluated in order to assess possible changes in cells sensitivity due to storage. Concurrently, assays with a control culture were carried out in order to assess the sensitivity of C. closterium to copper and to be able to generate a standard sensitivity control chart with a mean value of EC50-72 h+/-2SD (standard deviation). Density-C presented higher cell yield values, between 40% and 80% relative to control culture. Cell density showed to be important feature that may be taken into account in cell storage experiments. There was an increase in sensitivity of cells submitted to storage; however results always kept in the range established as standard sensitivity with no statistically significant difference with regards to control culture. EC50-72 h mean value for the control culture was 29+/-10 mug Cul(-1), while for densities-A, B and C were 22+/-7; 23+/ 9 and 23+/-8 microg Cul(-1), respectively. In spite of drastic changes in the environmental conditions due to storage, it is concluded that C. closterium cells stored during 5 months remained metabolically active and with no significant change in its sensitivity. PMID- 18511103 TI - Total TEQ reference range (PCDDs, PCDFs, cPCBs, mono-PCBs) for the US population 2001-2002. AB - We report reference ranges for the total toxic equivalency (TEQ) and TEQ sub fractions of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), coplanar biphenyls (cPCBs), and mono-ortho-substituted biphenyls (mPCBs) in a statistically designed sampling of the US population in 2001-2002. The TEQ and TEQ sub-fractions have been stratified by age, sex, and race/ethnicity. The TEQ levels are lower using the 2005 toxic equivalency factors (TEFs) compared to using the 1998 TEF values, principally due to the much lower 2005 TEF values assigned to the mPCBs. Mexican Americans (MA) have significantly lower TEQ levels than both non-Hispanic whites (NHW) and non-Hispanic blacks (NHB). Using the 1998 or 2005 TEF values, males and females have nearly the same distribution of TEQ sub-fractions. We found a significant increase in TEQ levels with age for males, females, and NHW. About 80-90% of the total TEQ can be estimated by using seven congeners, namely 2,3,7,8-TCDD, 1,2,3,7,8-PeCDD, 1,2,3,6,7,8-HxCDD, 2,3,4,7,8 PeCDF, PCB-126, PCB-118, and PCB-156. We also measured geometric mean TEQ levels in pooled samples from the US population. The geometric mean TEQ levels also increase with age. In the youngest age group (12-19 years), the TEQ levels were higher in males than in females while females had higher TEQ levels than males in all older age groups. In the pools, as age increases the percent contribution of the PCDD TEQ levels increases while the percent contribution of the PCDF TEQ levels decreases for all race/ethnicity and sex strata. PMID- 18511104 TI - Disruption of hepatic mitochondrial bioenergetics is not a primary mechanism for the toxicity of methoprene - relevance for toxicological assessment. AB - Methoprene (isopropyl(2E,4E)-11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate) is an insect growth regulator generally used to control insect populations by preventing insect maturation. So far, the effects of the insecticide on mitochondrial bioenergetics were not investigated. In the present work, liver mitochondria from Wistar rats were isolated and features of mitochondrial physiology were characterized in the presence of methoprene. High concentrations of methoprene, in the range of 40-100 nmol/mg of protein could decrease the transmembrane electric potential (Delta Psi) developed by mitochondria and, at the highest concentration, methoprene prevented complete Delta Psi repolarization after ADP addition. The effect was more evident using succinate than with ascorbate+TMPD as substrate. State 3 respiration was approximately 60% inhibited by 80 nmol of methoprene/mg of protein, while state 4 respiration, within the same range of methoprene concentrations, showed a slight increase, when both glutamate-malate and succinate were used as substrates. Additionally, FCCP stimulated respiration was inhibited to an extent comparable to the effect on state 3, which suggests an interaction of methoprene with the respiratory chain, more evident with glutamate/malate as substrate. The activity of complex I (NADH ubiquinone oxidorreductase) and that of the segment comprehending complexes II and III (succinate-cytochrome c reductase) were decreased in the presence of methoprene (approximately 60% and 85% of inhibition, respectively, with 300 nmol of methoprene/mg of protein), while the activities of cytochrome c oxidase and ATPase do not seem to be affected. Furthermore, the action of methoprene on the mitochondrial permeability transition was also studied, showing that the insecticide (in the range of 30-80 nmol mg(-1) of protein) decreases the susceptibility of liver mitochondria to the opening of the transition pore, even in non-energized mitochondria. These results lead to the conclusion that methoprene interference with hepatic mitochondrial function occurs only for high concentrations, which implies that the noxious effects of the insecticide reported for a number of non-target organisms are not fully attributable to mitochondrial effects. Therefore, it seems that mitochondrial activity does not represent the primary target for methoprene toxic action. PMID- 18511105 TI - Effect of manganese ion on the mineralization of 2,4-dichlorophenol by ozone. AB - Mineralization of 2,4-dichlorophenol (DCP) was studied by ozone with Mn(2+) as an ozonation catalyst. Laboratory scale semi-batch ozonation experiments were conducted at room temperature. The results showed that trace amount of Mn(2+) accelerated the mineralization of DCP. Total organic carbon removal rate was independent on Mn(2+) dosage at its range of 0.1-0.5 mgL(-1). Dissolved ozone concentration in the solution remained low level in the catalytic ozonation process, which indicated that Mn(2+) catalyzed decomposition of ozone. DCP mineralization was inhibited in catalytic ozonation by the addition of carbonate. Electron spin resonance/spin-trapping technique was used to determine hydroxyl radicals, and the results showed that larger amounts of hydroxyl radicals were produced in catalytic ozonation system than those of single ozonation. Intermediates mainly including aliphatic carboxylic acids were determined qualitatively and semi-quantitatively by GC-MS. And, a general pathway for mineralization of DCP was proposed. PMID- 18511107 TI - Spatial and temporal variation of particle number concentration in Augsburg, Germany. AB - Epidemiological studies on health effects of outdoor air pollution are largely based on a single monitoring site to estimate the exposure of people living in urban areas. For such an approach two aspects are important: the temporal correlation and the spatial variation of the absolute levels of concentrations measured at different sites in an urban area. Whereas many studies have shown small spatial variability of fine particles in urban areas, little is known on how well a single monitoring station could represent the temporal and spatial variation of ultrafine particles across urban areas. In our study we investigated the temporal and spatial variation of particle number concentration (PNC) at four background sites in Augsburg, Germany. Two of them were influenced by traffic, one was placed in the outskirts of the city. The average PNC levels at two urban background sites with traffic impact were 16,943 cm(-3) and 20,702 cm(-3), respectively, compared to 11,656 cm(-3) at the urban background site without traffic impact (ratio 1.5 to 1.8). The Spearman correlation coefficients between the monitoring sites were high (r>0.80). The pronounced differences in absolute PNC levels suggest that the use of a single monitoring station in long-term epidemiological studies must be insufficient to attribute accurate exposure levels of PNC to all study subjects. On the other hand, the high temporal correlations of PNC across the city area of Augsburg implicate that in epidemiological time-series studies the use of one single ambient monitoring site is an adequate approach for characterizing exposure to ultrafine particles. PMID- 18511106 TI - Haematological and biochemical effects of polyphenolics in animal models. AB - Polyphenols of natural and synthetic origin are exploited in tanning sector to convert putrescible skin/hide to non-putrescible leather. However, only 30-40% of the inputs have been taken up for processing, the remaining is released as unspent. The existing conventional wastewater treatment systems are inefficient in removing or degrading these unspent polyphenols and thus detrimental to ecosystem. The present study demonstrates the evaluation of impact of both synthetic and natural polyphenols on biochemical and haematological properties of blood and serum in animal models. The results reveal that concentrations of polyphenols play a major role. At higher concentrations, irrespective of their nature, there was a marked change in the lipid profile (81% reduction), followed by insignificant change in glucose levels, RBC and WBC counts and other haematological parameters. At lower concentrations, no significant changes in the above said properties were observed. PMID- 18511108 TI - Assessing urinary levoglucosan and methoxyphenols as biomarkers for use in woodsmoke exposure studies. AB - A major contributor to particle concentrations in urban airsheds is domestic woodsmoke and smoke arising from wildfires or management burns. Particle concentrations in urban airsheds have been associated with a wide range of health effects. There has been little research into the contribution of biomass burning to studies of human health due to the complexity of attributing effects in the presence of multiple sources of pollutants and the variability in the nature and conditions of biomass burning. A significant advance is the use of biomarkers of exposure; methoxyphenol and levoglucosan; specific compounds produced following the combustion of lignins and detected in urine. Levoglucosan has not previously been assessed for its usefulness as a marker of human exposure. We report for the first time levoglucosan concentrations in urine. Twelve participants were recruited and asked to provide spot urine samples pre- and post-exposure to a fire training exercise. Both levoglucosan and methoxyphenol were detected in the urine of participants. There was no significant increase in these compounds post exposure to smoke arising from the fire training. Further work is required to assess this biomarker for human exposure studies and in particular the role of diet and previous exposure. PMID- 18511109 TI - Pilot study of homocysteine and cysteine in patients with thrombosis in different vascular sites. Epidemiology and response to folate. AB - Hyperhomocysteinemia is a risk factor for arterial and venous thrombosis. However, lowering homocysteine (Hcy) with vitamins not only failed to improve outcomes but also may lead to recurrent events. Our objectives were to evaluate Hcy and cysteine (Cys) levels in patients with thrombosis in different vascular sites, and their response to folate. One hundred and sixty four consecutive patients with thrombosis (42.1% arterial (AT), 36% venous (VT), 4.9% both venous and arterial thrombosis (AVT) and 17% unusual site (UST)) were included. Hcy and Cys were highest in patients with AVT and UST (p=0.0006). Ninety-three patients were treated, 70% were followed-up. Hcy levels normalized after therapy in all patients. Cys levels tended to vary after therapy according to the site of thrombosis. We observed a significant correlation between folate and Hcy (r: 0.48; p=0.005) among homozygous for MTHFR. A significant inverse relation was observed between Hcy and folate among homozygous and heterozygous (r: 0.462, p=0.007 and r: 0.267; p=0.04, respectively). No correlation was observed between folate and Cys. In conclusion, our observations suggest that Hcy and Cys might be implicated in thrombosis in different vascular sites, and respond differently to folate. PMID- 18511110 TI - Luteinized thecoma associated with sclerosing peritonitis--conservative surgical approach followed by corticosteroid and GnRH agonist treatment--a case report. AB - BACKGROUND: Luteinized thecoma of the ovary associated with sclerosing peritonitis is a rare tumor that has no standard definitive treatment regimen. CASE: A 25 year-old patient diagnosed with luteinized thecoma and sclerosing peritonitis in the omentum. The patient received high dose corticosteroids (IV Hydrocortisone 500 mg/d) and GnRH agonist (IM Leuprolide 3.75 mg) in order to achieve ovarian suppression and relief of the clinical peritonitis. She was re admitted two weeks later due to bowel obstruction which was treated conservatively. The steroid regimen was continued by oral intake for 5 weeks with complete remission of the peritonitis related symptoms. The bilateral enlarged ovarian tumor-like solid was the prominent finding in consecutive ultrasound exams with no decrease in size despite of the above mentioned protocol. Thus, the patient was re-operated for exploration and biopsies of the ovary and the pathology report showed no evidence of remnant disease in the ovary, or in the peritoneum. Completing follow-up of 15 months since the last operation, the patient is asymptomatic. She conceived spontaneously and currently is in her 24th week of a normal pregnancy. CONCLUSION: This is the first case report in the English literature of a successful medical conservative treatment of a young patient with luteinized thecoma associated with sclerosing peritonitis that led to complete relief of the symptoms and allowed fertility preservation. PMID- 18511111 TI - Eugenol functionalized poly(acrylic acid) derivatives in the formation of glass ionomer cements. AB - Eugenol possesses analgesic and anti-inflammatory properties with the ability to relieve pain in irritated or diseased tooth pulp, thus, incorporating polymers with eugenol moieties in dental cements is attractive. An acrylic derivative of eugenyl methacrylate (EgMA) was copolymerized with acrylic acid (AA) using a radical initiator, to yield a water soluble copolymer of acrylic acid and eugenyl methacrylate {p(AA-co-EgMA)}, which was then applied in the formulation of glass ionomer cements for potential application as dental cements. Three concentrations of the p(AA-co-EgMA) copolymer in water were studied by, 30wt%, 40wt% and 50wt%, and used with different powder:liquid ratios to formulate the glass-ionomer cements. The setting kinetics showed that both the concentration of the copolymer and the powder:liquid ratio influenced the working and setting times. Thus, selected formulations were used for further characterization of their mechanical properties, water uptake and fluoride release, to optimize the cement formulation. The experimental glass-ionomer cements exhibited physical and mechanical properties in compliance to ISO standard requirements with the benefit of the initial pH being greater than the commercial formulation used as the standard cement. Furthermore, the presence of the eugenyl moieties bound to the polymer matrix was advantageous with respect to moisture sensitivity and anti bacterial properties. PMID- 18511112 TI - The International Conference on Chitin and Chitosan. PMID- 18511113 TI - Successful treatment with VAD of a myelodysplastic syndrome occurring during the course of a smoldering multiple myeloma. PMID- 18511114 TI - The effects of community violence on children in Cape Town, South Africa. AB - OBJECTIVE: The primary objective of the study was to investigate the relationship between exposure to community violence (neighborhood, school, police, and gang violence) and psychological distress in a sample of children living in the Cape Town, South Africa area. Another objective was to identify variables that moderate and mediate the relationship between exposure to community violence and psychological distress. METHODS: Face-to-face interviews were conducted with 185 children between the age of 8 and 13 from five Cape Town Township schools. Structured scales were used to measure exposure to several forms of community violence, family functioning, social support, perceptions of safety, and "unknown" locus of control. RESULTS: Exposure to all forms of violence was extremely high and resulted in substantial psychological distress. Perceived safety functioned as a mediating variable for all forms of violence. Unknown locus of control, social support, family organization, and family control moderated the effects of exposure to certain kinds of violence. Surprisingly, exposure to murder was not related to psychological distress, suggesting a possible "numbing" effect of extreme forms of violence. Hearing about violence from others had almost the same effect as actually witnessing it. Older children had witnessed more violence and were experiencing more distress, suggesting an "exposure accumulation" effect. CONCLUSIONS: The findings suggest the importance of a child's ability to feel safe in reducing the distress that occurs as a result to exposure to violence. Parents and schools can help children cope, but there appear to be limits. Early intervention, before maladaptive coping mechanisms have developed, also appears to be important. PMID- 18511115 TI - Exploration and validation of clusters of physically abused children. AB - OBJECTIVE: Cluster analysis was used to enhance understanding of heterogeneity in social adjustment of physically abused children. METHOD: Ninety-eight physically abused children (ages 5-10) were clustered on the basis of social adjustment, as measured by observed behavior with peers on the school playground and by teacher reports of social behavior. Seventy-seven matched nonabused children served as a comparison sample. Clusters were validated on the basis of observed parental sensitivity, parents' self-reported disciplinary tactics, and children's social information processing operations (i.e., generation of solutions to peer relationship problems and attributions of peer intentions in social situations). RESULTS: Three subgroups of physically abused children emerged from the cluster analysis; clusters were labeled Socially Well Adjusted, Hanging in There, and Social Difficulties. Examination of cluster differences on risk and protective factors provided substantial evidence in support of the external validity of the three-cluster solution. Specifically, clusters differed significantly in attributions of peer intent and in parenting (i.e., sensitivity and harshness of parenting). Clusters also differed in the ways in which they were similar to, or different from, the comparison group of nonabused children. CONCLUSIONS: Results supported the contention that there were clinically relevant subgroups of physically abused children with potentially unique treatment needs. Findings also pointed to the relevance of social information processing operations and parenting context in understanding diversity among physically abused children. PRACTICE IMPLICATIONS: Pending replication, findings provide support for the importance of considering unique treatment of needs among physically abused children. A singular approach to intervention is unlikely to be effective for these children. For example, some physically abused children might need a more intensive focus on development of prosocial skills in relationships with peers while the prosocial skills of other abused children will be developmentally appropriate. In contrast, most physically abused children might benefit from training in social problem-solving skills. Findings also point to the importance of promoting positive parenting practices in addition to reducing harsh discipline of physically abusive parents. PMID- 18511117 TI - Mediational significance of PTSD in the relationship of sexual trauma and eating disorders. AB - OBJECTIVE: To examine the mediational significance of posttraumatic stress disorder (PTSD) and the development of eating disorder symptomatology following sexually traumatic experiences. METHOD: Seventy-one victims of sexual trauma and 25 control subjects completed interviews and questionnaires assessing eating disorder psychopathology and posttraumatic stress disorder symptomatology. Mediational analyses were conducted examining the relationships among trauma, posttraumatic stress, and eating disorder symptoms. Mediational significance was assessed by the drop in the overall correlation between trauma and eating disorder symptoms when PTSD symptoms were included in the regression model. RESULTS: There is a significant association between a history of trauma and eating disorder symptoms. Also, there was a significant association between a history of trauma and posttraumatic stress disorder symptoms. Importantly, the relationship between trauma and eating disorder symptoms was significantly reduced when posttraumatic stress disorder symptoms were included in the regression analyses, indicating mediational significance of the posttraumatic stress construct. These findings were most pronounced for the physiological arousal and avoidance components of posttraumatic stress disorder. DISCUSSION: The present findings support the idea that individuals who develop eating disorders after sexual trauma are likely to have experienced posttraumatic stress disorder symptomatology. These findings have significant implications for causal models of eating disorder onset in trauma victims. Furthermore, clinical interventions for traumatized eating disordered individuals may benefit from a focus on posttraumatic stress symptomatology. PMID- 18511116 TI - Protective and vulnerability factors for physically abused children: effects of ethnicity and parenting context. AB - OBJECTIVE: Although social maladjustment appears to be common among abused children, negative outcomes are not inevitable. This investigation was designed to determine whether ethnicity and features of the parenting context predicted children's social adjustment, and whether the strength and direction of these relations differed for abused and nonabused children. METHOD: Participants included 78 physically abused and 75 demographically matched nonabused children and one of their parents. Observations of parenting were used to measure parental sensitivity, and parent self-reports of depression were obtained using the SCL-90 R. Children's peer social adjustment was measured by teacher report. RESULTS: Using regression analysis, we tested whether each potential protective or vulnerability factor interacted with abuse status in prediction of social adjustment. Results indicated main effects of ethnicity and sensitivity for prosocial behavior, and a main effect of sensitivity for aggression. In addition, there was a significant interaction of ethnicity and abuse status for aggression such that there was a significant difference between abused and nonabused European American children but not between abused and nonabused African American children. CONCLUSIONS: Findings indicate that risk for aggressive behavior among abused children might be culturally specific rather than universal. In addition, results point to beneficial effects of parental sensitivity for maltreated children. PMID- 18511118 TI - Developmental experiences of child sexual abusers and rapists. AB - OBJECTIVE: The aim of this study is to identify the distinct developmental experiences associated with child sexual abuse and rape. METHOD: For 269 sexual offenders (137 rapists and 132 child sexual abusers), developmental experiences were recorded from a behavioral checklist, a parental-bonding survey, and a sexual history questionnaire. Offender classification was obtained from official records and verified through polygraph examinations. RESULTS: Compared to rapists, child sexual abusers reported more frequent experiences of child sexual abuse (73%), early exposure to pornography (65% before age 10), an earlier onset of masturbation (60% before age 11), and sexual activities with animals (38%). In contrast to child sexual abusers, rapists reported more frequent experiences of physical abuse (68%), parental violence (78%), emotional abuse (70%), and cruelty to animals (68%). Both child sexual abusers and rapists (>93%) reported frequent exposure to violent media during their childhood. Most offenders (94%) described having insecure parental attachment bonds; 76% of rapists reported avoidant parental attachments and 62% of child sexual abusers reported anxious parental attachments. CONCLUSIONS: Findings from this study support the role of specific developmental experiences as etiological factors in differential sexual offending. Child sexual abusers' developmental histories were characterized by heightened sexuality; whereas rapists' childhood histories were more indicative of violence. These findings have implications for the treatment of sexual abusers and the prevention of sexual abuse. PRACTICE IMPLICATIONS: This study's findings suggest that sexual offenders have been socialized to satisfy human needs of intimacy and sexuality through maladaptive means, which implies that a risk management approach may not be sufficient treatment. Although risk models teach offenders skills to avoid high-risk situations, they fail to address the maladaptive strategies that they may have developed for satisfying needs. Instead, the focus of treatment should be to equip offenders with the knowledge, skills, and opportunities to achieve these needs in an acceptable manner. Thus, this model will provide these individuals with the opportunity to live a healthy life without sexual offending. PMID- 18511119 TI - Interaction between E-protein and Oct transcription factors in the function of the catfish IGH enhancer. AB - Transcriptional control of the immunoglobulin heavy chain (IGH) locus in the channel catfish, Ictalurus punctatus, is incompletely understood. It is, however, known that 2 variant octamer motifs and a microE5 motif in the core region of the enhancer (Emicro3') are important in driving transcription, and it has been suggested that interaction between transcription factors (Oct factors and E proteins) bound to these sites contributes to enhancer function. In this study, the functional relationships between the microE5 motif, the proximal octamer motif, and the factors that bind them have been examined. The results of mutational analysis of these motifs showed that their interaction is important to driving transcription from the enhancer. Furthermore, the catfish Oct transcription factors were capable of a physical interaction with the catfish E proteins. These results support a role for interaction between transcription factors bound to the octamer and microE5 motifs in the function of the Emicro3' enhancer. PMID- 18511120 TI - Complete sequence of Enterococcus faecium pVEF3 and the detection of an omega epsilon-zeta toxin-antitoxin module and an ABC transporter. AB - Glycopeptide resistant Enterococcus faecium (GREF) persists on Norwegian poultry farms despite the ban on the growth promoter avoparcin. The biological basis for long-term persistence of avoparcin resistance is not fully understood. This study presents the complete DNA sequence of the E. faecium R-plasmid pVEF3 and functional studies of some plasmid-encoded traits (a toxin-antitoxin (TA) system and an ABC transporter) that may be of importance for plasmid persistence. The pVEF3 (63.1 kbp), isolated from an E. faecium strain of poultry origin sampled in Norway in 1999, has 71 coding sequences including the vanA avoparcin/vancomycin resistance encoding gene cluster. pVEF3 encodes the TA system omega-epsilon-zeta, and plasmid stability tests and transcription analysis show that omega-epsilon zeta is functional in Enterococcus faecalis OGIX, although with decreasing effect over time. The predicted ABC transporter was not found to confer reduced susceptibility to any of the 28 substances tested. The TA system identified in the pVEF-type plasmids may contribute to vanA plasmid persistence on Norwegian poultry farms. However, size and compositional heterogeneity among E. faecium vanA plasmids suggest that additional plasmid maintenance systems in combination with host specific factors and frequent horizontal gene transfer and rearrangement causes the observed plasmid composition and distribution patterns. PMID- 18511121 TI - Projective risk variables in early adolescence and subsequent disinhibitory psychopathology. AB - The objective was to examine early adolescent projective risk indicators for the development of antisocial behaviour as related to adult personality traits, psychopathy, and violent behaviour over the life span. Assessment data included Rorschach (Rr) ratings (at age 11-14 years), personality inventories (EPQ-I and KSP scales), and a shortened Psychopathy Check List (PCL) (administered at age 32 40 years), obtained from a group of 199 male subjects; and smoking habits (at age 36-44 years) obtained from 125 of those subjects. Results, controlled for intelligence, indicated that the high and very high risk groups, as determined by level of total Rr risk scores, were (1) significantly higher on self-rated IVE Impulsiveness, the anxiety-related KSP Muscular Tension, and nonconformity traits, as compared to the low Rr risk group--the very high risk group also scoring significantly higher on the EPQ Psychoticism scale, related to aggressiveness and cruelty; (2) higher on clinically rated PCL total sum and factor scores; and (3) they were overrepresented among Ss with subsequent violent offence, and Ss with heavy smoking habits. The results are discussed in terms of the possible usefulness of psychodynamic oriented cognitive-emotional indicators in the search for underlying mechanisms in the development of disinhibitory psychopathology. PMID- 18511122 TI - Multifactorial origin of high incidence of Serratia marcescens in a cardio thoracic ICU: analysis of risk factors and epidemiological characteristics. AB - OBJECTIVE: A four-fold increase in the incidence of Serratia marcescens occurred in a cardio-thoracic ICU within a 13-month period. Clinical, epidemiological and molecular characteristics were analysed to elucidate the outbreak's origin. METHODS: Epidemiological data were analysed by mapping clustered cases; isolates were genotyped by AFLP analysis. A case-control study was performed to identify risk factors for the acquisition of S. marcescens. Data were obtained from files and electronic databases of the ICU and Department of Medical Microbiology. The adherence to hygiene protocols on the ICU was reviewed by a medical audit. RESULTS: Genotyping showed 16 distinct S. marcescens strains. Twenty-one cases and 39 controls were enrolled in the case-control study. Significant differences found by univariate analysis included the duration of surgery, APACHE-II-score on ICU admission, length of ICU stay, duration of mechanical ventilation, tube feeding and the sum of the number of days per invasive device. In a multivariate logistic regression model, the length of ICU stay and tube feeding were independent risk factors. Outbreak strains were not more frequently resistant to gentamicin, ciprofloxacin, meropenem or trimethoprim-sulfamethoxazole as compared to a reference group. Hygiene protocols, including hand washing, were insufficiently practiced by the ICU's medical staff. CONCLUSIONS: The heterogeneity of the strains points to transmission from various sources. This outbreak of S. marcescens was most probably caused by reduced hand washing and other breaks in infection prevention protocols in combination with the presence of the identified risk factors, which act by affecting the number and intensity of potential transmission events. PMID- 18511123 TI - Physical growth delays and stress dysregulation in stunted and non-stunted Ukrainian institution-reared children. AB - To study the effect of institutional rearing on physical growth and stress regulation we examined 16 institution-reared children (3-6 years old) in Ukraine and compared them with 18 native family-reared children pair-matched on age and gender. Physical growth trajectories were examined on the basis of archival medical records and current measurements of height, weight, and head circumference. Stress regulation was studied on the basis of diurnal salivary cortisol sampled six times during 1 day. 31% of institution-reared children were stunted at 48 months whereas none of the family-reared children were. Substantial delays in physical growth were observed in institution-reared children especially during the first year of life. From 24 months onwards a tendency for improvement in physical growth was evident among the temporarily stunted institution-reared children, with complete catch-up in weight and partial catch-up in height by the time of assessment. Chronically stunted institution-reared children demonstrated persistent severe growth delays. Institution-reared and family-reared children showed similar patterns of diurnal cortisol production with decreases over the day. However, temporarily stunted institution-reared children had a significantly higher total daily cortisol production than both chronically stunted institution reared children and family-reared children. These data confirm previous findings regarding physical growth delays and stress dysregulation associated with institutional care, but also point to differences in cortisol production between stunted and non-stunted institution-reared children. PMID- 18511124 TI - Adaptive properties and heterogeneity of dopamine D(2) receptors - pharmacological implications. AB - In this review, we focus on the marked adaptability of dopamine D(2) receptors to varying agonist levels and we discuss the extent to which this phenomenon can account for the heterogeneity of these receptors in regard to function and pharmacological responsiveness. We emphasize the significance of a distinction between synaptic and extrasynaptic receptors in this context. For example, the application of this dichotomy appears to shed new light on the various subgroups of antipsychotic drugs and the mechanisms underlying their different profiles. PMID- 18511125 TI - The development of a novel automated taste stimulus delivery system for fMRI studies on the human cortical segregation of taste. AB - fMRI indicated that the primary taste cortex is activated not only by taste but also by non-taste information from oral stimuli. Head movements caused by swallowing are very critical problem in fMRI and inherent difficulties to modulate taste stimuli in the mouth exist to elucidate functional segregation of human brain. We developed a novel automated taste stimulus delivery system for fMRI studies to segregate the pure taste area in the primary taste cortex in humans. As a novel intra-oral device, an elliptic cylinder was attached to an individual mouthpiece and then subject placed the tongue tip in it. Using a computer-controlled extra-oral device, the solutions ran through the intra-oral device in constant conditions. Three adult volunteers participated in the experimental session, alternately consisting of 30 pairs of taste stimuli (0.5 mol/l sucrose solution) and control (water) blocks. The typical findings of the three subjects revealed activation only in the primary taste cortex (P<0.001), and none in the secondary taste cortex. This is the first system that delivers the taste stimuli automatically to a standardized area on the subject's tongue under constant conditions, thus allowing us to successfully segregate the pure taste area in the primary taste cortex in humans. PMID- 18511126 TI - Real-time control of stepper motors for mechano-sensory stimulation. AB - Mechanical stimulation is widely used to study sensory encoding in the nervous system of living organisms. The stimulation of mechano-receptor neurons is achieved through a large variety of devices that generate movement or vibration. In many situations, a hard real-time (RT) control of the device (in the millisecond time scale) is needed to produce realistic mechanical stimuli. The real-time control can be required to achieve the desired precision in the device or to implement activity-dependent stimulation protocols that imply the detection of physiological events to drive the stimulus in real time. In this paper we show that real-time software technology can be used to control stepper motors for mechano-receptor stimulation, and to implement artificial closed-loops to address the sensory-motor transformation. We illustrate this using as an example the control of a stepper motor to precisely move gravimetric organs in in vitro preparations. PMID- 18511127 TI - A robust and accurate algorithm for estimating the complexity of the cortical surface. AB - A fractal dimension (FD) gives a highly compact description of the shape characteristics of the human brain and has been employed in many studies on brain morphology. The accuracy of FD estimation depends on the precision of the input shape description. Facilitated by automatic cerebral cortical surface reconstruction algorithms, the shape of the cerebral cortex can be more precisely modeled using Magnetic Resonance (MR) imaging. Since the reconstructed cortical surface is represented by triangles, rather than by points, as is typical of models that use voxels, the voxel-based FD estimation algorithms that have been used in previous studies do not work when using the cortical surface as the input. Thus, designing a new algorithm that is able to estimate the FD from a surface representation becomes of particular interest. In this paper, a robust and accurate FD estimation algorithm is proposed. The algorithm is based on a box triangle intersection checking strategy, which is used for the first time in brain analyses, and a box-counting method, which has been widely used in FD computations of the human brain and other natural objects. These two features endowed the algorithm with robustness. The accuracy of the algorithm was validated via several experiments using both manually generated datasets and real MR images. As a result of these features, the algorithm is also suitable for estimating the FD of fractals in addition to that of the cerebral cortex. PMID- 18511128 TI - Long-range correlation of renal sympathetic nerve activity in both conscious and anesthetized rats. AB - In this study we employed both detrended fluctuation analysis (DFA) and multiscale entropy (MSE) measurements to compare the long-range temporal correlation (LRTC) of multifibre renal sympathetic nerve activity (RSNA) between conscious and anesthetized Wistar rats. It was found that both methods showed the obvious LRTC properties in conscious state. Moreover, the scaling exponent of the RSNA in conscious rats was significantly higher than that in anesthetized rats. The results of MSE analysis showed that the entropy values, derived from the conscious group, increased on small time scales and then stabilized to a relatively constant value whereas the entropy measure, derived from anesthetized animals, almost monotonically decreased. This suggests that the fractal properties of underlying dynamics of the system have been reduced by anesthesia. The results demonstrate that apparently random fluctuations in multifibre RSNA are dictated by a complex deterministic process that imparts "long-term" memory to the dynamic system. However, this memory is significantly weakened by anesthesia. PMID- 18511129 TI - Attrition factors in clinical trials of comorbid bipolar and substance-related disorders. AB - BACKGROUND: This study analyzed and defined specific factors that account for attrition in clinical research for patients with bipolar and substance-related disorders. METHODS: Data were analyzed from two completed studies: an open-label trial of lamotrigine in patients with bipolar disorder (BPD) and cocaine-related disorder, and a placebo-controlled trial of quetiapine in patients with BPD and alcohol-related disorders. Correlations and Independent sample t-tests were performed to assess the impact of baseline characteristics including on length of study participation. Significance was set at the p=0.05 level. RESULTS: In the lamotrigine-treated patients, the presence of an amphetamine-related disorder, in addition to cocaine-related disorders, was associated with a shorter time in the study. In the quetiapine-treated patients higher scores on the Addiction Severity Index Legal subscale were associated with shorter length in the study. The presence of panic disorder was associated with shorter time in both studies. LIMITATIONS: Although the data were taken from the two largest clinical trials, to date, in patients with BPD and substance-related disorders, the sample sizes were relatively modest. In addition, the baseline assessments were somewhat different in the two studies limiting our ability to make conclusions on differences between patients with BPD and cocaine use versus alcohol use. CONCLUSIONS: This study adds to an emerging literature on the significance of panic disorder in patients with BPD. PMID- 18511130 TI - Complex PTSD, interpersonal trauma and relational consequences: findings from a treatment-receiving Northern Irish sample. AB - BACKGROUND: The relationship between PTSD and complex PTSD remains unclear. As well as further addressing this issue, the current study aimed to assess the degree to which DESNOS (complex PTSD) was related to interpersonal trauma and had relational consequences. METHODS: Eighty one treatment-receiving participants with a history of exposure to the 'Troubles' in Northern Ireland, were assessed on various forms of interpersonal trauma, including exposure to the Troubles, and measures of interpersonal and community connectedness. RESULTS: DESNOS symptom severity was related to childhood sexual abuse and perceived psychological impact of Troubles-related exposure. A lifetime diagnosis of DESNOS was related to childhood Troubles-related experiences, while a current diagnosis of DESNOS was associated with childhood emotional neglect. PTSD avoidance predicted current DESNOS diagnosis and severity. Feeling emotionally disconnected from family and friends (i.e., interpersonal disconnectedness) was related to all three indices of DESNOS (i.e., lifetime diagnosis, current diagnosis and current symptom severity). LIMITATIONS: Sample characteristics (i.e., treatment-receiving) and size may limit the generalizability of findings. CONCLUSIONS: Complex PTSD is associated with PTSD but when present should be considered a superordinate diagnosis. PMID- 18511131 TI - Estrogen receptor gene 1 variants are not associated with suicidal behavior. AB - Estrogen is thought to be involved in the pathophysiology of depression and suicidal behaviors. We studied gene variants of estrogen receptor alpha (rs827421, rs1913474, rs1801132, rs722207, rs974276 and rs910416) in 167 German suicide attempters (affective spectrum n=107, schizophrenia spectrum n=35, borderline personality disorder n=25), 92 German individuals who committed suicide and 312 German healthy subjects. Single markers and haplotype analysis in relation to suicidal behaviors (suicide attempters/completers) did not reveal any significant association. These were also not associated with related features, such as violence or impulsivity of suicide attempt, State-Trait Anger Expression Inventory (STAXI) and Questionnaire for Measuring Factors of Aggression (FAF) scores. In conclusion, our study does not support the hypothesis that estrogen receptor alpha gene variants are major contributors to suicide or to anger- or aggression-related behaviors. PMID- 18511132 TI - Empathy, social functioning and schizotypy. AB - Whilst affective empathy is concerned with one's emotional response to the affective state of another, cognitive empathy refers to one's understanding of another's mental state, and deficits in both are believed to contribute to the social behavioral abnormalities associated with schizophrenia. The present study aimed to test whether individual differences in normally distributed schizotypal personality traits are related to cognitive and affective empathy, and whether any observed association between schizotypy and empathy mediates the relationship between schizotypy and (reduced) social functioning. Non-clinical volunteers (N=223) completed measures of schizotypal personality, cognitive and affective empathy, social functioning and negative affect. The results indicated that higher schizotypy was associated with reduced empathy, poorer social functioning and increased negative affect. Of the specific schizotypal dimensions (positive, negative and disorganized), only negative schizotypy was significantly associated with social functioning, and this relationship persisted even after controlling for negative affect. Further, affective empathy functioned as a partial mediator in this relationship. These data show that the relationship between negative schizotypy and social functioning is at least partially attributable to deficits in affective empathy. PMID- 18511133 TI - Immune responses of recombinant adenovirus co-expressing VP1 of foot-and-mouth disease virus and porcine interferon alpha in mice and guinea pigs. AB - Foot-and-mouth disease (FMD) is a highly contagious and economically devastating vesicular disease of cloven-hoofed animals. In this study, we constructed and characterized the immune responses and vaccine efficacy conferred by the recombinant adenovirus co-expressing VP1 of FMDV and porcine interferon alpha as fusion protein (rAd-pIFNalpha-VP1). Six groups of female BALB/c mice each with 18 were inoculated subcutaneously twice 2-week intervals with the recombinant adenoviruses. The results showed that the levels of humoral and cell-mediated immune responses in the group inoculated with rAd-pIFNalpha-VP1 were significantly higher than those in the group inoculated with rAd-VP1+rAd pIFNalpha (P<0.05). Then four groups of guinea pigs each with six were inoculated two times at 2-week intervals intramuscularly with rAd-pIFNalpha-VP1, commercial inactivated FMD vaccine, wild-type adenovirus (wtAd) or PBS, and the protective efficacy of rAd-pIFNalpha-VP1 was determined. The results indicated that all the guinea pigs vaccinated with rAd-pIFNalpha-VP1 as well as inactivated FMD vaccine were protected from FMDV challenge, even though the levels of neutralizing antibodies (1:32-1:40) of the animals vaccinated with rAd-pIFNalpha-VP1 was lower than that in the group inoculated with inactivated FMD vaccine (1:64-1:128). It demonstrated that the newly recombinant adenovirus rAd-pIFNalpha-VP1 might further be an attractive candidate vaccine for preventing FMDV infection in swine. PMID- 18511134 TI - Chemokines and chemokine receptors in the nervous system Rome, 27/28 October, 2007. PMID- 18511135 TI - A temporal analysis of the relationships between social stress, humoral immune response and glutathione-related antioxidant defenses. AB - The exposure to different kinds of stress impacts on the reactive oxygen species production with potential risk to the integrity of the tissues. Psychological or biological stress is responsible for a significant increase in the oxidative stress markers and also for activation of the antioxidant defense system. In this study, we analyzed the relationships between social stress, humoral immune response and glutathione-related antioxidant defenses. Groups of male Swiss mice were subjected to different lengths of social stress exposure (social confrontation) which varied from 1 up to 13 days. As a biological stressor, 10(9) sheep red blood cells (SRBC)/mL were injected by intraperitoneal route. As controls, animals not subjected to social stress and/or injected with vehicle solution were used. The serum samples and the cerebral cortex were collected at 4 h, 3, 5, 7, 9, 11, and 13 days after the end of social confrontation. The results indicated that the antioxidant enzymes activities were affected by psychological as well as by biological stressor. These alterations were dependent on the timing of stress exposure which resulted in a positive or in a negative correlation between the antibody titres to SRBC and antioxidant enzymes. We also discuss the possible role of SRBC injection in the modulation of the effects of psychosocial stress on antioxidant metabolism. PMID- 18511136 TI - Evidence for multiple mechanisms of toxicity in larval rainbow trout (Oncorhynchus mykiss) co-treated with retene and alpha-naphthoflavone. AB - Alkylated polycyclic aromatic hydrocarbons, such as retene (7-isopropyl-1 methylphenanthrene), induce cytochrome P450 1A (CYP1A) enzymes and produce dioxin like toxicity in the embryo-larval stages of fish characterized by the signs of blue sac disease (BSD). The signs of toxicity are well characterized; however, the mechanism is not well understood. To elucidate the role of CYP1A in retene toxicity, larval rainbow trout (Oncorhynchus mykiss) were co-treated with a range of concentrations of alpha-naphthoflavone (ANF), a known CYP1A inhibitor. The co treatment produced synergistic toxicity at 3.2-100 microg/L ANF, after which toxicity at 180 microg/L ANF dropped to levels typical of retene-only. At 320 microg/L ANF, toxicity increased with or without retene, indicating that ANF alone was capable of inducing BSD. In addition, the additive toxicity of retene only and 320 microg/L ANF-only approximately equalled that of the co-exposed larvae (100 microg/L retene+320 microg/L ANF), indicating response addition. Thus, two mechanisms of action occurred in co-exposed larvae at different concentrations of ANF. In trout larvae, there was a correlation between toxicity and CYP1A protein concentrations, and in juvenile trout, ANF produced a concentration-dependent inhibition of ethoxyresorufin-O-deethylase (EROD) activity without a measurable drop in CYP1A protein. Taken together, the mechanism underlying the synergistic toxicity is EROD-independent and may be AhR dependent. This study demonstrated that multiple, exposure-dependent mechanisms can occur in mixture toxicity, suggesting that current risk assessment models may drastically underestimate toxicity, particularly of mixtures containing both CYP1A inducers and inhibitors. PMID- 18511137 TI - Integrating individual ecdysteroid content and growth-related stressor endpoints to assess toxicity in a benthic harpacticoid copepod. AB - Anthropogenic chemicals released into the environment may have so-called endocrine disruptive effects. For instance, innumerous observations on subtle alterations of fish reproductive systems have been published in the scientific literature during the last decades. At the same time, the evidence is scarce regarding similar effects in crustaceans, which is probably related to the limited understanding of basic crustacean endocrine systems and pathways, rather than absence of endocrine disruption within the crustacean subphylum. This knowledge gap is particularly evident in micro-crustacean species, which are frequently used in environmental risk assessment of chemicals, and adequate tools for identification of potential endocrine disrupters are missing in current chemicals regulation. The main objective of the current study was therefore to utilize an enzyme immunoassay to establish a stable protocol for analysis of individual ecdysteroid levels in the benthic harpacticoid copepod Nitocra spinipes, a species which has been used in ecotoxicological investigations for more than 30 years. Further, to assess the usefulness of the individual ecdysteroid level as a stressor endpoint, it was integrated with a growth-related stressor endpoint battery, i.e. individual RNA content, mean development times, and growth rate, by exposing individual N. spinipes to the insecticide and known ecdysteroid antagonist lindane at 25-400 microg L(-1). The results showed that the ecdysteroid levels were significantly different from the control (71 pg individual(-1)) in the 100 microg L(-1) treatment (124 pg individual(-1)). Although the ecdysteroid levels were not significantly different from the control in the 25-50 microg L(-1) treatments (83-93 pg individual(-1)), these results still show a clear concentration-related trend. In the 200 microg L(-1) treatment, the ecdysteroid content was the highest (165 pg individual(-1)), however not significantly different from the control due to high variation. The 400 microg L(-1) treatment resulted in lowered ecdysteroid levels (107 pg individual(-1)), indicating a profound lindane-induced stress, which was confirmed by high mortality in the same treatment (79%). For all other endpoints there were clear concentration-related effects of lindane, with development time and growth rate as the most sensitive endpoints. In conclusion, this study presents for the first time a tool for identification of endocrine alterations in N. spinipes. By using the established enzyme immunoassay protocol, we obtained individual ecdysteroid levels that integrated well with the growth-related stressor endpoints previously used on this species. PMID- 18511138 TI - The effect of water accessibility on child health in China. AB - Using a dynamic panel model of child anthropometrics from China, the effect of an in-yard water source on child health was measured. Changes in within-community averages of household access to in-yard water were used as the instrument for changes in access to in-yard water sources. Further, to address the concern of non-random placements of water projects, correlations between changes in disease symptoms and community-level changes in access to in-yard water sources were examined. It was found that access to in-yard water sources improved child health only when mothers were relatively well educated. PMID- 18511139 TI - Auditory hallucinations and the mismatch negativity: processing speech and non speech sounds in schizophrenia. AB - BACKGROUND: In line with emerging research strategies focusing on specific symptoms rather than global syndromes in psychiatric disorders, we examined the functional neural correlates of auditory verbal hallucinations (AHs) in schizophrenia. Recent neuroimaging and behavioural evidence suggest a reciprocal relationship between auditory cortex response to external sounds versus that induced by AHs. METHODS: The mismatch negativity (MMN), a well established event related potential (ERP) index of auditory cortex function, was assessed in 12 hallucinating patients (HP), 12 non-hallucinating patients (NP) and 12 healthy controls (HC). The primary endpoints, MMN amplitudes and latencies recorded from anterior and posterior scalp regions, were measured in response to non-phonetic and phonetic sounds. RESULTS: While schizophrenia patients as a whole differed from HCs, no significant between-group differences were observed when patients were divided into hallucinated and non-hallucinated subgroups but, compared to NPs and HCs, whose MMN amplitudes were greatest in response to across phoneme change at frontal but not temporal sites, MMN amplitudes in HPs at frontal sites were not significantly different to any of the presented stimuli, while temporal MMNs in HPs were maximally sensitive to phonetic change. SIGNIFICANCE: These findings demonstrate that auditory verbal hallucinations are associated with impaired pre-attentive processing of speech in fronto-temporal networks, which may involve defective attribution of significance that is sensitive to resource limitations. Overall, this research suggests that MMN may be a useful non invasive tool for probing relationships between hallucinatory and neural states within schizophrenia and the manner in which auditory processing is altered in these afflicted patients. PMID- 18511140 TI - Molecular typing of industrial strains of Pseudomonas spp. isolated from milk and genetical and biochemical characterization of an extracellular protease produced by one of them. AB - P. fluorescens is responsible for the highest depredation of milk because of its capacity to synthesize extracellular lipase and protease which hydrolyze milk fat and proteins. Several P. fluorescens synthesize an extracellular caseinolytic metalloprotease, called AprX. It is important to rapidly detect the presence of a contamination of raw milk by a strain, especially a P. fluorescens strain, having a high potential of depredation. If standard plate count procedures are often employed, they are time consuming and do not permit to rapidly evaluate the potential of depredation. An alternative method consists to search the aprX gene, but such a method remains of low sensitivity and does not allow evaluating the real potential of depredation of the contaminant. After a milk depredation event, three strains of Pseudomonas spp. (F, 2312 and 2313) have been isolated from a dairy plant. Using molecular and phenotypic approaches, these strains were identified as P. fluorescens strains. Their respective extracellular caseinolytic potential was characterized as well as that of several collection strains of P. fluorescens. It appeared that these strains secreted one protease of about 45 kDa, that their extracellular caseinolytic potential was highly variable for one strain to another and that the one of strain F was the highest. The protease secreted by the strain F was purified and its N-terminal sequence established. It shared 100% identity with the domain 14-34 of extracellular alkaline endoprotease sequences which are called AprX for some of them. Its gene was sequenced as well as that of two collection strains of P. fluorescens having a significant lower extracellular caseinolytic potential. The genomic environment of the aprX gene as well as its expression during the strain growth was investigated. It appears that the difference of extracellular caseinolytic potential which has been observed between the three strains does not mainly result from the AprX sequence/structure but it might rather result from the aprX level of expression. PMID- 18511141 TI - Tunisian Salvia officinalis L. and Schinus molle L. essential oils: their chemical compositions and their preservative effects against Salmonella inoculated in minced beef meat. AB - The essential oils (EOs) extracted from the aerial parts of cultivated Salvia officinalis L. and the berries of Schinus molle L. were analysed by gas chromatography-mass spectrometry (GC-MS) and 68 and 67 constituents were identified, respectively. The major constituents were 1,8-cineole (33.27%), beta thujone (18.40%), alpha-thujone (13.45%), borneol (7.39%) in S. officinalis oil and alpha-phellandrene (35.86%), beta-phellandrene (29.3%), beta-pinene (15.68%), p-cymene (5.43%) and alpha-pinene (5.22%) in S. molle oil. In its second part, the present study was conducted to evaluate the in vitro antimicrobial activity of both studied EOs. For this purpose, paper disc-diffusion method and broth microdilution test were used. The disc-diffusion method showed significant zone of lysis against all the pathogens studied (gram-negative and gram-positive bacteria, yeast). These activities remained stable after six months, and decreased approximately by 20% after one year of storage of the EOs at 4 to 7 degrees C. On comparing the efficiency of both EOs, S. officinalis EO exhibited higher antibacterial activity against the majority of strains and especially against Candida albicans (two fold more active according to the inhibition zones values). The minimal inhibitory concentrations (MICs) were reported between 4.5 mg/ml and 72 mg/ml on nutrient broth. The particular chemotype of each EO may be involved in its specific antimicrobial behaviour. Furthermore, the inhibitory effect of these EOs were evaluated against two foodborne pathogens belonging to Salmonella genus, experimentally inoculated (10(3) CFU/g) in minced beef meat, which was mixed with different concentrations of the EO and stored at 4 to 7 degrees C for 15 days. Although the antibacterial activities of both EOs in minced beef meat were clearly evident, their addition had notable effects on the flavour and taste of the meat at concentrations more than 2% for S. molle and 1.5% for S. officinalis. One solution to the above-mentioned problem may be the use of combinations of different food preservation systems. In this context, each of the EOs has been used along with low water activity (addition of NaCl) in addition to low refrigeration temperatures. Results on the Salmonella growth showed that some combinations could be recommended to eliminate germs from minced raw beef. By using this method, a stable and, from a microbiological point of view, safe meat can be produced without substantial loss in sensory quality. Results obtained herein, may suggest that the EOs of S. officinalis and S. molle possess antimicrobial activity, and therefore, they can be used in biotechnological fields as natural preservative ingredients in food and/or pharmaceutical industry. PMID- 18511142 TI - Bifurcational and dynamical analysis of a continuous biofilm reactor. AB - A dynamical model of a continuous biofilm reactor is presented. The reactor consists of a three-phase internal loop airlift operated continuously with respect to the liquid and gaseous phases, and batchwise with respect to the immobilized cells. The model has been applied to the conversion of phenol by means of immobilized cells of Pseudomonas sp. OX1 whose metabolic activity was previously characterized (Viggiani, A., Olivieri, G., Siani, L., Di Donato, A., Marzocchella, A., Salatino, P., Barbieri, P., Galli, E., 2006. An airlift biofilm reactor for the biodegradation of phenol by Pseudomonas stutzeri OX1. Journal of Biotechnology 123, 464-477). The model embodies the key processes relevant to the reactor performance, with a particular emphasis on the role of biofilm detachment promoted by the fluidized state. Results indicate that a finite loading of free cells establishes even under operating conditions that would promote wash out of the suspended biophase. The co-operative/competitive effects of free cells and immobilized biofilm result in rich bifurcational patterns of the steady state solutions of the governing equations, which have been investigated in the phase plane of the process parameters. Direct simulation under selected operating conditions confirms the importance of the dynamical equilibrium establishing between the immobilized and the suspended biophase and highlights the effect of the initial value of the biofilm loading on the dynamical pattern. PMID- 18511143 TI - Comparative genomics of serotype Asia 1 foot-and-mouth disease virus isolates from India sampled over the last two decades. AB - This study deals with a comparative analysis of complete genome sequences of twenty-one serotype Asia 1 foot-and-mouth disease (FMD) field viruses isolated over a period of two decades from India, two vaccine strains and seven exotic sequences. The Indian viruses could be grouped in to three distinct lineages at the entire coding region, evolving independently probably under differential selection pressure as evident from the lineage-specific signatures identified. This comparison revealed 80% of amino acids at the polyprotein region to be invariant. Twenty-one residues in L, 3A and P1 region were identified to be under positive selection of which some are antigenically critical. Analysis at functionally crucial motifs, receptor contact residues, polyprotein cleavage sites and at putative T-cell epitopes expands the knowledge beyond other serotypes. Antigenic site II in betaB-betaC loop of VP2 was highly unstable suggesting its exposure to extreme immune pressure. A single cross-over at the L proteinase region in an isolate from buffalo, also featuring an extensive deletion at the 5' untranslated region (UTR), reflects the role of intraserotypic genetic recombination in natural evolution. The likely biological relevance of deletions/insertions observed at UTRs, VP1 and 3A could not be deduced. Altogether, a substantial amount of data raised on full length genomes of type Asia 1 virus adds value to the FMD virus genomics. PMID- 18511144 TI - Three heterologous proteins simultaneously expressed from a chimeric potyvirus: infectivity, stability and the correlation of genome and virion lengths. AB - Three heterologous proteins were simultaneously expressed from a chimeric potyvirus Potato virus A (PVA) in Nicotiana benthamiana. The genes for green fluorescent protein of Aequoria victoriae ("G"; 714 nucleotides, nt), luciferase of Renilla reniformis ("L", 933 nt) and beta-glucuronidase of Escherichia coli ("U", 1806 nt) were inserted into the engineered cloning sites at the N-terminus of the P1 domain, the junction of P1 and helper component protein (HC-Pro), and the junction of the viral replicase (NIb) and coat protein (CP), respectively, in an infectious PVA cDNA. The proteins were expressed as part of the viral polyprotein and subsequently released by cleavage at the flanking proteolytic cleavage sites by P1 (one site) or the NIa-Pro proteinase (other sites). The engineered viral genome (pGLU, 13311 nt) was 39.2% larger than wild-type PVA (9565 nt) and infected plants of N. benthamiana systemically. pGLU was stable and expressed all three heterologous proteins, also following the second infection cycle initiated by sap-inoculation of new plants with the progeny viruses. The gene for GUS showed some inherent instabilities, as also reported in other studies. Accumulation of pGLU in infected leaves was lower by a magnitude as compared to the vector viruses pG0U and p0LU used to express two heterologous proteins. Hence, pGLU may have reached the maximum genome size that can still function and complete the PVA infection cycle. Examination of virions by electron microscopy indicated that the virion lengths of PVA chimera with various numbers of inserts were directly proportional to their genome lengths. PMID- 18511145 TI - High loading efficiency and tunable release of plasmid DNA encapsulated in submicron particles fabricated from PLGA conjugated with poly-L-lysine. AB - Poly(lactic-co-glycolic acid) (PLGA) particles have been widely explored as vehicles for delivery of plasmid DNA to mammalian cells both in vitro and in vivo. Achieving high incorporation efficiencies and control over release kinetics are significant challenges in encapsulating hydrophilic molecules such as DNA within submicron particles fabricated from PLGA. This study explored two modifications in the preparation of submicron particles to specifically address these challenges. Firstly, we compared homogenization and sonication as energy sources for emulsification. It was demonstrated that particles prepared with homogenization resulted in higher encapsulation efficiency and a linear release profile of DNA as compared to particles prepared with sonication, which exhibited lower encapsulation efficiency and a burst release. Also investigated was conjugation of poly-L-lysine to PLGA (PLGA-PLL) to create an electrostatically favorable interaction between the carrier material and the DNA. Particles fabricated with high weight percentages of PLGA-PLL/PLGA resulted in remarkably increased loading (>90%). Additionally, the release profile could be dictated by the quantity of PLGA-PLL incorporated into the particles. Particles incubated in vitro on COS-7 cells were able to transfect cells. These results demonstrated that DNA encapsulation and release were modulated by the method of fabrication. PMID- 18511146 TI - Skipping meals and alcohol consumption. The regulation of energy intake and expenditure among weight loss participants. AB - Research suggests that specific eating patterns (e.g., eating breakfast) may be related to favorable weight status. This investigation examined the relationship between eating patterns (i.e., skipping meals; consuming alcohol) and weight loss treatment outcomes (weight loss, energy intake, energy expenditure, and duration of exercise). Fifty-four overweight or obese adults (BMI> or =27 kg/m(2)) participated in a self-help or therapist-assisted weight loss program. Daily energy intake from breakfast, lunch, dinner, and alcoholic beverages, total daily energy intake, total daily energy expenditure, physical activity, and weekly weight loss were assessed. On days that breakfast or dinner was skipped, or alcoholic beverages were not consumed, less total daily energy was consumed compared to days that breakfast, dinner, or alcoholic beverages were consumed. On days that breakfast or alcohol was consumed, daily energy expenditure (breakfast only) and duration of exercise were higher compared to days that breakfast or alcohol was not consumed. Individuals who skipped dinner or lunch more often had lower energy expenditure and exercise duration than individuals who skipped dinner or lunch less often. Individuals who consumed alcohol more often had high daily energy expenditure than individuals who consumed alcohol less often. Skipping meals or consuming alcoholic beverages was not associated with weekly weight loss. In this investigation, weight loss program participants may have compensated for excess energy intake from alcoholic beverages and meals with greater daily energy expenditure and longer exercise duration. PMID- 18511147 TI - Clindamycin and metronidazole as independent risk factors for nosocomial acquisition of multidrug-resistant Pseudomonas aeruginosa. PMID- 18511148 TI - Distribution of the antiseptic-resistance gene qacEDelta1 in 283 clinical isolates of Gram-negative bacteria in China. PMID- 18511149 TI - Optimising peripheral vascular catheter care offers the greatest potential for prevention of vascular-device-related infections. PMID- 18511150 TI - VITEK 2 failure in screening Hafnia alvei inducible beta-lactam resistance. PMID- 18511151 TI - Different ribotypes in community-acquired Clostridium difficile. PMID- 18511152 TI - Web-based reporting of the results of the 2006 four country prevalence survey of healthcare associated infections. AB - A web-base reporting system was developed in order to feed back the results of the 2006 prevalence survey of healthcare-associated infections in a timely manner to all participating hospitals in England, Wales and Northern Ireland. The database accommodated approximately 75 000 records from over 250 hospitals. The reporting system was hosted on the National Health Service intranet, accessible via secure login. Users were able to access their individual Trust data via a series of predefined reports and an export facility was included to facilitate additional analysis. The reporting system was made available to participating hospitals within 12 months of completion of the survey. From the results of a user satisfaction survey, end-users responded positively to receiving feedback in this format. It serves as a useful model for the feedback of results for future prevalence surveys. PMID- 18511153 TI - Reducing blood-culture contamination rates by the use of a 2% chlorhexidine solution applicator in acute admission units. PMID- 18511154 TI - A pseudo-outbreak of Fusarium solani in an intensive care unit associated with bronchoscopy. PMID- 18511155 TI - Luminol-based forensic detection of latent blood; an approach to rapid wide-area screening combined with Glo-Germ oil simulant studies. PMID- 18511156 TI - Nosocomial outbreak of two strains of Burkholderia cepacia caused by contaminated heparin. PMID- 18511157 TI - Analogs of JHU75528, a PET ligand for imaging of cerebral cannabinoid receptors (CB1): development of ligands with optimized lipophilicity and binding affinity. AB - Cyano analogs of Rimonabant with high binding affinity for the cerebral cannabinoid receptor (CB1) and with optimized lipophilicity have been synthesized as potential positron emission tomography (PET) ligands. The best ligands of the series are optimal targets for the future radiolabeling with PET isotopes and in vivo evaluation as radioligands with enhanced properties for PET imaging of CB1 receptors in human subjects. Extracellular electrophysiological recordings in rodent brain slices demonstrated that JHU75528, 4, the lead compound of the new series, has functional CB antagonist properties that are consistent with its structural relationship to Rimonabant. Molecular modeling analysis revealed an important role of the binding of the cyano group with the CB1 binding pocket. PMID- 18511158 TI - Reduced dose pre-exposure primary and booster intradermal rabies vaccination with a purified chick embryo cell vaccine (PCECV) is immunogenic and safe in adults. AB - Pre-exposure vaccination of persons at risk with intradermally administered reduced dose cell culture rabies vaccines remains controversial in low-enzootic countries. In a prospective clinical trial of adult volunteers (N=25), we studied the immune response to purified chick embryo cell vaccine (PCECV) administered intradermally at a reduced dose (0.1 mL) in a three-dose schedule (0, 7 and 21 days). In 10 subjects, immunogenicity of intradermally administered one-dose booster vaccination with 0.1 mL PCECV was investigated. All participants were seroconverted 3 weeks after primary and 1 week after booster vaccination (antibody titre > or = 0.5 EU/mL, measured by enzyme linked immunosorbent assay). Local adverse events such as erythema and swelling were moderate and transitory. The intradermal vaccination route offers an efficacious and cost-reducing strategy to increase the accessibility of cell culture rabies vaccines. PMID- 18511159 TI - Development of a universal influenza A vaccine based on the M2e peptide fused to the papaya mosaic virus (PapMV) vaccine platform. AB - With the emergence of highly virulent influenza viruses and the consequent risk of pandemics, new approaches to designing universal influenza vaccines are urgently needed. In this report, we demonstrate the potential of using a papaya mosaic virus (PapMV) platform carrying the universal M2e influenza epitope (PapMV CP-M2e) as a candidate flu vaccine. We show that PapMV-CP-M2e virus-like particles (VLPs) can induce production in mice of anti-M2e antibodies that can recognize influenza-infected cells. PapMV-CP-M2e discs made of 20 coat protein (CP) subunits were shown to be poorly immunogenic compared to PapMV-CP-M2e VLPs composed of several hundred CP subunits. We also show that addition of either alum or PapMV-CP VLPs as adjuvant dramatically increased the immunogenicity of PapMV-CP-M2e-containing vaccine, and led to 100% protection against a challenge of 4LD(50) with the WSN/33 strain. These results show, for the first time, the potential of a recombinant plant virus protein to serve as both peptide delivery system and adjuvant in the crucial field of influenza vaccine development. PMID- 18511160 TI - Poly(propyleneimine) dendrimer and dendrosome mediated genetic immunization against hepatitis B. AB - The purpose of the present research work is to explore the potential of dendrosomes in genetic immunization against hepatitis B. Plasmid DNA encoding pRc/CMV-HBs[S] (5.6 kb), encoding the small region of the hepatitis B surface antigen, was complexed with 5th generation poly(propyleneimine) dendrimer (PPI) in different ratios. Transfection of CHO cells revealed that a ratio of 1:50 for pDNA:PPI was optimum for transfection. Results of cytotoxicity studies showed that the toxicity of PPI-DNA complex was significantly (p<0.05) higher for PPI 75 and PPI 100 as compared to the other PPI-DNA complexes. PPI 50 was employed for preparation of dendrosomes by reverse phase evaporation method. The dendrosomal formulation DF3 was found to possess optimum vesicle size, zeta potential and entrapment efficiency. In vitro production of HBsAg in CHO cells showed that DF3 possess maximum transfection efficiency. In vivo immunization studies were carried out by giving a single intramuscular injection of 10 microg of plasmid DNA (pDNA) or its dendrimeric or dendrosomal formulation to female Balb/c mice, followed by estimation of total IgG, IgG(1), IgG(2a), IgG(2b), biweekly. DF3 was found to elicit maximum immune response in terms of total IgG and its subclasses under study as compared to PPI 50 and pDNA at all time points. Animals immunized with DF3 developed very high cytokine level. Higher level of IFN-gamma suggests that the immune response was strictly Th1 mediated. Our observations clearly prove the superiority of dendrosomes over PPI-DNA complex and pDNA for genetic immunization against hepatitis B. PMID- 18511161 TI - Oblique use of a trephine bur for the harvesting of tibial bone grafts. PMID- 18511162 TI - Management of the giant frontal sinus--a simple method to improve cosmesis. AB - Frontal bossing can be caused by many craniofacial syndromes, however an enlarged frontal sinus is a rare cause. Because of this the management of this condition is variable. One option is to remove the anterior wall of the frontal sinus and dividing this into segments which are then fixed in the desired position using miniplates.This however may prove difficult in thin bone and in these cases the periosteum can be left intact to splint the fragments which are then repositioned using resorbable sutures. PMID- 18511163 TI - Thalassemia and hypercoagulability. AB - Thalassemia is a congenital hemolytic disease caused by defective globin synthesis resulting in decreased quantity of globin chains. Although the life expectancy of beta-thalassemia patients has markedly improved over the last few years, patients still suffer from many complications of this congenital disease. The presence of a high incidence of thromboembolic events, mainly in beta thalassemia intermedia, has led to the identification of a hypercoagulable state in these patients. In this paper, we review the molecular and cellular mechanisms leading to hypercoagulability in beta-thalassemia, with a special focus on thalassemia intermedia being the group with the highest incidence of thrombotic events as compared to other types of thalassemias. We also discuss the recommendations for thrombosis prophylaxis in these patients. PMID- 18511164 TI - Recovery of kidney function after acute kidney injury in the elderly: a systematic review and meta-analysis. AB - BACKGROUND: The prevalence of elderly individuals continues to increase over time, as does the incidence of acute kidney injury (AKI). However, it is not known whether age is an important prognostic predictor for renal recovery after an episode of AKI. STUDY DESIGN: Systematic review of MEDLINE and EMBASE databases and meta-analysis of pooled data using random-effect models. SETTING & POPULATION: Adults with AKI, not including kidney transplant recipients. SELECTION CRITERIA FOR STUDIES: Studies published in English between 2000 and 2007 were eligible for this analysis if they met the following inclusion criteria: (1) clear definition of AKI and recovery of kidney function, (2) assessment of kidney function recovery as the primary or secondary outcome, and (3) participant age reported. We contacted the investigators of studies and requested data for recovery of kidney function by patient age. PREDICTOR: Patient age of 65 years and older and younger than 65 years. OUTCOMES: Recovery of kidney function defined as independence from dialysis therapy, decrease in serum creatinine level to less than a defined threshold, or return to baseline kidney function. RESULTS: We obtained data for recovery of kidney function by age from 17 studies of patients with AKI. Overall, 31.3% of surviving elderly patients did not recover kidney function compared with 26% of younger patients (pooled relative risk, 1.28, 95% confidence interval, 1.06 to 1.55; P < 0.05). The increased risk of nonrecovery in the elderly remained greater in several subgroups examined through sensitivity analyses, including those stratified by type of dialysis support, time of assessment of recovery (short versus long term), and definition of renal recovery. LIMITATIONS: There was significant heterogeneity among studies with respect to comorbid factors, definition of AKI, and study design. CONCLUSIONS: There is impaired recovery of kidney function after AKI in aged individuals. Future studies should be cognizant of "age" as a potential effect modifier in the prognosis after AKI, and clinical trials should focus on improving outcomes in the elderly cohort. PMID- 18511165 TI - Cognitive-behavioral therapy for sleep disturbance in patients undergoing peritoneal dialysis: a pilot randomized controlled trial. AB - BACKGROUND: Greater than 50% of dialysis patients experience sleep disturbances. Cognitive-behavioral therapy (CBT) is effective for treating chronic insomnia, but its effectiveness has never been reported in peritoneal dialysis (PD) patients and its association with cytokines is unknown. We investigated the effectiveness of CBT in PD patients by assessing changes in sleep quality and inflammatory cytokines. STUDY DESIGN: Randomized control study with parallel group design. SETTING & PARTICIPANTS: 24 PD patients with insomnia in a tertiary medical center without active medical and psychiatric illness were enrolled. INTERVENTION: The intervention group (N = 13) received CBT from a psychiatrist for 4 weeks and sleep hygiene education, whereas the control group (N = 11) received only sleep hygiene education. OUTCOMES & MEASUREMENTS: Primary outcomes were changes in the Pittsburgh Sleep Quality Index and Fatigue Severity Scale scores, and secondary outcomes were changes in serum interleukin 6 (IL-6), IL 1beta, IL-18, and tumor necrosis factor alpha levels during the 4-week trial. RESULTS: Median percentages of change in global Pittsburgh Sleep Quality Index scores were -14.3 (interquartile range, -35.7 to - 6.3) and -1.7 (interquartile range, -7.6 to 7.8) in the intervention and control groups, respectively (P = 0.3). Median percentages of change in global Fatigue Severity Scale scores were 12.1 (interquartile range, -59.8 to -1.5) and -10.5 (interquartile range, -14.3 to 30.4) in the intervention and control groups, respectively (P = 0.04). Serum IL-1beta level decreased in the intervention group, but increased in the control group (P = 0.04). There were no significant differences in changes in other cytokines. LIMITATIONS: This study had a small number of participants and short observation period, and some participants concurrently used hypnotics. CONCLUSIONS: CBT may be effective for improving the quality of sleep and decreasing fatigue and inflammatory cytokine levels. CBT can be an effective nonpharmacological therapy for PD patients with sleep disturbances. PMID- 18511166 TI - Association between a self-rated health question and mortality in young and old dialysis patients: a cohort study. AB - BACKGROUND: Self-rated health (SRH) has been shown to predict mortality in large community-based studies; however, large clinical-based studies of this topic are rare. We assessed whether an SRH item predicts mortality in a large sample of incident dialysis patients beyond sociodemographic, disease, and clinical measures and possible age interaction. STUDY DESIGN: Prospective cohort study. SETTING & PARTICIPANTS: 1,443 predominantly white patients from 38 dialysis centers in The Netherlands participating in the Netherlands Cooperative Study on the Adequacy of Dialysis-2 between 1997 and 2004. PREDICTOR: SRH score completed at 3 months after the start of dialysis therapy (baseline). OUTCOMES & MEASUREMENTS: Cox proportional hazards model estimated the association between SRH and all-cause mortality. Interaction of SRH with age (<65 and >/=65 years) was examined in an additive model. RESULTS: Mean age of patients was 59.6 +/- 14.8 years, with 61% men and 69% married/living together. Mean follow-up was 2.7 +/- 1.8 years. Deaths per SRH group in the multivariate analyses sample: excellent/very good (9 of 63 patients; 14.3%), good (148 of 473 patients; 31.3%), fair (194 of 508 patients; 38.2%), and poor (45 of 71 patients; 63.4%). Patients with poor, fair, or good health ratings had a greater mortality risk than those with excellent/very good health ratings (adjusted hazard ratio [HR(adj)], 3.56; 95% confidence interval [CI], 1.71 to 7.42; HR(adj), 2.09; 95% CI, 1.06 to 4.12; HR(adj), 1.87; 95% CI, 0.95 to 3.70, respectively) independent of a range of risk factors. No age interaction with SRH was found. LIMITATIONS: Although the SRH mortality association remained strong despite extensive adjustments, unknown residual confounding could still exist. CONCLUSION: SRH is an independent predictor of mortality in incident dialysis patients. Patients with poor SRH in both age strata had a significantly increased risk of mortality even after controlling for demographic and clinical confounders. Patient self-assessment of health can be an invaluable and economical complement to clinical measures in risk assessment. PMID- 18511167 TI - Intravascular hemolysis and acute renal failure after mitral and aortic valve repair. PMID- 18511168 TI - Waist-to-hip ratio, body mass index, and subsequent kidney disease and death. AB - BACKGROUND: Chronic kidney disease (CKD) and obesity are important public health concerns. We examined the association between anthropomorphic measures and incident CKD and mortality. STUDY DESIGN: Cohort study. SETTING & PARTICIPANTS: Individual patient data pooled from the Atherosclerosis Risk in Communities Study and the Cardiovascular Health Study. PREDICTORS: Waist-to-hip ratio (WHR), body mass index (BMI). OUTCOMES & MEASUREMENTS: Incident CKD defined as serum creatinine level increase greater than 0.4 mg/dL with baseline creatinine level of 1.4 mg/dL or less in men and 1.2 mg/dL or less in women and final creatinine level greater than these levels, and, in separate analyses, estimated glomerular filtration rate (eGFR) decrease of 15 mL/min/1.73 m(2) or greater with baseline eGFR of 60 mL/min/1.73 m(2) or greater and final eGFR less than 60 mL/min/1.73 m(2). Multivariable logistic regression to determine the association between WHR, BMI, and outcomes. Cox models to evaluate a secondary composite outcome of all cause mortality and incident CKD. RESULTS: Of 13,324 individuals, mean WHR was 0.96 in men and 0.89 in women and mean BMI was 27.2 kg/m(2) in both men and women. During 9.3 years, 300 patients (2.3%) in creatinine-based models and 710 patients (5.5%) in eGFR-based models developed CKD. In creatinine-based models, each SD increase in WHR was associated with increased risk of incident CKD (odds ratio, 1.22; 95% confidence interval [CI], 1.05 to 1.43) and the composite outcome (hazard ratio, 1.12; 95% CI, 1.06 to 1.18), whereas each SD increase in BMI was not associated with CKD (odds ratio, 1.05; 95% CI, 0.93 to 1.20) and appeared protective for the composite outcome (hazard ratio, 0.94; 95% CI, 0.90 to 0.99). Results of eGFR-based models were similar. LIMITATIONS: Single measures of creatinine, no albuminuria data. CONCLUSIONS: WHR, but not BMI, is associated with incident CKD and mortality. Assessment of CKD risk should use WHR rather than BMI as an anthropomorphic measure of obesity. PMID- 18511169 TI - Kotomolide A arrests cell cycle progression and induces apoptosis through the induction of ATM/p53 and the initiation of mitochondrial system in human non small cell lung cancer A549 cells. AB - This study first investigates the anticancer effect of kotomolide A (KTA) in human non-small cell lung cancer cells, A549. KTA has exhibited effective cell growth inhibition by inducing cancer cells to undergo G2/M phase arrest and apoptosis. Blockade of cell cycle was associated with increased the activation of ataxia telangiectasia-mutated (ATM). Activation of ATM by KTA phosphorylated p53 at Serine15, resulting in increased stability of p53 by decreasing p53 and murine double minute-2 (MDM2) interaction. In addition, KTA-mediated G2/M phase arrest also was associated with the decrease in the amounts of cyclinB1, cyclinA, Cdc2 and Cdc25C and increase in the phosphorylation of Chk2, Cdc25C and Cdc2. Specific ATM inhibitor, caffeine, significantly decreased KTA-mediated G2/M arrest by inhibiting the phosphorylation of p53 (Serine15) and Chk2. KTA treatment triggered the mitochondrial apoptotic pathway indicated by a change in Bax/Bcl-2 ratios, resulting in mitochondrial membrane potential loss and caspase-9 activation. Taken together, these results suggest a critical role for ATM and p53 in KTA-induced G2/M arrest and apoptosis of human non-small cell lung cancer cells. PMID- 18511170 TI - Toxicity and gastric tolerance of essential oils from Cymbopogon citratus, Ocimum gratissimum and Ocimum basilicum in Wistar rats. AB - Oils of Cymbopogon citratus, Ocimum gratissimum and Ocimum basilicum are widely used for their medicinal properties, and as food flavours and perfumes. Recently in a study in West Africa, these oils have been recommended to combat Fusarium verticillioides and subsequent fumonisin contamination in stored maize, but their toxicological profile was not investigated. The current study was undertaken to provide data on acute and subacute toxicity as well as on gastric tolerance of these oils in rat. For this purpose, the oils were given by gavage to Wistar rats for 14 consecutive days. The animals were observed daily for their general behaviour and survival, and their visceral organs such as stomach and liver were taken after sacrifice for histological analyses. A dose-dependent effect of the tested oils was observed during the study. Applied at doses generally higher than 1500 mg/kg body weight, the oils caused significant functional damages to stomach and liver of rat. Unlike the other oils, administration of O. gratissimum oil did not result in adverse effects in rat liver at the tested doses. The no observed adverse effect level (NOAEL) of the tested oils has been established. The three tested oils can be considered as safe to human when applied on stored maize at recommended concentrations. PMID- 18511171 TI - Fluoride at non-toxic dose affects odontoblast gene expression in vitro. AB - Elevated fluoride intake may lead to local tissue disturbances, known as fluorosis. Towards an understanding of this effect, fluoride-induced molecular responses were analyzed in MO6-G3 cultured odontoblasts cells. NaF at 1mM changed expression of genes implicated in tissue formation and growth, without affecting cell proliferation or inducing stress factor RNAs. Up to 1mM NaF, DNA accumulation was not inhibited, whereas at 3mM, cells detached from their support and did not proliferate. Intracellular structures, characterized by EM, were normal up to 1mM, but at 3mM, necrotic features were evident. No sign of apoptotic transformation appeared at any NaF concentration. Fluoride-sensitive genes were identified by microarray analysis; expression levels of selected RNAs were determined by conventional and real-time RT-PCR. At 1mM fluoride, RNAs encoding the extracellular matrix proteins asporin and fibromodulin, and the cell membrane associated proteins periostin and IMT2A were 10-fold reduced. RNA coding for signaling factor TNF-receptor 9 was diminished to one-third, whereas that for the chemokine Scya-5 was enhanced 2.5-fold. These RNAs are present in vivo in tooth forming cells. This was demonstrated by in situ hybridization and RT-PCR on RNA from dissected tissue samples; for the presence and functioning of fibromodulin in dentin matrix, a more comprehensive study has earlier been performed by others [Goldberg, M., Septier, D., Oldberg, A., Young, M.F., Ameye, L.G., 2006. Fibromodulin deficient mice display impaired collagen fibrillogenesis in predentin as well as altered dentin mineralization and enamel formation. J. Histochem. Cytochem. 54, 525-537]. Expression of most other RNA species, in particular of stress factor coding RNAs, was not altered. It was concluded that fluoride could influence the transcription pattern without inducing cell stress or apoptosis. In odontoblasts in vivo, aberrant expression of these fluoride sensitive genes may impair the formation of the extracellular matrix and influence cell communication, with the possible consequence of fluorotic patterns of normal and deviant dentin. PMID- 18511172 TI - Exploiting endogenous anti-apoptotic proteins for novel therapeutic strategies in cerebral ischemia. AB - The acute neuronal degeneration in the ischemic core upon stroke is followed by a second wave of cell demise in the ischemic penumbra and neuroanatomically connected sites. This temporally delayed deleterious event of programmed cell death ('secondary degeneration') often exceeds the initial damage of stroke and, thus, contributes pivotally to significant losses in neurological functions. In fact, it is the injured neurons in these regions around the ischemic core zone that neuropharmacological prevention is targeting to preserve. Clinical and pre clinical studies have focussed on neuroprotective interventions with caspase inhibitors, but it remains ambiguous whether diminishing or even silencing these aspartate-specific cysteine proteases are in sum beneficial for the clinical outcome. It is often ignored that caspase inhibitors are able to antagonize calpain and cathepsins, thereby protecting the cytoskeleton from damage. Moreover, there is a point of no return, beyond which interfering with caspases cannot rescue the cell, but spoil the obligate and necessary suicide program such that the cellular environment suffers from by-products of necrosis and secondary inflammation. Here we discuss novel alternative strategies to abrogate the death cascade at the level of the genomic response (transcription factors, NF-kappaB, CREB, ICER, HIF), of mitochondrial effectors (cytochrome c, Bcl-2, Smac/DIABLO, HtrA2), and of inhibitor of apoptosis proteins (IAPs). IAPs are the only known endogenous proteins that inhibit specifically and with high affinity the activity of both initiator and effector caspases. Based on compelling biochemical evidence, we argue that patronizing the neuronal endogenous anti-apoptotic machinery could be superior to the pharmacological inhibition of caspases at various levels, with regard to specificity, side effects, and the 'therapeutic window of opportunity'. PMID- 18511173 TI - Neurobiology of cognitive aging: insights from imaging genetics. AB - Over the last several years, neuroscientists have been increasingly using neuroimaging techniques to unravel the neurobiology underlying cognitive aging, and in more recent years to explore the role of genes on the variability of the aging process. One of the primary goals of this research is to identify proteins involved in cognitive aging with the hope that this would facilitate the development of novel treatments to combat cognitive impairment. Further, it is likely with early identification of susceptible individuals, early intervention through life-style changes and other methods could increase an individual's resilience to the effects of aging. PMID- 18511174 TI - Immunohistochemical localization of alpha-tocopherol transfer protein and lipoperoxidation products in human first-trimester and term placenta. AB - OBJECTIVE: Pregnancy is described as a state of oxidative stress arising from the high metabolic turnover taking place during feto-placental development and little is known about the balance of oxidation and antioxidation in early human pregnancy. The aim of this study was to analyze placental expression of alpha tocopherol transfer protein (alpha-TTP) as the major transport protein for the antioxidant alpha-tocopherol as well as the placental expression of two lipoperoxidation products, malondialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE) in early first-trimester and term human placenta. STUDY DESIGN: Placental tissue was obtained from 10 pregnancy interruptions at 6-8 weeks gestational age and 10 samples were obtained from term pregnancies after routine cesarean section. The placental expression of alpha-TTP, MDA and HNE has been investigated with immunohistochemistry by the use of specific human alpha-TTP, MDA and HNE antibodies. RESULTS: While MDA and HNE showed similar expression in first trimester and term placenta, alpha-TTP expression was less in first-trimester syncytiotrophoblast as compared to term. In first-trimester specimen, alpha-TTP showed major expression in extravillous trophoblast. In amniotic epithelial cells, a rising tendency in all three parameters investigated from immature to mature cells could be documented. No direct correlation between alpha-TTP, MDA and HNE expression was detected. CONCLUSIONS: Our study shows the presence of alpha-TTP not only in term, but in first-trimester extravillous trophoblast, syncytiotrophoblast and amniotic epithelium. Furthermore, lipoperoxidation products MDA and HNE are also present in first-trimester and term placenta, documenting the presence of oxidative processes in the placenta from early on. It therefore seems possible that scavenging of reactive oxygen species (ROS) by alpha-tocopherol is already required in first-trimester human pregnancy, but the missing correlation to MDA and HNE expression leads to the speculation that alpha TTP and its ligand alpha-tocopherol have functions beyond the antioxidative capacity of alpha-tocopherol in early pregnancy. PMID- 18511175 TI - The efficacy of alum-containing ferrous thermal water in the management of chronic inflammatory gynaecological disorders--a randomized controlled study. AB - OBJECTIVES: Treatment of gynaecological disorders is a frequent, but only barely substantiated application of balneotherapy. This study investigated potential differences between the clinical symptoms, pelvic blood flow and specific laboratory parameters of patients undergoing balneotherapy with two different types of immersion: alum-containing and tap water. STUDY DESIGN: The study population comprised 40 patients (mean age: 39.4 years), randomized into two groups. All subjects took 20 min baths in 38 degrees C water every other day, for 10 occasions altogether. Study parameters were: pain relief, reduction in tissue growth, hormone levels, psychic status, and pelvic blood flow. RESULTS: Thermal water improved the clinical parameters of both groups significantly. In comparison with tap water, treatment with alum-containing water accomplished significantly greater progress, as reflected by the relief of pain elicited by handling the uterus and improvement of psychic status. Laboratory parameters (FSH, LH, prolactin, oestradiol and beta-endorphin serum levels) and the Doppler index did not change in either group. CONCLUSIONS: As demonstrated by our results, 3-week balneotherapy is a potentially useful adjunct for the management of chronic pelvic inflammatory disease, but further, long-term studies are notwithstanding necessary. PMID- 18511176 TI - Editorial comment on: surveillance study in Europe and Brazil on clinical aspects and Antimicrobial Resistance Epidemiology in Females with Cystitis (ARESC): implications for empiric therapy. PMID- 18511177 TI - The comparability of models for predicting the risk of a positive prostate biopsy with prostate-specific antigen alone: a systematic review. AB - CONTEXT: The sensitivity and specificity profile of measuring levels of prostate specific antigen (PSA) to select men for prostate biopsy is not optimal. This has prompted the construction of nomograms and artificial neural networks (ANNs) to increase the performance of PSA measurements. OBJECTIVE: A systematic review of nomograms and ANNs designed to predict the risk of a positive prostate biopsy for cancer was conducted in order to determine their value versus measuring PSA levels alone. EVIDENCE ACQUISITION: Medical Literature Analysis and Retrieval System Online (U.S. National Library of Medicine's life science database; MEDLINE) was searched using the terms "nomogram" "artificial neural network" and "prostate cancer" for dates up to and including July 2007 and was supplemented by manual searches of reference lists. Included studies used an assessment tool to examine the risk of a positive prostate biopsy in a man without a known cancer diagnosis. Intramodel comparisons with evaluation of PSA levels alone, and intermodel comparisons of area under the curve (AUC) from receiver operating characteristic (ROC) curves were conducted. Individual case examples were also used for comparisons. EVIDENCE SYNTHESIS: Twenty-three studies examining 36 models were included. With the exception of two studies, all the models had AUC values of 0.70 or greater, with eight reporting an AUC of >/=0.80 and four (all ANNs) reporting an AUC >/=0.85, with variable validation status. Fourteen studies compared the AUC with PSA levels alone: all showed a benefit from using AUCs which varied from 0.02 to 0.26. Of the 16 external validation comparisons, in 13 the AUC was lower in the external population than in the model population. CONCLUSIONS: Nomograms and ANNs produce improvements in AUC over measurement of PSA levels alone, but many lack external validation. Where this is available, the benefits are often diminished, although most remain significantly better than with evaluation of PSA levels alone. In men without additional risk factors, PSA cutoff values alone provide a relatively precise risk estimate, but if additional risk factors are known, PSA values alone are less accurate. PMID- 18511178 TI - Surveillance study in Europe and Brazil on clinical aspects and Antimicrobial Resistance Epidemiology in Females with Cystitis (ARESC): implications for empiric therapy. AB - BACKGROUND: Uncomplicated cystitis in females is among the most frequent infections in community. OBJECTIVE: To determine clinical aspects, epidemiology, and antimicrobial susceptibility of uropathogens. INTERVENTION: Patients were investigated clinically and with urinalysis and urine culture. MEASUREMENTS: This survey started in 2003 and ended in 2006 including 68 centres in nine European countries and in Brazil. Female patients between 18 and 65 yr with symptoms of uncomplicated cystitis were consecutively enrolled and clinically evaluated. Uropathogens were identified and their susceptibility tested for nine antimicrobials. RESULTS AND LIMITATIONS: Clinical data of 4264 eligible patients were analysed. A positive urine culture was found in 74.6%. Within the 3018 pathogens, Escherichia coli (E. coli) was most frequent (76.7%), followed by Enterococcus faecalis (4.0%), Staphylococcus saprophyticus (3.6%), Klebsiella pneumoniae (3.5%), and Proteus mirabilis (3.5%). E. coli showed the highest rate of susceptibility to fosfomycin (98.1%) followed by mecillinam (95.8%), nitrofurantoin (95.2%), and ciprofloxacin (91.8%). The lowest rate was found for ampicillin (45.1%). For the total spectrum the order was fosfomycin (96.4%), mecillinam (95.9%), ciprofloxacin (90.3%), and nitrofurantoin (87.0%). In all countries a susceptibility rate to E. coli above 90% was found only for fosfomycin, mecillinam, and nitrofurantoin. The susceptibility rates varied significantly from country to country (p<0.0001), except for fosfomycin, mecillinam, and nitrofurantoin. CONCLUSIONS: Despite wide cross-country variability of bacterial susceptibility/resistance rates to the other antimicrobials tested, fosfomycin, mecillinam, and nitrofurantoin have preserved their in vitro activity in all countries investigated. They may represent good options for the empiric therapy of female patients with uncomplicated cystitis. PMID- 18511179 TI - Editorial comment on: Hydrogen sulphide is involved in testosterone vascular effect. PMID- 18511180 TI - Hydrogen sulphide is involved in testosterone vascular effect. AB - BACKGROUND: Testosterone (T) induces a rapid relaxation in vascular tissues of different species due to a nongenomic effect of this steroid on vessels. Different mechanisms have been proposed to explain T-induced vasodilatation but the effective mechanism(s) and the mediators involved are still a matter of debate. OBJECTIVES: We have evaluated if H(2)S pathway is involved in T vascular effects. DESIGN AND SETTING: Male Wistar rats were sacrificed and thoracic aorta was rapidly dissected and cleaned from fat and connective tissue. Rings of 2-3 mm length were cut and placed in organ baths filled with oxygenated Krebs solution at 37 degrees C and mounted to isometric force transducers. H(2)S determination was performed on thoracic aortic rings incubated with T or vehicle and in presence of inhibitors. H2S concentration was calculated against a calibration curve of NaHS (3-250 microM). Results were expressed as nmoles/mg protein. MEASUREMENTS: Vascular reactivity was evaluated by using isometric transducers. H(2)S determination was performed by using a cystathionine beta-synthetase (CBS) and cystathionine gamma lyase (CSE) activity assay. CSE and CBS protein levels were assessed by Western blot analysis. Statistical analysis was performed by using two-way ANOVA and unpaired Student's t-test where appropriate. RESULTS: T significantly increased conversion of L-cysteine to H(2)S. This effect was significantly reduced by PGG and BCA, two specific inhibitors of CSE. T (10 nM-10 microM) induced a concentration-dependent vasodilatation of rat aortic rings in vitro that was significantly and concentration-dependent inhibited by PGG, BCA, and glybenclamide. Incubation of aorta with T up to 1 h did not change CBS/CSE expression, suggesting that T modulates enzymatic activity. CONCLUSIONS: Here we demonstrate that T vasodilator effect involves H(2)S, a novel gaseous mediator. T modulates H(2)S levels by increasing the enzymatic conversion of L-cysteine to H(2)S. PMID- 18511181 TI - Editorial comment on: surveillance study in Europe and Brazil on clinical aspects and Antimicrobial Resistance Epidemiology in Females with Cystitis (ARESC): implications for empiric therapy. PMID- 18511182 TI - Editorial comment on: surveillance study in Europe and Brazil on clinical aspects and Antimicrobial Resistance Epidemiology in Females with Cystitis (ARESC): implications for empiric therapy. PMID- 18511183 TI - Good outcome for patients with few lymph node metastases after radical retropubic prostatectomy. AB - BACKGROUND: Conflicting results exist regarding the value of an extended pelvic lymph node dissection (PLND) in node-positive patients undergoing radical retropubic prostatectomy (RRP) for clinically localized prostate cancer. OBJECTIVE: To assess the long-term outcome in node-positive patients who underwent extended PLND followed by RRP. DESIGN, SETTING, AND PARTICIPANTS: A consecutive series of 122 node positive patients with negative preoperative staging examinations, no neoadjuvant hormonal or radiotherapy, and who underwent extended PLND (>/=10 lymph nodes in the surgical specimen) followed by RRP were analyzed. None of the patients received immediate androgen deprivation therapy (ADT). INTERVENTION: All patients underwent extended PLND followed by RRP. MEASUREMENTS: Biochemical recurrence-free survival, cancer-specific, and overall survival were assessed using the Kaplan-Meier technique. RESULTS AND LIMITATIONS: Median prostate-specific antigen (PSA) was 16ng/ml. At pathological examination 76% of the 122 patients had pT3-pT4 tumours, 50% seminal vesicle infiltration. A median of 22 nodes were removed per patient. Median cancer-specific survival at 5 and 10 yr was 84.5% and 60.1%, respectively. In patients with /=3 positive nodes removed, median cancer-specific survival at 10 yr was 78.6% and 33.4%, respectively (p<0.001). After a median period of 33 mo, 61 of the 122 patients (50%) received ADT, particularly those (69%) with >/=3 positive nodes removed. This retrospective study includes a significant percentage of patients with high tumour burden, and therefore may not reflect current patient series. CONCLUSIONS: Patients with /=3 positive nodes, despite extended PLND and despite ADT in 69% of patients. PMID- 18511184 TI - Is node-positive prostate cancer always a systemic disease? PMID- 18511185 TI - Preliminary kinetic study on the degradation of nitrobenzene by modified ceramic honeycomb-catalytic ozonation in aqueous solution. AB - The kinetics of degradation of nitrobenzene in aqueous solution was investigated in the processes of ozone alone, ozone/ceramic honeycomb (CH), ozone/modified ceramic honeycomb (MCH). The results indicated that all reactions followed the pseudo-first-order kinetic model, and the degradation rate of nitrobenzene was accelerated in the presence of CH or MCH catalyst, and the more pronounced degradation rate was achieved in O(3)/MCH system. Under the experimental conditions of reaction temperature 293 K and initial pH 6.87, the rate constants were determined to be 5.21 x 10(-2)min(-1) for O(3) alone, 7.99 x 10(-2)min(-1) for O(3)/CH and 15.45 x 10(-2)min(-1) for O(3)/MCH. The influencing factors, such as applied ozone concentration (0.987-2.732 mg L(-1)), initial concentration of nitrobenzene (50-250 microg L(-1)) and amount of catalyst (0-5 blocks) could yield respectively the positive effect on the pseudo-first-order rate constants for degradation of nitrobenzene in the three processes mentioned above. The results suggested that the modification process promoted the catalytic activity of raw CH catalyst, namely the impregnation of metals (Mn, Cu and K) maybe enhance the initiation of hydroxyl radical (OH). PMID- 18511186 TI - Ozonation of trichloroethylene in acetic acid solution with soluble and solid humic acid. AB - The combined flushing and oxidation process using acetic acid and ozone has been used successfully to remove trichloroethylene (TCE) completely from contaminated soil. In this study, the effects of humic acid, a fraction of the organic matter in soil, over the performance of TCE decomposition was evaluated. TCE decomposition by ozone was enhanced by the presence of humic acid at concentrations lower than 8mgCL(-1) and then inhibited at higher concentrations. It is possible that the presence of the soluble humic acid fraction during the ozonation of TCE in acetic acid solutions produces hydroxyl radicals during the TCE ozonation which appears to be the reason for the enhanced TCE decomposition rate. Solid humic acid reduced TCE decomposition rate by acting as an ozone scavenger. Similarly, sorbed TCE reduced the amount of TCE available for decomposition by ozone in solution. PMID- 18511187 TI - Microbial and nutrient stabilization of two animal manures after the transit through the gut of the earthworm Eisenia fetida (Savigny, 1826). AB - Here we studied how the transit through the gut of the earthworm Eisenia fetida affects the microbial and nutrient stabilization of pig and cow manure, by analyzing fresh casts. Earthworms reduced the pools of dissolved organic C and N in casts from both types of manure, as wells as mineral N. Microbial biomass was enhanced only in casts from pig manure and did not change in casts from cow manure, and fungal populations only raised in casts from cow manure. Earthworms reduced microbial activity in casts from cow manure and did not modify in casts from pig slurry. Enzyme activities in casts also depended on the manure ingested; there were no changes in dehydrogenase and beta-glucosidase activities, whereas acid and alkaline phosphatases increased. The results indicate that the first stage in vermicomposting of pig and cow manure by E. fetida, i.e. casting, produced a microbial stabilization decreasing the activity of microorganisms; this stabilization occurred despite of the increase in microbial biomass. The strong reduction in nutrient pools of manures may be the responsible of this contradiction. These changes will influence the dynamics of the organic matter degradation by reducing forms of C and N available to microorganisms and hence restricting their growth and multiplication. Nevertheless, casts were also characterized by an increased enzyme potential that might lead to a further thorough degradation of pig and cow manure. PMID- 18511188 TI - Novel method to evaluate the net wear volume of bag-filter by fly ash. AB - In order to study the wear of bag-filter by ash dust, sintered magnetite (Fe(3)O(4)) compact was used for counter material for sliding wear test. The precise amount of magnetite particles embedded into bag-filter was determined by a vibration sample magnetometer (VSM) measurement. It is found that net amount of wear of the bag-filter could be precisely determined by the magnetic measurement. It is also found that the net amount of sliding wear of the bag-filter increases with increasing the wear distance, sliding speed and applied load. To discuss the validity of proposed method, shot peening test with deoxidized iron particles was also carried out for bag-filter sample. The data obtained by the shot peening test is consistent with that obtained by the sliding wear test. The proposed method is, thus, very useful to evaluate the net amount of wear of bag-filter by the fly ash. PMID- 18511189 TI - Ammonia removal in electrochemical oxidation: mechanism and pseudo-kinetics. AB - This paper investigated the mechanism and pseudo-kinetics for removal of ammonia by electrochemical oxidation with RuO(2)/Ti anode using batch tests. The results show that the ammonia oxidation rates resulted from direct oxidation at electrode liquid interfaces of the anode by stepwise dehydrogenation, and from indirect oxidation by hydroxyl radicals were so slow that their contribution to ammonia removal was negligible under the condition with Cl(-). The oxidation rates of ammonia ranged from 1.0 to 12.3 mg N L(-1)h(-1) and efficiency reached nearly 100%, primarily due to the indirect oxidation of HOCl, and followed pseudo zero order kinetics in electrochemical oxidation with Cl(-). About 88% ammonia was removed from the solution. The removed one was subsequently found in the form of N(2) in the produced gas. The rate at which Cl(-) lost electrons at the anode was a major factor in the overall ammonia oxidation. Current density and Cl(-) concentration affected the constant of the pseudo zero-order kinetics, expressed by k=0.0024[Cl(-)]xj. The ammonia was reduced to less than 0.5 mg N L(-1) after 2h of electrochemical oxidation for the effluent from aerobic or anaerobic reactors which treated municipal wastewater. This result was in line with the strict discharge requirements. PMID- 18511190 TI - Electrochemical destruction of dinitrotoluene isomers and 2,4,6-trinitrotoluene in spent acid from toluene nitration process. AB - Mineralization of dinitrotoluene (DNT) isomers and 2,4,6-trinitrotoluene (TNT) in spent acid was conducted by in situ electrogenerated hydrogen peroxide. The electrolytic experiments were carried out to elucidate the influence of various operating parameters on the performance of mineralization of total organic compounds (TOC) in spent acid, including electrode potential, reaction temperature, oxygen dosage and concentration of sulfuric acid. It is worth noting that organic compounds could be completely mineralized by hydrogen peroxide obtained from cathodic reduction of oxygen, which was mainly supplied by anodic oxidation of water. Based on the spectra identified by gas chromatograph/mass spectrometer (GC/MS), it is proposed that oxidative degradation of 2,4-DNT and/or 2,6-DNT, 2,4,6-TNT results in o-mononitrotoluene (MNT) and 1,3,5-trinitrobenzene, respectively. Due to the removal of TOC and some amount of water, the electrolytic method established is promising for industrial application to regeneration of spent acid from toluene nitration process. PMID- 18511191 TI - Effects of hindlimb unloading and reloading on c-fos expression of spinal cord evoked by vibration of rat Achille tendon. AB - To determine whether the neuronal activity of the spindle cord in muscle spindle afferent pathways is altered after a period of hindlimb unloading (Hu), and after recovery, we focused on c-Fos-immunoreactivity in the spinal cord evoked by excitation of the muscle spindle in normal gravity and after 3, 7, and 14 days of Hu, and after 14 days of Hu with an additional 2, 5, and 9 days of unrestricted cage activity. High frequency sinusoidal vibration (HFV) applied to the Achilles tendon was used for activation of the muscle spindle. Results showed that c-Fos immunoreactive neurons evoked by HFV were mainly concentrated in lamina IV-VII of the ipsilateral spinal cord. After 14 days of Hu, the total number of labeled neurons in the spinal cord was significantly increased (P<0.05). The number of c Fos-immunoreactive neurons in lamina IV and VI-VII were also significantly higher than that in normal rats (P<0.05). After 9 days of reloading, the total number of c-Fos-immunoreactive neurons and the pattern of their lamellar distribution were both recovered to pre-experimental levels. Our findings suggest that simulated weightlessness induces a large increase in neuronal excitation of the spinal cord in muscle spindle afferent pathways. Weight bearing immediately after Hu could induce completed recovery of excitability of the spinal cord in muscle spindle afferent pathways. PMID- 18511192 TI - Differential regulation of chondroitin sulfate proteoglycan mRNAs in the denervated rat fascia dentata after unilateral entorhinal cortex lesion. AB - Following brain trauma, chondroitin sulphate proteoglycans (CSPGs) are enriched at injury sites and in denervated areas. At injury sites, CSPGs are regarded as inhibitors of axonal regeneration because of their growth inhibitory properties. In areas of denervation their role is less clear, since they are enriched in zones of sprouting, i.e. zones of axonal growth. To identify CSPGs expressed in a denervated brain area and to quantify changes in their mRNA expression, neurocan, brevican, NG2, phosphacan and aggrecan mRNA were analyzed in the rat fascia dentata following entorhinal denervation. Laser microdissection was combined with quantitative RT-PCR to measure mRNA changes specifically within the denervated portion of the molecular layer (1h, 6h, 10h, 12h, 1d, 2d, 3d, 4d, 7d and 14d post lesion). Changes in glial fibrillary protein mRNA were measured at the same time points and used as lesion control. This approach revealed a differential regulation of CSPG mRNAs in the denervated zone: neurocan, brevican and NG2 mRNA were upregulated with a maximum around 2 days post-lesion. In contrast, aggrecan mRNA levels reached a maximum 7 days post-lesion and phosphacan mRNA levels were not significantly altered. Taken together, our data reveal a temporal pattern in CSPG mRNA expression in the denervated fascia dentata. This suggests specific biological functions for CSPGs during the denervation-induced reorganization process: whereas the early increase in CSPGs in the denervated zone could influence the pattern of sprouting, the late increase of aggrecan mRNA suggests a different role during the late phase of reorganization. PMID- 18511193 TI - The effect of viscous loading on brain ependymal cilia. AB - Ependymal cilia line the ventricular system moving cerebral spinal fluid close to the brain surface. They may be exposed to fluid of increasing viscosity in certain pathological conditions such as bacterial meningitis. Our aim was to determine the effect of increasing viscosity on ciliary function. Ciliated ependyma was exposed to solutions of different viscosities (1-60cP) and ciliary function assessed by high-speed digital imaging. The mean (S.D.) ciliary beat frequency (CBF), measured after 30min incubation in Medium 199 at 37 degrees C, was 34.9 (2.9)Hz. Increased viscous loading was followed by a rapid decrease in CBF compared to baseline readings (p<0.001). After 15min of exposure to the increased viscous load, CBF reached a new stable level while the viscous load was maintained. Compared to baseline measurements of CBF, viscous loading of 3.7cP caused a 16%, 10.4cP at 34% and 24cP a 70% decrease in beat frequency. Further viscous loading at levels up to 60cP resulted in no further reduction of ependymal CBF. Solutions of 24 and 40cP had no effect on ciliary amplitude. An increase in viscosity to 60cP caused a significant (30%: p=0.001) decrease in the ciliary beat amplitude. PMID- 18511194 TI - Simulation of rabbit A-type retinal horizontal cell that generates repetitive action potentials. AB - Rabbit A-type retinal horizontal cells are classified into at least two types: cells that generate repetitive action potentials and those that do not. A mathematical model of these two types of cells based on the ionic current mechanisms has been proposed. Although the response of the former cell to 1-s positive current injection has so far been investigated by both electrophysiological and computational approaches, how this cell responds to much longer current injection has not been investigated. In the present study, I use this model to investigate the response of the former cell to a much longer current injection. Computer simulation indicates that when the stimulation period is relatively short, the membrane potential returns to the resting potential after current injection. In this case, the cell can repeatedly generate repetitive action potentials in response to stimulation. In contrast, when the stimulation period is relatively long, the membrane potential does not return to the resting potential but maintains the depolarized level even after current injection. In this case, the cell cannot repeatedly generate repetitive action potentials. When the present results were compared with those of the previous study, the difference in the response pattern after stimulation between the two types of horizontal cells was revealed. PMID- 18511195 TI - Combination of the serotonin transporter and norepinephrine transporter gene promoter polymorphisms might influence harm avoidance and novelty seeking in healthy females. AB - It has been demonstrated that the interaction between serotonin transporter (5HTT) and norepinephrine transporter (NET) functions affects each transporter function and behavior in studies using knockout mice model. In the present study, we examined the effects of the 5HTT and NET gene promoter polymorphisms on personality traits in 575 Japanese healthy subjects. The 5HTT (long/short, L/S) and NET (-3081 A/T) genotypes were identified by PCR methods, and personality traits were assessed by the Temperament and Character Inventory (TCI). Neither of the two polymorphisms affected any TCI dimension, but the interaction between them had significant effects on harm avoidance and novelty seeking in females. Subsequent analyses showed that the females with the combination of the SS genotype reducing 5HTT function and the TT genotype reducing NET function had higher harm avoidance and lower novelty seeking. The present study suggests that the combination of 5HTT and NET polymorphisms influences harm avoidance and novelty seeking in females. PMID- 18511196 TI - Withdrawal-like behavior in planarians is dependent on drug exposure duration. AB - Planarians display a concentration-related reduction in locomotor activity following their spontaneous withdrawal from opioids, cannabinoids, stimulants and benzodiazepines. This suggests that planarians display a withdrawal-like behavior that can be quantified as a reduction in locomotor activity. Because withdrawal like behavior in previous studies has been quantified only following the cessation of a 60-min drug exposure, it is unclear whether the withdrawal response varies with drug exposure duration. Therefore, the goal of this study is to determine if the duration of drug exposure (0, 5, 15, 30, 45, 60min and 24h) to three different drugs - methamphetamine, cocaine and caffeine - affects the magnitude of withdrawal-like behavior (i.e., reduced locomotor activity) in planarians. Experiments revealed that methamphetamine (10microM) produced significant withdrawal-like behavior regardless of the exposure time (P<0.05). An exposure time of only 5min resulted in a significant reduction in locomotor activity. The peak effect, although occurring following a 24-h exposure, was only slightly greater than that caused by a 30-min exposure. For cocaine (10microM), a longer exposure time (15min) was required for the manifestation of significant withdrawal-like behavior. The peak cocaine effect was observed following a 24-h exposure. Caffeine (10microM) did not produce significant changes in locomotor activity during withdrawal or alter locomotor activity during acute exposure. The present results suggest that the magnitude of withdrawal-like behavior in planarians is dependent on both the duration and type of drug exposure, and that planarians do not display withdrawal to caffeine. PMID- 18511197 TI - Activation of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) during hypoxia in cerebral cortical nuclei of guinea pig fetus at term: role of nitric oxide. AB - Previously we have shown that cerebral tissue hypoxia results in generation of nitric oxide (NO) free radicals as well as increased expression of mitogen activated protein kinase like extracellular signal-regulated kinase (ERK) and c jun N-terminal kinase (JNK). The present study tested the hypothesis that administration of l-nitro-l-arginine methyl ester (L-NAME), a NOS inhibitor, prior to hypoxia prevents the hypoxia-induced activation of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) and in the cerebral cortex of the term guinea pig fetus. To test this hypothesis normoxic (Nx, n=6), hypoxic (Hx, n=7) and hypoxic pretreated with l-NAME (Hx+L-NAME, n=6) guinea pig fetuses at 60 days gestation were studied to determine the phosphorylated p38, ERK and JNK. Hypoxia was induced by exposing pregnant guinea pigs to FiO2 of 0.07 for 1h. l-NAME (30mg/kg i.p.) was administered to pregnant mothers 60min prior to hypoxia. Cerebral tissue hypoxia was documented biochemically by determining the tissue levels of ATP and phosphocreatine (PCr). Neuronal nuclei were isolated, purified and proteins separated using 12% SDS-PAGE, and then probed with specific phosphorylated ERK, JNK and p38 antibodies. Protein bands were detected by enhanced chemiluminescence, analyzed by imaging densitometry and expressed as absorbance (ODxmm2). The relative level of p-p38 was 51.41+/-9.80 (Nx), 173.67+/ 3.63 (Hx), 58.56+/-3.40 (Hx+L-NAME), p<0.05 vs. Hx. The relative level p-ERK was 44.91+/-4.20 (Nx), 135.12+/-17.02 (Hx), 58.37+/-9.5 (Hx+L-NAME), p<0.05 vs. Hx. The relative level of p-JNK was 34.86+/-6.77 (Nx), 97.36+/-19.24 (Hx), 46.65+/ 12.81 (Hx+L-NAME), p<0.05 vs. Hx. The data show that administration of l-NAME prior to hypoxia decreased the relative level of phosphorylated p38, ERK and JNK at term gestation. Since a NOS inhibitor prevented the hypoxia-induced phosphorylation of p38, ERK and JNK, we conclude that the hypoxia-induced activation of p38, ERK and JNK in the cerebral cortical nuclei of guinea pig fetus at term is NO-mediated. We speculate that NO-mediated modification of cysteine residue leading to inhibition of MAP kinase phosphatases results in increased activation of p38, ERK and JNK in the guinea pig fetus at term. PMID- 18511198 TI - Projections from the vestibular nuclei and nucleus prepositus hypoglossi to dorsal raphe nucleus in rats. AB - The serotonergic system regulates processing in components of the vestibular nuclear complex, including the medial vestibular nucleus (MVe) and nucleus prepositus hypoglossi (PH). Recent studies using anterograde and retrograde tracers have shown that vestibular nuclei are targeted by regionally selective projections from the serotonergic dorsal raphe nucleus. The objective of the present investigation was to determine whether the DRN is targeted by projections from the vestibular nuclear complex in rats, using the anterograde tracer biotinylated dextran amine (BDA). After injection of BDA into PH or the caudal parvicellular division of MVe, labeled fibers and terminals were observed in the ventromedial and lateral subdivisions of DRN. These findings indicate that projections from the vestibular nuclei and PH are organized to modulate processing within specific functional domains of the DRN. PMID- 18511199 TI - Tegumental surface changes in juvenile Fasciola hepatica in response to treatment in vivo with triclabendazole. AB - Eight indoor-reared crossbred sheep with no pre-exposure to Fasciola hepatica were infected, by oral gavage, with 200 metacercarial cysts of the triclabendazole (TCBZ)-susceptible Cullompton isolate of F. hepatica. Anthelmintic dosing occurred at 4 weeks post-infection with 10 mg/kg triclabendazole. Two treated sheep were euthanized at 48 h, 72 h and 96 h post treatment with triclabendazole. Two control sheep were euthanized alongside the 48 h triclabendazole-treated sheep. Juvenile flukes were recovered from each of the sheeps' liver and processed for scanning electron microscopy (SEM). Flukes were still active 48 h post-treatment and displayed limited morphological disruption. There was some blebbing and sloughing of the tegument around the oral sucker. In several of the specimens, an extra layer had been deposited on the fluke surface, giving it a flattened appearance. At 72 h post-treatment, only one fluke remained alive and the disruption varied in degree. In the majority of flukes, there was severe swelling of the tegument, accompanied by isolated areas of flattening along the lateral margins of the flukes and in the tail region. Limited areas of sloughing occurred in the tail region. In more seriously affected specimens, the syncytium had been stripped away to reveal the basal lamina and some deeper lesions were also observed. By 96 h post-treatment, all the flukes were dead and were grossly disrupted. They were totally devoid of tegument and deep lesions exposed the internal tissues of the fluke. PMID- 18511200 TI - PTSD in persons with burns: an explorative study examining relationships with attributed responsibility, negative and positive emotional states. AB - A sample of 90 persons who had been hospitalized for severe burns were interviewed 1-4 years after the incident. Current DSM-IV post-traumatic stress disorder (PTSD) was assessed with the Composite International Diagnostic Interview. Perceived attributed responsibility and related positive and negative emotional states were examined using a semi-structured interview. Findings showed that PTSD was established in 8% of the participants and partial PTSD in 13%. In a homogeneity analysis (HOMALS), PTSD was associated with the attribution of responsibility for the incident to impersonal relationships and with a negative emotional state. The absence of (partial) PTSD was associated with the attribution of responsibility to close relationships, internal and circumstance related attribution of responsibility and neutral or forgiving feelings. In logit analyses, both emotional state as well as attributed responsibility are significantly related to (partial) PTSD. However, the model including emotional state showed to have the best fit. Although further research is needed, these results may indicate that professionals working in burn care should consider the emotional state in relation to perceived attribution of responsibility when considering PTSD. Promoting forgiveness may be a beneficial strategy in dealing with post-traumatic stress reactions. PMID- 18511201 TI - Psychological impact of burns on children treated in a severe burns unit. AB - The aims of this study were to analyse the types of incident which cause children's burns, to investigate emotional reactions and associated behaviour afterward and to ascertain post-admission sequelae. A total of 83 young people took part, aged 1-17 years, with a mean 12% (range 1.5-47%, S.D. 8.0) total body surface area burned. According to the results from the psychometric scales and questionnaire used, most burns were the result of domestic incidents. Older children scored higher in anxiety. The most common behavioural reactions observed (attachment, wish to go back home) reflect the particularity of these patients compared with other samples of children in hospital. With respect to post admission sequelae, no relevant results were obtained. PMID- 18511202 TI - Assessment of burn depth and burn wound healing potential. AB - The depth of a burn wound and/or its healing potential are the most important determinants of the therapeutic management and of the residual morbidity or scarring. Traditionally, burn surgeons divide burns into superficial which heal by rapid re-epithelialization with minimal scarring and deep burns requiring surgical therapy. Clinical assessment remains the most frequent technique to measure the depth of a burn wound although this has been shown to be accurate in only 60-75% of the cases, even when carried out by an experienced burn surgeon. In this article we review all current modalities useful to provide an objective assessment of the burn wound depth, from simple clinical evaluation to biopsy and histology and to various perfusion measurement techniques such as thermography, vital dyes, video angiography, video microscopy, and laser Doppler techniques. The different needs according to the different diagnostic situations are considered. It is concluded that for the initial emergency assessment, the use of telemetry and simple burn photographs are the best option, that for research purposes a wide range of different techniques can be used but that, most importantly, for the actual treatment decisions, laser Doppler imaging is the only technique that has been shown to accurately predict wound outcome with a large weight of evidence. Moreover this technique has been approved for burn depth assessment by regulatory bodies including the FDA. PMID- 18511203 TI - Developmental exposure to 3,4-methylenedioxymethamphetamine results in downregulation of neurogenesis in the adult mouse hippocampus. AB - 3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a powerful releaser of 5-HT and chronic use of this drug can cause depletion of monoamines. Recently, concerns about the risk of adult brain damage due to fetal exposure to MDMA have been raised. We investigated whether developmental MDMA exposure affected adult neurogenesis in C57 black/6 mice. MDMA (1.25 or 20 mg/kg, p.o.) or vehicle was administered daily to the mother from prenatal 6th day to postnatal 21st day. When the offspring were 11 weeks old, they were injected with 5-bromo-2' deoxyuridine (BrdU) (120 mg/kg, i.p.) once a day for 4 days. After 24 h or 28 days, the animals were killed to count the BrdU-positive cells in the dentate gyrus. At 24 h after the last BrdU injection, the number of BrdU-positive cells in the offspring developmentally exposed to MDMA was significantly lower than that of the control group. At 28 days post-BrdU labeling, BrdU-positive cells in the dentate gyrus of female offspring with developmental exposure to high dose MDMA were significantly fewer compared with the control group. In addition, most BrdU-positive cells were co-labeled with the mature neuronal marker, neuronal nuclei, while a few BrdU-labeled cells were merged with an astrocyte marker. Our results suggest that developmental exposure to MDMA can result in decreases in the proliferation and survival of mature newborn cells in the adult dentate gyrus. PMID- 18511204 TI - Neonatal 6-hydroxydopamine lesions of the frontal cortex in rats: persisting effects on locomotor activity, learning and nicotine self-administration. AB - Dopaminergic innervation of the frontal cortex in adults is important for a variety of cognitive functions and behavioral control. However, the role of frontal cortical dopaminergic innervation for neurobehavioral development has received little attention. In the current study, rats were given dopaminergic lesions in the frontal cortex with local micro-infusions of 6-hydroxydopamine (6 OHDA) at 1 week of age. The long-term behavioral effects of neonatal frontal cortical 6-OHDA lesions were assessed in a series of tests of locomotor activity, spatial learning and memory, and i.v. nicotine self-administration. In addition, neurochemical indices were assessed with tissue homogenization and HPLC in the frontal cortex, striatum, and nucleus accumbens of neonatal and adult rats after neonatal 6-OHDA lesions. In neonatal rats, frontal 6-OHDA lesions as intended caused a significant reduction in frontal cortical dopamine without effects on frontal cortical 5-HT and norepinephrine. The frontal cortical dopamine depletion increased 5-HT and norepinephrine levels in the nucleus accumbens. Locomotor activity assessment during adulthood in the figure-8 maze showed that lesioned male rats were hyperactive relative to sham-lesioned males. Locomotor activity of female rats was not significantly affected by the neonatal frontal 6-OHDA lesion. Learning and memory in the radial-arm maze was also affected by neonatal frontal 6-OHDA lesions. There was a general trend toward impaired performance in early maze acquisition and a paradoxical improvement at the end of cognitive testing. Nicotine self-administration showed significant lesion x sex interactions. The sex difference in nicotine self-administration with females self-administering significantly more nicotine than males was reversed by neonatal 6-OHDA frontal cortical lesions. Neurochemical studies in adult rats showed that frontal cortical dopamine and DOPAC levels significantly correlated with nicotine self administration in the 6-OHDA-lesioned animals but not in the controls. Frontal cortical 5-HT and 5HIAA showed inverse correlations with nicotine self administration in the 6-OHDA-lesioned animals but not in the controls. These results show that interfering with normal dopamine innervation of the frontal cortex during early postnatal development has persisting behavioral effects, which are sex-specific. PMID- 18511205 TI - Apocynin improves outcome in experimental stroke with a narrow dose range. AB - Inflammation following ischemic stroke is known to contribute to injury. NADPH oxidase (NOX) is a major enzyme system originally studied in immune cells that leads to superoxide (O.*) generation. Apocynin is a NOX inhibitor that has been studied as a potential treatment in experimental stroke. Here we explored the effect of different doses of apocynin in a mouse model of 2 h transient middle cerebral artery occlusion (tMCAO) followed by 22 h reperfusion. Apocynin, given i.v. at a dose of 2.5 mg/kg 30 min before reperfusion, improved neurological function (P<0.01), reduced infarct volume (P<0.05), and reduced the incidence of cerebral hemorrhage (P<0.05), but not at higher doses of 3.75 and 5 mg/kg, where it actually increased brain hemorrhage. Apocynin also tended to reduce mortality at the lower dose, but not at higher doses. Using hydroethine fluorescence to delineate O.* in the brain, neurons and some microglia/macrophages, but not vascular endothelial cells were found to contain O.*. Apocynin at protective doses markedly prevented ischemia-induced increases in O.*. Our data suggested that apocynin can protect against experimental stroke, but with a narrow therapeutic window. PMID- 18511206 TI - Hippocampal levels of gamma-enolase, C-1-tetrahydrofolate synthase and serotransferrin fluctuate over the estrous cycle in the rat. AB - Hippocampal functions vary across the estrous cycle but metabolic changes at the protein level have not been systematically studied so far. It was therefore the aim of the study to screen expression of metabolic proteins mainly represented by metabolic enzymes in the hippocampus over the estrous cycle and in males. Female and male OFA Sprague-Dawley rats were used and female estrous phases were determined by vaginal smears, according to which females were separated into groups of proestrous, estrous, early and late metestrous and diestrous. Proteins were extracted from hippocampal tissue and separated on two-dimensional gel electrophoresis followed by identification with mass spectrometry methods (MALDI TOF-TOF and nano-LC-ESI-MS/MS). Comparative analysis of protein levels was carried out by quantifying protein spot volumes by means of specific software. Levels of one expression form of gamma-enolase were different between diestrous and early metestrous; C-1-tetrahydrofolate synthase levels were elevated in proestrous as compared with estrous and serotransferrin levels were increased in diestrous as compared with proestrous, estrous, metestrous and in males. The outcome of estrous cycle- and gender-dependent protein fluctuations is relevant for the interpretation of previous and future work as well as for the design of further studies at the protein level in the hippocampus. PMID- 18511207 TI - Inhibition of spinal cytosolic phospholipase A(2) expression by an antisense oligonucleotide attenuates tissue injury-induced hyperalgesia. AB - Activation of the spinal phospholipase A(2) (PLA(2)) -cyclooxygenase (COX) prostaglandin signaling pathway is widely implicated in nociceptive processing. Although the role of spinal COX isoforms in pain signal transmission has been extensively characterized, our knowledge of PLA(2) enzymes in this cascade is limited. Among all PLA(2) groups, cytosolic calcium-dependent PLA(2) group IVA (cPLA(2)IVA) appears to be the predominant PLA(2) enzyme in the spinal cord. In the present study we sought to (i) characterize anatomical and cellular distribution and localization of cPLA(2)IVA in dorsal horn of rat spinal cord, (ii) verify efficacy and selectivity of intrathecal (IT) delivery of an antisense oligonucleotide (AS) targeting rat cPLA(2)IVA mRNA on spinal expression of this enzyme, and (iii) examine the effect of down-regulation of spinal cPLA(2)IVA on peripheral tissue injury-induced pain behavior. Here we demonstrate that cPLA(2)IVA is constitutively expressed in rat spinal cord, predominantly in dorsal horn neurons and oligodendrocytes but not in astrocytes or microglia. Intrathecal injection of AS significantly down-regulated both protein and gene expression of cPLA(2)IVA in rat spinal cord, while control missense oligonucleotide (MS) had no effect. Immunocytochemistry confirmed that the reduction occurred in neurons and oligodendrocytes. cPLA(2)IVA AS did not alter expression of several other PLA(2) isoforms, such as secretory PLA(2) (groups IIA and V) and calcium-independent PLA(2) (group VI), indicating that the AS was specific for cPLA(2)IVA. This selective knockdown of spinal cPLA(2)IVA did not change acute nociception (i.e. paw withdrawal thresholds to acute thermal stimuli and intradermal formalin-induced first phase flinching), however, it significantly attenuated formalin-induced hyperalgesia (i.e. second phase flinching behavior), which reflects spinal sensitization. Thus the present findings suggest that cPLA(2)IVA may specifically participate in spinal nociceptive processing. PMID- 18511208 TI - Double-stranded RNA-mediated suppression of Period2 expression in the suprachiasmatic nucleus disrupts circadian locomotor activity in rats. AB - Circadian behavioral rhythms in mammals are controlled by a central clock located in the suprachiasmatic nucleus (SCN). PER2, the protein product of the clock gene, Period 2 (Per2), is expressed rhythmically in the SCN [Beaule C, Houle LM, Amir S (2003) Expression profiles of PER2 immunoreactivity within the shell and core regions of the rat suprachiasmatic nucleus: Lack of effect of photic entrainment and disruption by constant light. J Mol Neurosci 21:133-148] and has been implicated in the control of circadian behavioral rhythms based on the evidence that genetic mutations in Per2 abolish free running locomotor activity rhythms in mice [Zheng B, Larkin DW, Albrecht U, Sun ZS, Sage M, Eichele G, Lee CC, Bradley A (1999) The mPer2 gene encodes a functional component of the mammalian circadian clock. Nature 400:169-173; Bae K, Jin X, Maywood ES, Hastings MH, Reppert SM, Weaver DR (2001) Differential functions of mPer1, mPer2, and mPer3 in the SCN circadian clock. Neuron 30:525-536]. Such mutations eradicate PER2 expression in the SCN and disrupt the SCN molecular clockwork, however, they also affect PER2 in the rest of the brain and body leaving open the possibility that the changes in behavioral rhythms might be influenced, at least in part, by disruptions in PER2 functioning outside the SCN. We used RNA interference mediated transient knockdown of Per2 to study the effect of selective suppression of PER2 expression in the SCN, per se, on behavioral circadian rhythms. We found that transient suppression of PER2 in the SCN disrupted free running locomotor activity rhythms for up to 10 days in rats. Infusions of control dsRNA into the SCN or infusions of dsRNA to Per2 immediately dorsal to the SCN had no effect. These results constitute evidence for a direct link between PER2 expression in the SCN and the expression of behavioral circadian rhythms in mammals. PMID- 18511210 TI - Status epilepticus induces a TrkB to p75 neurotrophin receptor switch and increases brain-derived neurotrophic factor interaction with p75 neurotrophin receptor: an initial event in neuronal injury induction. AB - In neuronal cultures it has been demonstrated that neurotrophins can elicit neuronal death through the p75 neurotrophic receptor (p75ntr) in the absence of concomitant Trk signaling. However, it was suggested that p75ntr induces neuronal death after status epilepticus (SE) in neuronal populations that express relatively high quantities of tropomyosin receptor kinase B (TrkB). Here, using Western blot and immunohistochemistry analyses in the hippocampus, we found that 3-h SE caused a remarkable decrease in TrkB expression and phosphorylation, and a significant increase in p75ntr. TrkB modification occurs before the overexpression of the tumor suppressor protein p53, accompanies the cell damage taking place in the dentate gyrus, and precedes the CA1 neuronal injury as assessed by Fluoro-Jade B and Nissl staining. Co-immunoprecipitation of brain derived neurotrophic factor (BDNF) or its immature form proBDNF showed increased interaction with p75ntr after its binding to TrkB was reduced. Interestingly, proBDNF also increases its binding with p75ntr after seizures that do not cause neuronal death (animals injected with pilocarpine that fail to enter SE). However, in those animals, TrkB protein levels remained unchanged and its phosphorylation increased. Our results indicate an intrinsic capacity of neurons in vivo to modify final neurotrophin output by changing the proportion of their receptors' expression and the receptors' interaction with their ligands. These early events support the idea that neurotrophins may be involved in the induction of neuronal death in vivo under pathological conditions. PMID- 18511209 TI - The shell of the nucleus accumbens has a higher dopamine response compared with the core after non-contingent intravenous ethanol administration. AB - Dopamine increases in the nucleus accumbens after ethanol administration in rats, but the contributions of the core and shell subregions to this response are unclear. The goal of this study was to determine the effect of various doses of i.v. ethanol infusions on dopamine in these two subregions of the nucleus accumbens. Male Long-Evans rats were infused with either acute i.v. ethanol (0.5, 1.0, 1.5 g/kg), repeated i.v. ethanol (four 1.0 g/kg infusions resulting in a cumulative dose of 4.0 g/kg), or saline as a control for each condition. Dopamine and ethanol were measured in dialysate samples from each experiment. The in vivo extraction fraction for ethanol of probes was determined using i.v. 4 methylpyrazole, and was used to estimate peak brain ethanol concentrations after the infusions. The peak brain ethanol concentrations after the 0.5, 1.0 and 1.5 g/kg ethanol infusions were estimated to be 20, 49 and 57 mM, respectively. A significant dopamine increase was observed for the 0.5 g/kg ethanol group when collapsed across subregions. However, both the 1.0 g/kg and 1.5 g/kg ethanol infusions produced significant increases in dopamine levels in the shell that were significantly higher than those in the core. An ethanol dose-response effect on dopamine in the shell was observed when saline controls, 0.5, 1.0, and 1.5 g/kg groups were compared. For the cumulative-dosing study, the first, second, and fourth infusions resulted in significant increases in dopamine in the shell. However, these responses were not significantly different from one another. The results of this study show that the shell has a stronger response than the core to i.v. ethanol, that dopamine in the shell increases in a dose-dependent manner between 0.5-1.0 g/kg doses, but that the response to higher ethanol doses reaches a plateau. PMID- 18511211 TI - Recovering at the cost of amnesia? AB - Despite the great progress of psychiatry, many of its fundamental problems remain unresolved. Obviously, new ideas that expand the present understanding of such problems are useful. In this paper, the following hypothesis is proposed: mental disorders are caused by hypoxic phenomena, stimulating hypertrophied growth of a neural network; treatment should be directed towards the damaging of abnormal elements of this network; successful treatment is inevitably accompanied by moderate retrograde amnesia. Based on this hypothesis, the connection between etiology, pathogenesis, and therapy of mental disorders is described, the role of retrograde amnesia is shown, and potential new methods of treatment (shaking, vibration, and ultrasound) are predicted. To test the hypothesis, some means are discussed. The first means is based on the analysis of the reasons of unsuccessful clinical trials of the method of mental disorders treatment by acute hypoxic hypoxia (USA, 1938-1940). Such a method, according to the hypothesis, should damage the abnormal elements of a neural network as a result of acute hypoxia. Analysis of the equipment and procedure shows that the trials were unsuccessful as a result of insufficiently powerful hypoxic influence. Improvements to this method are proposed. When using this method, it is advisable to measure the cerebral blood oxygenation index and reduce it by regulating the oxygen concentration in the hypoxic gas mixture given to the patient. This reduction should continue until the patient reaches a state of moderate retrograde amnesia. It is also advisable to note the blood oxygenation index value at the moment when the patient lapses into unconsciousness, and then, on the basis of this value, estimate a necessary power (i.e., acuteness and duration) of hypoxic hypoxia. Other means for testing the hypothesis are also discussed: retrospective analysis of the results of electroconvulsive therapy, and testing the predicted methods of treatment. PMID- 18511212 TI - The role of altered cerebrospinal fluid dynamics in pathogenesis of multilevel myelopathy with single spinal stenosis. PMID- 18511213 TI - The Wernicke-Kleist-Leonhard "short-circuiting": a missing link between attention deficit hyperactivity disorder, Tourette syndrome, and obsessive-compulsive disorder? AB - According to the Wernicke-Kleist-Leonhard (WKL) School of Psychiatry, human actions could be categorized into spontaneous, expressive, reflex, and reactive. The so-called short-circuit movements represent a pathological exaggeration of immediate reactions to sensory impressions, such as sudden or threatening stimuli. "Short-circuiting" was deemed of utmost importance for the diagnoses of several WKL endogenous psychoses, such as hyperkinetic motility psychosis, periodic catatonia, and proskinetic catatonia. We suggest that short-circuit movements may be an under-reported behavioral phenotype that may cut across different phenotypes of non-psychotic DSM-IV-TR disorders, such as attention deficit hyperactivity disorder (ADHD), Tourette's syndrome (TS), and obsessive compulsive disorder (OCD). Several features of a neuropsychiatric syndrome seen in the coarse frontal lobe disease ("complex psychomotor release phenomena") may provide a neurological working model for "short-circuiting" in ADHD, TS, and OCD. Further, short-circuit movements may be associated with different degrees of serotonergic, dopaminergic, and glutamatergic dysfunctions. Indeed, although there are a growing number of studies reporting successful treatment of short circuit movements with topiramate and other glutamatergic antagonists, treatment response to drugs based on serotonergic or dopaminergic mechanisms of action is more uncertain and, sometimes, detrimental, leading to an aggravation of pathological behavior. A resurgence of the interest on the descriptive psychopathology of the WKL short-circuit movements could provide a new conceptual framework for the studies on the neurobiology of volitional disorders. PMID- 18511214 TI - Leptin: is it a possible specific liver drug for non-alcoholic fatty liver disease (NAFLD)? PMID- 18511215 TI - [Prevent postnatal urinary incontinence by prenatal pelvic floor exercise? Rationale and protocol of the multicenter randomized study PreNatal Pelvic floor Prevention (3PN)]. AB - INTRODUCTION: Female urinary incontinence (UI) is a frequent affection that generates handicap and expenses. There is a link between UI and pregnancy; onset of UI during pregnancy is a risk factor for permanent UI. Postnatal pelvic floor exercise has shown efficacy to improve postnatal UI. However, it remains uncertain if benefits last more than few months. Publication of our rationale for prenatal pelvic floor exercise is an opportunity to expose our pre-specified hypotheses and help health professionals' awareness. OBJECTIVES: The purpose of PreNatal Pelvic floor Prevention (3PN) is to compare the effects of prenatal pelvic floor exercise versus sole written instructions on UI one year after delivery. METHODS AND POPULATION: It is a multicenter, randomized, single blind study. Main inclusion criteria are first, single and non-complicated pregnancy over 18 years. Women randomized in pelvic floor exercise group will undergo eight sessions with a physiotherapist between six and eight months of pregnancy. Our principal criterion is UI score (International Consultation on Incontinence Questionnaire Short Form [ICIQ-SF]) one year after delivery. We plan to include 280 pregnant women in five centers over a 12-month screening period to show a one point difference on UI score. ETHIC AND FINANCING: The study was approved by the IRB Comite de protection des personnes Sud-Ouest et Outre-Mer. It was registered by French Health Products Safety Agency (AFSSAPS) and Clinical Trials.gov. It is supported by the French Ministry of Health through the 2007 Hospital Plan for Clinical Research (PHRC). PERSPECTIVES: We plan to assess if prenatal pelvic floor exercise reduces postnatal medical consultations or physiotherapy sessions. PMID- 18511216 TI - [HIV genetic diversity and its consequences]. AB - Human immunodeficiency viruses HIV-1 and HIV-2 are the results of multi interspecies transmissions from simian virus to humans. HIV-1 viruses are very divergent and are classified in three groups: M, N and O. The group M is subdivided in nine subtypes and numerous Circulating Recombinant Forms. In 1996, protease inhibitors and HAART disposal have modified the prognostic of the HIV infection. However, one of the major problems is the emergence of antiretroviral resistance. A major advance from the last year is the access to antiretroviral in resources limited countries. On the other hand, the development of a vaccine is today hypothetic. PMID- 18511217 TI - Henipaviruses: a new family of emerging Paramyxoviruses. AB - Paramyxoviruses have been implicated in both animal and human infections. Some viruses, such as Morbilliviruses are responsible for large-scale epidemics. However, there are limited observations of these viruses crossing the host species barrier in nature. In 1994, in Australia a fatal infection in horses and humans was identified to be caused by a new Paramyxovirus, Hendra virus (HeV), and in 1998 in Malaysia, a closely related virus, Nipah virus (NiV) was responsible for fatal infections in pigs and humans. These two viruses were sufficiently different from previously described Paramyxoviruses to create a new genus, Henipaviruses. The natural reservoir of these viruses was the fruit bat (Pteropus), which is found in regions extending from the western Pacific to the eastern coast of Africa. Serological studies have established that as many as half the fruit bats in colonies throughout these regions may have antibodies against this family of viruses. The availability of diagnostic reagents for Nipah virus in humans have identified infections in several countries including, Bangladesh, India and Indonesia. In some of these epidemics, mortality in humans exceeds 75%. Deforestation is probably responsible for fruit bats leaving their ecological niches and approaching farms and villages. The infection of humans and animals may occur via contaminated foods or in certain cases by animals to man. At present, only within close families has human-to-human transmission been proposed. Henipavirus infections are probably more widespread than it is at presently known and so it is important to have an intense monitoring for these diseases, especially in countries where large-scale deforestation is happening. PMID- 18511218 TI - Effects of mixed housing of birds from two genetic lines of laying hens on open field and manual restraint responses. AB - Birds from Rhode Island Red (RIR) origin show a lower fear response and less feather pecking than birds from White Leghorn (WL) origin. This study investigated whether responses in fear eliciting tests were affected if RIR and WL birds were housed together. Experimental groups contained either birds from one line only ('pure' groups) or an equal number of RIR and WL birds ('mixed' groups). These arrangements were maintained from hatch onwards, throughout the rearing and laying period. Birds were subjected to open field tests at 5-6 weeks and 17-18 weeks of age and to manual restraint tests at 7-8 weeks and 24 weeks of age. RIR birds were more active in both open field tests and in the manual restraint test at 24 weeks of age as compared with WL birds. RIR birds from pure groups were more active in the open field test at 17-18 weeks and in the manual restraint test at 24 weeks of age than RIR birds from mixed groups. These results suggest that otherwise low fearful RIR birds may adopt a higher fear response if they are housed together with more fearful conspecifics. These effects do not emerge until after 8 weeks of age. PMID- 18511219 TI - Facial hair whorls (trichoglyphs) and the incidence of motor laterality in the horse. AB - Several species demonstrate obvious motor laterality (sidedness, handedness) in their motor function. Motor laterality in the horse affects locomotion and subsequently equine performance during training and may have inherent safety implications for equitation. Some of the most commonly used identification features in the horse are hair whorls (trichoglyphs), since their specific location and character vary to some degree in every horse. We investigated the relationship between the hair flow of single facial hair whorls and the incidence of lateralised motor bias in 219 horses when under saddle in ridden work. The horses exhibited significant differences in motor preferences with 104 left lateralised (LL) horses, 95 right-lateralised (RL) horses compared to only 20 well-balanced (WB) horses (chi(2)=36.9, d.f.=2, P<0.01). There was also a significant difference in the frequency distribution of single facial hair whorl patterns in the horses consisting of 114 horses with counter-clockwise (CC) whorls, 82 horses with clockwise (C) whorls and 23 horses, which had radial (R) whorls (chi(2)=38.87, d.f.=2, P<0.01). Overall there was a statistically significant association between motor behaviour and facial hair whorl patterns in the horses (chi(2)=69.4, d.f.=4, P>0.001). The RL horses had significantly more C facial hair whorls and the LL horses had significantly more CC facial hair whorls than would be expected purely by chance alone (P<0.05). The findings may provide trainers with a useful tool when attempting to identify simple, non-invasive and reliable predictors of motor laterality in the horse. Furthermore, given that efficient targeted training of performance horses during ridden work may produce WB equine athletes, the findings could assist trainers when designing individual specific training programmes for young horses. PMID- 18511220 TI - Identification of novel chicken microRNAs and analysis of their genomic organization. AB - MicroRNAs (miRNAs) represent a family of small noncoding RNAs with important regulatory roles in diverse biological processes ranging from cell differentiation to organism development. In chickens, the full set of miRNAs and the expression patterns of miRNAs during development are still poorly understood when compared to the other vertebrates. In this study, we identified 29 novel miRNAs and 140 potential miRNA loci in the chicken genome by combining the experimental and computational analyses. Detailed expression patterns of 49 miRNAs were first characterized by Northern blotting and indicated the cooperativity of the miRNA expression with their function in embryogenesis and organogenesis. Twenty-seven miRNA clusters were systematically evaluated in the chicken genome and diverse expression patterns for closely linked miRNAs were observed. Our results significantly expand the set of known miRNAs in the chicken and provide the basis for understanding the structural and functional evolution of miRNA genes in vertebrates. PMID- 18511221 TI - A quantitative real-time PCR assay for Ehrlichia ruminantium using pCS20. AB - Heartwater is a tick borne disease that affects ruminants and wild animals in Africa south of the Sahara. It is caused by Ehrlichia ruminantium and transmitted by the tick Amblyomma hebraeum. The protocols currently used to detect heartwater take several days to complete. Here, we describe the development of a pCS20 quantitative real-time PCR TaqMan probe assay to detect E. ruminantium in livestock blood and ticks from the field. The assay is based on the conserved pCS20 gene region of E. ruminantium that contains two overlapping genes, rnc and ctaG [Collins, N.E., Liebenberg, J., De Villiers, E.P., Brayton, K.A., Louw, E., Pretorius, A., Faber, F.E., Van Heerden, H., Josemans, A., Van Kleef, M., Steyn, H.C., Van Strijp, M.F., Zweygarth, E., Jongejan, F., Maillard, J.C., Berthier, D., Botha, M., Joubert, F., Corton, C.H., Thomson, N.R., Allsopp, M.T., Allsopp, B.A., 2005. The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. PNAS 102, 838-843]. The pCS20 quantitative real-time PCR TaqMan probe was compared to the currently used pCS20 PCR and PCR/32P-probe test with regards to sensitivity, specificity and the ability to detect DNA in field samples and in blood from experimentally infected sheep. This investigation showed that the pCS20 quantitative real-time PCR TaqMan probe was the most sensitive assay detecting seven copies of DNA/mul of cell culture. All three assays, however, cross react with Ehrlichia canis and Ehrlichia chaffeensis. The pCS20 real-time PCR detected significantly more positive field samples. Both the PCR and pCS20 real-time PCR could only detect E. ruminantium parasites in the blood of experimentally infected sheep during the febrile reaction. The PCR/32P-probe assay, however, detected the parasite DNA 1 day before and during the febrile reaction. Thus, because this new quantitative pCS20 real-time PCR TaqMan probe assay was the most sensitive and can be performed within 2h it is an effective assay for epidemiological surveillance and monitoring of infected animals. PMID- 18511222 TI - Prototheca zopfii genotypes isolated from cow barns and bovine mastitis in Japan. AB - This study is the first investigation on Japanese isolates of Prototheca zopfii from bovine mastitis and the cow-barn surroundings by molecular characterization to clarify routes of infection for bovine protothecal mastitis. We performed isolation of Prototheca from cow-barn surroundings (drinking water, sewage and feces) and milk samples from cases of bovine mastitis. Genotypes of the 32 isolates of P. zopfii from cow-barn surroundings and 67 isolates from mastitis were analyzed by genotype-specific PCR assays and restriction fragment length polymorphism (RFLP) assays. All mastitis isolates were identified as P. zopfii genotype 2. Conversely, 29 isolates from cow-barn surroundings were identified as P. zopfii genotypes 1 and 3 isolates as genotype 2, respectively. Given these results, both genotypes of P. zopfii could exist in cow-barn surroundings, but no sites were identified as frequent sources of P. zopfii genotype 2. P. zopfii isolates should thus be further explored with regard to genotype to clarify the reservoir of etiological agents in bovine Prototheca mastitis. PMID- 18511223 TI - Changes in unsettled infant sleep and maternal mood following admission to a parentcraft residential unit. AB - BACKGROUND: Maternal mood disorders are common in the first postnatal year, particularly if the infant has sleep problems. AIMS: This study explored the effectiveness of a residential parentcraft unit's treatment for infant sleep problems, and also its impact on the mother's depression, anxiety, and bonding. STUDY DESIGN: A one-group pretest-posttest design with follow-up was used. Assessments were conducted while participants were at the service, and at 5 weeks and 4 months after discharge. PARTICIPANTS: Mothers (n=116) with infants 3 weeks to 3 years old presenting to a five-day program at an Australian residential parentcraft service because of unsettled infant sleep were recruited. OUTCOME MEASURES: The Edinburgh Postnatal Depression Scale, the Hospital Anxiety and Depression Scale-Anxiety subscale (maternal mood), the Being a Mother & Bonding Scale (maternal experience and mother-infant bonding), and a Change Questionnaire (for infant sleep and maternal emotional health). RESULTS: By 5 weeks post discharge significant improvements were reported for infant sleep behavior, maternal mood (depressive and anxious symptoms), and maternal-infant bonding. 80% of the women reported an improvement on both their infants' sleep and their own emotional health, with strategies and support provided in the unit being cited by most women as the reason for improvement. Emotional health improvements were most often attributed to the improvement in the infant's sleep. All improvements by 5 weeks were maintained by the 4 four months assessment. CONCLUSION: Where postnatal mood difficulties are reported along with unsettled infant sleep difficulties, the evidence from this uncontrolled study, together with previously research, suggests that treatment which principally focuses on the baby-care issue can have a significant impact on many women's emotional health. PMID- 18511224 TI - Remarkably high inhibitory effects of docosahexaenoic acid incorporated into hybrid liposomes on the growth of tumor cells along with apoptosis. AB - Inhibitory effects of hybrid liposomes composed of l-alpha dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene (20) sorbitan monooleate (Tween 80) including polyunsaturated fatty acids or their ethyl esters (HL-PUFA) on the growth of human tumor cells were examined in vitro. Remarkably high inhibitory effects of HL including docosahexaenoic acid (HL-DHA) and alpha linolenic acid ethyl ester (HL-ALAE) on the growth of lung carcinoma (RERF-LC-OK and A549) cells, colon tumor (WiDr) cells and stomach tumor (MKN45) cells were obtained. The addition of vitamin E (alpha-tocopherol) to HL-DHA and -ALAE prevented almost completely the growth inhibition of A549 cells distinct from the other tumor cells used in this study. On the other hand, fluorescence microscopic and flow cytometric analyses indicated that the inhibitory effects of HL-DHA on the growth of RERF-LC-OK, WiDr and MKN45 cells could be attained through the induction of apoptosis. PMID- 18511225 TI - Sex determination from the radius and ulna in a modern South African sample. AB - With a large number of unidentified skeletal remains found in South Africa, the development of population specific osteometric standards is imperative. Forensic anthropologists need to have access to a variety of techniques to establish accurate demographic profiles from complete, fragmentary and/or commingled remains. No research has been done on the forearm of African samples, even though these bones have been shown to exhibit sexual dimorphism. The purpose of this paper is to develop discriminant function formulae to determine sex from the radius and ulna in a South African population. The sample consisted of 200 male and 200 female skeletons from the Pretoria Bone (University of Pretoria) and Raymond A. Dart (Witwatersrand University) collections. Sixteen standard anthropometric measurements were taken from the radius (9) and ulna (7) and subjected to stepwise and direct discriminant function analysis. Distal breadth, minimum mid-shaft diameter and maximum head diameter were the best discriminators of sex for the radius, while minimum mid-shaft diameter and olecranon breadth were selected for the ulna. Classification accuracy for the forearm ranged from 76 to 86%. The radius and ulna can be considered moderate discriminators for determining sex in a South African group. However, it is advised that these formulae are used in conjunction with additional methods to determine sex. PMID- 18511226 TI - Poly(ADP-ribose) polymerase activity in different pathologies--the link to inflammation and infarction. AB - DNA repair and aging are two phenomena closely connected to each other. The poly(ADP-ribosyl)ation reaction has been implicated in both of them. Poly(ADP ribose) was originally discovered as an enzymatic reaction product after DNA damage. Soon it became evident that it is necessary for regulation of different repair pathways. Also, evidence accumulated that poly(ADP-ribose) formation capacity is at least correlated with the life span of mammalian species. As a NAD(+)-consuming process, poly(ADP-ribosyl)ation can lead to cell death by energy depletion. This finding opened the area for investigation of poly(ADP-ribose) polymerase activity and polymer formation in pathologies. This review provides an introduction into the wide and complex field of poly(ADP-ribosyl)ation in different pathologies with regards of cell death regulation, inflammation and resulting tissue damage. PMID- 18511227 TI - Shoulder biomechanics. AB - The biomechanics of the glenohumeral joint depend on the interaction of both static and dynamic-stabilizing structures. Static stabilizers include the bony anatomy, negative intra-articular pressure, the glenoid labrum, and the glenohumeral ligaments along with the joint capsule. The dynamic-stabilizing structures include the rotator cuff muscles and the other muscular structures surrounding the shoulder joint. The combined effect of these stabilizers is to support the multiple degrees of motion within the glenohumeral joint. The goal of this article is to review how these structures interact to provide optimal stability and how failure of some of these mechanisms can lead to shoulder joint pathology. PMID- 18511228 TI - 18F-FDOPA PET/MRI fusion in patients with primary/recurrent gliomas: initial experience. AB - BACKGROUND AND PURPOSE: (18)F-FDOPA PET demonstrates higher sensitivity and specificity for gliomas than traditional [(18)F] FDG PET imaging. However, PET provides limited anatomic localization. The purpose of this study was to determine whether (18)F-FDOPA PET/MRI fusion can provide precise anatomic localization of abnormal tracer uptake and how this activity corresponds to MR signal abnormality. METHODS: Two groups of patients were analyzed. Group I consisted of 21 patients who underwent (18)F-FDOPA PET and MRI followed by craniotomy for tumor resection. Group II consisted of 70 patients with a pathological diagnosis of glioma that had (18)F-FDOPA PET and MRI but lacked additional pathologic follow-up. Fused (18)F-FDOPA PET and MRI images were analyzed for concordance and correlated with histopathologic data. RESULTS: Fusion technology facilitated precise anatomical localization of (18)F-FDOPA activity. In group I, all 21 cases showed pathology-confirmed tumor. Of these, (18)F-FDOPA scans were positive in 9/10 (90%) previously unresected tumors, and 11/11 (100%) of recurrent tumors. Of the 70 patients in group II, concordance between MRI and (18)F-FDOPA was found in 49/54 (90.1%) of patients with sufficient follow-up; in the remaining 16 patients concordance could not be determined due to lack of follow-up. (18)F-FDOPA labeling was comparable in both high- and low-grade gliomas and identified both enhancing and non-enhancing tumor equally well. In some cases, (18)F-FDOPA activity preceded tumor detection on MRI. CONCLUSION: (18)F-FDOPA PET/MRI fusion provides precise anatomic localization of tracer uptake and labels enhancing and non-enhancing tumor well. In a small minority of cases, (18)F-FDOPA activity may identify tumor not visible on MRI. PMID- 18511229 TI - Molecular structural characteristics as determinants of estrogen receptor selectivity. AB - Recent reports that a wide variety of natural and man-made compounds are capable of competing with natural hormones for estrogen receptors serve as timely examples of the need to advance screening techniques to support human health and ascertain ecological risk. Quantitative structure-activity relationships (QSARs) can potentially serve as screening tools to identify and prioritize untested compounds for further empirical evaluations. Computer-based QSAR molecular models have been used to describe ligand-receptor interactions and to predict chemical structures that possess desired pharmacological characteristics. These have recently included combined and differential relative binding affinities of potential estrogenic compounds at estrogen receptors (ER) alpha and beta. In the present study, artificial neural network (ANN) QSAR models were developed that were able to predict differential relative binding affinities of a series of structurally diverse compounds with estrogenic activity. The models were constructed with a dataset of 93 compounds and tested with an additional dataset of 30 independent compounds. High training correlations (r2=0.83-0.91) were observed while validation results for the external compounds were encouraging (r2=0.62-0.86). The models were used to identify structural features of phytoestrogens that are responsible for selective ligand binding to ERalpha and ERbeta. Numerous structural characteristics are required for complexation with receptors. In particular, size, shape and polarity of ligands, heterocyclic rings, lipophilicity, hydrogen bonding, presence of quaternary carbon atom, presence, position, length and configuration of a bulky side chain, were identified as the most significant structural features responsible for selective binding to ERalpha and ERbeta. PMID- 18511230 TI - Recovery and URICA stage-of-change scores in three marijuana treatment studies. AB - Data from three outpatient studies of treatment of marijuana dependence were used to test whether behavior change parallels stage movement in the transtheoretical model of change (TMC). This study examined one putative assumption of the TMC: individuals remaining in the preaction stages of change (i.e., precontemplation and contemplation) across time would show little change in their marijuana use. The University of Rhode Island Change Assessment (URICA) measure and two recent stage assignment algorithms were utilized to assess TMC stage status at baseline and end of treatment. The primary analyses employed paired-sample t tests of statistical equivalence and sign tests, and 34 out of the 36 tests demonstrated that individuals remaining in the preaction stages of change over the treatment period showed clinically important and statistically significant treatment outcomes. Clinicians and researchers should exercise caution in their use of URICA-based indices as meaningful markers of the recovery process. Better methods to measure stage status and test the TMC are needed if the model is to be used to guide substance abuse interventions. PMID- 18511231 TI - Selenocystine induces reactive oxygen species-mediated apoptosis in human cancer cells. AB - Epidemiological and clinical studies have demonstrated that dietary supplementation of selenium (Se) could reduce the incidence of human cancers. In this study, selenocystine, a nutritionally available selenoamino acid, was identified as a novel agent with broad-spectrum antitumor activity. A panel of eight human cancer cell lines was shown to be susceptible to selenocystine, with IC(50) values ranging from 3.6 to 37.0 microM. Selenocystine induced dose dependent apoptosis in A375, HepG2 and MCF7 cells was evaluated by flow cytometric analysis and annexin-V staining assay. Mechanistic studies showed time and dose-dependent increases in intracellular reactive oxygen species (ROS) in susceptible cancer cells (MCF7 and HepG2 cells) treated with selenocystine. However, selenocystine-induced ROS overproduction was not observed in non susceptible normal human fibroblast Hs68 cells. Significant DNA strand breaks were observed in selenocystine-treated MCF7 and HepG2 cells as examined by single cell gel electrophoresis (Comet assay). The thiol-reducing antioxidants, glutathione and N-acetylcysteine, inhibited intracellular ROS generation, DNA strand breaks and accumulation of sub-G1 population in MCF7 cells exposed to selenocystine. Our results suggest a possible role of ROS as a mediator of the signaling pathway of selenocystine-induced, DNA damage-mediated apoptosis in susceptible cancer cells. PMID- 18511232 TI - The stressor Criterion-A1 and PTSD: a matter of opinion? AB - Considerable controversy exists with regard to the interpretation and definition of the stressor "A1" criterion for Post Traumatic Stress Disorder (PTSD). At present, classifying an event as either traumatic (satisfying DSM-IV Criterion-A1 for PTSD), or non-traumatic (life event) is determined by the rater's subjective interpretation of the diagnostic criteria. This has implications in research and clinical practice. Utilizing a sample of 860 Australian adults, this study is the first to provide a detailed examination of the impact of event categorization on the prevalence of trauma and PTSD. Overall, events classified as non-traumatic were associated with higher rates of PTSD. Unanimous agreement between raters occurred for 683 (79.4%) events. As predicted, the categorization method employed (single rater, multiple rater-majority, multiple rater-unanimous) substantially altered the prevalence of Criterion-A1 events and PTSD, raising doubts about the functionality of PTSD diagnostic criteria. Factors impacting on the categorization process and suggestions for minimizing discrepancies in future research are discussed. PMID- 18511233 TI - NcDNAlign: plausible multiple alignments of non-protein-coding genomic sequences. AB - Genome-wide multiple sequence alignments (MSAs) are a necessary prerequisite for an increasingly diverse collection of comparative genomic approaches. Here we present a versatile method that generates high-quality MSAs for non-protein coding sequences. The NcDNAlign pipeline combines pairwise BLAST alignments to create initial MSAs, which are then locally improved and trimmed. The program is optimized for speed and hence is particulary well-suited to pilot studies. We demonstrate the practical use of NcDNAlign in three case studies: the search for ncRNAs in gammaproteobacteria and the analysis of conserved noncoding DNA in nematodes and teleost fish, in the latter case focusing on the fate of duplicated ultra-conserved regions. Compared to the currently widely used genome-wide alignment program TBA, our program results in a 20- to 30-fold reduction of CPU time necessary to generate gammaproteobacterial alignments. A showcase application of bacterial ncRNA prediction based on alignments of both algorithms results in similar sensitivity, false discovery rates, and up to 100 putatively novel ncRNA structures. Similar findings hold for our application of NcDNAlign to the identification of ultra-conserved regions in nematodes and teleosts. Both approaches yield conserved sequences of unknown function, result in novel evolutionary insights into conservation patterns among these genomes, and manifest the benefits of an efficient and reliable genome-wide alignment package. The software is available under the GNU Public License at http://www.bioinf.uni leipzig.de/Software/NcDNAlign/. PMID- 18511234 TI - Adherence-dependent shifts in the patterns of beta-amyloid peptides secreted by human mononuclear phagocytes. AB - Cells of the mononuclear phagocyte system are closely associated with vascular and neuritic beta-amyloid deposits in Alzheimer's disease. Using one-dimensional and newly developed two-dimensional Abeta-SDS-PAGE Western immunoblot techniques (1D/2D-Abeta-WIB) we investigated the patterns of Abeta peptides released by primary non-adherent and adherence-activated human mononuclear phagocytes in vitro. An overall increase of total released Abeta peptides (Abeta(total)) was observed in adherence-activated mononuclear phagocyte cultures. 2D-Abeta-WIB revealed that the proportion of Abeta(1-40) decreased significantly to 50.2+/ 5.4% (n=10) of Abeta(total) compared to 65.9+/-5.6% (n=7) in non-adherent cultures (p<0.0001, t=5.82). Abeta(1-42) accounted for only 3.0+/-2.1% of Abeta(total) and its proportion did not change significantly upon adherence (2.8+/-0.5% of Abeta(total)). In adherence-activated cultures we detected pronounced shifts in the fractional pattern of released Abeta peptides in favour of N-truncated species. The second most prominent Abeta peptide accounted for as much as 12.7+/-3.0% of Abeta(total) (2.0+/-1.2% in non-adherent cultures; p<0.0001, t=9.00) and was identified as Abeta(2-40) by comigration with a synthetic peptide and by N-terminal-specific antibodies. A strong increase of a further Abeta immunoreactive spot migrating at pI 5.45 was observed. It accounted for 9.2+/-1.7% of Abeta(total) as compared to 1.0+/-0.9% in non-adherent cultures (p<0.0001, t=11.61) and presumably represented a variant of Abeta(2-40) as determined by C-terminal Abeta(40)-specific immunoprecipitation and N-terminal specific immunodetection. Thus, mononuclear phagocytes might be one source of the N-truncated Abeta peptides regularly found in human plasma and are less likely to contribute substantially to plasma Abeta(1-42). PMID- 18511235 TI - B cell apotopes of the 60-kDa Ro/SSA and La/SSB autoantigens. AB - Apoptosis has been proposed to influence the initiation and diversification of autoimmunity to the Ro (SSA)/La (SSB) ribonucleoprotein (RNP) particle and serve as a target for autoantibody-mediated tissue injury. We have developed a new approach to B cell epitope mapping which identifies "apotopes," defined as epitopes expressed on the surface of apoptotic cells. Preliminary studies support a role for apotopes as diagnostic markers in systemic lupus erythematosus (SLE) and primary Sjogren's syndrome. For example, apotopes within the NH(2)-terminal and central regions of La react with the majority of sera from mothers of infants with congenital heart block. Furthermore, a Ro60 apotope is specific for a subset of SLE with isolated anti-Ro60 responses. The mapping of B cell apotopes may prove superior to standard epitope mapping by suggesting novel pathways of autoantibody production and identifying pathogenic species of autoantibodies. PMID- 18511236 TI - Systems genetics can provide new insights in to immune regulation and autoimmunity. AB - "Systems Genetics" detects variation in phenotypic traits and integrates this with underlying genetic variation. A powerful application of systems genetics is analyzing effects of genome-wide genetic variants on transcriptome-wide variation in gene expression. We see systems genetics as a new powerful technology which will empower research in genetics and in other disciplines. Here, we present preliminary analyses of systems genetics approaches to defining genetic interactions within the thymus, which is the key site for T lymphocyte development and imposition of immunological tolerance. PMID- 18511237 TI - Prevalence and significance of antibodies to citrullinated human papilloma virus 47 E2345-362 in rheumatoid arthritis. AB - It has been confirmed that antibodies to citrullinated profilaggrin306-324 may play important roles in RA. In this study, human papilloma virus (HPV)-47 E2345 362, homologous to profilaggrin306-324, was found using the NCBI BLAST program. Then, E2345-362 and citrullinated E2345-362, with arginine348 replaced by citrulline, were synthesized. The presence of antibodies against these peptides was examined by an enzyme-linked immunosorbent assay. Associations between these antibodies and the clinical and laboratory features of RA were evaluated. Although the prevalence and AU value of antibodies to the E2345-362 peptide were similar in RA and other rheumatic diseases, those of antibodies to the citrullinated E2345-362 peptide were significantly higher in RA than in other rheumatic diseases. Additionally, sera that were preincubated with cyclic citrullinated peptide (CCP) demonstrated lower AU values of anti-citrullinated E2345-362 peptide antibodies. Moreover, the prevalence of anti-CCP antibodies and that of anti-peptidylarginine deiminase (PADI4) antibodies in anti-citrullinated E2345-362-positive patients were all higher than those of anti-citrullinated E2345-362-negative patients. There were significant correlations between anti citrullinated E2345-362 and anti-PADI4. RA patients with antibodies to citrullinated E2345-362 had higher DAS28 scores, erythrocyte sedimentation rates, and radiographic progression than those without the antibodies. These results suggest that HPV-47 E2 may act as an autoantigen in RA. The increase in PADI4 may make it easier to citrullinate the HPV-47 E2345-362 peptide, leading to the subsequent immune responses. PMID- 18511238 TI - Risks and benefits of deliberate hypotension in anaesthesia: a systematic review. AB - This systematic review was performed to investigate and review the evidence on the risks and benefits of hypotensive anaesthesia in order to answer the following question: 'Should deliberate hypotension be used routinely during orthognathic surgery?' An electronic search on MEDLINE and the Cochrane Library database was carried out for all relevant articles using specific search keywords. All articles were classified by their levels of evidence. Studies with highest level of evidence and rated to have the lowest risk of bias were reviewed. Regarding the benefits of hypotensive anaesthesia, three studies reported significant decrease of blood loss in patients receiving hypotensive anaesthesia. Two studies reported a significant decrease in transfusion rate. Two studies demonstrated improved surgical field and significant reduction in operation time. In terms of risk, no significant changes in cerebral, cardiovascular, renal and hepatic functions in patients receiving hypotensive anaesthesia compared to control were reported. In conclusion, hypotensive anaesthesia appears to be effective in reducing blood loss. Serious consequences due to organ hypoperfusion are uncommon. Hypotensive anaesthesia can be justified as a routine procedure for orthognathic surgery especially bimaxillary osteotomy. Patient selection and appropriate monitoring are mandatory for this technique to be carried out safely. PMID- 18511239 TI - Further evidence for "hyper-priming" in thought-disordered schizophrenic patients using repeated masked category priming. AB - BACKGROUND: Previous research has yielded evidence for enhanced semantic priming in formal thought-disordered schizophrenia patients, a result that fits well with the hypothesis of disinhibited processes of spreading activation in this population. METHODS: The present study tested this hypothesis by using masked repetition priming, which yields reversed semantic priming effects in healthy participants. Assuming that performance in this paradigm relies on a balance between activation and inhibition processes in healthy participants, we compared formal thought-disordered schizophrenia patients, non-thought disordered schizophrenia patients, and healthy controls. RESULTS: For thought-disordered schizophrenia patients, we found a large positive semantic priming effect for dominant category exemplars (primed by the category name), whereas healthy controls had a small negative effect. For non-thought disordered patients, we found a non-significant, numerically positive effect. CONCLUSIONS: This result yields further evidence for the lack of inhibitory processes in thought disordered patients. PMID- 18511241 TI - Monitoring patients to improve physical health and treatment outcome. AB - Patients with schizophrenia suffer a number of comorbidities and a range of potential side effects from their medication, which contribute to a negative impact on their quality of life, and are sometimes responsible for lack of compliance and relapse. Side effects, in particular, can be identified by physicians, but only if they examine their patients regularly. Many physicians would suggest that this is good clinical practice for treating patients with schizophrenia, whatever the intended medication. This continuous monitoring might include weight gain, cognitive effects, and blood tests. In fact, patients appreciate being carefully monitored and it contributes to a good patient/physician relationship. This interaction has benefits in terms of impact on compliance and better control of symptoms due to treatment adherence (to appropriate doses), because it enables action before the patient stops taking medication. Good clinical practice suggests that investment in patient monitoring is offset by improved clinical outcomes. PMID- 18511240 TI - Genetically engineered mouse models for skin research: taking the next step. AB - Genetically engineered mouse models are invaluable to investigators in nearly all areas of biomedical research. The use of genetically engineered mice has allowed researchers to explore fundamental functions of genes in a mammal that shares substantial similarities with human physiology and pathology. Genetically engineered mice are often used as animal models of human diseases that are vital tools in investigating disease development and in developing and testing novel therapies. Gene targeting in embryonic stem cells allows endogenous genes to be specifically altered. As knowledge regarding precise genetic abnormalities underlying a variety of dermatological conditions continues to emerge, the ability to introduce corresponding alterations in endogenous gene loci in mice, often at a single base pair level, has become essential for detailed studies of these genetic diseases. In this review, we provide examples of mouse models harboring modified endogenous gene(s), generated using the technique commonly referred to as the "knock-in" approach, to exemplify the important and sometimes superior role of this methodology in dermatological research. PMID- 18511242 TI - Do we need another atypical antipsychotic? AB - Atypical antipsychotics were a great advance in the treatment of schizophrenia. But, there is still no atypical antipsychotic with an exceptional efficacy and safety profile for all patients. Clinicians are required to draw on their experiential knowledge to examine suitable options for individual patients. Following its suspension in 1998, the safety and efficacy of sertindole have been investigated in several post-marketing studies based in clinical settings. These have provided the safety data to support the reintroduction of sertindole, as well as specific examples demonstrating that certain patients, in particular, may benefit from a switch from other atypical antipsychotics to sertindole. Sertindole's individual and mostly favourable profile of treatment-emergent effects and safety allows for flexibility in treating patients. The propensity of sertindole to cause anticholinergic effects, which can be particularly troublesome, is small and, more recently, there have been suggestions that sertindole may have beneficial effects on cognition. PMID- 18511243 TI - A magnetic resonance imaging study of the entorhinal cortex in treatment resistant depression. AB - Despite a growing interest in this area, we continue to lack an understanding of the pathophysiology of depression and of treatment-resistant depression (TRD) in particular. The role of the medial temporal lobe, particularly the hippocampus, has been widely implicated in the aetiology of depression. However, related structures such as the entorhinal cortex have not been systematically examined. This research study aimed to examine possible abnormalities in the volume of the entorhinal cortex (ERC) in TRD patients. A group of 45 TRD patients and 30 healthy age- and sex-matched controls underwent magnetic resonance imaging (MRI). ERC volumes were manually traced from MRI data using ANALYZE software. An analysis of variance was conducted between subject groups and in the sexes separately while controlling for the effects of brain size via intracranial volume (ICV). Results revealed significant reductions in the volume of the left ERC of female patients. Although preliminary, our findings suggest that anatomical abnormalities in the ERC may confer vulnerability to treatment resistance. Confirmatory longitudinal studies are required to determine whether these abnormalities predate the onset of depression or are the result of a more chronic, treatment-resistant course of illness. PMID- 18511244 TI - [First orthodontic consultation: children or teenagers?]. AB - Orthodontic treatments aim at improving the positions of teeth, the form of dental arches and their reports relations. Determining the age to perform these treatments consists in defining the ideal moment to achieve them in the shortest time possible and with the best stability success. To know when to address the child or the teenager to the orthodontist, it is necessary for the pediatrician to observe the patient, his/her positions, orofacial functions (phonation, swallowing, aeration), his/her face and profile, oral cavity, particularly the way his/her teeth articulate in the three spatial directions. This orthodontic check-up allows the detection of abnormalities, for which two kinds of treatments exist. Firstly, early treatments which start in temporary dentition stage, in order to prevent or intercept abnormalities and, secondly, late treatments performed on permanent dentition, when well-developed bony and alveolar growths make it possible to correct these abnormalities. PMID- 18511246 TI - Glucomannan, a promising polysaccharide for biopharmaceutical purposes. AB - Over the last few decades, polysaccharides have gained increasing attention in the biomedical and drug delivery fields. Among them, glucomannan (GM), has become a particularly attractive polymer. In this paper, we review the physicochemical and biological properties which are decisive for the exploitation of GM as a biomaterial. These properties include the structural organization, molecular weight, solubility, viscosity, gelling properties and degradation behavior. Moreover, herein we analyze the possibilities of combining GM with other hydrophilic polymers, as well as the preparation of semisynthetic derivatives of GM, which may be of interest in the pharmaceutical context. Finally, we discuss the specific applications of GM in the drug delivery field. PMID- 18511247 TI - Studies on the reduction of tensile strength of tablets after roll compaction/dry granulation. AB - Roll compaction/dry granulation is a widely used technique for granulation. A major drawback is the reduction of tablet tensile strength compared to other granulation methods. The purpose of this study was to determine the reasons for the partial loss in compactibility. Microcrystalline cellulose of different particle sizes was roll-compacted/dry-granulated. The granules were sieved to obtain two sieve cuts and then compressed into tablets. The particle-size distribution within the sieve cut was determined using image analysis. The specific surface area of sieve cut was obtained by nitrogen adsorption. Heckel equation was used to determine the change in compressibility. The work-hardening phenomenon was found to be caused by a combination of particle-size enlargement and hardening of material. Although particle size of granules was equal, the use of smaller particles as raw material resulted in tablets with higher tensile strength due to higher specific surface area. Both work-hardening and particle size enlargement cause the partial loss in compactibility. The reduction in tensile strength could be compensated by producing smaller granules or using raw materials with small particle sizes. PMID- 18511248 TI - Evaluation of injection moulding as a pharmaceutical technology to produce matrix tablets. AB - The aim of this study was to develop sustained-release matrix tablets by means of injection moulding and to evaluate the influence of process temperature, matrix composition (EC and HPMC concentration) and viscosity grade of ethylcellulose (EC) and hydroxypropylmethylcellulose (HPMC) on processability and drug release. The drug release data were analyzed to get insight in the release kinetics and mechanism. Formulations containing metoprolol tartrate (30%, model drug), EC with dibutyl sebacate (matrix former and plasticizer) and hydrophilic polymer HPMC were extruded and subsequently injection moulded into tablets (375 mg, 10 mm diameter, convex-shaped) at temperatures ranging from 110 to 140 degrees C. Tablets containing 30% metoprolol and 70% ethylcellulose (EC 4mPa s) showed an incomplete drug release within 24 h (<50%). Increasing production temperatures resulted in a lower drug release rate. Substituting part of the EC fraction by HPMC (HPMC/EC-ratio: 20/50 and 35/35) resulted in faster and constant drug release rates. Formulations containing 50% HPMC had a complete and first-order drug release profile with drug release controlled via the combination of diffusion and swelling/erosion. Faster drug release rates were observed for higher viscosity grades of EC (Mw>20 mPa s) and HPMC (4000 and 10,000 mPa s). Tablet porosity was low (<4%). Differential scanning calorimetry (DSC) and X-ray powder diffraction studies (X-RD) showed that solid dispersions were formed during processing. Using thermogravimetrical analysis (TGA) and gel-permeation chromatography no degradation of drug and matrix polymer was observed. The surface morphology was investigated with the aid of scanning electron microscopy (SEM) showing an influence of the process temperature. Raman spectroscopy demonstrated that the drug is distributed in the entire matrix, however, some drug clusters were identified. PMID- 18511249 TI - 5-hydroxytryptamine (5HT)-induced valvulopathy: compositional valvular alterations are associated with 5HT2B receptor and 5HT transporter transcript changes in Sprague-Dawley rats. AB - Several drugs have been linked to valvulopathy in humans, including therapeutic agents for obesity, Parkinson's disease and migraine. There is increasing evidence that the 5-hydroxytryptamine 2B receptor (5HT2BR) activation and/or increased circulating 5HT (5-hydroxytryptamine) may play a significant role in the pathogenesis of drug-induced valvulopathy. In the present study, we investigated whether 7-day 5HT subcutaneous injections led to structural and compositional abnormalities in conjunction with transcriptomic modulation of 5HT2BR and 5HT transporter (5HTT) genes in the aortic and mitral valves of Sprague-Dawley (SD) rats. Subcutaneous injections of 5HT for 7 days resulted in thickening and compositional alteration of aortic and mitral valves in SD rats. More specifically, valve-leaflets from 5HT-treated rats had greater valve thickness, a higher amount of glycosaminoglycans (GAGs) and a lower amount of collagen. The compositional alteration was associated with up-regulation and down regulation of 5HT2BR and 5HTT genes, respectively. The present study strongly suggests that the activation of 5HT2BR and inhibition of 5HTT played a significant role in the pathogenesis of 5HT-induced valvulopathy in SD rats. Thus, these findings further highlight the necessity and/or utilization of animal models to screen potential valvular effects of serotonergic compounds. PMID- 18511250 TI - IL-5 and eosinophilia. AB - While Interleukin-5 (IL-5) is initially identified by its ability to support the growth and differentiation of activated B cells, overexpression of IL-5 significantly increases eosinophil numbers and antibody levels predominantly from an expanded population of B-1 cells in vivo. Conversely, mice lacking a functional gene for IL-5 or IL-5 receptor alpha chain (IL-5Ralpha) display a number of developmental and functional impairments in B cell and eosinophil lineages. In addition to the JAK-STAT and Btk pathway, the Ras-extracellular signal-regulated kinase (ERK) signals are important for IL-5-dependent cell survival. IL-5 critically regulates expression of genes involved in cell survival, IgH switch recombination, maturation in B cells and genes required for growth, survival, and effector function of eosinophils. IL-5Ralpha expression in B cells, but not in eosinophils is regulated by Oct-2. Eosinophilia is associated with a wide variety of conditions, including asthma and atopic diseases, helminth infections, drug hypersensitivity, and neoplastic disorders. In humans, the biologic effects of IL-5 are best characterized for eosinophils. The Sprouty related Ena/VASP homology 1-domain containing protein (Spred)-1 negatively controls eosinophil numbers and functions by modulating IL-5 signaling in allergic asthma. We will emphasize that IL-5 plays a pivotal role in the innate and acquired immune response and eosinophilia. PMID- 18511251 TI - The V-type H+-ATPase in vesicular trafficking: targeting, regulation and function. AB - Vacuolar-type H+-ATPase (V-ATPase)-driven proton pumping and organellar acidification is essential for vesicular trafficking along both the exocytotic and endocytotic pathways of eukaryotic cells. Deficient function of V-ATPase and defects of vesicular acidification have been recently recognized as important mechanisms in a variety of human diseases and are emerging as potential therapeutic targets. In the past few years, significant progress has been made in our understanding of function, regulation, and the cell biological role of V ATPase. Here, we will review these studies with emphasis on novel direct roles of V-ATPase in the regulation of vesicular trafficking events. PMID- 18511252 TI - From endocytosis to tumors through asymmetric cell division of stem cells. AB - Recent studies in vertebrate and invertebrate model organisms uncover the importance of endocytosis for biased signaling during asymmetric cell division. In stem cells, perturbing polarity and asymmetric division affect their selfrenewal causing exponential proliferation, thereby giving rise to cancer. An emerging pattern is that endocytosis controls asymmetric cell division, which underlies stem cell selfrenewal and defective selfrenewal is on the basis of tumorigenesis caused by cancer stem cells. PMID- 18511254 TI - High-speed particle detection in a micro-Coulter counter with two-dimensional adjustable aperture. AB - This article presents the fabrication and characterisation of a high-speed detection micro-Coulter counter with two-dimensional (2D) adjustable aperture and differential impedance detection. The developed device has been fabricated from biocompatible and transparent materials (polymer and glass) and uses the principle of hydrodynamic focusing in two dimensions. The use of a conductive solution for the sample flux and non-conductive solutions for the focalising fluxes provides an adjustable sample flow where particles are aligned and the resistive response concentrated, consequently enhancing the sensitivity and versatility of the device. High-speed counting of 20 microm polystyrene particles and 5 microm yeast cells with a rate of up to 1,000 particles/s has been demonstrated. Two-dimensional focusing conditions have been used in devices with physical cross-sectional areas of 180 microm x 65 microm and 100 microm x 43 microm, respectively, in which particles resulted undetectable in the absence of focusing. The 2D-focusing conditions have provided, in addition, increased detection sensitivity by a factor of 1.6 as compared to 1D-focusing conditions. PMID- 18511253 TI - MYSTs mark chromatin for chromosomal functions. AB - The MYST family of lysine acetyltransferases has been intensely studied because of its broad conservation and biological significance. In humans, there are multiple correlations between the enzymes and development and disease. In model organisms, genetic and biochemical studies have been particularly productive because of mechanistic insights they provide in defining substrate specificity, the complexes through which the enzymes function, and the sites of their activity within the genome. Established and emerging data from yeast reveal roles for the three MYST enzymes in diverse chromosomal functions. In particular, recent studies help explain how MYST complexes coordinate with other modifiers, the histone variant H2A.Z, and remodeling complexes to demarcate silent and active chromosomal domains, facilitate transcription, and enable repair of DNA damage. PMID- 18511255 TI - Analysis of the sensitivity and frequency characteristics of coplanar electrical cell-substrate impedance sensors. AB - A PDMS-glass based micro-device was designed and fabricated with 12 coplanar impedance sensors integrated for electrical cell-substrate impedance sensing (ECIS). The sensitivity and frequency characteristics of the sensors were investigated both theoretically (equivalent circuit model) and experimentally for the commonly used micro-electrode dimension scale (20-80 microm). The experimental results matched well with the theoretical model analysis and revealed that, within this micro-electrode dimension scale, as the electrode width decreased or as the total electrode length decreased the sensitivity of sensor increased over the whole sensing frequency range, whilst electrode to electrode distance had no influence on sensitivity. Through our frequency characteristics analysis, the whole frequency range could be divided into four parts. New functions describing the dominant components in each frequency range were defined and validated experimentally, and could be used to explain the phenomenon of an ECIS sensing frequency window. The contribution to the impedance measurement of cells growing on the edges of the electrodes was determined for the first time. Finally, novel proposals for ECIS sensor design and ECIS measurements were presented. PMID- 18511257 TI - Hemorrhagic stroke following elective cesarean delivery. AB - We present a case of hemorrhagic stroke after cesarean delivery under combined spinal-epidural anesthesia in an 35-year-old Hispanic patient treated with anticoagulants for protein C deficiency. She required vasopressor therapy for intraoperative hypotension and developed severe headache immediately after administration. To our knowledge, this is the first case of stroke occurring in a pregnant woman following vasopressor therapy for spinal anesthesia-induced hypotension. Although the exact cause of her hemorrhagic stroke is uncertain, the hypertensive response that may have led to the hemorrhagic stroke occurred following administration of commonly used doses of vasopressor agents. We discuss the possible causes of stroke. PMID- 18511258 TI - Right ventricular outflow tract tachycardia in the parturient. AB - Parturition is marked by physiological changes that may elicit electrocardiographic anomalies. Sustained right ventricular tachycardia is an uncommon arrhythmia that necessitates cardioversion using physical, pharmaceutical or electrical means. Patients with right ventricular tachycardia must be evaluated for right ventricular dysplasia. Long-term management of right ventricular tachycardia includes beta-adrenergic blockade and/or radio ablation of right ventricle outflow region usually below the pulmonary annuls. This report discusses the case of a 28-year-old previously healthy primigravid who experienced right ventricular outflow tract tachycardia during labor. PMID- 18511259 TI - Strategies to comply with the five-minute rule after maternal arrest. PMID- 18511260 TI - Stochastic neuronal cell fate choices. AB - Though many neuronal cell fate decisions result in reproducible outcomes, stochastic choices often lead to spatial randomization of cell subtypes. This is often the case in sensory systems where expression of a specific sensory receptor gene is selected randomly from a set of possible outcomes. Here, we describe recent findings elucidating the mechanisms controlling color photoreceptor subtypes in flies and olfactory receptor subtypes in worms and mice. Although well-known biological concepts such as lateral signaling and promoter selection play roles in these cases, fundamental questions concerning these choice mechanisms remain. PMID- 18511261 TI - The current excitement about copy-number variation: how it relates to gene duplications and protein families. AB - Following recent technological advances there has been an increasing interest in genome structural variants (SVs), in particular copy-number variants (CNVs)- large-scale duplications and deletions. Although not immediately evident, CNV surveys make a conceptual connection between the fields of population genetics and protein families, in particular with regard to the stability and expandability of families. The mechanisms giving rise to CNVs can be considered as fundamental processes underlying gene duplication and loss; duplicated genes being the results of 'successful' copies, fixed and maintained in the population. Conversely, many 'unsuccessful' duplicates remain in the genome as pseudogenes. Here, we survey studies on CNVs, highlighting issues related to protein families. In particular, CNVs tend to affect specific gene functional categories, such as those associated with environmental response, and are depleted in genes related to basic cellular processes. Furthermore, CNVs occur more often at the periphery of the protein interaction network. In comparison, protein families associated with successful and unsuccessful duplicates are associated with similar functional categories but are differentially placed in the interaction network. These trends are likely reflective of CNV formation biases and natural selection, both of which differentially influence distinct protein families. PMID- 18511262 TI - Anti-tumour activity of CS-7017, a selective peroxisome proliferator-activated receptor gamma agonist of thiazolidinedione class, in human tumour xenografts and a syngeneic tumour implant model. AB - The anti-tumour activity of the novel thiazolidinedione class peroxisome proliferator-activated receptor gamma (PPARgamma) agonist CS-7017 was investigated. CS-7017 activated PPARgamma-mediated luciferase expression with an EC(50) of 0.20 nM. In addition, CS-7017 was shown to be highly selective for PPARgamma amongst other PPAR subfamilies. CS-7017 inhibited the proliferation of the human anaplastic thyroid tumour cell line DRO and the pancreatic tumour cell line AsPC-1 in vitro at concentrations as low as 10 nM. In xenograft studies, CS 7017 inhibited the growth of the human colorectal tumour cell line HT-29 in nude mice as well as DRO in nude rats in a dose-dependent manner. At the same dose, an increase in the levels of adiponectin, a surrogate marker for PPARgamma activation, was also observed. CS-7017 prolonged the survival of mice inoculated with murine colorectal tumour Colon 38 with marginal tumour growth inhibition. These preclinical results support the potential utility of CS-7017 in a clinical setting. PMID- 18511263 TI - Studies on 5alpha-androst-16-en-3-one binding to porcine serum, plasma and testicular cytosolic fraction and to human serum. AB - The present study evaluated whether a specific androstenone-binding protein is present in porcine and human serum, and in the cytosolic fraction of porcine testis. The binding of [(3)H]-androstenone to serum and testicular cytosol was measured in the absence (total binding) and presence (non-specific binding) of unlabelled androstenone. The optimization of the assay is described. As a part of the assay validation, the binding of [(3)H]-dihydrotestosterone ([(3)H]-DHT) to porcine and human serum was also examined. As expected, specific binding of [(3)H]-DHT was detected in human serum, but not in porcine serum. No specific androstenone-binding protein was detected, either in porcine or human serum, or in the cytosolic fraction of porcine testis. The amount of non-specific binding of [(3)H]-androstenone was slightly lower in porcine serum compared to human serum. Between-animal variations in [(3)H]-androstenone binding were studied in plasma samples from 15 animals with androstenone concentrations ranging from 1.1 to 23.1 ng/mL. Mean values+/-standard deviations of binding in these samples were 15.2+/-0.9% for total binding and 15.9+/-0.8% for non-specific bindings. Low between-animal variations indicate that androstenone binding does not affect androstenone accumulation in fat. PMID- 18511264 TI - Bone marrow micrometastases in different solid tumors: pathogenesis and importance. AB - Early dissemination of cancer cells from the primary tumor via the circulatory system may result in the formation of microscopic metastatic deposits (micrometastases, MMs) in secondary compartments such as the bone marrow (BM), where there is a favorable environment for their subsequent growth and spread. MMs are considered the main reason for metastatic relapse in patients with early stage solid cancers after resection of the primary tumor. Although the molecular pathways leading to MMs remain only partly understood, there is increasing evidence that the detection of MMs in BM aspirates at the time of primary diagnosis is an independent prognostic factor, with a major influence in the stratification of these patients for adjuvant clinical treatment. Further potential applications of the detection of MMs include their use in monitoring therapeutic response or even in revealing targets for novel systemic therapies. All these intriguing possibilities are intensely investigated and carry great promise for radical improvements in the assessment and treatment of several epithelial cancers which are currently to blame for the majority of cancer related deaths in the industrialized world. PMID- 18511265 TI - Development of a new Cr(VI)-biosorbent from agricultural biowaste. AB - Among useless but abundant agricultural biowastes such as banana skin, green tea waste, oak leaf, walnut shell, peanut shell and rice husk, in this study, banana skin was screened as the most efficient biomaterial to remove toxic Cr(VI) from aqueous solution. X-ray photoelectron spectroscopy (XPS) study revealed that the mechanism of Cr(VI) biosorption by banana skin was its complete reduction into Cr(III) in both aqueous and solid phases and partial binding of the reduced Cr(III), in the range of pH 1.5-4 tested. One gram of banana skin could reduce 249.6 (+/-4.2)mg of Cr(VI) at initial pH 1.5. Namely, Cr(VI)-reducing capacity of banana skin was four times higher than that of a common chemical Cr(VI) reductant, FeSO(4).7H(2)O. To diminish undesirable/serious organic leaching from the biomaterial and to enhance removal efficiency of total Cr, its powder was immobilized within Ca-alginate bead. The developed Cr(VI)-biosorbent could completely reduce toxic Cr(VI) to less toxic Cr(III) and could remove almost of the reduced-Cr(III) from aqueous phase. On the basis of removal mechanisms of Cr(VI) and total Cr by the Cr(VI)-biosorbent, a kinetic model was derived and could be successfully used to predict their removal behaviors in aqueous phase. In conclusion, our Cr(VI)-biosorbent must be a potent candidate to substitute for chemical reductants as well as adsorbents for treating Cr(VI)-bearing wastewaters. PMID- 18511266 TI - Investigating the effects of anaerobic and aerobic post-treatment on quality and stability of organic fraction of municipal solid waste as soil amendment. AB - The use of OFMSW for biogas and compost production is considered as a sustainable strategy in saving valuable landfill space while producing valuable product for soil application. This study examines the effects of anaerobic and aerobic post treatment of OFMSW on the stability of anaerobic digestate and compost and soil quality using seed germination tests. Anaerobic digestion of OFMSW was carried out for fifteen days after which the residual anaerobic digestate was subjected to aerobic post-treatment for seventy days. Seed germination tests showed that fresh feedstock and digestates collected during anaerobic digestion and during the early stages of aerobic post-treatment were phytotoxic. However, phytotoxic effects were not observed in soils amended with the fully stabilised anaerobic digestate compost, ADC. It was also found that seed germination increases with dilution and incubation time, suggesting that lower soil application rates and longer lag periods between soil application of ADC and planting can reduce the amount of biodegradable organics in the ADC, thus enhancing the benefits of ADC as soil amendment. PMID- 18511267 TI - Effects of vibratory microscreening on proximate composition and recovery of poultry processing wastewater particulate matter. AB - Experiments were conducted to compare the effects of tertiary microscreen gap size on the proximate composition and rate of recovery of particulate matter from poultry processing wastewater (PPW). A high-speed vibratory screen was installed within the wastewater treatment area of a southeast US broiler slaughter plant after the existing primary and secondary mechanical rotary screens. Microscreen panels with nominal gap size openings of 212, 106 and 45mum were investigated. The particulate matter samples recovered were subjected to proximate analysis to determine percent moisture, fat, protein, crude fiber and ash. The average percent wet weight moisture (%WW) content for all samples was 79.1. The average percent dry matter (%DM) fat, protein, crude fiber and ash were 63.5, 17.5, 4.8 and 1.5, respectively. The mean concentration of total solids (TS) recovered from all microscreen runs was 668mg/L, which represents a potential additional daily offal recovery rate of 12.1metric tons (MT) per 3.78 million L (1.0 million gallons US) of PPW. There was no significant difference in the performance of the three microscreen gap sizes with regard to proximate composition or mass of particulate matter recovered. PMID- 18511268 TI - Characterization of products from the pyrolysis of rapeseed oil cake. AB - The main aim of this study was to investigate the composition of products from the pyrolysis of rapeseed oil cake in a fixed bed reactor at 400, 450, 500, 700 and 900 degrees C. The gas products mainly consisted of CO(2), CO, CH(4) and H(2)S at 500 degrees C. Empirical formula of bio-oil from the pyrolysis of rapeseed oil cake was CH(1.59)O(0.16)N(0.116)S(0.003) for 500 degrees C. Bio-oils mainly contained oleic acid, 1H-indole, 2,3,5-trimethoxy toluene, toluene, (Z)-9 octadecanamide, psoralene, phenol and phenol derivatives at all pyrolysis temperatures. Both non-aromatic and aromatic hydrocarbon compounds were determined in water phase of liquid product by Headspace-GC analysis. The heating values of bio-chars were found to be similar (24MJkg(-1)) at all pyrolysis temperatures. PMID- 18511269 TI - Use of response surface optimization for the production of biosurfactant from Rhodococcus spp. MTCC 2574. AB - The production of biosurfactant from Rhodococcus spp. MTCC 2574 was effectively enhanced by response surface methodology (RSM). Rhodococcus spp. MTCC 2574 was selected through screening of seven different Rhodococcus strains. The preliminary screening experiments (one-factor at a time) suggested that carbon source: mannitol, nitrogen source: yeast extract and meat peptone and inducer: n hexadecane are the critical medium components. The concentrations of these four media components were optimized by using central composite rotatable design (CCRD) of RSM. The adequately high R2 value (0.947) and F score 19.11 indicated the statistical significance of the model. The optimum medium composition for biosurfactant production was found to contain mannitol (1.6 g/L), yeast extract (6.92 g/L), meat peptone (19.65 g/L), n-hexadecane (63.8 g/L). The crude biosurfactant was obtained from methyl tert-butyl ether extraction. The yield of biosurfactant before and after optimization was 3.2 g/L of and 10.9 g/L, respectively. Thus, RSM has increased the yield of biosurfactant to 3.4-fold. The crude biosurfactant decreased the surface tension of water from 72 mN/m to 30.8 mN/m (at 120 mg L(-1)) and achieved a critical micelle concentration (CMC) value of 120 mg L(-1). PMID- 18511270 TI - An electrochemical sensor for detection of laccase activities from Penicillium simplicissimum in compost based on carbon nanotubes modified glassy carbon electrode. AB - An electrochemical sensor for detection of the activity of laccase from Penicillium simplicissimum isolated from the composting has been developed. The sensor is based on glassy carbon electrode modified with multi-wall carbon nanotubes (CNTs). The introduction of CNTs into this system can greatly enhance the electrochemical signal in this assay more sensitively, selectively and rapidly than that in conventional spectrophotometric assays. It was found that the optimal pH value of the electrolyte was 5.6. The results showed a good linear correlation between the current and the concentration of laccase activities measured by spectrophotometry, where the current slope was measured by chronoamperometry with a coefficient of 0.9835. Therefore, this electrochemical sensor can be used for rapid detection of laccase activity from P. simplicissimum. Furthermore, it may be potentially used for rapid quantification of P. simplicissimum according to the relationship between the laccase activities and the biomass. PMID- 18511271 TI - Biosorption of a reactive dye (Rhodamine-B) from an aqueous solution using dried biomass of activated sludge. AB - Low cost, locally available biomaterial was tested for its ability to remove reactive dyes from aqueous solution. Granules prepared from dried activated sludge (DAS) were utilized as a sorbent for the uptake of Rhodamine-B (Rh-B) dye. The effects of various experimental parameters (dye concentration, sludge concentrations, swelling, pretreatment and other factors) were investigated and optimal experimental conditions were ascertained. Nearly 15min was required for the equilibrium adsorption, and Rh-B dyes could be removed effectively. Dye removal performance of Rh-B and DAS increased with increasing concentrations. The acid pretreated biomass exhibited a slightly better biosorption capacity than alkali pretreated or non-pretreated biomass. The optimum swelling time for dye adsorption of the DAS within the swelling time range studied was 12h. Both the Freundlich and Langmuir isotherm models could describe the adsorption equilibrium of the reactive dye onto the activated sludge with the Langmuir isotherm showing the better agreement of the two. Second-order kinetic models confirmed the agreement. PMID- 18511272 TI - Adsorption and bioadsorption of granular activated carbon (GAC) for dissolved organic carbon (DOC) removal in wastewater. AB - In this study, the performances of GAC adsorption and GAC bioadsorption in terms of dissolved organic carbon (DOC) removal were investigated with synthetic biologically treated sewage effluent (BTSE), synthetic primary treated sewage effluent (PTSE), real BTSE and real PTSE. The main aims of this study are to verify and compare the efficiency of DOC removal by GAC (adsorption) and acclimatized GAC (bioadsorption). The results indicated that the performance of bioadsorption was significantly better than that of adsorption in all cases, showing the practical use of biological granular activated carbon (BGAC) in filtration process. The most significance was observed at a real PTSE with a GAC dose of 5g/L, having 54% and 96% of DOC removal by adsorption and bioadsorption, respectively. In addition, it was found that GAC adsorption equilibrium was successfully predicted by a hybrid Langmuir-Freundlich model whilst integrated linear driving force approximation (LDFA)+hybrid isotherm model could describe well the adsorption kinetics. Both adsorption isotherm and kinetic coefficients determined by these models will be useful to model the adsorption/bioadsorption process in DOC removal of BGAC filtration system. PMID- 18511273 TI - Bio-drying of municipal solid waste with high water content by aeration procedures regulation and inoculation. AB - To improve the water content reduction of municipal solid waste with high water content, the operations of supplementing a hydrolytic stage prior to aerobic degradation and inoculating the bio-drying products were conducted. A 'bio-drying index' was used to evaluate the bio-drying performance. For the aerobic processes, the inoculation accelerated organics degradation, enhanced the lignocelluloses degradation rate by 10.4%, and lowered water content by 7.0%. For the combined hydrolytic-aerobic processes, the inoculum addition had almost no positive effect on the bio-drying efficiency, but it enhanced the lignocelluloses degradation rate by 9.6% and strengthened the acidogenesis in the hydrolytic stage. Compared with the aerobic processes, the combined processes had a higher bio-drying index (4.20 for non-inoculated and 3.67 for the inoculated trials). Moreover, the lowest final water content occurred in the combined process without inoculation (50.5% decreased from an initial 72.0%). PMID- 18511274 TI - Decline in microbial activity does not necessarily indicate an end to biodegradation in MSW-biowaste: a case study. AB - The aim of this study was to examine whether a decline in microbial activity (i.e., CO(2) output) during the biodegradation of municipal solid waste (MSW) indicated an end to biodegradation and the appearance of a stable, final product. As the organic fraction of MSW was biodegraded in an 8 clamp Cambridge Batch System composter, CO(2) output declined by 50% and moisture content at the end of the process was <20%. Levels of biodegradable material remaining in the product were determined by the dynamic respiration index (DRI) method but despite 151 days in the composting system biodegradable material was still present at levels (24130mgO(2)/kgdry matter (DM)) which exceeded draft EU biowaste directive (2001) guidelines (10,000mgO(2)/kgDM). Further laboratory based incubations demonstrated that microbial activity and hence biodegradation of organic material could be restarted if moisture levels were adjusted suggesting that dehydration limited microbial activity. Low levels of microbial activity alone did not therefore indicate and end to biodegradation, biodegradable material was not exhausted and the final product was not stable which has serious implications for its end-use. PMID- 18511275 TI - Facile formation of hydrophilic derivatives of 5H-8,9-dimethoxy-5-[2-(N,N dimethylamino)ethyl]-2,3-methylenedioxydibenzo[c,h] [1,6]naphthyridin-6-one (ARC 111) and its 12-aza analog via quaternary ammonium intermediates. AB - Several new TOP1-targeting agents were prepared using as intermediates the N,N,N trimethyl quaternary ammonium salts of either ARC-111 or its 12-aza analog (ARC 31), 3 and 4, respectively. Direct displacement of the quaternary ammonium group with water, imidazole, alkylethylenediamines, or polyhydroxylated alkylamines provides a convenient means for furthering the structure-activity relationships associated with these non-camptothecin TOP1-targeting agents. PMID- 18511276 TI - Design, synthesis, and structure-activity relationships of piperidine and dehydropiperidine carboxylic acids as novel, potent dual PPARalpha/gamma agonists. AB - Several series of substituted dehydropiperidine and piperidine-4-carboxylic acid analogs have been designed and synthesized as novel, potent dual PPARalpha/gamma agonists. The SAR of these series of analogs is discussed. A rare double bond migration occurred during the basic hydrolysis of the alpha,beta-unsaturated dehydropiperidine esters 12, and the structures of the migration products were confirmed through a series of 2D NMR experiments. PMID- 18511277 TI - Discovery of benzamide tetrahydro-4H-carbazol-4-ones as novel small molecule inhibitors of Hsp90. AB - Hsp90 maintains the conformational stability of multiple proteins implicated in oncogenesis and has emerged as a target for chemotherapy. We report here the discovery of a novel small molecule scaffold that inhibits Hsp90. X-ray data show that the scaffold binds competitively at the ATP site on Hsp90. Cellular proliferation and client assays demonstrate that members of the series are able to inhibit Hsp90 at nanomolar concentrations. PMID- 18511278 TI - Discovery and initial SAR of arylsulfonylpiperazine inhibitors of 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1). AB - High-throughput screening of a small-molecule compound library resulted in the identification of a series of arylsulfonylpiperazines that are potent and selective inhibitors of human 11beta-Hydroxysteroid Dehydrogenase Type 1 (11beta HSD1). Optimization of the initial lead resulted in the discovery of compound (R) 45 (11beta-HSD1 IC(50)=3nM). PMID- 18511279 TI - Synthesis of new UV-B light absorbents: (Acetylphenyl)glycosides with antioxidant activities. AB - m-Acetylphenyl-beta-d-glucopyranosides and m-acetylphenyl-alpha/beta-d mannopyranosides were synthesized by the Koenigs-Knorr, Mitsunobu, and Helferich reactions as key glycosylation reactions, respectively. Their spectroscopic properties and antioxidative activities were characterized as potential ultraviolet B-ray absorbents. PMID- 18511280 TI - Pleuromutilin derivatives having a purine ring. Part 1: new compounds with promising antibacterial activity against resistant Gram-positive pathogens. AB - In the course of our research aimed at the discovery of metabolic stable pleuromutilin derivatives with more potent antibacterial activity against Gram positive pathogens than previous analogues, a series of compounds bearing a purine ring were prepared and evaluated. From SAR studies, we identified two promising compounds 85 and 87, which have excellent in vitro activity against a number of Gram-positive pathogens, including existing drug-resistant strains, and potent in vivo efficacy. PMID- 18511281 TI - Founder SH3TC2 mutations are responsible for a CMT4C French-Canadians cluster. AB - Charcot-Marie-Tooth polyneuropathies (CMT) are clinically and genetically heterogeneous. We describe a French-Canadian cluster of 17 recessive CMT cases belonging to 10 families with variable early-onset CMT and scoliosis. The patients demonstrate great intra- and inter-familial variability. Linkage analysis confirmed that all families are linked to CMT4C locus on chromosome 5q32 (multipoint LOD score of 9.06). Haplotype analysis suggests that two SH3TC2 mutations are present in this cohort. The majority of carrier chromosomes, 26 of 34 (76%), carry the c.2860C-->T mutation. Despite extensive sequencing, the other mutation is not yet uncovered. This study demonstrates that the clinical variability observed in CMT4C is due to other factors than the nature of the mutation and that further work is needed to better define the SH3TC2 gene to ensure the identification of all CMT4C mutations. PMID- 18511282 TI - Access to intensive care units: a survey in North-East Italy. AB - The factors associated with policies for allowing visitors into intensive care units (ICUs) are a debated issue in the nursing literature. The aim of this survey was to describe visiting policies in the ICUs of North-East Italy and to verify the hypothesis of an association between attitudes regarding accessibility to visitors and environmental, organisational or logistic variables. Data were collected by means of questionnaires sent by mail to head nurses of ICUs. The questionnaires were completed for 104 of the 110 ICUs contacted (94.5%). Visiting hours were generally less than 4h a day (86%) and only 14% of the ICUs reported imposing no restrictions. Children under 12 years old were rarely admitted (22%). Twenty-one percent of the ICUs reported always allowing exceptions, while 77% did so only under 'particular' circumstances. Visiting times were not associated with logistic and organisational factors, but rather with the type of ICU (p=0.000), city setting (p=0.009), exceptions to rules (p=0.029), allowing more than one person (p=0.016) and opening to children (p=0.001). Restrictive visiting policies emerged; paediatric units were generally more flexible. The association between the variables regarding visiting policy, such as visiting times and exceptions to rules, or allowing more than one person or children, seem to confirm how the rules are influenced mainly by the staff's attitude, which could be changed by continuing professional education. PMID- 18511283 TI - Synthesis and in vitro biological activity of retinyl retinoate, a novel hybrid retinoid derivative. AB - A new hybrid, retinyl retinoate 1, was synthesized with a condensing reaction between retinol and retinoic acid to improve the photo-stability, and the in vitro biological activity of the hybrid was analyzed. This retinol derivative had enhanced thermal stability and decreased photosensitivity, and exhibited decreased cell toxicity compared to that of retinol. In addition, RAR activity analysis showed that retinyl retinoate 1 had higher inhibitory activity against c Jun than retinol and showed superior effects on collagen synthesis compared to retinol. Thus, retinyl retinoate 1 may have the potential to be conveniently used as an additive in cosmetics for prevention and improvement of skin aging and medicines for the treatment of skin troubles due to its excellent stability under severe and accelerated conditions. PMID- 18511284 TI - 2-phenyl-4-piperazinylbenzimidazoles: orally active inhibitors of the gonadotropin releasing hormone (GnRH) receptor. AB - Antagonism of the gonadotropin releasing hormone (GnRH) receptor has shown positive clinical results in numerous reproductive tissue disorders such as endometriosis, prostate cancer and others. Traditional therapy has been limited to peptide agonists and antagonists. Recently, small molecule GnRH antagonists have emerged as potentially new treatments. This article describes the discovery of 2-phenyl-4-piperazinylbenzimidazoles as small molecule GnRH antagonists with nanomolar potency in in vitro binding and functional assays, excellent bioavailability (rat %F>70) and demonstrated oral activity in a rat model having shown significant serum leuteinizing hormone (LH) suppression. PMID- 18511285 TI - A soil sampling reference site: the challenge in defining reference material for sampling. AB - In the frame of the international SOILSAMP project, funded and coordinated by the Italian Environmental Protection Agency, an agricultural area was established as a reference site suitable for performing soil sampling inter-comparison exercises. The reference site was characterized for trace element content in soil, in terms of the spatial and temporal variability of their mass fraction. Considering that the behaviour of long-lived radionuclides in soil can be expected to be similar to that of some stable trace elements and that the distribution of these trace elements in soil can simulate the distribution of radionuclides, the reference site characterised in term of trace elements, can be also used to compare the soil sampling strategies developed for radionuclide investigations. PMID- 18511286 TI - Thorium series radionuclides in the environment: measurement, dose assessment and regulation. AB - Environmental radioactivity research is more advanced for the (238)U series than for the (232)Th series, for a number of reasons. One reason is that some radionuclides of the (232)Th series are more difficult to measure than the corresponding isotopes in the (238)U series, and measurement techniques are less widely available. The (232)Th series population doses, however, are sometimes significant. This paper analyses some main radionuclides in the (232)Th series in comparison with corresponding nuclides in the (238)U series, with regard to measurement techniques, dose coefficients and regulation needs. PMID- 18511287 TI - In situ gamma spectroscopy in environmental research and monitoring. AB - In situ gamma spectroscopy was introduced to determine the outdoor gamma dose rate from soil and to calculate the radionuclide concentration and the relative contribution to the dose rate. This paper reviews the most common and proven applications of in situ gamma spectroscopy-together with the most recent and innovative research outcomes obtained with this technique, particularly for its use indoors. Advantages and limitations of its utilization to assess environmental radioactivity-indoors and outdoors-are also discussed. PMID- 18511288 TI - Choice and criteria for selection of sampling strategies in environmental radioactivity monitoring. AB - Environmental radioactivity monitoring requires a sampling strategy to be defined, adopted and delivered using sound scientific principles. Statistical sampling delivers a set of sampling units from the population that is representative of all sampling units that could be taken. Such a representative set can then be used to draw inference(s) and conclusion(s) about the population based upon a statistical model. The environmental knowledge of the context in which the sampling is to be carried out plays a vital role in determining the appropriate statistical sampling strategy. PMID- 18511289 TI - Influence of natural radioactive aerosols on artificial radioactivity detection in the Spanish surveillance networks. AB - The device used for continuous measurements of artificial alpha and beta activity in Spanish radiological surveillance networks is the LB BAI 9850 monitor from the Berthold Company. The temporal variation of radon decay product equilibrium introduces a varying background signal in the artificial radioactivity in these monitors. This unwanted background signal can be significantly reduced by analyzing natural radioactive aerosols and their influence on the monitor. PMID- 18511290 TI - PHLiPPing the switch on Akt and protein kinase C signaling. AB - The Ser/Thr-specific phosphatase PHLPP [pleckstrin homology (PH) domain leucine rich repeat protein phosphatase] provides 'the brakes' for Akt and protein kinase C (PKC) signaling. The two isoforms of this recently discovered family, PHLPP1 and PHLPP2, control the amplitude and duration of signaling of Akt and PKC by catalyzing the dephosphorylation of the hydrophobic phosphorylation motif, a C terminal phosphorylation switch that controls these kinases. Aberrant regulation of either kinase accompanies many diseases, notably diabetes and cancer. By specifically dephosphorylating the hydrophobic motif, PHLPP controls the degree of agonist-evoked signaling by Akt and the cellular levels of PKC. This review focuses on the function of PHLPP1 and PHLPP2 in modulating signaling by Akt and PKC. PMID- 18511291 TI - Pro-inflammatory disequilibrium of the IL-1 beta/IL-1ra ratio in an experimental model of perinatal brain damages induced by lipopolysaccharide and hypoxia ischemia. AB - Bacterial infections and hypoxia/ischemia (H/I) are implicated in human neonatal brain damage leading to cerebral palsy (CP). We developed an animal model presenting similar perinatal brain damage by combining bacterial endotoxin and H/I insults. Interleukin (IL)-1beta is a mediator of brain damage and its action(s) is counteracted by its cognate anti-inflammatory IL-1 receptor antagonist (IL-1ra). We tested the hypothesis that the balance between agonist and antagonist in the IL-1 system is shifted towards inflammation in perinatal brains exposed to endotoxin and/or H/I. Lipopolysaccharide (LPS) and/or H/I enhanced both intracerebral IL-1beta mRNA and protein levels, with a maximum increase observed with combined LPS and H/I insults. Conversely, IL-1ra expression was significantly downregulated by LPS, H/I, or both combined, with a maximum magnitude of imbalance between IL-1beta and sIL-1ra noticed with the double hit. The nuclear factor (NF)kappaB component of the signaling pathway activated by IL-1beta-binding to its receptor was activated following exposure to LPS and/or H/I. We show for the first time that, perinatally, bacterial products, H/I, or both combined, induce downregulation in sIL-1ra expression concomitant with upregulation in IL-1beta. The resulting pro-inflammatory orientation in the IL-1/IL-1ra balance might play a role in the initiation of perinatal brain damages. PMID- 18511292 TI - Killing of human melanoma cells induced by activation of class I interferon regulated signaling pathways via MDA-7/IL-24. AB - Restoration of the tumor-suppression function by gene transfer of the melanoma differentiation-associated gene 7 (MDA7)/interleukin 24 (IL-24) successfully induces apoptosis in melanoma tumors in vivo. To address the molecular mechanisms involved, we previously revealed that MDA7/IL-24 treatment of melanoma cells down regulates interferon regulatory factor (IRF)-1 expression and concomitantly up regulates IRF-2 expression, which competes with the activity of IRF-1 and reverses the induction of IRF-1-regulated inducible nitric oxide synthase (iNOS). Interferons (IFNs) influence melanoma cell survival by modulating apoptosis. A class I IFN (IFN-alpha) has been approved for the treatment of advanced melanoma with some limited success. A class II IFN (IFN-gamma), on the other hand, supports melanoma cell survival, possibly through constitutive activation of iNOS expression. We therefore conducted this study to explore the molecular pathways of MDA7/IL-24 regulation of apoptosis via the intracellular induction of IFNs in melanoma. We hypothesized that the restoration of the MDA7/IL-24 axis leads to upregulation of class I IFNs and induction of the apoptotic cascade. We found that MDA7/IL-24 induces the secretion of endogenous IFN-beta, another class I IFN, leading to the arrest of melanoma cell growth and apoptosis. We also identified a series of apoptotic markers that play a role in this pathway, including the regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas-FasL. In summary, we described a novel pathway of MDA7/IL 24 regulation of apoptosis in melanoma tumors via endogenous IFN-beta induction followed by IRF regulation and TRAIL/FasL system activation. PMID- 18511293 TI - Endothelin receptor blockers protect against ischemia/reperfusion impairment of gastrointestinal motility in rats. AB - Intestinal ischemia/reperfusion (I/R) injury remains associated with high morbidity and mortality. The protective efficacy of the following endothelin (ET) receptor blockers: BQ-123 (ET(A) receptor), BQ-788 (ET(B)); tezosentan (dual ET blocker) was tested against the inhibition of gastrointestinal (GI) motility induced by intestinal I/R. Intestinal Evans blue transit was measured in untreated (UN) rats and animals subjected to skin incision (SI), I/R (1h superior mesenteric artery clamping followed by 2-24h reperfusion) or sham operation (SO). Surgical procedures were conducted under diethyl ether anesthesia. Anesthesia and SI did not affect the GI transit compared to UN rats. In contrast both SO and I/R significantly reduced GI motility, the latter evident at 2-24h of reperfusion. Tezosentan (1-10 mg/kg), BQ-123 and BQ-788 (0.1-1 mg/kg) protected against I/R induced inhibition of intestinal motility in a time- and dose-dependent manner at the early and late stages of reperfusion. Furthermore tezosentan alleviated the I/R-induced decrease in the contractile response of the longitudinal jejunal smooth muscle strips to carbachol in vitro. The serum ET(1-21) level was increased at 2h but not 24h of reperfusion compared to SO animals and ET(1-21) was higher in tezosentan pretreated rats. PMID- 18511294 TI - On the use of DHB/aniline and DHB/N,N-dimethylaniline matrices for improved detection of carbohydrates: automated identification of oligosaccharides and quantitative analysis of sialylated glycans by MALDI-TOF mass spectrometry. AB - This study demonstrates the application of 2,5-dihydrohybenzoic acid/aniline (DHB/An) and 2,5-dihydroxybenzoic acid/N,N-dimethylaniline (DHB/DMA) matrices for automated identification and quantitative analysis of native oligosaccharides by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Both matrices are shown to be superior to pure DHB for native glycans in terms of signal intensities of analytes and homogeneity of sample distribution throughout the crystal layer. On-target formation of stable aniline Schiff base derivatives of glycans in DHB/An and the complete absence of such products in the mass spectra acquired in DHB/DMA matrix provide a platform for automated identification of reducing oligosaccharides in the MALDI mass spectra of complex samples. The study also shows how enhanced sensitivity is achieved with the use of these matrices and how the homogeneity of deposited sample material may be exploited for quick and accurate quantitative analysis of native glycan mixtures containing neutral and sialylated oligosaccharides in the low-nanogram to mid picogram range. PMID- 18511295 TI - Identification of a caspase-derived N-terminal tau fragment in cellular and animal Alzheimer's disease models. AB - Biochemical modifications of tau proteins have been proposed to be among the earliest neurobiological changes in Alzheimer's disease (AD) and correlate better with cognitive symptoms than do beta-amyloid plaques. We have recently reported that adenovirus-mediated overexpression of the NH2 26-230aa tau fragment evokes a potent NMDA-mediated neurotoxic effect in primary neuronal cultures. In order to assess whether such N-terminal tau fragment(s) are indeed produced during apoptosis or neurodegeneration in vivo, we attempted to ascertain their presence in cell and animal models using an anti-tau antibody directed against the N terminal sequence of human protein located downstream of the caspase(s)-cleavage site DRKD(25)-QGGYTMHQDQ. We provide biochemical evidence that a caspase(s) cleaved NH2-terminal tau fragment of 20-22 kDa, consistent with the size of the NH2 26-230aa neurotoxic fragment of tau, is generated in vitro in differentiated human SH-SY5Y cells undergoing apoptosis by BDNF withdrawal or following treatment with staurosporine. In addition this NH2-terminally cleaved tau fragment, whose expression correlates with a significant up-regulation of caspase(s) activity, is also specifically detected in vivo in the hippocampus of 15 month-old AD11 transgenic mice, a model in which a progressive AD-like neurodegeneration is induced by the expression of transgenic anti-NGF antibodies. The results support the idea that aberrant activation of caspase(s), following apoptotic stimuli or neurodegeneration insults, may produce one or more toxic NH2 tau fragments, that further contribute to propagate and increase cellular dysfunctions in AD. PMID- 18511296 TI - Retinal TrkB receptors regulate neural development in the inner, but not outer, retina. AB - BDNF signaling through its TrkB receptor plays a pivotal role in activity dependent refinement of synaptic connectivity of retinal ganglion cells. Additionally, studies using TrkB knockout mice have suggested that BDNF/TrkB signaling is essential for the development of photoreceptors and for synaptic communication between photoreceptors and second order retinal neurons. Thus the action of BDNF on refinement of synaptic connectivity of retinal ganglion cells could be a direct effect in the inner retina, or it could be secondary to its proposed role in rod maturation and in the formation of rod to bipolar cell synaptic transmission. To address this matter we have conditionally eliminated TrkB within the retina. We find that rod function and synaptic transmission to bipolar cells is not compromised in these conditional knockout mice. Consistent with previous work, we find that inner retina neural development is regulated by retinal BDNF/TrkB signaling. Specifically we show here also that the complexity of neuronal processes of dopaminergic cells is reduced in conditional TrkB knockout mice. We conclude that retinal BDNF/TrkB signaling has its primary role in the development of inner retinal neuronal circuits, and that this action is not a secondary effect due to the loss of visual signaling in the outer retina. PMID- 18511297 TI - Ultrastructural matrix-mineral relationships in avian eggshell, and effects of osteopontin on calcite growth in vitro. AB - We investigated matrix-mineral relationships in the avian eggshell at the ultrastructural level using scanning and transmission electron microscopy combined with surface-etching techniques to selectively increase topography at the matrix-mineral interface. Moreover, we investigated the distribution of osteopontin (OPN) in the eggshell by colloidal-gold immunolabeling for OPN, and assessed the effects of this protein on calcite crystal growth in vitro. An extensive organic matrix network was observed within the calcitic structure of the eggshell that showed variable, region-specific organization including lamellar sheets of matrix, interconnected fine filamentous threads, thin film like surface coatings of proteins, granules, vesicles, and isolated proteins residing preferentially on internal {104} crystallographic faces of fractured eggshell calcite. With the exception of the vesicles and granules, these matrix structures all were immunolabeled for OPN, as were occluded proteins on the {104} calcite faces. OPN inhibited calcite growth in vitro at the {104} crystallographic faces producing altered crystal morphology and circular growth step topography at the crystal surface resembling spherical voids in mineral continuity prominent in the palisades region of the eggshell. In conclusion, calcite-occluded and interfacial proteins such as OPN likely regulate eggshell growth by inhibiting calcite growth at specific crystallographic faces and compartmental boundaries to create a biomineralized architecture whose structure provides for the properties and functions of the eggshell. PMID- 18511298 TI - Muscle activation patterns of knee flexors and extensors during passive and active movement of the spastic lower limb in chronic stroke patients. AB - The aim of this study was to describe the characteristics of spasticity, quantified as muscle activity during stretch, during passive and active movement. For this cross sectional study 19 stroke patients with spasticity in the lower limb were recruited. Reflex activity was studied with surface electromyography of knee flexor and extensor muscles during passive and active movement of the lower leg. On both the affected and unaffected side, root mean square values of the knee extensor muscles, while stretched, were higher during active than during passive movement (p<0.05). For the vastus lateralis (VL) the correlation was moderate (rho=0.54, p=0.022), for the rectus femoris (RF) high (rho=0.83, p<0.001). For the semitendinosus (ST) the correlation was low (rho=0.27) and not significant. During active movement the correlation between VL activity and activity of the antagonist ST, as an indicator for co-contraction of the affected muscles, was marked (rho=0.73, p=0.001). A moderate negative correlation was found between reflex activity of RF during passive stretch and the active range of motion (rho=-0.51, p=0.027). The results show that a passive stretch test alone is insufficient either as assessment method for spasticity during active motor tasks or as a measure for motor control. PMID- 18511299 TI - Stop the sadness: Neuroticism is associated with sustained medial prefrontal cortex response to emotional facial expressions. AB - Neuroticism is a personality trait associated with negative mood states, sensitivity to negative information, negative appraisal and vulnerability to psychopathology. Previous studies have associated the sustained processing of negative information (words) with individual differences such as rumination and depression but not with personality. In the current study, we aimed to investigate the relationship between neuroticism and changes in sustained patterns of activity within a brain region implicated in emotional self evaluation and appraisal, the Medial Prefrontal Cortex (MedPFC), when responding to emotional facial expressions (happy, fearful, and sad). We tested whether higher scores of neuroticism are associated with greater sustained patterns of brain activity in the MedPFC when responding to blocks of negative facial expressions. We found that higher scores of neuroticism were associated with greater sustained MedPFC activity throughout blocks of sad facial expressions, but not fearful or happy facial expressions. Based on the relationship between neuroticism and sensitivity to negative information, the current finding identifies a sustained temporal mechanism to this relationship. PMID- 18511301 TI - Time-course of "off-line" prefrontal rTMS effects--a PET study. AB - Low-frequency "off-line" repetitive transcranial magnetic stimulation (rTMS) over the course of several minutes has attained considerable attention as a research tool in cognitive neuroscience due to its ability to induce functional disruptions of brain areas. This disruptive rTMS effect is highly valuable for revealing a causal relationship between brain and behavior. However, its influence on remote interconnected areas and, more importantly, the duration of the induced neurophysiological effects, remain unknown. These aspects are critical for a study design in the context of cognitive neuroscience. In order to investigate these issues, 12 healthy male subjects underwent 8 H(2)(15)O positron emission tomography (PET) scans after application of long-train low-frequency rTMS to the right dorsolateral prefrontal cortex (DLPFC). Immediately after the stimulation train, regional cerebral blood flow (rCBF) increases were present under the stimulation site as well as in other prefrontal cortical areas, including the ventrolateral prefrontal cortex (VLPFC) ipsilateral to the stimulation site. The mean increases in rCBF returned to baseline within 9 min. The duration of this unilateral prefrontal rTMS effect on rCBF is of particular interest to those who aim to influence behavior in cognitive paradigms that use an "off-line" approach. PMID- 18511300 TI - The Brain Connectivity Workshops: moving the frontiers of computational systems neuroscience. AB - Understanding the link between neurobiology and cognition requires that neuroscience moves beyond mere structure-function correlations. An explicit systems perspective is needed in which putative mechanisms of how brain function is constrained by brain structure are mathematically formalized and made accessible for experimental investigation. Such a systems approach critically rests on a better understanding of brain connectivity in its various forms. Since 2002, frontier topics of connectivity and neural system analysis have been discussed in a multidisciplinary annual meeting, the Brain Connectivity Workshop (BCW), bringing together experimentalists and theorists from various fields. This article summarizes some of the main discussions at the two most recent workshops, 2006 at Sendai, Japan, and 2007 at Barcelona, Spain: (i) investigation of cortical micro- and macrocircuits, (ii) models of neural dynamics at multiple scales, (iii) analysis of "resting state" networks, and (iv) linking anatomical to functional connectivity. Finally, we outline some central challenges and research trajectories in computational systems neuroscience for the next years. PMID- 18511302 TI - The early context effect reflects activity in the temporo-prefrontal semantic system: evidence from electrical neuroimaging of abstract and concrete word reading. AB - Spatial and temporal characteristics of lexico-semantic retrieval are frequently examined with semantic context (i.e., priming) paradigms. These paradigms measure context (i.e., priming) effects in word processing evoked by semantically related context. Besides the well-known attention-dependent N400 context effect (>250 ms), recent studies demonstrate early automatic context effects in the P1-N1 time period (<200 ms). However, in visual word presentation the semantic origin of these early effects remains debated. This study examined spatio-temporal activation dynamics of the early context effect as well as the modulation of the effect by differences in structure and accessibility of verbal semantics existent in abstract and concrete words. The early context effect was measured in visually displayed words that followed semantically related single-word context. Spatial and temporal aspects of the effect were analyzed by applying topographic and source analyses on the word-triggered Event Related Potentials. The early context effect was enhanced in abstract compared to concrete words as indicated by a difference in the occurrence of P1-N1 transition map and a corresponding topographic dissimilarity (116-140 ms). This concreteness-dependent modulation demonstrates the sensitivity of the early context effect to structural differences in verbal semantics. Furthermore, the topographic difference was explained by enhanced activation in the left inferior prefrontal cortex for related compared to unrelated words in addition to temporo-parietal generators recruited in both conditions. The result suggests automatic feedforward processing of context-related information in temporo-prefrontal brain regions critical to semantic analysis. Taken together our findings show that the early context effect reflects activation processes in verbal semantic memory. PMID- 18511303 TI - In vivo PET study of 5HT(2A) serotonin and D(2) dopamine dysfunction in drug naive obsessive-compulsive disorder. AB - There are several lines of evidence, the majority indirect, suggesting that changes in serotonergic or dopaminergic neurotransmission may contribute to the pathogenesis of obsessive-compulsive disorder (OCD). We evaluated the co occurrence of serotonergic and dopaminergic dysfunctions in OCD subjects, all drug-naive, with no co-morbidity and homogeneous for symptoms. Each subject underwent two positron emission tomography (PET) scans to measure in vivo both serotonin (5-HT(2A)) and dopamine (D(2)) receptor distribution. For this, we used [11C]MDL and [11C]Raclopride, highly selective antagonists of 5-HT(2A) and D(2) receptors, respectively. The comparison with a control group was carried out using both voxel-wise (SPM2) and regions of interest (ROI) approaches. There was a significant reduction of 5-HT(2A) receptor availability in frontal polar, dorsolateral, and medial frontal cortex, as well as in parietal and temporal associative cortex of OCD patients. We also found a significant correlation between 5-HT(2A) receptor availability in orbitofrontal and dorsolateral frontal cortex and clinical severity, suggesting a specific role for serotonin in determining the OCD symptoms. There was also a significant reduction of [11C]Raclopride uptake in the whole striatum, particularly in the ventral portion, possibly reflecting endogenous dopaminergic hyperactivity. The co existence of serotonergic and dopaminergic dysfunction in the same homogeneous group of drug-naive OCD patients provides in vivo evidence for the complex molecular mechanisms of OCD, and represents the basis for further studies on the effect of therapeutic agents with specific modulatory effects on these neurotransmission systems. PMID- 18511304 TI - Self-paced movements induce high-frequency gamma oscillations in primary motor cortex. AB - There has been increasing interest in the functional role of high-frequency (>30 Hz) cortical oscillations accompanying various sensorimotor and cognitive tasks in humans. Similar "high gamma" activity has been observed in the motor cortex, although the role of this activity in motor control is unknown. Using whole-head MEG recordings combined with advanced source localization methods, we identified high-frequency (65 to 80 Hz) gamma oscillations in the primary motor cortex during self-paced movements of the upper and lower limbs. Brief bursts of gamma activity were localized to the contralateral precentral gyrus (MI) during self paced index finger abductions, elbow flexions and foot dorsiflexions. In comparison to lower frequency (10-30 Hz) sensorimotor rhythms that are bilaterally suppressed prior to and during movement (Jurkiewicz et al., 2006), high gamma activity increased only during movement, reaching maximal increase 100 to 250 ms following EMG onset, and was lateralized to contralateral MI, similar to findings from intracranial EEG studies. Peak frequency of gamma activity was significantly lower during foot dorsiflexion (67.4+/-5.2 Hz) than during finger abduction (75.3+/-4.4 Hz) and elbow flexion (73.9+/-3.7 Hz) although markedly similar for left and right movements of the same body part within subjects, suggesting activation of a common underlying network for gamma oscillations in the left and right motor cortex. These findings demonstrate that voluntary movements elicit high-frequency gamma oscillations in the primary motor cortex that are effector specific, and possibly reflect the activation of cortico subcortical networks involved in the feedback control of discrete movements. PMID- 18511306 TI - Increased cerebral perfusion in adult attention deficit hyperactivity disorder is normalised by stimulant treatment: a non-invasive MRI pilot study. AB - The neurobiological basis for attention deficit hyperactivity disorder (ADHD) has not yet been fully established, although there is a growing body of evidence pointing to functional and structural abnormalities involving the basal ganglia, cerebellum, and regions of frontal grey matter. The purpose of this study was to investigate regional cerebral perfusion in adults with ADHD and age-matched control subjects, and to assess the perfusion response to stimulant treatment in the ADHD group using a non-invasive magnetic resonance perfusion imaging technique. Whole-brain cerebral perfusion images were acquired from nine right handed male patients with ADHD and eleven age-matched control subjects using a continuous arterial spin labelling (CASL) technique. The ADHD group was assessed once on their normal treatment and once after withdrawing from treatment for at least one week. An automated voxel-based analysis was used to identify regions where the cerebral perfusion differed significantly between the ADHD and control groups, and where the perfusion altered significantly with stimulant treatment. Regional cerebral perfusion was increased in the ADHD group in the left caudate nucleus, frontal and parietal regions. Psychomotor stimulant treatment acted to normalise perfusion in frontal cortex and the caudate nucleus with additional decreases in parietal and parahippocampal regions. These findings highlight the potential sensitivity of non-invasive perfusion MRI techniques like CASL in the evaluation of perfusion differences due to illness and medication treatment, and provide further evidence that persistence of ADHD symptomatology into adulthood is accompanied by abnormalities in frontal and striatal brain regions. PMID- 18511305 TI - Functional neuroimaging correlates of finger-tapping task variations: an ALE meta analysis. AB - Finger-tapping tasks are one of the most common paradigms used to study the human motor system in functional neuroimaging studies. These tasks can vary both in the presence or absence of a pacing stimulus as well as in the complexity of the tapping task. A voxel-wise, coordinate-based meta-analysis was performed on 685 sets of activation foci in Talairach space gathered from 38 published studies employing finger-tapping tasks. Clusters of concordance were identified within the primary sensorimotor cortices, supplementary motor area, premotor cortex, inferior parietal cortices, basal ganglia, and anterior cerebellum. Subsequent analyses performed on subsets of the primary set of foci demonstrated that the use of a pacing stimulus resulted in a larger, more diverse network of concordance clusters, in comparison to varying the complexity of the tapping task. The majority of the additional concordance clusters occurred in regions involved in the temporal aspects of the tapping task, rather than its execution. Tapping tasks employing a visual pacing stimulus recruited a set of nodes distinct from the results observed in those tasks employing either an auditory or no pacing stimulus, suggesting differing cognitive networks when integrating visual or auditory pacing stimuli into simple motor tasks. The relatively uniform network of concordance clusters observed across the more complex finger-tapping tasks suggests that further complexity, beyond the use of multi-finger sequences or bimanual tasks, may be required to fully reveal those brain regions necessary to execute truly complex movements. PMID- 18511307 TI - MEG imaging of sensorimotor areas using inter-trial coherence in vibrotactile steady-state responses. AB - We utilized a novel analysis technique to identify brain areas that activate synchronously during the steady-state interval of responses to vibrotactile stimulation of the right index finger. The inter-trial coherence at the stimulation rate (23 Hz) was determined for whole-brain neural activity estimates based on a linearly-constrained minimum variance beamformer applied to the MEG data. Neural activity coherent with the stimulus occurred in the contralateral primary somatosensory cortex in all subjects, and matched well with equivalent dipole modeling of the same data. Subsets of subjects exhibited additional loci of strongly coherent activity in the contralateral primary motor cortex, posterior parietal cortex, and supplementary motor area, as well as in deeper brain structures above the brainstem. An activation delay of 7 ms from deep structures to cortical areas was estimated based on the mean phase at each coherent neural source within a single subject. This new approach - volumetric mapping of the statistical parameter of inter-trial coherence in steady-state oscillations - broadens the range of MEG beamformer applications specifically for identifying brain areas that are synchronized to repetitive stimuli. PMID- 18511308 TI - The effect of incongruity and instability on contact stress directional gradients in human cadaveric ankles. AB - OBJECTIVE: Measure incongruity and instability-associated changes in transient contact stress directional gradients in a human cadaveric ankle model. METHODS: Seven cadaveric ankles were subjected to quasi-physiologic forces and motion under intact conditions and with a stepoff incongruity of the anterior one-third of the distal tibia. Anterior/posterior forces were modulated to create incongruous specimens that either maintained a stable articulation between the talus and distal tibia or developed gross instability during motion. Real-time contact stresses were measured using a custom-designed ankle stress transducer at 132 Hz. Contact stress data were differentiated using a central-differencing formula to calculate transient contact stress directional gradients over the entire ankle articulation. RESULTS: Transient 95th percentile contact stress directional gradient values increased by 30 and 100%, respectively, in stable incongruous and unstable-incongruous conditions compared to intact conditions. Compared to stable-incongruous conditions, transient contact stress directional gradients increased by 60% in unstable-incongruous conditions. CONCLUSIONS: Instability resulted in greater percentage increases in transient contact stress directional gradients compared to incongruity. Pathologic increases in contact stress directional gradients potentially play an important role in the etiology of post-traumatic arthritis. PMID- 18511309 TI - Emotional and learning behaviour in mice overexpressing heat shock protein 70. AB - The effects of inducible heat shock protein 70 (HSP70) on emotional and learning behaviour as well as hippocampal long-term potentiation was investigated in transgenic HSP70 overexpressing mice. In active two-way avoidance learning (shuttle box) as well as spatial 8-arm radial maze learning, the HSP70 overexpressing mice showed diminished learning performance. In several tests there was no indication of differences in anxiety behaviour between transgenic mice and wild-type mice. This suggests that impairment in learning behaviour is unrelated to the learning task and motivational aspects of behaviour. To investigate the neurophysiological correlate of learning, long-term potentiation experiments were performed. In transversal hippocampal slices, an enhanced amplitude of the population spike was found in HSP70 overexpressing mice. It was hypothesised that enhanced potentiation in conjunction with potentiation effects due to learning led to learning impairment. PMID- 18511310 TI - Molecular analysis of the digestive microbiota in a gnotobiotic mouse model during antibiotic treatment: Influence of Saccharomyces boulardii. AB - The probiotic Saccharomyces boulardii is a non-pathogenic yeast that has been proven efficient in the prevention of antimicrobial-associated diarrhea and of Clostridium difficile associated colitis. We evaluated the influence of the administration of S. boulardii on the composition of the fecal microbiota in a human microbiota-associated mouse model. This evaluation was run before, during and after a 7-day oral treatment with amoxicillin clavulanic acid. Predominant groups of bacteria were quantified with fluorescence in situ hybridization combined with flow cytometry using group-specific 16S rRNA targeted oligonucleotide probes designed for the Eubacteria, Bacteroides-Porphyromonas Prevotella, Clostridium coccoides-Eubacterium rectale, Faecalibacterium prausnitzii, Clostridium histolyticum, Lactobacillus-Enterococcus and Enterobacteriaceae groups and Bifidobacterium species. S. boulardii did not quantitatively alter the total anaerobic microbiota nor the dominant bacterial groups. During the antibiotic treatment in the two groups of mice receiving the yeast or not, the level of Enterobacteriaceae and Bacteroides groups increased when the C. coccoides-E. rectale group decreased dramatically. After the antibiotic treatment was discontinued, the return to the initial level was reached more rapidly in the S. boulardii-treated mice than in the control mice (p<0.05) for the C. coccoides-E. rectale and Bacteroides-Porphyromonas-Prevotella groups. This quicker recovery of normal intestinal microbiota equilibrium after antibiotic therapy could be a mechanism for S. boulardii preventive effect on antibiotic-associated diarrhea in humans. PMID- 18511311 TI - The stem cell marker CD133 meets the endosomal compartment--new insights into the cell division of hematopoietic stem cells. PMID- 18511312 TI - The hemolysis kinetics of psoriatic red blood cells. AB - Psoriasis has been reported to be associated with several red blood cell (RBC) membrane alterations including: a membrane fluidity decrease, a significant elevation of Na(+)-K(+)and quantitative changes of erythrocyte membrane proteins that may indicate red cell cytoskeleton impairment. The aim of the present study was to analyse the hemolytic behaviour of psoriatic RBCs. The osmotic behaviour of RBCs was examined by analysing the kinetics of hemolysis. The method is based on the measurement of the transmitted light (lambda=700 nm) scattered by a suspension of red blood cells subjected to osmotic stress in the stopped-flow regime. The transmittance as a function of time, which describes the lysis kinetics, can be satisfactorily fitted with a mathematical model which assumes three cell populations in each sample: cells that do not lyse in the experimental conditions and cells that undergo fast and slow lysis. A comparison of the erythrocyte hemolytic kinetics of blood samples from psoriatic patients and healthy subjects showed distinct differences. The fraction of hemolyzed erythrocytes for control samples was about 20%; for psoriatic ones 12.6% (P<0.001). In control blood samples the fraction of fast-breaking cells was greater (about 61% of lysed cells) than in psoriatic ones (about 56%). The parameter T(fast), describing the time of fast kinetics and T(slow), which reflects the rupturing time of cells belonging to the fraction of slow hemolysing cells were significantly higher for psoriatic erythrocytes than for control cells (P<0.001). It was shown that the psoriatic erythrocyte has a low propensity for hemolysis and that its plasma membrane is distinctly more resistant to osmotic stress, probably related to decreased bilayer fluidity and low cell deformability. The results of this study showed that the kinetics of hemolysis may be a promising method for detecting erythrocytes with defective plasma membrane components and/or defective cytoskeleton organization and indirectly can provide some information on cell function. PMID- 18511313 TI - Prevalence of gastric ulcers in Thoroughbred broodmares in pasture: a preliminary report. AB - Gastroscopic examinations were performed in 62 Thoroughbred broodmares (33 pregnant, 29 non-pregnant) at one breeding farm to investigate the prevalence of gastric ulceration. Age, pregnancy status, race earnings, last race start, herd size, medical history, number of live foals, breeding years, feed type and number of feedings were recorded, plus coat condition and body condition score were determined. Twenty-one mares were re-evaluated after foaling, and the foaling date, foal weight at birth and placenta weight were recorded. The overall prevalence of gastric ulcers was 70.9%, with a median ulcer score of 3.0 (range: 2-5). Most ulcers were present on the squamous portion of the stomach, while two mares had glandular ulcers. There were no differences in the presence, location and severity of gastric ulcers between pregnant and non-pregnant mares. Furthermore, there were no significant associations between the variables measured and the presence of gastric ulceration. The prevalence of gastric ulceration in this specific population of horses was higher than expected and further investigation is warranted to determine the factors that contributed to this finding. PMID- 18511314 TI - Long-lived states in solution NMR: theoretical examples in three- and four-spin systems. AB - Long-lived spin states have been observed in a variety of systems. Although the dynamics underlying the long lifetimes of these states are well understood in the case of two-spin systems, the corresponding dynamics in systems containing more spins appear to be more complex. Recently it has been shown that a selection rule for transitions mediated by intramolecular dipolar relaxation may play a role in determining the lifetimes of long-lived states in systems containing arbitrary numbers of spins. Here we present a theory of long-lived states in systems containing three and four spins and demonstrate how it can be used to identify states that have little or no intramolecular dipolar relaxation. PMID- 18511315 TI - Lipoprotein lipase (LPL) is highly expressed and active in the ovary of European sea bass (Dicentrarchus labrax L.), during gonadal development. AB - The oocytes of many fish species accumulate high amounts of neutral lipids as a caloric reserve for embryonic and larval development. We propose that lipoprotein lipase (LPL, EC 3.1.1.34) plays an important role in supplying the oocytes with fatty acids and we have cloned its cDNA from the ovary of sea bass, and determined the patterns of LPL activity and LPL mRNA expression in the ovary. The cDNA obtained was 3051 bp long with an open reading frame encoding 518 amino acids. The amino acid sequence has a high similarity and shows similar structural features to LPL of other species. Northern blot analysis revealed LPL expression in adipose tissue and gonads only. LPL activity and LPL mRNA expression in the ovary was very high in fish with a gonadosomatic index (GSI) above 5, coinciding with the appearance of a high number of lipid droplets in the ooplasm. The LPL mRNA expression was localised to the follicle cells surrounding the oocyte. Our results suggest that LPL is likely to play an important role in the incorporation of neutral lipids into the oocytes, and that follicle cells, in addition to participating in steroidogenesis, also may be important in building up oocyte lipid reserves. PMID- 18511316 TI - Short-term thermoregulatory adjustments in a South American anseriform, the black necked swan (Cygnus melanocoryphus). AB - Avian bioenergetic studies suggest that, compared with other vertebrates, birds are efficient thermoregulators. However, most avian physiological studies have been performed in species of small body masses (less than 1 kg). In contrast to what might be anticipated, thermoregulatory abilities of large, flying birds are scarcely studied, especially in temperate zones and aquatic systems. In order to determine short-term metabolic adjustment after thermal challenge, we studied the bioenergetics of a South American anseriform, the black-necked swan (Cygnus melanocoryphus). Our results suggest that this swan species exhibits lower resting metabolic rate compared with other anseriforms, and some hetherothermia. In addition, the black-necked swans in our study changed "wet" thermal conductance at different ambient temperatures. At our working Ta range (5, 10, 15, 20 and 25 degrees C) calculated values were considerably higher than expected (23%, 26%, 39% and 51% higher than expected, respectively). Our results differ considerably from the only two previous reports in swan species, suggesting that C. melanocoryphus, perhaps due to its temperate distribution, is more sensitive to changes in environmental temperature. PMID- 18511317 TI - Cellular uptake of sepiapterin and push-pull accumulation of tetrahydrobiopterin. AB - Cellular uptake of sepiapterin resulted in an efficient accumulation of tetrahydrobiopterin. Tetrahydrobiopterin is much less permeable across the cell membrane than sepiapterin or dihydrobiopterin, the precursors of the tetrahydrobiopterin-salvage pathway. The uptake of sepiapterin by the cell was examined under metabolic arrest with N-acetylserotonin, an inhibitor of sepiapterin reductase. The release profile of previously accumulated sepiapterin was also analyzed. Two routes were clearly distinguishable, namely rapid and slow. Both were apparently bi-directional and equilibrating in type. Each route was connected to non-mixable pools somehow separated in the cell. The rapid process was too fast to analyze by the current methods of cell handling. The slower process was associated with conversion of sepiapterin to tetrahydrobiopterin in the absence of N-acetylserotonin, suggesting that this route opens into the cytosolic compartment where use of the salvage pathway was strongly driven by sepiapterin reductase and dihydrofolate reductase with a supply of NADPH which favors tetrahydrobiopterin accumulation. Consequently, sepiapterin was enforcedly taken up by the cell where it accumulated tetrahydrobiopterin in the cytosol in continuous manner. PMID- 18511318 TI - A novel single-base substitution (c.1124A>G) that activates a 5-base upstream cryptic splice donor site within exon 11 in the human mitochondrial acetoacetyl CoA thiolase gene. AB - Most mutations related to aberrant splicing occur in conserved splice acceptor and donor sites. Some exonic mutations also affect splicing. We identified and characterized a point mutation (c.1124A>G) in an Australian patient (GK43) with mitochondrial acetoacetyl-CoA thiolase (T2) deficiency. GK43 is a homozygote of c.1124A>G, which activates a cryptic splice donor site 5 bases upstream from c.1124A>G within exon 11, causing aberrant splicing in most transcripts. The aberrant splicing results in c.1120-1163 (44-base) deletion, causing a frameshift in T2 mRNA. A mini-gene splicing experiment confirmed that the c.1124A>G substitution was responsible for this aberrant splicing. This cryptic splice site has a Shapiro and Senapathy score (70.0) in a normal sequence but if mutated, the score (84.3) becomes higher than the one in the authentic splice donor site of intron 11 (81.4). This is an example in which a point mutation activates a cryptic splice donor site motif that is used preferentially over a downstream authentic splice site. PMID- 18511319 TI - Heparin cofactor II-thrombin complex: a biomarker of MPS disease. AB - The mucopolysaccharidoses are a group of lysosomal storage disorders caused by defects in the degradation of glycosaminoglycans. Each disorder is characterized by progressive multi-system disease with considerable clinical heterogeneity. The clinical heterogeneity of these disorders is thought to be related to the degree of the metabolic block in glycosaminoglycan degradation which in turn is related to the underlying mutation at the respective locus. There are currently no objective means other than longitudinal clinical observation, or the detection of a recurrent genetic mutation to accurately predict the clinical course for an individual patient, particularly when diagnosed early. In addition, there are no specific disease biomarkers that reflect the total body burden of disease. The lack of specific biomarkers has made monitoring treatment responses and predicting disease course difficult in these disorders. The recent introduction of enzyme replacement therapy for MPS I, II, and VI highlights the need for objective measures of disease burden and disease responsiveness. We show that serum levels of heparin cofactor II-thrombin complex is a reliable biomarker of the mucopolysaccharidoses. Untreated patients have serum levels that range from 3 to 112-fold above control values. In a series of patients with varying severity of mucopolysaccharidosis I, the serum complex concentration was reflective of disease severity. In addition, serum heparin cofactor II-thrombin levels showed responsiveness to various treatment regimens. We propose that serum levels of heparin cofactor II-thrombin complex may provide an important assessment and monitoring tool for patients with mucopolysaccharidosis. PMID- 18511320 TI - Interference of iohexol with radioiodine thyroid uptake in the hyperthyroid cat. AB - Absorbed thyroid dose and effective half-life were determined in 46 hyperthyroid cats after treatment with a low dose (mean 111MBq) of radioiodine intravenously. Thirteen of these cats had received iohexol for glomerular filtration rate (GFR) measurement within 24h before treatment with radioiodine in view of another ongoing study at our institution. Pre-therapy values were obtained for total thyroxine (TT(4)) and for the thyroid to salivary gland ratio with sodium pertechnetate gamma-camera imaging. All cats underwent post-therapy scans at 24, 48 and 120 h for evaluation of radioactive iodine uptake (RAIU) and the effective half-life of radioiodine. The absorbed dose was calculated from the cumulative activity with Olinda software. Both groups were comparable in age, TT(4) and the ratio of thyroid activity to salivary gland activity. Statistical analysis revealed a significant decreased absorbed dose in the thyroid in the iohexol group. This decreased uptake was not accompanied by an decreased effective half life of the radioiodine. The variation of inter-individual RAIU decreased in this group and more homogenous absorbed doses were obtained. No significant difference in outcome could be demonstrated. However, a tendency towards a higher number of residual hyperthyroidism in the iohexol group was noted (15 versus 6% in control group). This study demonstrates that iohexol interferes with the uptake of radioiodine in the hyperthyroid cat but does not provoke increased turnover. In this study, albeit including a small number of cats, outcome did not seem to be significantly affected. PMID- 18511321 TI - Molecular epidemiology and risk factors of bloodstream infections caused by extended-spectrum beta-lactamase-producing Klebsiella pneumoniae A case-control study. AB - OBJECTIVES: To study the prevalence, risk factors, outcome, and molecular epidemiology in patients with bacteremia caused by extended-spectrum beta lactamase (ESBL)-producing Klebsiella pneumoniae (Kp) (cases), in comparison with patients with bacteremia caused by a susceptible Kp (controls). METHODS: This was a retrospective case-control study including all episodes of Kp bacteremia for the period 1993 to 2002 at a referral hospital for adults in Mexico. ESBL production was tested for by E-test. All isolates were typed by pulsed field gel electrophoresis (PFGE). A subset of isolates underwent plasmid analysis, conjugal transfer of cefotaxime resistance to Escherichia coli J53-2, isoelectric focusing bioassay, colony-blot hybridization, PCR, and sequencing. RESULTS: Of the 121 patients with bacteremia due to Kp included in the study, 17 (14.0%) had an ESBL Kp isolate (cases). Multivariate analysis identified prior use of cephalosporins (OR 7.6, 95% CI 1.1-53.5; p=0.039) and stay in the intensive care unit (ICU; OR 5.6, 95% CI 1.1-27.9; p=0.033) as significant risk factors. No differences were observed in hospital stay or mortality after the event. Multi-drug resistance was more frequent in ESBL-Kp. There was no clonal predominance. A distinct beta lactamase profile was identified, which included a combination of TEM-1 (pI 5.4) and SHV-5 (pI 8.2) in 13/17 ESBL-Kp isolates. Cefotaxime resistance was transferred by conjugation in 14/17 isolates with a >120-kb plasmid encoding ESBL. CONCLUSIONS: The prevalence of ESBL-Kp was found to be lower than that previously reported in Latin America. ESBL-Kp bacteremia was not associated with a worse clinical outcome. We were able to identify a plasmid-mediated horizontal dissemination over the 10-year period. PMID- 18511322 TI - Validation of the Accutrend lactate meter for hyperlactatemia screening during antiretroviral therapy in a resource-poor setting. AB - BACKGROUND: The use of highly active antiretroviral therapy (HAART) results in multiple side effects that may jeopardize the life of the patient being treated with antiretroviral drugs. In resource-poor settings it is difficult to definitively diagnose lactic acidosis by laboratory measurement of lactate. Point of-care (POC) devices are helpful in the measurement of lactate levels and have been validated in the intensive care unit setting, but not in a busy outpatient clinic. The objective of this study was to assess the sensitivity and specificity of the Accutrend lactate meter in the diagnosis of hyperlactatemia/lactic acidosis in patients on nucleoside reverse transcriptase inhibitor (NRTI) containing regimens (stavudine). DESIGN AND METHODS: This was a cross-sectional study of 120 patients on HAART (lamivudine, stavudine, efavirenz, or nevirapine) with symptoms of/or recovering from hyperlactatemia/lactic acidosis. Simultaneous testing of the same blood sample was undertaken on the Accutrend handheld lactate analyzer and a reference instrument (Beckman CX7 Synchron machine). A venous lactate level <2.2mmol/l was considered as normal. RESULTS: The mean lactate value obtained from the Accutrend meter was 2.89mmol/l and from the reference instrument was 2.78mmol/l. The standard deviation for Accutrend meter was 1.14mmol/l vs.1.42mmol/l for the Beckman instrument. The sensitivity obtained for the Accutrend meter was 95.9% (95% CI 87.7-98.9%) and the specificity 63.8% (95% CI 48.5-76.9%). The positive predictive value was 80.5% (95% CI 70.3-87.9%) and the negative predictive value was 90.9% (95% CI 74.5-97.6%). CONCLUSIONS: The Accutrend lactate meter is an appropriate device for screening patients on HAART with symptoms of hyperlactatemia/lactic acidosis. The use of this device decreases analytic and intervention time, preventing further morbidity and mortality in patients on an NRTI (stavudine)-based regimen. PMID- 18511323 TI - Cerebellar ataxia following prolonged use of metronidazole: case report and literature review. AB - Cerebellar toxicity is a rare adverse event in patients treated with metronidazole. Here, we present a patient who developed cerebellar toxicity accompanied by objective abnormalities on magnetic resonance imaging, and review the literature on this unusual reaction. Discontinuation of metronidazole almost always results in resolution of symptoms and structural lesions. PMID- 18511324 TI - Type 2 diabetes: a well-characterised but suboptimally controlled disease. Can we bridge the divide? AB - From a pathophysiological point of view, type 2 diabetes is a well-characterised disease, since the glycaemic disorders result from three main mechanisms (the De Fronzo's triumvirate): a defect of beta-cell function, decreased disposal of glucose in peripheral tissues and overproduction of glucose by the liver. Each defect is subject to 24-h circadian variations and to inevitable worsening with time. As a consequence, therapeutic strategies should reflect whether patients retain sufficient insulin secretion or suffer from a more severe secretory defect that progresses from being responsive to oral diabetic agents to the insulin requiring stage. Identifying the different pathophysiological stages is a prerequisite for successful therapeutic strategies. This assessment can be done by considering on the one hand the HbA(1c) and on the other the glycaemic profiles. For the latter, either discontinuous (self-monitoring of blood glucose) or continuous glucose monitoring can be used. However, many difficulties remain for bridging the divide between well-understood pathophysiological concepts and suboptimal glycaemic control achieved in clinical practice. The main drawback is the difficulty in providing therapies at recommended doses to stochastic phenomena such as either intestinal absorption of carbohydrates or fluctuations in both pharmacokinetics and pharmacodynamics of hypoglycaemic agents. PMID- 18511325 TI - [Merkel cell carcinoma: outcome and role of radiotherapy]. AB - Merkel cell carcinoma (MCC) are rare neuroendocrine malignant tumor of the skin, occurring in elderly patients. It affects primarily the sun-exposed areas of the skin, with approximately 50% of all tumors occurring in the face and neck and 40% in the extremities. Immunohistochemical markers (CK20+, CK7- and TTF1-) are used to distinguish between MCC and other tumors. MCC have a tendency to rapid local progression, frequent spread to regional lymph nodes and distant metastases. Due to the rarity of the disease, the optimal treatment has not been fully defined. Localized stages (stages I and II) are treated by surgical excision of the primary tumor (with 2 to 3 cm margin) and lymphadenectomy in case of node positive disease, followed by external beam radiotherapy (EBRT) to a total dose of 50 to 60Gy in the tumor bed. Adjuvant EBRT has been shown to decrease markedly locoregional recurrences and to increase survival in recent studies. Treatment of lymph nodes area is more controversial. Chemotherapy is recommended only for metastatic disease. PMID- 18511326 TI - Risk analysis and implants. AB - According to the Medical Devices Directive, both the preparation for clinical trials and marketing of implants require that a risk analysis is performed. This paper presents a risk analysis for a dental implant in the framework of the risk management process carried out for the preparation of a multi-centre clinical trial, where likely hazards, failure modes and their severities, probabilities and detectabilities are assessed, together with a review of the related scientific literature. The clinical study aimed to evaluate a new ion implantation-based implant surface designed for the promotion of more extensive and faster osseointegration. PMID- 18511327 TI - A new mutation in the GCH1 gene presents as early-onset Parkinsonism. PMID- 18511328 TI - Socioeconomic inequalities in mortality in Barcelona: a study based on census tracts (MEDEA Project). AB - The aim of this study is to describe inequalities in socioeconomic indicators and in mortality by sex in the census tracts of Barcelona city during the period 1996 2003. The results show that there is excess mortality in coastal and northern areas. This distribution is similar to that of socioeconomic deprivation and therefore there is an association between mortality and socioeconomic indicators, not only for total mortality but also for the specific causes of death studied. This type of analysis can be useful for planning of public health policy since it allows small areas with high mortality risk to be detected. PMID- 18511329 TI - From acute musculoskeletal pain to chronic widespread pain and fibromyalgia: application of pain neurophysiology in manual therapy practice. AB - During the past decade, scientific research has provided new insight into the development from an acute, localised musculoskeletal disorder towards chronic widespread pain/fibromyalgia (FM). Chronic widespread pain/FM is characterised by sensitisation of central pain pathways. An in-depth review of basic and clinical research was performed to design a theoretical framework for manual therapy in these patients. It is explained that manual therapy might be able to influence the process of chronicity in three different ways. (I) In order to prevent chronicity in (sub)acute musculoskeletal disorders, it seems crucial to limit the time course of afferent stimulation of peripheral nociceptors. (II) In the case of chronic widespread pain and established sensitisation of central pain pathways, relatively minor injuries/trauma at any locations are likely to sustain the process of central sensitisation and should be treated appropriately with manual therapy accounting for the decreased sensory threshold. Inappropriate pain beliefs should be addressed and exercise interventions should account for the process of central sensitisation. (III) However, manual therapists ignoring the processes involved in the development and maintenance of chronic widespread pain/FM may cause more harm then benefit to the patient by triggering or sustaining central sensitisation. PMID- 18511330 TI - Simple anatomical information improves the accuracy of locating specific spinous processes during manual examination of the low back. AB - The objective of the study was to test whether a teaching protocol including simple anatomical information on the surface anatomy of spinous processes, improves physiotherapy students' ability to accurately locate selected thoracic and lumbar spinal segments - T12 and L3. First year physiotherapy students were allocated to Group 1 (n=35) and Group 2 (n=34). Both groups were taught to identify spinous processes by counting up from the sacrum, but Group 2 received supplementary anatomical information on the shapes and vertical length of the tips of L5 to T12 spinous processes. The spinous processes of L3 and T12 were located by two experienced physiotherapists and marked on a model using an invisible skin marker. Volunteer students were asked to locate these spinous processes and accuracy was confirmed using an ultraviolet lamp. Students with supplementary anatomical information (Group 2) were significantly better at locating T12 (difference in proportions 36% (95% confidence interval 14 to 51%)) and both T12 and L3 (difference in proportions 33% (11 to 48%)). Group 2 students were also better than Group 1 students at locating L3 (difference in proportions 28% (4 to 48%)), but the difference was not significant. Including simple anatomical information when teaching manual examination skills improved the accuracy of locating specific low back spinal levels. PMID- 18511331 TI - The TNF receptor and Ig superfamily members form an integrated signaling circuit controlling dendritic cell homeostasis. AB - Dendritic cells (DC) constitute the most potent antigen presenting cells of the immune system, playing a key role bridging innate and adaptive immune responses. Specialized DC subsets differ depending on their origin, tissue location and the influence of trophic factors, the latter remain to be fully understood. Myeloid associated lymphotoxin-beta receptor (LTbetaR) signaling is required for the local proliferation of lymphoid tissue DC. This review focuses on the LTbetaR signaling cascade as a crucial positive trophic signal in the homeostasis of DC subsets. The noncanonical coreceptor pathway comprised of the immunoglobulin (Ig) superfamily member, B and T lymphocyte attenuator (BTLA) and TNFR superfamily member, herpesvirus entry mediator (HVEM) counter regulates the trophic signaling by LTbetaR. Together both pathways form an integrated signaling circuit achieving homeostasis of DC subsets. PMID- 18511332 TI - Direct three-dimensional myocardial strain tensor quantification and tracking using zHARP. AB - Images of myocardial strain can be used to diagnose heart disease, plan and monitor treatment, and to learn about cardiac structure and function. Three dimensional (3D) strain is typically quantified using many magnetic resonance (MR) images obtained in two or three orthogonal planes. Problems with this approach include long scan times, image misregistration, and through-plane motion. This article presents a novel method for calculating cardiac 3D strain using a stack of two or more images acquired in only one orientation. The zHARP pulse sequence encodes in-plane motion using MR tagging and out-of-plane motion using phase encoding, and has been previously shown to be capable of computing 3D displacement within a single image plane. Here, data from two adjacent image planes are combined to yield a 3D strain tensor at each pixel; stacks of zHARP images can be used to derive stacked arrays of 3D strain tensors without imaging multiple orientations and without numerical interpolation. The performance and accuracy of the method is demonstrated in vitro on a phantom and in vivo in four healthy adult human subjects. PMID- 18511333 TI - Groupwise surface correspondence by optimization: representation and regularization. AB - Groupwise optimization of correspondence across a set of unlabelled examples of shapes or images is a well-established technique that has been shown to produce quantitatively better models than other approaches. However, the computational cost of the optimization is high, leading to long convergence times. In this paper, we show how topologically non-trivial shapes can be mapped to regular grids, hence represented in terms of vector-valued functions defined on these grids (the shape image representation). This leads to an initial reduction in computational complexity. We also consider the question of regularization, and show that by borrowing ideas from image registration, it is possible to build a non-parametric, fluid regularizer for shapes, without losing the computational gain made by the use of shape images. We show that this non-parametric regularization leads to a further considerable gain, when compared to parametric regularization methods. Quantitative evaluation is performed on biological datasets, and shown to yield a substantial decrease in convergence time, with no loss of model quality. PMID- 18511334 TI - Oomycete RXLR effectors: delivery, functional redundancy and durable disease resistance. AB - To manipulate host defences, plant pathogenic oomycetes secrete and translocate RXLR effectors into plant cells. Recent reports have indicated that RXLR effectors are translocated from the extrahaustorial matrix during the biotrophic phase of infection and that they are able to suppress PAMP-triggered immunity. Oomycete genomes contain potentially hundreds of highly diverse RXLR effector genes, providing the potential for considerable functional redundancy and the consequent ability to readily shed effectors that are recognised by plant surveillance systems without compromising pathogenic fitness. Understanding how these effectors are translocated, their precise roles in virulence, and the extent to which functional redundancy exists in oomycete RXLR effector complements, are major challenges for the coming years. PMID- 18511335 TI - Studies of the spin Hamiltonian parameters and local structure for ZnO:Cu2+. AB - The spin Hamiltonian parameters (the g factors and the hyperfine structure constants) and local structure for ZnO:Cu2+ are theoretically studied from the perturbation formulas of these parameters for a 3d9 ion under trigonally distorted tetrahedra. The ligand orbital and spin-orbit coupling contributions are taken into account from the cluster approach due to the significant covalency of the [CuO4](6-) cluster. According to the investigations, the impurity Cu2+ is suggested not to locate on the ideal Zn2+ site in ZnO but to undergo a slight outward displacement (approximately 0.01 angstroms) away from the ligand triangle along C3 axis. The calculated spin Hamiltonian parameters are in good agreement with the observed values. The validity of the above impurity displacement is also discussed. PMID- 18511336 TI - Synthesis of a novel fluorescent probe and investigation on its interaction with nucleic acid and analytical application. AB - A novel fluorescent probe N-(N-(2-(4-morpholinyl)ethyl)-4-acridinecarboxamide) alpha-alanine (N-(N-(ME)-4-ACA)-alpha-ALA) was synthesized. The structure was characterized by 1H NMR, MS, elemental analysis, fluorescent and ultraviolet spectra. This new compound exhibited high binding affinity to DNA, intense fluorescence and high water solubility. Experiment indicated that the fluorescent intensity was quenched when DNA was added. A method for DNA determination based on the quenching fluorescence (lambda(ex)=258nm, lambda(em)=451nm) of N-(N-(ME)-4 ACA)-alpha-ALA was established. Under optimal conditions (pH 7.2, CN-(N-(ME)-4 ACA)-alpha-ALA)=3 x 10(-6) mol L(-1)), the linear range is 0.1-4.0 microg mL(-1) for both fish semen (fsDNA) and calf thymus DNA (ct-DNA). The corresponding determination limits are 4.6 ng mL(-1) for fsDNA and 5.1 ng mL(-1) for ct-DNA, respectively. The relative standard deviation is 1.0%. Thus this compound can be used as a DNA fluorescent probe. The experiments proved that the interaction mode between N-(N-(ME)-4-ACA)-alpha-ALA and DNA was groove binding. The modified Rosenthal's graphical method gave the binding constant of 1.0 x 10(6) L mol(-1) and a binding size of 0.31 base pairs per bound drug molecule. PMID- 18511337 TI - EPR and optical study of Mn2+ doped ammonium tartrate single crystals. AB - EPR study of Mn2+ doped ammonium tartrate single crystals is carried out at room temperature. The spin Hamiltonian parameters are: gx=1.9225+/-0.0002, gy=1.9554+/ 0.0002, gz=2.1258+/-0.0002, A=(78+/-2) x 10(-4) cm(-1), B=(75+/-2) x 10(-4) cm( 1), D=(191+/-2) x 10(-4) cm(-1), E=(61+/-2) x 10(-4) cm(-1) and a=(22+/-1) x 10( 4) cm(-1) for site I and gx=1.9235+/-0.0002, gy=1.9574+/-0.0002, gz=2.0664+/ 0.0002, A=(78+/-2) x 10(-4) cm(-1), B=(75+/-2) x 10(-4) cm(-1), D=(180+/-2) x 10( 4) cm(-1), E=(57+/-2) x 10(-4) cm(-1) and a=(22+/-1) x 10(-4) cm(-1) for site II, respectively. The observed optical bands are fitted with inter-electronic repulsion parameters (B and C), crystal field parameter (Dq) and Trees correction (alpha) and the values found are B=752, C=2438, Dq=765 and alpha=76 cm(-1). The data obtained are further used to discuss the surrounding crystal field and the nature of metal-ligand bonding in the crystal. PMID- 18511338 TI - Spectroscopic and electrochemical properties of iron(II) complexes of polydentate Schiff bases containing pyrazine, pyridine and imidazole groups. AB - We report the synthesis and spectroscopic/electrochemical properties of iron(II) complexes of polydentate Schiff bases generated from 2-acetylpyridine and 1,3 diaminopropane, acetylpyrazine and 1,3-diaminopropane, and from 2-acetylpyridine and L-histidine. The complexes exhibit bis(diimine)-iron(II) chromophores in association with pyrazine, pyridine or imidazole groups displaying contrasting pi acceptor properties. In spite of their open geometry, their properties are much closer to those of macrocyclic tetraimineiron(II) complexes. An electrochemical/spectroscopic correlation between E degrees (FeIII/II) and the energies of the lowest MLCT band has been observed, reflecting the stabilization of the HOMO levels as a consequence of the increasing backbonding effects in the series of compounds. Mossbauer data have also confirmed the similarities in their electronic structure, as deduced from the spectroscopic and theoretical data. PMID- 18511339 TI - Characterization of temperate phage Che12 and construction of a new tool for diagnosis of tuberculosis. AB - A temperate phage, Che12, able to infect Mycobacterium tuberculosis, was isolated from soil samples taken from tuberculosis sanatorium area in Chennai, India. The plaque morphology of this phage showed varying grades of turbidity on lawns of M. tuberculosis. The temperate nature of Che12 was established by super infection immunity. Phage integration into the host genomic DNA was confirmed by Southern hybridization using Che12 DNA as a probe. PCR amplification and sequencing of a part of the integrated phage genome in a M. tuberculosis lysogen also confirmed the temperate nature of Che12. The morphology of the phage particles was observed by electron microscopy, revealing similarities to other mycobacteriophages like L5, D29 and TM4. A luciferase reporter phage, phAETRC16, was constructed by cloning firefly luciferase gene into Che12. Infection of viable M. tuberculosis cells by phAETRC16 resulted in expression of luciferase leading to sustained light output. Che12, a true temperate phage infecting M. tuberculosis, is thus ideally suited for developing a diagnostic tool facilitating rapid diagnosis of M. tuberculosis. PMID- 18511340 TI - International Workshop on Tuberculosis Vaccines: May 13-17, 2007, Varadero, Matanzas, Cuba. PMID- 18511341 TI - Two international forums. Editorial. PMID- 18511342 TI - ESFH congress held in Dusseldorf, Germany September 16-19, 2008. PMID- 18511343 TI - Patients with type 2 diabetes undergoing coronary artery bypass graft surgery: predictors of outcomes. AB - Type 2 diabetic (T2DM) patients undergoing CABG have worse outcomes than non diabetic patients, especially if insulin-treated. The purpose of this study was to determine if pre-hospital management of diabetes (including insulin) and risk factors affect outcomes. METHODS: T2DM patients were recruited at time of CABG and followed 3 months. Variables were illness level factors: insulin treatment, risk factor management, heart disease measures; Predisposing Factors: demographics, health status (SF-36), Geriatric Depression Scale (GDS), Charlson Co-morbidity Index (CCI); OUTCOMES: post-operative length of stay (PLOS), complications, re-hospitalisation. RESULTS: 317 T2DM patients (mean age 66, female 37%) had poor health status, met half the risk factor goals, and 27% had depressive symptoms. Mean PLOS was 7.4+/-5 days, 44% had complications, and 32% were re-hospitalised. Insulin-treated patients were at higher risk and had worse outcomes. Independent predictors of outcomes: CCI and measures of left ventricular function (all outcomes); risk factor management (complications); insulin treatment, GDS, age (PLOS); female sex, BMI, PLOS (re-hospitalisation). CONCLUSIONS: Insulin treatment in T2DM patients indicates patients at high risk of poor outcomes. Intensive management and rehabilitation is needed to improve outcomes, and patients would benefit from optimal management of risk factors pre- and post-CABG. PMID- 18511344 TI - The number and function of circulating dendritic cells may limit effector memory CD4+ T-cell responses in HIV patients responding to antiretroviral therapy. AB - Some HIV patients who previously experienced severe immunodeficiency retain low pathogen-specific T-cell responses despite a virological response to antiretroviral therapy (ART). To identify correlates with dysfunction in accessory cell populations, HIV patients were stratified into groups maintaining high or low CD4(+) T-cell IFN-gamma responses to cytomegalovirus (CMV) over 4-8 years on ART. Myeloid dendritic cells (mDC), plasmacytoid (p) DC, M-DC8(+) cells and monocytes were enumerated and mRNA of cytokines and activation molecules were quantitated in purified subpopulations. Proportions of pDC were lower (p=0.043) and mDC were higher (p=0.043) in low responders. TRAIL receptor 2 (DR5) mRNA levels in pDC (p=0.0008) and mDC (p=0.0062) were lower in high responders compared to controls. Levels of IL-15 mRNA were higher in mDC from high responders (p=0.015) and levels of IL-10 mRNA were higher in M-DC8(+) cells from low responders (p=0.036). Hence CMV-specific CD4(+) T-cell IFN-gamma responses may be affected by numbers and function of circulating DC. PMID- 18511345 TI - Monosomy 1p36 uncovers a role for OX40 in survival of activated CD4+ T cells. AB - Monosomy 1p36 is a subtelomeric deletion syndrome associated with congenital anomalies presumably due to haploinsufficiency of multiple genes. Although immunodeficiency has not been reported, genes encoding costimulatory molecules of the TNF receptor superfamily (TNFRSF) are within 1p36 and may be affected. In one patient with monosomy 1p36, comparative genome hybridization and fluorescence in- situ hybridization confirmed that TNFRSF member OX40 was included within the subtelomeric deletion. T cells from this patient had decreased OX40 expression after stimulation. Specific, ex vivo T cell activation through OX40 revealed enhanced proliferation, and reduced viability of patient CD4+ T cells, providing evidence for the association of monosomy 1p36 with reduced OX40 expression, and decreased OX40-induced T cell survival. These results support a role for OX40 in human immunity, and calls attention to the potential for haploinsufficiency deletions of TNFRSF costimulatory molecules in monosomy 1p36. PMID- 18511346 TI - Ag-specific type 1 CD8 effector cells enhance methotrexate-mediated antitumor responses by modulating differentiated T cell localization, activation and chemokine production in established breast cancer. AB - The chemotherapeutic agent methotrexate is widely used in the treatment of breast cancer. Although its mechanism-of-action has been defined, less is known about its interaction with T cell-mediated antitumor responses. Type 1 CD8 T cell mediated immune responses (Tc1) are cytolytic, produce IFN-gamma and are associated with effective antitumor responses. Using a murine transgenic TCR tumor model, we show that single-dose treatment with methotrexate enhanced CD8 mediated type 1 antitumor responses when administered 3 days prior to Tc1 effector cell transfer. Co-treatment with methotrexate not only enhanced donor Tc1 cell accumulation and persistence at sites of primary tumor growth, but also promoted elevated levels of activated donor TIL cells. This markedly enhanced the appearance of endogenous differentiated (CD44(High)) CD8 tumor-infiltrating cells when compared to that of corresponding groups receiving either MTX or Tc1 cell transfer alone. Such cells were acutely activated as defined by co-expression of surface markers associated with TCR engagement (CD69) and T cell activation (CD25) at both early (days 1-8) and late (days 12-20) stages following treatment. Conversely, such animals showed an early decrease in CD4(+)/CD44(High)/CD25(+)/CD69(+) T cells that correlated with delays in tumor growth in vivo. Moreover, cellular response kinetics appeared to further correlate with the up-regulation of endogenous T cells producing the chemokine IP 10 in vivo. This suggested that Tc1 cell transfer, in combination with chemotherapy, can enhance antitumor responses by modulating immunoregulatory T cells involved in homeostasis and immune tolerance within the tumor environment. These studies offer insight into mechanisms that enhance T cell-based immunotherapy in cancer. PMID- 18511348 TI - The other side of the coin: Beneficiary effect of omega-3 fatty acids in sudden unexpected death in epilepsy. AB - The epilepsies are the most common serious neurological condition. People with epilepsy have a two- to threefold increased risk of dying prematurely than those without epilepsy, and the most common epilepsy-related category of death is sudden unexpected death in epilepsy (SUDEP). The exact pathophysiological causes of SUDEP remain unknown, but it is very probable that cardiac arrhythmia during and between seizures plays a potential role. Although the pharmacological treatments available for the epilepsies have expanded, antiepileptic drugs are still limited in clinical efficacy. In this regard, several factors such as genetic, environmental, and social can contribute to the inefficacy of therapeutic outcome in patients with epilepsy. Among these factors, nutritional aspects, that is, omega-3 fatty acid deficiency, have an interesting role in this scenario. Animal and clinical studies have demonstrated that omega-3 fatty acids may be useful in the prevention and treatment of epilepsy. Moreover, as omega-3 fatty acids per se have been shown to reduce cardiac arrhythmias and sudden cardiac deaths, it has been proposed that omega-3 fatty acid supplementation in patients with refractory seizures may reduce seizures and seizure-associated cardiac arrhythmias and, hence, SUDEP. Given their relative safety and general health benefits, our update article summarizes the knowledge of the role of dietary omega-3 fatty acids in epilepsy. PMID- 18511349 TI - Situation-Based Access Control: privacy management via modeling of patient data access scenarios. AB - Access control is a central problem in privacy management. A common practice in controlling access to sensitive data, such as electronic health records (EHRs), is Role-Based Access Control (RBAC). RBAC is limited as it does not account for the circumstances under which access to sensitive data is requested. Following a qualitative study that elicited access scenarios, we used Object-Process Methodology to structure the scenarios and conceive a Situation-Based Access Control (SitBAC) model. SitBAC is a conceptual model, which defines scenarios where patient's data access is permitted or denied. The main concept underlying this model is the Situation Schema, which is a pattern consisting of the entities Data-Requestor, Patient, EHR, Access Task, Legal-Authorization, and Response, along with their properties and relations. The various data access scenarios are expressed via Situation Instances. While we focus on the medical domain, the model is generic and can be adapted to other domains. PMID- 18511350 TI - Evaluation of operational chronic infection endpoints for HCV vaccine trials. AB - Hepatitis C virus (HCV) is a leading cause of chronic liver disease. The natural history of HCV infection is heterogeneous, and a person infected with HCV can clear the virus or progress to a chronic infection. The chronic infection can remain asymptomatic for decades before the development of liver cirrhosis and/or carcinoma. Currently, there are no assays that can differentiate a transient infection (an acute infection that would clear) from a chronic infection, and serial HCV RNA testing is used to operationally define chronic hepatitis C (e.g. detectable HCV over 6 months). Therefore, HCV vaccine trial planning can benefit from the assessment of the endpoint candidates that are aimed at the chronic infection. Operationally defined endpoints based on the virological tests at study visits have been previously studied in the context of human papillomavirus (HPV) vaccine trials. However, HCV natural history is different from HPV, requiring separate considerations. In this work, several definitions of chronic infection that are based on the periodically observed HCV RNA statuses are evaluated, using a multi-state, time-homogeneous Markov model for transient and chronic infections under various infection settings. Our results show some inflation in the type I error in the log-rank test on the vaccine efficacy against chronic infections in the presence of vaccine efficacy related to transient infections. A type I error up to almost four times the planned rate of 5% is observed in one setting. Overall, simple operational endpoints yield higher power than more complex endpoints, but the simplest endpoint is most affected by the type I error inflation and misclassification error due to the assay imperfection. PMID- 18511351 TI - The role of the paratrigeminal nucleus in the pressor response to sciatic nerve stimulation in the rat. AB - The paratrigeminal nucleus (Pa5), an input site for spinal, trigeminal, vagus and glossopharyngeal afferents, is a recognized site for orofacial nociceptive sensory processing. It has efferent connections to brain structures associated with nociception and cardiorespiratory functions. This study aimed at determining the function of the Pa5 on the cardiovascular component of the somatosensory reflex (SSR) to sciatic nerve stimulation (SNS) in paralyzed and artificially ventilated rats following Pa5 chemical lesions (ibotenic acid), synaptic transmission blockade (CoCl(2)), local anaesthetics (lidocaine) or desensitization of primary afferent fibers (capsaicin). The pressor response to sciatic nerve stimulation at 0.6 mA and 20 Hz (14+/-1 mm Hg) was strongly attenuated by contra- (-80%) or bilateral (-50%) paratrigeminal nucleus lesions. Ipsilateral Pa5 lesions only attenuated the response to 0.1 mA, 20 Hz SNS (-55%). Cobalt chloride or lidocaine injected in the contralateral paratrigeminal nucleus also attenuated the SSR. In capsaicin-treated animals, the pressor responses to 0.1 mA were abolished, whereas the responses to SNS at 0.6 mA were increased from 65 to 100% depending on the stimulus frequency. The paratrigeminal nucleus receives both, excitatory and inhibitory components; the later apparently involving capsaicin-sensitive fiber inputs mostly to the ipsilateral site whereas the capsaicin insensitive excitatory components that respond to high or low frequency stimulation, respectively, target the contralateral and ipsilateral sites. Thus, the paratrigeminal nucleus mediates excitatory and inhibitory components of the somatosensory reflex, representing a primary synapse site in the brain for nociceptive inputs from the sciatic innervation field. PMID- 18511352 TI - Peculiarities of RBCs resistance to acid hemolysis in hibernating mammals. AB - The binding of acids and alkalis, formed in tissues by metabolism, along with oxygen and CO(2) transport are recognise as the principal functions of red blood cells (RBCs). Decreases in internal environment pH may result in activation of potential endogenous cytotoxic metabolites, OH and oxidant formation and, as a consequence, result in oxidative damage of cell membrane leading to hemolysis. The characteristics of acid hemolysis in hibernating mammals have been determined in this study. Parameters of HCl-hemolysis, such as the average time for RBC hemolysis and the population distribution of the response, have been investigated. Measurements were performed within 40-5 degrees C temperature range. The resistance of hibernator RBCs to increased acidity, determined according to the acid hemolysis parameters, was found to reflect whether the animal was in the summer active period or in hibernation, with differences also apparent at different points with the hibernation season itself. It was demonstrated that hemolysis parameters of naturally cold-tolerant organisms are altered by decreases in temperature. We discuss a role of cytoskeletal-membrane interactions as a fast-acting switch of the structural and functional state of hibernator RBCs as an adaptation mechanism to acidosis arising from hypothermia and hypermetabolism. PMID- 18511353 TI - Dielectrophoretic sorting on a microfabricated flow cytometer: label free separation of Babesia bovis infected erythrocytes. AB - Dielectrophoresis is a method that has demonstrated great potential in cell discrimination and isolation. In this study, the dielectrophoretic sorting of normal and Babesia bovis infected erythrocytes was performed using a microfabricated flow cytometer. Separation was possible through exploitation of the dielectric differences between normal and infected erythrocytes, essentially due to the higher ionic membrane permeability of B. bovis infected cells. Sorting experiments were performed inside a microchip made from Pt microelectrodes and SU 8 channels patterned on a glass substrate. Optimum cell separation was achieved at 4 MHz using an in vitro culture of B. bovis suspended in 63 mS/m phosphate buffer and applying a sinusoidal voltage of 15 V peak-to-peak. Normal erythrocytes experienced stronger positive dielectrophoresis (pDEP) than B. bovis infected cells, moving them closer to the microelectrodes. Under these conditions it was possible to enrich the fraction of infected cells from 7 to 50% without the need of extensive sample preparation or labelling. Throughout the experiments very few microliters of sample were used, suggesting that this system may be considered suitable for integration in a low-cost automated device to be used in the in situ diagnostic of babesiosis. PMID- 18511354 TI - Permeability of rat and rabbit erythrocyte membranes for a series of amides. AB - Permeability coefficients of rat and rabbit erythrocyte membranes for a series of amides, as well as for erythrocytes treated with p-chloromercuribenzenesulfonic acid monosodium salt (pCMBS) have been determined at 25 and 37 degrees C. Directly proportional dependence of the pCMBS treated erythrocyte permeability for investigated substances and their partition coefficients between the hydrophobic phase and water as well as the values of activation energy of this process indicate that penetration of small hydrophilic molecules is realized by passive diffusion through the lipid bilayer. The results obtained indicate that penetration of small hydrophilic molecules of formamide through lipids is determined by the existence of a free space between hydrocarbon chains that arises from kink formation. The differences in permeability between rat and rabbit erythrocyte membranes could arise in particular as a result of the differences in lipid composition. PMID- 18511355 TI - Efficacy of high dose phylloquinone in correcting vitamin K deficiency in cystic fibrosis. AB - BACKGROUND: Subclinical deficiencies of vitamin K are universally present in unsupplemented cystic fibrosis (CF) patients. The dose required to prevent deficiencies cannot be estimated from the existing literature. The aim of this study is determine if a supplemental dose of 1 mg/day or 5 mg/day vitamin K1 per day would normalize vitamin K status in a population of children with cystic fibrosis. METHODS: Fourteen pancreatic insufficient CF children, between the ages of 8 to 18 years old, were randomized to receive either 1 mg/day or 5 mg/day vitamin K1 per day, for one month. Fasting blood tests were done at baseline and after one month of the intervention. The degree of undercarboxylation of osteocalcin (%Glu-OC), and serum vitamin K1, were evaluated by descriptive statistics and nonparametric Wilcoxon matched-pair test and Mann-Whitney U test. RESULTS: Of the 50% of subjects who were below the optimal serum vitamin K1 at baseline, all rose into the normal range with supplementation. Supplementation also significantly reduced the overall %Glu-OC from a median of 46.8 to 29.1% (p<0.0003). CONCLUSION: Our results suggest that both 1 mg and 5 mg of vitamin K1, given over a one-month period in pancreatic insufficient pediatric cystic fibrosis patients improve vitamin K status. PMID- 18511357 TI - Quantitative profiling of bile acids in biofluids and tissues based on accurate mass high resolution LC-FT-MS: compound class targeting in a metabolomics workflow. AB - We report a sensitive, generic method for quantitative profiling of bile acids and other endogenous metabolites in small quantities of various biological fluids and tissues. The method is based on a straightforward sample preparation, separation by reversed-phase high performance liquid-chromatography mass spectrometry (HPLC-MS) and electrospray ionisation in the negative ionisation mode (ESI-). Detection is performed in full scan using the linear ion trap Fourier transform mass spectrometer (LTQ-FTMS) generating data for many (endogenous) metabolites, not only bile acids. A validation of the method in urine, plasma and liver was performed for 17 bile acids including their taurine, sulfate and glycine conjugates. The method is linear in the 0.01-1 microM range. The accuracy in human plasma ranges from 74 to 113%, in human urine 77 to 104% and in mouse liver 79 to 140%. The precision ranges from 2 to 20% for pooled samples even in studies with large number of samples (n>250). The method was successfully applied to a multi-compartmental APOE*3-Leiden mouse study, the main goal of which was to analyze the effect of increasing dietary cholesterol concentrations on hepatic cholesterol homeostasis and bile acid synthesis. Serum and liver samples from different treatment groups were profiled with the new method. Statistically significant differences between the diet groups were observed regarding total as well as individual bile acid concentrations. PMID- 18511358 TI - Polypropylene mesh covered colonic anastomosis. Results of a new anastomosis technique. AB - BACKGROUND: The morbidity and mortality rates of anastomosis leakage of the gastrointestinal system, are high. In this study we covered the colonic anastomosis with polypropylene mesh on the safety of the anastomosis was investigated. METHODS: Twenty female albino rabbits were divided into two groups. First of all, a segmental colon resection was performed in both the groups and a single layer of anastomosis was made. In addition, a polypropylene mesh as long as the circumference of the anastomosis in the study group. All the rabbits were sacrificed on the 10th postoperative day and the explosion pressure of the anastomosis, histopathological investigation of the anastomotic contour, and peritoneal adhesion were compared. RESULTS: The anastomoses of all the subjects in the control group had exploded and the average explosion pressure was 149 +/- 16 mmHg. However, in the study group, the anastomoses did not explode in nine (90%) of the subjects, whereas it exploded in only one (10%) with a pressure of 260 mmHg. The average explosion pressure in the study group was 315 +/- 30 mmHg (p < 0.0001). No significant difference was established between the groups according to the histopathological classification of the anastomotic contour performed according to the Ehrlich-Hunt model (p > 0.05). Peritoneal adhesions of the groups is not statistically different (p > 0.05). CONCLUSION: During the short follow-up period, this new technique significantly increased the safety of the anastomosis, moreover it did not cause any increase in peritoneal adhesions. This success has most probably occurred as a result of the external mechanical support to the anastomosis. PMID- 18511356 TI - Biomechanics of liquid-epithelium interactions in pulmonary airways. AB - The delicate structure of the lung epithelium makes it susceptible to surface tension induced injury. For example, the cyclic reopening of collapsed and/or fluid-filled airways during the ventilation of injured lungs generates hydrodynamic forces that further damage the epithelium and exacerbate lung injury. The interactions responsible for epithelial injury during airway reopening are fundamentally multiscale, since air-liquid interfacial dynamics affect global lung mechanics, while surface tension forces operate at the molecular and cellular scales. This article will review the current state-of knowledge regarding the effect of surface tension forces on (a) the mechanics of airway reopening and (b) epithelial cell injury. Due to the complex nature of the liquid-epithelium system, a combination of computational and experimental techniques are being used to elucidate the mechanisms of surface-tension induced lung injury. Continued research is leading to an integrated understanding of the biomechanical and biological interactions responsible for cellular injury during airway reopening. This information may lead to novel therapies that minimize ventilation induced lung injury. PMID- 18511359 TI - Echocardiographic measurements of cardiac dimensions in normal Standardbred racehorses. AB - OBJECTIVES: The aim of this study was to obtain echocardiographic measurements and establish reference ranges for 14 parameters in Standardbred racehorses in training. BACKGROUND: Several studies have been published about cardiac measurements in Thoroughbreds, Standardbreds, National Hunt horses, Warmbloods and ponies; however, not all parameters have been published for the Standardbred trotter in training. ANIMALS, MATERIALS AND METHODS: Thirty normal Standardbred racehorses in training were assessed by two-dimensional echocardiography (2-D) and M-mode echocardiography using standardized imaging planes. Mean values, standard deviations, 95% confidence interval for the means and 95% confidence interval for the cardiac parameters measured in the population were calculated. Furthermore, a general linear model was constructed using sex, age and body weight (bwt) of the horses as independent variables and the echocardiographic measurements as dependent variables. Multivariate linear regression analysis was performed with the level of significance at p<0.05 for all the null hypotheses. RESULTS: Reference ranges were established for 14 echocardiographic parameters in Standardbred racehorses. Weak linear relationships between echocardiographic measurements and body weight were observed for LVIDd, LVIDs, LVFWs, and AOD. Linear regressions on these parameters were used to calculate the 95% confidence intervals for the predicted values. CONCLUSIONS: The data collected in this study provide reference values for the evaluation of Standardbred racehorses in training. Body weight has a negligible affect on most echocardiographic parameters in this homogeneous population, but did mildly influence the results of left ventricular and aortic measurements. PMID- 18511360 TI - [Frequency of cancer at older ages]. AB - The dependency between the risk of death and age is analysed, and the contribution of cancer to the overall risk of death is evaluated as a function of age. The frequency of the different cancer sites is described in different age groups. Lastly cancer mortality trends are presented by age. The risk of death from cancer increases markedly with age, but the risk of a death from a cardiovascular disease increases even more rapidly, consequently the importance of cancer as a cause of death decreases with age. In the male population, lung and head and neck cancers are the most frequent cancers before age 65, whereas prostate and colorectal cancers are more frequent at older ages. In the female population, breast and colorectal cancers are the most frequent cancers except for mortality before age 65 where lung cancer is the second killer after breast cancer. The risk of cancer death decreases in recent years for all age groups. PMID- 18511361 TI - [Aging and geriatric oncology]. AB - Chronologic age is not the exclusive factor for therapeutic decision, specially because life expectancy in very old age remains significant. Evaluation of health, autonomy and diseases are more important. Health status in very old age results from genetic factors, life-course and aging. The later may be understood as an adaptative process in response to life-course. These factors lead to frailty, whose evaluation is essential to high quality care in elderly patients. This evaluation must be multidimensional and lead to a care-plan. Even no randomized controlled trial had demonstrated benefits of geriatric intervention in cancer elderly patients, there is a high probability that improvement in mortality, morbidity, autonomy and quality of care may be obtained by strong collaboration of oncologists and geriatricians. PMID- 18511362 TI - [Geriatric assessment in the oncology setting]. AB - The optimal management of elderly patients with cancer includes, besides the tumour assessment, a geriatric evaluation procedure. To date, comprehensive geriatric assessment (CGA) remains the gold standard of evaluation in older patients. To be effective, the process must be performed by a team with special expertise in caring for elderly people, in a specific geriatric unit. Alternative procedures have been developed and studied in the oncology setting, multidimensional geriatric assessment and screening tools. This approach has been developed to help oncologists differentiate the healthy elderly from other older patients whose problems would possibly interfere with cancer treatment and who require a pre-treatment CGA. Studies have shown that healthy older patients derive no advantage from CGA, but benefit from cancer treatment as much as younger patients. PMID- 18511363 TI - [Pharmacology of anticancer drugs in the elderly: tools for dose-adjustment]. AB - Life expectancy increasing and cancer incidence rising with age, geriatric and cancer care will become a significant medical, public health, challenge. It is possible that the lack of efficacy of cancer therapies in the elderly may simply be due to the fact that physicians reduce anticancer drug doses empirically, in order to avoid "putative" toxicities that might arise as a result of alterations of physiological functions or as a result of co-morbidities generally present within this population. However, many authors have demonstrated that some patients over 70 years old could tolerate and obtain same benefit from therapies as younger adults, when some who are frail need less aggressive therapies. It is imperative that decisions regarding the management of cancer in older persons should be based upon the individual needs and fitness of the patient, and not based on chronological age. The main difficulty is the lack of scientific references on optimal treatment-dosing in the elderly, and we cannot extrapolate, as it stands, the information from younger patient. Indeed, aging is associated with multidimensional changes, including alteration of physiological status, comorbidities and polymedications. These changes may lead to pharmacokinetic (PK) modifications, namely in absorption, distribution, metabolism and elimination of drugs as well as pharmacodynamic (PD) modifications. Prospective studies need to be implemented in the elderly in order to study in depth the PK and PD properties of the drugs used for these patients, and establish PK-PD relationships in this specific population. Such studies have been successfully conducted in the elderly, some of them leading to dose recommendations. This paper detail the different sources of PK-PD variability in the elderly, some practical considerations regarding the design of studies using the population approach, as well as some examples of studies performed in the elderly. We conclude with some recommendations in this population at risk. PMID- 18511364 TI - [Chemotherapy in the elderly: how and for whom?]. AB - Management of cancer in the older-aged patient is an increasingly common problem in our occidental societies. Cancer is a disease primarily of older persons: over 60% of all cases of cancer are diagnosed after age 65 - an age group that constitutes less than 20% of the western population and the risk of persons over 65 years of age developing cancer is at least 10 times that of those under 65. Cancer in older persons may be considered a different disease from cancer in the younger in that way that biology of the host could influence the growth of cancer, that the management of the disease deserved an individualized approach. Indeed, the normal process of aging is associated with a progressive age-related reduction in function of many organs, including losses such as renal, pulmonary, cardiac, immune, hepatic, haematological, muscles, osseous, sight, hearing and brain functions. The consequences of these changes with age, added to comorbid diseases, have major implications on toxicities of anti-cancer therapies, surgery, radiotherapy as well as chemotherapy. However chronologic age should not be used as a guide to cancer therapy. Performance status and physiologic performance of the older patient are of prime importance to decide and conduct chemotherapy. PMID- 18511365 TI - [Anticancer chemotherapy in the elderly: a review of the literature]. AB - Few elderly cancer patients are included in clinical trials, resulting insuffisant data of the effectiveness and tolerance of anticancer drugs in this patient population. The aim of this study was to analyse the studies concerning the effectiveness and tolerance of chemotherapy prescribed for elderly patients treated for colorectal, breast and lung cancer. The data of this population showed that the older patients are less likely to receive chemotherapy than the younger. The age is considered as significant important factor for the decision of chemotherapy of this population. However elderly patients seem to have the same benefit as compared with younger patients. Rather than the criteria of age, comorbidities should be considered. It is necessary to develop specific geriatric assessment and development of clinical trials specifically including elderly patients remains a necessity. PMID- 18511366 TI - [Going through chemotherapy: an age-revealing process]. AB - Elderly persons want to know about their diagnosis and therapeutic alternatives. Feeling in good health before as well as having gone through hard times enhance acceptability of chemotherapy. Possible issues are perceived in a limited but often overestimated lap of life. At this time, actual age is not present in the patient's mind. But as a result of the treatment, patients feel signs of ageing. The repercussions of the chemotherapy, mainly the tiredness, are withdrawal from social life and retreat into the private sphere. Cancer and chemotherapy give rise to a feeling of loneliness and vulnerability never known until this stage. Weakened by the experience, patients feel ageing. Transport difficulties and isolation are two significant consequences. Although patients claim to have taken the decision to be treated on their own, the investment of the closer relatives will prove to be essential during the chemotherapy. One cannot think about chemotherapy for elderly people without integrating psychosocial and economic dimensions from the outset. The geriatric evaluation must be seen as a dynamic approach likely to be modified by integrating new elements at any stage of the patient's treatment. PMID- 18511367 TI - [Breast cancer in the elderly]. AB - Breast cancer is the most commonly diagnosed cancer and leading cause of cancer mortality in women worldwide. The elderly comprise a large part of the breast cancer population, and there are important specific considerations for this population. Late diagnosis and substandard local and systemic therapies are frequent, which is only partially "compensated" by a more indolent tumour behaviour due to the increasing likelihood according to age of potentially hormone sensitive tumour status. Endocrine treatment remains a key component of systemic treatment in both advanced and early setting. However chemotherapy is a valid option, with interest strengthened by proven efficacy in adjuvant setting for aggressive phenotypes, better management of side effects and attempts to develop predictive index for toxicity. The recently reported laboratory studies on potential mechanisms for resistance to endocrine therapies that involve crosstalk between growth factor signalling pathways and hormonal receptors stimulate also new therapeutic approaches. PMID- 18511368 TI - [Lung cancers in the elderly]. AB - Lung cancer is the leading cause of cancer-related death worldwide. The world population is getting increasingly older. The incidence of lung cancer is therefore growing among elderly patients. However, in spite of the universal recognition of the importance of clinical research to guide therapeutic decisions, elderly lung cancer patients are largely under-represented in cancer treatment trials. Age alone must not be a limiting factor in the choice of treatment. Indications for surgery should not vary with age. Fit elderly patients benefit from concurrent chemoradiotherapy. For metastatic patients, single-agent monotherapy with a third-generation agent (vinorelbine, gemcitabine or docetaxel) is the recommended option. Platinum-based chemotherapy may represent a valid option for fit elderly patients with adequate function. The role of the new biologic target-based agents is to be found. There is an absolute need of clinical trials specifically dedicated to elderly patients with lung cancer. PMID- 18511369 TI - [Prostate carcinoma in the elderly]. AB - Prostate carcinoma is the most frequent cancer in French men. Prostate carcinoma in elderly is the topic of this review. The review included chapters on age as the main factor for prostate carcinoma, the modification of disease presentation or PSA level depending on age, and then the challenge of age in therapeutic decisions. Finally, we evaluate the place of elderly patients in prospective trials and in adjustable therapies. PMID- 18511370 TI - [Colorectal cancer in the elderly]. AB - Colorectal cancer occurs after the age of 75 in 50% of the case, but elderly patients are under-treated and far from an optimal management of their disease. Principles of colorectal cancer management after the age of 75 are theoretically the same than for younger patients, but physiological age, comorbidities and patient's wishes should be taking into account. Treatment should be individually based and discussed during multidisciplinary meeting with oncologists and geriatrics after a complete geriatric evaluation. Surgery remains the only curative treatment and should be discussed when feasible. PMID- 18511371 TI - [Chronic lymphoproliferative disease in the elderly]. AB - Chronic lymphoproliferative disorders represent a group of malignant lymphoid diseases of whom chronic lymphoid leukaemia (CLL) is the more frequent. They are incurable diseases of indolent evolution. Diagnosis is usually made on blood puncture when circulating malignant lymphocytes are present. Explorations are limited to CT-scan, and, for CLL, karyotype. Bone marrow biopsy must be avoided, as possible. Treatment indications will be limited to really evolved cases. The aim of treatment must be the quality of life. Adapted treatment are available for the majority of patients, with limited toxicity and prolonged efficacy. PMID- 18511372 TI - [Non-Hodgkin lymphoma in the elderly]. AB - Non-Hodgkin's lymphomas form a heterogeneous group of tumors whose incidence is rising in elderly subjects. While most cases are accessible to treatment with chemotherapy, elderly patients tolerate treatments less well than younger ones. Nevertheless, treatment efficacy does not seem different. Two approaches can thus be proposed: the first is to give the patient a conventional treatment with a higher risk of toxicity, the second one would be to propose a "lighter", dose adapted treatment. Two examples are examined more closely: diffuse large B-cell lymphomas and follicular lymphomas. PMID- 18511373 TI - [Treatment of myeloma in the elderly]. AB - The treatment of elderly patients with multiple myeloma was based for the last forty years on the combination of alkeran plus prednisone. The survival was short. Three new molecules, thalidomide, bortezomib and revlimid are completely changing the way we treat myeloma as well as the prognosis. These new molecules are tested either alone or in combination. The addition of thalidomide to alkeran plus prednisone has increased overall survival of two years. For the first time, major therapeutic improvements are emerging giving rise to better prognosis. PMID- 18511374 TI - [Chemotherapy-induced cardiotoxicity in the elderly]. AB - Chemotherapy-induced cardiotoxicity can concern major effects, inducing severe impairments of vital functions: systolic or diastolic function, hypertension, rhythmic or conducting pathology, Elsewhere, cardiac and vascular aging induces alterations, which concern roughly the same points and are enhanced by vascular risk factors. We wish to analyse the correlation and increase of the consequences of the first one toward the second one and the safe attitude we must have for those patients (prevention and early treatment). Echocardiography seems to have more and more a major status to assess cardiac function, chiefly in systolic and diastolic manor. We insist on the interest of stress echo for assessment of impaired contractility and for evaluation of vascular risk in ischemic disease. We suggest a vulnerability score to predict the risk of complication due to chemotherapy. PMID- 18511375 TI - [Chemotherapy and renal toxicity]. AB - Antineoplastic drugs used in the treatment of cancers present with variable renal tolerance profiles. Among drugs with a potential for renal toxicity, platinum salts, methotrexate, and gemcitabine are well-known. The mechanisms of their renal toxicity and the means of its prevention are presented in this article. Anti-angiogenic drugs, recently marketed or still under clinical development, may also interact with the kidneys. In general, optimising the renal tolerance of anticancer drugs requires an appropriate evaluation of patients'renal function, before and during treatment, at each course. Serum creatinine alone is not a reliable index of renal function. Its evaluation must be performed with the use of Cockcroft-Gault or aMDRD formulae. In patients with abnormal renal function, dosage adjustment is often required to improve the renal tolerance, and also to limit the risk of extra-renal toxicities (such as haematological toxicities) induced by a drug overdosage, in those patients with reduced drug-elimination. PMID- 18511376 TI - [Development of oncogeriatry: national and international strategies]. AB - Incidence of cancer increases with the recent increase of the life expectancy. Aging is characterized by health status heterogeneity. It is therefore difficult to exactly predict tolerance to treatment and to make decision. One of the major question is to classify the problems on the base of their impact on health status and outcome. It is only possible if oncologists and geriatricians work together. The objectives at the national and international levels are the same: to develop formation (teaching programs), information (for professionals and the public), clinical research. It is more difficult to develop case management, because it depends on health organisation status. A strong global program has been developed in France with the emergence of Pilot Units for the coordination of Oncogeriatrics. It begins to produce results. PMID- 18511377 TI - [Radiation therapy in the elderly]. AB - Cancer is a disease that predominantly occurs in older patients who represent a quarter of the population in western countries. Numerous types of cancer are observed in elderly people. Radiotherapy is one of the most powerful treatment against cancer. The purpose of this review is to highlight sights of radiation oncology specifically related to aging. Particular emphasis is placed on logistic and technical aspects of radiation. Special techniques are also reviewed that have particular relevance to the treatment of the elderly, as modulated intensity radiotherapy, gating, cyberknife use and partial breast irradiation. Most of published studies have demonstrated feasibility of radiotherapy in curative or palliative intent whatever cancer types are considered. Complete geriatric assessment and a multidisciplinary approach are the key points. PMID- 18511378 TI - [State of nutrition and cancer in the elderly]. AB - Malnutrition is a common problem in cancer patients, associated with an increased risk of complications and a reduced response to treatment resulting to a poorer outcome. Both age and oncological disease can affect the state of nutrition in multiple ways. In elderly, malnutrition can be affected either directly by the tumour process (local effect, host response to the tumour ...), by the adverse effects of the treatment, and by the impact of several psychosociological and physiological factors due to aging. Prophylactic interventions in all these stages should therefore be done, addressing the specific needs of this patients group, and suggesting the systematic use of an out-patient screening tool (MNA). Assessing and modifying nutrition must be an integral part of the treatment. PMID- 18511379 TI - [Oncogeriatry and psychology: an other glance at the elderly]. AB - Medicine, as well as the whole society, should cope with a society getting older and older. Yet, to what extend are the peculiarities of elderly assessed and taken into account? And what about relationship and communication? Are elderly offered similar therapeutic projects and are they equally informed about them? Do they have, themselves, the same needs? Oncologists and the nursing staff encounter several peculiar difficulties while coping with elderly. These difficulties are not only due to a complicated medical situation of a multi treated patient with along history of many other diseases, but also to strongly anchored received ideas that "it's easier to die when one is old" and that "it's better to preserve the patient by hiding his disease to him, and by lying to him about it, because it is worthless to tell him the truth". Yet, these ideas are not of any help. Wrong ideas, lack of information, means and trained manpower and sometimes of interest affect often negatively the patient's treatment outcome, and highlight, more than in any other patients, psychological, relational but also ethical thorny stakes. By focusing on the elderly peculiarities, while considering them as patients on their own, it would be possible to highlight these stakes and to better assess these touch by situations. This would help enhancing a better use of neuropsychology and psycho oncology and by that better tailor the therapeutic and nursing decisions as well as communication fashions in oncogeriatry. PMID- 18511380 TI - [The role of computed tomography perfusion in the diagnosis of brain ischaemic stroke]. AB - In recent years, the broad introduction of fast multidetector computed tomography (CT) systems and the availability of commercial software for perfusion analysis have made cerebral perfusion imaging with CT a practical technique for the clinical environment. AIM AND METHODS: This article reviews the use of CT for imaging cerebral perfusion, highlighting its advantages, disadvantages and limitations, and draws comparisons between perfusion CT and magnetic resonance imaging. The authors performed 96 perfusion CT examinations in the last one and a half years. Future technical developments in multi-slice CT systems may diminish the current limitations of limited spatial coverage and radiation burden. Yet CT is often not perceived as a technique for imaging cerebral perfusion. CONCLUSIONS: The technique is widely available at low cost, accurate and easy to perform. Perfusion CT is particularly applicable to those clinical circumstances where patients already undergo CT for other reasons, including stroke. PMID- 18511381 TI - [Implantation of subcutaneous defibrillator in a patient treated for recidive endocarditis, previously operated for tetralogy of Fallot]. AB - Implantable cardioverter-defibrillator implantation is an important adjunct in the management of these patients, although cardiac device endocarditis is an infrequent, but potentially lethal infectious complication. Cure of cardiac device endocarditis is achievable in the large majority of patients treated with an aggressive approach of combined antimicrobial treatment and complete cardiac device removal. Re-implantation of the device requires additional caution, but still associated with increased risk for recurrences. AIM: The aim of this case presentation is to demonstrate the feasibility of permanent subcutaneous defibrillation system in a patient with recurrent endocarditis. METHOD: In our case a 37-year-old patient--operated several times due to Fallot tetralogy, with permanent pacemaker and later implantable cardioverter defibrillator, requiring several replacements--was treated with recurrent device endocarditis. After device explantation and long-term standard antibiotic treatment, in order to decrease further risk for the development of recurrent endocarditis, we implanted a defibrillation system with an individually designed subcutaneous lead configuration. CONCLUSION: This case report suggests that individually tailored defibrillator lead configuration is a feasible option for patients until a fully automatic effective subcutaneous system will be available. PMID- 18511382 TI - [Role of the Orvosi Hetilap in the development of Hungarian gastroenterology. Part 3: 1948-1989]. AB - The editions of Orvosi Hetilap published between 1905 and 1944 exceeded the amount of publications of the previous period (1857-1904). Most of the knowledge was transferred from German literature. AIM: Evaluation of papers dealing with diseases of the gastrointestinal tract published in Orvosi Hetilap between 1948 and 1989. METHOD: The author manually reviewed the journal volumes published between 1948 and 1989. The original articles, journal and book reviews were identified and classified according to their subject and origin. The publication rates of the editorial periods (1857-1904, 1905-1944, 1948-1989, respectively) were statistically compared. RESULTS: Between 1948 and 1989, a total of 2175 original articles were published about the diseases of the gastrointestinal tract, constituting 11.1% of the total publications. There was a continuous increase in the publication rate in the periods studied. Liver and biliary tract diseases were most often studied (24.1%), especially viral hepatitis (18.0%). Endoscopy was studied in 124 articles (4.7%), while radiology achieved a figure of 101 (3.9%). 3965 articles were reviewed from 268 journals, constituting a significant increase as compared to the previous period. 62.2% of the reviews were published in 15 core journals. 52.2% of the journals were English, 39.4% were German, and only 2.9% French. The formerly major German sources were replaced by English literature during the 1960s. The number of book reviews increased from 95 to 211. Endoscopy witnessed the emergence of fiberoptic instruments and, later, video technique. The reception of inflammatory bowel diseases began in 1935 and expanded gradually in the epidemiologic, pathogenetic and therapeutic studies of these conditions. CONCLUSION: The rate of gastroenterologic publications in Orvosi Hetilap increased considerably between 1948 and 1989, as compared to the prior periods. English journals became the main source of information in the 1960s. PMID- 18511383 TI - [Vena portae obstruction as a cause of portal hypertension in a child with Turner syndrome]. AB - Cardiovascular and renal malformations are well-known in Turner syndrome. However, gastrointestinal bleeding is less frequent. The possible etiologies of gastrointestinal bleeding in Turner syndrome are intestinal teleangiectasia, inflammatory bowel disease and portal hypertension. The authors report a 3-year old girl with Turner syndrome who presented with severe gastrointestinal bleeding requiring transfusion. The radiological examination indicated prehepatic portal hypertension as a reason for haematochezia. The liver biopsy demonstrated the anomaly of intrahepatic arteries and veins. In this report we describe a case of congenital portal vein obstruction and we reviewed liver abnormalities associated with Turner syndrome and causes of obstruction of vena portae in childhood. PMID- 18511384 TI - [Morphological and functional characteristics of the left ventricle in athletes of various ages, and performing at various levels]. AB - One of the most important effects of regular physical training is the adaptation of the cardiovascular system. The basic importance of an "athlete's heart" is manifested in two fields, one is public health, the other competitive sport. AIM: 1. Can the higher E/A quotient of physically active persons be explained by the training bradycardia, or can it be supposed as an independent effect of regular physical exercise? 2. Which training-induced heart characteristics seem to be different in players of various ball-games? 3. How are cardiac data related to the relative aerobic power as most indicative index of endurance performance? METHODS: Two-dimensionally guided M-mode and Doppler echocardiographic data of different athletic and non-athletic subjects were compared with each other, differences between data of different athletic groups were also analysed. RESULTS: 1. E/A quotient characterises left ventricular (LV) diastolic function, but it highly depends on the heart rate. The higher E/A found in young athletes does not seem to be an independent effect of the regular physical training. 2. Investigating the data of different ball-game-players, characteristic differences were seen. In the myocardial hypertrophy and in the resting heart rate water polo players were the best, volleyball players the worst, but the values of the latter were also better than those of non-athletes. 3. In the pooled group of non athletes and of competitors of several kinds of sports (endurance athletes, ball game-players, power-and-sprint-events athletes) all measured parameters correlated significantly with the relative maximal oxygen consumption. CONCLUSIONS: 1. The higher E/A in elderly subjects suggests that regular physical training can diminish the age-associated impairment of diastolic function. 2. For the high intensity training and for the long competition period a special endurance training program would be useful for volleyball players. 3. The higher was the proportion of endurance activity in the training and competitive program, the stronger were the correlations with the relative aerobic capacity. PMID- 18511385 TI - [Treatment of urosepsis]. PMID- 18511388 TI - Epidemiologists, our patient is society. AB - The common approach utilized in many epidemiologic studies is to measure and compare characteristics and exposures of afflicted and non-afflicted individuals from the same population. The subject of most contemporary chronic disease epidemiology, therefore, is not epidemics per se, nor their causes, but rather epidemics' victims. While these approaches and methods are entirely appropriate for investigating many vitally important health-related questions, it is argued here that epidemics themselves, including those that produce chronic illnesses, should also be the subject of epidemiologic investigations. Epidemics are viewed as phenomena that afflict social entities and should be approached as such. PMID- 18511389 TI - Response: if society is our patient, how shall we take its pulse? AB - A society's health status is not simply the sum of the health of individuals, but an emergent property. While epidemiology has evolved from the study of simple epidemics to the study of the determinants of population health, it is still appropriate to consider the population as the epidemiologist's patient. At the same time, the population is also our study subject-we study populations to identify ways to prevent disease in populations. This article illustrates some of the implications for the prevention of disease that result from treating society as our patient. PMID- 18511390 TI - One foot in, one foot out. AB - As radical health professionals, we have the triple identity of workers, of activists, and of intellectuals that creates the cauldron in which we live contradictory lives. We share the concerns of other workers for salaries, job security, health and safety at work, and work load. But like teachers and unlike most other workers we are not completely alienated from our work and have a stake in the outcomes of our labors that we are not always free to express or act on. As professionals, we share with our colleagues the intellectual curiosity about the origins of new infectious diseases, how racism exhausts the adrenals, or the egg-laying behavior of mosquitoes in polluted environments but may be in conflict with them around the need for universal free health care, or distorted priority given to molecular approaches to disease. We are partisans in institutions that feign neutrality. As activists, we are unwilling to accept the boundaries of "professionalism" and both learn from and offer our understanding to our communities of struggle. These three moments of our lives sometimes conflict but are also mutually enriching. There is no single formula for navigating this turbulent habitat but there is an array of options for avoiding burn-out and enjoying a life that is politically productive, intellectually challenging, and with lovely people. PMID- 18511391 TI - Self-reported health effects among community residents exposed to perfluorooctanoate. AB - Serious health effects due to perfluorooctanoate (PFOA) exposure are suspected. The aim of this study was to evaluate the health status of nearby residents, with prolonged exposure to PFOA in their drinking water. A population of 566 white residents who were plaintiffs or potential plaintiffs in a lawsuit was evaluated by questionnaire for health history and symptoms. Standardized Prevalence Ratios were estimated using National Health and Examination Survey (NHANES) data files for comparison rates. The exposed subjects reported statistically significant greater prevalence of angina, myocardial infarction, and stroke (SPR=8.07, 95% C.I.=6.54-9.95; SPR=1.91, 95% C.I.=1.40-2.62, and SPR=2.17, 95% C.I.=1.47-3.21, respectively), chronic bronchitis, shortness of breath on stairs, asthma (SPR=3.60, 95% C.I.=2.92-4.44; SPR=2.05, 95% C.I.=1.70-2.46; SPR=1.82, 95% C.I.=1.47-2.25, respectively), and other serious health problems. The increased prevalence of adverse health effects may be due to PFOA. Further study is needed. PMID- 18511392 TI - Mad cows and computer models: the U.S. response to BSE. AB - The proportion of slaughtered cattle tested for BSE is much smaller in the U.S. than in Europe and Japan, leaving the U.S. heavily dependent on statistical models to estimate both the current prevalence and the spread of BSE. We examine the models relied on by USDA, finding that the prevalence model provides only a rough estimate, due to limited data availability. Reassuring forecasts from the model of the spread of BSE depend on the arbitrary constraint that worst-case values are assumed by only one of 17 key parameters at a time. In three of the six published scenarios with multiple worst-case parameter values, there is at least a 25% probability that BSE will spread rapidly. In public policy terms, reliance on potentially flawed models can be seen as a gamble that no serious BSE outbreak will occur. Statistical modeling at this level of abstraction, with its myriad, compound uncertainties, is no substitute for precautionary policies to protect public health against the threat of epidemics such as BSE. PMID- 18511393 TI - The promise of engaging in "deep" partnerships. PMID- 18511394 TI - Community-university partnerships: achieving the promise in the face of changing goals, changing funding patterns, and competing priorities. AB - An important challenge that community-university partnerships face is how to maintain themselves in the face of changing goals, priorities, and funding. Partnerships often form as a result of some sort of "spark:" an incident, perhaps, or the identification of a shared need or common concern. Often, external funding is sought to provide the majority of resources for the establishment of a partnership and for the implementation of the partnership's action plan. Whatever external funding is obtained is typically of short duration. The funding will not continue over time. And usually the funding comes with stipulations about allowable partnership approaches; inevitably the priorities of one funder will differ from those of another. These issues of the maintenance of partnership in the face of shifting funding and priorities are ones that confront most community-university partnerships. This article examines these issues through the lens of an environmental justice partnership that has existed for nearly a decade, has undergone many changes in who is involved, and has operated with funding from many different sources, including the National Institute of Environmental Health Sciences, Housing and Urban Development, and the Environmental Protection Agency. In addition, the entities in the partnerships, while they share certain environmental objectives, are sometimes at odds on particular goals when the needs of the agencies differ. Further complicating this issue of continuity is the reality that partners, on occasion, must shift priorities after partnership goals are established. The experiences of this environmental justice partnership shed light on the kinds of struggles community-university partnerships face when they hope to avoid being undermined by the larger concerns of the funders or by the power brokers in their individual organizations. This article examines approaches that community-university partnerships might take to remain resilient in the face of changing goals, priorities and funding. PMID- 18511395 TI - Learning to listen: designing architectural education through university community partnerships. AB - Our world is shaped by a patchwork of perspectives, of stories told to construct or even recreate our realities. Practitioners engaging with the built environment have a responsibility to assist in the proper translation of these stories into something that physically enriches the user's sense of place. However, academia separates itself from the "real world" in order to teach theory, history, and so forth as purely as possible. Critics argue that this separation is problematic when preparing students for a practical field in which they will become heavily entrenched in this world from which they have been sheltered. In response, community-based design initiatives are forming that conjoin students, faculty, community members, and activists to address urgent needs in neighborhoods around the globe. While empowering the communities through the opportunity to change their own surroundings, the researchers benefit from a culturally significant palette with which to search for innovative ways to make the built environment truly relevant to positive transformation at the local level. Nonetheless, this pedagogical method is, in turn, criticized for blurring the line between education and activism. The purpose of this investigation is to evaluate both the intentions of such projects and the criticism of it through the analysis of a case study at the University of Texas: The Sustainable Design and Development Workshop (SDDW). Through this inquiry, the importance of community engagement to a student's professional development became clear. However, the experience also highlighted the university's inherent responsibility to the citizens with which it is engaging--which requires consideration of issues of funding, timing, accountability, and compromise that is vital to any such project's success. University-community partnerships provide young designers and their educators with important life skills that are not often emphasized within the realms of academia. But how can architects increase the number of people they serve? First they must reassess the service and benefits architecture provides. Defining those is necessary because the greater public-the 98% without access to architects certainly does not understand what architects do, and it is the architect's task, not the public's, to present the reasons that design can help [1]. PMID- 18511396 TI - Impact of odor from industrial hog operations on daily living activities. AB - Intensive industrial animal production systems worldwide require confinement of large numbers of animals in small spaces and concentration of enormous quantities of waste. Industrial hog operations, in particular, have raised public concerns about their adverse impact on public health and sustainable development. Using a community-based participatory research approach and qualitative interviews, we explored people's perception of the impact of odor from these industries on daily living activities as they relate to the beneficial use of property and enjoyment of life. Our research indicates that hog odor limits several leisure time activities and social interactions which could have adverse public health consequences. The results of this study can assist the communities and other stakeholders in public policy development that addresses these concerns. PMID- 18511397 TI - "Doing the heavy lifting: health care workers take back their backs". AB - Health care workers have the highest musculoskeletal disorder prevalence and incidence of any occupational/industry group, and patient handling tasks are so biomechanically demanding that they cannot be made safe through the commonly used, technique-oriented methods such as "back school" training programs. Although there is standard-setting activity for "no-lift" programs in some states, there is still no federal standard. Health care worker unions and nurses' associations have begun to take action through training members in equipment need, use, and acceptance in programs to encourage adoption of no-lifting programs. Acceptance of lifting equipment is increasing due to recognition of the high human and economic costs of MSD, consistent documentation of cost savings from no-lift programs, major improvements in lifting equipment, and shortages of health care staff. An action-oriented training program for health care workers is described that provides knowledge about the 1) Scope of the current problem of back injuries in health care, 2) Costs of injuries, both to workers and to the hospital, 3) Elements of a safe patient-handling program, and 4) Success stories. The program also builds skills through: 1) Hands-on experience with safe lifting equipment, and 2) Assessing organizational and union readiness and planning for action at the workplace. PMID- 18511398 TI - Post-disaster victimization: how survivors of disasters can continue to suffer after the event is over. AB - When public health researchers study the health effects of disasters (whether "naturally-occurring," disasters due to failure of technology, or disasters due to terrorism), some aspects of the post-disaster situation of victims are often overlooked. Social science research has shown that the vast majority of people tend to behave altruistically during and after a disaster. Nevertheless, cases of victimization of survivors do occur. They can include post-disaster victimization of survivors by other individuals (including fellow survivors, opportunistic outsiders, and even unethical aid workers and rogue members of the police, armed forces or international organizations such as the United Nations), groups (such as organized criminal gangs) and institutions (through neglect, incompetence, bureaucratic inefficiency or through institutionalized discriminatory practices). In this article, various kinds of post-disaster victimization that can occur are discussed. PMID- 18511399 TI - The right to know about chemical hazards in Canada, 1982-2006. AB - Traditionally in Canada, there are three health and safety rights: the right to participate (joint workplace health and safety committees); the right to refuse unsafe and unhealthy work; and the right to know about workplace hazards. By the end of the 1970s, the right to know had been established in law across Canada, but it was not enough to cover workplace chemical hazards in particular. The Workplace Hazardous Materials Information System (WHMIS) was a project set up by the Canadian federal government in 1982 to address the issue. This article tells the story of how labor got the progressive WHMIS agreement(1985) and how the agreement has been implemented in the following years. PMID- 18511400 TI - Hooked on a feeling: emotional labor as an occupational hazard of the post industrial age. AB - Emotional labor is a subtle but serious occupational hazard that is likely to spread rapidly as the global service economy continues to grow. Emotional labor requires more than just acting friendly and being helpful to customers; the worker must manage his or her emotions to create a company-dictated experience for customers. The practice of emotional labor in an unsupportive work environment produces work-related stress, which has a wide range of potentially serious health effects. Though many employers do not acknowledge the existence of emotional labor, it is a real occupational hazard that may generate life-altering effects on physical and emotional health. While no official regulations or identification standards specify emotional labor as an occupational hazard, some guidelines exist regarding its outcome: occupational stress. Emotional labor should be recognized as an occupational hazard by the Occupational Safety and Health Administration (OSHA), but this hazard does not lend itself to regulation through standards. The business culture that demands its performance is questioned. PMID- 18511401 TI - 'Protective OSH laws available to only a few'. AB - The American Public Health Association's occupational health and safety section honored Indian safety activist Jagdish Patel last year by giving him its International Award. Here is a brief look at his career followed by his talk at the APHA awards ceremony. PMID- 18511402 TI - Measuring chemicals in people-what would you say? A Boston consensus statement on biomonitoring. PMID- 18511406 TI - A coalescent framework for comparing alternative models of population structure with genetic data: evolution of Celebes toads. AB - Isolation of populations eventually leads to divergence by genetic drift, but if connectivity varies over time, its impact on diversification may be difficult to discern. Even when the habitat patches of multiple species overlap, differences in their demographic parameters, molecular evolution and stochastic events contribute to differences in the magnitude and distribution of their genetic variation. The Indonesian island of Sulawesi, for example, harbours a suite of endemic species whose intraspecific differentiation or interspecific divergence may have been catalysed by habitat fragmentation. To further test this hypothesis, we have performed phylogenetic and coalescent-based analyses on molecular variation in mitochondrial and nuclear DNA of the Celebes toad (Bufo celebensis). Results support a role for habitat fragmentation that led to a population structure in these toads that closely matches distributions of Sulawesi macaque monkeys. Habitat fragmentation, therefore, may also have affected other groups on this island. PMID- 18511407 TI - Transatlantic migration and deep mid-ocean diving by basking shark. AB - Despite being the second largest fish, basking sharks (Cetorhinus maximus) have been assumed to remain in discrete populations. Their known distribution encompasses temperate continental shelf areas, yet until now there has been no evidence for migration across oceans or between hemispheres. Here we present results on the tracks and behaviour of two basking sharks tagged off the British Isles, one of which released its tag off Newfoundland, Canada. During the shark's transit of the North Atlantic, she travelled a horizontal distance of 9589 km and reached a record depth of 1264 m. This result provides the first evidence for a link between European and American populations and indicates that basking sharks make use of deep-water habitats beyond the shelf edge. PMID- 18511408 TI - Characterization of long-term endogenous cardiac repair in children after heart transplantation. AB - AIMS: Circulating cells repopulate the heart at a very low rate in adult humans. The knowledge about time-dependent cardiac regeneration is very limited and the contribution of circulating cells to cardiomyocytes or vascular cells in children is unknown. This study investigates the endogenous repair capacity and the long term incorporation of circulating cells in heart-transplanted children. METHODS AND RESULTS: Cardiac and endothelial chimerism was detected in endomyocardial biopsies of nine children (age 1 months-14 years) with sex-mismatched heart transplantation by fluorescence in situ hybridization. Time from transplantation to biopsy ranged from 1 month up to 10 years. The extent of repopulating cardiomyocytes was 2.39 +/- 1.54% (range: 0-4.2%) and correlated significantly with the time from transplantation to biopsy sampling (r(2) = 0.69, P = 0.006; n = 9). The calculated contribution of male cardiomyocytes in the female heart per year was 0.36 +/- 0.09%. Consistent with the previous reports, the incorporation of vascular cells was higher compared with cardiomyocytes (14.4 +/- 4.17%), but did not correlate in a time-dependent manner. CONCLUSION: Circulating cells contribute to cardiomyocytes and endothelial cells in children after heart transplantation. The incidence of repopulating cardiomyocytes continuously increases in a time-dependent manner ( approximately 4% Y-chromosome(+) cardiomyocytes/10 years) and resembles the cardiac regeneration activity observed in adults. PMID- 18511409 TI - ICAT expression disrupts beta-catenin-TCF interactions and impairs survival of thymocytes and activated mature T cells. AB - T cell factor (TCF) family of transcription factors and beta-catenin critically regulate T cell development as demonstrated by the deletion of the tcf gene, which results in a block early in development that becomes complete in mice bearing tcf/lef double deletion. However, the role of beta-catenin, a major TCF cofactor, remains controversial. To directly address this, we have generated transgenic mice expressing Inhibitor of beta-catenin and TCF (ICAT), a naturally occurring inhibitor that specifically disrupts TCF and beta-catenin interactions. In this report, we demonstrate that disrupting the interaction of beta-catenin with TCF renders adult thymocytes and activated T cells highly susceptible to apoptosis. In contrast to previously reported observations during fetal thymocyte development, these data show that in adult mice, survival and not differentiation of thymocytes, depends on transcription by TCF and beta-catenin. Indeed, we demonstrate that expression of ICAT impedes thymocyte survival by reducing the expression of Bcl(xL) in thymocytes below a critical threshold. Survival of activated mature T cells was also impaired due to diminished expression of activation-induced Bcl(xL). Accordingly, expression of transgenic Bcl-2 rescued activated ICAT-Tg CD4 T cells from apoptosis. Thus, disruption of TCF-beta catenin interactions specifically impairs the survival of thymocytes and activated T cells. PMID- 18511410 TI - Factors associated with marine injuries during vertical chute evacuation training. AB - BACKGROUND: The use of a vertical chute marine evacuation system (MES) is now established on marine vessels; this system has, however, been associated with injuries during training. AIMS: To identify risk factors associated with adverse outcomes during vertical chute MES training and to generate hypotheses for research into reducing injuries. METHODS: Cross-sectional questionnaire study of a cohort of trainees attending vertical chute MES training courses, at two naval training establishments in 2005. Incident data were also collected from both training establishments and from Marine Accident Investigation Branch (MAIB) records. RESULTS: The response rate was 99% (n = 351). Pearson chi square and relative risks (RRs) were calculated for specific outcomes with a number of independent variables. Logistic regression was used to analyse independent variables with a statistically significant (P < 0.05) chi square for the outcome of 'suffering an injury' during training. Injury was associated with body mass index denoting 'overweight', 'obese' or 'morbidly obese' with an odds ratio (OR) of 12.7 (95% CI: 2.2-72.7). Participants below the mean height also had increased risk of suffering an injury during training (OR: 4.8; 95% CI: 1.2-18.5). Phobias and apprehension prior to training increased the risk of injury; reporting of 'some concern over the dark' was statistically significant (OR: 16.6; 95% CI: 1.6 171.5). Life jacket discomfort was also significantly associated with suffering an injury. CONCLUSION: Policy implications include alteration of the MES design and management of risk through training and informed decision making for choosing an evacuation system for ships and ferries. PMID- 18511411 TI - Evolutionary trends in the flowers of Asteridae: is polyandry an alternative to zygomorphy? AB - BACKGROUND AND AIMS: Floral symmetry presents two main states in angiosperms, actinomorphy (polysymmetry or radial symmetry) and zygomorphy (monosymmetry or bilateral symmetry). Transitions from actinomorphy to zygomorphy have occurred repeatedly among flowering plants, possibly in coadaptation with specialized pollinators. In this paper, the rules controlling the evolution of floral symmetry were investigated to determine in which architectural context zygomorphy can evolve. METHODS: Floral traits potentially associated with perianth symmetry shifts in Asteridae, one of the major clades of the core eudicots, were selected: namely the perianth merism, the presence and number of spurs, and the androecium organ number. The evolution of these characters was optimized on a composite tree. Correlations between symmetry and the other morphological traits were then examined using a phylogenetic comparative method. KEY RESULTS: The analyses reveal that the evolution of floral symmetry in Asteridae is conditioned by both androecium organ number and perianth merism and that zygomorphy is a prerequisite to the emergence of spurs. CONCLUSIONS: The statistically significant correlation between perianth zygomorphy and oligandry suggests that the evolution of floral symmetry could be canalized by developmental or spatial constraint. Interestingly, the evolution of polyandry in an actinomorphic context appears as an alternative evolutionary pathway to zygomorphy in Asteridae. These results may be interpreted either in terms of plant-pollinator adaptation or in terms of developmental or physical constraints. The results are discussed in relation to current knowledge about the molecular bases underlying floral symmetry. PMID- 18511412 TI - Embryological features of Tofieldia glutinosa and their bearing on the early diversification of monocotyledonous plants. AB - BACKGROUND AND AIMS: Although much is known about the vegetative traits associated with early monocot evolution, less is known about the reproductive features of early monocotyledonous lineages. A study was made of the embryology of Tofieldia glutinosa, a member of an early divergent monocot clade (Tofieldiaceae), and aspects of its development were compared with the development of other early divergent monocots in order to gain insight into defining reproductive features of early monocots. METHODS: Field-collected developing gynoecial tissues of Tofieldia glutinosa were prepared for histological examination. Over 600 ovules were sectioned and studied using brightfield, differential interference contrast, and fluorescence microscopy. High-resolution digital imaging was used to document important stages of megasporogenesis, megagametogenesis and early endosperm development. KEY RESULTS: Development of the female gametophyte in T. glutinosa is of a modified Polygonum type. At maturity the female gametophyte is seven-celled and 11-nucleate with a standard three-celled egg apparatus, a binucleate central cell (where ultimately, the two polar nuclei will fuse into a diploid secondary nucleus) and three binucleate antipodal cells. The antipodal nuclei persist past fertilization, and the process of double fertilization appears to yield a diploid zygote and triploid primary endosperm cell, as is characteristic of plants with Polygonum type female gametophytes. Endosperm development is helobial, and free-nuclear growth initially proceeds at equal rates in both the micropylar and chalazal endosperm chambers. CONCLUSIONS: The analysis suggests that the shared common ancestor of monocots possessed persistent and proliferating antipodals similar to those found in T. glutinosa and other early-divergent monocots (e.g. Acorus and members of the Araceae). Helobial endosperm among monocots evolved once in the common ancestor of all monocots excluding Acorus. Thus, the analysis further suggests that helobial endosperm in monocots is homoplasious with those helobial endosperms that are present in water lilies and eudicot angiosperms. PMID- 18511413 TI - Efflux-mediated response of Staphylococcus aureus exposed to ethidium bromide. AB - OBJECTIVES: By adapting an antibiotic-susceptible Staphylococcus aureus strain to increasing concentrations of ethidium bromide, a known substrate of efflux pumps (EPs), and by phenotypically and genotypically analysing the resulting progeny, we characterized the molecular mechanisms of S. aureus adaptation to ethidium bromide. METHODS: S. aureus ATCC 25923 was grown in increasing concentrations of ethidium bromide. The MICs of representatives of eight classes of antibiotics, eight biocides and two dyes against ATCC 25923 and its ethidium bromide-resistant progeny ATCC 25923(EtBr) were determined with or without six efflux pump inhibitors (EPIs). Efflux activity in the presence/absence of EPIs was evaluated by real-time fluorometry. The presence and expression of eight EP genes were assayed by PCR and quantitative RT-PCR (qRT-PCR), respectively. Mutations in grlA, gyrA and norA promoter regions were screened by DNA sequencing. RESULTS: Compared with its parental strain, ATCC 25923(EtBr) was 32-fold more resistant to ethidium bromide and also more resistant to biocides and hydrophilic fluoroquinolones. Resistance to these could be reduced by the EPIs chlorpromazine, thioridazine and reserpine. Increased efflux of ethidium bromide by ATCC 25923(EtBr) could be inhibited by the same EPIs. qRT-PCR showed that norA was 35-fold over-expressed in ATCC 25923(EtBr), whereas the remaining EP genes showed no significant increase in their expression. Sequencing of the norA promoter region revealed a 70 bp deletion in ATCC 25923(EtBr). CONCLUSIONS: Exposure of S. aureus to quaternary compounds such as ethidium bromide results in decreased susceptibility of the organism to a wide variety of compounds, including quinolones and biocides through an efflux-mediated response, which for strain ATCC 25923 is mainly NorA-mediated. This altered expression may result from alterations in the norA promoter region. PMID- 18511414 TI - Site-specific androgen receptor serine phosphorylation linked to epidermal growth factor-dependent growth of castration-recurrent prostate cancer. AB - The androgen receptor (AR) is required for prostate cancer development and contributes to tumor progression after remission in response to androgen deprivation therapy. Epidermal growth factor (EGF) increases AR transcriptional activity at low levels of androgen in the CWR-R1 prostate cancer cell line derived from the castration-recurrent CWR22 prostate cancer xenograft. Here we report that knockdown of AR decreases EGF stimulation of prostate cancer cell growth and demonstrate a mechanistic link between EGF and AR signaling. The EGF induced increase in AR transcriptional activity is dependent on phosphorylation at mitogen-activated protein kinase consensus site Ser-515 in the AR NH(2) terminal region and at protein kinase C consensus site Ser-578 in the AR DNA binding domain. Phosphorylation at these sites alters the nuclear-cytoplasmic shuttling of AR and AR interaction with the Ku-70/80 regulatory subunits of DNA dependent protein kinase. Abolishing AR Ser-578 phosphorylation by introducing an S578A mutation eliminates the AR transcriptional response to EGF and increases both AR binding of Ku-70/80 and nuclear retention of AR in association with hyperphosphorylation of AR Ser-515. The results support a model in which AR transcriptional activity increases castration-recurrent prostate cancer cell growth in response to EGF by site-specific serine phosphorylation that regulates nuclear-cytoplasmic shuttling through interactions with the Ku-70/80 regulatory complex. PMID- 18511415 TI - Structural consequences of nucleophosmin mutations in acute myeloid leukemia. AB - Mutations affecting NPM1 (nucleophosmin) are the most common genetic lesions found in acute myeloid leukemia (AML). NPM1 is one of the most abundant proteins found in the nucleolus and has links to the MDM2/p53 tumor suppressor pathway. A distinctive feature of NPM1 mutants in AML is their aberrant localization to the cytoplasm of leukemic cells. This mutant phenotype is the result of the substitution of several C-terminal residues, including one or two conserved tryptophan residues, with a leucine-rich nuclear export signal. The exact molecular mechanism underlying the loss of nucleolar retention, and the role of the tryptophans, remains unknown. In this study we have determined the structure of an independently folded globular domain in the C terminus of NPM1 using NMR spectroscopy, and we report that the conserved tryptophans are critical for structure. This domain is necessary for the nucleolar targeting of NPM1 and is disrupted by mutations in AML with cytoplasmic NPM1. Furthermore, we identify conserved surface-exposed lysine residues that are functionally rather than structurally important for nucleolar localization. This study provides new focus for efforts to understand the pathogenesis of AML with cytoplasmic NPM1 and may be used to aid the design of small molecules that target the C-terminal domain of NPM1 to act as novel anti-proliferative and anti-leukemia therapeutics. PMID- 18511416 TI - Acetylcholinesterase associates differently with its anchoring proteins ColQ and PRiMA. AB - Acetylcholinesterase tetramers are inserted in the basal lamina of neuromuscular junctions or anchored in cell membranes through the interaction of four C terminal t peptides with proline-rich attachment domains (PRADs) of cholinesterase-associated collagen Q (ColQ) or of the transmembrane protein PRiMA (proline-rich membrane anchor). ColQ and PRiMA differ in the length of their proline-rich motifs (10 and 15 residues, respectively). ColQ has two cysteines upstream of the PRAD, which are disulfide-linked to two AChE(T) subunits ("heavy" dimer), and the other two subunits are disulfide-linked together ("light" dimer). In contrast, PRiMA has four cysteines upstream of the PRAD. We examined whether these cysteines could be linked to AChE(T) subunits in complexes formed with PRiMA in transfected COS cells and in the mammalian brain. For comparison, we studied complexes formed with N-terminal fragments of ColQ, N-terminal fragments of PRiMA, and chimeras in which the upstream regions containing the cysteines were exchanged. We also compared the effect of mutations in the t peptides on their association with the two PRADs. We report that the two PRADs differ in their interaction with AChE(T) subunits; in complexes formed with the PRAD of PRiMA, we observed light dimers, but very few heavy dimers, even though such dimers were formed with the PQ chimera in which the N-terminal region of PRiMA was associated with the PRAD of ColQ. Complexes with PQ or with PRiMA contained heavy components, which migrated abnormally in SDS-PAGE but probably resulted from disulfide bonding of four AChE(T) subunits with the four upstream cysteines of the associated protein. PMID- 18511417 TI - Substrate recognition of VAMP-2 by botulinum neurotoxin B and tetanus neurotoxin. AB - Botulinum neurotoxin (BoNT; serotypes A-G) and tetanus neurotoxin elicit flaccid and spastic paralysis, respectively. These neurotoxins are zinc proteases that cleave SNARE proteins to inhibit synaptic vesicle fusion to the plasma membrane. Although BoNT/B and tetanus neurotoxin (TeNT) cleave VAMP-2 at the same scissile bond, their mechanism(s) of VAMP-2 recognition is not clear. Mapping experiments showed that residues 60-87 of VAMP-2 were sufficient for efficient cleavage by BoNT/B and that residues 40-87 of VAMP-2 were sufficient for efficient TeNT cleavage. Alanine-scanning mutagenesis and kinetic analysis identified three regions within VAMP-2 that were recognized by BoNT/B and TeNT: residues adjacent to the site of scissile bond cleavage (cleavage region) and residues located within N-terminal and C-terminal regions relative to the cleavage region. Analysis of residues within the cleavage region showed that mutations at the P7, P4, P2, and P1' residues of VAMP-2 had the greatest inhibition of LC/B cleavage (> or =32-fold), whereas mutations at P7, P4, P1', and P2' residues of VAMP-2 had the greatest inhibition of LC/TeNT cleavage (> or =64-fold). Residues within the cleavage region influenced catalysis, whereas residues N-terminal and C-terminal to the cleavage region influenced binding affinity. Thus, BoNT/B and TeNT possess similar organization but have unique residues to recognize and cleave VAMP-2. These studies provide new insights into how the clostridial neurotoxins recognize their substrates. PMID- 18511418 TI - Substrate recognition mechanism of VAMP/synaptobrevin-cleaving clostridial neurotoxins. AB - Botulinum neurotoxins (BoNTs) and tetanus neurotoxin (TeNT) inhibit neurotransmitter release by proteolyzing a single peptide bond in one of the three soluble N-ethylmaleimide-sensitive factor attachment protein receptors SNAP 25, syntaxin, and vesicle-associated membrane protein (VAMP)/synaptobrevin. TeNT and BoNT/B, D, F, and G of the seven known BoNTs cleave the synaptic vesicle protein VAMP/synaptobrevin. Except for BoNT/B and TeNT, they cleave unique peptide bonds, and prior work suggested that different substrate segments are required for the interaction of each toxin. Although the mode of SNAP-25 cleavage by BoNT/A and E has recently been studied in detail, the mechanism of VAMP/synaptobrevin proteolysis is fragmentary. Here, we report the determination of all substrate residues that are involved in the interaction with BoNT/B, D, and F and TeNT by means of systematic mutagenesis of VAMP/synaptobrevin. For each of the toxins, three or more residues clustered at an N-terminal site remote from the respective scissile bond are identified that affect solely substrate binding. These exosites exhibit different sizes and distances to the scissile peptide bonds for each neurotoxin. Substrate segments C-terminal of the cleavage site (P4 P4') do not play a role in the catalytic process. Mutation of residues in the proximity of the scissile bond exclusively affects the turnover number; however, the importance of individual positions at the cleavage sites varied for each toxin. The data show that, similar to the SNAP-25 proteolyzing BoNT/A and E, VAMP/synaptobrevin-specific clostridial neurotoxins also initiate substrate interaction, employing an exosite located N-terminal of the scissile peptide bond. PMID- 18511419 TI - Modulation of nicotinic acetylcholine receptor conformational state by free fatty acids and steroids. AB - Steroids and free fatty acids (FFA) are noncompetitive antagonists of the nicotinic acetylcholine receptor (AChR). Their site of action is purportedly located at the lipid-AChR interface, but their exact mechanism of action is still unknown. Here we studied the effect of structurally different FFA and steroids on the conformational equilibrium of the AChR in Torpedo californica receptor-rich membranes. We took advantage of the higher affinity of the fluorescent AChR open channel blocker, crystal violet, for the desensitized state than for the resting state. Increasing concentrations of steroids and FFA decreased the K(D) of crystal violet in the absence of agonist; however, only cis-unsaturated FFA caused an increase in K(D) in the presence of agonist. This latter effect was also observed with treatments that caused the opposite effects on membrane polarity, such as phospholipase A(2) treatment or temperature increase (decreasing or increasing membrane polarity, respectively). Quenching by spin labeled fatty acids of pyrene-labeled AChR reconstituted into model membranes, with the label located at the gammaM4 transmembrane segment, disclosed the occurrence of conformational changes induced by steroids and cis-unsaturated FFA. The present work is a step forward in understanding the mechanism of action of this type of molecules, suggesting that the direct contact between exogenous lipids and the AChR transmembrane segments removes the AChR from its resting state and that membrane polarity modulates the AChR activation equilibrium by an independent mechanism. PMID- 18511420 TI - G2E3 is a dual function ubiquitin ligase required for early embryonic development. AB - G2E3 is a putative ubiquitin ligase (E3) identified in a microarray screen for mitotic regulatory proteins. It shuttles between the cytoplasm and nucleus, concentrating in nucleoli and relocalizing to the nucleoplasm in response to DNA damage. In this study, we demonstrate that G2E3 is an unusual ubiquitin ligase that is essential in early embryonic development to prevent apoptotic death. This protein has a catalytically inactive HECT domain and two distinct RING-like ubiquitin ligase domains that catalyze lysine 48-linked polyubiquitination. To address in vivo function, we generated a knock-out mouse model of G2E3 deficiency that incorporates a beta-galactosidase reporter gene under control of the endogenous promoter. Animals heterozygous for G2E3 inactivation are phenotypically normal with no overt change in development, growth, longevity, or fertility, whereas G2E3 null embryos die prior to implantation. Although normal numbers of G2E3(-/-) blastocysts are present at embryonic day 3.5, these blastocysts involute in culture as a result of massive apoptosis. Using beta galactosidase staining as a marker for protein expression, we demonstrate that G2E3 is predominantly expressed within the central nervous system and the early stages of limb bud formation of the developing embryo. In adult animals, the most intense staining is found in Purkinje cell bodies and cells lining the ductus deferens. In summary, G2E3 is a dual function ubiquitin ligase essential for prevention of apoptosis in early embryogenesis. PMID- 18511421 TI - Active-site mapping of a Populus xyloglucan endo-transglycosylase with a library of xylogluco-oligosaccharides. AB - Restructuring the network of xyloglucan (XG) and cellulose during plant cell wall morphogenesis involves the action of xyloglucan endo-transglycosylases (XETs). They cleave the XG chains and transfer the enzyme-bound XG fragment to another XG molecule, thus allowing transient loosening of the cell wall and also incorporation of nascent XG during expansion. The substrate specificity of a XET from Populus (PttXET16-34) has been analyzed by mapping the enzyme binding site with a library of xylogluco-oligosaccharides as donor substrates using a labeled heptasaccharide as acceptor. The extended binding cleft of the enzyme is composed of four negative and three positive subsites (with the catalytic residues between subsites -1 and +1). Donor binding is dominated by the higher affinity of the XXXG moiety (G=Glcbeta(1-->4) and X=Xylalpha(1-->6)Glcbeta(1-->4)) of the substrate for positive subsites, whereas negative subsites have a more relaxed specificity, able to bind (and transfer to the acceptor) a cello-oligosaccharyl moiety of hybrid substrates such as GGGGXXXG. Subsite mapping with k(cat)/K(m) values for the donor substrates showed that a GG-unit on negative and -XXG on positive subsites are the minimal requirements for activity. Subsites -2 and -3 (for backbone Glc residues) and +2' (for Xyl substitution at Glc in subsite +2) have the largest contribution to transition state stabilization. GalGXXXGXXXG (Gal=Galbeta(1-->4)) is the best donor substrate with a "blocked" nonreducing end that prevents polymerization reactions and yields a single transglycosylation product. Its kinetics have unambiguously established that the enzyme operates by a ping-pong mechanism with competitive inhibition by the acceptor. PMID- 18511422 TI - RNA primer handoff in bacteriophage T4 DNA replication: the role of single stranded DNA-binding protein and polymerase accessory proteins. AB - In T4 phage, coordinated leading and lagging strand DNA synthesis is carried out by an eight-protein complex termed the replisome. The control of lagging strand DNA synthesis depends on a highly dynamic replisome with several proteins entering and leaving during DNA replication. Here we examine the role of single stranded binding protein (gp32) in the repetitive cycles of lagging strand synthesis. Removal of the protein-interacting domain of gp32 results in a reduction in the number of primers synthesized and in the efficiency of primer transfer to the polymerase. We find that the primase protein is moderately processive, and this processivity depends on the presence of full-length gp32 at the replication fork. Surprisingly, we find that an increase in the efficiency of primer transfer to the clamp protein correlates with a decrease in the dissociation rate of the primase from the replisome. These findings result in a revised model of lagging strand DNA synthesis where the primase remains as part of the replisome after each successful cycle of Okazaki fragment synthesis. A delay in primer transfer results in an increased probability of the primase dissociating from the replication fork. The interplay between gp32, primase, clamp, and clamp loader dictates the rate and efficiency of primer synthesis, polymerase recycling, and primer transfer to the polymerase. PMID- 18511423 TI - Protease-activated receptor-2 exerts contrasting model-specific effects on acute experimental pancreatitis. AB - Protease-activated receptor-2 (PAR2) is a 7-transmembrane G-protein-coupled tethered ligand receptor that is expressed by pancreatic acinar and ductal cells. It can be physiologically activated by trypsin. Previously reported studies (Namkung, W., Han, W., Luo, X., Muallem, S., Cho, K. H., Kim, K. H., and Lee, M. G. (2004) Gastroenterology 126, 1844-1859; Sharma, A., Tao, X., Gopal, A., Ligon, B., Andrade-Gordon, P., Steer, M. L., and Perides, G. (2005) Am. J. Physiol. 288, G388-G395) have shown that PAR2 activation exerts a protective effect on the experimental model of pancreatitis induced by supramaximal secretagogue (caerulein) stimulation. We now show that PAR2 exerts a worsening effect on a different model of experimental pancreatitis, i.e. one induced by retrograde pancreatic ductal infusion of bile salts. In vitro studies using freshly prepared pancreatic acini show that genetic deletion of PAR2 reduces bile salt-induced pathological calcium transients, acinar cell injury, and activation of c-Jun N terminal kinase, whereas genetic deletion of PAR2 has the opposite or no effect on these pancreatitis-related events when they are elicited, in vitro, by caerulein stimulation. Studies employing a combination of trypsin inhibition and activation of PAR2 with the activating peptide SLIGRL show that all these differences indeed depend on the activation of PAR2. These studies are the first to report that a single perturbation can have model-specific and opposite effects on pancreatitis, and they underscore the importance of performing mechanistic pancreatitis studies using two dissimilar models of the disease to detect idiosyncratic, model-specific events. We suggest PAR2 activation exerts a worsening effect on the severity of clinical pancreatitis and that interventions interfering with PAR2 activation may be of benefit in the treatment of patients with severe pancreatitis. PMID- 18511424 TI - Macrophage secretory phospholipase A2 group X enhances anti-inflammatory responses, promotes lipid accumulation, and contributes to aberrant lung pathology. AB - Secreted phospholipase A2 group X (sPLA(2)-X) is one of the most potent enzymes of the phospholipase A(2) lipolytic enzyme superfamily. Its high catalytic activity toward phosphatidylcholine (PC), the major phospholipid of cell membranes and low-density lipoproteins (LDL), has implicated sPLA(2)-X in chronic inflammatory conditions such as atherogenesis. We studied the role of sPLA(2)-X enzyme activity in vitro and in vivo, by generating sPLA(2)-X-overexpressing macrophages and transgenic macrophage-specific sPLA(2)-X mice. Our results show that sPLA(2)-X expression inhibits macrophage activation and inflammatory responses upon stimulation, characterized by reduced cell adhesion and nitric oxide production, a decrease in tumor necrosis factor (TNF), and an increase in interleukin (IL)-10. These effects were mediated by an increase in IL-6, and enhanced production of prostaglandin E(2) (PGE(2)) and 15-deoxy-Delta12,14 prostaglandin J(2) (PGJ(2)). Moreover, we found that overexpression of active sPLA(2)-X in macrophages strongly increases foam cell formation upon incubation with native LDL but also oxidized LDL (oxLDL), which is mediated by enhanced expression of scavenger receptor CD36. Transgenic sPLA(2)-X mice died neonatally because of severe lung pathology characterized by interstitial pneumonia with massive granulocyte and surfactant-laden macrophage infiltration. We conclude that overexpression of the active sPLA(2)-X enzyme results in enhanced foam cell formation but reduced activation and inflammatory responses in macrophages in vitro. Interestingly, enhanced sPLA(2)-X activity in macrophages in vivo leads to fatal pulmonary defects, suggesting a crucial role for sPLA(2)-X in inflammatory lung disease. PMID- 18511425 TI - The impact of founder events on chromosomal variability in multiply mating species. AB - In species with heterogametic males, the relative levels of X chromosome versus autosome diversity hold key information about the evolutionary forces at work in a population. It has been shown that population size changes alter the ratio of X linked to autosomal (X/A) variation, with population size reductions and recent bottlenecks leading to decreased X/A diversity ratios. Here we use theory and simulation to investigate a separate demographic effect-that of founder events involving multiply mated females-and find that it leads to much stronger reductions in X/A diversity ratios than are produced by simple population size changes. Investigating the potential of this process to account for sharply reduced X-linked diversity in European Drosophila melanogaster, we find that this model yields predictions that are compatible with the empirical data. PMID- 18511426 TI - A maximum likelihood method for detecting directional evolution in protein sequences and its application to influenza A virus. AB - We develop a model-based phylogenetic maximum likelihood test for evidence of preferential substitution toward a given residue at individual positions of a protein alignment--directional evolution of protein sequences (DEPS). DEPS can identify both the target residue and sites evolving toward it, help detect selective sweeps and frequency-dependent selection--scenarios that confound most existing tests for selection, and achieve good power and accuracy on simulated data. We applied DEPS to alignments representing different genomic regions of influenza A virus (IAV), sampled from avian hosts (H5N1 serotype) and human hosts (H3N2 serotype), and identified multiple directionally evolving sites in 5/8 genomic segments of H5N1 and H3N2 IAV. We propose a simple descriptive classification of directionally evolving sites into 5 groups based on the temporal distribution of residue frequencies and document known functional correlates, such as immune escape or host adaptation. PMID- 18511427 TI - Perinatal factors and the risk of asthma in childhood--a population-based register study in Finland. AB - The aim of the study was to assess whether perinatal factors are associated with the risk of asthma in childhood in a register-based, nested case-control study in Finland. All children born between January 1, 1996, and April 30, 2004, who were entitled to a special reimbursement for antiasthmatic drugs (i.e., had diagnosed asthma by 2006 and had purchased inhaled corticosteroids or montelukast at least once), were identified (n = 21,038). For each case, one matched control child was selected. The associations between perinatal factors, derived from the Finnish Medical Birth Register, and the risk of asthma were analyzed by conditional logistic regression. In the final multivariate model, maternal asthma, young age, smoking, previous miscarriages, and a high number of previous deliveries, as well as cesarean section, low gestational age, and low ponderal index, were associated with an increased risk of asthma in children diagnosed before the age of 3 years. Among children diagnosed at the age of 3 years or later, maternal asthma, low gestational age, and low ponderal index were associated with an increased risk, and a high number of previous deliveries was associated with a decreased risk of asthma. In conclusion, perinatal factors play a role in the development of asthma in childhood, but the etiology may differ in early and late-onset asthma. PMID- 18511428 TI - On the estimation of additive interaction by use of the four-by-two table and beyond. AB - A four-by-two table with its four rows representing the presence and absence of gene and environmental factors has been suggested as the fundamental unit in the assessment of gene-environment interaction. For such a table to be more meaningful from a public health perspective, it is important to estimate additive interaction. A confidence interval procedure proposed by Hosmer and Lemeshow has become widespread. This article first reveals that the Hosmer-Lemeshow procedure makes an assumption that confidence intervals for risk ratios are symmetric and then presents an alternative that uses the conventional asymmetric intervals for risk ratios to set confidence limits for measures of additive interaction. For the four-by-two table, the calculation involved requires no statistical programs but only elementary calculations. Simulation results demonstrate that this new approach can perform almost as well as the bootstrap. Corresponding calculations in more complicated situations can be simplified by use of routine output from multiple regression programs. The approach is illustrated with three examples. A Microsoft Excel spreadsheet and SAS codes for the calculations are available from the author and the Journal's website, respectively. PMID- 18511429 TI - MRP2 and the DMPS- and DMSA-mediated elimination of mercury in TR(-) and control rats exposed to thiol S-conjugates of inorganic mercury. AB - Cysteine (Cys) and homocysteine (Hcy)-S-conjugates of inorganic mercury (Hg2+) are transportable species of Hg2+ that are taken up readily by proximal tubular cells. The metal chelators, 2,3-dimercaptopropane-1-sulfonic acid (DMPS) and meso 2,3-dimercaptosuccinic acid (DMSA), have been used successfully to extract Hg2+ from these cells, presumably via the multidrug resistance protein (Mrp2). In the current study, we tested the hypothesis that Mrp2 is involved in the DMPS- and DMSA-mediated extraction of Hg2+ following administration of Hg2+ as an S conjugate of Cys or Hcy. To test this hypothesis, control and TR(-) (Mrp2 deficient) rats were injected with 0.5 micromol/kg HgCl2 (containing 203Hg2+) conjugated to 1.25 micromol/kg Cys or Hcy. After 24 and 28 h, rats were treated with saline or 100 mg/kg DMPS or DMSA. Tissues were harvested 48 h after Hg2+ exposure. The renal and hepatic burden of Hg2+ was greater in saline-injected TR- rats than in corresponding controls. Accordingly, the content of Hg2+ in the urine and feces was less in TR- rats than in controls. Following treatment with DMPS or DMSA, the renal content of Hg2+ in both groups of rats was reduced significantly and the urinary excretion of Hg2+ was increased. In liver, the effect of each chelator appeared to be dependent upon the form in which Hg2+ was administered. In vitro experiments provide direct evidence indicating that DMPS and DMSA-S-conjugates of Hg2+ are substrates for Mrp2. Overall, these data support our hypothesis that Mrp2 is involved in the DMPS and DMSA-mediated extraction of the body burden of Hg2+. PMID- 18511430 TI - Decreased urinary beta-defensin-1 expression as a biomarker of response to arsenic. AB - Ingestion of arsenic (As) through contaminated drinking water results in increased risks of skin, lung, kidney, and bladder cancers. Due to its association with kidney and bladder cancers, we hypothesized that analysis of the urinary proteome could provide insight into the mechanisms of As toxicity. Urine from participants in a cross-sectional As biomarker study conducted in Nevada, classified as having either high (>or= 100 microg total urinary As/l) or low exposure (< 100 microg total urinary As/l) was analyzed by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Two polypeptides, 2.21 and 4.37 kDa, were significantly decreased in the high exposure group (p < 0.05) and were limited to men when stratified by sex. To replicate these findings, urine from participants in a second As study in Chile was analyzed and results confirmed the decrease of the 4.37 kDa polypeptide as well as a 4.76 kDa polypeptide among highly exposed men. These peaks were identified and confirmed as human beta-defensin-1 (HBD-1) peptides. In a separate in vitro experiment, gene expression analysis of As-treated cell lines demonstrated reduced HBD1 mRNA confirming that the observed decrease in HBD-1 resulted from As exposure. HBD-1 is an antimicrobial peptide constitutively expressed in multiple tissues including epithelial cells of the respiratory and urogenital systems. Recent studies support its role as a tumor suppressor gene for urological cancers suggesting that decreased HBD-1 levels may play a role in the development of cancers associated with As exposure. Further studies are warranted to investigate the role of HBD-1 in As-related toxicity. PMID- 18511431 TI - Effects of perfluorobutyrate exposure during pregnancy in the mouse. AB - Perfluorobutyrate (PFBA) is a perfluoroalkyl acid (PFAA) found in the environment. Previous studies have indicated developmental toxicity of PFAAs (perfluorooctane sulfonate [PFOS] and perfluorooctanoate [PFOA]); the current study examines that of PFBA. PFBA/NH4(+) was given to timed-pregnant CD-1 mice by oral gavage daily from gestational day (GD) 1 to 17 at 35, 175, or 350 mg/kg (chosen to approximate the developmentally toxic doses of PFOA); controls received water. At GD 18, serum levels of PFBA were 3.8, 4.4, and 2.5 microg/ml, respectively, in the three treated groups. PFBA did not significantly affect maternal weight gain, number of implantations, fetal viability, fetus weight, or incidence of fetal malformations. Incidence of full-litter loss was significantly greater in the 350 mg/kg group, and maternal liver weights were significantly increased in the 175 and 350 mg/kg groups. In contrast to PFOA and PFOS, PFBA exposure during pregnancy did not adversely affect neonatal survival or postnatal growth. Liver enlargement was detected in the PFBA-exposed pups on postnatal day (PD) 1, but not by PD 10. Expression of selected hepatic genes in PFBA-exposed pups at PD 1 did not reveal any significant changes from controls. A significant delay in eye-opening in offspring was detected in all three PFBA groups, and slight delays in the onset of puberty were noted in the 175 and 350 mg/kg groups. These data suggest that exposure to PFBA during pregnancy in the mouse did not produce developmental toxicity comparable to that observed with PFOA, in part, due to rapid elimination of the chemical. PMID- 18511432 TI - MicroRNAs: novel regulators in cardiac development and disease. AB - MicroRNAs (miRNAs) are endogenous, small ribonucleotides regulating the translation of target messenger RNAs that have been shown to be involved in orchestrating growth, development, function, and stress responses of various organs, including the heart. Muscle miRNAs are mainly controlled by a network of myogenic transcription factors, and throughout cardiac development they fine-tune regulatory protein levels in a spatiotemporal manner. Recent profiling studies revealed that miRNA expression patterns are derailed in both human cardiac disease and animal models of cardiac hypertrophy and failure. Modulation of miRNA expression in vitro as well as in vivo has revealed an important role of miRNAs in regulating heart function, particularly cardiac growth and conductance. Here, we overview the recent findings on miRNAs in cardiac development and disease and report the latest advances in the identification and validation of miRNA targets, which are important for a comprehensive understanding of cardiac miRNA function. Finally, we focus on the development and use of miRNA antagonists (antagomirs) to target miRNAs in vivo, which may translate into novel therapeutic strategies for heart disease in the future. PMID- 18511433 TI - A review of methods for assessment of coronary microvascular disease in both clinical and experimental settings. AB - Obstructive disease of the large coronary arteries is the prominent cause for angina pectoris. However, angina may also occur in the absence of significant coronary atherosclerosis or coronary artery spasm, especially in women. Myocardial ischaemia in these patients is often associated with abnormalities of the coronary microcirculation and may thus represent a manifestation of coronary microvascular disease (CMD). Elucidation of the role of the microvasculature in the genesis of myocardial ischaemia and cardiac damage-in the presence or absence of obstructive coronary atherosclerosis-will certainly result in more rational diagnostic and therapeutic interventions for patients with ischaemic heart disease. Specifically targeted research based on improved assessment modalities is needed to improve the diagnosis of CMD and to translate current molecular, cellular, and physiological knowledge into new therapeutic options. PMID- 18511434 TI - AP-1 and STAT-1 decoy oligodeoxynucleotides attenuate transplant vasculopathy in rat cardiac allografts. AB - AIMS: Cardiac allograft vasculopathy (CAV) continues to be an unsolved clinical problem requiring the development of new therapeutic strategies. We have previously demonstrated that ex vivo donor allograft treatment with decoy oligodeoxynucleotides (ODN) targeting the transcription factors, activator protein-1 (AP-1) or signal transducer and activator of transcription-1 (STAT-1), delays acute rejection and prolongs cardiac allograft survival. Here, we investigated whether this treatment regime also prevents the occurrence of CAV in a fully allogeneic rat heart transplantation model. METHODS AND RESULTS: Wistar Furth rat cardiac allografts were perfused ex vivo with AP-1 decoy ODN, STAT-1 decoy ODN, or buffer solution and transplanted into the abdomen of Lewis rats immunosuppressed with cyclosporine. Treatment with both decoy ODNs but not vehicle significantly attenuated the incidence and severity of CAV. Laser assisted microdissection/real-time polymerase chain reaction as well as immunohistochemistry analyses revealed a significant increase in CD40 abundance in the coronary endothelial cells and medial smooth muscle cells on day 1 post transplantation which was virtually abolished upon AP-1 or STAT-1 decoy ODN treatment. While the AP-1 decoy ODN primarily attenuated basal CD40 expression, the STAT-1 decoy ODN suppressed tumour necrosis factor-alpha-/interferon-gamma stimulated expression of CD40 in rat native endothelial cells. CONCLUSION: Treating donor hearts with decoy ODNs neutralizing AP-1 or STAT-1 at the time of transplantation prevents upregulation of CD40 expression in the graft coronary arteries and effectively inhibits CAV. PMID- 18511435 TI - Genome structure of the legume, Lotus japonicus. AB - The legume Lotus japonicus has been widely used as a model system to investigate the genetic background of legume-specific phenomena such as symbiotic nitrogen fixation. Here, we report structural features of the L. japonicus genome. The 315.1-Mb sequences determined in this and previous studies correspond to 67% of the genome (472 Mb), and are likely to cover 91.3% of the gene space. Linkage mapping anchored 130-Mb sequences onto the six linkage groups. A total of 10,951 complete and 19,848 partial structures of protein-encoding genes were assigned to the genome. Comparative analysis of these genes revealed the expansion of several functional domains and gene families that are characteristic of L. japonicus. Synteny analysis detected traces of whole-genome duplication and the presence of synteny blocks with other plant genomes to various degrees. This study provides the first opportunity to look into the complex and unique genetic system of legumes. PMID- 18511436 TI - Soybean seed extracts preferentially express genomic loci of Bradyrhizobium japonicum in the initial interaction with soybean, Glycine max (L.) Merr. AB - Initial interaction between rhizobia and legumes actually starts via encounters of both partners in the rhizosphere. In this study, the global expression profiles of Bradyrhizobium japonicum USDA 110 in response to soybean (Glycine max) seed extracts (SSE) and genistein, a major soybean-released isoflavone for nod genes induction of B. japonicum, were compared. SSE induced many genomic loci as compared with genistein (5.0 microM), nevertheless SSE-supplemented medium contained 4.7 microM genistein. SSE markedly induced four predominant genomic regions within a large symbiosis island (681 kb), which include tts genes (type III secretion system) and various nod genes. In addition, SSE-treated cells expressed many genomic loci containing genes for polygalacturonase (cell-wall degradation), exopolysaccharide synthesis, 1-aminocyclopropane-1-carboxylate deaminase, ribosome proteins family and energy metabolism even outside symbiosis island. On the other hand, genistein-treated cells exclusively showed one expression cluster including common nod gene operon within symbiosis island and six expression loci including multidrug resistance, which were shared with SSE treated cells. Twelve putatively regulated genes were indeed validated by quantitative RT-PCR. Several SSE-induced genomic loci likely participate in the initial interaction with legumes. Thus, these results can provide a basic knowledge for screening novel genes relevant to the B. japonicum- soybean symbiosis. PMID- 18511437 TI - Computed tomography-fluoroscopy overlay evaluation during catheter ablation of left atrial arrhythmia. AB - AIMS: Proper visualization of left atrial (LA) and pulmonary vein (PV) anatomy is of crucial importance during atrial fibrillation (AF) ablation. This two-centre study evaluated a new automatic computed tomography (CT)-fluoroscopy overlay system (EP navigator, Philips Medical Systems, Best, The Netherlands) and the accuracy of different registration methods. METHODS AND RESULTS: Fifty-six consecutive patients (age: 56 +/- 14) with symptomatic AF underwent contrast CT of the LA/PV prior to ablation. Three registration methods were evaluated and validated by comparison with LA angiography: (i) catheter registration: the placement of catheters in identifiable anatomical structures; (ii) heart contour: based on aligning the fluoroscopy heart contours and the 3D-rendered CT volume; and (iii) spine registration: based on automatically aligning the segmented CT spine on fluoroscopy. Computed tomography segmentation was achieved in all but one patient due to motion artefacts. The mean duration of segmentation was 10 min and average registration lasted 7 min. Catheter and heart contour registration were highly accurate (discrepancy of 1.3 +/- 0.6 and 0.3 +/- 0.5 mm, respectively) when compared with spine registration (17 +/- 9 mm, P < 0.05). The EP navigator was helpful during trans-septal puncture, gave an internal view of the atria and allowed tracking of ablation lesions. CONCLUSION: The EP navigator enabled accurate live integration of CT images and real-time fluoroscopy. Registration utilizing catheter placement or heart contours was stable and reliable. PMID- 18511438 TI - Benefit of cardiac resynchronization therapy in atrial fibrillation patients vs. patients in sinus rhythm: the role of atrioventricular junction ablation. AB - AIMS: To assess the clinical benefit of cardiac resynchronization therapy (CRT) in patients with atrial fibrillation (AF) compared with patients in sinus rhythm (SR), and to evaluate the impact of atrioventricular junction (AVJ) ablation on the outcome of AF patients undergoing CRT. METHODS AND RESULTS: We conducted a retrospective analysis of 131 consecutive heart failure (HF) patients who underwent CRT implantation. Three groups were considered: SR (n = 78), AF with AVJ ablation (n = 26), and AF without AVJ ablation (n = 27). Patients were evaluated for the occurrence of cardiac death, hospitalization for HF, and responsiveness to CRT (survival with improvement of >or=1 New York Heart Association class at 6 months). The three groups showed a significant improvement in functional class. However, the proportion of responders was significantly lower in AF patients without AVJ ablation (52 vs. 79% in SR and 85% in AF with AVJ ablation, P < 0.008). Atrial fibrillation without AVJ ablation was also independently associated with mortality (HR 5.22, 95% CI: 1.60-17.01, P = 0.006) and hospitalization for HF during the first 12 months (HR 6.23, 95% CI: 2.09 18.54, P = 0.001). The outcomes of AF with AVJ ablation patients were similar to the outcomes of patients in SR. CONCLUSION: Sinus rhythm and AF patients display similar survival and clinical improvement after CRT implantation, provided that AVJ ablation is performed in the latter. PMID- 18511439 TI - Influence of systolic-pressure-variation-guided intraoperative fluid management on organ function and oxygen transport. AB - BACKGROUND: Dynamic variables, for example, systolic pressure variation (SPV), are superior to filling pressures for assessing fluid responsiveness. We analysed the effects of SPV-guided intraoperative fluid management on organ function and perfusion when compared with routine care. METHODS: Eighty patients (44 female and 36 male) undergoing elective major abdominal surgery were randomly assigned to a control group [n=40, mean age 66 (sd 10), range 40-84 yr] or SPV group [n=40, age 61 (16), range 26-100 yr] in which intraoperative fluid management was guided by SPV (trigger: SPV>10%). Central venous O2 saturation (ScvO2), lactate and bilirubin, creatinine, indocyanine green plasma disappearance rate (ICG-PDR), and gastric mucosal CO(2) tension were measured after induction of anaesthesia, after 3, 6, 12, and 24 h. RESULTS: Patient characteristics, duration of surgery [5.8 (2.5) vs 5.4 (2.5) h], and infusion volumes (median 4865 vs 4330 ml) were comparable between the groups. At 3 and 6 h, SPV (P=0.04, P=0.01) and Deltadown (P=0.005, P=0.01) were significantly higher in the control group. Oxygen transport and organ function were comparable: baseline and 24 h values for ICG PDR: 28.5 (7.9) and 22.7 (7.8) vs 23.9 (6.9) and 26.1 (5.9)% min(-1), 77.7 (6.6) and 72.6 (5.5) vs 79.3 (7.1) and 72.8 (6.7)% for ScvO2 and 1.0 (0.4) and 1.2 (0.6) vs 0.9 (0.2) and 1.3 (0.5) mmol litre(-1) for lactate. Length of mechanical ventilation, ICU stay, and mortality were comparable. CONCLUSIONS: In comparison with routine care, intraoperative SPV-guided treatment was associated with slightly increased fluid adminstration whereas organ perfusion and function was similar. PMID- 18511440 TI - Psychology of pain. AB - This article briefly reviews psychological aspects of pain, paying special attention to chronic pain. The review considers the interruptive and interference effects of pain and its impact on a person's identity. The importance of processes related to interruption, interference, and identity will vary across people and the duration of pain. Although brief phasic pain such as that presented in the laboratory will have marked interruptive effects, it is unlikely to produce interference or impact on a person's identity. Acute clinical pain will have both interruptive and interference effects, albeit of a temporary nature, but it is unlikely to have any impact on a person's identity. Chronic persistent pain or frequent recurrent episodic pain, such as headache, may have profound effects on a Person's life. Each of these themes is illustrated with examples drawn from the experimental and clinical literature. PMID- 18511441 TI - Neuropathic pain: emerging treatments. AB - Neuropathic pain remains one of the most challenging of all neurological diseases and presents a large unmet need for improved therapies. Many mechanistic details are still lacking, but greater knowledge of overlapping mechanisms and disease comorbidities has highlighted key areas for intervention. These include peripheral and central hyperexcitability. Among the molecular drivers are ion channels (Nav1.7, Nav1.8, Nav1.3, Cav2.2, and alpha2-delta subunits) whose expression is changed during neuropathic pain and their block shows therapeutic utility. Block of a number of ligand-gated channels [transient receptor potential (TRP)V1, TRPM8, and neuronal nicotinic receptors (NNRs)], important in neural sensitization, may also prove beneficial. Other approaches, such as the modulation of peripheral excitability via CB1 receptors, reduction of spinal excitability through block of glutamate receptors (metabotropic glutamate receptor 5 and alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate), block of activated spinal neuroglial (CCR2 and P2X7), or increasing spinal inhibition by enhancing monoaminergic activity, all offer exciting opportunities currently being validated in the clinic. Finally of note is the emergence of biological approaches, for example, antibodies, siRNA, gene therapy, offering powerful therapeutic additions with which to redress the neurological disease imbalances causing neuropathic pain. PMID- 18511442 TI - Comment on: Thallas-Bonke et al. (2008) Inhibition of NADPH oxidase prevents advanced glycation end product-mediated damage in diabetic nephropathy through a protein kinase C-alpha-dependent pathway: Diabetes 57:460-469, 2008. PMID- 18511444 TI - From fibrosis to sclerosis: mechanisms of glomerulosclerosis in diabetic nephropathy. PMID- 18511445 TI - Oxidative stress as a major culprit in kidney disease in diabetes. AB - It is postulated that localized tissue oxidative stress is a key component in the development of diabetic nephropathy. There remains controversy, however, as to whether this is an early link between hyperglycemia and renal disease or develops as a consequence of other primary pathogenic mechanisms. In the kidney, a number of pathways that generate reactive oxygen species (ROS) such as glycolysis, specific defects in the polyol pathway, uncoupling of nitric oxide synthase, xanthine oxidase, NAD(P)H oxidase, and advanced glycation have been identified as potentially major contributors to the pathogenesis of diabetic kidney disease. In addition, a unifying hypothesis has been proposed whereby mitochondrial production of ROS in response to chronic hyperglycemia may be the key initiator for each of these pathogenic pathways. This postulate emphasizes the importance of mitochondrial dysfunction in the progression and development of diabetes complications including nephropathy. A mystery remains, however, as to why antioxidants per se have demonstrated minimal renoprotection in humans despite positive preclinical research findings. It is likely that the utility of current study approaches, such as vitamin use, may not be the ideal antioxidant strategy in human diabetic nephropathy. There is now an increasing body of data to suggest that strategies involving a more targeted antioxidant approach, using agents that penetrate specific cellular compartments, may be the elusive additive therapy required to further optimize renoprotection in diabetes. PMID- 18511446 TI - Fatty acid oxidation and insulin action: when less is more. PMID- 18511447 TI - Dedifferentiation for replication of human beta-cells: a division between mice and men? PMID- 18511448 TI - Smad1 as a biomarker for diabetic nephropathy. PMID- 18511449 TI - HNF4A and diabetes: injury before insult? PMID- 18511450 TI - Proteins in human myeloid leukemia cell line HL60 reacting with retinoic acid monoclonal antibodies. AB - The vitamin A derivative, retinoic acid (RA) has various biological effects in mammalian cells and tissues. It is well known that RA induces differentiation of leukemia cells and inhibits cell growth. There are two pathways for RA action; one via RA nuclear receptors (RARs), and one via acylation of proteins by RA (retinoylation). However, an understanding of which actions of RA occur via RARs and which occur via retinoylation is lacking. Thus, we undertook the examination of HL60 proteins using anti-RA monoclonal antibodies (ARMAs). These ARMAs showed specific binding to proteins in a saturable manner depending on protein and antibody concentration. Proteins eluted by Mono Q anion exchange chromatography and separated using two-dimensional polyacrylamide gel electrophoresis were detected by ARMAs. One of these ARMA-bound proteins in HL60 cells was identified as alpha-actinin. These results indicate that retinoylated proteins in HL60 cells can be recognized by ARMAs and that alpha-actinin modified by RA may play a significant role in RA-induced differentiation, including the promotion of cytomorphology changes. PMID- 18511451 TI - Important amino acid residues that confer CYP2C19 selective activity to CYP2C9. AB - Although CYP2C9 and CYP2C19 display 91% sequence identity at the amino acid level, the two enzymes have distinct substrate specificities for compounds such as diclofenac, progesterone and (S)-mephenytoin. Amino acid substitutions in CYP2C9 were made based on an alignment of CYP2C9, CYP2C19 and monkey CYP2C43 sequences. Mutants of CYP2C9 were expressed in Escherichia coli. Sixteen amino acids, which are common to both CYP2C19 and CYP2C43 but different between CYP2C9 and CYP2C19, were substituted in CYP2C9 (CYP2C9-16aa). Next, the mutated amino acids in CYP2C9-16aa were individually reverted to those of CYP2C9 to examine the effect of each substitution on the enzymatic activity for CYP2C marker substrates. In addition, the role of the F-G loop in CYP2C9 and CYP2C19 was examined for substrate specificity and enzymatic activity. Our results showed: (i) CYP2C9-16aa displays 11% (S)-mephenytoin 4'-hydroxylase and full omeprazole 5 hydroxylase activity compared with that of CYP2C19; (ii) residue 286 is important for conferring CYP2C9-like enzyme activity on CYP2C9-16aa and residue 442 in CYP2C19 may be involved in the interaction with NADPH-P450 reductase; (iii) substitution of the F-G loop in CYP2C9 to that of CYP2C19 enhances tolbutamide p methyhydroxylase and diclofenac 4'-hydroxylase activities and confers partial (S) mephenytoin 4'-hydroxylase and omeprazole 5-hydroxylase activities, which are attributed to CYP2C19. PMID- 18511452 TI - Crystal structure of pyruvate kinase from Geobacillus stearothermophilus. AB - The pyruvate kinase (PK) from a moderate thermophile, Geobacillus stearothermophilus, is an allosteric enzyme activated by AMP and ribose 5 phosphate but not fructose 1, 6-bisphosphate (FBP), which is a common activator of PKs. It has an extra C-terminal sequence (ECTS), which contains a highly conserved phosphoenolpyruvate (PEP) binding motif, but its function and structure remain unclear. To elucidate the structural characteristics of the effector binding site and the ECTS, the crystal structure of the C9S/C268S mutant of the enzyme was determined at 2.4 A resolution. The crystal belonged to space group P6(2)22, with unit cell parameters a, b = 145.97 A, c = 118.03 A. The enzyme was a homotetramer and its overall domain structure was similar to the previously solved structures except that the ECTS formed a new domain (C' domain). The structure of the C' domain closely resembled that of the PEP binding domain of maize pyruvate phosphate dikinase. A sulphate ion was found in a pocket in the effector-binding C domain. This site corresponds to the 6-phosphate group-binding site in yeast PK bound FBP and seems to be the effector-binding site. Through comparison of the structure of the putative effector-binding site to that of the FBP binding site of the yeast enzyme, the structural basis of the effector specificity of the G. stearothermophilus PK is discussed. PMID- 18511453 TI - Kruppel-like factor 4 is required for the expression of vascular smooth muscle cell differentiation marker genes induced by all-trans retinoic acid. AB - Kruppel-like factor 4 (KLF4) is involved in phenotypic modulation of vascular smooth muscle cells (VSMCs). All-trans retinoic acid (ATRA) inhibits VSMC proliferation and induces VSMC differentiation. However, the role of KLF4 in ATRA elicited VSMC phenotypic modulation remains unclear. Here, we show that treatment of VSMCs with ATRA resulted in significant inhibition of proliferation and migration of VSMCs, as well as up-regulation of KLF4 and the VSMC differentiation marker genes SM22alpha and SM alpha-actin (alpha-SMA). At the same time, the KLF4 target gene p53 was up-regulated, while the VSMC dedifferentiation marker gene nonmuscle myosin heavy chain-B (SMemb) was down-regulated. We also show that overexpression of KLF4 in VSMCs increased the expression of p53, SM22alpha and alpha-SMA, but decreased the expression of SMemb and VSMC proliferation and migration. Silencing of KLF4 expression by KLF4-specific small interfering RNA (siRNA) abrogated the inducing effects of ATRA on p53, SM22alpha and alpha-SMA expression and neutralized the inhibitory effects of ATRA on SMemb expression and VSMC proliferation and migration. Thus, our data suggest that KLF4 is required for the expression of VSMC differentiation marker genes induced by ATRA and that this transcription factor is one of the key mediators of retinoid actions in VSMCs. PMID- 18511454 TI - ATP-binding on fibroblast growth factor 2 partially overlaps with the heparin binding domain. AB - Fibroblast growth factor 2 (FGF2), an intensively studied heparin-binding cytokine, is an important modulator of cell growth and differentiation under both physiological and pathophysiological conditions. It has been shown recently that ATP binds to FGF2 and that this binding is crucial for its biological function. In this study we demonstrated that divalent cations were not necessary for binding of ATP to FGF2, but it could be demonstrated that heparin blocked the labelling of FGF2 with ATP indicating an involvement of the heparin-binding domain (aa 128-144) in ATP-binding. FGF2, bound to Heparin Sepharose, could be eluted with ATP and GTP, but not with cAMP, AMP or ADP. Successive mutation of positively charged amino acid residues located in the heparin-binding domain drastically reduced the signal intensity of [gamma-(32)P]ATP labelled FGF2 indicating that this domain is not only important for heparin binding to FGF2 but also for ATP-binding. PMID- 18511455 TI - Novel variants in human Aquaporin-4 reduce cellular water permeability. AB - Cerebral edema contributes significantly to morbidity and mortality after brain injury and stroke. Aquaporin-4 (AQP4), a water channel expressed in astrocytes, plays a key role in brain water homeostasis. Genetic variants in other aquaporin family members have been associated with disease phenotypes. However, in human AQP4, only one non-synonymous single-nucleotide polymorphism (nsSNP) has been reported, with no characterization of protein function or disease phenotype. We analyzed DNA from an ethnically diverse cohort of 188 individuals to identify novel AQP4 variants. AQP4 variants were constructed by site-directed mutagenesis and expressed in cells. Water permeability assays in the cells were used to measure protein function. We identified 24 variants in AQP4 including four novel nsSNPs (I128T, D184E, I205L and M224T). We did not observe the previously documented M278T in our sample. The nsSNPs found were rare ( approximately 1-2% allele frequency) and heterozygous. Computational analysis predicted reduced function mutations. Protein expression and membrane localization were similar for reference AQP4 and the five AQP4 mutants. Cellular assays confirmed that four variant AQP4 channels reduced normalized water permeability to between 26 and 48% of the reference (P < 0.001), while the M278T mutation increased normalized water permeability (P < 0.001). We identified multiple novel AQP4 SNPs and showed that four nsSNPs reduced water permeability. The previously reported M278T mutation resulted in gain of function. Our experiments provide insight into the function of the AQP4 protein. These nsSNPs may have clinical implications for patients with cerebral edema and related disorders. PMID- 18511456 TI - RNAi-mediated knockdown of dystrophin expression in adult mice does not lead to overt muscular dystrophy pathology. AB - Duchenne muscular dystrophy (DMD) is a fatal muscle wasting disorder caused by mutations in the dystrophin gene. DMD has a complex and as yet incompletely defined molecular pathophysiology. The peak of the pathology attributed to dystrophin deficiency happens between 3 and 8 weeks of age in mdx mice, the animal model of DMD. Accordingly, we hypothesized that the pathology observed with dystrophin deficiency may be developmentally regulated. Initially, we demonstrated that profound small interfering RNA-mediated dystrophin knockdown could be achieved in mouse primary muscle cultures. The use of adeno-associated virus vectors to express short-hairpin RNAs targeting dystrophin in skeletal muscle in vivo yielded a potent and specific dystrophin knockdown, but only after approximately 5 months, indicating the very long half-life of dystrophin. Interestingly, and in contrast to what is observed in congenital dystrophin deficiency, long-term ( approximately 1 year) dystrophin knockdown in adult mice did not result, per se, in overt dystrophic pathology or upregulation of utrophin. This supports our hypothesis and suggests new pathophysiology of the disease. Furthermore, taking into account the rather long half-life of dystrophin, and the notion that the development of pathology is age-dependent, it indicates that a single gene therapy approach before the onset of pathology might convey a long-term cure for DMD. PMID- 18511458 TI - RNAi-mediated knockdown of the XIP-type endoxylanase inhibitor gene, OsXIP, has no effect on grain development and germination in rice. AB - OsXIP (Oryza sativa xylanase inhibitor protein) is a XIP-type xylanase inhibitor which was identified as a protein encoded by a wound stress-responsive gene in rice. Although the OsXIP gene was specifically expressed in mature grains under basal conditions, recombinant OsXIP had no effect on rice endogenous xylanases, and OsXIP-suppressed transgenic rice plants did not exhibit any change in grain development and germination, suggesting that rice development may be independent of OsXIP. Analysis using an OsXIP-specific antibody revealed that OsXIP is markedly accumulated in apoplast in rice root cells by wounding. These results reinforced the possibility that OsXIP is involved in plant defense mechanisms against phytopathogens. PMID- 18511457 TI - Cell-lineage regulated myogenesis for dystrophin replacement: a novel therapeutic approach for treatment of muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is characterized in skeletal muscle by cycles of myofiber necrosis and regeneration leading to loss of muscle fibers and replacement with fibrotic connective and adipose tissue. The ongoing activation and recruitment of muscle satellite cells for myofiber regeneration results in loss of regenerative capacity in part due to proliferative senescence. We explored a method whereby new myoblasts could be generated in dystrophic muscles by transplantation of primary fibroblasts engineered to express a micro dystrophin/enhanced green fluorescent protein (muDys/eGFP) fusion gene together with a tamoxifen-inducible form of the myogenic regulator MyoD [MyoD-ER(T)]. Fibroblasts isolated from mdx(4cv) mice, a mouse model for DMD, were efficiently transduced with lentiviral vectors expressing muDys/eGFP and MyoD-ER(T) and underwent myogenic conversion when exposed to tamoxifen. These cells could also be induced to differentiate into muDys/eGFP-expressing myocytes and myotubes. Transplantation of transduced mdx(4cv) fibroblasts into mdx(4cv) muscles enabled tamoxifen-dependent regeneration of myofibers that express muDys. This lineage control method therefore allows replenishment of myogenic stem cells using autologous fibroblasts carrying an exogenous dystrophin gene. This strategy carries several potential advantages over conventional myoblast transplantation methods including: (i) the relative simplicity of culturing fibroblasts compared with myoblasts, (ii) a readily available cell source and ease of expansion and (iii) the ability to induce MyoD gene expression in vivo via administration of a medication. Our study provides a proof of concept for a novel gene/stem cell therapy technique and opens another potential therapeutic approach for degenerative muscle disorders. PMID- 18511459 TI - Plant inner membrane anion channel (PIMAC) function in plant mitochondria. AB - To date, the existence of the plant inner membrane anion channel (PIMAC) has been shown only in potato mitochondria, but its physiological role remains unclear. In this study, by means of swelling experiments in K(+) and ammonium salts, we characterize a PIMAC-like anion-conducting pathway in mitochondria from durum wheat (DWM), a monocotyledonous species phylogenetically far from potato. DWM were investigated since they possess a very active potassium channel (PmitoK(ATP)), so implying a very active matching anion uniport pathway and, possibly, a coordinated function. As in potato mitochondria, the electrophoretic uptake of chloride and succinate was inhibited by matrix [H(+)], propranolol, and tributyltin, and was insensitive to Mg(2+), N,N'-dicyclohexylcarbodiimide (DCCD) and mercurials, thus showing PIMAC's existence in DWM. PIMAC actively transports dicarboxylates, oxodicarboxylates, tricarboxylates and Pi. Interestingly, a novel mechanism of swelling in ammonium salts of isolated plant mitochondria is reported, based on electrophoretic anion uptake via PIMAC and ammonium uniport via PmitoK(ATP). PIMAC is inhibited by physiological compounds, such as ATP and free fatty acids, by high electrical membrane potential (Delta Psi), but not by acyl-CoAs or reactive oxygen species. PIMAC was found to cooperate with dicarboxylate carrier by allowing succinate uptake that triggers succinate/malate exchange in isolated DWM. Similar results were obtained using mitochondria from the dicotyledonous species topinambur, so suggesting generalization of results. We propose that PIMAC is normally inactive in vivo due to ATP and Delta Psi inhibition, but activation may occur in mitochondria de-energized by PmitoK(ATP) (or other dissipative systems) to replace or integrate the operation of classical anion carriers. PMID- 18511460 TI - GeConT 2: gene context analysis for orthologous proteins, conserved domains and metabolic pathways. AB - The Gene Context Tool (GeConT) allows users to visualize the genomic context of a gene or a group of genes and their orthologous relationships within fully sequenced bacterial genomes. The new version of the server incorporates information from the COG, Pfam and KEGG databases, allowing users to have an integrated graphical representation of the function of genes at multiple levels, their phylogenetic distribution and their genomic context. The sequence of any of the genes can be easily retrieved, as well as the 5' or 3' regulatory regions, greatly facilitating further types of analysis. GeConT 2 is available at: http://bioinfo.ibt.unam.mx/gecont. PMID- 18511461 TI - Development of a single-chain, quasi-dimeric zinc-finger nuclease for the selective degradation of mutated human mitochondrial DNA. AB - The selective degradation of mutated mitochondrial DNA (mtDNA) molecules is a potential strategy to re-populate cells with wild-type (wt) mtDNA molecules and thereby alleviate the defective mitochondrial function that underlies mtDNA diseases. Zinc finger nucleases (ZFNs), which are nucleases conjugated to a zinc finger peptide (ZFP) engineered to bind a specific DNA sequence, could be useful for the selective degradation of particular mtDNA sequences. Typically, pairs of complementary ZFNs are used that heterodimerize on the target DNA sequence; however, conventional ZFNs were ineffective in our system. To overcome this, we created single-chain ZFNs by conjugating two FokI nuclease domains, connected by a flexible linker, to a ZFP with an N-terminal mitochondrial targeting sequence. Here we show that these ZFNs are efficiently transported into mitochondria in cells and bind mtDNA in a sequence-specific manner discriminating between two 12 bp long sequences that differ by a single base pair. Due to their selective binding they cleave dsDNA at predicted sites adjacent to the mutation. When expressed in heteroplasmic cells containing a mixture of mutated and wt mtDNA these ZFNs selectively degrade mutated mtDNA, thereby increasing the proportion of wt mtDNA molecules in the cell. Therefore, mitochondria-targeted single-chain ZFNs are a promising candidate approach for the treatment of mtDNA diseases. PMID- 18511462 TI - Differential binding of quadruplex structures of muscle-specific genes regulatory sequences by MyoD, MRF4 and myogenin. AB - Four myogenic regulatory factors (MRFs); MyoD, Myf-5, MRF4 and Myogenin direct muscle tissue differentiation. Heterodimers of MRFs with E-proteins activate muscle-specific gene expression by binding to E-box motifs d(CANNTG) in their promoters or enhancers. We showed previously that in contrast to the favored binding of E-box by MyoD-E47 heterodimers, homodimeric MyoD associated preferentially with quadruplex structures of regulatory sequences of muscle specific genes. To inquire whether other MRFs shared the DNA binding preferences of MyoD, the DNA affinities of hetero- and homo-dimeric MyoD, MRF4 and Myogenin were compared. Similarly to MyoD, heterodimers with E47 of MRF4 or Myogenin bound E-box more tightly than quadruplex DNA. However, unlike homodimeric MyoD or MRF4, Myogenin homodimers associated weakly and nonpreferentially with quadruplex DNA. By reciprocally switching basic regions between MyoD and Myogenin we demonstrated dominance of MyoD in determining the quadruplex DNA-binding affinity. Thus, Myogenin with an implanted MyoD basic region bound quadruplex DNA nearly as tightly as MyoD. However, a grafted Myogenin basic region did not diminish the high affinity of homodimeric MyoD for quadruplex DNA. We speculate that the dissimilar interaction of MyoD and Myogenin with tetrahelical domains in muscle gene promoters may differently regulate their myogenic activities. PMID- 18511463 TI - Single-molecule imaging of full protein synthesis by immobilized ribosomes. AB - How folding of proteins is coupled to their synthesis remains poorly understood. Here, we apply single-molecule fluorescence imaging to full protein synthesis in vitro. Ribosomes were specifically immobilized onto glass surfaces and synthesis of green fluorescent protein (GFP) was achieved using modified commercial Protein Synthesis using Recombinant Elements that lacked ribosomes but contained purified factors and enzyme that are required for translation in Escherichia coli. Translation was monitored using a GFP mutant (F64L/S65T/F99S/M153T/V163A) that has a high fluorophore maturation rate and that contained the Secretion Monitor arrest sequence to prevent dissociation from the ribosome. Immobilized ribosomal subunits were labeled with Cy3 and GFP synthesis was measured by colocalization of GFP fluorescence with the ribosome position. The rate of appearance of colocalized ribosome GFP was equivalent to the rates of fluorescence appearance coupled with translation measured in bulk, and the ribosome-polypeptide complexes were stable for hours. The methods presented here are applicable to single molecule investigation of translational initiation, elongation and cotranslational folding. PMID- 18511464 TI - A RecB-family nuclease motif in the Type I restriction endonuclease EcoR124I. AB - The Type I restriction-modification enzyme EcoR124I is an ATP-dependent endonuclease that uses dsDNA translocation to locate and cleave distant non specific DNA sites. Bioinformatic analysis of the HsdR subunits of EcoR124I and related Type I enzymes showed that in addition to the principal PD-(E/D)xK Motifs, I, II and III, a QxxxY motif is also present that is characteristic of RecB-family nucleases. The QxxxY motif resides immediately C-terminal to Motif III within a region of predicted alpha-helix. Using mutagenesis, we examined the role of the Q and Y residues in DNA binding, translocation and cleavage. Roles for the QxxxY motif in coordinating the catalytic residues or in stabilizing the nuclease domain on the DNA are discussed. PMID- 18511465 TI - Transcriptional regulation of the Drosophila moira and osa genes by the DREF pathway. AB - The DNA replication-related element binding factor (DREF) plays an important role in regulation of cell proliferation in Drosophila, binding to DRE and activating transcription of genes carrying this element in their promoter regions. Overexpression of DREF in eye imaginal discs induces a rough eye phenotype in adults, which can be suppressed by half dose reduction of the osa or moira (mor) genes encoding subunits of the BRM complex. This ATP-dependent chromatin remodeling complex is known to control gene expression and the cell cycle. In the 5' flanking regions of the osa and mor genes, DRE and DRE-like sequences exist which contribute to their promoter activities. Expression levels and promoter activities of osa and mor are decreased in DREF knockdown cells and our results in vitro and in cultured cells indicate that transcription of osa and mor is regulated by the DRE/DREF regulatory pathway. In addition, mRNA levels of other BRM complex subunits and a target gene, string/cdc25, were found to be decreased by knockdown of DREF. These results indicate that DREF is involved in regulation of the BRM complex and thereby the cell cycle. PMID- 18511466 TI - Using inferred residue contacts to distinguish between correct and incorrect protein models. AB - MOTIVATION: The de novo prediction of 3D protein structure is enjoying a period of dramatic improvements. Often, a remaining difficulty is to select the model closest to the true structure from a group of low-energy candidates. To what extent can inter-residue contact predictions from multiple sequence alignments, information which is orthogonal to that used in most structure prediction algorithms, be used to identify those models most similar to the native protein structure? RESULTS: We present a Bayesian inference procedure to identify residue pairs that are spatially proximal in a protein structure. The method takes as input a multiple sequence alignment, and outputs an accurate posterior probability of proximity for each residue pair. We exploit a recent metagenomic sequencing project to create large, diverse and informative multiple sequence alignments for a test set of 1656 known protein structures. The method infers spatially proximal residue pairs in this test set with good accuracy: top-ranked predictions achieve an average accuracy of 38% (for an average 21-fold improvement over random predictions) in cross-validation tests. Notably, the accuracy of predicted 3D models generated by a range of structure prediction algorithms strongly correlates with how well the models satisfy probable residue contacts inferred via our method. This correlation allows for confident rejection of incorrect structural models. AVAILABILITY: An implementation of the method is freely available at http://www.doe-mbi.ucla.edu/services. PMID- 18511467 TI - Efficient functional clustering of protein sequences using the Dirichlet process. AB - MOTIVATION: Automatic clustering of protein sequences is an important problem in computational biology. The recent explosion in genome sequences has given biological researchers a vast number of novel protein sequences. However, the majority of these sequences have no experimental evidence for their molecular function in the cell, and the responsibility for correctly annotating these sequences falls upon the bioinformatics community. Ideally, we would like to be able to group sequences of similar or identical molecular function in an automatic fashion, without relying on experimental evidence. RESULTS: In this article I present a novel probabilistic framework that models subfamilies within a known protein family. Given a multiple sequence alignment, the model uses Dirichlet mixture densities to estimate amino acid preferences within subfamily clusters, and places a Dirichlet process prior on the overall set of clusters. Based on results from several datasets, the model breaks data accurately into functional subgroups. AVAILABILITY: The algorithm is implemented as c++ software available at bpg-research.berkeley.edu/approximately duncanb/dpcluster/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 18511468 TI - Ontologizer 2.0--a multifunctional tool for GO term enrichment analysis and data exploration. AB - The Ontologizer is a Java application that can be used to perform statistical analysis for overrepresentation of Gene Ontology (GO) terms in sets of genes or proteins derived from an experiment. The Ontologizer implements the standard approach to statistical analysis based on the one-sided Fisher's exact test, the novel parent-child method, as well as topology-based algorithms. A number of multiple-testing correction procedures are provided. The Ontologizer allows users to visualize data as a graph including all significantly overrepresented GO terms and to explore the data by linking GO terms to all genes/proteins annotated to the term and by linking individual terms to child terms. AVAILABILITY: The Ontologizer application is available under the terms of the GNU GPL. It can be started as a WebStart application from the project homepage, where source code is also provided: http://compbio.charite.de/ontologizer. REQUIREMENTS: Ontologizer requires a Java SE 5.0 compliant Java runtime engine and GraphViz for the optional graph visualization tool. PMID- 18511469 TI - CellTrack: an open-source software for cell tracking and motility analysis. AB - MOTIVATION: Cell motility is a critical part of many important biological processes. Automated and sensitive cell tracking is essential to cell motility studies where the tracking results can be used for diagnostic or curative decisions and where mathematical models can be developed to deepen our understanding of the mechanisms underlying cell motility. RESULTS: We have developed CellTrack: a self-contained, extensible, and cross-platform software package for cell tracking and motility analysis. Besides the general purpose image enhancement, object segmentation and tracking algorithms, we have implemented a novel edge-based method for sensitive tracking of the cell boundaries, and constructed an ensemble of methods that achieves refined tracking results even under large displacements or deformations of the cells. AVAILABILITY: CellTrack is an Open Source project and is freely available at http://db.cse.ohio-state.edu/CellTrack. PMID- 18511470 TI - Exploring the use of recombinant human TSH in the diagnosis of central hypothyroidism. AB - CONTEXT: The diagnosis of central hypothyroidism (CH) is often difficult to establish as serum TSH levels may be low, normal, or slightly increased. OBJECTIVE: To explore the use of recombinant human TSH (rhTSH) in the diagnosis of CH. DESIGN: Randomized single-blind clinical trial. SETTING: Outpatient clinic of a tertiary care referral center. INTERVENTION: A single intramuscular injection of 0.1 and 0.9 mg rhTSH in random order with 1-week interval. PARTICIPANTS: Eighteen adult patients with pituitary insufficiency and six healthy age-, sex-, and body mass index-matched controls. Six patients had untreated CH (newCH), six had treated CH (CH), and six patients were TSH sufficient (nonCH). Five weeks before TSH stimulation, levothyroxine was replaced with tri-iodothyronine (T(3)) for 4 weeks. One week before stimulation, treatment was withdrawn. MAIN OUTCOME MEASURES: Thyroid hormones and thyroglobulin (Tg) before and 2, 3(1/2), 7, 24, 48, and 72 h after each injection. RESULTS: In the newCH group, basal free thyroxine (FT(4)) levels were lower than in controls (P<0.05). After 0.9 mg rhTSH, the increases in FT(4) and reverse T(3) (rT(3)) were less marked in the newCH group than in controls (FT(4)+/-s.e.m. 9.2+/-0.5 to 19.7+/-1.2 vs 11.3+/-0.5 to 27.8.2+/-2.4 pmol/l, P<0.05). The CH group exhibited reduced basal and stimulated FT(4) compared with the TSH-sufficient groups. Tg increased similarly among all study groups after rhTSH injection. CONCLUSION: In this pilot study, patients with untreated CH had lower response to 0.9 mg rhTSH in FT(4) and rT(3) than controls. An rhTSH test may be useful in the diagnosis of CH, but further studies are required. PMID- 18511471 TI - Subclinical thyroid dysfunction and mortality: an estimate of relative and absolute excess all-cause mortality based on time-to-event data from cohort studies. AB - OBJECTIVES: To what extent persons with subclinical hyper- or hypothyroidism are more (or less) likely to die than euthyroid control subjects remains a matter of controversy. METHODS: We searched electronic reference databases up to July 31, 2007. Three reviewers independently assessed eligibility. Cohort studies published in full that reported data on the hazard ratio (HR) for mortality from all causes in persons with subclinical thyroid dysfunction versus euthyroid controls were included. RESULTS: Based on seven cohorts including 290 participants with subclinical hyperthyroidism, random-effects models estimated that the pooled HR for all-cause mortality was 1.41 (95% confidence interval (CI), 1.12-1.79; P=0.004). Using the pooled HR and standard life-table methods applied to a US reference population, we estimated that a white US woman, when diagnosed with subclinical hyperthyroidism at age of 70, has an excess mortality of 1.5, 4.0, and 8.7% at 2, 5, and 10 years respectively after diagnosis. Likewise, a white US man has an excess mortality of 2.3, 5.7, and 10.7%. For the nine cohorts including 1580 participants with subclinical hypothyroidism, observed heterogeneity (Q test P=0.006; I(2)=63%) disappeared after pooling cohorts in predefined subgroups according to the presence or absence of a comorbid condition. In doing so, the pooled HR for all-cause mortality was 1.03 (95% CI, 0.78-1.35; P=0.83) in cohorts from the community and 1.76 (95% CI, 1.36 2.30; P<0.001) in cohorts of participants with comorbidities (P=0.014 for heterogeneity among study groups). CONCLUSIONS: Individuals with subclinical hyperthyroidism demonstrate a 41% increase in relative mortality from all causes versus euthyroid control subjects. Mathematical modeling suggests that absolute excess mortality after diagnosis might depend on age, with an increase beyond the age of 60, especially in aging men. For patients with subclinical hypothyroidism, the relative risk of all-cause mortality is increased only in patients with comorbid conditions. PMID- 18511472 TI - Incretin levels in polycystic ovary syndrome. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) has been linked to a high risk of type 2 diabetes mellitus. Disturbances in the secretion of the incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) have been observed in states with impaired glucose regulation. This paper considers the secretion of GIP and GLP-1 after oral glucose load in a group of lean, glucose-tolerant PCOS women in comparison with age- and body mass index (BMI)-matched healthy women. DESIGN: Case control. METHODS: PCOS (n=21, 25.8+/ 4.1 years, BMI 21.6+/-1.7 kg/m(2)) and control healthy women (CT, n=13, 28.5+/ 7.2 years, BMI 20.3+/-2.5 kg/m(2)) underwent oral glucose tolerance test (OGTT) with blood sampling for glucose, insulin, C-peptide, total GIP, and active GLP-1. Insulin sensitivity was determined both at fasting and during the test. STATISTICS: Repeated measures ANOVA. RESULTS: Glucose levels and insulin sensitivity did not differ between PCOS and CT. PCOS had significantly higher levels of C-peptide (P<0.05) and tended to have higher insulin levels. The levels of total GIP were significantly higher in PCOS than in CT (P<0.001). Active GLP-1 levels exhibited a significantly different time-dependent pattern in PCOS (P<0.002 for PCOS versus time interaction). GLP-1 concentrations were similar in PCOS and CT in the early phase of OGTT and then reached significantly lower levels in PCOS than in CT at 180 min (P<0.05). CONCLUSIONS: Increased total GIP and lower late phase active GLP-1 concentrations during OGTT characterize PCOS women with higher C-peptide secretion in comparison with healthy controls, and may be the early markers of a pre-diabetic state. PMID- 18511473 TI - Resistin in serum is associated with higher levels of IL-1Ra in post-menopausal women with rheumatoid arthritis. AB - OBJECTIVES: The aim of this study was to investigate associations between serum levels of resistin, an adipokine and markers of inflammation, bone metabolism, plasma lipids and kidney function in post-menopausal RA patients and to evaluate if HRT during 2 yrs affected resistin levels. METHODS: Eighty-eight women were randomly allocated to receive HRT, vitamin D(3) and calcium or vitamin D(3) and calcium alone. Serum levels of resistin, IL-1beta, IL-1 receptor antagonist (IL 1Ra), IL-6, IL-6 soluble receptor, TNF-alpha were measured by ELISA, markers of bone metabolism, carboxyterminal cross-linked telopeptide of type I collagen (ICTP) and carboxyterminal propeptide of type I procollagen by RIA, ESR, CRP, Hb, creatinine and lipids by standard laboratory techniques, BMD and total lean mass (TLM) by DXA and joint destruction by Larsen score. Resistin was also measured in 42 healthy control women. RESULTS: There was no difference in resistin concentration between patients and healthy controls. Resistin was significantly correlated with IL-1Ra, CRP, TNF-alpha, ICTP, glucocorticosteroids and Larsen score and inversely with BMD, hip and with TLM. In multiple regression analysis, IL-1Ra, TLM and use of corticosteroids remained determinants of resistin. Patients treated with HRT displayed significant increase in resistin compared with controls in the first but not the second year. CONCLUSIONS: Resistin was associated with increased inflammation, particularly by the acute-phase reactant IL-1Ra antagonizing IL-1beta, joint destruction, glucocorticosteroids and with reduced BMD and TLM. These findings suggest resistin being a significant mediator in the inflammatory process in RA. Further studies examining the mechanisms behind the relation between resistin and IL-1Ra are encouraged. HRT does not seem to have important long-term effect on resistin. PMID- 18511474 TI - Angiotensin-converting enzyme inhibitors delay the occurrence of renal involvement and are associated with a decreased risk of disease activity in patients with systemic lupus erythematosus--results from LUMINA (LIX): a multiethnic US cohort. AB - OBJECTIVE: To examine if angiotensin-converting enzyme (ACE) inhibitor use delays the occurrence of renal involvement and decreases the risk of disease activity in SLE patients. METHODS: SLE patients (Hispanics, African Americans and Caucasians) from the lupus in minorities: nature vs nurture (LUMINA) cohort were studied. Renal involvement was defined as ACR criterion and/or biopsy-proven lupus nephritis. Time-to-renal involvement was examined by univariable and multivariable Cox proportional hazards regression analyses. Disease activity was examined with a case-crossover design and a conditional logistic regression model; in the case intervals, a decrease in the SLAM-R score >or=4 points occurred but not in the control intervals. RESULTS: Eighty of 378 patients (21%) were ACE inhibitor users; 298 (79%) were not. The probability of renal involvement free-survival at 10 yrs was 88.1% for users and 75.4% for non-users (P = 0.0099, log rank test). Users developed persistent proteinuria and/or biopsy proven lupus nephritis (7.1%) less frequently than non-users (22.9%), P = 0.016. By multivariable Cox proportional hazards regression analyses, ACE inhibitors use [hazard ratio (HR) 0.27; 95% CI 0.09, 0.78] was associated with a longer time-to renal involvement occurrence whereas African American ethnicity (HR 3.31; 95% CI 1.44, 7.61) was with a shorter time. ACE inhibitor use (54/288 case and 254/1148 control intervals) was also associated with a decreased risk of disease activity (HR 0.56; 95% CI 0.34, 0.94). CONCLUSIONS: ACE inhibitor use delays the development of renal involvement and associates with a decreased risk of disease activity in SLE; corroboration of these findings in other lupus cohorts is desirable before practice recommendations are formulated. PMID- 18511475 TI - The incidence of rheumatoid arthritis in Spain: results from a nationwide primary care registry. AB - OBJECTIVE: To estimate the incidence of early arthritis (EA) and of RA in adults (>16 yrs) in Spain. METHODS: Primary care physicians were instructed in the detection of new cases using a checklist. All cases were evaluated at EA units (EAUs) within 15 days of detection. ACR criteria for the classification of RA were assessed every 6 months thereafter. RESULTS: In an area covering 4,342,378 inhabitants over the age of 16 yrs, 2467 patients were referred to the EAU, of whom 1063 fulfilled EA criteria (43.1%). After 6 months, 362 patients fulfilled RA criteria. The estimated annual incidence of EA was 25/100,000 population (95% CI: 23, 26). The annual incidence of RA was 8.3 cases/100,000 (95% CI: 7.5, 9.2): 11.3/100,000 in women (95% CI: 10.0, 12.8), and 5.2/100,000 in men (95% CI: 4.3, 6.3). The incidence of RA increased with age in both sexes. At the 6 months' assessment, 187 (51.7%) of the patients with RA were RF positive. The presentation of RA was mainly polyarticular (n = 268; 74.0%). There were 701/1063 patients with EA who did not fulfil RA criteria by 6 months after the first rheumatologist visit. If all cases of undifferentiated arthritis (n = 118; 17%) became RA, the incidence would be in the range of 10 cases/100,000 population. CONCLUSIONS: RA incidence in Spain is in the lower range of published data. The incidence of EA is about three times that of RA. PMID- 18511476 TI - Cost-effectiveness of advising the use of topical or oral ibuprofen for knee pain; the TOIB study [ISRCTN: 79353052]. AB - OBJECTIVE: Advice to use topical or oral NSAIDs is equally effective for the treatment of knee pain in older people. The ingredient cost of topical preparations is typically more than oral preparations, but could save costs because they have fewer adverse effects. A cost-utility study is needed to decide on their comparative cost effectiveness. METHODS: We recruited 585 people aged >or=50 yrs with knee pain; 282 participated in a randomized controlled trial and 303 in a patient preference study from 26 MRC General Practice Research Framework practices in the UK. They received advice to preferentially use topical or oral NSAIDs for knee pain. We calculated the comparative cost per quality-adjusted life year (QALY) from both a National Health Service (NHS) and a societal perspective over 12 and 24 months. RESULTS: Compared with the topical route, oral NSAIDs cost the NHS pound191 and pound72 more over 1 yr in the randomized trial and preference study, respectively. The cost per QALY, from an NHS perspective, was in the range of pound9000- pound12000 in the randomized trial. In the preference study, it was pound2564 over 1 yr, but over 2 yrs the oral route was dominant. CONCLUSIONS: Our cost-effectiveness analysis supports the use of oral NSAIDs in selected patients. Nevertheless, deciding to recommend oral NSAIDs in preference to topical NSAIDs could have a substantial impact on NHS costs because of the uncertainty in the cost-effectiveness estimate. PMID- 18511477 TI - Rapid identification of infants for antiretroviral therapy in a resource poor setting: the Kenya experience. AB - In Kenya, HIV diagnosis is not routinely carried out in infants, and yet rapid diagnosis could improve access to lifesaving interventions. A cheap and readily accessible service can resolve this problem, if feasible. In this pilot study the feasibility and costs of provision of an infant HIV diagnosis service in Kenya are evaluated. Dried blood spots (DBS) were collected from infants exposed to HIV, sent to a central testing laboratory and tested using the Roche Amplicor v. 1.5 DNA PCR kit. The results were then dispatched to health facilities within a week. A total of 15.4% of the samples tested HIV+ despite the widespread access to prevention of mother to child transmission (PMTCT) programs in Kenya. The cost per test at 21.50 USD is prohibitive and will limit access to diagnosis. It remains to be seen whether the increase in testing will immediately lead to an increase in access to antiretroviral therapy (ART) services for infants. PMID- 18511478 TI - Behavioral analyses of sugar processing in choice, feeding, and learning in larval Drosophila. AB - Gustatory stimuli have at least 2 kinds of function: They can support immediate, reflexive responses (such as substrate choice and feeding) and they can drive internal reinforcement. We provide behavioral analyses of these functions with respect to sweet taste in larval Drosophila. The idea is to use the dose-effect characteristics as behavioral "fingerprints" to dissociate reflexive and reinforcing functions. For glucose and trehalose, we uncover relatively weak preference. In contrast, for fructose and sucrose, preference responses are strong and the effects on feeding pronounced. Specifically, larvae are attracted to, and feeding is stimulated most strongly for, intermediate concentrations of either sugar: Using very high concentrations (4 M) results in weakened preference and suppression of feeding. In contrast to such an optimum function regarding choice and feeding, an asymptotic dose-effect function is found for reinforcement learning: Learning scores reach asymptote at 2 M and remain stable for a 4-M concentration. A similar parametric discrepancy between the reflexive (choice and feeding) and reinforcing function is also seen for sodium chloride (Niewalda T, Singhal S, Fiala A, Saumweber T, Wegener S, Gerber B, in preparation). We discuss whether these discrepancies are based either on inhibition from high-osmolarity sensors upon specifically the reflexive pathways or whether different sensory pathways, with different effective dose-response characteristics, may have preferential access to drive either reflex responses or modulatory neurons mediating internal reinforcement, respectively. PMID- 18511479 TI - Chronic forearm pain presents as a transient and indistinct pain site in a community setting: results from a UK population survey. AB - OBJECTIVES: To investigate the prevalence of chronic forearm pain in a non occupational community setting over a 2-year period. METHOD: A longitudinal community-based postal questionnaire survey conducted in the south-eastern quadrant of England. RESULTS: We received 2493/4172 (60%) responses at baseline and we followed up 429 of these 2 years later: 252 responded (59%). Forearm pain prevalence was 4% at baseline and 5% at follow-up. Over 95% of those with forearm pain had pain in other areas [odds ratio 1.5 (95% confidence interval 1.3-1.7)] and it was most commonly associated with elbow and wrist pain. Seventy-six per cent of those with forearm pain at baseline recovered. At follow-up, 78% of those with chronic forearm pain had new-onset forearm pain. CONCLUSIONS: Persistent forearm pain (pain for over 2 years) was rare and the capacity for recovery was good (76%). Isolated forearm pain as a diagnostic category is of little utility. Treating and managing forearm pain in a site-specific manner is unlikely to be successful owing to its strong association with pain in other areas. In the community, forearm pain laterality was not evident; our findings suggest that forearm pain in the workplace is influenced by different factors to those in a community setting. PMID- 18511480 TI - Robust and sensitive iTRAQ quantification on an LTQ Orbitrap mass spectrometer. AB - Isobaric stable isotope tagging reagents such as tandem mass tags or isobaric tags for relative and absolute quantification enable multiplexed quantification of peptides via reporter ion signals in the low mass range of tandem mass spectra. Until recently, the poor recovery of low mass fragments observed in tandem mass spectra acquired on ion trap mass spectrometers precluded the use of these reagents on this widely available instrument platform. The Pulsed Q Dissociation (PQD) technique allows negotiating this limitation but suffers from poor fragmentation efficiency, which has raised doubts in the community as to its practical utility. Here we show that by carefully optimizing instrument parameters such as collision energy, activation Q, delay time, ion isolation width, number of microscans, and number of trapped ions, low m/z fragment ion intensities can be generated that enable accurate peptide quantification at the 100 amol level. Side by side comparison of PQD on an LTQ Orbitrap with CID on a five-year old Q-Tof Ultima using complex protein digests shows that whereas precision of quantification of 10-15% can be achieved by both approaches, PQD quantifies twice as many proteins. PQD on an LTQ Orbitrap also outperforms "higher energy collision induced dissociation" on the same instrument using the recently introduced octapole collision cell in terms of lower limit of quantification. Finally, we demonstrate the significant analytical potential of iTRAQ quantification using PQD on an LTQ Orbitrap by quantitatively measuring the kinase interaction profile of the small molecule drug imatinib in K-562 cells. This article gives practical guidance for the implementation of PQD, discusses its merits, and for the first time, compares its performance to higher energy collision-induced dissociation. PMID- 18511481 TI - An integrated, directed mass spectrometric approach for in-depth characterization of complex peptide mixtures. AB - LC-MS/MS has emerged as the method of choice for the identification and quantification of protein sample mixtures. For very complex samples such as complete proteomes, the most commonly used LC-MS/MS method, data-dependent acquisition (DDA) precursor selection, is of limited utility. The limited scan speed of current mass spectrometers along with the highly redundant selection of the most intense precursor ions generates a bias in the pool of identified proteins toward those of higher abundance. A directed LC-MS/MS approach that alleviates the limitations of DDA precursor ion selection by decoupling peak detection and sequencing of selected precursor ions is presented. In the first stage of the strategy, all detectable peptide ion signals are extracted from high resolution LC-MS feature maps or aligned sets of feature maps. The selected features or a subset thereof are subsequently sequenced in sequential, non redundant directed LC-MS/MS experiments, and the MS/MS data are mapped back to the original LC-MS feature map in a fully automated manner. The strategy, implemented on an LTQ-FT MS platform, allowed the specific sequencing of 2,000 features per analysis and enabled the identification of more than 1,600 phosphorylation sites using a single reversed phase separation dimension without the need for time-consuming prefractionation steps. Compared with conventional DDA LC-MS/MS experiments, a substantially higher number of peptides could be identified from a sample, and this increase was more pronounced for low intensity precursor ions. PMID- 18511482 TI - GABAergic signalling to adult-generated neurons. AB - New neurons are continuously generated in discrete regions of the adult brain. In the hippocampus, newly generated cells undergo a step-wise progression of maturation that is regulated at multiple stages by a variety of physiological and pathological stimuli. Neural progenitors and newborn neurons initially receive exclusively GABAergic synaptic input, and accumulating evidence suggests that depolarizing actions of GABA contribute to activity-dependent regulation. Here we provide a brief overview of GABAergic signalling to newborn neurons in the hippocampus and describe how it regulates adult neurogenesis. PMID- 18511483 TI - Novel aspects of the molecular mechanisms controlling insulin secretion. AB - Pancreatic beta-cells secrete insulin by Ca(2+)-dependent exocytosis of secretory granules. beta-cell exocytosis involves SNARE (soluble NSF-attachment protein receptor) proteins similar to those controlling neurotransmitter release and depends on the close association of L-type Ca(2+) channels and granules. In most cases, the secretory granules fuse individually but there is ultrastructural and biophysical evidence of multivesicular exocytosis. Estimates of the secretory rate in beta-cells in intact islets indicate a release rate of approximately 15 granules per beta-cell per second, 100-fold higher than that observed in biochemical assays. Single-vesicle capacitance measurements reveal that the diameter of the fusion pore connecting the granule lumen with the exterior is approximately 1.4 nm. This is considerably smaller than the size of insulin and membrane fusion is therefore not obligatorily associated with release of the cargo, a feature that may contribute to the different rates of secretion detected by the biochemical and biophysical measurements. However, small molecules like ATP and GABA, which are stored together with insulin in the granules, are small enough to be released via the narrow fusion pore, which accordingly functions as a molecular sieve. We finally consider the possibility that defective fusion pore expansion accounts for the decrease in insulin secretion observed in pathophysiological states including long-term exposure to lipids. PMID- 18511484 TI - Initial segment Kv2.2 channels mediate a slow delayed rectifier and maintain high frequency action potential firing in medial nucleus of the trapezoid body neurons. AB - The medial nucleus of the trapezoid body (MNTB) is specialized for high frequency firing by expression of Kv3 channels, which minimize action potential (AP) duration, and Kv1 channels, which suppress multiple AP firing, during each calyceal giant EPSC. However, the outward K(+) current in MNTB neurons is dominated by another unidentified delayed rectifier. It has slow kinetics and a peak conductance of approximately 37 nS; it is half-activated at -9.2 +/- 2.1 mV and half-inactivated at -35.9 +/- 1.5 mV. It is blocked by several non-specific potassium channel antagonists including quinine (100 microm) and high concentrations of extracellular tetraethylammonium (TEA; IC(50) = 11.8 mM), but no specific antagonists were found. These characteristics are similar to recombinant Kv2-mediated currents. Quantitative RT-PCR showed that Kv2.2 mRNA was much more prevalent than Kv2.1 in the MNTB. A Kv2.2 antibody showed specific staining and Western blots confirmed that it recognized a protein approximately 110 kDa which was absent in brainstem tissue from a Kv2.2 knockout mouse. Confocal imaging showed that Kv2.2 was highly expressed in axon initial segments of MNTB neurons. In the absence of a specific antagonist, Hodgkin-Huxley modelling of voltage-gated conductances showed that Kv2.2 has a minor role during single APs (due to its slow activation) but assists recovery of voltage-gated sodium channels (Nav) from inactivation by hyperpolarizing interspike potentials during repetitive AP firing. Current-clamp recordings during high frequency firing and characterization of Nav inactivation confirmed this hypothesis. We conclude that Kv2.2-containing channels have a distinctive initial segment location and crucial function in maintaining AP amplitude by regulating the interspike potential during high frequency firing. PMID- 18511485 TI - Calprotectin is released from human skeletal muscle tissue during exercise. AB - Skeletal muscle has been identified as a secretory organ. We hypothesized that IL 6, a cytokine secreted from skeletal muscle during exercise, could induce production of other secreted factors in skeletal muscle. IL-6 was infused for 3 h into healthy young males (n = 7) and muscle biopsies obtained at time points 0, 3 and 6 h in these individuals and in resting controls. Affymetrix microarray analysis of gene expression changes in skeletal muscle biopsies identified a small set of genes changed by IL-6 infusion. RT-PCR validation confirmed that S100A8 and S100A9 mRNA were up-regulated 3-fold in skeletal muscle following IL-6 infusion compared to controls. Furthermore, S100A8 and S100A9 mRNA levels were up regulated 5-fold in human skeletal muscle following cycle ergometer exercise for 3 h at approximately 60% of in young healthy males (n = 8). S100A8 and S100A9 form calprotectin, which is known as an acute phase reactant. Plasma calprotectin increased 5-fold following acute cycle ergometer exercise in humans, but not following IL-6 infusion. To identify the source of calprotectin, healthy males (n = 7) performed two-legged dynamic knee extensor exercise for 3 h with a work load of approximately 50% of peak power output and arterial-femoral venous differences were obtained. Arterial plasma concentrations for calprotectin increased 2-fold compared to rest and there was a net release of calprotectin from the working muscle. In conclusion, IL-6 infusion and muscle contractions induce expression of S100A8 and S100A9 in skeletal muscle. However, IL-6 alone is not a sufficient stimulus to facilitate release of calprotectin from skeletal muscle. PMID- 18511486 TI - The untiring search for the most complete proteome representation: reviewing the methods. AB - Proteomic research has proved valuable for understanding the molecular mechanisms of biological processes, as well as in the search for biomarkers for a variety of diseases which lack a molecular diagnostic. While several new approaches are being developed, two-dimensional (2-DE) gel electrophoresis is still one of the most commonly used techniques, despite its many limitations. However, for biomarker research, 2-DE gel electrophoresis alone does not fulfill the necessary pre-requisites. If such a technique is utilized exclusively, a great part of a given proteome remains unseen. Therefore, very precise and sensitive techniques are needed. Here, we present a brief review of known methodologies that try to overcome the limitations of conventional proteome analysis as well as their respective advantages and limitations. PMID- 18511487 TI - Permanent embryo arrest: molecular and cellular concepts. AB - Developmental arrest is one of the mechanisms responsible for the elevated levels of embryo demise during the first week of in vitro development. Approximately 10 15% of IVF embryos permanently arrest in mitosis at the 2- to 4-cell cleavage stage showing no indication of apoptosis. Reactive oxygen species (ROS) are implicated in this process and must be controlled in order to optimize embryo production. A stress sensor that can provide a key understanding of permanent cell cycle arrest and link ROS with cellular signaling pathway(s) is p66Shc, an adaptor protein for apoptotic-response to oxidative stress. Deletion of the p66Shc gene in mice results in extended lifespan, which is linked to their enhanced resistance to oxidative stress and reduced levels of apoptosis. p66Shc has been shown to generate mitochondrial H(2)O(2) to trigger apoptosis, but may also serve as an integration point for many signaling pathways that affect mitochondrial function. We have detected elevated levels of p66Shc and ROS within arrested embryos and believe that p66Shc plays a central role in regulating permanent embryo arrest. In this paper, we review the cellular and molecular aspects of permanent embryo arrest and speculate on the mechanism(s) and etiology of this method of embryo demise. PMID- 18511488 TI - Health insurance and cervical cancer screening among older women in Latin American and Caribbean cities. AB - BACKGROUND: The purpose of this study is to describe the prevalence of Papanicolaou (Pap) smear use for cervical cancer screening and to estimate its association with type of health care insurance. METHODS: A cross-sectional study using data from the Health, Well-Being and Aging in Latin America and the Caribbean Study (SABE). The sample includes 6357 women aged 60 and older from seven cities. The outcome was reporting a Pap smear for cervical cancer screening during the previous 2 years. Main independent variable was health care insurance. Covariates were demographic or socioeconomic variables, medical conditions and functional status. RESULTS: Prevalence of Pap smear use across the seven cities ranged from 21% in Bridgetown to 45% in Mexico City. In a multivariate analysis of the combined sample, without Havana that has universal health care insurance, women with public insurance (OR 0.55, 95% CI 0.43-0.71) or with no insurance (OR 0.23, 95% CI 0.15-0.34) were less likely to have a Pap smear compared with women with private insurance. Also, women with no insurance were less likely to have a Pap smear (OR 0.40, 95% CI 0.30-0.54) compared with women with any health insurance. CONCLUSIONS: In general, the prevalence of Pap smear use was lower than that reported for Hispanic populations in the United States. Overall, lack of health insurance or having public health insurance determined lower odds for having a Pap smear for cervical cancer screening. PMID- 18511489 TI - Vulnerability to heat-related mortality in Latin America: a case-crossover study in Sao Paulo, Brazil, Santiago, Chile and Mexico City, Mexico. AB - BACKGROUND: Factors affecting vulnerability to heat-related mortality are not well understood. Identifying susceptible populations is of particular importance given anticipated rising temperatures from climatic change. METHODS: We investigated heat-related mortality for three Latin American cities (Mexico City, Mexico; Sao Paulo, Brazil; Santiago, Chile) using a case-crossover approach for 754 291 deaths from 1998 to 2002. We considered lagged exposures, confounding by air pollution, cause of death and susceptibilities by educational attainment, age and sex. RESULTS: Same and previous day apparent temperature were most strongly associated with mortality risk. Effect estimates remained positive though lowered after adjustment for ozone or PM(10). Susceptibility increased with age in all cities. The increase in mortality risk for those >or=65 comparing the 95th and 75th percentiles of same-day apparent temperature was 2.69% (95% CI: -2.06 to 7.88%) for Santiago, 6.51% (95% CI: 3.57-9.52%) for Sao Paulo and 3.22% (95% CI: 0.93-5.57%) for Mexico City. Patterns of vulnerability by education and sex differed across communities. Effect estimates were higher for women than men in Mexico City, and higher for men elsewhere, although results by sex were not appreciably different for any city. In Sao Paulo, those with less education were more susceptible, whereas no distinct patterns by education were observed in the other cities. CONCLUSIONS: Elevated temperatures are associated with mortality risk in these Latin American cities, with the strongest associations in Sao Paulo, the hottest city. The elderly are an important population for targeted prevention measures, but vulnerability by sex and education differed by city. PMID- 18511490 TI - Exposure to ultraviolet radiation and risk of malignant lymphoma and multiple myeloma--a multicentre European case-control study. AB - BACKGROUND: Three recent studies have reported a decreased risk of non-Hodgkin lymphoma (NHL) for high ultraviolet (UV) radiation exposure. METHODS: We conducted a multicentre case-control study during 1998-2004 in France, Germany, Ireland, Italy and Spain, comprising 1518 cases of NHL, 268 cases of Hodgkin lymphoma, 242 cases of multiple myeloma and 2124 population or hospital controls. We collected information on sensitivity to sun and personal exposure to UV radiation in childhood and adulthood via interview, and assessed occupational exposure to UV radiation from the occupational history. RESULTS: The risk of Hodgkin and NHL was increased for increasing skin sensitivity to the sun [odds ratio (OR) for no suntan vs very brown 2.35, 95% CI 0.94-5.87 and 1.39, 95% CI 1.03-1.87, respectively]. The risk of diffuse large B-cell lymphoma was reduced for increasing adult personal (OR for highest vs lowest quartile of exposure in free days 0.62, 95% CI 0.44-0.87) and for occupational exposure to UV radiation (OR for highest vs lowest exposure tertile 0.63, 95% CI 0.37-1.04). The risk of multiple myeloma was increased for personal exposure to UV radiation during adulthood (OR for highest vs lowest quartile of exposure in free days 1.49, 95% CI 0.88-2.50). A protective effect was observed for use of sun lamps for diffuse large B-cell lymphoma (OR for 25+ times vs never 0.63, 95% CI 0.38-1.03). CONCLUSIONS: The hypothesis of a protective effect of UV radiation on lymphoma is supported by our results. The underlying mechanisms might differ from those operating in skin carcinogenesis. The increased risk of multiple myeloma is worth replication. PMID- 18511491 TI - Gender and effects of a common genetic variant in the NOS1 regulator NOS1AP on cardiac repolarization in 3761 individuals from two independent populations. AB - BACKGROUND: A longer heart-rate corrected QT interval (QTc) is associated with increased risk of ventricular arrhythmias. Women have longer resting QTc and are more likely than men to develop drug-induced QT prolongation. Recent studies have shown association between resting QTc and a common variant (rs10494366) of the NOS1 regulator, NOS1AP. We investigated the association between rs10494366 in NOS1AP and QTc, and assessed gender-specific NOS1AP associations with QTc during rest and after exercise. METHODS: We investigated the SNP associations with resting QTc in 919 women and 918 men from 504 representative families in the UK GRAPHIC study, and with QTc at rest and at 3 min recovery after exercise in 699 women and 1225 men referred for exercise testing in the Finnish FINCAVAS study. RESULTS: In the GRAPHIC study the minor allele (G) of the NOS1AP SNP rs10494366 prolonged QTc by 4.59 ms (95% CI 2.77-6.40; P = 7.63/10(7)) in women, but only by 1.62 ms (95% CI -0.15 to 3.38; P = 0.073) in men (gender-SNP interaction term P = 0.025). In the FINCAVAS study the G allele significantly prolonged QTc in both women (P = 0.0063) and men (P = 0.0043) at 3 min recovery after exercise, but at rest an association was only seen in women (P = 0.020 excluding outliers). CONCLUSIONS: A common NOS1AP variant prolongs QTc with a difference between genders. Further studies should aim to confirm this finding and to assess whether NOS1AP genotype influences the risk of drug-induced QT prolongation and risk of consequent arrhythmias. PMID- 18511492 TI - Demographic, health services and socio-economic factors associated with pulmonary tuberculosis mortality in Los Altos Region of Chiapas, Mexico. AB - BACKGROUND: Chiapas is one of the Mexican states having the highest rates of Pulmonary Tuberculosis (PTB), due to the numerous factors impeding its management and control (poverty, poor housing and nutrition, shortage of health resources, among others). OBJECTIVES: To analyse the PTB mortality of a cohort of patients in Los Altos Region of Chiapas, who had been diagnosed with PTB from January 1, 1998 to December 31, 2002; and, to identify demographic, socioeconomic and health services utilization factors, associated with death from PTB. METHODS: Analysis of a cohort of patients aged over 14 years diagnosed with PTB in the above mentioned period (n = 431) in Los Altos region of Chiapas. The records of the Tuberculosis Programme were reviewed, and patients were located through a search attempting to locate them in their homes. Those found alive were interviewed and asked to provide sputum samples. In the case of deceased patients, a verbal autopsy was obtained from a member of their family. RESULTS: The records of the PTB Programme in the area were incomplete and erroneous in many cases. The results of the home follow-up visits were: 208 (48%) patients located alive, five of whom were still PTB positive (three with multi-drug resistance); 145 (34%) could not be located and 78 (18%) had already died. Apparently, in at least 40 cases, the deaths were associated with PTB. Of these forty, 33 (83%) died without having received any medical care. The factors associated with dying from PTB were: 45 and over years of age (OR = 1.3; 95% CI = 0.98-1.3), 0-3 schooling years (OR = 3.3; 95% CI = 1.1-9.6), engaged in agriculture (OR = 2.2; 95% CI = 1.1 4.4), not living in main villages of their municipality (OR = 1.2; 95% CI = 1.0 1.3), living in a rural community (OR = 2.7; 95% CI = 1.1-6.8), not having been treated in DOTS (OR = 1.2; 95% CI = 1.0-1.3) and having defaulted from treatment (OR = 11.5; 95% CI = 5.3-24.8). CONCLUSIONS: The high rate of mortality due to PTB observed constitutes a serious public health problem deserving attention. There is an urgent need to evaluate and restructure the Tuberculosis Programme in the studied area. PMID- 18511493 TI - Cultural differences in musculoskeletal symptoms and disability. AB - OBJECTIVES: To test the hypothesis that cultural factors such as health beliefs and expectations have an important influence on common musculoskeletal symptoms and associated disability, we compared prevalence rates in groups of workers carrying out similar physical activities in different cultural settings. METHODS: We conducted a cross-sectional survey at factories and offices in Mumbai, India and in the UK. A questionnaire about symptoms, disability and risk factors was administered at interview to six occupational groups: three groups of office workers who regularly used computer keyboards (165 Indian, 67 UK of Indian subcontinental origin and 172 UK white), and three groups of workers carrying out repetitive manual tasks with the hands or arms (178 Indian, 73 UK of Indian subcontinental origin and 159 UK white). Modified Cox regression was used to calculate hazard ratios (HRs) for the prevalence of symptoms and disability by occupational group, adjusted for differences in sex, age, mental health and job satisfaction. RESULTS: Reported occupational activities were similar in the three groups of office workers (frequent use of keyboards) and in the three groups of manual workers (frequent movements of the wrist or fingers, bending of the elbow, work with the hands above shoulder height and work with the neck twisted). In comparison with the Indian manual workers, the prevalence of back, neck and arm pain was substantially higher in all of the other five occupational groups. The difference was greatest for arm pain lasting >30 days in the past year in UK white manual workers (HR 17.8, 95% CI 5.4-59.1) and UK manual workers of Indian subcontinental origin (HR 20.5, 95% CI 5.7-73.1). Office workers in India had lower rates of pain in the wrist and hand than office workers in the UK. Only 1% of the Indian manual workers and 16% of the Indian office workers had ever heard of 'RSI' or similar terms, as compared with 80% of the UK workers. CONCLUSIONS: Our findings support the hypothesized impact of cultural factors on common musculoskeletal complaints. Current controls on hazardous physical activities in the workplace may not have the benefits that would be predicted from observational epidemiology. PMID- 18511494 TI - Steroidogenic factor 1 regulates expression of the cannabinoid receptor 1 in the ventromedial hypothalamic nucleus. AB - The nuclear receptor steroidogenic factor 1 (SF-1) plays essential roles in the development and function of the ventromedial hypothalamic nucleus (VMH). Considerable evidence links the VMH and SF-1 with the regulation of energy homeostasis. Here, we demonstrate that SF-1 colocalizes in VMH neurons with the cannabinoid receptor 1 (CB1R) and that a specific CB1R agonist modulates electrical activity of SF-1 neurons in hypothalamic slice preparations. We further show that SF-1 directly regulates CB1R gene expression via a SF-1 responsive element at -101 in its 5'-flanking region. Finally, we show that knockout mice with selective inactivation of SF-1 in the brain have decreased expression of CB1R in the region of the VMH and exhibit a blunted response to systemically administered CB1R agonists. These studies suggest that SF-1 directly regulates the expression of CB1R, which has been implicated in the regulation of energy homeostasis and anxiety-like behavior. PMID- 18511495 TI - The Xenopus laevis isoform of G protein-coupled receptor 3 (GPR3) is a constitutively active cell surface receptor that participates in maintaining meiotic arrest in X. laevis oocytes. AB - Oocytes are held in meiotic arrest in prophase I until ovulation, when gonadotropins trigger a subpopulation of oocytes to resume meiosis in a process termed "maturation." Meiotic arrest is maintained through a mechanism whereby constitutive cAMP production exceeds phosphodiesterase-mediated degradation, leading to elevated intracellular cAMP. Studies have implicated a constitutively activated Galpha(s)-coupled receptor, G protein-coupled receptor 3 (GPR3), as one of the molecules responsible for maintaining meiotic arrest in mouse oocytes. Here we characterized the signaling and functional properties of GPR3 using the more amenable model system of Xenopus laevis oocytes. We cloned the X. laevis isoform of GPR3 (XGPR3) from oocytes and showed that overexpressed XGPR3 elevated intraoocyte cAMP, in large part via Gbetagamma signaling. Overexpressed XGPR3 suppressed steroid-triggered kinase activation and maturation of isolated oocytes, as well as gonadotropin-induced maturation of follicle-enclosed oocytes. In contrast, depletion of XGPR3 using antisense oligodeoxynucleotides reduced intracellular cAMP levels and enhanced steroid- and gonadotropin-mediated oocyte maturation. Interestingly, collagenase treatment of Xenopus oocytes cleaved and inactivated cell surface XGPR3, which enhanced steroid-triggered oocyte maturation and activation of MAPK. In addition, human chorionic gonadotropin treatment of follicle-enclosed oocytes triggered metalloproteinase-mediated cleavage of XGPR3 at the oocyte cell surface. Together, these results suggest that GPR3 moderates the oocyte response to maturation-promoting signals, and that gonadotropin-mediated activation of metalloproteinases may play a partial role in sensitizing oocytes for maturation by inactivating constitutive GPR3 signaling. PMID- 18511496 TI - Selective regulation of H1 histamine receptor signaling by G protein-coupled receptor kinase 2 in uterine smooth muscle cells. AB - Histamine stimulates uterine contraction; however, little is known regarding the mechanism or regulation of uterine histamine receptor signaling. Here we investigated the regulation of Galpha(q/11)-coupled histamine receptor signaling in human myometrial smooth muscle cells using the inositol 1,4,5-trisphosphate biosensor pleckstrin homology domain of phospholipase-delta1 tagged to enhanced green fluorescent protein and the Ca(2+)-sensitive dye Fluo-4. Histamine addition caused concentration-dependent increases in inositol 1,4,5-trisphosphate and [Ca(2+)](i) in the ULTR human uterine smooth muscle cell line and primary human myometrial cells. These effects were completely inhibited by the H(1) histamine receptor antagonist, diphenhydramine, and were unaffected by the H(2) histamine receptor antagonist, cimetidine. ULTR and primary myometrial cells were transfected with either dominant-negative G protein-coupled receptor kinases (GRKs) or small interfering RNAs targeting specific GRKs to assess the roles of this protein kinase family in H(1) histamine receptor desensitization. Dominant negative GRK2, but not GRK5 or GRK6, prevented H(1) histamine receptor desensitization. Similarly, transfection with short interfering RNAs (that each caused >70% depletion of the targeted GRK) for GRK2, but not GRK3 or GRK6, also prevented H(1) histamine receptor desensitization. Our data suggest that histamine stimulates phospholipase C-signaling in myometrial smooth muscle cells through H(1) histamine receptors and that GRK2 recruitment is a key mechanism in the regulation of H(1) histamine receptor signaling in human uterine smooth muscle. These data provide insights into the in situ regulation of this receptor subtype and may inform pathophysiological functioning in preterm labor and other conditions involving uterine smooth muscle dysregulation. PMID- 18511497 TI - The nuclear receptor Rev-erbalpha is a liver X receptor (LXR) target gene driving a negative feedback loop on select LXR-induced pathways in human macrophages. AB - A role of the nuclear receptor Rev-erbalpha in the regulation of transcription pathways involving other nuclear receptors is emerging. Indeed, Rev-erbalpha is a negative regulator of transcription by binding to overlapping response elements shared with various nuclear receptors, including the peroxisome proliferator activated receptors and the retinoid-related orphan receptor alpha (RORalpha). Here, we show that Rev-erbalpha is expressed in primary human macrophages and that its expression is induced by synthetic ligands for the liver X receptors (LXRs), which control cholesterol homeostasis, inflammation, and the immune response in macrophages. LXRalpha binds to a specific response element in the human Rev-erbalpha promoter, thus inducing Rev-erbalpha transcriptional expression. Interestingly, Rev-erbalpha does not influence basal or LXR-regulated cholesterol homeostasis. However, Rev-erbalpha overexpression represses the induction of toll-like receptor (TLR)-4 by LXR agonists, whereas Rev-erbalpha silencing by short interfering RNA results in enhanced TLR-4 expression upon LXR activation. Electrophoretic mobility shift, chromatin immunoprecipitation, and transient transfection experiments demonstrate that Rev-erbalpha represses human TLR-4 promoter activity by binding as a monomer to a RevRE site overlapping with the LXR response element site in the TLR-4 promoter. These data identify Rev erbalpha as a new LXR target gene, inhibiting LXR-induction of TLR-4 in a negative transcriptional feedback loop, but not cholesterol homeostasis gene expression. PMID- 18511498 TI - Aquaporin 1 is important for maintaining secretory granule biogenesis in endocrine cells. AB - Aquaporins (AQPs), a family of water channels expressed in epithelial cells, function to transport water in a bidirectional manner to facilitate transepithelial fluid absorption and secretion. Additionally, AQP1 and AQP5 are found in pancreatic zymogen granules and synaptic vesicles and are involved in vesicle swelling and exocytosis in exocrine cells and neurons. Here, we show AQP1 is in dense-core secretory granule (DCSG) membranes of endocrine tissue: pituitary and adrenal medulla. The need for AQP1 in endocrine cell function was examined by stable transfection of AQP1 antisense RNA into AtT20 cells, a pituitary cell line, to down-regulate AQP1 expression. These AQP1-deficient cells showed more than 60% depletion of DCSGs and significantly decreased DCSG protein levels, including proopiomelanocotin/pro-ATCH and prohormone convertase 1/3, but not non-DCSG proteins. Pulse-chase studies revealed that whereas DCSG protein synthesis was unaffected, approximately 50% of the newly synthesized proopiomelanocortin was degraded within 1 h. Low levels of ACTH were released upon stimulation, indicating that the small number of DCSGs that were made in the presence of the residual AQP1 were functionally competent for exocytosis. Analysis of anterior pituitaries from AQP1 knockout mice showed reduced prohormone convertase 1/3, carboxypeptidase E, and ACTH levels compared to wild type mice demonstrating that our results observed in AtT20 cells can be extended to the animal model. Thus, AQP1 is important for maintaining DCSG biogenesis and normal levels of hormone secretion in pituitary endocrine cells. PMID- 18511499 TI - Endocrine-related resources from the National Institutes of Health. PMID- 18511500 TI - A computational screen for mouse signaling pathways targeted by microRNA clusters. AB - MicroRNAs (miRNAs) are one class of short, endogenous RNAs which can regulate gene expression at the post-transcriptional level. Previous analysis revealed that mammalian miRNAs tend to cluster on chromosomes. However, the functional consequences of this clustering and conservation property are largely unknown. In this study we present a method to identify signaling pathways targeted by clustered miRNAs. We performed a computational screen for mouse signaling pathways targeted by miRNA clusters. Here, we report that the target genes of 3 miRNA clusters are overrepresented in 15 signaling pathways. We provided experimental evidence that one miRNA cluster, mmu-mir-183-96-182 targets Irs1, Rasa1, and Grb2, all of which are located in the insulin signaling pathway. Theses results suggest that by targeting components with different roles along a signaling pathway, different members of one miRNA cluster can act as a whole to coordinately control the signal transduction process. PMID- 18511501 TI - Distinctive structures between chimpanzee and human in a brain noncoding RNA. AB - Human accelerated region 1 (HAR1) is a short DNA region identified recently to have evolved the most rapidly among highly constrained regions since the divergence from our common ancestor with chimpanzee. It is transcribed as part of a noncoding RNA specifically expressed in the developing human neocortex. Employing a panoply of enzymatic and chemical probes, our analysis of HAR1 RNA proposed a secondary structure model differing from that published. Most surprisingly, we discovered that the substitutions between the chimpanzee and human sequences led the human HAR1 RNA to adopt a cloverleaf-like structure instead of an extended and unstable hairpin in the chimpanzee sequence. Thus, the rapid evolutionary changes resulted in a profound rearrangement of HAR1 RNA structure. Altogether, our results provide a structural context for elucidating HAR1 RNA function. PMID- 18511502 TI - Isoform-specific increases in murine skeletal muscle peroxisome proliferator activated receptor-gamma coactivator-1alpha (PGC-1alpha) mRNA in response to beta2-adrenergic receptor activation and exercise. AB - Adrenergic receptor (AR) activation increases expression of peroxisome proliferator-activated receptor (PPAR)-gamma coactivator 1alpha (PGC-1alpha) mRNA, which may promote mitochondrial biogenesis in skeletal muscles. An AR activated increase in PGC-1alpha mRNA was observed in exercise. PGC-1alpha mRNA is considered a single transcript (PGC-1alpha-a); however, a transcript search of PGC-1alpha in expressed sequence tag libraries revealed that two novel isoforms of PGC-1alpha mRNA, named PGC-1alpha-b and PGC-1alpha-c, were expressed in mice tissues. Compared with PGC-1alpha-a mRNA (a previously described isoform), PGC 1alpha-b or PGC-1alpha-c mRNA was transcribed by a different exon 1 of the PGC 1alpha gene and produced slightly smaller-sized proteins. PGC-1alpha-b or PGC 1alpha-c protein was functional; both isoforms possessed transcriptional activity and could coactivate PPARs, similar to those in PGC-1alpha-a in vitro. Transgenic mice overexpressing PGC-1alpha-b or PGC-1alpha-c in skeletal muscles showed increased gene expression related to mitochondrial biogenesis and fatty acid oxidation. In C57BL/6J mice, injection of the beta2-AR agonist clenbuterol increased PGC-1alpha-b and PGC-1alpha-c mRNA expression more than 350-fold, but not PGC-1alpha-a, in skeletal muscle. A single bout of exercise also increased PGC-1alpha-b and PGC-1alpha-c mRNAs, but not PGC-1alpha-a, in skeletal muscles. The increases in skeletal muscles in response to exercise were inhibited by pretreatment with the beta2-AR-specific inhibitor ICI 118,551. However, in liver, fasting increased PGC-1alpha-a mRNA, but not PGC-1alpha-b and PGC-1alpha-c mRNAs. These data indicate that AR activation is a major mechanism of an increase in PGC 1alpha expression in skeletal muscles, and the increase in PGC-1alpha mRNAs was isoform specific. PMID- 18511503 TI - The androgen and progesterone receptors regulate distinct gene networks and cellular functions in decidualizing endometrium. AB - Progesterone is indispensable for differentiation of human endometrial stromal cells (HESCs) into decidual cells, a process that critically controls embryo implantation. We now show an important role for androgen receptor (AR) signaling in this differentiation process. Decreased posttranslational modification of the AR by small ubiquitin-like modifier (SUMO)-1 in decidualizing cells accounted for increased responsiveness to androgen. By combining small interfering RNA technology with genome-wide expression profiling, we found that AR and progesterone receptor (PR) regulate the expression of distinct decidual gene networks. Ingenuity pathway analysis implicated a preponderance of AR-induced genes in cytoskeletal organization and cell motility, whereas analysis of AR repressed genes suggested involvement in cell cycle regulation. Functionally, AR depletion prevented differentiation-dependent stress fiber formation and promoted motility and proliferation of decidualizing cells. In comparison, PR depletion perturbed the expression of many more genes, underscoring the importance of this nuclear receptor in diverse cellular functions. However, several PR-dependent genes encode for signaling intermediates, and knockdown of PR, but not AR, compromised activation of WNT/beta-catenin, TGFbeta/SMAD, and signal transducer and activator of transcription (STAT) pathways in decidualizing cells. Thus, the nonredundant function of the AR in decidualizing HESCs, centered on cytoskeletal organization and cell cycle regulation, implies an important role for androgens in modulating fetal-maternal interactions. Moreover, we show that PR regulates HESC differentiation, at least in part, by reprogramming growth factor and cytokine signal transduction. PMID- 18511504 TI - Isolation, cloning, and expression mapping of a gene encoding an antidiuretic hormone and other CAPA-related peptides in the disease vector, Rhodnius prolixus. AB - After a blood meal, Rhodnius prolixus undergoes a rapid diuresis to eliminate excess water and salts. During the voiding of this primary urine, R. prolixus acts as a vector of Chagas' disease, with the causative agent, Trypanosoma cruzi, infecting the human host via the urine. Diuresis in R. prolixus is under the neurohormonal control of serotonin and peptidergic diuretic hormones, and thus, diuretic hormones play an important role in the transmission of Chagas' disease. Although diuretic hormones may be degraded or excreted, resulting in the termination of diuresis, it would also seem appropriate, given the high rates of secretion, that a potent antidiuretic factor could be present and act to prevent excessive loss of water and salts after the postgorging diuresis. Despite the medical importance of R. prolixus, no genes for any neuropeptides have been cloned, including obviously, those that control diuresis. Here, using molecular biology in combination with matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry, we determined the sequence of the CAPA gene and CAPA-related peptides in R. prolixus, which includes a peptide with anti-diuretic activity. We have characterized the expression of mRNA encoding these peptides in various developmental stage and also examined the tissue-specific distribution in fifth-instars. The expression is localized to numerous bilaterally paired cell bodies within the central nervous system. In addition, our results show that RhoprCAPA gene expression is also associated with the testes, suggesting a novel role for this family of peptides in reproduction. PMID- 18511505 TI - Regulation of key antioxidant enzymatic systems in the sheep endometrium by ovarian steroids. AB - Reactive oxygen species (ROS) and their control by antioxidant enzymes are involved in the physiology of the female reproductive system. Thus, it is important to understand the regulation of key antioxidant enzymatic pathways. The roles of estrogen and progesterone in regulating the physiological functions of the endometrium have become central dogma. We examined the effects of ovarian steroids on superoxide dismutases (SOD1 and SOD2), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GSR) activities in the aglandular caruncular and glandular inter-caruncular endometrial tissues of ovariectomized (OVX) ewes and in OVX ewes treated with estradiol (E2), progesterone (P4), or both hormones according to schedules designed to produce physiological changes of these hormones during the estrous cycle. The activities SOD2, CAT, GPX and GSR in both endometrial tissues were unaffected by P4 treatment. The activity of SOD1 in the aglandular tissue was unaffected by P4 treatment, however this treatment decreased SOD1 activity in the glandular tissue (P < 0.01). Treatment with E2, either alone or in combination with P4, decreased SOD1 (P < 0.01), CAT (P < 0.01) and GPX (P < 0.05) activities in both endometrial tissues. The activity of GSR decreased only in the glandular tissue (P < 0.05) after E2 treatment, either alone or in combination with P4. No change in SOD2 activity was detected in both endometrial tissues after administration of E2, P4 or both hormones. This study provides the first firm evidence for the role of ovarian steroid hormones in the regulation of the activities of key antioxidant enzyme in the endometrium of female mammals. PMID- 18511506 TI - Functional significance of a truncated thyroid receptor subtype lacking a hormone binding domain in goldfish. AB - Thyroid hormones are important mediators of growth and development in vertebrates and act by binding to a specific family of thyroid receptors (TRs). The TRs belong to the nuclear receptor superfamily, with two conserved regions, a DNA binding domain and a ligand binding domain (LBD). We recently demonstrated the presence of four TR subtypes in goldfish, two with complete DNA binding domains and LBDs (TRalpha-1 and TRbeta) and two novel forms including a transcript resembling TRalpha with variation in the LBD as well as a TRalpha-truncated (TRalpha-t) form lacking a LBD. To study the functional significance of TR subtypes, we first investigated the regulation of hepatic goldfish deiodinase type 3 (D3) by T3 and validated a bioassay in which D3 gene expression is up regulated significantly in vivo and in vitro. Using short interfering RNA, TRalpha-1, TRbeta, or TRalpha-t was specifically knocked down and thyroid hormone induced D3 gene expression was measured. Short interfering RNA against TRalpha-1 or TRbeta reduced the T3 induction of deiodinase gene expression to 50% or less than 25% of control (T3 treated) cells, respectively. Knocking down TRalpha-t alone, however, increased D3 expression 500-fold supporting the hypothesis that TRalpha-t plays a modulatory role in thyroid hormone-induced gene expression. Our results provide important insight into thyroid receptor biology in goldfish and a framework for the better understanding of thyroid receptor function in all vertebrates. PMID- 18511507 TI - Crucial role of estrogen receptor-alpha interaction with transcription coregulators in follicle-stimulating hormone and transforming growth factor beta1 up-regulation of steroidogenesis in rat ovarian granulosa cells. AB - This study was to explore estrogen receptor (ER) involvement in FSH and TGFbeta1 stimulated steroidogenesis in rat ovarian granulosa cells. We first determined the specific involvement of ERalpha and ERbeta in the process, and then investigated the molecular interaction of ERalpha and transcription coregulators in FSH and TGFbeta1 up-regulation of steroidogenic gene expression. Primary culture of ovarian granulosa cells from antral follicles of gonadotropin-primed immature rats was used. Interestingly, a selective ERalpha antagonist methyl piperidino-pyrazole (MPP) [like ER antagonist ICI-182,780 (ICI)] decreased FSH +/ TGFbeta1-stimulated progesterone production, whereas an androgen receptor antagonist hydroxyflutamide and particularly a selective ERbeta antagonist 4-[2 Phenyl-5,7-bis(trifluoromethyl) pyrazolo [1,5-a] pyrimidin-3-yl] phenol had no significant effect. Consistent with this, a selective ERbeta agonist diarylpropionitrile (unlike 17beta-estradiol) also had no effect on FSH +/- TGFbeta1-stimulated progesterone production. Furthermore, a selective ERalpha agonist 4,4',4''-(4-Propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (like 17beta estradiol) enhanced FSH-stimulated progesterone production, and this was abolished by pretreatment with MPP. Immunoblotting and chromatin immunoprecipitation analyses indicate that MPP/ICI suppression of FSH +/- TGFbeta1 action is partly attributed to the reduced ERalpha-mediated expression of Hsd3b and Cyp11a1 genes, but not steroidogenic acute regulatory protein. Furthermore, FSH +/- TGFbeta1 increased ERalpha association with histone acetylases (CBP and SRC-1) and coactivator of peroxisome proliferator-activated receptor gamma (PGC-1alpha), and MPP/ICI dramatically reduced these interactions. In addition, FSH +/- TGFbeta1 increased CBP, SRC-1, and PGC-1alpha binding to Hsd3b and Cyp11a1 genes. Together, we demonstrate for the first time that ERalpha interaction with transcription coregulators, histone acetylases (CBP/SRC-1), and PGC-1alpha is crucial to FSH and TGFbeta1-up-regulated expression of Hsd3b and Cyp11a1, and, thus, progesterone production in rat ovarian granulosa cells. PMID- 18511508 TI - Thymulin inhibits monocrotaline-induced pulmonary hypertension modulating interleukin-6 expression and suppressing p38 pathway. AB - The pathogenesis of pulmonary hypertension (PH) includes an inflammatory response. Thymulin, a zinc-dependent thymic hormone, has important immunobiological effects by inhibiting various proinflammatory cytokines and chemokines. We investigated morphological and hemodynamic effects of thymulin administration in a rat model of monocrotaline (MCT)-induced PH, as well as the pattern of proinflammatory cytokine gene expression and the intracellular pathways involved. Adult Wistar rats received an injection of MCT (60 mg/kg, sc) or an equal volume of saline. One day after, the animals randomly received during 3 wk an injection of saline, vehicle (zinc plus carboxymethyl cellulose), or thymulin (100 ng/kg, sc, daily). At d 23-25, the animals were anesthetized for hemodynamic recordings, whereas heart and lungs were collected for morphometric and molecular analysis. Thymulin prevented morphological, hemodynamic, and inflammatory cardiopulmonary profile characteristic of MCT-induced PH, whereas part of these effects were also observed in MCT-treated animals injected with the thymulin's vehicle containing zinc. The pulmonary thymulin effect was likely mediated through suppression of p38 pathway. PMID- 18511510 TI - Berberine acutely inhibits insulin secretion from beta-cells through 3',5'-cyclic adenosine 5'-monophosphate signaling pathway. AB - Berberine, a hypoglycemic agent, has recently been shown to activate AMP activated protein kinase (AMPK) contributing to its beneficial metabolic effects in peripheral tissues. However, whether berberine exerts a regulatory effect on beta-cells via AMPK or other signaling pathways and counteracts glucolipotoxicity remains uncertain. In the present study, the impact of berberine on beta-cell function was investigated in vivo and in vitro. In high-fat-fed rats, berberine treatment for 6 wk significantly decreased plasma glucose and insulin levels before and after an oral glucose challenge along with the reduction of body weight and improvement of blood lipid profile. In accordance with the in vivo results, berberine acutely decreased glucose-stimulated insulin secretion (GSIS) and palmitate-potentiated insulin secretion in MIN6 cells and rat islets. However, pretreated with berberine for 24 h augmented the response of MIN6 cells and rat islets to glucose and attenuated the glucolipotoxicity. Berberine acutely increased AMPK activity in MIN6 cells. However, compound C, an AMPK inhibitor, completely reversed troglitazone-suppressed GSIS, not berberine-suppressed GSIS. Otherwise, berberine decreased cAMP-raising agent-potentiated insulin secretion in MIN6 cells and rat islets. These results suggest that the activation of AMPK is required for troglitazone-suppressed GSIS, whereas cAMP signaling pathway contributes, at least in part, to the regulatory effect of berberine on insulin secretion. PMID- 18511511 TI - Structural interactions between kisspeptin and GnRH neurons in the mediobasal hypothalamus of the male rhesus monkey (Macaca mulatta) as revealed by double immunofluorescence and confocal microscopy. AB - Kisspeptin is recognized to play a critical role in eliciting the pubertal resurgence of pulsatile GnRH release, the proximal trigger of puberty in higher primates. Expression of the kisspeptin receptor (GPR54) by GnRH neurons indicates a direct action of kisspeptin on the GnRH neuronal network. The purpose of the present study was to examine the distribution of kisspeptin cell bodies in the monkey hypothalamus and to assess the structural basis for the stimulatory action of kisspeptin on the GnRH neuronal network. Three castrated male rhesus monkeys, 39-51 months of age, were deeply anesthetized and their brains perfused transcardially with 4% paraformaldehyde in PBS. Serial 25-microm coronal sections throughout the hypothalamus were prepared, and immunopositive neurons identified using a cocktail of specific primary antibodies (sheep anti-kisspeptin at 1:120,000, and rabbit anti-GnRH at 1:100,000) detected with fluorescently tagged secondary antibodies (antisheep, Alexa Fluor 488; antirabbit, Cy3) in combination with confocal microscopy. Kisspeptin perikarya were found only in the mediobasal hypothalamus (MBH) almost exclusively in the posterior two-thirds of the arcuate nucleus. Surprisingly, kisspeptin-beaded axons made only infrequent contacts with GnRH neurons (kisspeptin and GnRH profiles abutting in a 0.5- to 1.0-mum optical section) in the MBH. In the median eminence, kisspeptin and GnRH axons were found in extensive and intimate association. GnRH contacts on kisspeptin perikarya and dendrites were observed. These findings indicate that nonsynaptic pathways of communication in the median eminence should be considered as a possible mechanism of kisspeptin regulation of GnRH release, and provide an anatomical basis for reciprocal control of kisspeptin neuronal activity by GnRH. PMID- 18511512 TI - Tuberoinfundibular peptide of 39 residues: a neuromodulator starting a second career in the control of meiosis. PMID- 18511513 TI - Environmental estrogens and endocrine disruption: importance of comparative endocrinology. PMID- 18511514 TI - Estrogen receptor (ER) beta modulates ERalpha responses to estrogens in the developing rat ventromedial nucleus of the hypothalamus. AB - The mechanisms by which estradiol exerts specific actions on neural function are unclear. In brain the actions of estrogen receptor (ER) alpha are well documented, whereas the functions of ERbeta are not yet fully elucidated. Here, we report that ERbeta inhibits the activity of ERalpha in an anatomically specific manner within the neonatal (postnatal d 7) brain. Using selective agonists we demonstrate that the selective activation of ERalpha in the relative absence of ERbeta activation induces progesterone receptor expression to a greater extent than estradiol alone in the ventromedial nucleus, but not the medial preoptic nucleus, despite high ERalpha expression. Selective activation of ERbeta attenuates the ERalpha-mediated increase in progesterone receptor expression in the ventromedial nucleus but has no effect in medial preoptic nucleus. These results suggest that ERalpha/ERbeta interactions may regulate the effects of estrogens on neural development and reveal the neonatal brain as a unique model in which to study the specificity of steroid-induced gene expression. PMID- 18511515 TI - Liraglutide, a long-acting human glucagon-like peptide 1 analog, improves glucose homeostasis in marginal mass islet transplantation in mice. AB - The current scarcity of high-quality deceased pancreas donors prevents widespread application of islet transplantation for treatment of labile type 1 diabetes mellitus. Opportunities for the improvement of current techniques include optimization of islet isolation and purification, use of culture with pharmacological insulinotropic agents, strategies to reduce graft rejection and inflammation, and the search for alternative insulin producing tissue. Here, we report our findings on the efficacy of the long-acting human glucagon-like peptide 1 analog, liraglutide, in a mouse model of marginal mass islet transplantation. Liraglutide was administered (200 microg/kg sc twice daily) after a marginal mass syngeneic islet transplant in streptozotocin-induced diabetic BALB/c mice. Time-to-normoglycemia was significantly shorter in liraglutide-treated animals (median 1 vs. 7 d; P = 0.0003), even in recipients receiving sirolimus (median 1 vs. 72.5 d; P < 0.0001). Liraglutide-treated animals also demonstrated improved glucose tolerance as assessed by an ip glucose tolerance test. Liraglutide discontinuation at postoperative d 90 resulted in diminished glucose tolerance during the ip glucose tolerance test, whereas a late start liraglutide therapy 90 d after transplant resulted in no improvement. These findings suggest that liraglutide therapy mediates early and late insulinotropic effects. In accord with this hypothesis, insulin/terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling fluorescence microscopy showed reduced transplanted beta-cell apoptosis in liraglutide-treated recipients 48 h after transplant. In addition, liraglutide resulted in improved glucose-dependent insulin secretion. Overall, our data show that liraglutide has a beneficial impact on the engraftment and function of syngeneic islet transplants in mice, when administered continuously starting on the day of transplant. PMID- 18511516 TI - Suppression by CD4+CD25+ regulatory T cells is dependent on expression of heme oxygenase-1 in antigen-presenting cells. AB - Heme oxygenase-1 (HO-1) has been viewed as a cytoprotective protein, ameliorating the effects of inflammatory cellular damage, and as beneficial in allograft protection from acute and chronic rejection, suggesting important functions in both innate and adaptive immune responses. Mice deficient in HO-1 exhibit defective immune regulation characterized by a proinflammatory phenotype. We examined if impaired regulatory T cell (Treg) function contributes to the immunoregulatory defects observed in HO-1(-/-) mice. HO-1(-/-) mice exhibited a significantly higher proportion of Foxp3-expressing cells among total CD4(+) and CD4(+)CD25(+) cells in comparison to HO-1(+/+) mice, and HO-1(-/-) Treg cells were at least as effective as HO-1(+/+) Treg cells in suppressing proliferation of effector T cells in vitro from either HO-1(+/+) or HO-1(-/-) mice. However, the absence of HO-1 in antigen-presenting cells abolished the suppressive activity of Treg cells on effector T cells. These findings demonstrate that HO-1 activity in antigen-presenting cells is important for Treg-mediated suppression, providing an explanation for the apparent defect in immune regulation in HO-1(-/ ) mice. PMID- 18511517 TI - Cleavage of type II collagen by cathepsin K in human osteoarthritic cartilage. AB - Cathepsin K is a cysteine protease of the papain family that cleaves triple helical type II collagen, the major structural component of the extracellular matrix of articular cartilage. In osteoarthritis (OA), the anabolic/catabolic balance of articular cartilage is disrupted with the excessive cleavage of collagen II by collagenases or matrix metalloproteinases. A polyclonal antibody against a C-terminal neoepitope (C2K) generated in triple-helical type II collagen by the proteolytic action of cathepsin K was prepared and used to develop an enzyme-linked immunosorbent assay to study the generation of this epitope and the effects of its presence in normal adult and osteoarthritic femoral condylar articular cartilage. The generation of the C2K epitope in explant culture and the effect of a specific cathepsin K inhibitor were studied. The neoepitope, which is not generated by the collagenase matrix metalloproteinase-13, increased with age in articular cartilage and was significantly elevated in osteoarthritic cartilage compared with adult nonarthritic cartilage. Moreover, in explants from three of eight OA patients, the generation of the neoepitope in culture was significantly reduced by a specific, nontoxic inhibitor of cathepsin K. These data suggest that cathepsin K is involved in the cleavage of type II collagen in human articular cartilage in certain OA patients and that it may play a role in both OA pathophysiology and the aging process. PMID- 18511518 TI - The tuberous sclerosis complex regulates trafficking of glucose transporters and glucose uptake. AB - Human cancers often display an avidity for glucose, a feature that is exploited in clinical staging and response monitoring by using (18)F-fluoro-deoxyglucose (FDG) positron emission tomography. Determinants of FDG accumulation include tumor blood flow, glucose transport, and glycolytic rate, but the underlying molecular mechanisms are incompletely understood. The phosphoinositide-3 kinase/Akt/mammalian target of rapamycin complex (mTORC) 1 pathway has been implicated in this process via the hypoxia-inducible factor alpha-dependent expression of vascular endothelial growth factor and glycolytic enzymes. Thus, we predicted that tumors with elevated mTORC1 activity would be accompanied by high FDG uptake. We tested this hypothesis in eight renal angiomyolipomas in which the loss of tuberous sclerosis complex (TSC) 1/2 function gave rise to constitutive mTORC1 activation. Surprisingly, these tumors displayed low FDG uptake on positron emission tomography. Exploring the underlying mechanisms in vitro revealed that Tsc2 regulates the membrane localization of the glucose transporter proteins (Glut)1, Glut2, and Glut4, and, therefore, glucose uptake. Down regulation of cytoplasmic linker protein 170, an mTOR effector, rescued Glut4 trafficking in Tsc2(-/-) cells, whereas up-regulation of Akt activity in these cells was insufficient to redistribute Glut4 to the plasma membrane. The effect of mTORC1 on glucose uptake was confirmed using a liver-specific Tsc1- deletion mouse model in which FDG uptake was reduced in the livers of mutant mice compared with wild-type controls. Together, these data show that mTORC1 activity is insufficient for increased glycolysis in tumors and that constitutive mTOR activity negatively regulates glucose transporter trafficking. PMID- 18511521 TI - Glutamate availability is important in intramuscular amino acid metabolism and TCA cycle intermediates but does not affect peak oxidative metabolism. AB - Muscle glutamate is central to reactions producing 2-oxoglutarate, a tricarboxylic acid (TCA) cycle intermediate that essentially expands the TCA cycle intermediate pool during exercise. Paradoxically, muscle glutamate drops approximately 40-80% with the onset of exercise and 2-oxoglutarate declines in early exercise. To investigate the physiological relationship between glutamate, oxidative metabolism, and TCA cycle intermediates (i.e., fumarate, malate, 2 oxoglutarate), healthy subjects trained (T) the quadriceps of one thigh on the single-legged knee extensor ergometer (1 h/day at 70% maximum workload for 5 days/wk), while their contralateral quadriceps remained untrained (UT). After 5 wk of training, peak oxygen consumption (VO2peak) in the T thigh was greater than that in the UT thigh (P<0.05); VO2peak was not different between the T and UT thighs with glutamate infusion. Peak exercise under control conditions revealed a greater glutamate uptake in the T thigh compared with rest (7.3+/-3.7 vs. 1.0+/ 0.1 micromol.min(-1).kg wet wt(-1), P<0.05) without increase in TCA cycle intermediates. In the UT thigh, peak exercise (vs. rest) induced an increase in fumarate (0.33+/-0.07 vs. 0.02+/-0.01 mmol/kg dry wt (dw), P<0.05) and malate (2.2+/-0.4 vs. 0.5+/-0.03 mmol/kg dw, P<0.05) and a decrease in 2-oxoglutarate (12.2+/-1.6 vs. 32.4+/-6.8 micromol/kg dw, P<0.05). Overall, glutamate infusion increased arterial glutamate (P<0.05) and maintained this increase. Glutamate infusion coincided with elevated fumarate and malate (P<0.05) and decreased 2 oxoglutarate (P<0.05) at peak exercise relative to rest in the T thigh; there were no further changes in the UT thigh. Although glutamate may have a role in the expansion of the TCA cycle, glutamate and TCA cycle intermediates do not directly affect VO2peak in either trained or untrained muscle. PMID- 18511522 TI - Influence of priming exercise on pulmonary O2 uptake kinetics during transitions to high-intensity exercise from an elevated baseline. AB - It has been suggested that the slower O2 uptake (VO2) kinetics observed when exercise is initiated from an elevated baseline metabolic rate are linked to an impairment of muscle O2 delivery. We hypothesized that "priming" exercise would significantly reduce the phase II time constant (tau) during subsequent severe intensity cycle exercise initiated from an elevated baseline metabolic rate. Seven healthy men completed exercise transitions to 70% of the difference between gas exchange threshold (GET) and peak VO2 from a moderate-intensity baseline (90% GET) on three occasions in each of the "unprimed" and "primed" conditions. Pulmonary gas exchange, heart rate, and the electromyogram of m. vastus lateralis were measured during all tests. The phase II VO2 kinetics were slower when severe exercise was initiated from a baseline of moderate exercise compared with unloaded pedaling (mean+/-SD tau, 42+/-15 vs. 33+/-8 s; P<0.05), but were not accelerated by priming exercise (42+/-17 s; P>0.05). The amplitude of the VO2 slow component and the change in electromyogram from minutes 2 to 6 were both significantly reduced following priming exercise (VO2 slow component: from 0.47+/ 0.09 to 0.27+/-0.13 l/min; change in integrated electromyogram between 2 and 6 min: from 51+/-35 to 26+/-43% of baseline; P<0.05 for both comparisons). These results indicate that the slower phase II VO2 kinetics observed during transitions to severe exercise from an elevated baseline are not altered by priming exercise, but that the reduced VO2 slow component may be linked to changes in muscle fiber activation. PMID- 18511523 TI - Modulation of {beta}-adrenoceptor signaling in the hearts of 4-wk simulated weightlessness rats. AB - The modulation of beta-adrenoceptor signaling in the hearts of hindlimb unweighting (HU) simulated weightlessness rats has not been reported. In the present study, we adopted the rat tail suspension for 4 wk to simulate weightlessness; then the effects of simulated microgravity on beta-adrenoceptor signaling were studied. Mean arterial blood pressure (ABP), left ventricular pressure (LVP), systolic function (+dP/dtmax), and diastolic function (-dP/dtmax) were monitored in the course of the in vivo experiment. Single rat ventricular myocyte was obtained by the enzymatic dissociation method. Hemodynamics, myocyte contraction, and cAMP production in response to beta-adrenoceptor stimulation with isoproterenol or adenylyl cyclase stimulation with forskolin were measured, and Gs protein was also determined. Compared with the control group, no significant changes were found in heart weight, body weight and ABP, while LVP and +/-dP/dtmax were significantly reduced. The ABP decrease, LVP increase, and +/-dP/dtmax in response to isoproterenol administration were significantly attenuated in the HU group. The effects of isoproterenol on electrically induced single-cell contraction and cAMP production in myocytes of ventricles in the HU rats were significantly attenuated. The biologically active isoform, Gsalpha (45 kDa) in the heart, was unchanged. Both the increased electrically induced contraction and cAMP production in response to forskolin were also significantly attenuated in the simulated weightlessness rats. Above results indicated that impaired function of adenylyl cyclase causes beta-adrenoceptor desensitization, which may be partly responsible for the depression of cardiac function. PMID- 18511524 TI - Classifying individuals as physiological responders using hierarchical modeling. AB - We outline the use of hierarchical modeling for inference about the categorization of subjects into "responder" and "nonresponder" classes when the true status of the subject is latent (hidden). If uncertainty of classification is ignored during analysis, then statistical inference may be unreliable. An important advantage of hierarchical modeling is that it facilitates the correct modeling of the hidden variable in terms of predictor variables and hypothesized biological relationships. This allows researchers to formalize inference that can address questions about why some subjects respond and others do not. We illustrate our approach using a recent study of hepcidin excretion in female marathon runners (Roecker L, Meier-Buttermilch R, Brechte L, Nemeth E, Ganz T. Eur J Appl Physiol 95: 569-571, 2005). PMID- 18511525 TI - FKBP12 deficiency reduces strength deficits after eccentric contraction-induced muscle injury. AB - Strength deficits associated with eccentric contraction-induced muscle injury stem, in part, from excitation-contraction uncoupling. FKBP12 is a 12-kDa binding protein known to bind to the skeletal muscle sarcoplasmic reticulum Ca2+ release channel [ryanodine receptor (RyR1)] and plays an important role in excitation contraction coupling. To assess the effects of FKBP12 deficiency on muscle injury and recovery, we measured anterior crural muscle (tibialis anterior and extensor digitorum longus muscles) strength in skeletal muscle-specific FKBP12-deficient and wild-type (WT) mice before and after a single bout of 150 eccentric contractions, as well as before and after the performance of six injury bouts. Histological damage of the tibialis anterior muscle was assessed after injury. Body weight and peak isometric and eccentric torques were lower in FKBP12 deficient mice compared with WT mice. There were no differences between FKBP12 deficient and WT mice in preinjury peak isometric and eccentric torques when normalized to body weight, and no differences in the relative decreases in eccentric torque with a single or multiple injury bouts. After a single injury bout, FKBP12-deficient mice had less initial strength deficits and recovered faster (especially females) than WT mice, despite no differences in the degree of histological damage. After multiple injury bouts, FKBP12-deficient mice recovered muscle strength faster than WT mice and exhibited significantly less histological muscle damage than WT mice. In summary, FKBP12 deficiency results in less initial strength deficits and enhanced recovery from single (especially females) and repeated bouts of injury than WT mice. PMID- 18511526 TI - Habitual physical activity in daily life correlates positively with markers for mitochondrial capacity. AB - Physical exercise training is a powerful tool to maintain or improve mitochondrial density and function (mitochondrial capacity). This study aims to determine whether mitochondrial capacity is also associated with habitual physical activity in daily life (PADL). The capacity of classic markers for mitochondrial density, i.e., the capacity of citrate synthase (CS) and succinate dehydrogenase (SDH), as well the capacity of cytochrome c oxidase (COX) and beta hydroxyacyl-CoA dehydrogenase (HAD), was determined in homogenized muscle biopsy samples obtained from the vastus lateralis muscle of nonexercising healthy young (age 20+/-2 yr) subjects (31 women, 7 men). PADL was measured during two periods of 14 days using a triaxial accelerometer for movement registration. CS, SDH, and COX were positively associated with PADL [P<0.05, R=0.36, 95% confidence interval (CI): 1.3.10(-4) to 2.2.10(-3); P<0.05, R=0.39, 95% CI: 1.1.10(-5) to 9.9.10(-5); and P<0.05, R=0.33, 95% CI: 7.5.10(-6) to 3.6.10(-4), respectively], and HAD tended to correlate positively with PADL (P=0.06, R=0.31, 95% CI: -2.2.10(-5) to 1.1.10(-3)). The population was subsequently stratified based on the intensity of the activities performed. CS was only associated with PADL in subjects spending more time on high-intensity physical activity, whereas HAD was only associated with PADL in subjects spending less time on low intensity physical activity. We are the first to report that even within the range of normal daily life activities, mitochondrial capacity is positively associated with the level of habitual physical activity in daily life. Thus an active lifestyle may help to maintain or improve mitochondrial capacity. PMID- 18511527 TI - Effects of buspirone on posthypoxic ventilatory behavior in the C57BL/6J and A/J mouse strains. AB - Buspirone, a partial agonist of the serotonergic 5-HT1A receptor, improves breathing irregularities in humans with Rett syndrome or brain stem injury. The purpose of this study was to examine whether buspirone alters posthypoxic ventilatory behavior in C57BL/6J (B6) and A/J mouse strains. Measurements of ventilatory behavior were collected from unanesthetized adult male mice (n=6 for each strain) using the plethysmographic method. Mice were given intraperitoneal injections of vehicle or several doses of buspirone and exposed to 2 min of hypoxia (10% O2) followed by rapid reoxygenation (100% O2). Twenty minutes later, mice were tested for hypercapnic response (8% CO(2)-92% O2). On a separate day, mice were injected with the 5-HT1A receptor antagonist 4-iodo-N-{2-[4 (methoxyphenyl)-1-piperazinyl] ethyl}-N-2-pyridinylbenzamide (p-MPPI) before the injection of buspirone, and measurements were repeated. In separate studies, arterial blood-gas analysis was performed for each strain (n=12 in B6 and 10 in A/J) with buspirone or vehicle. In both strains, buspirone stimulated ventilation at rest. In the B6 mice, the hypoxic response was unchanged, but the response to hypercapnia was reduced with buspirone (5 mg/kg; P<0.05). With reoxygenation, vehicle-treated B6 exhibited periodic breathing and greater variation in ventilation compared with A/J (P<0.01). In B6 animals, >or=3 mg/kg of buspirone reduced variation and prevented the occurrence of posthypoxic periodic breathing. Both effects were reversed by p-MPPI. Treatment effect of buspirone was not explained by a difference in resting arterial blood gases. We conclude that buspirone improves posthypoxic ventilatory irregularities in the B6 mouse through its agonist effects on the 5-HT1A receptor. PMID- 18511528 TI - Influence of combined exercise training on indices of obesity, diabetes and cardiovascular risk in type 2 diabetes patients. AB - OBJECTIVE: To investigate the influence of combined exercise training on indices of obesity, diabetes and cardiovascular risk in type 2 diabetes patients. DESIGN: A double-blind randomized controlled trial with patients receiving either combination (COM), endurance (END) or no training (C). SETTING: Sint-Jozef hospital (Belgium), general practice (Holland). SUBJECTS: Forty-six type 2 diabetes patients (17 female, 29 male). INTERVENTIONS: COM versus END and C. Patients exercised for three months, three times a week for 1 hour. MAIN MEASURES: Six-minute walk test (6MW T), peak Vo(2), strength in upper and lower limbs, sit-to-stand, height, weight, body mass index, fat mass, glycosylated haemoglobin (HbA1c), glycaemia, triglycerides, high-density lipoprotein (HDL), total cholesterol and quality of life (General Health Survey Short Form (SF-36)). RESULTS: COM had significant better results on sit-to-stand (P<0.05), 6MW T (P<0.01), strength in upper (P<0.001) and lower limbs (P<0.001) compared with C. A different evolution among COM and C was found for HbA1c (P<0.05) and cholesterol (P<0.01), both decreased in COM and increased in C. HDL increased in COM and decreased in C (P<0.01). END had significant higher results on the 6MW T (P<0.01) compared with C. Compared with END, COM had significantly higher results on strength in upper (P<0.01) and lower limbs (P<0.01). The evolution of SF-36 items was not significantly different between the three groups. CONCLUSION: In diabetes type 2 patients, COM had significant better effects on indices of physical condition, diabetes and cardiovascular risk compared with C. Compared with END, COM gave a tendency towards better results, however more research with a larger number of participants is needed. PMID- 18511529 TI - Group exercise can improve participants' mobility in an outpatient rehabilitation setting: a randomized controlled trial. AB - OBJECTIVE: To establish the effects of group exercise on mobility and strength. DESIGN: Randomized controlled trial. SETTING: Two public hospital outpatient rehabilitation services. PARTICIPANTS: One hundred and seventy-three people (mean age 74.9 years, SD 10.8) with impaired mobility were randomized and 159 people (92%) completed the trial. INTERVENTIONS: Five-week, twice-weekly ;circuit-style' group exercise programme run by a physiotherapist (n = 85) and a no-intervention waiting list control group (n = 88). MAIN OUTCOME MEASURES: Three aspects of mobility: balance while standing and stepping (Step Test, semi-tandem and tandem stance times); sit-to-stand ability (rate and minimum height) and gait (6-metre and 6-minute walk tests). Lower limb muscle strength (knee flexion and extension). RESULTS: At retest, exercise participants had improved significantly more than their control counterparts on measures of balance while stepping, sit to stand and gait. Exercise participants averaged 1.6 more steps on the 15-second Step Test (95% confidence interval (CI) 0.5 to 2.8, P=0.005), walked an average of 0.12 m/s faster (95% CI 0.05 to 0.2, P=0.002) and took 2.5 fewer steps in 6 metres (95% CI -4.2 to -0.8, P=0.004). Exercise participants also averaged 0.04 more sit-to-stands/second, (95% CI 0.003 to 0.08, P=0.037) and walked an average of 30.9 metres further in 6 minutes (95% CI 9.4 to 52.4, P=0.005). There were no clinically important or statistically significant between-group differences at retest for the measures of strength (knee extension and flexion), balance while standing or minimal sit-to-stand height. CONCLUSION: This short-duration circuit class programme improved mobility, but not strength. PMID- 18511530 TI - Effectiveness of corticosteroid injection in adhesive capsulitis. AB - OBJECTIVE: To assess whether intraarticular corticosteroids improve the outcome of a comprehensive home exercise programme in patients with adhesive capsulitis. SETTING: The study was undertaken in the Physical Therapy and Rehabilitation Department of a Ministry of Health hospital in Turkey. SUBJECTS: Eighty patients with adhesive capsulitis were enrolled in the study. INTERVENTIONS: The patients were randomly assigned to two groups: Group 1 patients were given intraarticular corticosteroid (1 mL, 40 mg methylprednisolone acetate) followed by a 12-week comprehensive home exercise programme. Group 2 patients were given intraarticular serum physiologic (1 mL solution of 0.9% sodium chloride) followed by a 12-week comprehensive home exercise programme. MAIN MEASURES: The outcome parameters were Shoulder Pain and Disability Index and University of California-Los Angeles end result scores, night pain and shoulder passive range of motion. RESULTS: Mean actual changes in abduction range of motion, Shoulder Pain and Disability Index total score and Shoulder Pain and Disability Index-pain score were statistically different between the two groups at the second week, with the better scores determined in group 1. However, there were no significant differences between the groups at the 12th week. Medians of University of California-Los Angeles scores in the second week were significantly different between the two groups (P = 0.02), with better scores in group 1; however, the difference in 12th week scores was insignificant. CONCLUSIONS: Intraarticular corticosteroids have the additive effect of providing rapid pain relief, mainly in the first weeks of the exercise treatment period. In patients with adhesive capsulitis who have pain symptom predominantly, intraarticular corticosteroid therapy could be advised concomitantly with exercise. PMID- 18511531 TI - A prospective randomized controlled study of the role of botulinum toxin in whiplash-associated disorder. AB - OBJECTIVES: To investigate the effectiveness of botulinum toxin in preventing the development of chronic whiplash-associated disorder. DESIGN: Prospective, randomized, placebo-controlled double-blind study. SETTING: Regional Neurological Rehabilitation Centre with participants being at home. SUBJECTS: Thirty-seven patients with whiplash-associated disorder who remained symptomatic two months after injury. INTERVENTIONS: Patients were randomized to receive either 250 units botulinum toxin type A (Dysport) or placebo (normal saline). Four trigger points were injected with 0.625 mL of injectant. OUTCOME MEASURES: Tenderness to palpation scores, visual analogue pain scale, Vernon-Mior Neck Pain and Disability Index and cervical range of motion. Follow-up assessments were carried out at four weeks and three months after treatment. RESULTS: Twenty participants received botulinum toxin and 17 received placebo. Both groups showed a tendency towards improvement in pain scores, Vernon-Mior Index and range of motion at four weeks and three months, with the changes being more pronounced in the toxin group. The change in Vernon-Mior Index in the toxin group was both statistically and clinically significant (i.e. a change of score of > or = 5 from baseline to follow-up). Group comparisons did not meet statistical significance. CONCLUSION: The improvements in outcome measures suggest that botulinum toxin type A may have a role to play in the management of whiplash-associated disorder but larger studies are required to clarify the situation. PMID- 18511532 TI - Handle size as a task constraint in spoon-use movement in patients with Parkinson's disease. AB - OBJECTIVE: To examine the effect of spoon-handle size on kinematic performance in people with Parkinson's disease. DESIGN: A counterbalanced repeated-measures design. SETTING: A motor control laboratory in a university setting. SUBJECTS: Eighteen individuals with Parkinson's disease and 18 age-matched controls. EXPERIMENTAL CONDITIONS: Each participant was instructed to scoop water (simulated soup) using spoons with three different-sized handles. MAIN MEASURES: Kinematic variables (movement time, peak velocity and number of movement units) of arm movement, size of hand aperture and number of fingers to grasp the spoon. RESULTS: The movement of the participants with Parkinson's disease was faster (shorter movement time) and smoother (fewer movement units) when they used spoons with a small- or medium-sized handle than when using a spoon with a large-sized handle. In contrast, the healthy controls showed no significant differences in movement kinematics between handle sizes. Moreover, the participants with Parkinson's disease had a significantly smaller hand aperture and used more fingers to hold the spoons than the controls did. CONCLUSIONS: These results suggest that, for people with Parkinson's disease, a small-to-medium-sized handle is more suitable than a large-sized built-up handle. PMID- 18511533 TI - Sick leave reductions from a comprehensive manual therapy programme for low back pain: the Gotland Low Back Pain Study. AB - OBJECTIVE: To evaluate if a comprehensive manual therapy programme reduces sick leave due low back pain and facilitates return to work more than the conventional optimized activating care. DESIGN: A randomized controlled trial over a 10-week period with a two-year follow-up. SETTING: Primary health care and Visby Hospital, Municipality of Gotland, Sweden. SUBJECTS: One hundred and sixty patients (70 women, 90 men, ages 20-55 years) with acute or subacute low back pain with or without pain radiation into the legs. INTERVENTIONS: Standardized optimized activating care (n = 71) versus a comprehensive pragmatic manual therapy programme including specific corticosteroid injections (n = 89). MAIN MEASURES: Sick leave measured as net sick leave volume, point prevalence and return to work. RESULTS: After 10 weeks, significantly more manual therapy patients than reference patients had returned to work (hazards ratio 1.62, 95% confidence interval (CI) 1.006-2.60, P<0.05), and among those on sick leave at baseline, significantly fewer were still on sick leave (8/58 versus 13/40, ratio 0.35, 95% CI 0.13-0.97, P<0.05). For all other measures there were inconclusive differences in favour of the manual therapy group. No significant differences remained after two years. CONCLUSIONS: The manual therapy programme used in this study decreased sick leave and increased return to work more than the standardized optimized activating care only up to 10 weeks but not up to two years. PMID- 18511534 TI - Measures for rating social participation in people with aphasia: a systematic review. AB - BACKGROUND: Re-establishing participation in social life is an important aim of rehabilitation, but instruments to measure participation in people with aphasia are rare. AIMS: To identify and describe measures of social participation that may be specifically useful when measuring participation in people with aphasia. METHODS AND PROCEDURES: A systematic review of the literature concerning participation instruments was conducted. Then six speech and language therapists evaluated the suitability of selected participation measures for use in people with aphasia and a systematic literature review concerning the feasibility, internal consistency, validity, reliability and responsiveness of the measures selected by the therapists was carried out. RESULTS: In total 12 instruments measuring aspects of participation were found: seven measured actual performance and five measured actual performance combined with experienced problems. Two were considered unsuitable for people with aphasia, leaving 10. Six speech and language therapists working with people with aphasia scored the 10 selected instruments, and two instruments were judged as possibly suitable for use in people with aphasia: the Community Integration Questionnaire (CIQ) and the Nottingham Extended Activities of Daily Living (NEADL). However, the Community Integration Questionnaire is much closer to the concept of participation. The literature review concerning the psychometric properties of the Community Integration Questionnaire revealed that very little is known about the use of this instrument in people with aphasia. CONCLUSION: The Community Integration Questionnaire is possibly suitable for use in people with aphasia when measuring participation, but data on its psychometric properties in people with aphasia are absent. PMID- 18511535 TI - Mobility beyond the clinic: the effect of environment on gait and its measurement in community-ambulant stroke survivors. AB - OBJECTIVE: To explore the impact of a complex community environment on gait parameters (speed, step length and cadence) for community-dwelling participants with a previous stroke, and compare outcome measures commonly used in a clinical environment. DESIGN: Repeated measurement of participants in different environments. SETTING: One clinic and two community environments (suburban street and shopping mall). SUBJECTS: Thirty community-dwelling stroke participants with chronic stroke who were classified according to gait speed (20-50 m/min on 10 metre timed walk) as marginal community walkers. OUTCOME MEASURES: During a six minute walk test (6MWT) a step activity monitor (SAM) and odometer were used to calculate gait speed, step length and cadence. The 10-metre timed walk (10MTW) was measured in a clinic environment. ANALYSIS: A mixed linear model examined differences in gait measurements in the different environments. Bland-Altman analysis illustrated agreement between gait speed measures (6MWT and 10MTW). RESULTS: A statistically significant, but not a clinically significant difference in gait speed between some environments was found. Gait speed was slowest in the mall and fastest in the street with a difference of only 2.1 m/min between these environments (95% confidence interval (CI) -3.8 to -0.5, P<0.01). Comparison of clinic 10MTW and street 6MWT showed wide limits of agreement (-18.5 to 16.9 m/min) which improved for clinic 6MWT and street 6MWT comparisons (-5.7 to 8.9 m/min). CONCLUSION: Despite residual gait deficit, the gait parameters of these chronic stroke survivors did not deteriorate markedly under challenging conditions. The 6MWT is recommended as a clinical measure for community ambulation. PMID- 18511536 TI - The Pain Attitudes and Beliefs Scale for Physiotherapists: psychometric properties of the German version. AB - OBJECTIVES: To evaluate the psychometric properties of the German version of the Pain Attitudes and Beliefs Scale for Physiotherapists (PABS-PT-G). The original Dutch version is a 36-item self-report scale discriminating between a biomedical and a biopsychosocial orientation of therapists with regard to low back pain management. METHODS: The German version was generated by use of a forward backward translation procedure. In a cross-sectional study with a repeated measurement after five weeks, a total of 424 physiotherapists got a questionnaire package with the PABS-PT-G as well as the Tampa Scale of Kinesiophobia and patient vignettes. Item and factor analyses served to determine the psychometric properties of the scale. RESULTS: A total of 280 physiotherapists (response rate 79%) completed the questionnaires and a principal component analysis confirmed the two subscales of the original Dutch version. Internal consistency (Cronbach's alpha) of the items belonging to the biomedical factor amounted to 0.77 and to 0.58 for the items of the biopsychosocial factor respectively. Retest reliability showed a coefficient of 0.83 for the biomedical scale and of 0.70 for the biopsychosocial scale. Validity of both subscales of the PABS-PT-G was supported by statistically significant (P<0.01) and substantial correlations with the Tampa Scale of Kinesiophobia and with attitudes of the physiotherapists measured by patient vignettes (r between 0.37 and 0.72). CONCLUSION: The PABS-PT-G appears to be a reliable and valid instrument, which is equivalent to the original Dutch version. Further research is proposed to improve the reliability of the biopsychosocial subscale. PMID- 18511538 TI - Early events in protein folding: Is there something more than hydrophobic burst? AB - The presence of native contacts in the denatured state of many proteins suggests that elements of the biologically active structure of these molecules are formed during the initial stage of the folding process. The rapidity with which these events take place makes it difficult to study them in vitro, but, by the same token, suitable for studies in silico. With the help of all-atom, explicit solvent, molecular dynamics simulations we have followed in time, starting from elongated structureless conformations, the early events in the folding of src-SH3 domain and of proteins G, L, and CI2. It is observed that within the first 50 ns two important events take place, essentially independent of each other: hydrophobic collapse and formation of a few selected native contacts. The same contacts are also found in simulations carried out in the presence of guanidinium chloride in order to reproduce the conditions used to characterize experimentally the denatured state and testify to the fact that these contacts are to be considered a resilient characterizing property of the denaturated state. PMID- 18511537 TI - The structure of an archaeal homodimeric ligase which has RNA circularization activity. AB - The genome of Pyrococcus abyssi contains two open reading frames encoding proteins which had been previously predicted to be DNA ligases, Pab2002 and Pab1020. We show that while the former is indeed a DNA ligase, Pab1020 had no effect on the substrate deoxyoligo-ribonucleotides tested. Instead, Pab1020 catalyzes the nucleotidylation of oligo-ribonucleotides in an ATP-dependent reaction, suggesting that it is an RNA ligase. We have solved the structure of Pab1020 in complex with the ATP analog AMPPNP by single-wavelength anomalous dispersion (SAD), elucidating a structure with high structural similarity to the catalytic domains of two RNA ligases from the bacteriophage T4. Additional carboxy-terminal domains are also present, and one of these mediates contacts with a second protomer, which is related by noncrystallographic symmetry, generating a homodimeric structure. These C-terminal domains are terminated by short domain swaps which themselves end within 5 A of the active sites of the partner molecules. Additionally, we show that the protein is indeed capable of circularizing RNA molecules in an ATP-dependent reaction. These structural and biochemical results provide an insight into the potential physiological roles of Pab1020. PMID- 18511539 TI - Open clinical study of eye-drops containing tetrapeptides derived from substance P and insulin-like growth factor-1 for treatment of persistent corneal epithelial defects associated with neurotrophic keratopathy. AB - BACKGROUND/AIMS: Loss of corneal sensation results in the development of persistent corneal epithelial defects. The combination of a substance P-derived peptide (FGLM-amide) and an insulin-like growth factor-1 (IGF-1)-derived peptide (SSSR) stimulates rabbit corneal epithelial migration in vitro and rabbit corneal epithelial wound closure in vivo. The clinical efficacy of eye-drops containing FGLM-amide and SSSR for the treatment of persistent corneal epithelial defects in individuals with neurotrophic keratopathy was examined in a prospective open study. METHODS: Twenty-five consecutive patients (26 eyes) with persistent corneal epithelial defects associated with neurotrophic keratopathy were treated by administration of eye-drops containing FGLM-amide and SSSR. The course of epithelial healing was monitored by slit-lamp examination. RESULTS: Epithelial defects resurfaced completely in 19 of the 26 eyes (73%) within 4 weeks after treatment initiation. Complete resurfacing of epithelial defects was apparent in 18 of 22 (82%) or in one of four (25%) eyes without or with limbal stem cell deficiency, respectively. No adverse effects of treatment were observed in any subject. CONCLUSION: Eye-drops containing FGLM-amide and SSSR induced the rapid resurfacing of persistent epithelial defects in stem cell-positive individuals with neurotrophic keratopathy. PMID- 18511540 TI - The effect of travoprost on daytime intraocular pressure in normal tension glaucoma: a randomised controlled trial. AB - BACKGROUND/AIMS: To determine the medium-term effect of travoprost on the daytime intraocular pressure (IOP) of patients with normal tension glaucoma (NTG) METHODS: Newly diagnosed NTG patients underwent baseline, daytime, hourly IOP phasing. Patients were randomised to either treatment or no treatment (control). Treatment comprised once daily topical travoprost 0.004%. After 6 months, the participants underwent their second IOP phasing. RESULTS: Data from 88 participants were analysed-54 were randomised to treatment and 34 to the control group. The mean duration of treatment was 6 months. The average, maximum and minimum diurnal IOPs for treated patients were statistically significantly lower than for control patients at follow-up (p<0.001). When compared with baseline IOP, the travoprost treated group demonstrated a decrease of 16.1%, 13.5% and 16.7% in the average IOP, maximum IOP, and minimum IOP respectively. Of those treated, about one-third achieved a decrease in average IOP of at least 20%; only about one-tenth achieved a reduction of at least 30%. CONCLUSION: Travoprost monotherapy had a sustained hypotensive effect in NTG and achieved a reasonable or good response (>20% reduction in average IOP) in 32.9% of treated eyes. However, in the majority of eyes with NTG, travoprost monotherapy appeared unable to produce the desirable 30% reduction in average IOP. PMID- 18511541 TI - Visual rehabilitation in end-stage inflammatory ocular surface disease with the osteo-odonto-keratoprosthesis: results from the UK. AB - AIMS: To report the long-term results of osteo-odonto-keratoprosthesis (OOKP) surgery in the visual rehabilitation of patients with corneal blindness from end stage inflammatory ocular surface disease. METHODS: A non-comparative retrospective case series of 36 consecutive patients treated at the National OOKP referral centre in Brighton, UK, between November 1996 and March 2006. RESULTS: A total of 36 patients, with age ranging from 19 to 87 years (mean 51 (SD 19) years), were included in the analysis. The main preoperative diagnoses were Stevens-Johnson syndrome (n = 16, or 44%), severe thermal or chemical burns (n = 6, or 17%), and mucous membrane pemphigoid (n = 5, or 14%). The remainder of the cases comprised miscellaneous causes of dry eye (n = 9, or 25%), which included one each of graft versus host disease, ectodermal dysplasia, ionising radiation damage, cicatrising conjunctivitis from topical medication, trachoma, congenital trigeminal nerve hypoplasia, linear IgA disease, Sjogren syndrome and nutritional deficiency. Follow-up ranged from 6 months to 9 years (mean 3.9 (SD 2.5) years). Anatomical retention during the entirety of the follow-up period was seen in 72% of patients. The main factor resulting in anatomical failure was resorption of the OOKP lamina, which occurred in seven cases (or 19%). Predicted resorption in three cases resulted in successful planned exchange of the lamina, but two cases underwent emergency removal of the OOKP, and two cases developed endophthalmitis. Human leucocyte antigen-matched allografts suffered a higher rate of laminar resorption. Out of the entire cohort, 30 patients (or 83%) had some improvement in vision, 28 (or 78%) achieved vision of 6/60 or better, and 19 (or 53%) achieved 6/12 or better. The best-achieved vision was retained throughout the follow-up period in 61% of cases. Survival analysis suggested that the probability of retaining vision >6/60 5 years after surgery was 53 (10)%. Vision threatening complications occurred in nine cases (or 25%) and included endophthalmitis, retinal detachment and glaucoma. De novo glaucoma occurred in six patients (or 24%) but was seen overall in 17 patients (or 47%), 10 of whom required surgical treatment. CONCLUSION: OOKP surgery can restore useful and lasting vision in patients suffering from end-stage ocular surface disease, for whom conventional corneal surgery is not possible. The main problems seen in this study were laminar resorption, particularly in allografts, and glaucoma. PMID- 18511542 TI - Changes in meibomian fatty acids and clinical signs in patients with meibomian gland dysfunction after minocycline treatment. AB - AIMS: To assess the changes in ocular surface abnormalities and meibomian fatty acid composition in patients suffering from meibomian gland dysfunction (MGD) after treatment with oral minocycline associated with lid hygiene versus lid hygiene only. METHODS: We evaluated the break-up time, corneal staining and quality of meibomian excreta, and collected meibomian oil in 20 individuals suffering from MGD before and after 8 weeks of minocycline associated with lid hygiene (n = 10) or lid hygiene only (n = 10). Meibomian fatty acids were directly transmethylated and analysed by gas chromatography (GC) and GC mass spectrometry. RESULTS: The meibomian fatty acid composition was slightly modified after 8 weeks in both groups. The decrease in a branched-chain fatty acid (isoC20) was greater after minocycline treatment than after lid hygiene only ( 65% and -25%, respectively; p<0.05). Other fatty acids were unchanged. A significant improvement in the BUT was observed after minocycline treatment (p = 0.03). CONCLUSION: This study showed better tear film stability after minocycline treatment and a biological effect on meibomian fatty acid composition in MGD patients. Minocycline was more effective than lid hygiene alone. Both interventions partly corrected fatty acid composition abnormalities. Among the fatty acids, isoC20 could be a biological marker of MGD. PMID- 18511543 TI - VEGF-induced effects on proliferation, migration and tight junctions are restored by ranibizumab (Lucentis) in microvascular retinal endothelial cells. AB - BACKGROUND: Because vascular endothelial growth factor (VEGF) signalling is deregulated in diabetic retinopathy, the potential therapeutic effects of VEGF inhibitors such as the human VEGF-specific antibody ranibizumab are currently being tested. A study was undertaken to determine whether VEGF-stimulated processes in retinal endothelial cells are reversed by ranibizumab. METHODS: The influence of VEGF(121) and VEGF(165) on the proliferation and migration of immortalised bovine retinal endothelial cells (iBREC) was studied in the presence and absence of ranibizumab. In addition, the protein composition of tight junctions in the presence of VEGF and its inhibitor in iBREC was investigated. RESULTS: While both isoforms stimulated proliferation of iBREC, only VEGF(165) influenced cell migration. The addition of ranibizumab counteracted this stimulation without inhibition of the basal levels of migration and proliferation. Plasma membrane staining of the tight junction proteins occludin and claudin-1 disappeared in the presence of VEGF(165); there was no effect on claudin-5 and ZO-1 was only weakly affected. The addition of ranibizumab restored plasma membrane localisation of occludin and claudin-1. For claudin-1, the variation in total protein expression corresponded with the observed effects of VEGF(165) and ranibizumab. CONCLUSION: Ranibizumab reverses proliferation and cell migration stimulated by VEGF and delocalisation of tight junction proteins induced by VEGF(165) in iBREC. PMID- 18511544 TI - Work disability and health-related quality of life in males and females with psoriatic arthritis. AB - OBJECTIVES: To compare health status, demographic variables and work disability (WD) between males and females with psoriatic arthritis (PsA) in the 18-45 age group, and further to compare health status between those with and without WD for each gender and to identify variables associated with WD. METHODS: A cross sectional study was carried out of patients with PsA with peripheral arthritis at the time at which they started disease-modifying antirheumatic drug therapy (DMARD) and/or biological treatment. Patients receiving a permanent national WD pension corresponding to >or=50% were defined as work disabled. Gender differences were examined with regard to health status, demographic variables and WD. Mann-Whitney U test and Pearson chi(2) were applied for group comparisons between males and females and work disabled versus not work disabled for each gender. Multiple logistic regression analyses with adjustments for duration of education, disease duration, age, erosive disease, disability score (Modified Health Assessment Questionnaire; MHAQ), the short form-36 (SF-36) mental health score, and gender were used to identify variables associated with WD. RESULTS: Out of 271 (102 females) patients, the number (%) of work-disabled females/males was 33 (32.7%)/29 (17.4%) (p = 0.004). Work-disabled patients had generally worse health status than non-work-disabled patients, and these differences were generally more pronounced in males than in females. In the multiple logistic regression model, low educational level, increasing disability score (MHAQ), presence of erosive disease, female gender and disease duration were independently associated with WD. CONCLUSIONS: WD in patients with PsA below 45 years of age was independently associated with educational level, disability score, erosive disease, female gender and disease duration. PMID- 18511545 TI - Over-representation of construction-related occupations in male patients with systemic sclerosis. AB - AIMS: Based on preliminary observations, we tested the hypothesis that construction-related occupations are associated with systemic sclerosis (SSc). METHODS: The professional occupation of 91 patients with SSc (71 females and 20 males) was recorded. Categorisation into construction-related and other professions was performed. A double definition was used for construction-related occupations. The first (limited) definition was based upon categories of the Belgian National Institute of Statistics (NIS) occupational list. The following occupations were considered construction-related: electricians, joiners, masons and tilers, plumbers and pipefitters. The use of this list also allows us to compare the distribution of professions in these patients with that in the general population. As the NIS occupational list is limitative and leaves out some "real-life" construction-related occupations, a second and broader interpretation was given to the concept of construction-related occupations. RESULTS: The prevalence of construction-related professions in males with SSc, according to the limited definition, was 10-fold higher than in the general working population (50% vs 5%; p<0.001). Interestingly, most of the patients with construction-related occupations were electricians. In the broader interpretation, 75% of the men with SSc fell into the category of construction related occupations. CONCLUSIONS: The data show an association between SSc and professional occupation. PMID- 18511546 TI - Systemic sclerosis and its pulmonary complications in The Netherlands: an epidemiological study. AB - The prevalence and incidence of systemic sclerosis (SSc) in The Netherlands is unknown. The same holds true for its leading causes of death: pulmonary fibrosis and pulmonary arterial hypertension (PAH), for which effective treatment options have recently become available. OBJECTIVE: To establish the prevalence and incidence of SSc and its pulmonary complications. METHODS: Detailed information on patients in the POEMAS registry, "Pulmonary Hypertension Screening, a Multidisciplinary Approach in Scleroderma", consisting of 819 patients, was combined with a nationwide questionnaire. RESULTS: By combining the two sources the prevalence of SSc was found to be 8.9 per 100 000 adults. The incidence was 0.77 patients per 100 000 per year. PAH was diagnosed in 9.9% of SSc patients. The prevalence of interstitial lung disease in SSc varied from 19% to 47% depending on the definition used. CONCLUSION: This study clarifies the epidemiology of SSc in The Netherlands and confirms the frequent occurrence of pulmonary complications, based on 654 cases. This can and will be studied further in the ongoing POEMAS study. PMID- 18511547 TI - Measuring quality of care for rheumatic diseases using an electronic medical record. AB - OBJECTIVES: The objective of this study was twofold: (1) to determine how best to measure adherence with time-dependent quality indicators (QIs) related to laboratory monitoring, and (2) to assess the accuracy and efficiency of gathering QI adherence information from an electronic medical record (EMR). METHODS: A random sample of 100 patients were selected who had at least three visits with the diagnosis of rheumatoid arthritis (RA) at Brigham and Women's Hospital Arthritis Center in 2005. Using the EMR, it was determined whether patients had been prescribed a disease-modifying antirheumatic drug (DMARD) (QI #1) and if patients starting therapy received appropriate baseline laboratory testing (QI #2). For patients consistently prescribed a DMARD, adherence with follow-up testing (QI #3) was calculated using three different methods, the Calendar, Interval and Rolling Interval METHOD: . RESULTS: It was found that 97% of patients were prescribed a DMARD (QI #1) and baseline tests were completed in 50% of patients (QI #2). For follow-up testing (QI #3), mean adherence was 60% for the Calendar Method, 35% for the Interval Method, and 48% for the Rolling Interval Method. Using the Rolling Interval Method, adherence rates were similar across drug and laboratory testing type. CONCLUSIONS: Results for adherence with laboratory testing QIs for DMARD use differed depending on how the QIs were measured, suggesting that care must be taken in clearly defining methods. While EMRs will provide important opportunities for measuring adherence with QIs, they also present challenges that must be examined before widespread adoption of these data collection methods. PMID- 18511548 TI - Allelic variants of streptokinase from Streptococcus pyogenes display functional differences in plasminogen activation. AB - A common mammalian defense mechanism employed to prevent systemic dissemination of invasive bacteria involves occlusion of local microvasculature and encapsulation of bacteria within fibrin networks. Acquisition of plasmin activity at the bacterial cell surface circumvents this defense mechanism, allowing invasive disease initiation. To facilitate this process, S. pyogenes secretes streptokinase, a plasminogen-activating protein. Streptokinase polymorphism exhibited by S. pyogenes isolates is well characterized. However, the functional differences displayed by these variants and the biological significance of this variation has not been elucidated. Phylogenetic analysis of ska sequences from 28 S. pyogenes isolates revealed 2 main sequence clusters (clusters 1 and 2). All strains secreted streptokinase, as determined by Western blotting, and were capable of acquiring cell surface plasmin activity after incubation in human plasma. Whereas culture supernatants from strains containing cluster 1 ska alleles also displayed soluble plasminogen activation activity, supernatants from strains containing cluster 2 ska alleles did not. Furthermore, plasminogen activation activity in culture supernatants from strains containing cluster 2 ska alleles could only be detected when plasminogen was prebound with fibrinogen. This study indicates that variant streptokinase proteins secreted by S. pyogenes isolates display differing plasminogen activation characteristics and may therefore play distinct roles in disease pathogenesis. PMID- 18511549 TI - Phosphorylation of tau regulates its axonal transport by controlling its binding to kinesin. AB - Defective axonal transport has been proposed as an underlying mechanism that may give rise to neurodegeneration. We investigated the effect of phosphorylation on the axonal transport of tau, a neuronal protein that stabilizes microtubules and is hyperphosphorylated and mislocalized in Alzheimer's disease. We report here that specific inhibition of glycogen synthase kinase-3 (GSK-3) reduces tau phosphorylation and significantly decreases the overall rate of axonal transport of tau in rat cortical neurons. Tau mutants, with serine/threonine targets of GSK 3 mutated to glutamate to mimic a permanent state of phosphorylation, were transported at a significantly increased rate compared to wild-type tau. Conversely, tau mutants, in which alanine replaced serine/threonine to mimic permanent dephosphorylation, were transported at a decreased rate compared to wild-type tau. We also found that tau interacts with the light chain of kinesin-1 and that this is dependent on the phosphorylation state of tau. Tau phosphorylation by GSK-3 increased binding, and dephosphorylated tau exhibited a reduced association with kinesin-1. We conclude that GSK-3 phosphorylation of tau modulates its axonal transport by regulating binding to kinesin-1. Hyperphosphorylated tau in Alzheimer's disease appearing first in distal portions of axons may result from aberrant axonal transport of phosphorylated tau reported here. PMID- 18511550 TI - Dual role of the arginine methyltransferase CARM1 in the regulation of c-Fos target genes. AB - Fos proteins, the prototypic members of basic region-leucine zipper (bZIP) transcription factors, bind to other bZIP proteins to form the activator protein 1 (AP-1) complex, which regulates the expression of a plethora of target genes. Notably, c-Fos target genes include members of the matrix metalloproteinase (MMP) gene family and c-fos is overexpressed in a number of metastatic cancers, suggesting its direct involvement in this process. Here, we reveal that c-Fos mediated transcriptional activation is regulated by the protein arginine methyltransferase CARM1 and by all three members of the p160 protein family of coactivators. Carm1-deficient cells showed a dramatic reduction in the expression level of c-Fos target genes MMP-1b, -3, and -13, indicating a major role for CARM1 in regulating the expression of these genes. RNA interference combined with quantitative polymerase chain reaction demonstrated that CARM1 and p160 proteins synergize to activate expression of MMP-1b, -3, and -13 in vivo. Furthermore, we show that CARM1 also regulates MMP expression at the post-transcriptional level, either positively or negatively. Our data indicate that CARM1 can play a dual role in the expression of AP-1 target genes involved in cancer or other diseases by acting at the transcriptional as well as at the post-transcriptional levels. PMID- 18511551 TI - Human tumor nanoparticles induce apoptosis of pancreatic cancer cells. AB - Exosomes are vesicles secreted by most hematopoietic cells on fusion of multivesicular endosomes with the plasma membrane. Many studies have reported that exosomes may also be released by tumor cells. Exosomes are believed to play an antitumor role through immune cells. We asked whether tumor exosomes have biological activities on tumor cells. We report that human pancreatic tumor nanoparticles, exosome-like as characterized by proteomic analyses and rich in lipid rafts, decreased tumor cell proliferation. Nanoparticles increased Bax and decreased Bcl-2 expressions. Caspase-3 and -9 but not caspase-8 inhibitors impaired apoptosis, which implicates the mitochondria apoptotic pathway. The ceramide-sphingomyelin apoptotic pathway was inoperative. Moreover, nanoparticles induced phosphatase and tensin homolog (PTEN) and glycogen synthase kinase (GSK) 3beta activation and decreased pyruvate dehydrogenase activity. In nanoparticle treated cells, PTEN formed complexes with actin, beta-catenin, and GSK-3beta. Thus, beta-catenin may no longer be available to activate the survival pathway. Nanoparticles triggered the down-regulation of cyclin D1 and poly(ADP-ribose) polymerase. Hence, nanoparticles counteracted the constitutively activated phosphatidylinositol 3-kinase/Akt survival pathway to drive tumor cells toward apoptosis. Our study provides the first evidence of an apoptotic function of tumor-derived nanoparticles on tumor cells. We propose a new role for nanoparticles, i.e., as signal carriers for interaction between cells, which may have implications in physiopathological situations. PMID- 18511553 TI - Natural selection does not explain cultural rates of change. PMID- 18511552 TI - Impaired olfaction in mice lacking aquaporin-4 water channels. AB - Aquaporin-4 (AQP4) is a water-selective transport protein expressed in glial cells throughout the central nervous system. AQP4 deletion in mice produces alterations in several neuroexcitation phenomena, including hearing, vision, epilepsy, and cortical spreading depression. Here, we report defective olfaction and electroolfactogram responses in AQP4-null mice. Immunofluorescence indicated strong AQP4 expression in supportive cells of the nasal olfactory epithelium. The olfactory epithelium in AQP4-null mice had identical appearance, but did not express AQP4, and had approximately 12-fold reduced osmotic water permeability. Behavioral analysis showed greatly impaired olfaction in AQP4-null mice, with latency times of 17 +/- 0.7 vs. 55 +/- 5 s in wild-type vs. AQP4-null mice in a buried food pellet test, which was confirmed using an olfactory maze test. Electroolfactogram voltage responses to multiple odorants were reduced in AQP4 null mice, with maximal responses to triethylamine of 0.80 +/- 0.07 vs. 0.28 +/- 0.03 mV. Similar olfaction and electroolfactogram defects were found in outbred (CD1) and inbred (C57/bl6) mouse genetic backgrounds. Our results establish AQP4 as a novel determinant of olfaction, the deficiency of which probably impairs extracellular space K(+) buffering in the olfactory epithelium. PMID- 18511554 TI - Reply to Bagni: On BC1 RNA and the fragile X mental retardation protein. PMID- 18511556 TI - A drug-controllable tag for visualizing newly synthesized proteins in cells and whole animals. AB - Research on basic cellular processes involving local production or delivery of proteins, such as activity-dependent synaptic modification in neurons, would benefit greatly from a robust, nontoxic method to visualize selectively newly synthesized copies of proteins of interest within cells, tissues, or animals. We report a technique for covalent labeling of newly synthesized proteins of interest based on drug-dependent preservation of epitope tags. Epitope tags are removed from proteins of interest immediately after translation by the activity of a sequence-specific protease until the time a protease inhibitor is added, after which newly synthesized protein copies retain their tags. This method, which we call TimeSTAMP for time-specific tagging for the age measurement of proteins, allows sensitive and nonperturbative visualization and quantification of newly synthesized proteins of interest with exceptionally tight temporal control. We demonstrate applications of TimeSTAMP in retrospectively identifying growing synapses in cultured neurons and in visualizing the distribution of recently synthesized proteins in intact fly brains. PMID- 18511557 TI - Asymmetric mitosis: Unequal segregation of proteins destined for degradation. AB - Mitotic cell division ensures that two daughter somatic cells inherit identical genetic material. Previous work has shown that signaling by the Smad1 transcription factor is terminated by polyubiquitinylation and proteasomal degradation after essential phosphorylations by MAPK and glycogen synthase kinase 3 (GSK3). Here, we show that, unexpectedly, proteins specifically targeted for proteasomal degradation are inherited preferentially by one mitotic daughter during somatic cell division. Experiments with dividing human embryonic stem cells and other mammalian cultured cell lines demonstrated that in many supposedly equal mitoses the segregation of proteins destined for degradation (Smad1 phosphorylated by MAPK and GSK3, phospho-beta-catenin, and total polyubiquitinylated proteins) was asymmetric. Transport of pSmad1 targeted for degradation to the centrosome required functional microtubules. In vivo, an antibody specific for Mad phosphorylated by MAPK showed that this antigen was associated preferentially with one of the two centrosomes in Drosophila embryos at cellular blastoderm stage. We propose that this remarkable cellular property may be explained by the asymmetric inheritance of peripheral centrosomal proteins when centrioles separate and migrate to opposite poles of the cell, so that one mitotic daughter remains pristine. We conclude that many mitotic divisions are unequal, unlike what was previously thought. PMID- 18511558 TI - Understanding ligand-based modulation of the Hsp90 molecular chaperone dynamics at atomic resolution. AB - Molecular switching and ligand-based modulation of the 90-kDa heat-shock protein (Hsp90) chaperone activity may ultimately facilitate conformational coupling to the ATPase cycle along with activation and recruitment of the broad range of client proteins. We present an atomic resolution analysis of the Hsp90 N-terminal domain (NTD) binding energy landscape by simulating protein dynamics with a range of binding partners. We show that the activity of the molecular chaperone may be linked to (i) local folding-unfolding transitions and conformational switching of the "active site lid" upon binding and (ii) differences in the underlying protein dynamics as a function of the binding partner. This study suggests that structural plasticity of the Hsp90 NTD can be exploited by the molecular chaperone machinery to modulate enhanced structural rigidity during ATP binding and increased protein flexibility as a consequence of the inhibitor binding. The present study agrees with the experimental structural data and provides a plausible molecular model for understanding mechanisms of modulation of molecular chaperone activities by binding partners. PMID- 18511559 TI - Next generation of adeno-associated virus 2 vectors: point mutations in tyrosines lead to high-efficiency transduction at lower doses. AB - Recombinant adeno-associated virus 2 (AAV2) vectors are in use in several Phase I/II clinical trials, but relatively large vector doses are needed to achieve therapeutic benefits. Large vector doses also trigger an immune response as a significant fraction of the vectors fails to traffic efficiently to the nucleus and is targeted for degradation by the host cell proteasome machinery. We have reported that epidermal growth factor receptor protein tyrosine kinase (EGFR-PTK) signaling negatively affects transduction by AAV2 vectors by impairing nuclear transport of the vectors. We have also observed that EGFR-PTK can phosphorylate AAV2 capsids at tyrosine residues. Tyrosine-phosphorylated AAV2 vectors enter cells efficiently but fail to transduce effectively, in part because of ubiquitination of AAV capsids followed by proteasome-mediated degradation. We reasoned that mutations of the surface-exposed tyrosine residues might allow the vectors to evade phosphorylation and subsequent ubiquitination and, thus, prevent proteasome-mediated degradation. Here, we document that site-directed mutagenesis of surface-exposed tyrosine residues leads to production of vectors that transduce HeLa cells approximately 10-fold more efficiently in vitro and murine hepatocytes nearly 30-fold more efficiently in vivo at a log lower vector dose. Therapeutic levels of human Factor IX (F.IX) are also produced at an approximately 10-fold reduced vector dose. The increased transduction efficiency of tyrosine-mutant vectors is due to lack of capsid ubiquitination and improved intracellular trafficking to the nucleus. These studies have led to the development of AAV vectors that are capable of high-efficiency transduction at lower doses, which has important implications in their use in human gene therapy. PMID- 18511560 TI - An original adaptation of photosynthesis in the marine green alga Ostreococcus. AB - Adaptation of photosynthesis in marine environment has been examined in two strains of the green, picoeukaryote Ostreococcus: OTH95, a surface/high-light strain, and RCC809, a deep-sea/low-light strain. Differences between the two strains include changes in the light-harvesting capacity, which is lower in OTH95, and in the photoprotection capacity, which is enhanced in OTH95. Furthermore, RCC809 has a reduced maximum rate of O(2) evolution, which is limited by its decreased photosystem I (PSI) level, a possible adaptation to Fe limitation in the open oceans. This decrease is, however, accompanied by a substantial rerouting of the electron flow to establish an H(2)O-to-H(2)O cycle, involving PSII and a potential plastid plastoquinol terminal oxidase. This pathway bypasses electron transfer through the cytochrome b(6)f complex and allows the pumping of "extra" protons into the thylakoid lumen. By promoting the generation of a large DeltapH, it facilitates ATP synthesis and nonphotochemical quenching when RCC809 cells are exposed to excess excitation energy. We propose that the diversion of electrons to oxygen downstream of PSII, but before PSI, reflects a common and compulsory strategy in marine phytoplankton to bypass the constraints imposed by light and/or nutrient limitation and allow successful colonization of the open-ocean marine environment. PMID- 18511561 TI - A NOD2-NALP1 complex mediates caspase-1-dependent IL-1beta secretion in response to Bacillus anthracis infection and muramyl dipeptide. AB - NOD2, a NOD-like receptor (NLR), is an intracellular sensor of bacterial muramyl dipeptide (MDP) that was suggested to promote secretion of the proinflammatory cytokine IL-1beta. Yet, the molecular mechanism by which NOD2 can stimulate IL 1beta secretion, and its biological significance were heretofore unknown. We found that NOD2 through its N-terminal caspase recruitment domain directly binds and activates caspase-1 to trigger IL-1beta processing and secretion in MDP stimulated macrophages, whereas the C-terminal leucine-rich repeats of NOD2 prevent caspase-1 activation in nonstimulated cells. MDP challenge induces the association of NOD2 with another NLR protein, NALP1, and gel filtration analysis revealed the formation of a complex consisting of NOD2, NALP1, and caspase-1. Importantly, Bacillus anthracis infection induces IL-1beta secretion in a manner that depended on caspase-1 and NOD2. In vitro, Anthrax lethal toxin strongly potentiated IL-1beta secretion, and that response was NOD2 and caspase-1 dependent. Thus, NOD2 plays a key role in the B. anthracis-induced inflammatory response by being a critical mediator of IL-1beta secretion. PMID- 18511562 TI - ALIX-CHMP4 interactions in the human ESCRT pathway. AB - The ESCRT pathway facilitates membrane fission events during enveloped virus budding, multivesicular body formation, and cytokinesis. To promote HIV budding and cytokinesis, the ALIX protein must bind and recruit CHMP4 subunits of the ESCRT-III complex, which in turn participate in essential membrane remodeling functions. Here, we report that the Bro1 domain of ALIX binds specifically to C terminal residues of the human CHMP4 proteins (CHMP4A-C). Crystal structures of the complexes reveal that the CHMP4 C-terminal peptides form amphipathic helices that bind across the conserved concave surface of ALIX(Bro1). ALIX-dependent HIV 1 budding is blocked by mutations in exposed ALIX(Bro1) residues that help contribute to the binding sites for three essential hydrophobic residues that are displayed on one side of the CHMP4 recognition helix (M/L/IxxLxxW). The homologous CHMP1-3 classes of ESCRT-III proteins also have C-terminal amphipathic helices, but, in those cases, the three hydrophobic residues are arrayed with L/I/MxxxLxxL spacing. Thus, the distinct patterns of hydrophobic residues provide a "code" that allows the different ESCRT-III subunits to bind different ESCRT pathway partners, with CHMP1-3 proteins binding MIT domain-containing proteins, such as VPS4 and Vta1/LIP5, and CHMP4 proteins binding Bro1 domain-containing proteins, such as ALIX. PMID- 18511563 TI - Lithium monoxide anion: a ground-state triplet with the strongest base to date. AB - Lithium monoxide anion (LiO(-)) has been generated in the gas phase and is found to be a stronger base than methyl anion (CH(3)(-)). This makes LiO(-) the strongest base currently known, and it will be a challenge to produce a singly charged or multiply charged anion that is more basic. The experimental acidity of lithium hydroxide is DeltaH degrees (acid) = 425.7 +/- 6.1 kcal.mol(-1) (1 kcal = 4.184 kJ) and, when combined with results of high-level computations, leads to our best estimate for the acidity of 426 +/- 2 kcal.mol(-1). PMID- 18511564 TI - Inflationary dynamics for matrix eigenvalue problems. AB - Many fields of science and engineering require finding eigenvalues and eigenvectors of large matrices. The solutions can represent oscillatory modes of a bridge, a violin, the disposition of electrons around an atom or molecule, the acoustic modes of a concert hall, or hundreds of other physical quantities. Often only the few eigenpairs with the lowest or highest frequency (extremal solutions) are needed. Methods that have been developed over the past 60 years to solve such problems include the Lanczos algorithm, Jacobi-Davidson techniques, and the conjugate gradient method. Here, we present a way to solve the extremal eigenvalue/eigenvector problem, turning it into a nonlinear classical mechanical system with a modified Lagrangian constraint. The constraint induces exponential inflationary growth of the desired extremal solutions. PMID- 18511565 TI - A dicarboxylate/4-hydroxybutyrate autotrophic carbon assimilation cycle in the hyperthermophilic Archaeum Ignicoccus hospitalis. AB - Ignicoccus hospitalis is an anaerobic, autotrophic, hyperthermophilic Archaeum that serves as a host for the symbiotic/parasitic Archaeum Nanoarchaeum equitans. It uses a yet unsolved autotrophic CO(2) fixation pathway that starts from acetyl CoA (CoA), which is reductively carboxylated to pyruvate. Pyruvate is converted to phosphoenol-pyruvate (PEP), from which glucogenesis as well as oxaloacetate formation branch off. Here, we present the complete metabolic cycle by which the primary CO(2) acceptor molecule acetyl-CoA is regenerated. Oxaloacetate is reduced to succinyl-CoA by an incomplete reductive citric acid cycle lacking 2 oxoglutarate dehydrogenase or synthase. Succinyl-CoA is reduced to 4 hydroxybutyrate, which is then activated to the CoA thioester. By using the radical enzyme 4-hydroxybutyryl-CoA dehydratase, 4-hydroxybutyryl-CoA is dehydrated to crotonyl-CoA. Finally, beta-oxidation of crotonyl-CoA leads to two molecules of acetyl-CoA. Thus, the cyclic pathway forms an extra molecule of acetyl-CoA, with pyruvate synthase and PEP carboxylase as the carboxylating enzymes. The proposal is based on in vitro transformation of 4-hydroxybutyrate, detection of all enzyme activities, and in vivo-labeling experiments using [1 (14)C]4-hydroxybutyrate, [1,4-(13)C(2)], [U-(13)C(4)]succinate, or [1 (13)C]pyruvate as tracers. The pathway is termed the dicarboxylate/4 hydroxybutyrate cycle. It combines anaerobic metabolic modules to a straightforward and efficient CO(2) fixation mechanism. PMID- 18511566 TI - Thiolate-bridged dinuclear iron(tris-carbonyl)-nickel complexes relevant to the active site of [NiFe] hydrogenase. AB - The reaction of NiBr(2)(EtOH)(4) with a 1:2-3 mixture of FeBr(2)(CO)(4) and Na(SPh) generated a linear trinuclear Fe-Ni-Fe cluster (CO)(3)Fe(mu-SPh)(3)Ni(mu SPh)(3)Fe(CO)(3), 1, whereas the analogous reaction system FeBr(2)(CO)(4)/Na(S(t)Bu)/NiBr(2)(EtOH)(4) (1:2-3:1) gave rise to a linear tetranuclear Fe-Ni-Ni-Fe cluster [(CO)(3)Fe(mu-S(t)Bu)(3)Ni(mu-Br)](2), 2. By using this tetranuclear cluster 2 as the precursor, we have developed a new synthetic route to a series of thiolate-bridged dinuclear Fe(CO)(3)-Ni complexes, the structures of which mimic [NiFe] hydrogenase active sites. The reactions of 2 with SC(NMe(2))(2) (tmtu), Na{S(CH(2))(2)SMe} and ortho-NaS(C(6)H(4))SR (R = Me, (t)Bu) led to isolation of (CO)(3)Fe(mu-S(t)Bu)(3)NiBr(tmtu), 3, (CO)(3)Fe(S(t)Bu)(mu-S(t)Bu)(2)Ni{S(CH(2))(2)SMe}, 4, and (CO)(3)Fe(S(t)Bu)(mu S(t)Bu)(2)Ni{S(C(6)H(4))SR}, 5a (R = Me) and 5b (R = (t)Bu), respectively. On the other hand, treatment of 2 with 2-methylthio-phenolate (ortho-O(C(6)H(4))SMe) in methanol resulted in (CO)(3)Fe(mu-S(t)Bu)(3)Ni(MeOH){O(C(6)H(4))SMe}, 6a. The methanol molecule bound to Ni is labile and is readily released under reduced pressure to afford (CO)(3)Fe(S(t)Bu)(mu-S(t)Bu)(2)Ni{O(C(6)H(4))SMe}, 6b, and the coordination geometry of nickel changes from octahedral to square planar. Likewise, the reaction of 2 with NaOAc in methanol followed by crystallization from THF gave (CO)(3)Fe(mu-S(t)Bu)(3)Ni(THF)(OAc), 7. The dinuclear complexes, 3 7, are thermally unstable, and a key to their successful isolation is to carry out the reactions and manipulations at -40 degrees C. PMID- 18511567 TI - Cdx gene deficiency compromises embryonic hematopoiesis in the mouse. AB - Cdx genes (Cdx1, Cdx2, and Cdx4) encode a family of caudal-related transcription factors that mediate anterior-posterior patterning during embryogenesis through Hox gene regulation. Homologues in the zebrafish have been shown to play key roles in blood formation. To define the role of Cdx genes during embryonic hematopoiesis in a mammalian system, we examined the hematopoietic potential of Cdx-deficient mouse embryonic stem cells (ESCs) in vitro and in vivo. Individual Cdx-deficient ESCs exhibited impaired embryonic hematopoietic progenitor formation and altered Hox gene expression, most notably for Cdx2 deficiency. A more severe hematopoietic defect was observed with compound Cdx deficiency than loss of function of any single Cdx gene. Reduced hematopoietic progenitor formation of ESCs deficient in multiple Cdx genes could be rescued by ectopic expression of Cdx4, concomitant with partially restored Hox gene expression. These results reveal an essential and partially redundant role for multiple Cdx genes during embryonic hematopoiesis in the mouse. PMID- 18511568 TI - Coevolution at protein complex interfaces can be detected by the complementarity trace with important impact for predictive docking. AB - Protein surfaces are under significant selection pressure to maintain interactions with their partners throughout evolution. Capturing how selection pressure acts at the interfaces of protein-protein complexes is a fundamental issue with high interest for the structural prediction of macromolecular assemblies. We tackled this issue under the assumption that, throughout evolution, mutations should minimally disrupt the physicochemical compatibility between specific clusters of interacting residues. This constraint drove the development of the so-called Surface COmplementarity Trace in Complex History score (SCOTCH), which was found to discriminate with high efficiency the structure of biological complexes. SCOTCH performances were assessed not only with respect to other evolution-based approaches, such as conservation and coevolution analyses, but also with respect to statistically based scoring methods. Validated on a set of 129 complexes of known structure exhibiting both permanent and transient intermolecular interactions, SCOTCH appears as a robust strategy to guide the prediction of protein-protein complex structures. Of particular interest, it also provides a basic framework to efficiently track how protein surfaces could evolve while keeping their partners in contact. PMID- 18511569 TI - Mosaicism in sporadic neurofibromatosis type 1: variations on a theme common to other hereditary cancer syndromes? AB - Mosaicism constitutes a frequent complication of the genotype-phenotype relationship in genetic disease and is an important consideration for the estimation of transmission risk. Mosaicism has been identified in several hereditary cancer syndromes including retinoblastoma, familial adenomatous polyposis coli, von Hippel-Lindau disease and neurofibromatosis type 2. Recent data support the postulate that the frequency of mosaicism is increased in cancer predisposition syndromes characterised by high new mutation rates. Since the new mutation rate is very high in neurofibromatosis type 1 (NF1), mosaicism might reasonably be expected to be frequent among sporadic cases but this remains to be formally demonstrated. Here we summarise current knowledge of mosaicism in NF1, focusing on the types of mutations identified as well as their inferred developmental timing and representation in different cell types, and assess the potential impact of high frequency mosaicism on mutation screening in patients with apparent de novo NF1. PMID- 18511570 TI - Molecular basis of the Li-Fraumeni syndrome: an update from the French LFS families. AB - We have performed an extensive analysis of TP53 in 474 French families suggestive of Li-Fraumeni syndrome (LFS), including 232 families fulfilling the Chompret criteria. We identified a germline alteration of TP53 in 82 families (17%), in 67/232 of the families fulfilling the Chompret criteria (29%) and in 15/242 which did not fulfil these criteria (6%). Most of the alterations corresponded to missense mutations (67%), and we identified in four families genomic deletions removing the entire TP53 locus, the promoter and the non-coding exon 1, or exons 2-10. These results represent a definitive argument demonstrating that LFS results from TP53 haplodeficiency. The mean ages of tumour onset were significantly different between patients harbouring TP53 missense mutations and other types of alterations, missense mutations being associated with a 9 year earlier tumour onset. These results confirm that missense mutations not only inactivate p53 but also have an additional oncogenic effect. Germline alterations of TP53 that lead exclusively to loss of function are therefore associated with a later age of tumour onset and the presence of such mutations should be considered in atypical LFS families with tumours diagnosed after 40 years. PMID- 18511571 TI - A novel A121T mutation in human cationic trypsinogen associated with hereditary pancreatitis: functional data indicating a loss-of-function mutation influencing the R122 trypsin cleavage site. AB - BACKGROUND: The understanding of genetic risk factors for chronic pancreatitis increased in the last decade with the discovery of mutations in the cationic trypsinogen gene (PRSS1). The first mutation was detected at the R122 autocleavage site of the protein (R122H) and subsequently two other mutations in this region, R122C and V123M, were described that resulted in a similar phenotype of hereditary pancreatitis. This study reports a novel A121T mutation within this region and characterises the resulting molecular properties at the autocleavage site. METHODS: Blood samples of a PRSS1 A121T carrier family were analysed for PRSS1 mutations using melting point curve analysis, restriction endonucleases and DNA sequencing. Conformation dependent properties of the mutated sequence were analysed by molecular modelling. The autodegradation kinetic of the mutated trypsin sequence was measured by a novel fluorescence resonance energy transfer (FRET) assay using designed 11 amino acid peptides from PRSS1 aa 118-aa 127 containing the trypsin cleavage site at aa 122 coupled to a Dabcyl/EDANS FRET system. The kinetic of tryptic peptide cleavage was measured in a fluorescence enzyme linked immunosorbent assay (ELISA) reader. RESULTS: DNA sequencing revealed a novel G to A transition at position 133279 of the published genomic sequence (#U66061 GenBank). The mutation results in an amino acid substitution of alanine by threonine at position 121 (A121T) of the cationic trypsinogen. Four additional mutation carriers could be identified among the relatives while only the first patient developed chronic pancreatitis. Molecular modelling of PRSS1 A121T revealed a change in the bond pattern between the R122 region and the calcium binding loop, whereas FRET assays showed an increased trypsin cleavage rate with a reaction kinetic elevated by more than 80%. CONCLUSION: The novel PRSS1 A121T mutation highlights the surface exposed region PRSS1 A121-R122-V123 as a hotspot for hereditary pancreatitis associated trypsinogen mutations. Molecular modelling and FRET assays provide evidence for an A121T mutation dependent increase in susceptibility to trypsin digestion at the R122 cleavage site suggesting an enhanced autodegradation and a loss-of-function at the autocleavage site. PMID- 18511572 TI - Separate endocytic pathways regulate IL-5 receptor internalization and signaling. AB - Eosinophils are critically dependent on IL-5 for their activation, differentiation, survival, and augmentation of cytotoxic activity. We previously showed that the cytoplasmic domain of the hematopoietic receptor, betac, which is shared by IL-5, IL-3, and GM-CSF, is directly ubiquitinated and degraded by the proteasomes in a JAK2-dependent manner. However, studies describing the spatial distribution, endocytic regulation, and trafficking of betac-sharing receptors in human eosinophils are currently lacking. Using deconvolution microscopy and biochemical methods, we clearly demonstrate that IL-5Rs reside in and are internalized by clathrin- and lipid raft-dependent endocytic pathways. Microscopy analyses in TF1 cells and human eosinophils revealed significant colocalization of betac, IL-5Ralpha, and Cy3-labeled IL-5 with transferrin- (clathrin) and cholera toxin-B- (lipid raft) positive vesicles. Moreover, whereas internalized IL-5Rs were detected in both clathrin- and lipid raft-positive vesicles, biochemical data revealed that tyrosine phosphorylated, ubiquitinated, and proteasome-degraded IL-5Rs partitioned to the soluble, nonraft fractions (clathrin-containing). Lastly, we show that optimal IL-5-induced signaling requires entry of activated IL-5Rs into the intracellular compartment, as coimmunoprecipitation of key signaling molecules with the IL-5R was completely blocked when either endocytic pathway was inhibited. These data provide the first evidence that IL-5Rs segregate and traffic into two distinct plasma membrane compartments, and they further establish that IL-5R endocytosis regulates signaling both positively and negatively. PMID- 18511573 TI - Novel PKC signaling is required for LPS-induced soluble Flt-1 expression in macrophages. AB - In vitro activation of macrophages by LPS induces rapid release of vascular endothelial growth factor (VEGF) and soluble fms-like tyrosine kinase-1 receptor (sFlt-1), which are thought to be the effectors to cause sepsis. However, the signal pathway that controls the VEGF and sFlt-1 expressions in LPS-activated macrophages remains unclear. In this study, we demonstrated that phosphorylation of protein kinase C (PKC)delta played a key role in the VEGF and sFlt-1 signaling pathway of LPS-activated macrophages. PKC is a family of serine-threonine kinases, which are classified into three major groups based on homology and cofactor requirements: conventional PKCs, novel PKCs, and atypical PKCs. In the murine RAW264.7 cells, as well as in primary human monocytes/macrophages, pretreatment with a general PKC inhibitor GF109203X or with a novel PKCdelta inhibitor rottlerin or overexpression of a kinase-inactive form of PKCdelta (K376R) eliminated LPS-induced sFlt-1 expression and augmented LPS-induced VEGF expression at the protein and the transcription levels. In contrast, Go6976, an inhibitor for the conventional PKCs, or myristoylated PKCzeta pseudosubstrate peptide, an inhibitor for the atypical PKCs, failed to exert the same effects. These data suggest that PKCdelta signaling is involved in LPS-induced sFlt-1 expression and serves as a negative mediator in LPS-induced VEGF expression in macrophages. A novel strategy controlling the LPS-induced PKC pathways, especially the PKCdelta isoform, may be developed based on this study. PMID- 18511574 TI - Conditioning response to granulocyte colony-stimulating factor via the dipeptidyl peptidase IV-adenosine deaminase complex. AB - G-CSF is routinely used to mobilize hematopoietic stem cells (HSCs) from bone marrow (BM) into peripheral blood before aphaeresis, but HSC harvesting can be suboptimal. On the other hand, transplanted HSCs sometimes fail to engraft a recipient BM microenvironment when G-CSF is used after transplantation, as pushing-CSF will push HSCs away from marrow. So, G-CSF action needs to be potentiated by other drugs. Marrow stromal cells establish a local CXCL12 concentration gradient that is the primary homing signal for HSCs. Pharmacological interventions that modify this gradient, therefore, have potential to help HSC mobilization (by decreasing CXCL12) and engraftment (by increasing CXCL12). CXCL12 inactivation is primarily mediated by dipeptidyl peptidase-IV. We review here the currently available drugs affecting this enzyme that could be used in the clinic to achieve phase-specific help for G-CSF. PMID- 18511576 TI - Commensal bacteria trigger a full dendritic cell maturation program that promotes the expansion of non-Tr1 suppressor T cells. AB - Dendritic cells (DCs) orchestrate the immune response establishing immunity versus tolerance. These two opposite functions may be dictated by DC maturation status with maturity linked to immunogenicity. DCs directly interact with trillions of noninvasive intestinal bacteria in vivo, a process that contributes to gut homeostasis. We here evaluated the maturation program elicited in human DCs by direct exposure to commensal-related bacteria (CB) in the absence of inflammatory signals. We showed that eight gram(+) and gram(-) CB strains up regulated costimulatory molecule expression in DCs and provoked a chemokine receptor switch similar to that activated by gram(+) pathogens. CB strains may be classified into three groups according to DC cytokine release: high IL-12 and low IL-10; low IL-12 and high IL-10; and low IL-12 and IL-10. All CB-treated DCs produced IL-1beta and IL-6 and almost no TGF-beta. Yet, CB instructed DCs to convert naive CD4+ T cells into hyporesponsive T cells that secreted low or no IFN-gamma, IL-10, and IL-17 and instead, displayed suppressor function. These data demonstrate that phenotypic DC maturation combined to an appropriate cytokine profile is insufficient to warrant Th1, IL-10-secreting T regulatory Type 1 (Tr1), or Th17 polarization. We propose that commensal flora and as such, probiotics manipulate DCs by a yet-unidentified pathway to enforce gut tolerance. PMID- 18511575 TI - The scavenger receptor SR-A I/II (CD204) signals via the receptor tyrosine kinase Mertk during apoptotic cell uptake by murine macrophages. AB - Apoptotic cells (AC) must be cleared by macrophages (Mo) to resolve inflammation effectively. Mertk and scavenger receptor A (SR-A) are two of many receptors involved in AC clearance. As SR-A lacks enzymatic activity or evident intracellular signaling motifs, yet seems to signal in some cell types, we hypothesized that SR-A signals via Mer receptor tyrosine kinase (Mertk), which contains a multisubstrate docking site. We induced apoptosis in murine thymocytes by dexamethasone and used Western blotting and immunoprecipitation to analyze the interaction of Mertk and SR-A in the J774A.1 (J774) murine Mo cell line and in peritoneal Mo of wild-type mice and SR-A-/- mice. Phagocytosis (but not adhesion) of AC by J774 was inhibited by anti-SR-A or function-blocking SR-A ligands. In resting J774, SR-A was associated minimally with unphosphorylated (monomeric) Mertk; exposure to AC induced a time-dependent increase in association of SR-A with Mertk in a direct or indirect manner. Anti-SR-A inhibited AC-induced phosphorylation of Mertk and of phospholipase Cgamma2, essential steps in AC ingestion. Relative to tissue Mo of wild-type mice, AC-induced Mertk phosphorylation was reduced and delayed in tissue Mo of SR-A-/- mice, as was in vitro AC ingestion at early time-points. Thus, during AC uptake by murine Mo, SR A is essential for optimal phosphorylation of Mertk and subsequent signaling required for AC ingestion. These data support the Mertk/SR-A complex as a potential target to manipulate AC clearance and hence, resolution of inflammation and infections. PMID- 18511577 TI - The measurement of psychopathy: dimensional and taxometric approaches. AB - This article extends the debate over personality disorders as dimensional or taxonic phenomena to the study of psychopathy and relates this issue to questions surrounding whether behaviors or personality traits best represent psychopathy. Proponents of dimensional measurements of psychopathy consider personality traits to be important constructs of psychopathy, whereas proponents of taxometric measurements consider behaviors to be important characteristics of psychopathy. After a brief introduction to the measurement of psychopathy, taxometric and dimensional measurement techniques are explained, their assumptions addressed, and their strengths and weaknesses discussed. Empirical evidence for each technique is then critiqued, and methodological problems are described. It is argued that methodological problems of existing studies largely preclude conclusions regarding whether psychopathy is dimensional or taxonic. Suggestions for future research are provided to address some of these methodological limitations. This review informs readers about each measurement approach and identifies problems regarding the dimensional or taxonic measurement of psychopathy. PMID- 18511578 TI - Where have all the "breakthroughs" gone? PMID- 18511579 TI - Economic grand rounds: a self-directed care model for mental health recovery. AB - Self-directed care programs give participants control over public funds to purchase services and supports for their own recovery. Data were examined for 106 individuals and showed that compared with the year before enrollment, in the year after enrollment, participants spent significantly less time in psychiatric inpatient and criminal justice settings and showed significantly better functioning. Of approximately $58,000 in direct expenditures by participants over 19 months of operation, 47% was spent on traditional psychiatric services, 13% on service substitutions for traditional care, 29% on tangible goods, 8% on uncovered medical care, and 3% on transportation. Early positive results of this pilot program support replication and evaluation elsewhere. PMID- 18511580 TI - Best practices: a program to support shared decision making in an outpatient psychiatric medication clinic. AB - This column presents preliminary findings of an intervention to support shared decision making in psychopharmacology consultation. The waiting area in an urban psychiatric medication clinic was transformed into a peer-run Decision Support Center featuring a user-friendly, Internet-based software program with which clients could create a one-page computer-generated report for use in the medication consultation. The Decision Support Center was used 662 times by 189 unique users from a young-adult and general adult case management team from October 2006 to September 2007. All clients had severe mental disorders. Only ten clients refused to use the intervention at some point during the pilot study. Focus groups with medical staff (N=4), clients (N=16), case managers (N=14), and peer-specialist staff (N=3) reported that the intervention helped to create efficiencies in the consultation and empower clients to become more involved in treatment-related decision making. A randomized controlled trial is currently in process. PMID- 18511581 TI - Personal accounts: a 35-year follow-up of completed psychotherapy. PMID- 18511582 TI - Trauma and PTSD among adolescents with severe emotional disorders involved in multiple service systems. AB - OBJECTIVE: This study examined the prevalence and correlates of posttraumatic stress disorder (PTSD) among adolescents with severe emotional disorders who were involved in multiple service systems. METHODS: Sixty-nine adolescents, ages 11 17, and their primary caregivers participated in a system-of-care project in three regions of New Hampshire and were interviewed to determine adolescent trauma exposure, prevalence of PTSD, treatment history, family background, behavioral and emotional problems, functioning, caregiver strain, and strengths and resilience. RESULTS: The rate of current PTSD was 28%, which was underdiagnosed in adolescents' medical records. PTSD was related to gender (42% for girls and 19% for boys; p=.03), history of sexual abuse (61% among youths with sexual abuse and 15% among youths without), chart diagnosis of depression (47% among youths with depression diagnoses and 16% among youths without), and treatment with multiple psychotropic medications (53% among youths prescribed two or more medications and 26% among those prescribed no medication or one medication). Adolescents with PTSD also were more likely to have run away, engaged in self-injurious and delinquent behavior, reported higher anxiety and depression, and functioned worse at school and home than those without PTSD. CONCLUSIONS: PTSD is a common but underdiagnosed disorder among adolescents with severe emotional and behavioral disorders who are involved in multiple service systems. Routine screening for trauma exposure and PTSD should be conducted with all adolescents receiving mental health services so that treatment can be provided to those with PTSD. PMID- 18511583 TI - Evaluation of universal screening for military-related sexual trauma. AB - OBJECTIVES: In response to growing concerns about sexual violence as an underrecognized traumatic consequence of military service, Veterans Health Administration policy requires universal screening for sexual trauma sustained during military service. This prospective study, the first to evaluate national efforts to screen for military sexual trauma, investigated whether sexual trauma screening is associated with increased utilization of mental health services. METHODS: This study examined data for all male (N=540,381) and female (N=33,259) veterans who had valid responses to screens for military sexual trauma in 2005. The use of mental health services during the three months after screening was examined for persons who screened positive for military sexual trauma and for those who screened negative. Findings were stratified by use of mental health services in the six months before the screening. RESULTS: Compared with negative screens, positive screens were associated with significantly increased rates of postscreen mental health treatment. A more than twofold increase was observed for patients without previous use of mental health treatment (women: relative risk [RR]=2.52, 95% confidence interval [CI]= 2.38-2.66; men: RR=2.47, 95% CI=2.34 2.61). In this group, the number of positive screens needed for one additional patient to access treatment was 5.5 for women and 7.2 for men. CONCLUSIONS: Our findings suggest that detection via screening is associated with increased rates of mental health treatment. An effective screening program that promotes detection of sexual trauma and access to mental health care can help to reduce the burden of psychiatric illness for those who have experienced military sexual trauma. PMID- 18511584 TI - Prevalence, diagnosis, and treatment of depression and generalized anxiety disorder in a diverse urban community. AB - OBJECTIVE: This study assessed the prevalence, diagnosis, and treatment of major depressive disorder and generalized anxiety disorder among New York City adults. METHODS: As part of the first community-specific Health and Nutrition Examination Survey in the United States, depression and anxiety were assessed in a representative sample of 1,817 noninstitutionalized adults in 2004. RESULTS: A total of 8% had major depressive disorder and 4% had generalized anxiety disorder. Respondents with depression were more likely to be formerly married, publicly insured, younger, and U.S. born. Only 55% of adults with depression were diagnosed, and 38% of those with depression or anxiety were in treatment; individuals with a diagnosis of depression were more likely to receive treatment than those without a diagnosis (61% versus 7%; p<.001). Immigrants with depression were 60% less likely to be diagnosed than their U.S.-born counterparts; immigrants arriving in this country ten or more years ago had slightly more anxiety than immigrants arriving less than ten years ago (3% versus 2%, not significant). Among respondents with anxiety, 23% reported disability compared with 15% of those with depression. Compared with adults with neither diagnosis, adults with depression or anxiety were twice as likely to smoke tobacco (p<.05), adults with depression were twice as likely to have diabetes (p<.01), and those with anxiety were twice as likely to have asthma (p<.01). CONCLUSIONS: Mental disorders are often disabling and inadequately diagnosed and treated. Foreign-born adults experience barriers to diagnosis and treatment despite having less depression; anxiety may increase with time since immigration. Increased awareness of and linkage to mental health services are needed, especially in larger, more diverse urban communities. PMID- 18511585 TI - The impact of immigration on psychiatric hospitalization in Illinois from 1993 to 2003. AB - OBJECTIVE: Illinois public hospitalizations over a ten-year period were studied to determine the impact of recent immigration. The study also explored clinical and demographic differences between immigrant groups and native-born Americans. METHODS: Information was collected from the state hospital Clinical Information System for 1993, 1998, and 2003. Variables included age, sex, race, marital status, education, diagnosis, length of stay, birthplace, citizenship, primary language, English proficiency, and availability of a Social Security number. Logistic multiple regression was used to analyze trends in the proportion of psychiatric admissions of foreign-born patients, with foreign born as the dependent variable and year as the independent variable. Chi square analysis was used for trends across time. RESULTS: In the hospitalized population, the proportion of immigrants was 7.3% in 1993, 10.9% in 1998, and 13.1% in 2003. With covariates adjusted for, the average increase of 8.0% per year in the odds of being foreign born was statistically significant (odds ratio=1.08, 95% confidence interval=1.06-1.10). Nevertheless, the proportion of foreign-born hospital admissions, including Asian and Mexican immigrants, was below their population ratio in Illinois. Mexican-origin immigrants constituted the largest group of admissions and were younger, less educated, had poorer English skills, and were more likely to be undocumented than other immigrants. CONCLUSIONS: The percentage of foreign-born patients admitted was lower than their percentage in the overall population. In previous immigration waves, immigrants were hospitalized at disproportionately higher rates than nonimmigrants. The gap is slowly narrowing as new admissions are increasingly likely to be foreign born, suggesting that public psychiatric hospitals should prepare for these changing populations. PMID- 18511586 TI - Unmet needs of families of adults with mental illness and preferences regarding family services. AB - OBJECTIVE: This study used a survey to assess the information and educational needs of family members of adults with mental illness and their preferences regarding how to address those needs. METHODS: Recruitment was attempted through two sources: local mental health treatment facilities and the Maryland chapter of the National Alliance on Mental Illness (NAMI). Inadequate contact information and low response rate produced only 16 responses from family members of consumers recruited through local mental health facilities. Thus results are reported for a family needs assessment survey mailed to NAMI members (308 of 962 possible responses). Bivariate and multivariate analyses were used to summarize relationships between characteristics of the family member, characteristics of the ill relative, experience of stigma by the family member, and information needs of the family members. RESULTS: On average, family members reported a substantial number of unmet needs (mean+/-SD of 7.09+/-4.71 needs; possible number of needs ranges from 0 to 16), often despite prior receipt of information. Family members' experiences of stigma and having an ill relative with a more recently occurring condition (for example, a younger relative or a shorter length of illness) or with a disabling condition (for example, recent hospitalization) were significantly associated with a greater number of unmet needs. Family members preferred that a mental health provider (63%) address their needs on an as-needed basis (58%). CONCLUSIONS: The needs and preferences of family members of adults with mental illness are diverse and varied. Consequently, these families may benefit from ongoing provision of information and support tailored to meet the families' individual needs. Continued efforts should be made to understand and address consumer and family needs, potential barriers to participation in family services, and the relationship between stigma and family need. PMID- 18511587 TI - Gender differences in health-related quality of life for veterans with serious mental illness. AB - OBJECTIVE: This study assessed gender differences in health-related quality of life (HRQOL) in a national sample of veterans with serious mental illness. METHODS: Data were analyzed from the Large Health Survey of Veterans, which was mailed to a national random sample of veterans in 1999. The linear and logistic multiple regression analyses included 18,017 veterans with schizophrenia, schizoaffective disorder, or bipolar disorder who completed the survey. HRQOL was measured by using the various subscales of the 36-Item Short Form of the Medical Outcomes Study (MOS SF-36) (mental component summary, physical component summary, and activities of daily living) and by questions assessing self-perceptions of health status. RESULTS: The sample was 7.3% female, 75.7% white, and 83.8% unemployed. Mean+/-SD age was 54.3+/-12.2 years. After the analysis adjusted for sociodemographic characteristics, health status, and other variables, compared with male veterans, female veterans with serious mental illness had lower scores on the SF-36 physical component summary (indicating worse symptoms), were more likely to report that they were limited "a lot" in activities of daily living, and had more pain. However, female respondents were more likely to have a positive outlook on their health. CONCLUSIONS: Among veterans who received a diagnosis of serious mental illness from providers of the Department of Veterans Affairs, women reported substantially poorer HRQOL than men across several domains but women reported better self-perceived health. Attention to the particular needs of female veterans with serious mental illness is imperative as the numbers of female veterans continue to increase. PMID- 18511588 TI - Perceptions about competing psychosocial problems and treatment priorities among older adults with depression. AB - OBJECTIVE: Depression often co-occurs with other conditions that may pose competing demands to depression care, particularly in later life. This study examined older adults' perceptions of depression among co-occurring social, medical, and functional problems and compared the priority of depression with that of other problems. METHODS: The study's purposeful sample comprised 49 adults age 60 or older with a history of depression and in publicly funded community long-term care. Four-part, mixed-methods interviews sought to capture participants' perceptions of life problems as well as the priority they placed on depression. Methods included standardized depression screening, semistructured qualitative interviews, listing of problems, and qualitative and quantitative analysis of problem rankings. RESULTS: Most participants identified health, functional, and psychosocial problems co-occurring with depressive symptoms. Depression was ranked low among the co-occurring conditions; 6% ranked depression as the most important of their problems, whereas 45% ranked it last. Relative rank scores for problems were remarkably similar, with the notable exception of depression, which was ranked lowest of all problems. Participants did not see depression as a high priority compared with co-occurring problems, particularly psychosocial ones. CONCLUSIONS: Effective and durable improvements to mental health care must be shaped by an understanding of client perceptions and priorities. Motivational interviewing, health education, and assessment of treatment priorities may be necessary in helping older adults value and accept depression care. Nonspecialty settings of care may effectively link depression treatment to other services, thereby increasing receptivity to mental health services. PMID- 18511589 TI - Review of completed suicides in the California Department of Corrections and Rehabilitation, 1999 to 2004. AB - OBJECTIVE: The purpose of this extended review is to assist health care managers, clinicians, prison administrators, and custody staff in identifying and responding effectively to prisoners who present a substantial risk of suicide in the foreseeable future. METHODS: The California Department of Corrections and Rehabilitation (CDCR) is the largest state-operated prison system in the country, with a census range of 155,365 to 163,346 prisoners between 1999 and 2004. The authors conducted a review of all 154 suicides that occurred in CDCR during this period and examined several factors related to the suicide, including demographic characteristics of the inmate, health care information, suicide method, custody information, and emergency response. RESULTS: The analysis of trends in this six year review reveals that prisoners who completed suicide were similar to those who took their lives in the community in age distribution and mental health factors. The analysis also found that this group of prisoners who committed suicide had other characteristics or commonalities related specifically to their incarceration. In this review 60% of the suicides were judged to have been foreseeable, preventable, or both. CONCLUSIONS: Although suicide is not predictable, the terms "foreseeable" and "preventable" are used to indicate cases in which the risk of suicide was elevated or events occurred that should have triggered clinical or custodial reactions that would have reduced the likelihood of completed suicide. This review provides clues to recognize inmates at elevated risk and identifies some of the health care practices and conditions of confinement to consider for provision of an adequate suicide prevention program. PMID- 18511590 TI - Use of an expanded version of the DSM-IV outline for cultural formulation on a cultural consultation service. AB - OBJECTIVES: This study assessed cultural consultants' impression of the utility of an expanded version of the DSM-IV Outline for Cultural Formulation in cultural psychiatric consultation and identified ways to improve the usefulness of the cultural formulation. METHODS: A structured interview and questionnaire on the use of the cultural formulation was administered to 60 consultants working for an outpatient Cultural Consultation Service (CCS). RESULTS: Most consultants (93%) found the cultural formulation to be moderately to very useful. More than half (57%) had little or no familiarity with the cultural formulation before working with the CCS. The main suggestions for improvement of the cultural formulation were to expand sections on migration experience and include sections on religious and spiritual practice. CONCLUSIONS: Although many consultants had little previous familiarity with the cultural formulation, most found it useful in organizing their assessment and preparing consultation reports. The cultural formulation is a useful tool for nonmedical consultants and culture brokers, as well as for clinicians. PMID- 18511591 TI - American Association of Community Psychiatrists' views on general features of DSM IV. AB - OBJECTIVE: The authors report on a survey of the American Association of Community Psychiatrists (AACP) about improving DSM-IV. METHODS: An anonymous survey was sent to 600 psychiatrists of the AACP via Survey Monkey technology. RESULTS: Respondents (N=152) answered questionnaires regarding the general features of DSM-IV. Reliable interclinician communication was valued most highly. A majority of respondents (92%) reported using axis 1, 75% used axes 2 and 3, and approximately 50% used axes 4 and 5. AACP members were less keen on using the tool to inform patient management planning. Least valued were usefulness for a national statistical base or to indicate prognosis. CONCLUSIONS: AACP respondents' views suggest modification to the DSM system to improve clinical utility. Most favored fewer than 100 diagnostic categories. Many were concerned about the current systems' cultural sensitivity and accessibility to patients. These considerations should guide DSM-V deliberations. PMID- 18511592 TI - Assessing psychosocial stressors among Hispanic outpatients: does clinician ethnicity matter? AB - OBJECTIVE: Psychosocial and environmental stressors are a well-documented factor in the etiology, progression, and maintenance of psychiatric disorders. Clear guidelines on identifying them are lacking. When the patient and provider are of different cultures, the clinician may not properly understand and identify stressors. This study explored clinician ethnicity and identification of stressors. METHODS: A total of 88 adult Hispanic outpatients in a community clinic were separately evaluated by pairs of clinicians (Hispanic and non Hispanic) drawn from a pool of 47, as part of a larger study. Axis IV data are reported here. RESULTS: Clinicians identified few psychosocial stressors. Non Hispanic clinicians identified significantly more problems related to the primary support group and educational problems than Hispanic clinicians. CONCLUSIONS: Clinician ethnicity played a role in identification of psychosocial and environmental problems. Because stressors often affect the presenting problem and course of treatment, failure to properly identify and address them in treatment may influence service outcomes. PMID- 18511593 TI - Roundtable sessions of a Children's Hospital child protection team. PMID- 18511594 TI - "Meet the expert": cognitive problem-solving cancer support group. PMID- 18511595 TI - Problem gamblers' interest in self-help services. PMID- 18511596 TI - Policy implications of CATIE. PMID- 18511597 TI - Diverse adult stem cells share specific higher-order patterns of gene expression. AB - Adult tissue stem cells (SCs) share functional properties regardless of their tissue of residence. It had been thought that SCs might also share expression of certain "stemness" genes, although early investigations for such genes were unsuccessful. Here, we show that SCs from diverse tissues do preferentially express certain types of genes and that SCs resemble other SCs in terms of global gene expression more than they resemble the differentiated cells (DCs) of the tissues that they supply. Genes associated with nuclear function and RNA binding were over-represented in SCs. In contrast, DCs from diverse tissues shared enrichment in genes associated with extracellular space, signal transduction, and the plasma membrane. Further analysis showed that transit-amplifying cells could be distinguished from both SCs and DCs by heightened expression of cell division and DNA repair genes and decreased expression of apoptosis-related genes. This transit-amplifying cell-specific signature was confirmed by de novo generation of a global expression profile of a cell population highly enriched for transit amplifying cells: colonic crypt-base columnar cells responding to mucosal injury. Thus, progenitor cells preferentially express intracellular or biosynthetic genes, and differentiation correlates with increased expression of genes for interacting with other cells or the microenvironment. The higher-order, Gene Ontology term-based analysis we use to distinguish SC- and DC-associated gene expression patterns can also be used to identify intermediate differentiation states (e.g., that of transit-amplifying cells) and, potentially, any biological state that is reflected in changes in global gene expression patterns. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511598 TI - Hypermethylation of CXCR4 promoter in CD34+ cells from patients with primary myelofibrosis. AB - Constitutive mobilization of CD34(+) cells in patients with primary myelofibrosis (PMF) has been attributed to proteolytic disruption of the CXCR4/SDF-1 axis and reduced CXCR4 expression. We document here that the number of circulating CD34(+)/CXCR4(+) cells in PMF patients, as well as the cellular CXCR4 expression, was directly related to CXCR4 mRNA level and that reduced CXCR4 mRNA level was not due to SDF-1-induced downregulation. To address whether epigenetic regulation contributes to defective CXCR4 expression, we studied the methylation status of the CXCR4 promoter using methylation-specific polymerase chain reaction and methylation-specific sequencing in the JAK2V617F-positive HEL cell line and in CD34(+) cells. We found that CD34(+) cells from PMF patients, unlike those from normal subjects, presented hypermethylation of CXCR4 promoter CpG island 1. Following incubation with the demethylating agent 5-Aza-2'-deoxycytidine (5 AzaD), the percentage of PMF CD34(+) cells expressing CXCR4 increased 3-10 times, whereas CXCR4 mRNA level increased approximately 4 times. 5-AzaD-treated PMF CD34(+) cells displayed almost complete reversal of CpG1 island 1 hypermethylation and showed enhanced migration in vitro in response to SDF-1. These data point to abnormal methylation of the CXCR4 promoter as a mechanism contributing to constitutive migration of CD34(+) cells in PMF. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511599 TI - Improved efficiency and pace of generating induced pluripotent stem cells from human adult and fetal fibroblasts. AB - It was reported recently that human fibroblasts can be reprogrammed into a pluripotent state that resembles that of human embryonic stem (hES) cells. This was achieved by ectopic expression of four genes followed by culture on mouse embryonic fibroblast (MEF) feeders under a condition favoring hES cell growth. However, the efficiency of generating human induced pluripotent stem (iPS) cells is low, especially for postnatal human fibroblasts. We started supplementing with an additional gene or bioactive molecules to increase the efficiency of generating iPS cells from human adult as well as fetal fibroblasts. We report here that adding SV40 large T antigen (T) to either set of the four reprogramming genes previously used enhanced the efficiency by 23-70-fold from both human adult and fetal fibroblasts. Discernible hES-like colonies also emerged 1-2 weeks earlier if T was added. With the improved efficiency, we succeeded in replacing MEFs with immortalized human feeder cells that we previously established for optimal hES cell growth. We further characterized individually picked hES-like colonies after expansion (up to 24 passages). The majority of them expressed various undifferentiated hES markers. Some but not all the hES-like clones can be induced to differentiate into the derivatives of the three embryonic germ layers in both teratoma formation and embryoid body (EB) formation assays. These pluripotent clones also differentiated into trophoblasts after EB formation or bone morphogenetic protein 4 induction as classic hES cells. Using this improved approach, we also generated hES-like cells from homozygous fibroblasts containing the sickle cell anemia mutation Hemoglobin Sickle. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511600 TI - Fate-mapping evidence that hepatic stellate cells are epithelial progenitors in adult mouse livers. AB - Liver injury activates quiescent hepatic stellate cells (Q-HSC) to proliferative myofibroblasts. Accumulation of myofibroblastic hepatic stellate cells (MF-HSC) sometimes causes cirrhosis and liver failure. However, MF-HSC also promote liver regeneration by producing growth factors for oval cells, bipotent progenitors of hepatocytes and cholangiocytes. Genes that are expressed by primary hepatic stellate cell (HSC) isolates overlap those expressed by oval cells, and hepatocytic and ductular cells emerge when HSC are cultured under certain conditions. We evaluated the hypothesis that HSC are a type of oval cell and, thus, capable of generating hepatocytes to regenerate injured livers. Because Q HSC express glial fibrillary acidic protein (GFAP), we crossed mice in which GFAP promoter elements regulated Cre-recombinase with ROSA-loxP-stop-loxP-green fluorescent protein (GFP) mice to generate GFAP-Cre/GFP double-transgenic mice. These mice were fed methionine choline-deficient, ethionine-supplemented diets to activate and expand HSC and oval cell populations. GFP(+) progeny of GFAP expressing precursors were characterized by immunohistochemistry. Basal expression of mesenchymal markers was negligible in GFAP(+)Q-HSC. When activated by liver injury or culture, HSC downregulated expression of GFAP but remained GFP(+); they became highly proliferative and began to coexpress markers of mesenchyme and oval cells. These transitional cells disappeared as GFP-expressing hepatocytes emerged, began to express albumin, and eventually repopulated large areas of the hepatic parenchyma. Ductular cells also expressed GFAP and GFP, but their proliferative activity did not increase in this model. These findings suggest that HSC are a type of oval cell that transitions through a mesenchymal phase before differentiating into hepatocytes during liver regeneration. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511601 TI - Hypoxic preconditioning results in increased motility and improved therapeutic potential of human mesenchymal stem cells. AB - Mesenchymal stem cells (MSC) are adult multipotent cells found in bone marrow, adipose tissue, and other adult tissues. MSC have been shown to improve regeneration of injured tissues in vivo, but the mechanisms remain unclear. Typically, MSC are cultured under ambient, or normoxic, conditions (21% oxygen). However, the physiological niches for MSC in the bone marrow and other sites have much lower oxygen tension. When used as a therapeutic tool to repair tissue injuries, MSC cultured in standard conditions must adapt from 21% oxygen in culture to less than 1% oxygen in the ischemic tissue. We therefore examined the effects of preculturing human bone marrow-derived MSC in hypoxic conditions (1% 3% oxygen) to elucidate the best conditions that enhance their tissue regenerative potential. We demonstrated that MSC cultured in hypoxia activate the Akt signaling pathway while maintaining their viability and cell cycle rates. We also showed that MSC cultured in hypoxia induced expression of cMet, the major receptor for hepatocyte growth factor (HGF), and enhanced cMet signaling. MSC cultured in hypoxic conditions increased their migration rates. Since migration and HGF responsiveness are thought to be key mediators of MSC recruitment and/or activation in vivo, we next examined the tissue regenerative potential of MSC cultured under hypoxic conditions, using a murine hind limb ischemia model. We showed that local expression of HGF is increased in ischemic muscle in this model. Intra-arterial injection of MSC cultured in either normoxic or hypoxic conditions 24 hours after surgical induction of hind limb ischemia enhanced revascularization compared with saline controls. However, restoration of blood flow was observed significantly earlier in mice that had been injected with hypoxic preconditioned MSC. Collectively, these data suggest that preculturing MSC under hypoxic conditions prior to transplantation improves their tissue regenerative potential. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511602 TI - Characterization of a novel ubiquitin-conjugating enzyme that regulates beta1,4 galactosyltransferase-1 in embryonic stem cells. AB - In this study we identified a novel galactosyltransferase 1-associating protein (GTAP) by cDNA cloning from a murine embryonic cDNA library using the two-hybrid yeast system. GTAP is expressed in early embryonic tissues, as well as in adult tissues with active cell turnover, and belongs to the class III ubiquitin conjugating (E2) enzyme family. Its COOH-terminal domain contains a consensus sequence for ubiquitin binding shared by all the ubiquitin-conjugating enzymes, whereas its NH(2)-terminal domain appears critical for the binding and internalization of cell surface galactosyltransferase 1 (GalT1) in embryonic stem cells through a monensin- and MG132-dependent pathway. We have found that GTAP regulates GalT1-associated, laminin-dependent embryonic cell adhesion and the formation of embryoid bodies. Thus, GTAP functions as an evolutionarily conserved E2 enzyme, which may participate in intercellular adhesion and embryonic development. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511603 TI - Hypoxia causes downregulation of mismatch repair system and genomic instability in stem cells. AB - The DNA mismatch repair (MMR) system maintains genomic integrity by correcting replication errors: its malfunction causes genomic instability in several tumor types. Hypoxia-inducible factor-1alpha (HIF1alpha), the major regulator of the processes that occur in hypoxia and certain epigenetic events downregulate the expression of MMR genes in cancer cells. However, there is a lack of information regarding MMR regulation and the genetic stability of stem cells under hypoxic conditions. The expression of the MMR system is downregulated in murine and human stem cells cultured in hypoxia, which correlates with lower DNA repair activity in neural stem cells. We observed, through the use of short hairpin loop RNAi expression constructs, that HIF1alpha positively regulated MLH1 and MSH6 when the C17.2 neural stem cells were exposed to short-term hypoxia. However, in prolonged exposure to oxygen depletion, the reduced transcriptional activation of MMR genes was directed by specific epigenetic events. Chromatin immunoprecipitation experiments showed a hypoacetylated/hypermethylated histone H3 and lower SP1 binding within MLH1 and MSH6 adjacent promoter regions. Treatment with the histone deacetylase inhibitor trichostatin A increased histone H3 acetylation and SP1 occupancy and enhanced MMR expression. Sequencing of microsatellite markers revealed genomic instability in the murine and human stem cells grown under hypoxia. Thus, the present article reports, for the first time in the stem cell field, experimental data that indicate that hypoxic niches are an environment in which stem cells might undergo genomic instability, which could lie at the origin of subpopulations with cancer stem cell properties. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511604 TI - Evidence that very small embryonic-like stem cells are mobilized into peripheral blood. AB - Recently, we identified in murine adult tissues, including bone marrow, a population of very small embryonic-like (VSEL) stem cells. Here, we provide further evidence that under steady-state conditions these cells circulate at very low levels in peripheral blood (PB) ( approximately 100-200 cells/ml) and could be additionally mobilized during pharmacological granulocyte-colony-stimulating factor-induced or stress-related mobilization, as demonstrated in a model of toxic liver or skeletal muscle damage induced by injection of carbon tetrachloride or cardiotoxin, respectively. The number of circulating VSEL stem cells under steady-state conditions in PB of 2-month-old animals was five times higher than that in 1-year-old mice. In conclusion, this study supports a hypothesis that VSEL stem cells are a mobile pool of primitive stem cells that could be released from the stem cell niches into PB. Further studies are needed, however, to see whether the level of these cells circulating in PB could become a prognostic indicator to assess the regenerative potential of an adult organism and/or clinical outcome from an injury. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18511605 TI - Effect of amlodipine on cardiovascular events in hypertensive haemodialysis patients. AB - BACKGROUND: Hypertensive haemodialysis patients may be at a high risk for cardiovascular events. This study was undertaken to ascertain whether the calcium channel blocker amlodipine reduces mortality and cardiovascular events in these high-risk patients. METHODS: We evaluated the effects of amlodipine on cardiovascular events in 251 hypertensive haemodialysis patients in an investigator-designed, prospective, randomized, double-blind, placebo-controlled, multicenter trial. One hundred and twenty-three patients were randomly assigned to amlodipine (10 mg once daily) and 128 to placebo. The primary endpoint was mortality from any cause. The secondary endpoint was a composite variable consisting of mortality from any cause or cardiovascular event. Analysis was by intention-to-treat. The trial was registered with ClinicalTrials.gov (number NCT00124969). RESULTS: The median age of patients was 61 years (25% percentile - 75% percentile, 47-69), and the median follow-up was 19 months (8-30). Fifteen (12%) of the 123 patients assigned to amlodipine and 22 (17%) of the 128 patients assigned to placebo had a primary endpoint [hazard ratio 0.65 (95% CI 0.34-1.23); P = 0.19]. Nineteen (15%) of the 123 haemodialysis patients assigned to amlodipine and 32 (25%) of the 128 haemodialysis patients assigned to placebo reached the secondary composite endpoint [hazard ratio 0.53 (95% CI 0.31-0.93); P = 0.03]. CONCLUSION: Amlodipine safely reduces systolic blood pressure and it may have a beneficial effect on cardiovascular outcomes in hypertensive haemodialysis patients. PMID- 18511606 TI - Vascular access use and outcomes: an international perspective from the Dialysis Outcomes and Practice Patterns Study. AB - BACKGROUND: A well-functioning vascular access (VA) is essential to efficient dialysis therapy. Guidelines have been implemented improving care, yet access use varies widely across countries and VA complications remain a problem. This study took advantage of the unique opportunity to utilize data from the Dialysis Outcomes and Practice Patterns Study (DOPPS) to examine international trends in VA use and trends in patient characteristics and practices associated with VA use from 1996 to 2007. DOPPS is a prospective, observational study of haemodialysis (HD) practices and patient outcomes at >300 HD units from 12 countries and has collected data thus far from >35,000 randomly selected patients. METHODS: VA data were collected for each patient at study entry (1996-2007). Practice pattern data from the facility medical director, nurse manager and VA surgeon were also analysed. RESULTS: Since 2005, a native arteriovenous fistula (AVF) was used by 67-91% of prevalent patients in Japan, Italy, Germany, France, Spain, the UK, Australia and New Zealand, and 50-59% in Belgium, Sweden and Canada. From 1996 to 2007, AVF use rose from 24% to 47% in the USA but declined in Italy, Germany and Spain. Moreover, graft use fell by 50% in the USA from 58% use in 1996 to 28% by 2007. Across three phases of data collection, patients consistently were less likely to use an AVF versus other VA types if female, of older age, having greater body mass index, diabetes, peripheral vascular disease or recurrent cellulitis/gangrene. In addition, countries with a greater prevalence of diabetes in HD patients had a significantly lower percentage of patients using an AVF. Despite poorer outcomes for central vein catheters, catheter use rose 1.5- to 3 fold among prevalent patients in many countries from 1996 to 2007, even among non diabetic patients 18-70 years old. Furthermore, 58-73% of patients new to end stage renal disease (ESRD) used a catheter for the initiation of HD in five countries despite 60-79% of patients having been seen by a nephrologist >4 months prior to ESRD. Patients were significantly (P < 0.05) less likely to start dialysis with a permanent VA if treated in a faciity that (1) had a longer time from referral to access surgery evaluation or from evaluation to access creation and (2) had longer time from access creation until first AVF cannulation. The median time from referral until access creation varied from 5-6 days in Italy, Japan and Germany to 40-43 days in the UK and Canada. Compared to patients using an AVF, patients with a catheter displayed significantly lower mean Kt/V levels. CONCLUSIONS: Most countries meet the contemporary National Kidney Foundation's Kidney Disease Outcomes Quality Initiative goal for AVF use; however, there is still a wide variation in VA preference. Delays between the creation and cannulation must be improved to enhance the chances of a future permanent VA. Native arteriovenous fistula is the VA of choice ensuring dialysis adequacy and better patient outcomes. Graft is, however, a better alternative than catheter for patients where the creation of an attempted AVF failed or could not be created for different reasons. PMID- 18511607 TI - Bacterial DNA and endothelial damage in haemodialysis patients. AB - BACKGROUND: An increased percentage of CD14+CD16+ activated monocytes have been reported in peripheral blood from haemodialysis patients. The aim of this study is to investigate if a mild stimulus such as bacterial DNA (CpG-ODNs) contamination may induce an inflammatory response in CD14+CD16+ monocytes from haemodialysis patients and to value the biological consequences of this inflammatory response on endothelial cell damage. METHODS: Circulating mononuclear cells from 20 haemodialysis patients and 15 healthy subjects were studied. CD14+CD16+ and the toll-like receptor 9 (TLR-9) expression were assessed by flow cytometry. Cell culture inserts were used to evaluate the effect of CD14+CD16+ and CpG-ODNs on endothelial cell apoptosis (measured by Tunnel). Intracellular cytokines were measured by Cytometric methods. NF-kappaB, p38 MAPK, c-Jun PI3K and MEK1/2 activity were modified by specific peptides. RESULTS: At baseline, CD14+CD16+ have an increased expression of cytoquines and TLR-9. CpG ODNs caused the production and release of cytoquines in CD14+CD16+, but not in CD14++ monocytes. This inflammatory response was mediated by intracellular signalling dependent on NF-kappaB, p38 MARK or c-Jun PI3K but not by MEK1/2 activation. The results of the present study also demonstrate that the inflammatory response induced by the stimulation of CD14+CD16+ by CpG DNA resulted in endothelial cell apoptosis. CONCLUSIONS: The results of the present study demonstrate that in haemodialysis patients there is a subpopulation of pre activated monocytes that can be stimulated by contaminant bacterial DNA. These activated cells produce and release inflammatory factors that may cause endothelial injury. PMID- 18511608 TI - Adding access blood flow surveillance to clinical monitoring reduces thrombosis rates and costs, and improves fistula patency in the short term: a controlled cohort study. AB - BACKGROUND: Access blood flow (Qa) measurement is the recommended method for fistula (AVF) surveillance for stenosis, but whether it may be beneficial and cost-effective is controversial. METHODS: We conducted a 5-year controlled cohort study to evaluate whether adding Qa surveillance to unsystematic clinical monitoring (combined with elective stenosis repair) reduces thrombosis and access loss rates, and costs in mature AVFs. We prospectively collected data in 159 haemodialysis patients with mature AVFs, 97 followed by unsystematic clinical monitoring (Control) and 62 by adding Qa surveillance to monitoring (Flow). Indications for imaging and stenosis repair were clinically evident access dysfunction in both groups and a Qa < 750 ml/min or dropping by >20% in Flow. RESULTS: Adding Qa surveillance prompted an increase in access imaging (HR 2.96, 95% CI 1.79-4.91, P < 0.001), stenosis detection (HR 2.55, 95% CI 1.48-4.42, P = 0.001) and elective repair (HR 2.26, 95% CI 1.16-4.43, P = 0.017), and a reduction in thromboses (HR 0.27, 95% CI 0.09-0.79, P = 0.017), central venous catheter placements (HR 0.14, 95% CI 0.03-0.42, P = 0.010) and access losses (HR 0.35, 95% CI 0.11-1.09, P = 0.071). In the Kaplan-Meier analysis, adding Qa surveillance only extended short-term cumulative patency (P = 0.037 in the Breslow test). Mean access-related costs were 1213 Euro/AVF-year in Control and 743 in Flow (P < 0.001). CONCLUSIONS: Our controlled cohort study shows that adding Qa surveillance to monitoring in mature AVFs is associated with a better detection and elective treatment of stenosis, and lower thrombosis rates and access-related costs, although the cumulative access patency was only extended in the first 3 years after fistula maturation. We are aware of the limitations of our study (non-randomization and the possible centre effect) and that further, better-designed trials are needed to arrive at a definitive answer concerning the role of Qa surveillance for fistulae. PMID- 18511609 TI - Acute, severe and anaphylactoid reactions are very rare with low-molecular-weight iron dextran, CosmoFer. PMID- 18511610 TI - Phase IV research: innovation in need of ethics. PMID- 18511611 TI - The "future like ours" argument and human embryonic stem cell research. AB - The most closely argued and widely discussed case against abortion in the philosophical literature today is Don Marquis's "future like ours" argument. The argument moves from an analysis of why there is a serious presumption against killing someone "like us" to the conclusion that most abortions are seriously wrong for the same reason: they deprive "an individual" of a future of valuable experiences and activities, a "future like ours". Julian Savulescu has objected that "preventing" such a future could not be as seriously presumptively wrong as Marquis contends for if it were, even contraception and failure to engage in reproductive cloning would be seriously presumptively wrong. Savulescu maintains that there is only a modest presumption against preventing a "future of value like ours" and that in the case of human embryonic stem cell research, it is clearly outweighed by "the enormous potential to save people's lives and to improve their quality of life". Marquis defends his strong anti-abortion stance against Savulescu's "contraception" and "failure to clone" objections but surprisingly says nothing about the implications of the "future like ours" argument for the controversy surrounding human embryonic stem cell research. I argue that key features of Marquis's response actually support the view that embryos used in stem cell research are not included within the protective scope of the "future like ours" argument. It is significant that the most philosophically rigorous anti-abortion case thus far presented does not entail that human embryonic stem cell research is even presumptively wrong. PMID- 18511612 TI - Abortion and human nature. PMID- 18511613 TI - How do elderly spouse care givers of people with Alzheimer disease experience the disclosure of dementia diagnosis and subsequent care? AB - OBJECTIVES: To examine the experiences of spousal care givers of Alzheimer patients to disclosure of dementia diagnosis and subsequent care. METHODS: A random sample of 1943 spousal care givers of people receiving medication for Alzheimer disease (AD) was sent a cross-sectional postal survey about their opinions on the disclosure of dementia and follow-up care. A smaller qualitative study (n = 63) included open-ended questions concerning their experiences of the same topics. RESULTS: The response rate for the survey was 77%. Of the respondents, 1214 of 1434 acknowledged themselves as their spouse's care giver. The mean age of the care givers was 78.2 years, and that of demented spouses, 80.5 years. Of the care givers, 63% were women. The couples had long-lasting marriages (mean 52 years). Of the care givers, 93% reported that dementia had been disclosed openly to their spouse; 97% also preferred that physicians openly inform the patients of the dementia diagnosis, although 55% of their spouses with AD had developed depressive symptoms after the disclosure. Of the care givers, 71% felt they had received sufficient information about dementia. However, only 50% estimated that their spouses' follow-up care had been well organised. The responses in the qualitative study indicated that many care givers felt grief and anxiety. They also expressed feelings of loneliness and uncertainty about how to deal with follow-up care for dementia. CONCLUSIONS: Elderly spousal care givers were quite satisfied with the information given them about dementia. However, the support with regard to the follow-up care of care-giving families failed to meet their needs adequately. PMID- 18511614 TI - Nurses' attitudes towards artificial food or fluid administration in patients with dementia and in terminally ill patients: a review of the literature. AB - OBJECTIVE: Although nurses have an important role in the care process surrounding artificial food or fluid administration in patients with dementia or in terminally ill patients, little is known about their attitudes towards this issue. The purpose of this study was to thoroughly examine nurses' attitudes by means of a literature review. METHOD: An extensive systematic search of the electronic databases PubMed, Cinahl, PsycINFO, The Cochrane Library, FRANCIS, Philosopher's Index and Social Sciences Citation Index was conducted to identify pertinent articles published from January 1990 to January 2007. FINDINGS: Nurses' arguments for or against could be categorised as ethical-legal, clinical or social-professional. The most important arguments explicitly for artificial food and fluid administration in patients with dementia or in terminally ill patients were sanctity of life, considering artificial food and fluid administration as basic nursing care, and giving reliable nutrition, hydration or medication. An explicit counter-argument was the high cost of treatment. Arguments used by opponents and proponents were quality of life and dignified death. The arguments were not strikingly different for the two patient populations. It turned out that the nurses' ethical arguments remarkably reflected the current ethical debate. But some of their clinical presuppositions contradicted current clinical evidence. CONCLUSION: The interaction between clinical facts and ethical reflections makes the findings of this review extremely relevant for clinical ethics. A large need exists to clearly communicate to nurses the latest clinical evidence and the main results of ongoing ethical debates. PMID- 18511615 TI - Medically valid religious beliefs. AB - Patient requests for "inappropriate" medical treatment (violations of the standard of care) based on religious beliefs should have special standing. Nevertheless, not all such requests should be honored, because some are morally disturbing. The trouble lies in deciding which ones count. This paper proposes criteria that would qualify a religious belief as medically valid to help physicians decide which requests to respect. The four conditions suggested are that the belief (1) is shared by a community, (2) is deeply held, (3) would pass the test of a religious interpreter and (4) does not harm others. PMID- 18511616 TI - Ethical decision-making, passivity and pharmacy. AB - BACKGROUND: Increasing interest in empirical ethics has enhanced understanding of healthcare professionals' ethical problems and attendant decision-making. A four stage decision-making model involving ethical attention, reasoning, intention and action offers further insights into how more than reasoning alone may contribute to decision-making. AIMS: To explore how the four-stage model can increase understanding of decision-making in healthcare and describe the decision-making of an under-researched professional group. METHODS: 23 purposively sampled UK community pharmacists were asked, in semi-structured interviews, to describe ethical problems in their work and how they were resolved. Framework analysis of transcribed interviews utilised the four decision-making stages, together with constant comparative methods and deviant-case analysis. RESULTS: Pharmacists were often inattentive and constructed problems in legal terms. Ethical reasoning was limited, but examples of appeals to consequences, the golden rule, religious faith and common-sense experience emerged. Ethical intention was compromised by frequent concern about legal prosecution. Ethical inaction was common, typified by pharmacists' failure to report healthcare professionals' bad practices, and ethical passivity emerged to describe these negative examples of the four decision-making stages. Pharmacists occasionally described more ethically active decision-making, but this often involved ethical uncertainty. DISCUSSION: The four decision-making stages are a useful tool in considering how healthcare professionals try to resolve ethical problems in practice. They reveal processes often ignored in normative theories, and their recognition and the emergence of ethical passivity indicates the complexity of decision-making in practice. Ethical passivity may be deleterious to patients' welfare, and concerns emerge about improving pharmacists' ethical training and promoting ethical awareness and responsibility. PMID- 18511617 TI - Moral principles and medical practice: the role of patient autonomy in the extensive use of radiological services. AB - There has been a significant increase in the use of radiological services in the past 30 years. There are many reasons for this, but one has received little attention: the increased role of patient autonomy in healthcare. Patients demand x rays, CT scans, MRI, and positron emission tomography scans. The key question in this article is how a moral principle, such as respect for patient autonomy, can influence the extension of radiological services. A literature review reveals how patient autonomy is acknowledged in radiology, and how it is used both to explain and to justify the increase in radiological examinations. Furthermore, it also shows how the premises favouring patients' exercise of their autonomy are not always present, which makes patient autonomy subject to adverse side effects and even abuse. Patient autonomy can be used to reduce the professionals' responsibility for radiological examinations (by avoiding complaints and lawsuits), to increase the popularity of the profession (by giving the people what they want), to increase the income of the professionals or their institutions, and to promote professional activity. Patient autonomy intended to reduce paternalism, to legitimise otherwise morally unjustifiable actions (such as exposure to radiation), and to protect patients, can easily be used as a moral means for opposite ends. These adverse effects are not peculiar to radiology. However, they emerge particularly clearly in explanations and justifications of the substantial increase in radiological services, as well as in debates on overuse of radiological services. PMID- 18511618 TI - Croatian physicians' and nurses' experience with ethical issues in clinical practice. AB - AIM: To assess ethical issues in everyday clinical practice among physicians and nurses of the University Hospital Rijeka, Rijeka, Croatia. SUBJECTS AND METHODS: We surveyed the entire population of internal medicine, oncology and intensive care specialists and associated nurses employed at the University Hospital Rijeka, Rijeka, Croatia (n = 532). An anonymous questionnaire was used to explore the type and frequency of ethical dilemmas, rank of their difficulty, access to and use of ethics support services, training in ethics and confidence about knowledge in ethics. Physicians (n = 113, 55% of them female) ranged in age from 27 to 61 years, and nurses (n = 251, 95% female), from 20 to 52. RESULTS: The most frequent ethical dilemmas concerned uncertain or impaired decision-making capacity (66% of physicians, 47% of nurses, p = 0.008), limitation of treatment at the end of life (60% of physicians, 31% of nurses, p<0.001) and disagreements among family members (47% of physicians, 31% of nurses, p = 0.025). The most difficult dilemmas concerned euthanasia and physician-assisted suicide (49% of physicians, 52% of nurses) and limitation of treatment at the end of life (14% of physicians, 18% of nurses). Only a minority reported ever using any kind of ethics support services (12% of physicians, 3% of nurses, p = 0.001) or being very confident about knowledge in ethics (5% of physicians, 6% of nurses). CONCLUSIONS: Similar ethical difficulties are present in the clinical practice of both physicians and nurses, with important differences in access and use of ethics support services. A need for systematic ethics educational activities was identified. Inclusion of individual ethics consultants in Croatian healthcare ethics support services is strongly advised. PMID- 18511619 TI - Ethics and the pitchside physician. AB - Sports medicine is a rapidly expanding area of clinical practice. The pitchside physician faces many ethical challenges in managing the injuries and wellbeing of sports men and women. This brief essay describes some of the dilemmas frequently encountered by the pitchside physician and calls for a discussion of the issues raised. PMID- 18511620 TI - Caring for risky patients: duty or virtue? AB - The emergence several years ago of SARS, with its high rate of infection and death among healthcare workers, resurrected a recurring ethical question: do health professionals have a duty to provide care to patients with deadly infectious diseases, even at some substantial risk to themselves and their families? The conventional answer, repeated on the heels of the SARS epidemic, is that they do. In this paper, I argue that the arguments in support of such a duty are wanting in significant respects, and that the language of duty is simply not adequate to an understanding of all the moral dimensions of professional responses to the care of risky patients. Instead, we should speak the language of virtues and ideals if we want to do justice to the complexity of such harrowing circumstances. PMID- 18511621 TI - A pilot study of bullying and harassment among medical professionals in Pakistan, focussing on psychiatry: need for a medical ombudsman. AB - BACKGROUND: The magnitude of bullying and harassment among psychiatrists is reportedly high, yet no peer-review published studies addressing this issue could be found. Therefore, it was decided to conduct a pilot study to assess the degree of the problem, the types of bullying/harassment and to provide some insights into the situation. METHODS AND PRINCIPAL FINDINGS: Following multiple focus group meetings, a yes/no response type questionnaire was developed to assess the degree and type of bullying and harassment experienced by psychiatrists. Over a 3 month period the questionnaire was administered to a random sample of 60 psychiatrists. 57 out of the 60 psychiatrists reported harassment and bullying. Frequencies of the following response variables are presented in descending order: rumours 40% (n = 24); defamation 20% (n = 12); passing remarks 20% (n = 12); false accusations 15% (n = 9); threats 13.3% (n = 8); verbal abuse 13.3% (n = 8); unjustified complaints 13.3% (n = 8); promotion blocked 13.3% (n = 8); humiliation 13% (n = 8); bad reference given 10% (n = 6); credentials questioned 8.3% (n = 5); physical attacks 5% (n = 3); termination 5% (n = 3); derogatory remarks 1.7% (n = 1) and 1.7% (n = 1) were subjected to personal work. As a result of being subjected to harassment, 66.7% of the psychiatrists did not take any action, whereas 33.3% confronted the person(s) they believed responsible. Asked whether the bullying and harassment caused distress, 18.3% of the psychiatrists did not report any effect, 30% reported mild distress, 40% moderate distress and severe distress was reported by 11.7%. CONCLUSIONS: It was concluded that the magnitude of bullying and harassment among psychiatrists may be quite high, as evidenced by this pilot study. There is a need for extensive systematic studies on this subject and to establish strategies to prevent and address this issue at a national and regulatory level. PMID- 18511622 TI - "Eugenics talk" and the language of bioethics. AB - In bioethical discussions of preimplantation genetic diagnosis and prenatal screening, accusations of eugenics are commonplace, as are counter-claims that talk of eugenics is misleading and unhelpful. This paper asks whether "eugenics talk", in this context, is legitimate and useful or something to be avoided. It also looks at the extent to which this linguistic question can be answered without first answering relevant substantive moral questions. Its main conclusion is that the best and most non-partisan argument for avoiding eugenics talk is the Autonomy Argument. According to this, eugenics talk per se is not wrong, but there is something wrong with using its emotive power as a means of circumventing people's critical-rational faculties. The Autonomy Argument does not, however, tell against eugenics talk when such language is used to shock people into critical-rational thought. These conclusions do not depend on unique features of eugenics: similar considerations apply to emotive language throughout bioethics. PMID- 18511623 TI - Systematic reviews of empirical bioethics. AB - BACKGROUND: Publications and discussions of survey research in empirical bioethics have steadily increased over the past two decades. However, findings often differ among studies with similar research questions. As a consequence, ethical reasoning that considers only parts of the existing literature and does not apply systematic reviews tends to be biased. To date, we lack a systematic review (SR) methodology that takes into account the specific conceptual and practical challenges of empirical bioethics. METHODS: The steps of systematically reviewing empirical findings in bioethics are presented and critically discussed. In particular, (a) the limitations of traditional SR methodologies in the field of empirical bioethics are critically discussed, and (b) conceptual and practical recommendations for SRs in empirical bioethics that are (c) based on the authors' review experiences in healthcare ethics are presented. RESULTS: A 7-step approach for SRs of empirical bioethics is proposed: (1) careful definition of review question; (2) selection of relevant databases; (3) application of ancillary search strategies; (4) development of search algorithms; (5) relevance assessment of the retrieved references; (6) quality assessment of included studies; and (7) data analysis and presentation. Conceptual and practical challenges arise because of various peculiarities in reviewing empirical bioethics literature and can lead to biased results if they are not taken into account. CONCLUSIONS: If suitably adapted to the peculiarities of the field, SRs of empirical bioethics provide transparent information for ethical reasoning and decision-making that is less biased than single studies. PMID- 18511624 TI - National HIV treatment guidelines in Tanzania and Ethiopia: are they legitimate rationing tools? AB - OBJECTIVE: To provide an ethical analysis of whether the Ethiopian and Tanzanian national HIV/AIDS treatment guidelines can be considered legitimate and fair rationing tools. METHOD: Qualitative study and ethical analysis involving guideline documents and interviews with nine key members involved in the development of the guidelines. The analysis followed an editing organising style. The theoretical framework was a guideline-specific framework based on theories of just resource allocation in healthcare and conditions that ensure fair processes in guideline development. According to this framework, legitimate rationing requires reasons for patient selection to be explicit, public and relevant, and decisions must be open to question and revision. RESULTS: The only explicit rationing criteria that both guidelines recommended were clinical antiretroviral treatment indications. Explicit non-clinical rationing criteria were expressed in a separate Ethiopian implementation guideline. Neither of the guideline development processes fully satisfies minimal requirements of procedural fairness. There is a lack of transparency. The reasons for decisions are rarely given and are not publicly available. This reduces the opportunity for public questioning, debate and revisions. The guidelines were based on expert opinion and consensus. Recommendations from the WHO were copied without much discussion, disagreement or adjustment. CONCLUSIONS: The two national HIV treatment guidelines discussed are de facto mechanisms for rationing but were developed using methods that do not fully satisfy the requirements of fair processes. PMID- 18511625 TI - "Living apart together": moral frictions between two coexisting organ transplantation schemes. AB - Cadaveric transplantation and living transplantation exist side by side. Both practices help to alleviate organ need. They provide us with two separate moral schemes. Is it rational to keep them apart? The cadaveric system is organised along strict, impartial lines, while the living system is inherently partial and local. The ethical justification for this partial scheme seems to be that it merely supplements the cadaveric scheme: partial transplants do not come at the expense of cadaveric impartiality, but in fact significantly reduce the waiting time for patients on the list for a cadaveric transplant. This seemingly peaceful coexistence is challenged by new initiatives, among them living donation list exchange, and also the LifeSharers initiative, leading to practices that undermine cadaveric impartiality. Should we bemoan this fact, or should we move on towards a new balance in the relationship between cadaveric and living transplantation practices, towards a new moral weighing of impartial and partial values? I argue, against the background of a rapid growth of living donations, that we have good, ethical reasons--not only utilitarian ones--for giving the value of partiality a more prominent place in our policies. PMID- 18511626 TI - The experiences of ethics committee members: contradictions between individuals and committees. AB - The current system of ethical review for medical research in the United Kingdom is changing from the current system involving large committees of 7-18 members reviewing every individual application to a system involving pre-review by small sub-committees of National Research Ethics Officers (NREAs), who have a remit to approve studies if they believe there are no material ethical issues imposed by the research. The reliability of this new system depends on the reliability of the NREAs and in particular the ability of small groups to effectively identify and appropriately assess the seriousness of all the material ethical issues that may be posed by an application. There is anecdotal evidence to suggest that many individual research ethics committee members have had experience of believing that a study presents no material ethical concerns, then on reaching the committee and discussing the application they realise that the committee feels it does present significant ethical concerns. If this is the case then this casts doubt on the reliability of NREAs or small groups to effectively identify ethically problematic research and appropriately respond to this to protect research participants. In this paper we describe a small questionnaire based piece of research carried out to assess how common this and other relevant experiences are. PMID- 18511627 TI - Genes, race and research ethics: who's minding the store? AB - BACKGROUND: The search for genetic variants between racial/ethnic groups to explain differential disease susceptibility and drug response has provoked sharp criticisms, challenging the appropriateness of using race/ethnicity as a variable in genetics research, because such categories are social constructs and not biological classifications. OBJECTIVES: To gain insight into how a group of genetic scientists conceptualise and use racial/ethnic variables in their work and their strategies for managing the ethical issues and consequences of this practice. METHODS: In-depth semi-structured interviews were conducted with a purposive sample of 30 genetic researchers who use racial/ethnic variables in their research. Standard qualitative methods of content analysis were used. RESULTS: Most of the genetic researchers viewed racial/ethnic variables as arbitrary and very poorly defined, and in turn as scientifically inadequate. However, most defended their use, describing them as useful proxy variables on a road to "imminent medical progress". None had developed overt strategies for addressing these inadequacies, with many instead asserting that science will inevitably correct itself and saying that meanwhile researchers should "be careful" in the language chosen for reporting findings. CONCLUSIONS: While the legitimacy and consequences of using racial/ethnic variables in genetics research has been widely criticised, ethical oversight is left to genetic researchers themselves. Given the general vagueness and imprecision we found amongst these researchers regarding their use of these variables, they do not seem well equipped for such an undertaking. It would seem imperative that research ethicist move forward to develop specific policies and practices to assure the scientific integrity of genetic research on biological differences between population groups. PMID- 18511628 TI - What to tell and how to tell: a qualitative study of information sharing in research for adults with intellectual disability. AB - OBJECTIVES: To explore opinions and attitudes regarding the current information giving practices in research involving adults with intellectual disabilities. DESIGN: Qualitative focus group study with a purposive sample. SETTING: An intellectual disabilities service within the NHS PARTICIPANTS: A sample of 26 individuals including adults with mild intellectual disability, carers, clinicians, care managers and the charitable sector. RESULTS: Three main themes were identified: process, format, and content. There was agreement that there is a need for improvement in the process and quality of information giving. With regard to the content of information, there was little discrepancy between the study findings and existing guidance. CONCLUSIONS: Information should be presented in different formats and prepared with input from appropriate professions. Additionally the roles of peers, carers and other professionals in the process of information giving should be considered. PMID- 18511629 TI - Scientific responsibility for the dissemination and interpretation of genetic research: lessons from the "warrior gene" controversy. AB - This paper discusses the announcement by a team of researchers that they identified a genetic influence for a range of "antisocial" behaviours in the New Zealand Maori population (dubbed the "warrior gene"). The behaviours included criminality, violence, gambling and alcoholism. The reported link between genetics and behaviour met with much controversy. The scientists were described as hiding behind a veneer of supposedly "objective" western science, using it to perpetuate "racist and oppressive discourses". In this paper we examine what went wrong in the dissemination of the research. We chose as our framework the debate around the "internal/external" responsibilities of scientists. Using this discourse we argue that when the researchers ventured to explain their research in terms of social phenomena, they assumed a duty to ensure that their findings were placed "in context". By "in context", we argue that evidence of any genetic influence on behavioural characteristics should not be reported in isolation, but instead presented alongside other environmental, cultural and socio-economic influences that may also contribute to the studied behaviour. Rather than imposing a new obligation on scientists, we find this duty to contextualise results is in keeping with the spirit of codes of ethics already in place. Lessons from the "warrior gene" controversy may assist researchers elsewhere to identify potential areas of conflict before they jeopardise research relationships, or disseminate findings in a manner that fuels misleading and/or potentially discriminatory attitudes in society. PMID- 18511632 TI - Should we stop using intravenous gentamicin in patients with cystic fibrosis? PMID- 18511633 TI - Driving and obstructive sleep apnoea. PMID- 18511634 TI - Prenatal nutrition and asthma: hope or hype? PMID- 18511635 TI - Aspirin and asthma: barking up the right tree? PMID- 18511636 TI - Progressive dyspnoea, pleural effusions and lytic bone lesions. PMID- 18511640 TI - Genetic susceptibility to the respiratory effects of air pollution. AB - There is large variation between individuals in their response to air pollutants. This review summarises the existing evidence that genetic factors influence the mechanisms of lung injury caused by air pollutants. Genetic association studies have compared the adverse effects of air pollutants between subjects with specific genotypes in biologically relevant genes. In human studies of ozone exposure, polymorphisms in oxidative stress genes (NQO1, GSTM1, GSTP1) modify respiratory symptoms, lung function, biomarkers and risk of asthma. Inflammatory gene polymorphisms (TNF) influence the lung function response to ozone, and the effect of different levels of ozone on the development of asthma. Polymorphisms in oxidative stress genes (GSTM1, GSTP1) alter the response to combined exposure to ragweed pollen and diesel exhaust particles. Importantly, polymorphisms in an oxidative stress gene (GSTM1) have predicted patients with asthma who benefit from antioxidant supplementation in Mexico City, which has chronically high ozone exposure. Genetic linkage studies of families have not been feasible for studying the effects of air pollution in humans, but some progress has been made with pedigrees of specially bred mice, in identifying chromosomal regions linked to effects of ozone or particles. A high priority now, in addition to avoiding exposure in the most susceptible people, is to clearly identify the most effective and safe chemopreventive agents for individuals who are genetically susceptible to the adverse effects of air pollution (eg, antioxidants to be taken during high ozone levels). PMID- 18511641 TI - Intrabullous blood injection for lung volume reduction. AB - Bronchoscopic treatment for emphysematous lung diseases has attracted clinical attention, and several different approaches are being investigated. We present a case of emphysematous bullae that was effectively treated with a newly developed bronchoscopic intervention, autologous blood injection. A 59-year-old man was referred to our institution with exertional dyspnoea. Chest CT showed emphysema and bullae with a diameter of 12 cm in the right upper lobe. Bronchoscopic treatment was introduced as an alternative to surgery. Autologous blood and fibrinogen solution were infused into bullae via the transbronchial catheter, under fluoroscopic guidance. Post-treatment CT showed marked contraction of bullae to a diameter of 3 cm, corresponding to a volume reduction of 800 ml on body plethysmography. A significant reduction in dyspnoea was also noted. This therapeutic approach is less invasive and may represent a good option for reducing lung volume. PMID- 18511642 TI - Gene expression of IL17 and IL23 in the lungs of patients with active tuberculosis. PMID- 18511643 TI - Early detection of cancer: lessons from lung cancer CT screening. PMID- 18511644 TI - An integrated home oxygen service saves pound sterling 130 000 per year on home oxygen tariffs. PMID- 18511645 TI - Use of long-acting beta2 agonists in arginine-16 homozygous patients with asthma. PMID- 18511646 TI - Dysfunctional breathing in patients with asthma. PMID- 18511647 TI - Accuracy in suspicious lung infiltrations. PMID- 18511648 TI - Delayed-onset bronchiolitis obliterans with organising pneumonia associated with massive acetic acid steam inhalation. PMID- 18511649 TI - Behavioral pharmacology of the mu/delta opioid glycopeptide MMP2200 in rhesus monkeys. AB - H(2)N-Tyr-D-Thr-Gly-Phe-Leu-Ser-(O-beta-D-lactose)-CONH(2) (MMP2200) is a novel glycopeptide opioid agonist with similar affinities for mu and delta receptors. Glycosylation promoted brain penetration and production of centrally mediated behavioral effects in mice; however, it is unknown whether the magnitude of enhanced brain penetration is sufficient to permit central mediation of drug effects and production of synergistic mu/delta antinociceptive interactions after systemic administration in primates. To address this issue, the present study compared the effects of MMP2200 and the mu-agonist morphine in four behavioral procedures in rhesus monkeys. In an assay of thermal nociception, morphine (1.0 5.6 mg/kg) produced dose-dependent antinociception, whereas MMP2200 (10-56 mg/kg) was ineffective. In an assay of capsaicin-induced thermal allodynia, both morphine (0.01-1.0 mg/kg) and MMP2200 (0.032-3.2 mg/kg) produced dose-dependent antiallodynic effects. MMP2200-induced antiallodynia was blocked by the moderately mu-selective antagonist naltrexone (0.01 mg/kg), the delta-selective antagonist naltrindole (1.0 mg/kg), and the peripherally selective opioid antagonist quaternary naltrexone (0.32 mg/kg). In an assay of schedule-controlled behavior, both morphine (0.01-1.0 mg/kg) and MMP2200 (10-56 mg/kg) decreased response rates. Morphine effects were antagonized by naltrexone (0.001-0.01 mg/kg); however, the effects of MMP2200 were not antagonized by either naltrexone (0.01 mg/kg) or naltrindole (1.0 mg/kg). In an assay of drug self-administration, morphine (0.0032-0.32 mg/kg/injection) produced reinforcing effects, whereas MMP2200 (0.032-0.32 mg/kg/injection) did not. These results suggest that systemically administered MMP2200 acted as a peripheral, mu/delta-opioid agonist with limited distribution to the central nervous system in rhesus monkeys. These results also suggest the existence of species differences in the pharmacokinetics and brain penetration of glycopeptides. PMID- 18511650 TI - Pharmacokinetics of high-dose abetimus sodium in normal subjects with specific assessment of effect on coagulation. AB - Abetimus sodium is an oligonucleotide-based investigational drug designed to treat patients with lupus nephritis by specifically reducing anti-double-stranded DNA antibody levels. The safety and pharmacokinetics of abetimus were evaluated in 24 healthy volunteers at intravenous doses of 600 mg, 1200 mg, and 2400 mg. The mean half-life ranged from 0.8 to 1.5 hours. Maximum exposure assessed by maximum observed plasma concentration was dose proportional. Total exposure assessed by area under the plasma concentration-time curve was dose proportional between 1200-mg and 2400-mg doses and greater than proportionate between the 600 mg and 1200-mg doses. Abetimus was well tolerated in all dose groups, with adverse events observed in 33.3% (2/6) of placebo subjects and 16.7% (3/18) subjects receiving abetimus. No clinically significant effects were observed on laboratory values, vital signs, or electrocardiogram, with the exception of a transient, dose-dependent, prolongation in activated partial thromboplastin time. In vitro coagulation studies suggested that the effect on activated partial thromboplastin time was attributable to a nonspecific interaction rather than specific factor depletion. Exposure to abetimus at intravenous doses of 600 mg, 1200 mg, and 2400 mg was well tolerated. Total exposure assessed by area under the plasma concentration-time curve was greater than dose proportional across the dose range of 600 mg to 2400 mg. PMID- 18511651 TI - Probenecid, but not cystic fibrosis, alters the total and renal clearance of fexofenadine. AB - This study aims to evaluate renal P-glycoprotein (P-gp) activity in patients with cystic fibrosis. P-gp efflux activity in peripheral T cells was measured by flow cytometry in 10 cystic fibrosis and 15 healthy volunteers. Eight cystic fibrosis patients and 8 healthy volunteers were recruited into a crossover pharmacokinetic study in which participants received 180 mg fexofenadine with or without 1 g probenecid twice a day. Genotyping was performed for ABCB1 C1236T, G2677T, and C3435T. P-gp efflux activity in peripheral T cells was not significantly different between cystic fibrosis patients and healthy volunteers. No difference in fexofenadine pharmacokinetic parameters was observed between cystic fibrosis patients and healthy volunteers when fexofenadine was administered with or without probenecid. Coadministration of probenecid significantly increased fexofenadine AUC and decreased the cumulative urinary excretion, total body clearance, and renal clearance. ABCB1 3435 C/T carriers showed increased basal P gp activity in CD4+ and CD8+ T cells, increased R123-induced efflux activity in CD4+ T cell, and decreased fexofenadine AUC. Fexofenadine disposition and P-gp efflux activity in peripheral T cells was similar between cystic fibrosis patients and healthy volunteers. Probenecid administration significantly reduced the total body and renal clearance of fexofenadine. ABCB1 3435 C/T was associated with an elevated efflux activity compared with C/C subjects. PMID- 18511653 TI - Protein-protein interactions and dopamine D2 receptor signaling: a calcium connection. AB - The third cytoplasmic loop is a crucial site of physical contact between some G protein-coupled receptors (GPCRs) and their respective G proteins. However, interactions not only occur among these proteins but also involve a number of additional protein binding partners. Modulation of these protein-protein interactions may represent an important new avenue of pharmacotherapy through which signaling of GPCRs can be modulated. In the current issue of Molecular Pharmacology, Liu et al. (p. 371) report that dopamine D(2) receptors interact with the Ca(2+) binding protein S100B. Using the third intracellular loop of the dopamine D(2) receptor as bait in a bacterial two-hybrid system, S100B was determined to be a potential binding partner. They used pull-down assays both in vitro and in vivo to confirm the interaction and define its specificity. Neither the D(3) nor the D(4) receptor expresses the motif conferring the interaction, and peptides based on the third intracellular loop of the D(3) receptor did not pull down S100B. Some groups might stop there, but Liu and colleagues moved on to demonstrate colocalization of the D(2) receptor and S100B by immunostaining. Functional assays were then used to show that coexpression of S100B with the D(2) receptor increases the ability of D(2) receptors to activate ERK and to inhibit adenylyl cyclase. These data suggest that S100B coexpression may serve as an important mediator of D(2) receptor signaling efficacy in the brain. These interactions contribute to cellular, regional, and developmental differences in D(2) receptor activation. PMID- 18511652 TI - Sulfonylurea receptor-dependent and -independent pathways mediate vasodilation induced by ATP-sensitive K+ channel openers. AB - ATP-sensitive K+ (KATP) channel openers are vasodilators that activate both plasma membrane and mitochondrial KATP channels. Here, we investigated the molecular mechanisms by which diazoxide and pinacidil induce vasodilation by studying diameter regulation of wild-type [SUR2(+/+)] and sulfonylurea receptor (SUR) 2-deficient [SUR2(-/-)] mouse myogenic mesenteric arteries. Ryanodine (10 microM), a ryanodine-sensitive Ca2+ release (RyR) channel blocker; iberiotoxin (100 nM), a large-conductance Ca2+-activated K+ (KCa) channel blocker; 4 aminopyridine (4-AP; 1 mM), a voltage-gated K+ (KV) channel blocker; manganese(III) tetrakis(1-methyl-4-pyridyl)porphyrin (MnTMPyP; 100 microM), an antioxidant; and a combination of ryanodine and 4-AP reduced diazoxide (100 microM)-induced dilation in pressurized (60 mm Hg) SUR2(+/+) arteries by 45 to 77%. In contrast, these inhibitors did not alter pinacidil (5 microM)-induced dilation in SUR2(+/+) arteries. Reverse transcription-polymerase chain reaction indicated that SUR2B was the only SUR isoform expressed in SUR2(+/+) mesenteric artery smooth muscle cells, whereas SURs were absent in SUR2(-/-) cells. In SUR2( /-) arteries, pinacidil-induced vasodilation was 10% of that in SUR2(+/+) arteries, whereas diazoxide-induced vasodilation was similar in SUR2(+/+) and SUR2(-/-) arteries. Atpenin (1 microM), a selective electron transport chain (ETC) complex II inhibitor, dilated arteries similarly to diazoxide, and this effect was attenuated by MnTMPyP and ryanodine + 4-AP. Atpenin also attenuated diazoxide-, but not pinacidil-induced vasodilation. In summary, data indicate that pinacidil-induced vasodilation requires SUR2B, whereas diazoxide-induced vasodilation does not require SURs. Rather, diazoxide-induced vasodilation involves ETCII inhibition; a smooth muscle cell-reactive oxygen species elevation; and RyR, KCa, and KV channel activation. These data indicate that KATP channel openers regulate arterial diameter via SUR-dependent and -independent pathways. PMID- 18511654 TI - Paleo-Eskimo mtDNA genome reveals matrilineal discontinuity in Greenland. AB - The Paleo-Eskimo Saqqaq and Independence I cultures, documented from archaeological remains in Northern Canada and Greenland, represent the earliest human expansion into the New World's northern extremes. However, their origin and genetic relationship to later cultures are unknown. We sequenced a mitochondrial genome from a Paleo-Eskimo human by using 3400-to 4500-year-old frozen hair excavated from an early Greenlandic Saqqaq settlement. The sample is distinct from modern Native Americans and Neo-Eskimos, falling within haplogroup D2a1, a group previously observed among modern Aleuts and Siberian Sireniki Yuit. This result suggests that the earliest migrants into the New World's northern extremes derived from populations in the Bering Sea area and were not directly related to Native Americans or the later Neo-Eskimos that replaced them. PMID- 18511655 TI - Structural basis of trans-inhibition in a molybdate/tungstate ABC transporter. AB - Transport across cellular membranes is an essential process that is catalyzed by diverse membrane transport proteins. The turnover rates of certain transporters are inhibited by their substrates in a process termed trans-inhibition, whose structural basis is poorly understood. We present the crystal structure of a molybdate/tungstate ABC transporter (ModBC) from Methanosarcina acetivorans in a trans-inhibited state. The regulatory domains of the nucleotide-binding subunits are in close contact and provide two oxyanion binding pockets at the shared interface. By specifically binding to these pockets, molybdate or tungstate prevent adenosine triphosphatase activity and lock the transporter in an inward facing conformation, with the catalytic motifs of the nucleotide-binding domains separated. This allosteric effect prevents the transporter from switching between the inward-facing and the outward-facing states, thus interfering with the alternating access and release mechanism. PMID- 18511656 TI - Elevation changes in Antarctica mainly determined by accumulation variability. AB - Antarctic Ice Sheet elevation changes, which are used to estimate changes in the mass of the interior regions, are caused by variations in the depth of the firn layer. We quantified the effects of temperature and accumulation variability on firn layer thickness by simulating the 1980-2004 Antarctic firn depth variability. For most of Antarctica, the magnitudes of firn depth changes were comparable to those of observed ice sheet elevation changes. The current satellite observational period ( approximately 15 years) is too short to neglect these fluctuations in firn depth when computing recent ice sheet mass changes. The amount of surface lowering in the Amundsen Sea Embayment revealed by satellite radar altimetry (1995-2003) was increased by including firn depth fluctuations, while a large area of the East Antarctic Ice Sheet slowly grew as a result of increased accumulation. PMID- 18511657 TI - Climate change. A Matter of firn. PMID- 18511660 TI - Physics. Energy department pulls plug on overbudget fusion experiment. PMID- 18511658 TI - Probing cold dense nuclear matter. AB - The protons and neutrons in a nucleus can form strongly correlated nucleon pairs. Scattering experiments, in which a proton is knocked out of the nucleus with high momentum transfer and high missing momentum, show that in carbon-12 the neutron proton pairs are nearly 20 times as prevalent as proton-proton pairs and, by inference, neutron-neutron pairs. This difference between the types of pairs is due to the nature of the strong force and has implications for understanding cold dense nuclear systems such as neutron stars. PMID- 18511659 TI - Education to protect humanity. PMID- 18511661 TI - Parkinson's disease. Streamlined clinical trials, from a home computer. PMID- 18511662 TI - Wenchuan earthquake. Damaged university mourns its dead--and plans fast recovery. PMID- 18511663 TI - Biosecurity. Fate of Plum Island animal lab still unclear. PMID- 18511664 TI - Genetics. Ancient DNA from frozen hair may untangle Eskimo roots. PMID- 18511665 TI - Epidemiology. Children's study needs pilot testing, panel finds. PMID- 18511666 TI - Particle physics. Does Fermilab have a future? PMID- 18511667 TI - Particle physics. Whither the International Linear Collider? PMID- 18511668 TI - Antarctica. Freeze-dried findings support a tale of two ancient climates. PMID- 18511669 TI - Medicine. Nothing rotten about hydrogen sulfide's medical promise. PMID- 18511670 TI - Joshua Lederberg's interest in ignorance. PMID- 18511671 TI - Women in science: a top-down approach. PMID- 18511672 TI - Biological basis of the third-cousin crush. PMID- 18511673 TI - Comment on "Mixed-layer deepening during Heinrich events: a multi-planktonic foraminiferal delta18O approach". AB - Rashid and Boyle (Reports, 19 October 2007, p. 439) analyzed oxygen isotopes in planktonic foraminera from marine sediments and concluded that Heinrich events (massive iceberg discharges into the North Atlantic Ocean) caused upper water masses to deepen. We question the robustness of this interpretation and argue that a strongly stratified mixed layer characterized by dense sea-ice cover and production of oxygen-18-depleted brines likely prevailed during such events. PMID- 18511674 TI - Diversity. Culture, gender, and math. PMID- 18511675 TI - Atmospheric science. Whither geoengineering? PMID- 18511676 TI - Astronomy. A blast from the past. PMID- 18511677 TI - Immunology. Immunity benefits from a little suppression. PMID- 18511678 TI - Materials science. Graphene-based materials. PMID- 18511679 TI - Planetary science. Is Mars geodynamically dead? PMID- 18511680 TI - Medicine. Activating a repressor. PMID- 18511681 TI - How we see ourselves and how we see others. AB - People see themselves differently from how they see others. They are immersed in their own sensations, emotions, and cognitions at the same time that their experience of others is dominated by what can be observed externally. This basic asymmetry has broad consequences. It leads people to judge themselves and their own behavior differently from how they judge others and those others' behavior. Often, those differences produce disagreement and conflict. Understanding the psychological basis of those differences may help mitigate some of their negative effects. PMID- 18511682 TI - Ecosystem feedbacks and nitrogen fixation in boreal forests. AB - Biological feedback mechanisms regulate fundamental ecosystem processes and potentially regulate ecosystem productivity. To date, no studies have documented the down-regulation of terrestrial nitrogen (N) fixation via an ecosystem-level feedback mechanism. Herein, we demonstrate such a feedback in boreal forests. Rapid cycling of N in early secondary succession forests yielded greater throughfall N deposition, which in turn decreased N fixation by cyanobacterial associates in feather moss carpets that reside on the forest floor. The forest canopy exerts a tight control on biotic N input at a period of high productivity. PMID- 18511683 TI - Predicting human brain activity associated with the meanings of nouns. AB - The question of how the human brain represents conceptual knowledge has been debated in many scientific fields. Brain imaging studies have shown that different spatial patterns of neural activation are associated with thinking about different semantic categories of pictures and words (for example, tools, buildings, and animals). We present a computational model that predicts the functional magnetic resonance imaging (fMRI) neural activation associated with words for which fMRI data are not yet available. This model is trained with a combination of data from a trillion-word text corpus and observed fMRI data associated with viewing several dozen concrete nouns. Once trained, the model predicts fMRI activation for thousands of other concrete nouns in the text corpus, with highly significant accuracies over the 60 nouns for which we currently have fMRI data. PMID- 18511684 TI - The Cassiopeia A supernova was of type IIb. AB - Cassiopeia A is the youngest supernova remnant known in the Milky Way and a unique laboratory for supernova physics. We present an optical spectrum of the Cassiopeia A supernova near maximum brightness, obtained from observations of a scattered light echo more than three centuries after the direct light of the explosion swept past Earth. The spectrum shows that Cassiopeia A was a type IIb supernova and originated from the collapse of the helium core of a red supergiant that had lost most of its hydrogen envelope before exploding. Our finding concludes a long-standing debate on the Cassiopeia A progenitor and provides new insight into supernova physics by linking the properties of the explosion to the wealth of knowledge about its remnant. PMID- 18511685 TI - Interfacial polygonal nanopatterning of stable microbubbles. AB - Micrometer-sized bubbles are unstable and therefore difficult to make and store for substantial lengths of time. Short-term stabilization is achieved by the addition of amphiphilic molecules, which reduce the driving force for dissolution. When these molecules crystallize on the air/liquid interface, the lifetime of individual bubbles may extend over a few months. We demonstrated low gas-fraction dispersions with mean bubble radii of less than 1 micrometer and stability lasting more than a year. An insoluble, self-assembled surfactant layer covers the surface of the microbubbles, which can result in nanometer-scale hexagonal patterning that we explain with thermodynamic and molecular models. The elastic response of the interface arrests the shrinkage of the bubbles. Our study identifies a route to fabricate highly stable dispersions of microbubbles. PMID- 18511686 TI - Water activity and the challenge for life on early Mars. AB - In situ and orbital exploration of the martian surface has shown that acidic, saline liquid water was intermittently available on ancient Mars. The habitability of these waters depends critically on water activity (aH2O), a thermodynamic measure of salinity, which, for terrestrial organisms, has sharply defined limits. Using constraints on fluid chemistry and saline mineralogy based on martian data, we calculated the maximum aH2O for Meridiani Planum and other environments where salts precipitated from martian brines. Our calculations indicate that the salinity of well-documented surface waters often exceeded levels tolerated by known terrestrial organisms. PMID- 18511687 TI - A cytosolic iron chaperone that delivers iron to ferritin. AB - Ferritins are the main iron storage proteins found in animals, plants, and bacteria. The capacity to store iron in ferritin is essential for life in mammals, but the mechanism by which cytosolic iron is delivered to ferritin is unknown. Human ferritins expressed in yeast contain little iron. Human poly (rC) binding protein 1 (PCBP1) increased the amount of iron loaded into ferritin when expressed in yeast. PCBP1 bound to ferritin in vivo and bound iron and facilitated iron loading into ferritin in vitro. Depletion of PCBP1 in human cells inhibited ferritin iron loading and increased cytosolic iron pools. Thus, PCBP1 can function as a cytosolic iron chaperone in the delivery of iron to ferritin. PMID- 18511688 TI - Massive horizontal gene transfer in bdelloid rotifers. AB - Horizontal gene transfer in metazoans has been documented in only a few species and is usually associated with endosymbiosis or parasitism. By contrast, in bdelloid rotifers we found many genes that appear to have originated in bacteria, fungi, and plants, concentrated in telomeric regions along with diverse mobile genetic elements. Bdelloid proximal gene-rich regions, however, appeared to lack foreign genes, thereby resembling those of model metazoan organisms. Some of the foreign genes were defective, whereas others were intact and transcribed; some of the latter contained functional spliceosomal introns. One such gene, apparently of bacterial origin, was overexpressed in Escherichia coli and yielded an active enzyme. The capture and functional assimilation of exogenous genes may represent an important force in bdelloid evolution. PMID- 18511689 TI - Ancestral monogamy shows kin selection is key to the evolution of eusociality. AB - Close relatedness has long been considered crucial to the evolution of eusociality. However, it has recently been suggested that close relatedness may be a consequence, rather than a cause, of eusociality. We tested this idea with a comparative analysis of female mating frequencies in 267 species of eusocial bees, wasps, and ants. We found that mating with a single male, which maximizes relatedness, is ancestral for all eight independent eusocial lineages that we investigated. Mating with multiple males is always derived. Furthermore, we found that high polyandry (>2 effective mates) occurs only in lineages whose workers have lost reproductive totipotency. These results provide the first evidence that monogamy was critical in the evolution of eusociality, strongly supporting the prediction of inclusive fitness theory. PMID- 18511690 TI - Log or linear? Distinct intuitions of the number scale in Western and Amazonian indigene cultures. AB - The mapping of numbers onto space is fundamental to measurement and to mathematics. Is this mapping a cultural invention or a universal intuition shared by all humans regardless of culture and education? We probed number-space mappings in the Mundurucu, an Amazonian indigene group with a reduced numerical lexicon and little or no formal education. At all ages, the Mundurucu mapped symbolic and nonsymbolic numbers onto a logarithmic scale, whereas Western adults used linear mapping with small or symbolic numbers and logarithmic mapping when numbers were presented nonsymbolically under conditions that discouraged counting. This indicates that the mapping of numbers onto space is a universal intuition and that this initial intuition of number is logarithmic. The concept of a linear number line appears to be a cultural invention that fails to develop in the absence of formal education. PMID- 18511691 TI - MeCP2, a key contributor to neurological disease, activates and represses transcription. AB - Mutations in the gene encoding the transcriptional repressor methyl-CpG binding protein 2 (MeCP2) cause the neurodevelopmental disorder Rett syndrome. Loss of function as well as increased dosage of the MECP2 gene cause a host of neuropsychiatric disorders. To explore the molecular mechanism(s) underlying these disorders, we examined gene expression patterns in the hypothalamus of mice that either lack or overexpress MeCP2. In both models, MeCP2 dysfunction induced changes in the expression levels of thousands of genes, but unexpectedly the majority of genes (approximately 85%) appeared to be activated by MeCP2. We selected six genes and confirmed that MeCP2 binds to their promoters. Furthermore, we showed that MeCP2 associates with the transcriptional activator CREB1 at the promoter of an activated target but not a repressed target. These studies suggest that MeCP2 regulates the expression of a wide range of genes in the hypothalamus and that it can function as both an activator and a repressor of transcription. PMID- 18511693 TI - Effects of study task on the neural correlates of source encoding. AB - The present study investigated whether the neural correlates of source memory vary according to study task. Subjects studied visually presented words in one of two background contexts. In each test, subjects made old/new recognition and source memory judgments. In one study test cycle, study words were subjected to animacy judgments, whereas in another cycle the study task required syllable judgments. Functional magnetic resonance imaging (fMRI) was employed to contrast the neural activity elicited by study words that attracted accurate source judgments on the later memory test, as opposed to words for which source judgments were incorrect or for which source information was unavailable. In both tasks, relative to words for which source memory failed, study words that were later assigned to the correct source elicited enhanced activity in ventral extrastriate cortex. In addition to these common effects of subsequent source memory, additional effects were observed that were selective for each study task. The present findings add weight to the proposal that neural activity supporting successful episodic memory encoding is a reflection of both the online processing engaged by an episode as it is experienced, and the demands imposed by the later retrieval task. PMID- 18511694 TI - Activation of LVGCCs and CB1 receptors required for destabilization of reactivated contextual fear memories. AB - Previous studies have shown that inhibiting protein synthesis shortly after reactivation impairs the subsequent expression of a previously consolidated fear memory. This has suggested that reactivation returns a memory to a labile state and that protein synthesis is required for the subsequent restabilization of memory. While the molecular mechanisms underlying the restabilization of reactivated memories are being uncovered, those underlying the initial destabilization are not known at all. Using a contextual fear conditioning paradigm in mice, here we show that LVGCCs or CB1 receptors in hippocampus are required for the initial destabilization of reactivated memory. Either pharmacological blockade of hippocampal protein synthesis or genetic disruption of CREB-dependent transcription disrupts memory restabilization following reactivation. However, these effects were completely blocked when mice were treated with inhibitors of either LVGCCs or CB1 receptors, indicating that LVGCCs or CB1 receptors are required for the initial destabilization of reactivated memory. In control experiments, we show that blockade of LVGCCs or CB1 receptors does not interfere with the ability of ANI to block protein synthesis, or with the ability of ANI to impair initial consolidation. These experiments begin to reveal mechanisms underlying the destabilization of previously consolidated memories following reactivation and indicate the importance of activation of LVGCCs and CB1 in this process. PMID- 18511695 TI - Direct treatment of mouse or human blood with soluble 5'-nucleotidase inhibits platelet aggregation. AB - OBJECTIVE: Adenosine signaling is known to inhibit platelet aggregation. Extracellular adenosine mainly stems from enzymatic phosphohydrolysis of precursor nucleotides via ecto-5'-nucleotidase. Previous studies suggest that soluble 5'-nucleotidase (5'-NT) derived from Crotalus atrox venom may be clinically beneficial in vascular leakage, myocardial, renal, and intestinal ischemia, or acute lung injury. However, the effects of 5'-NT treatment on platelet aggregation remain unknown. We examined the direct effects of 5'-NT treatment on platelet aggregation in vivo and ex vivo using a whole blood aggregation method. METHODS AND RESULTS: Platelet aggregation in whole human blood was completely inhibited by 5'-NT. When 5'-[alphabeta-methylene] diphosphate (APCP), a specific 5'-ecto-nucleotidase inhibitor, was added together with 5'-NT, APCP fully restored collagen- or ADP-induced aggregation. Adenosine levels in whole blood were significantly increased after 5'-NT treatment compared to controls and inhibition of platelet aggregation by 5'-NT was completely reversed by pretreatment with the nonspecific adenosine receptor antagonist 8-(p sulfophenyl)theophylline hydrate (8-SPT), suggesting that 5'-NT inhibits aggregation via increased adenosine signaling. Administration of 5'-NT to mice in vivo abolished ADP- and collagen-induced platelet aggregation and increased adenosine concentrations and tail bleeding time. CONCLUSIONS: 5'-NT treatment inhibits platelet aggregation via generation of increased levels of extracellular adenosine and subsequent adenosine receptor signaling. PMID- 18511696 TI - A novel variant in the platelet endothelial aggregation receptor-1 gene is associated with increased platelet aggregability. AB - OBJECTIVE: Platelet endothelial aggregation receptor-1 (PEAR1) is a recently identified platelet transmembrane protein that becomes activated by platelet contact. We looked for novel genetic variants in PEAR1 and studied their association with agonist-induced native platelet aggregation and with the inhibitory effect of aspirin on platelets. METHODS AND RESULTS: We genotyped PEAR1 for 10 single nucleotide polymorphisms (SNPs), selected for optimal gene coverage at a density of 4 kb, in 1486 apparently healthy individuals from two generations of families with premature CAD. Subjects had a mean age of 45 years; 62% were white and 38% black. Platelet aggregation to collagen, epinephrine, and ADP was measured in platelet rich plasma, at baseline and after 2 weeks of aspirin (ASA, 81 mg/d), and genotype-phenotype associations were examined separately by ethnicity using multivariable generalized linear models adjusted for covariates. The C allele of SNP rs2768759 [A/C], located in the promoter region of the gene, was common in whites and uncommon in blacks (allele frequency 70.2% versus 17.7%). The C allele was generally associated in both ethnic groups with increased aggregation of native platelets to each agonist. After ASA, the associations were stronger and more consistent and remained significant when post ASA aggregation was adjusted for baseline aggregation, consistent with a relationship between the C allele and reduced platelet responsiveness to ASA. The PEAR1 SNP explained up to 6.9% of the locus specific genetic variance in blacks and up to 2.5% of the genetic variance in whites after ASA. CONCLUSIONS: PEAR1 appears to play an important role in agonist-induced platelet aggregation and in the response to ASA in both whites and blacks. PMID- 18511697 TI - Genetic variants of Y chromosome are associated with a protective lipid profile in black men. AB - OBJECTIVE: Gender and ethnicity modulate the phenotypic expression of cardiovascular risk factors. In particular, men are at higher risk of developing cardiovascular diseases compared to women, whereas black populations of African origin display reduced mortality from coronary heart disease (CHD) as compared to both whites and South Asians. Because the male-specific region (MSY) of the human Y chromosome is an obvious candidate for gender-related differences in the development of cardiovascular diseases, we aimed to identify genetic variants of MSY influencing cardiovascular risk profile in different ethnic groups. METHODS AND RESULTS: We genotyped 4 polymorphisms of MSY (HindIII+/-, rs768983 of TBL1Y, rs3212292 of USP9Y, and rs9341273 of UTY genes) in 579 men of different ethnic groups (blacks, South Asians, and whites) from UK and in 301 whites in Italy. We found that the TBL1Y(A) USP9Y(A) haplotype, present only in blacks in whom it represents the most frequent allelic combinations (AA: n=125; all other combinations: n=45), was associated with lower levels of triglycerides (P=0.025) and higher levels of HDL-cholesterol (P=0.005) as compared to the other haplotypes. CONCLUSIONS: The TBL1Y(A) USP9Y(A) haplotype of the Y chromosome, present only in black people of African origin, attributes a favorable lipoprotein pattern, likely to contribute to their reduced susceptibility to coronary heart disease. PMID- 18511698 TI - Femoral plaques confound the association of circulating oxidized low-density lipoprotein with carotid atherosclerosis in a general population aged 35 to 55 years: the Asklepios Study. AB - OBJECTIVE: Reported associations of oxidized low-density lipoprotein (oxLDL) with noninvasive measures of atherosclerosis are inconsistent. In the Asklepios Study cohort of asymptomatic subjects aged 35 to 55 years, we evaluated the relationship of circulating oxLDL with subclinical atherosclerosis in the carotid and femoral arteries. METHODS AND RESULTS: Participants (n=2524, 51.5% females) completed a study questionnaire and underwent a clinical examination, blood analysis of oxLDL (mAb-4E6) and other risk markers, and ultrasound examination of intima-media thickness (IMT) and plaques in the left and right carotid and femoral arteries. oxLDL concentrations were highest in subjects with femoral plaques (n=658). In the group of subjects with carotid plaques (n=476), elevated oxLDL concentrations are related to concomitant femoral plaques detected in 54% of these subjects. Multivariate regression analyses (including anthropometric, hemodynamic, biochemical, and lifestyle variables) showed that femoral plaques are independently related to oxLDL whereas femoral IMT, carotid IMT, or carotid plaques were not independently associated with oxLDL. CONCLUSIONS: Circulating oxLDL is independently associated with femoral plaque and not with carotid artery wall damage. PMID- 18511699 TI - Reevaluation of the role of VEGF-B suggests a restricted role in the revascularization of the ischemic myocardium. AB - OBJECTIVE: The endogenous role of the VEGF family member vascular endothelial growth factor-B (VEGF-B) in pathological angiogenesis remains unclear. METHODS AND RESULTS: We studied the role of VEGF-B in various models of pathological angiogenesis using mice lacking VEGF-B (VEGF-B(-/-)) or overexpressing VEGF B(167). After occlusion of the left coronary artery, VEGF-B deficiency impaired vessel growth in the ischemic myocardium whereas, in wild-type mice, VEGF-B(167) overexpression enhanced revascularization of the infarct and ischemic border zone. By contrast, VEGF-B deficiency did not affect vessel growth in the wounded skin, hypoxic lung, ischemic retina, or ischemic limb. Moreover, VEGF-B(167) overexpression failed to enhance vascular growth in the skin or ischemic limb. CONCLUSIONS: VEGF-B appears to have a relatively restricted angiogenic activity in the ischemic heart. These insights might offer novel therapeutic opportunities. PMID- 18511700 TI - Nonatopic children with multitrigger wheezing have airway pathology comparable to atopic asthma. AB - RATIONALE: Epidemiologic studies have shown that, in atopic children, wheezing is more likely to persist into adulthood, eventually becoming asthma, whereas it appears to resolve by adolescence in nonatopic children. OBJECTIVES: To investigate whether among children with multitrigger wheeze responsive to bronchodilators the airway pathology would be different in nonatopic wheezers, who are often considered nonasthmatic, compared with atopic wheezers, who are more frequently diagnosed as having asthma. METHODS: Bronchial biopsies were obtained from 55 children undergoing bronchoscopy for appropriate clinical indications: 18 nonatopic children with multitrigger wheeze (median age, 5 yr; range, 2-10 yr), 20 atopic children with multitrigger wheeze (medan age, 5 yr; range, 2-15 yr), and 17 control children with no atopy or wheeze (median age, 4; range, 2-14 yr). By histochemistry and immunohistochemistry, we quantified epithelial loss, basement membrane thickness, angiogenesis, inflammatory cells, IL-4(+,) and IL-5(+) cells in subepithelium. MEASUREMENTS AND MAIN RESULTS: Unexpectedly, all pathologic features examined were similar in atopic and nonatopic wheezing children. Compared with control subjects, both nonatopic and atopic wheezing children had increased epithelial loss (P = 0.03 and P = 0.002, respectively), thickened basement membrane (both P < 0.0001), and increased number of vessels (P = 0.003 and P = 0.03, respectively) and eosinophils (P < 0.0001 and P = 0.002, respectively). Moreover, they had increased cytokine expression, which was highly significant for IL-4 (P = 0.002 and P = 0.0001, respectively) and marginal for IL-5 (P = 0.02 and P = 0.08, respectively). CONCLUSIONS: This study shows that the airway pathology typical of asthma is present in nonatopic wheezing children just as in atopic wheezing children. These results suggest that, when multitrigger wheezing responsive to bronchodilators is present, it is associated with pathologic features of asthma even in nonatopic children. PMID- 18511701 TI - Interferon-gamma regulates idiopathic pneumonia syndrome, a Th17+CD4+ T-cell mediated graft-versus-host disease. AB - RATIONALE: Pulmonary complications of hematopoietic stem cell transplantation include infections and graft-versus-host diseases, such as idiopathic pneumonia syndrome (IPS). Conflicting data exist regarding the role of the interferon (IFN) gamma-producing Th1 CD4(+) T-cell subset and IL-17A in IPS. OBJECTIVES: To determine the role of IFN-gamma and IL-17A in the establishment of pulmonary graft-versus-host disease. METHODS: A semiallogeneic murine model based on C57BL/6 x BALB/c as recipients with transplantation of BALB/c RAG2(-/-) bone marrow and transfer of different genetic knockout T cells (T-bet(-/-), IFN-gamma( /-), IFN-gammaR(-/-)) on a BALB/c background. Lung tissue was examined for parenchymal changes and infiltrating cells by histology and fluorescence activated cell sorter analysis. MEASUREMENTS AND MAIN RESULTS: After transfer of semiallogeneic bone marrow together with donor CD4(+) T cells lacking IFN-gamma or T-bet-a T-box transcription factor controlling Th1 commitment-we found severe inflammation in the lungs, but no enhancement in other organs. In contrast, wild type donor CD4(+) T cells mediated minimal inflammation only, and donor CD8(+) T cells were not required for IPS development. Mechanistically, the absence of IFN gamma or IFN-gamma signaling in pulmonary parenchymal cells promoted expansion of IL-17A-producing CD4(+) T cells and local IL-17A release. In vivo depletion of IL 17A reduced disease severity. CONCLUSIONS: One mechanism of IFN-gamma protection against IPS is negative regulation of the expansion of pathogenic IL-17A producing CD4(+) T cells through interaction with the IFN-gamma receptor on the pulmonary parenchymal cell population. PMID- 18511702 TI - Effect of pharmacotherapy on rate of decline of lung function in chronic obstructive pulmonary disease: results from the TORCH study. AB - RATIONALE: Chronic obstructive pulmonary disease (COPD) is characterized by an accelerated decline in lung function. No drug has been shown conclusively to reduce this decline. OBJECTIVES: In a post hoc analysis of the Toward a Revolution in COPD Health (TORCH) study, we investigated the effects of combined salmeterol 50 microg plus fluticasone propionate 500 microg, either component alone or placebo, on the rate of post-bronchodilator FEV(1) decline in patients with moderate or severe COPD. METHODS: A randomized, double-blind, placebo controlled study was conducted from September 2000 to November 2005 in 42 countries. Of 6,112 patients from the efficacy population, 5,343 were included in this analysis. MEASUREMENTS AND MAIN RESULTS: Spirometry was measured every 24 weeks for 3 years. There were 26,539 on-treatment observations. The adjusted rate of decline in FEV(1) was 55 ml/year for placebo, 42 ml/year for salmeterol, 42 ml/year for fluticasone propionate, and 39 ml/year for salmeterol plus fluticasone propionate. Salmeterol plus fluticasone propionate reduced the rate of FEV(1) decline by 16 ml/year compared with placebo (95% confidence interval [CI], 7-25; P < 0.001). The difference was smaller for fluticasone propionate and salmeterol compared with placebo (13 ml/year; 95% CI, 5-22; P = 0.003). Rates of decline were similar among the active treatment arms. FEV(1) declined faster in current smokers and patients with a lower body mass index, and varied between world regions. Patients who exacerbated more frequently had a faster FEV(1) decline. CONCLUSIONS: Pharmacotherapy with salmeterol plus fluticasone propionate, or the components, reduces the rate of decline of FEV(1) in patients with moderate-to-severe COPD, thus slowing disease progression. Clinical trial (GSK Study Code SCO30003) registered with www.clinicaltrials.gov (NCT00268216). PMID- 18511703 TI - Acquired weakness, handgrip strength, and mortality in critically ill patients. AB - RATIONALE: ICU-acquired paresis (ICUAP) is common in survivors of critical illness. There is significant associated morbidity, including prolonged time on the ventilator and longer hospital stay. However, it is unclear whether ICUAP is independently associated with mortality, as sicker patients are more prone and existing studies have not adjusted for this. OBJECTIVES: To test the hypothesis that ICUAP is independently associated with increased mortality. Secondarily, to determine if handgrip dynamometry is a concise measure of global strength and is independently associated with mortality. METHODS: A prospective multicenter cohort study was conducted in intensive care units (ICU) of five academic medical centers. Adults requiring at least 5 days of mechanical ventilation without evidence of preexisting neuromuscular disease were followed until awakening and were then examined for strength. MEASUREMENTS AND MAIN RESULTS: We measured global strength and handgrip dynamometry. The primary outcome was in-hospital mortality and secondary outcomes were hospital and ICU-free days, ICU readmission, and recurrent respiratory failure. Subjects with ICUAP (average MRC score of < 4) had longer hospital stays and required mechanical ventilation longer. Handgrip strength was lower in subjects with ICUAP and had good test performance for diagnosing ICUAP. After adjustment for severity of illness, ICUAP was independently associated with hospital mortality (odds ratio [OR], 7.8; 95% confidence interval [CI], 2.4-25.3; P = 0.001). Separately, handgrip strength was independently associated with hospital mortality (OR, 4.5; 95% CI, 1.5-13.6; P = 0.007). CONCLUSIONS: ICUAP is independently associated with increased hospital mortality. Handgrip strength is also independently associated with poor hospital outcome and may serve as a simple test to identify ICUAP. Clinical trial registered with www.clinicaltrials.gov (NCT00106665). PMID- 18511704 TI - Blunted hypoxic pulmonary vasoconstriction in experimental neonatal chronic lung disease. AB - RATIONALE: Neonatal chronic lung disease (CLD), caused by prolonged mechanical ventilation (MV) with O(2)-rich gas, is the most common cause of long-term hospitalization and recurrent respiratory illness in extremely premature infants. Recurrent episodes of hypoxemia and associated ventilator adjustments often lead to worsening CLD. The mechanism that causes these hypoxemic episodes is unknown. Hypoxic pulmonary vasoconstriction (HPV), which is partially controlled by O(2) sensitive voltage-gated potassium (K(v)) channels, is an important adaptive response to local hypoxia that helps to match perfusion and ventilation in the lung. OBJECTIVES: To test the hypothesis that chronic lung injury (CLI) impairs HPV. METHODS: We studied preterm lambs that had MV with O(2)-rich gas for 3 weeks and newborn rats that breathed 95%-O(2) for 2 weeks, both of which resulted in airspace enlargement and pulmonary vascular changes consistent with CLD. MEASUREMENTS AND MAIN RESULTS: HPV was attenuated in preterm lambs with CLI after 2 weeks of MV and in newborn rats with CLI after 2 weeks of hyperoxia. HPV and constriction to the K(v)1.x-specific inhibitor, correolide, were preferentially blunted in excised distal pulmonary arteries (dPAs) from hyperoxic rats, whose dPAs exhibited decreased K(v)1.5 and K(v)2.1 mRNA and K(+) current. Intrapulmonary gene transfer of K(v)1.5, encoding the ion channel that is thought to trigger HPV, increased O(2)-sensitive K(+) current in cultured smooth muscle cells from rat dPAs, and restored HPV in hyperoxic rats. CONCLUSIONS: Reduced expression/activity of O(2)-sensitive K(v) channels in dPAs contributes to blunted HPV observed in neonatal CLD. PMID- 18511705 TI - Increased p53 level, Bax/Bcl-x(L) ratio, and TRAIL receptor expression in human emphysema. AB - RATIONALE: Emphysema is mainly known for the complex inflammatory processes associated with its development. In addition to lung inflammation, it is now accepted that increased alveolar cell apoptosis is also part of emphysema pathophysiology. However, little is known about the mechanisms involved in alveolar apoptosis. We postulate that oxidative stress and proinflammatory cytokines could lead to p53 accumulation, Bax/Bcl-x(L) ratio elevation, and higher tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptor levels in the emphysematous lung. OBJECTIVES: To evaluate the expression of p53, Bax, Bcl-x(L), TRAIL, and TRAIL receptors in lung parenchyma from nonemphysematous nonsmokers and smokers and emphysematous smokers and ex-smokers and to determine whether H2O2 and/or TNF can modulate the expression of these apoptotic proteins. METHODS: p53, Bax, Bcl-x(L), and TRAIL receptor protein levels in lung parenchyma were measured by Western blot, and TRAIL mRNA levels were measured by real-time polymerase chain reaction. Changes in TRAIL receptor, Bax, Bcl-x(L), and p53 protein levels after in vitro H2O2 and/or TNF stimulation of A549 cells were also assessed by Western blot. MEASUREMENTS AND MAIN RESULTS: The p53 protein levels, the Bax/Bcl-x(L) ratio, and TRAIL receptors 1, 2, and 3 protein levels were significantly higher in subjects with emphysema. Moreover, they were also increased after H2O2 and TNF treatments of A549 cells. CONCLUSIONS: These findings suggest that oxidative stress and proinflammatory cytokines may be involved in the elevation of p53 levels, the Bax/Bcl-x(L) ratio, and TRAIL receptor levels, new mechanisms that may be implicated in the increased alveolar cell apoptosis that occurs in emphysema. PMID- 18511706 TI - Influence of multidrug resistance on tuberculosis treatment outcomes with standardized regimens. AB - RATIONALE: Multidrug-resistant tuberculosis (TB) poses a major challenge to global TB control. We analyzed the association between estimated prevalence of initial or acquired MDR-TB, and treatment outcomes reported nationally. OBJECTIVES: We analyzed the estimated prevalence of initial or acquired MDR-TB and treatment outcomes reported nationally. METHODS: Countries were analyzed if multidrug resistance prevalence estimates were available, and if they reported outcomes for more than 250 cases treated using standardized regimens in 2003 and/or 2004. Data sources were the World Health Organization for treatment regimens, prevalence of initial multidrug resistance, and reported cases and treatment outcomes in 2003 and 2004; the Joint United Nations Programme on HIV/AIDS for HIV seroprevalence; and the World Bank for income per capita. The adjusted impact of initial multidrug resistance on initial and retreatment outcomes was estimated with weighted multivariate linear regression. MEASUREMENTS AND MAIN RESULTS: Among countries using one of two standardized initial regimens, failure rates averaged 5.0%, and relapse rates averaged 12.8% in the 20 countries where prevalence of initial multidrug resistance exceeded 3%, compared with an average of 1.6% (P < 0.0001) and 8.1% (P = 0.0002), respectively, in 83 countries where initial multidrug resistance prevalence was less than 3%. In 92 countries using one standardized retreatment regimen, failure rates were 2.7%, 3.8%, 6.2%, and 8.1% in quartiles of increasing prevalence of acquired multidrug resistance (P < 0.0001). When stratified by initial multidrug resistance prevalence, initial and retreatment outcomes in the 79 countries using the 6-month rifampin initial regimen were not significantly different from the 24 countries using the 2-month rifampin initial regimen. CONCLUSIONS: Currently recommended standardized TB initial and retreatment regimens should be reevaluated in countries where prevalence of initial multidrug resistance exceeds 3%, in view of poor treatment outcomes. PMID- 18511707 TI - Interleukin-8 as a stratification tool for interventional trials involving pediatric septic shock. AB - RATIONALE: Interventional clinical trials involving children with septic shock would benefit from an efficient preenrollment stratification strategy. OBJECTIVES: To test the predictive value of interleukin (IL)-8 for 28-day mortality in pediatric septic shock. METHODS: A training data set (n = 40) identified a serum IL-8 of greater than 220 pg/ml as having a 75% sensitivity and specificity for predicting 28-day mortality. This cutoff was then subjected to a series of validation steps. MEASUREMENTS AND MAIN RESULTS: Subjects were drawn from two large, independent pediatric septic shock databases. Prospective application of the IL-8 cutoff to validation data set 1 (n = 139) demonstrated 78% sensitivity and 64% specificity for 28-day mortality. A serum IL-8 level of 220 pg/ml or less, however, had a negative predictive value for 28-day mortality of 95% in validation data set 1, which was subsequently applied to an independently generated data set of children with septic shock (validation set 2, n = 193). A serum IL-8 level of 220 pg/ml or less had a negative predictive value for 28-day mortality of 94% when applied to validation set 2. CONCLUSIONS: A serum IL-8 level of 220 pg/ml or less, obtained within 24 hours of admission, predicts a high likelihood of survival in children with septic shock. We propose that IL-8 can be used to exclude such patients from interventional clinical trials and ultimately derive a study population with a more favorable risk to benefit ratio when subjected to a study agent. PMID- 18511708 TI - mTOR mediates survival signals in malignant mesothelioma grown as tumor fragment spheroids. AB - Solid tumors such as mesothelioma exhibit a stubborn resistance to apoptosis that may derive from survival pathways, such as PI3K/Akt/mTOR, that are activated in many tumors, including mesothelioma. To address the role of PI3K/Akt/mTOR, we used a novel approach to study mesothelioma ex vivo as tumor fragment spheroids. Freshly resected mesothelioma tissue from 15 different patients was grown in vitro as 1- to 2-mm-diameter fragments, exposed to apoptotic agents for 48 hours with or without PI3K/Akt/mTOR inhibitors, and doubly stained for cytokeratin and cleaved caspase 3 to identify apoptotic mesothelioma cells. Mesothelioma cells within the tumor spheroids exhibited striking resistance to apoptotic agents such as TRAIL plus gemcitabine that were highly effective against monolayers. In a majority of tumors (67%; 10 of 15), apoptotic resistance could be reduced by more than 50% by rapamycin, an mTOR inhibitor, but not by LY294002, a PI3K inhibitor. Responsiveness to rapamycin correlated with staining for the mTOR target, p-S6K, in the original tumor, but not for p-Akt. As confirmation of the role of mTOR, siRNA knockdown of S6K reproduced the effect of rapamycin in three rapamycin responsive tumors. Finally, in 37 mesotheliomas on tissue microarray, p-S6K correlated only weakly with p-Akt, suggesting the existence of Akt-independent regulation of mTOR. We propose that mTOR mediates survival signals in many mesothelioma tumors. Inhibition of mTOR may provide a nontoxic adjunct to therapy directed against malignant mesothelioma, especially in those with high baseline expression of p-S6K. PMID- 18511709 TI - Mice lacking 12/15-lipoxygenase have attenuated airway allergic inflammation and remodeling. AB - Arachidonate 15-lipoxygenase (LO)-1 has been implicated in allergic inflammation and asthma. The overall effect of 15-LO in allergic inflammation in vivo is, however, unclear. This study investigates systemic allergen sensitization and local allergic airway inflammation and remodeling in mice lacking the murine 12/15-LO, the ortholog to human 15-LO-1. Upon systemic sensitization with intraperitoneal ovalbumin, 12/15-LO-/- mice produced elevated levels of allergen specific immunoglobulin E compared with wild-type (Wt) controls. However, when challenged with repeated aerosolized allergen, sensitized 12/15-LO-/- mice had an impaired development of airway allergic inflammation compared with Wt controls, as indicated by reduced bronchoalveolar lavage fluid leukocytes (eosinophils, lymphocytes, macrophages) and Th2 cytokines (IL-4, IL-5, IL-13), as well as tissue eosinophils. Allergen-induced airway epithelial proliferation was also significantly attenuated in 12/15-LO-/- mice, whereas goblet cell hyperplasia was unaffected. However, 12/15-LO-/- mice had significantly reduced luminal mucus secretions compared with Wt controls. The repeated allergen challenges resulted in a dramatic increase of alpha-smooth muscle actin-positive alveolar cells in the peripheral airways, a phenomenon that was significantly less developed in 12/15-LO-/- mice. In conclusion, our data suggest that 12/15-LO-/- mice, although having a fully developed systemic sensitization, did not establish a fully developed allergic airway inflammation and associated manifestations of central and peripheral airway remodeling. These data suggest that 12/15-LO-derived metabolites play an important pathophysiologic role in allergen-induced inflammation and remodeling. Hence, pharmacologic targeting of the human 15-LO-1 may represent an attractive therapeutic strategy to control inflammation and remodeling in asthma. PMID- 18511710 TI - Lead poisoning in immigrant children in the United States. PMID- 18511711 TI - James Fries: healthy aging pioneer. PMID- 18511712 TI - Subdividing the age group of 85 years and older to improve US disease reporting. AB - The standard terminal age category in disease reporting in the United States has been 85 years and older since the 1940s, but the dramatically increasing share of the US population reaching this age has rendered the single category inadequate for surveillance, research, and analysis. Important age-specific variations in mortality among the oldest old are masked by the continued use of this category. Greater specificity in age-specific data for the oldest old would aid in disease surveillance and etiologic research and broaden awareness and understanding of human longevity. PMID- 18511713 TI - Controlling personal health decisions for the oldest old. PMID- 18511714 TI - Chronic conditions and mortality among the oldest old. AB - OBJECTIVES: We sought to determine whether chronic conditions and functional limitations are equally predictive of mortality among older adults. METHODS: Participants in the 1998 wave of the Health and Retirement Study (N=19430) were divided into groups by decades of age, and their vital status in 2004 was determined. We used multivariate Cox regression to determine the ability of chronic conditions and functional limitations to predict mortality. RESULTS: As age increased, the ability of chronic conditions to predict mortality declined rapidly, whereas the ability of functional limitations to predict mortality declined more slowly. In younger participants (aged 50-59 years), chronic conditions were stronger predictors of death than were functional limitations (Harrell C statistic 0.78 vs. 0.73; P=.001). In older participants (aged 90-99 years), functional limitations were stronger predictors of death than were chronic conditions (Harrell C statistic 0.67 vs. 0.61; P=.004). CONCLUSIONS: The importance of chronic conditions as a predictor of death declined rapidly with increasing age. Therefore, risk-adjustment models that only consider comorbidities when comparing mortality rates across providers may be inadequate for adults older than 80 years. PMID- 18511715 TI - Influence of musculoskeletal conditions on oral health among older adults. AB - Both musculoskeletal disorders and diseases of the oral cavity are common and potentially serious problems among older persons, yet little attention has been given to the links between them. Several musculoskeletal diseases, including osteoporosis, Paget's disease, and arthritic disorders, may directly involve the oral cavity and contiguous structures. Drugs used to treat musculoskeletal diseases, including corticosteroids and bisphosphonates, increase the risk of suppression of the immune system and osteonecrosis of the jaw, respectively. Many people with disabling osteoarthritis, rheumatoid arthritis, and other conditions have difficulty practicing good oral hygiene and traveling to dental offices for professional help. Various inexpensive measures can help such individuals, including education of their caregivers and provision of antimicrobial mouthwashes and special toothbrushes. PMID- 18511716 TI - Temporal trends in self-reported functional limitations and physical disability among the community-dwelling elderly population: the Framingham heart study. AB - OBJECTIVES: We sought to determine change in the prevalence of functional limitations and physical disability among the community-dwelling elderly population across 3 decades. METHODS: We studied original participants of the Framingham Heart Study, aged 79 to 88 years, at examination 15 (1977-1979; 177 women, 103 men), examination 20 (1988-1990; 159 women, 98 men) and examination 25 (1997-1999; 174 women, 119 men). Self-reported functional limitation was defined using the Nagi scale, and physical disability was defined using the Rosow-Breslau and Katz scales. RESULTS: Functional limitations declined across examinations from 74.6% to 60.5% to 37.9% (P < .001) among women and from 54.2% to 37.8% to 27.8% (P<.001) among men. Physical disability declined from 74.5% to 48.5% to 34.6% (P < .001) among women and 42.3% to 33.3% to 22.8% (P = .009) among men. Among women, improvements in functional limitations (P = .05) were greater from examination 20 to 25, whereas for physical disability (P=.02), improvements were greater from examination 15 to 20. Improvements in function were constant across the 3 examinations in men. CONCLUSIONS: Among community-dwelling elders, the prevalence of functional limitations and physical disability declined significantly in both women and men from the 1970s to the 1990s. This may in part be due to improvements in technological devices used to maintain independence. Further work is needed to identify the underlining causes of the decline so preventative measures can be established that promote independence for the elderly population. PMID- 18511717 TI - Exploring potential pathways between parity and tooth loss among American women. AB - OBJECTIVES: We assessed the impact of parity on tooth loss among American women and examined mediators of this relationship. METHODS: The study sample comprised 2635 White and Black non-Hispanic women who had taken part in the third National Health and Nutrition Examination Survey. We examined the relationship between parity and tooth loss, by age and by socioeconomic position, and tested a theoretical model focusing on direct and indirect influences of parity on dental disease. Robust regression techniques were used to generate path coefficients. RESULTS: Although parity was associated with tooth loss, the relationship was not moderated through dental care, psychosocial factors, or dental health-damaging behaviors. CONCLUSIONS: Parity is related to tooth loss among American women, but the mechanisms of the association remain undefined. Further investigation is warranted to determine whether disparities in dental health among women who have been pregnant are caused by differences in parity or to physiological and societal changes (e.g., factors related to pregnant women's access to care) paralleling reproductive choices. PMID- 18511718 TI - Associations of acculturation and socioeconomic status with subclinical cardiovascular disease in the multi-ethnic study of atherosclerosis. AB - OBJECTIVES: We assessed whether markers of acculturation (birthplace and number of US generations) and socioeconomic status (SES) are associated with markers of subclinical cardiovascular disease-carotid artery plaque, internal carotid intima media thickness, and albuminuria-in 4 racial/ethnic groups. METHODS: With data from the Multi-Ethnic Study of Atherosclerosis (n = 6716 participants aged 45-84 years) and race-specific binomial regression models, we computed prevalence ratios adjusted for demographics and traditional cardiovascular risk factors. RESULTS: The adjusted US- to foreign-born prevalence ratio for carotid plaque was 1.20 (99% confidence interval [CI] = 0.97, 1.39) among Whites, 1.91 (99% CI = 0.94, 2.94) among Chinese, 1.62 (99% CI = 1.28, 2.06) among Blacks, and 1.23 (99% CI = 1.15, 1.31) among Hispanics. Greater carotid plaque prevalence was found among Whites, Blacks, and Hispanics with a greater number of generations with US residence (P < .001) and among Whites with less education and among Blacks with lower incomes. Similar associations were observed with intima-media thickness. There was also evidence of an inverse association between albuminuria and SES among Whites and Hispanics. CONCLUSIONS: Greater US acculturation and lower SES were associated with a higher prevalence of carotid plaque and greater intima media thickness but not with albuminuria. Maintenance of healthful habits among recent immigrants should be encouraged. PMID- 18511719 TI - Multivitamin-mineral supplements in the Older Americans Act Nutrition Program: not a one-size-fits-all quick fix. AB - We challenge the suggestion of Congress that the Older Americans Act (OAA) Nutrition Program should provide multivitamin-mineral supplements (MVMs) in addition to meals. MVMs are not a quick fix for poor diets. They do not contain calories, protein, essential fatty acids, or fiber, nor do they adequately address nutritional gaps of some vitamins and minerals. Older adults with chronic health conditions who take multiple medications are at greater risk than the general healthy population for nutrient-drug interactions and toxicity. The OAA Nutrition Program is not an appropriate venue to indiscriminately distribute MVMs, because there is insufficient evidence of their benefits and safety. The program's limited funds and efforts should instead be directed to nutrient-dense healthy meals, quality food service, and greater accessibility to individualized nutrition services. PMID- 18511720 TI - The risks, costs, and benefits of possible future global policies for managing polioviruses. AB - OBJECTIVES: We assessed the costs, risks, and benefits of possible future major policy decisions on vaccination, surveillance, response plans, and containment following global eradication of wild polioviruses. METHODS: We developed a decision analytic model to estimate the incremental cost-effectiveness ratios and net benefits of risk management options for polio for the 20-year period and stratified the world according to income level to capture important variability between nations. RESULTS: For low-, lower-middle-, and upper-middle-income groups currently using oral poliovirus vaccine (OPV), we found that after successful eradication of wild polioviruses, OPV cessation would save both costs and lives when compared with continued use of OPV without supplemental immunization activities. We found cost-effectiveness ratios for switching from OPV to inactivated poliovirus vaccine to be higher (i.e., less desirable) than other health investment opportunities, depending on the actual inactivated poliovirus vaccine costs and assumptions about whether supplemental immunization activities with OPV would continue. CONCLUSIONS: Eradication promises billions of dollars of net benefits, although global health policy leaders face difficult choices about future policies. Until successful eradication and coordination of posteradication policies, health authorities should continue routine polio vaccination and supplemental immunization activities. PMID- 18511721 TI - The secret and soul of Marlboro: Phillip Morris and the origins, spread, and denial of nicotine freebasing. AB - Philip Morris and other tobacco companies have been using ammonia in their manufacturing for more than half a century, and for a variety of purposes: to highlight certain flavors, to expand or "puff up" the volume of tobacco, to prepare reconstituted tobacco sheet ("recon"), to denicotinize (reduce the amount of nicotine in) tobacco, and to remove carcinogens. By the early 1960s, however, Philip Morris had also begun using ammonia to "freebase" the nicotine in cigarette smoke, creating low-yield (reduced-tar or -nicotine) cigarettes that still had the nicotine kick necessary to keep customers "satisfied" (i.e., addicted). We show that Philip Morris discovered the virtues of freebasing while analyzing the impact of the ammoniated recon used in Marlboro cigarettes. We also show how Marlboro's commercial success catalyzed efforts by the rest of the tobacco industry to discover its "secret," eventually identified as ammonia technology, and how Philip Morris later exploited the myriad uses of ammonia (e.g., for flavoring and expanding tobacco volume) to defend itself against charges of manipulating the nicotine deliveries of its cigarettes. PMID- 18511722 TI - Prevalence of chronic diseases and multimorbidity among the elderly population in Sweden. AB - We explored the role of age, gender, and socioeconomic status in the occurrence of chronic diseases and multimorbidity in 1099 elderly participants in the Kungsholmen Project. Cardiovascular and mental diseases were the most common chronic disorders. Of the participants, 55% had multimorbidity. Advanced age, female gender, and lower education were independently associated with a more than 50% increased risk for multimorbidity. Multimorbidity is the most common clinical picture of the elderly and may be increased by unhealthy behaviors linked to education. PMID- 18511723 TI - Translation of an effective tai chi intervention into a community-based falls prevention program. AB - Tai chi--moving for better balance, a falls-prevention program developed from a randomized controlled trial for community-based use, was evaluated with the re aim framework in 6 community centers. The program had a 100% adoption rate and 87% reach into the target older adult population. All centers implemented the intervention with good fidelity, and participants showed significant improvements in health-related outcome measures. This evidence-based tai chi program is practical to disseminate and can be effectively implemented and maintained in community settings. PMID- 18511724 TI - Regular vigorous physical activity and disability development in healthy overweight and normal-weight seniors: a 13-year study. AB - OBJECTIVES: We examined the relationship of regular exercise and body weight to disability among healthy seniors. METHODS: We assessed body mass index (BMI) and vigorous exercise yearly (1989-2002) in 805 participants aged 50 to 72 years at enrollment. We studied 4 groups: normal-weight active (BMI< 25 kg/m(2); exercise> 60 min/wk); normal-weight inactive (exerciseor= 25 kg/m(2)); and overweight inactive. Disability was measured with the Health Assessment Questionnaire (0-3; 0= no difficulty, 3= unable to do). We used multivariable analysis of covariance to determine group differences in disability scores after adjustment for determinants of disability. RESULTS: The cohort was 72% men and 96% White, with a mean age of 65.2 years. After 13 years, overweight active participants had significantly less disability than did overweight inactive (0.14 vs 0.19; P= .001) and normal-weight inactive (0.22; P= .03) participants. Similar differences were found between normal-weight active (0.11) and normal-weight inactive participants (P< .001). CONCLUSIONS: Being physically active mitigated development of disability in these seniors, largely independent of BMI. Public health efforts that promote physically active lifestyles among seniors may be more successful than those that emphasize body weight in the prevention of functional decline. PMID- 18511725 TI - Project VIVA: a multilevel community-based intervention to increase influenza vaccination rates among hard-to-reach populations in New York City. AB - OBJECTIVES: We sought to determine whether the work of a community-based participatory research partnership increased interest in influenza vaccination among hard-to-reach individuals in urban settings. METHODS: A partnership of researchers and community members carried out interventions for increasing acceptance of influenza vaccination in disadvantaged urban neighborhoods, focusing on hard-to-reach populations (e.g., substance abusers, immigrants, elderly, sex workers, and homeless persons) in East Harlem and the Bronx in New York City. Activities targeted the individual, community organization, and neighborhood levels and included dissemination of information, presentations at meetings, and provision of street-based and door-to-door vaccination during 2 influenza vaccine seasons. Participants were recruited via multiple modalities. Multivariable analyses were performed to compare interest in receiving vaccination pre- and postintervention. RESULTS: There was increased interest in receiving the influenza vaccine postintervention (P<.01). Being a member of a hard-to-reach population (P=.03), having ever received an influenza vaccine (P<.01), and being in a priority group for vaccination (P<.01) were also associated with greater interest in receiving the vaccine. CONCLUSIONS: Targeting underserved neighborhoods through a multilevel community-based participatory research intervention significantly increased interest in influenza vaccination, particularly among hard-to-reach populations. Such interventions hold promise for increasing vaccination rates annually and in pandemic situations. PMID- 18511726 TI - Institutionalization of older adults after the death of a spouse. AB - OBJECTIVES: We investigated the risk of entering long-term institutional care after the death of a spouse in relation to the duration of widowhood among older Finnish men and women. We also examined whether high levels of education or household income buffered the effects of bereavement on institutionalization. METHODS: We used linked register-based data on Finnish adults 65 years or older who were living with a spouse at the beginning of the study period (n=140902) and were followed from January 1998 to December 2002. RESULTS: The excess risk of institutionalization was highest during the first month following a spouse's death compared with still living with a spouse (adjusted hazard ratio=3.31 for men, 3.62 for women). This risk decreased over time among both men and women. The relative effect of the duration of widowhood on institutionalization did not significantly vary according to the level of education or income. CONCLUSIONS: Risk of institutionalization is particularly high immediately after the death of a spouse, demonstrating the importance of loss of social and instrumental support. PMID- 18511727 TI - Detection of very early oral cancers in Puerto Rico. AB - We evaluated a possible disparity in the detection of very early oral cancers in Puerto Rico relative to the United States. The percentage of in situ (noninvasive) cases among all oral cancer cases was calculated separately for Puerto Rico and the United States using population-based cancer registry data (1992-2001). In situ cancers constituted 1.2% of oral cancer cases in Puerto Rico and 3.4% in the United States (P<.001). These findings suggest a disparity in very early oral cancer detection in Puerto Rico compared with the United States. PMID- 18511728 TI - Gender and race/ethnicity differences in lead dose biomarkers. AB - OBJECTIVES: We sought to identify predictors of lead concentrations in the blood, tibias, and patellae of older adults and to describe differences by gender, race/ethnicity, and other factors that can influence lead toxicokinetics and, thus modify health effects. METHODS: Participants aged 50 to 70 years (N=1140) were randomly identified from selected neighborhoods in Baltimore, Maryland. We measured lead concentrations by anodic stripping voltammetry (in blood) and (109)Cd-induced K-shell x-ray fluorescence (in bone). We used multiple linear regression to identify predictors of lead concentrations. RESULTS: Mean (SD) lead concentrations in blood, tibias, and patellae were 3.5 (2.4) mug/dL, 18.9 (12.5) mug/g, and 6.8 (18.1) mug/g, respectively. Tibia concentrations were 29% higher in African Americans than in Whites (P < .01). We observed effect modification by race/ethnicity on the association of gender and physical activity to blood lead concentrations and by gender on the association of age to tibia lead concentrations. Patella lead concentrations differed by gender; apolipoprotein E genotype modified this relation. CONCLUSIONS: African Americans evidenced a prominent disparity in lifetime lead dose. Women may be at higher risk of release of lead from bone and consequent health effects because of increased bone demineralization with aging. PMID- 18511729 TI - Medicaid prescription formulary restrictions and arthritis treatment costs. AB - OBJECTIVES: We used the Arizona Medicaid program as a model to examine the consequences of the relative restrictiveness of nonsteroidal anti-inflammatory drug (NSAID)-preferred drug lists on health care use and costs for Medicaid enrollees with arthritis. METHODS: In a retrospective, cross-sectional study of Medicaid enrollees with rheumatoid arthritis or osteoarthritis, we used data from the Arizona HealthQuery database and generalized linear regression models to estimate the effect of the restrictiveness of formularies on the association between number of NSAID drugs covered and the number of emergency department visits, ambulatory physician visits, hospital stays, and total health expenditures. RESULTS: For plans with NSAID formularies that were more restrictive, enrollees with rheumatoid arthritis experienced 22% fewer ambulatory visits and 29% more hospitalizations, and enrollees with osteoarthritis experienced 38% fewer ambulatory visits and 52% more hospitalizations. These plans spent an additional $935 for medical care and prescription drugs annually per enrollee with rheumatoid arthritis. CONCLUSIONS: Formularies that are more restrictive significantly change the patterns of health care and prescription drug use and may have unintended consequences in terms of more frequent and, for those with rheumatoid arthritis, more expensive medical care. PMID- 18511730 TI - Socioeconomic disadvantage and kidney disease in the United States, Australia, and Thailand. AB - OBJECTIVES: We sought to determine whether an elevated burden of chronic kidney disease is found among disadvantaged groups living in the United States, Australia, and Thailand. METHODS: We used data on participants 35 years or older for whom a valid serum creatinine measurement was available from studies in the United States, Thailand, and Australia. We used logistic regression to analyze the association of income, education, and employment with the prevalence of chronic kidney disease (estimated glomerular filtration rate<60 mL/min/1.73 m(2)). RESULTS: Age- and gender-adjusted odds of having chronic kidney disease were increased 86% for US Whites in the lowest income quartile versus the highest quartile (odds ratio [OR] = 1.86; 95% confidence interval [CI] = 1.27, 2.72). Odds were increased 2 times and 6 times, respectively, among unemployed (not retired) versus employed non-Hispanic Black and Mexican American participants (OR=2.89; 95% CI=1.53, 5.46; OR=6.62; 95% CI=1.94, 22.64. respectively). Similar associations were not evident for the Australian or Thai populations. CONCLUSIONS: Higher kidney disease prevalence among financially disadvantaged groups in the United States should be considered when chronic kidney disease prevention and management strategies are created. This approach is less likely to be of benefit to the Australian and Thai populations. PMID- 18511731 TI - Social network, cognitive function, and dementia incidence among elderly women. AB - OBJECTIVES: We examined whether social networks had a protective association with incidence of dementia among elderly women. METHODS: We prospectively studied 2249 members of a health maintenance organization who were 78 years or older, were classified as free of dementia in 2001, and had completed at least 1 follow-up interview in 2002 through 2005. We used the Telephone Interview for Cognitive Status-modified, the Telephone Dementia Questionnaire, and medical record review to assess cognitive status. We used the Lubben Social Network Scale-6 to assess social network. We estimated hazard ratios for incident dementia with Cox proportional hazards models, adjusting for age at entry, education, hormone use, cognitive status scores, and health conditions. RESULTS: We identified 268 incident cases of dementia during follow-up. Compared with women with smaller social networks, the adjusted hazard ratio for incident dementia in women with larger social networks was 0.74 (95% confidence interval=0.57, 0.97). CONCLUSIONS: Our findings suggest that larger social networks have a protective influence on cognitive function among elderly women. Future studies should explore which aspects of social networks are associated with dementia risk and maintenance of cognitive health. PMID- 18511732 TI - Perceived discrimination and mortality in a population-based study of older adults. AB - OBJECTIVES: We examined the relation of individual-level perceived discrimination to mortality in a biracial, population-based sample. METHODS: Participants were 4154 older adults from the Chicago Health and Aging Project who underwent up to 2 interviews over 4.5 years. Perceived discrimination was measured at baseline, and vital status was obtained at each follow-up and verified through the National Death Index. RESULTS: During follow-up, 1166 deaths occurred. Participants reporting more perceived discrimination had a higher relative risk of death (hazard ratio [HR]= 1.05; 95% confidence interval [CI]=1.01, 1.09). This association was independent of differences in negative affect or chronic illness and appeared to be stronger among Whites than among Blacks (Whites: HR=1.12; 95% CI=1.04, 1.20; Blacks: HR=1.03; 95% CI=0.99, 1.07). Secondary analyses revealed that the relation to mortality was related to discriminatory experiences of a more demeaning nature and that racial differences were no longer significant when the sample was restricted to respondents interviewed by someone of the same race. CONCLUSIONS: Perceived discrimination was associated with increased mortality risk in a general population of older adults. The results suggest that subjective experience of interpersonal mistreatment is toxic in old age. This study adds to a growing literature documenting discrimination as an important social determinant of health. PMID- 18511733 TI - The effect of widowhood on mortality by the causes of death of both spouses. AB - OBJECTIVES: We investigated the effect of spousal bereavement on mortality to document cause-specific bereavement effects by the causes of death of both the predecedent spouse and the bereaved partner. METHODS: We obtained data from a nationally representative cohort of 373 189 elderly married couples in the United States who were followed from 1993 to 2002. We used competing risk and Cox models in our analyses. RESULTS: For both men and women, the death of a predecedent spouse from almost all causes, including various cancers, infections, and cardiovascular diseases, increased the all-cause mortality of the bereaved partner to varying degrees. Moreover, the death of a predecedent spouse from any cause increased the survivor's cause-specific mortality for almost all causes, including cancers, infections, and cardiovascular diseases, to varying degrees. CONCLUSIONS: The effect of widowhood on mortality varies substantially by the causes of death of both spouses, suggesting that the widowhood effect is not restricted to one aspect of human biology. Future research should examine the specific mechanisms of the widowhood effect and identify opportunities for health interventions. PMID- 18511734 TI - Prospective change in health-related quality of life and subsequent mortality among middle-aged and older women. AB - OBJECTIVES: We sought to determine prospective changes in health-related quality of life (HRQoL) measures and subsequent mortality in middle-aged and older women. METHODS: We obtained data from 40 337 healthy women from the Nurses' Health Study aged 46 to 71 years in 1992. We used Cox proportional hazards regression to evaluate associations of changes in self-assessed physical and mental component summary (PCS and MCS) scores from the Short Form 36 Health Survey between 1992 and 1996 and between 1996 and 2000, with all-cause mortality through 2004. RESULTS: Women with low HRQoL (PCS and MCS scores) and the greatest HRQoL declines had higher mortality than did women with stable scores. Change in PCS score predicted mortality across the range of 4-year change: severe decline (relative risk [RR] = 3.32; 95% confidence interval [CI] = 2.45, 4.50), moderate decline (RR = 1.44; 95% CI = 1.16, 1.79), slight decline (RR = 1.35; 95% CI = 1.12, 1.63), no change (reference category), improvement (RR = 0.72; 95% CI = 0.56, 0.91; continuous P < .001). MCS score results were similar. Score increases were associated with lifestyle improvements, especially increased physical activity. CONCLUSIONS: Observed associations demonstrate the predictive validity of changes in self-assessed HRQoL for subsequent mortality in healthy populations. Future research should examine determinants of patterns of change. PMID- 18511735 TI - Predictors of longevity: evidence from the oldest old in China. AB - OBJECTIVES: We investigated the characteristics of the oldest old in China and examined whether the factors associated with longevity varied with advanced age. METHODS: Drawing from the largest nationally representative longitudinal sample of oldest-old adults, we stratified descriptive statistics separately by gender and urban-rural residence and then used ordered logit models to examine the multivariate factors associated with increasing age-group membership. RESULTS: Differing combinations of demographic, social, physical, and behavioral factors were significantly related to surviving into later ages for men and women in urban and rural areas. With the exception of rural women, psychological disposition was not associated with increased longevity. Gender differences were generally smaller in urban areas than in rural areas, and urban-rural differences were more pronounced among women than among men. CONCLUSIONS: Findings from the oldest-old population in China challenge many of the established relations in the health-inequality literature. Future research should examine why the oldest old are an exceptional group of physically, socially, and demographically heterogeneous individuals who exhibit healthy longevity beyond the average life span. PMID- 18511736 TI - Effects of social integration on preserving memory function in a nationally representative US elderly population. AB - OBJECTIVES: We tested whether social integration protects against memory loss and other cognitive disorders in late life in a nationally representative US sample of elderly adults, whether effects were stronger among disadvantaged individuals, and whether earlier cognitive losses explained the association (reverse causation). METHODS: Using data from the Health and Retirement Study (N = 16,638), we examined whether social integration predicted memory change over 6 years. Memory was measured by immediate and delayed recall of a 10-word list. Social integration was assessed by marital status, volunteer activity, and frequency of contact with children, parents, and neighbors. We examined growth curve models for the whole sample and within subgroups. RESULTS: The mean memory score declined from 11.0 in 1998 to 10.0 in 2004. Higher baseline social integration predicted slower memory decline in fully adjusted models (P<.01). Memory among the least integrated declined at twice the rate as among the most integrated. This association was largest for respondents with fewer than 12 years of education. There was no evidence of reverse causation. CONCLUSIONS: Our study provides evidence that social integration delays memory loss among elderly Americans. Future research should focus on identifying the specific aspects of social integration most important for preserving memory. PMID- 18511737 TI - Relation between the level of American Indian and Alaska Native diabetes education program services and quality-of-care indicators. AB - OBJECTIVES: We examined the relation between the level of diabetes education program services in the Indian Health Service (IHS) and indicators of the quality of diabetes care to determine if more-comprehensive diabetes services were associated with better quality of diabetes care. METHODS: In this cross-sectional study, we used the IHS Integrated Diabetes Education Recognition Program to rank program services into 1 of 3 levels of comprehensiveness, ranging from lowest (developmental) to highest (integrated). We compared quality-of-care indicators among programs of differing levels with the 2001 IHS Diabetes Care and Outcomes Audit. Quality indicators included patients having recommended yearly examinations, education, and laboratory tests and achieving recommended levels of intermediate outcomes of care. RESULTS: Most of the 86 participating programs were classified at or below the developmental level; only 9 programs (11%) were ranked at higher levels. After adjusting for patient characteristics, program factors, and correlation of patients within programs, we associated programs that were more comprehensive with higher completion rates of yearly lipid and hemoglobin A1C tests (P < .05). CONCLUSIONS: System-wide improvements in diabetes education are associated with better diabetes care. The results can help inform the development of diabetes education programs. PMID- 18511738 TI - Mental health status among rural women of reproductive age: findings from the Central Pennsylvania Women's Health Study. AB - OBJECTIVES: We sought to examine variables associated with mental health among rural women of reproductive age, with particular attention given to rural area type and farm residence. METHODS: We analyzed data from the Central Pennsylvania Women's Health Study, which included a random-digit-dialed survey of women aged 18 to 45 years. Hierarchical multiple linear and logistic regression models were estimated to predict 3 mental health outcomes: score on a mental health measure, depressive symptoms, and diagnosed depression or anxiety. RESULTS: Mental health outcomes were associated with different factors. Farm residence was associated with higher mental health score, and the most isolated rural residence was associated with less diagnosed depression or anxiety. Elevated psychosocial stress was consistently significant across all models. A key stress modifier, self-esteem, was also consistently significant across models. Other variables associated with 2 of the outcomes were intimate partner violence exposure and affectionate social support. CONCLUSIONS: Farm residence may be protective of general mental health for women of reproductive age, and residence in isolated rural areas may decrease access to mental health screening and treatment, resulting in fewer diagnoses of depression or anxiety. PMID- 18511739 TI - Nursing home evacuation plans. AB - OBJECTIVES: I examined evacuation plans from 2134 nursing homes and analyzed national data to determine the types of nursing homes cited for deficiencies in their evacuation plans. METHODS: Evacuation plans were assessed according to criteria developed by an expert panel funded by the Office of the Inspector General. Deficiency citations came from the Online Survey, Certification, and Recording database, collected from 1997 to 2005. Four specific citations, for written emergency plans, staff training, written evacuation plans, and fire drills, were examined with multivariate logistic regression. RESULTS: Most plans had water supply provisions (96%). Only 31% specified an evacuation route. The rate of citations was relatively stable throughout the study period: each year approximately 0.6% of facilities were found to be deficient in written emergency plans, 2.1% in staff training, 1.2% in written evacuation plans, and 7.9% in fire drills. CONCLUSIONS: Some nursing homes need more specific evacuation plans. Water supply was the most and evacuation routes were the least well-addressed areas. PMID- 18511740 TI - Candidate genes and sensory functions in health and irritable bowel syndrome. AB - Adrenergic and serotonergic (ADR-SER) mechanisms alter gut (GI) function; these effects are mediated through G protein transduction. Candidate genetic variations in ADR-SER were significantly associated with somatic scores in irritable bowel syndrome (IBS) and gastric emptying but not small bowel or colonic transit. Our aim was to assess whether candidate ADR-SER genes are associated with motor and sensory GI functions in IBS and subgroups on the basis of bowel dysfunction. In 122 patients with IBS and 39 healthy controls, we assessed gastrointestinal somatic symptoms and affect by validated questionnaires. We measured: gastric volume (GV), maximum tolerated volume, rectal compliance, sensation thresholds and ratings, and genetic variations including alpha2A (C-1291G), alpha2C (Del 332 325), GNbeta3 (C825T), and 5-HTTLPR. Demographics and genotype distributions were similar in the patients with IBS subgrouped on bowel function. There were significant associations between 5-HTTLPR SS genotype and absence of IBS symptoms and between 5-HTTLPR LS/SS genotype and increased rectal compliance and increased pain ratings, particularly at 12 and 24 mmHg distensions. GNbeta3 was associated only with fasting GV; we did not detect associations between alpha2A genotype and the gastrointestinal sensory or motor functions tested. We concluded that 5 HTTLPR LS/SS genotype is associated with both increased pain sensation and increased rectal compliance though the latter effect is unlikely to contribute to increased pain sensation ratings with LS/SS genotype. The data suggest the hypotheses that the endophenotype of visceral hypersensitivity in IBS may be partly related to genetic factors, and the association of GNbeta3 with fasting GV may explain, in part, the reported association of GNbeta3 with dyspepsia. PMID- 18511741 TI - Transcriptional activation by growth hormone of HNF-6-regulated hepatic genes, a potential mechanism for improved liver repair during biliary injury in mice. AB - Growth hormone (GH) function is mediated through multiple endocrine pathways. In the liver, GH also transcriptionally activates hepatocyte nuclear factor-6 (HNF 6; OC-1), a liver-enriched transcription factor that regulates the expression of genes essential to hepatic function. We hypothesize that GH modulates hepatic function in the normal and injured liver through HNF-6 and HNF-6 target genes. CD1 mice received PBS or GH for the 1-, 7-, and 28-day course of Sham operation or bile duct ligation (BDL). Proliferation-, metabolic-, and profibrotic-specific hepatic functions were assessed with a focus on candidate HNF-6 transcriptional target genes. Confirmation of HNF-6 regulation was done by analysis of target gene expression in liver infected with recombinant adenovirus AdHNF-6 expression vectors. GH administration upregulated HNF-6 expression throughout the course of liver injury. This was associated with increased expression of HNF-6 proliferative target genes cyclin D1 and metabolic gene Cyp7A1 and downregulation of profibrogenic TGFb2R. Hepatic function improved such as enhanced hepatocyte proliferation, higher cholesterol clearance throughout the course of injury, and attenuated fibrogenic response at day 28 of BDL. GH treatment also transcriptionally increased albumin expression in an HNF-6-independent manner. This was associated with enhanced serum albumin levels. In conclusion, the GH/HNF 6 axis is a potential in vivo mechanism underlying GH diverse function in the liver to modulate the liver repair response to BDL. PMID- 18511742 TI - A synthetic prostone activates apical chloride channels in A6 epithelial cells. AB - The bicyclic fatty acid lubiprostone (formerly known as SPI-0211) activates two types of anion channels in A6 cells. Both channel types are rarely, if ever, observed in untreated cells. The first channel type was activated at low concentrations of lubiprostone (<100 nM) in >80% of cell-attached patches and had a unit conductance of approximately 3-4 pS. The second channel type required higher concentrations (>100 nM) of lubiprostone to activate, was observed in approximately 30% of patches, and had a unit conductance of 8-9 pS. The properties of the first type of channel were consistent with ClC-2 and the second with CFTR. ClC-2's unit current strongly inwardly rectified that could be best fit by models of the channel with multiple energy barrier and multiple anion binding sites in the conductance pore. The open probability and mean open time of ClC-2 was voltage dependent, decreasing dramatically as the patches were depolarized. The order of anion selectivity for ClC-2 was Cl > Br > NO(3) > I > SCN, where SCN is thiocyanate. ClC-2 was a "double-barreled" channel favoring even numbers of levels over odd numbers as if the channel protein had two conductance pathways that opened independently of one another. The channel could be, at least, partially blocked by glibenclamide. The properties of the channel in A6 cells were indistinguishable from ClC-2 channels stably transfected in HEK293 cells. CFTR in the patches had a selectivity of Cl > Br >> NO(3) congruent with SCN congruent with I. It outwardly rectified as expected for a single-site anion channel. Because of its properties, ClC-2 is uniquely suitable to promote anion secretion with little anion reabsorption. CFTR, on the other hand, could promote either reabsorption or secretion depending on the anion driving forces. PMID- 18511743 TI - Progesterone stimulates the proliferation of female and male cholangiocytes via autocrine/paracrine mechanisms. AB - During cholestatic liver diseases, cholangiocytes express neuroendocrine phenotypes and respond to a number of hormones and neuropeptides by paracrine and autocrine mechanisms. We examined whether the neuroendocrine hormone progesterone is produced by and targeted to cholangiocytes, thereby regulating biliary proliferation during cholestasis. Nuclear (PR-A and PR-B) and membrane (PRGMC1, PRGMC2, and mPRalpha) progesterone receptor expression was evaluated in liver sections and cholangiocytes from normal and bile duct ligation (BDL) rats, and NRC cells (normal rat cholangiocyte line). In vivo, normal rats were chronically treated with progesterone for 1 wk, or immediately after BDL, rats were treated with a neutralizing progesterone antibody for 1 wk. Cholangiocyte growth was measured by evaluating the number of bile ducts in liver sections. The expression of the progesterone synthesis pathway was evaluated in liver sections, cholangiocytes and NRC. Progesterone secretion was evaluated in supernatants from normal and BDL cholangiocytes and NRC. In vitro, NRC were stimulated with progesterone and cholangiocyte supernatants in the presence or absence of antiprogesterone antibody. Aminoglutethimide was used to block progesterone synthesis. Cholangiocytes and NRC express the PR-B nuclear receptor and PRGMC1, PRGMC2, and mPRalpha. In vivo, progesterone increased the number of bile ducts of normal rats, whereas antiprogesterone antibody inhibited cholangiocyte growth stimulated by BDL. Normal and BDL cholangiocytes expressed the biosynthetic pathway for and secrete progesterone. In vitro, 1) progesterone increased NRC proliferation; 2) cholangiocyte supernatants increased NRC proliferation, which was partially inhibited by preincubation with antiprogesterone; and 3) inhibition of progesterone steroidogenesis prevented NRC proliferation. In conclusion, progesterone may be an important autocrine/paracrine regulator of cholangiocyte proliferation. PMID- 18511744 TI - Regulation of direct transintestinal cholesterol excretion in mice. AB - Biliary secretion is generally considered to be an obligate step in the pathway of excess cholesterol excretion from the body. We have recently shown that an alternative route exists. Direct transintestinal cholesterol efflux (TICE) contributes significantly to cholesterol removal in mice. Our aim was to investigate whether the activity of this novel pathway can be influenced by dietary factors. In addition, we studied the role of cholesterol acceptors at the luminal side of the enterocyte. Mice were fed a Western-type diet (0.25% wt/wt cholesterol; 16% wt/wt fat), a high-fat diet (no cholesterol; 24% wt/wt fat), or high-cholesterol diet (2% wt/wt), and TICE was measured by isolated intestinal perfusion. Bile salt-phospholipid mixtures served as cholesterol acceptor. Western-type and high-fat diet increased TICE by 50 and 100%, respectively. In contrast, the high-cholesterol diet did not influence TICE. Intestinal scavenger receptor class B type 1 (Sr-B1) mRNA and protein levels correlated with the rate of TICE. Unexpectedly, although confirming a role for Sr-B1, TICE was significantly increased in Sr-B1-deficient mice. Apart from the long-term effect of diets on TICE, acute effects by luminal cholesterol acceptors were also investigated. The phospholipid content of perfusate was the most important regulator of TICE; bile salt concentration or hydrophobicity of bile salts had little effect. In conclusion, TICE can be manipulated by dietary intervention. Specific dietary modifications might provide means to stimulate TICE and, thereby, to enhance total cholesterol turnover. PMID- 18511745 TI - Heritability of longevity in captive populations of nondomesticated mammals and birds. AB - We used variance components analysis and offspring-parent regression to estimate the heritability of age at death in zoo populations of several species of mammals and birds. A meta-analysis over 14 species of mammals indicated a variance component heritability of 0.53. More conservative regression estimates of heritability for the same species averaged 0.17. Offspring-parent regressions were not significant for any of eight species of birds. Heritabilities for data simulated with frailty and age-at-death models showed that sources of variation in age at death cannot be distinguished from observed heritabilities. The CV(A) in age at death in six mammal species, based on parent-offspring regression, ranged from 0.20 to 0.54. The absence of substantial genetic variation for age at death in birds might be related to the stringency of flight, allowing for little variation in the optimization of life-history trade-offs. PMID- 18511746 TI - Adiponectin levels and genotype: a potential regulator of life span in humans. AB - Although caloric restriction in numerous models extends life, longevity in humans is suggested to be limited by the increased prevalence of obesity. Adiponectin, a fat-derived peptide, has a protective role against age-related disease, and thus is an excellent candidate gene for longevity. We studied adiponectin levels in centenarians (n = 118), their offspring (n = 228), and unrelated participants <95 (n = 78). Adiponectin levels were significantly greater in participants older than 95 years (p =.01), an effect that was independent of sex and body mass index (BMI). Adiponectin levels in the offspring were higher (following adjustment for age, sex, and BMI) compared to controls (p =.02), suggesting that inherited factors play a role in determining adiponectin levels. Over-representation of two common variants in Adiponectin gene (ADIPOQ) in male long-lived individuals combined with their independent association with elevated plasma adiponectin levels (in men and women) suggests that their presence may promote increased life span through the regulation of adiponectin production and/or secretion. PMID- 18511747 TI - A genetic-demographic approach reveals male-specific association between survival and tumor necrosis factor (A/G)-308 polymorphism. AB - The (A/G)-308 polymorphism of the tumor necrosis factor alpha gene (TNF) is associated with age-related diseases, but its influence on longevity is controversial. We genotyped for this polymorphism 747 Italian volunteers (401 women and 346 men, age 19-110 years). By applying a genetic-demographic (GD) approach we found that, in men, the survival function of allele A carriers is lower than that of noncarriers at all the ages (p =.044). After defining (by exploiting again demographic information) three age classes, we found that the frequency of men carrying the A allele decreases with age (p =.019), thus confirming the GD analysis results. The same analyses gave negative results in women. Therefore, allele A has a detrimental effect on life expectancy, and this effect is specific to men. A haplotype analysis carried out in men by screening the TNFa, TNFc, and TNFe microsatellite polymorphisms (spanning about 20 kb) confirmed the association of the TNF region with life expectancy. PMID- 18511749 TI - A percentage analysis of the telomere length in Parkinson's disease patients. AB - Telomeres are the repeated sequences at the chromosome ends which undergo shortening with cell division. The telomere shortening of the peripheral leukocytes is also facilitated by enhanced oxidative stress in various kinds of disease including ischemic heart disease, diabetes mellitus, apoplexy, and Alzheimer's disease. Telomere shortening in Parkinson's disease (PD) has not yet been reported. The pathogenesis for PD is also regarded to be associated with oxidative stress. We investigated 28 Japanese male PD patients ages 47-69. Although we could not find a statistical difference in the mean telomere length of peripheral leukocytes between the PD patients and the control participants, we found the mean telomere lengths to be shorter than 5 kb in only the PD patients and a significant PD-associated decrease in the telomeres with a length ranging from 23.1 to 9.4 kb in the patients in their 50s and 60s. These observations suggest that telomere shortening is accelerated in PD patients in comparison to the normal population. PMID- 18511748 TI - Muscle disuse: adaptation of antioxidant systems is age dependent. AB - This study investigated the age effect on antioxidant adaptation to muscle disuse. Adult and old rats were randomized into 4 groups: weight bearing (control), 3 days of hind-limb unloading (HU), 7 days of HU, and 14 days of HU. Activities of Cu-Zn superoxide dismutase (SOD), catalase, and glutathione (GSH), as well as GSH peroxidase levels were measured in the soleus. Neither disuse nor aging changed the activity of Cu-Zn SOD. The old rats had greater GSH peroxidase activity, whereas the activity of catalase had a compensatory increase with disuse, independent of age. Reduced GSH level and total glutathione (tGSH) level had age-related change with disuse. In old rats, the GSH and tGSH levels were lower with disuse, whereas the levels remained stable with disuse in adult rats. The depletion of intracellular GSH and tGSH levels of muscles from aged animals with disuse may make aged muscles more susceptible to oxidative damage. PMID- 18511750 TI - Religious involvement and healthy cognitive aging: patterns, explanations, and future directions. PMID- 18511751 TI - Church attendance mediates the association between depressive symptoms and cognitive functioning among older Mexican Americans. AB - BACKGROUND: The objective of this study was to examine how the effect of depressive symptoms on cognitive function is modified by church attendance. METHODS: We used a sample of 2759 older Mexican Americans. Cognitive function was assessed using the Mini-Mental State Examination (MMSE) at baseline, 2, 5, 7, and 11 years of follow-up. Church attendance was dichotomized as frequent attendance (e.g., going to church at least once a month) versus infrequent attendance (e.g., never or several times a year). Depressive symptoms were assessed by the Center for Epidemiologic Studies Depression Scale (CES-D; score >or=16 vs <16). General linear mixed models with time-dependent covariates were used to explore cognitive change at follow-up. RESULTS: In unadjusted models, infrequent church attendees had a greater decline in MMSE scores (drop of 0.151 points more each year, standard error [SE] = 0.02, p <.001) compared to frequent church attendees; participants having CES-D scores >or=16 also had greater declines in MMSE scores (drop of 0.132 points more each year, SE = 0.03, p <.001) compared to participants with CES-D score <16 at follow-up. In fully adjusted models, a significant Church attendance x CES-D x Time interaction (p =.001) indicated that, among participants with CES-D scores >or=16, infrequent church attendees had greater decline in MMSE scores (drop of 0.236 points more each year, SE = 0.05, p <.001) compared to frequent church attendees at follow-up. CONCLUSION: Church attendance appears to be beneficial for maintaining cognitive function of older persons. Church attendance moderates the impact of clinically relevant depressive symptoms on subsequent cognitive function. PMID- 18511753 TI - Cognitive function and oral health among community-dwelling older adults. AB - BACKGROUND: Both oral health problems and cognitive impairment are relatively common among older adults. Poorer oral health appears to contribute to a decline in quality of life and to be related to various medical conditions. Little is known about the relationship of cognitive function to oral health among community dwelling older adults. METHODS: The sample included 1984 dentate community dwelling older adults 60 years old or older from the National Health and Nutrition Examination Survey (NHANES, 1999-2002) who completed both the study cognitive measure and dental examination. Weighted descriptive and multivariate regression analyses were performed. RESULTS: Multivariate analyses showed that cognitive function was associated with oral health. Individuals with lower cognitive scores had a higher number of decayed and missing teeth and a higher proportion of periodontitis sites. The predicted number of decayed teeth increased by 0.01 with each 1-point decrease in the Digit Symbol Substitution Test score; the number of missing teeth increased by 0.02; and the percentage of sites with periodontal disease increased by 0.02. In addition, individuals' sociodemographic characteristics, health behavior, and regular dental checkups were significantly associated with oral health. CONCLUSIONS: This study suggests that community-dwelling elders with lower cognitive function scores have greater deterioration of oral health. This study provides a preliminary knowledge base for the development of early intervention strategies to address oral health problems among older adults. PMID- 18511752 TI - Adverse outcomes and correlates of change in the Short Physical Performance Battery over 36 months in the African American health project. AB - BACKGROUND: The Short Physical Performance Battery (SPPB) is a well-established measure of lower body physical functioning in older persons but has not been adequately examined in African Americans or younger persons. Moreover, factors associated with changes in SPPB over time have not been reported. METHODS: A representative sample of 998 African Americans (49-65 years old at baseline) living in St. Louis, Missouri were followed for 36 months to examine the predictive validity of SPPB in this population and identify factors associated with changes in SPPB. SPPB was calibrated to this population, ranged from 0 (worst) to 12 (best), and required imputation for about 50% of scores. Adverse outcomes of baseline SPPB included death, nursing home placement, hospitalization, physician visits, incident basic and instrumental activity of daily living disabilities, and functional limitations. Changes in SPPB over 36 months were modeled. RESULTS: Adjusted for appropriate covariates, weighted appropriately, and using propensity scores to address potential selection bias, baseline SPPB scores were associated with all adverse outcomes except physician visits, and were marginally associated with hospitalization. Declines in SPPB scores were associated with low falls efficacy (b = -1.311), perceived income adequacy (-0.121), older age (-0.073 per year), poor vision (-0.754), diabetes mellitus (-0.565), refusal to report household income (1.48), ever had Medicaid insurance (-0.610), obesity (-0.437), hospitalization in the prior year (-0.521), and kidney disease (-.956). CONCLUSIONS: The effect of baseline SPPB on adverse outcomes in this late middle-age African American population confirms reports involving older, primarily white participants. Alleviating deterioration in lower body physical functioning guided by the associated covariates may avoid or delay multiple age-associated adverse outcomes. PMID- 18511754 TI - Total and regional gray matter volume is not related to APOE*E4 status in a community sample of middle-aged individuals. AB - BACKGROUND: The APOE*E4 allele has been associated with greater gray matter atrophy and with Alzheimer's disease. The aim of this study was to investigate whether the relationship between cerebral gray matter atrophy and APOE*E4 genotype was also present in a community-dwelling, nondemented 60- to 64-year-old cohort. METHODS: Hippocampal and amygdalar volumes were manually traced and analyzed on 331 cranial T1-weighted magnetic resonance imaging (MRI) scans to detect differences associated with APOE*E4 genotype. Voxel-based morphometric (VBM) analyses were applied to detect regional gray matter volume differences. RESULTS: No total, hippocampal, or amygdalar gray matter volume difference was detected between APOE*E4 carriers and noncarriers. CONCLUSIONS: In nondemented 60 to 64-year-olds, there was no association between APOE genotype and gray matter volume using both region-of-interest analysis and VBM. PMID- 18511756 TI - Association of blood pressure and genetic background with white matter lesions in patients with mild cognitive impairment. AB - Background. White matter lesions (WMLs) may contribute to cognitive deficits in patients with mild cognitive impairment (MCI), but their pathogenesis is complex. Fluctuations of blood pressure (BP) over 24 hours and genetic predisposition to develop vascular damage have been implicated. Methods. In 63 MCI patients 65 years old or older, BP was measured both clinically and with ambulatory BP monitoring. Patients were classified in two groups: no/very mild (n = 34) and mild to severe (n = 29) WMLs, based on a visual scale on magnetic resonance (mean age 71.8 +/- 4.7 vs 74.6 +/- 5.1, and female gender 53% vs 66%, respectively). The volume of WMLs was measured by a semi-automatic method, separately for periventricular caps and rim, periventricular confluent, subcortical punctate, and subcortical confluent. Polymorphisms of cystatin C (CST3) and cholesterol 24 hydroxylase (CYP46) genes, putative risk factors for cerebrovascular disease, were determined. Results. The prevalence of cerebrovascular risk factors was similar in the two MCI groups of different WML severity, as well as clinic and ambulatory BP. In patients with mild to severe, but not in those with no/very mild WMLs, the volume of periventricular confluent WMLs increased with increasing daytime systolic BP (regression coefficient.47, 95% confidence interval [CI],.13 to.71 vs.02, 95% CI, -.32 to.36, p =.003 for the difference between slopes). The volume of other WML subtypes was not associated with ambulatory BP. Participants carrying both CST3*B and CYP46*T alleles were overrepresented in the MCI group with mild to severe WMLs (43% vs 17%, p.03). Conclusions. BP and gene putative risk factors for cerebrovascular disease are differentially associated with WMLs in two MCI groups of different WML severity. WMLs might develop for the convergence of innate with acquired factors. PMID- 18511755 TI - Glutathione peroxidase enzyme activity in aging. AB - BACKGROUND: It is hypothesized that free radical damage contributes to aging. Age related decline in activity of the antioxidant enzyme glutathione peroxidase (GPx) may contribute to increased free radicals. We hypothesized that GPx activity decreases with age in a population of older women with disability. METHODS: Whole blood GPx activity was measured in baseline stored samples from participants in the Women's Health and Aging Study I, a cohort of disabled community-dwelling older women. Linear regression was used to determine cross sectional associations between GPx activity and age, adjusting for hemoglobin, coronary disease, diabetes, selenium, and body mass index. RESULTS: Six hundred one participants had complete demographic, disease, and laboratory information. An inverse association was observed between GPx and age (regression coefficient = -2.9, p <.001), indicating that for each 1-year increase in age, GPx activity decreased by 2.9 micromol/min/L. This finding remained significant after adjustment for hemoglobin, coronary disease, diabetes, and selenium, but not after adjustment for body mass index and weight loss. CONCLUSION: This is the first study to examine the association between age and GPx activity in an older adult cohort with disability and chronic disease. These findings suggest that, after age 65, GPx activity declines with age in older women with disability. This decline does not appear to be related to diseases that have been previously reported to alter GPx activity. Longitudinal examination of GPx activity and other antioxidant enzymes in diverse populations of older adults will provide additional insight into age- and disease-related changes in these systems. PMID- 18511757 TI - Usefulness of frailty markers in the assessment of the health and functional status of older cancer patients referred for chemotherapy: a pilot study. AB - BACKGROUND: Older cancer patients seen in an oncology clinic seem to be healthier and less disabled than traditional geriatric patients. Choosing the most sensitive tools to assess their health status is a major issue. This cross sectional study explores the usefulness of frailty markers in detecting vulnerability in older cancer patients. METHODS: The study included cancer patients >or=70 years old referred to an oncology clinic for chemotherapy. Information on comorbidities, disability in instrumental activities of daily living (IADL) and activities of daily living (ADL), and seven frailty markers (nutrition, mobility, strength, energy, physical activity, mood, and cognition) was collected. Patients were classified into four hierarchical groups: 1- No frailty markers, IADL, or ADL disability; 2- Presence of frailty markers without IADL or ADL disability; 3- IADL disability without ADL disability; 4- ADL disability. RESULTS: Among the 50 patients assessed, 6 (12.0%) were classified into Group 1, 21 (42.0%) into Group 2, 15 (30.0%) into Group 3, and 8 (16.0%) into Group 4. In Group 2, 7 patients (33.3 %) had one frailty marker, and 14 (66.7%) had two or more. The most prevalent of the frailty markers were nutrition, mobility, and physical activity. CONCLUSION: The assessment of seven frailty markers allowed the detection of potential vulnerability among 42% of older cancer patients that would not have been detected through an assessment of IADL and ADL disability alone. A longitudinal study is needed to determine whether the use of frailty markers can better characterize the older cancer population and predict adverse outcomes due to cancer treatment. PMID- 18511758 TI - Balance impairment not predictive of falls in geriatric rehabilitation wards. AB - BACKGROUND: Falls are common among hospital inpatients, particularly in rehabilitation wards. Standing balance impairment is widely held to be a contributing factor to falls, is a component of several falls risk screening tools, and has motivated the development of balance retraining programs for the reduction of in-hospital falls. Little rigorous investigation of the link between standing balance impairment and in-hospital falls has been undertaken. METHODS: We identified optimal cut-off points of four commonly used balance measures (functional reach, Timed Up and Go, step test, and timed static stance) in a prospective multicenter cohort study. Admission data (n = 1373) were clustered and matched by center then randomly allocated to development and validation data sets. RESULTS: Optimal cut-off points for each test were identified from the development data set. The predictive accuracy of all four balance tests was poor when the optimal cut-off was applied to the validation data set (Youden Index scores ranged between 0.02 and 0.15). CONCLUSIONS: These findings do not support an association between admission standing balance and falls in a geriatric rehabilitation setting. This result has implications for content of falls risk screening tools and interventions to prevent falls in a geriatric rehabilitation population. PMID- 18511759 TI - Physical activity, physical function, and incident dementia in elderly men: the Honolulu-Asia Aging Study. AB - BACKGROUND: Although evidence is accumulating for a protective effect of late life physical activity on the risk of dementia, the findings are inconsistent, especially in men. We examined the association of late life physical activity and the modifying effect of physical function with future risk of dementia in a well characterized cohort of elderly men participating in the Honolulu-Asia Aging Study (HAAS). METHODS: Physical activity by self-report and performance-based physical function was assessed in 2263 men aged 71-92 years without dementia at the baseline examination of the HAAS in 1991-1993. Follow-up for incident dementia occurred at repeat examinations conducted in 1994-1996 and 1997-1999. Analyses were based on Cox proportional hazards models with adjustment for potential confounders, including age, baseline cognitive function, education, and apolipoprotein E genotype. RESULTS: There were 173 incident cases of dementia with a mean follow-up of 6.1 years. Although the incidence of dementia tended to decline with increasing physical activity and function, there was a significant interaction between the latter two factors on dementia risk (p =.022). For men with low physical function, high levels of physical activity were associated with half the risk of dementia versus men who were the least active (hazard ratio [HR], 0.50; 95% confidence interval [CI], 0.28-0.89), with a moderate level of physical activity also providing a protective effect (HR, 0.57; 95% CI, 0.32 0.99). Risk of dementia and Alzheimer's disease declined significantly with increasing physical activity. Findings persisted after age and risk factor adjustment. Similar associations were absent in men with moderate and high physical function. CONCLUSIONS: In elderly men with poor physical function, increasing general physical activity may potentially confer a protective effect or delay the onset for dementia. PMID- 18511760 TI - Altered growth hormone, cortisol, and leptin secretion in healthy elderly persons with sarcopenia and mixed body composition phenotypes. AB - BACKGROUND: Obese phenotypes and aging are independently associated with hypothalamic-pituitary-adrenocortical (HPA) axis and leptin secretion alterations. However, leptin secretion and HPA axis function in elderly persons with other body composition phenotypes is largely unknown. METHODS: Forty-five healthy elderly participants were classified normal lean (NL), sarcopenic (SS), sarcopenic-obese (SO), or obese (OO) using dual-energy x-ray absorptiometry. Growth hormone (GH), cortisol, and leptin secretion were evaluated during a free running night, and oral glucocorticoid suppression test (dexamethasone DEX). Diurnal cortisol secretion was assessed by hourly salivary samples with timed meals. Data were analyzed using cluster, deconvolution, and approximate entropy (ApEn) analyses. RESULTS: GH area, total secretion, and mean concentration during the free-running night was lower in the SO and OO groups verses the SS and NL groups (p <.02, Wilcoxon test). GH mean concentration and total secretion significantly increased in all groups during DEX (overall p <.05) except the SO group, in which ApEn increased (p =.03). Pre- and postbreakfast peak salivary cortisol (p =.004) and area under the curve (p =.03) was greatest in the SS group. Baseline leptin (11:00 pm) was significantly higher in the SO, OO, and SS groups verses the NL group (p =.01). Appendicular skeletal muscle mass was independently and negatively correlated with leptin in all groups, even after adjusting for percentage body fat (p =.001). CONCLUSIONS: In the presence of obesity, GH secretion was depressed with a blunted and disorderly response to oral glucocorticoid suppression in SO participants. Sarcopenic participants had concomitantly elevated leptin and cortisol relative to their low body fat mass. Complex or dysregulated neuroendocrine feedback systems appear to be operating in elderly persons with specific body composition phenotypes. PMID- 18511761 TI - Response to special medical sciences section on unconventional views of frailty. PMID- 18511763 TI - Predictive accuracy of falls risk screening tools. PMID- 18511764 TI - Patients' attitudes to the summary care record and HealthSpace: qualitative study. AB - OBJECTIVE: To document the views of patients and the public towards the summary care record (SCR, a centrally stored medical record drawn from the general practice record) and HealthSpace (a personal health organiser accessible through the internet from which people can view their SCR), with a particular focus on those with low health literacy, potentially stigmatising conditions, or difficulties accessing health care. DESIGN: 103 semistructured individual interviews and seven focus groups. SETTING: Three early adopter primary care trusts in England where the SCR and HealthSpace are being piloted. All were in areas of relative socioeconomic deprivation. PARTICIPANTS: Individual participants were recruited from general practice surgeries, walk-in centres, out of hours centres, and accident and emergency departments. Participants in focus groups were recruited through voluntary sector organisations; they comprised advocates of vulnerable groups and advocates of people who speak limited English; people with HIV; users of mental health services; young adults; elderly people; and participants of a drug rehabilitation programme. METHODS: Participants were asked if they had received information about the SCR and HealthSpace and about their views on shared electronic records in different circumstances. RESULTS: Most people were not aware of the SCR or HealthSpace and did not recall receiving information about it. They saw both benefits and drawbacks to having an SCR and described a process of weighing the former against the latter when making their personal choice. Key factors influencing this choice included the nature of any illness (especially whether it was likely to lead to emergency care needs); past and present experience of healthcare and government surveillance; the person's level of engagement and health literacy; and their trust and confidence in the primary healthcare team and the wider NHS. Overall, people with stigmatising illness were more positive about the SCR than people who claimed to speak for "vulnerable groups." Misconceptions about the SCR were common, especially confusion about what data it contained and who would have access to it. Most people were not interested in recording their medical data or accessing their SCR via HealthSpace, but some saw the potential for this new technology to support self management and lay care for those with chronic illness. CONCLUSION: Despite an extensive information programme in early adopter sites, the public remains unclear about current policy on shared electronic records, though most people view these as a positive development. The "implied consent" model for creating and accessing a person's SCR should be revisited, perhaps in favour of "consent to view" at the point of access. PMID- 18511765 TI - Adherence to Mediterranean diet and risk of developing diabetes: prospective cohort study. AB - OBJECTIVE: To assess the relation between adherence to a Mediterranean diet and the incidence of diabetes among initially healthy participants. DESIGN: Prospective cohort study with estimates of relative risk adjusted for sex, age, years of university education, total energy intake, body mass index, physical activity, sedentary habits, smoking, family history of diabetes, and personal history of hypertension. SETTING: Spanish university department. PARTICIPANTS: 13 380 Spanish university graduates without diabetes at baseline followed up for a median of 4.4 years. MAIN OUTCOME MEASURES: Dietary habits assessed at baseline with a validated 136 item food frequency questionnaire and scored on a nine point index. New cases of diabetes confirmed through medical reports and an additional detailed questionnaire posted to those who self reported a new diagnosis of diabetes by a doctor during follow-up. Confirmed cases of type 2 diabetes. RESULTS: Participants who adhered closely to a Mediterranean diet had a lower risk of diabetes. The incidence rate ratios adjusted for sex and age were 0.41 (95% confidence interval 0.19 to 0.87) for those with moderate adherence (score 3 6) and 0.17 (0.04 to 0.75) for those with the highest adherence (score 7-9) compared with those with low adherence (score <3). In the fully adjusted analyses the results were similar. A two point increase in the score was associated with a 35% relative reduction in the risk of diabetes (incidence rate ratio 0.65, 0.44 to 0.95), with a significant inverse linear trend (P=0.04) in the multivariate analysis. CONCLUSION: Adherence to a Mediterranean diet is associated with a reduced risk of diabetes. PMID- 18511767 TI - Reforming NHS dentistry. PMID- 18511768 TI - Top-up fees: Top-up fairly according to means. PMID- 18511769 TI - Top-up fees: Not allowing top-up fees is unethical. PMID- 18511770 TI - All cause mortality: Meaningless without time frame. PMID- 18511771 TI - Blunt abdominal trauma: Trauma services must improve. PMID- 18511772 TI - The secret lives of doctors: Inspired by disease and death. PMID- 18511773 TI - Writing on the wall for UNAIDS: Let's use our tools creatively. PMID- 18511774 TI - Writing on the wall for UNAIDS: Consequences of mythology. PMID- 18511775 TI - Writing on the wall for UNAIDS: UNAIDS replies. PMID- 18511776 TI - Unnecessary emergency visits: Middle East has an answer. PMID- 18511778 TI - Google launches free electronic health records service for patients. PMID- 18511779 TI - Italian police arrest drug agency officials over alleged falsification of data. PMID- 18511780 TI - Merck to pay $58m in settlement over rofecoxib advertising. PMID- 18511781 TI - Health services in China face enormous challenge after earthquake. PMID- 18511783 TI - Prison mental health services are underfunded, report says. PMID- 18511784 TI - Finding a solution to hospital associated infections. PMID- 18511786 TI - Poor countries urgently need to scale up training of healthcare workers, task force warns. PMID- 18511787 TI - FDA begins active monitoring of drug safety. PMID- 18511790 TI - Patchy blood testing in central Asia raises risk of HIV infection. PMID- 18511791 TI - Number of alcohol related admissions in England has doubled in 12 years. PMID- 18511792 TI - Report names hospitals with highest proportions of deaths related to MRSA and C difficile. PMID- 18511793 TI - East Africa faces starvation as rising food prices worsen effect of war and drought. PMID- 18511794 TI - Leaping to conclusions. PMID- 18511795 TI - Let's not widen the gulf in the health care of children. PMID- 18511796 TI - Founding principles. PMID- 18511797 TI - Is there an epidemic of admissions for surgical treatment of dental abscesses in the UK? PMID- 18511798 TI - Management of retinal detachment: a guide for non-ophthalmologists. PMID- 18511799 TI - Audit: how to do it in practice. PMID- 18511800 TI - Risk assessment and lipid modification for primary and secondary prevention of cardiovascular disease: summary of NICE guidance. PMID- 18511801 TI - Commentary: Controversies in NICE guidance on lipid modification for the prevention of cardiovascular disease. PMID- 18511802 TI - The case for resurrecting the long case. PMID- 18511806 TI - CC chemokine receptor 5 gene promoter activation by the cyclic AMP response element binding transcription factor. AB - The chemokine receptor CCR5 is implicated in the pathogenesis of various inflammatory diseases, such as multiple sclerosis (MS), atherosclerosis, transplant rejection, and autoimmunity. In previous studies, we have shown that MS lesions are characterized by enhanced expression of transcription factors associated with stress responses, ie, IRF-1, NF-kappaB, and CREB-1, which modulate expression of both classes of major histocompatibility complex (MHC) molecules. The expression of MHC-I and MHC-II molecules greatly overlaps with the expression of CCR5 in MS lesions. Therefore, we investigated whether these factors are also involved in the transcriptional regulation of CCR5. Using in vitro assays, we determined that neither IRF-1 nor NF-kappaB is involved in the activation of the CCR5 promoter. This is corroborated by the finding that these factors are not involved in the induction of endogenous CCR5 transcription in various cell types. In contrast, we show that CCR5 expression is regulated by the cAMP/CREB pathway and that interference in this pathway affects endogenous CCR5 transcription. From this, we conclude that the cAMP/CREB pathway is involved in the regulation of CCR5 transcription and that, given the ubiquitous nature of CREB-1 protein expression, additional regulatory mechanisms must contribute to cell type-specific expression of CCR5. PMID- 18511807 TI - Central role of PI3K in transcriptional activation of hTERT in HTLV-I-infected cells. AB - The persistence of human T-cell leukemia/lymphoma virus-I (HTLV-I)-infected cells is dependent upon clonal expansion and up-regulation of telomerase (hTERT). We have previously found that in interleukin (IL)-2-independent transformed HTLV-I cells, Tax strongly activates the hTERT promoter through nuclear factor-kappaB (NF-kappaB)-mediated Sp1 and c-Myc activation. In IL-2-dependent cells and adult T-cell leukemia/lymphoma (ATLL) patient samples, however, Tax expression is very low to undetectable, yet these cells retain strong telomerase activity. This suggests the existence of compensatory mechanisms in IL-2-dependent cells and ATLL patients. In this study, we demonstrate that telomerase activity is significantly decreased upon IL-2 withdrawal in immortalized HTLV-I cell lines. Inhibition of PI3K or AKT signaling pathways reduced telomerase activity in HTLV I cells. We found that IL-2/IL-2R signaling was associated with a PI3K dependent/AKT-independent transcriptional up-regulation of the endogenous hTERT promoter. We found that activation of the PI3K pathway mediated cytoplasmic retention of the Wilms tumor (WTI) protein, which strongly suppressed the hTERT promoter. The importance of this regulatory pathway for telomerase expression is underscored by findings that the PI3K pathway is commonly found activated in cancer cells. PMID- 18511808 TI - t(8;21)(q22;q22) Fusion proteins preferentially bind to duplicated AML1/RUNX1 DNA binding sequences to differentially regulate gene expression. AB - Chromosome abnormalities are frequently associated with cancer development. The 8;21(q22;q22) chromosomal translocation is one of the most common chromosome abnormalities identified in leukemia. It generates fusion proteins between AML1 and ETO. Since AML1 is a well-defined DNA-binding protein, AML1-ETO fusion proteins have been recognized as DNA-binding proteins interacting with the same consensus DNA-binding site as AML1. The alteration of AML1 target gene expression due to the presence of AML1-ETO is related to the development of leukemia. Here, using a 25-bp random double-stranded oligonucleotide library and a polymerase chain reaction (PCR)-based DNA-binding site screen, we show that compared with native AML1, AML1-ETO fusion proteins preferentially bind to DNA sequences with duplicated AML1 consensus sites. This finding is further confirmed by both in vitro and in vivo DNA-protein interaction assays. These results suggest that AML1 ETO fusion proteins have a selective preference for certain AML1 target genes that contain multimerized AML1 consensus sites in their regulatory elements. Such selected regulation provides an important molecular mechanism for the dysregulation of gene expression during cancer development. PMID- 18511809 TI - The cytoplasmic tail of CD45 is released from activated phagocytes and can act as an inhibitory messenger for T cells. AB - CD45 is the prototypic transmembrane protein tyrosine phosphatase (PTP), which is expressed on all nucleated hematopoietic cells and plays a central role in the integration of environmental signals into immune cell responses. Here we report an alternative function for the intracellular domain of CD45. We dis-covered that CD45 is sequentially cleaved by serine/metalloproteinases and gamma-secretases during activation of human monocytes and granulocytes by fungal stimuli or phorbol 12-myristate 13-acetate but not by other microbial stimuli. Proteolytic processing of CD45 occurred upon activation of monocytes or granulocytes but not of T cells, B cells, or dendritic cells and resulted in a 95-kDa fragment of the cytoplasmic tail of CD45 (ct-CD45). ct-CD45 was released from monocytes and granulocytes upon activation-induced cell death. Binding studies with ct-CD45 revealed a counter-receptor on preactivated T cells. Moreover, T-cell proliferation induced by dendritic cells or CD3 antibodies was inhibited in the presence of ct-CD45. Taken together, the results of our study demonstrate that fragments of the intracellular domain of CD45 from human phagocytes can function as intercellular regulators of T-cell activation. PMID- 18511811 TI - A survey of hospital quality improvement activities. AB - Five years after the Institute of Medicine (IOM) called for a redesigned U.S. health care system, relatively little was known about the extent to which hospitals had undertaken quality improvement (QI) efforts to address deficiencies in patient care. To examine the state of hospital QI activities in 2006, the authors designed and conducted a survey of short-term, general hospitals with 25 or more beds. In a sample of 470 hospitals, they found that many were actively engaged in improvement efforts but that these activities varied in method and impact. Hospitals with high levels of perceived quality, as reflected in assessments by their quality managers, were more likely to have embraced QI as a strategic priority, employed quality practices and processes consistent with IOM aims, fostered staff training and involvement in QI methods, engaged in an array of QI activities and clinical QI strategies, and maintained staffing levels favoring fewer patients per nurse. PMID- 18511810 TI - A selective sphingosine kinase 1 inhibitor integrates multiple molecular therapeutic targets in human leukemia. AB - The potent bioactive sphingolipid mediator, sphingosine-1-phosphate (S1P), is produced by 2 sphingosine kinase isoenzymes, SphK1 and SphK2. Expression of SphK1 is up-regulated in cancers, including leukemia, and associated with cancer progression. A screen of sphingosine analogs identified (2R,3S,4E)-N-methyl-5-(4' pentylphenyl)-2-aminopent-4-ene-1,3-diol, designated SK1-I (BML-258), as a potent, water-soluble, isoenzyme-specific inhibitor of SphK1. In contrast to pan SphK inhibitors, SK1-I did not inhibit SphK2, PKC, or numerous other protein kinases. SK1-I decreased growth and survival of human leukemia U937 and Jurkat cells, and enhanced apoptosis and cleavage of Bcl-2. Lethality of SK1-I was reversed by caspase inhibitors and by expression of Bcl-2. SK1-I not only decreased S1P levels but concomitantly increased levels of its proapoptotic precursor ceramide. Conversely, S1P protected against SK1-I-induced apoptosis. SK1-I also induced multiple perturbations in activation of signaling and survival related proteins, including diminished phosphorylation of ERK1/2 and Akt. Expression of constitutively active Akt protected against SK1-I-induced apoptosis. Notably, SK1-I potently induced apoptosis in leukemic blasts isolated from patients with acute myelogenous leukemia but was relatively sparing of normal peripheral blood mononuclear leukocytes. Moreover, SK1-I markedly reduced growth of AML xenograft tumors. Our results suggest that specific inhibitors of SphK1 warrant attention as potential additions to the therapeutic armamentarium in leukemia. PMID- 18511812 TI - Conceptualization and measurement of organizational readiness for change: a review of the literature in health services research and other fields. AB - Health care practitioners and change experts contend that organizational readiness for change is a critical precursor to successful change implementation. This article assesses how organizational readiness for change has been defined and measured in health services research and other fields. Analysis of 106 peer reviewed articles reveals conceptual ambiguities and disagreements in current thinking and writing about organizational readiness for change. Inspection of 43 instruments for measuring organizational readiness for change reveals limited evidence of reliability or validity for most publicly available measures. Several conceptual and methodological issues that need to be addressed to generate knowledge useful for practice are identified and discussed. PMID- 18511813 TI - Molecular imaging: the future of modern medicine. PMID- 18511814 TI - Session 1: strategies to engage pharma. PMID- 18511815 TI - How the biopharmaceutical industry uses molecular imaging. PMID- 18511816 TI - Imaging in drug discovery, preclinical, and early clinical development. PMID- 18511817 TI - Imaging biomarkers, an industry perspective. PMID- 18511818 TI - Strategies to engage industry. PMID- 18511819 TI - Imaging biomarkers and surrogates: the evolving regulatory lexicon. PMID- 18511820 TI - Collaborations in the development and validation of imaging biomarkers. PMID- 18511821 TI - Session 2: strategies for fast-track technological development and regulatory issues. PMID- 18511822 TI - NCI Cancer Imaging Program update. PMID- 18511823 TI - Working together to enhance the efficiency of medical product development. PMID- 18511824 TI - The exploratory IND. PMID- 18511825 TI - Use of eINDs for evaluation of multiple related PET amyloid plaque imaging agents. PMID- 18511826 TI - Health economics in technology development: is it worth it? PMID- 18511827 TI - Session 3: strategies to engage referring physicians and increase utilization of clinical molecular imaging. PMID- 18511828 TI - A New Paradigm to Increase Utilization of PET/CT. PMID- 18511829 TI - Engaging and nurturing referring physician relationships. PMID- 18511830 TI - Steps in moving molecular imaging to the clinic. PMID- 18511831 TI - Cardiovascular molecular imaging: promoting utilization and outreach. PMID- 18511832 TI - Utilizing technological advances to grow molecular imaging clinical services: industry perspective. PMID- 18511833 TI - Individualizing cancer therapies using "anatomolecular" imaging. PMID- 18511834 TI - Session 4: strategies for getting the word out. PMID- 18511835 TI - Challenges in realizing the potential of clinical molecular imaging. PMID- 18511836 TI - Strategies in education and outreach. PMID- 18511837 TI - Making the case for molecular imaging. PMID- 18511838 TI - Professionalism and MOC. PMID- 18511839 TI - MI activities at SNM annual meeting. PMID- 18511840 TI - A look back at a year of extraordinary success and forward to a rewarding future. PMID- 18511841 TI - Reminder: hazmat personnel need to be properly trained and documented. PMID- 18511842 TI - PET imaging of cancer immunotherapy. AB - Immune system activation can be elicited in viral infections, active immunization, or cancer immunotherapy, leading to the final common phenotype of increased glycolytic use by immune cells and subsequent detection by 18F-FDG PET. Because 18F-FDG is also used in baseline staging PET/CT scans and in tumor response assessment, physicians are faced with a unique challenge when evaluating tumor response in patients receiving cancer immunotherapy. The burgeoning field of cancer immunotherapy and the paucity of PET probes that can reliably differentiate activated immune cells from metabolically active cancer cells underscore the pressing need to identify and develop additional molecular imaging strategies. In an effort to address this concern, investigators have taken several molecular imaging approaches for cancer immunotherapy. Direct ex vivo labeling of T lymphocytes with radioactive probes before reinfusion represents the earliest attempts but has proven to be clinically limited because of significant PET probe dilution from proliferation of activated immune cells. Another approach is the indirect in vivo labeling of immune cells via PET reporter gene expression and involves the ex vivo genetic engineering of T lymphocytes with a reporter gene, reinfusion into the host, and the subsequent use of a PET probe specific for the reporter gene. The most recent approach involves the direct in vivo labeling of immune cells by targeting endogenous immune cell biochemical pathways that are differentially expressed during activation. In conclusion, these novel PET-based imaging approaches have demonstrated promise toward the goal of in vivo, noninvasive immune monitoring strategies for evaluating cancer immunotherapy. PMID- 18511843 TI - Further explorations of cellular uptake of radioactivity. PMID- 18511844 TI - Impact of image-space resolution modeling for studies with the high-resolution research tomograph. AB - Brain PET in small structures is challenged by low resolution inducing bias in the activity measurements. Improved spatial resolution may be obtained by using dedicated tomographs and more comprehensive modeling of the acquisition system during reconstruction. In this study, we assess the impact of resolution modeling (RM) during reconstruction on image quality and on the estimates of biologic parameters in a clinical study performed on a high-resolution research tomograph. METHODS: An accelerated list-mode ordinary Poisson ordered-subset expectation maximization (OP-OSEM) algorithm, including sinogram-based corrections and an experimental stationary model of resolution, has been designed. Experimental phantom studies are used to assess contrast and noise characteristics of the reconstructed images. The binding potential of a selective tracer of the dopamine transporter is also assessed in anatomic volumes of interest in a 5-patient study. RESULTS: In the phantom experiment, a slower convergence and a higher contrast recovery are observed for RM-OP-OSEM than for OP-OSEM for the same level of statistical noise. RM-OP-OSEM yields contrast recovery levels that could not be reached without RM as well as better visual recovery of the smallest spheres and better delineation of the structures in the reconstructed images. Statistical noise has lower variance at the voxel level with RM than without at matched resolution. In a uniform activity region, RM induces higher positive and lower negative correlations with neighboring voxels, leading to lower spatial variance. Clinical images reconstructed with RM demonstrate better delineation of cortical and subcortical structures in both time-averaged and parametric images. The binding potential in the striatum is also increased, a result similar to the one observed in the phantom study. CONCLUSION: In high-resolution PET, RM during reconstruction improves quantitative accuracy by reducing the partial-volume effects. PMID- 18511845 TI - TGFbeta1, TNFalpha, and insulin signaling crosstalk in regulation of the rat cholesterol 7alpha-hydroxylase gene expression. AB - The TGFbeta1/Smad pathway plays a critical role in cholestasis and liver fibrosis. Previous studies show that TGFbeta1, TNFalpha, and insulin inhibit cholesterol 7alpha-hydroxylase (CYP7A1) gene transcription and bile acid synthesis in human hepatocytes. In this study, we investigated insulin, TGFbeta1, and TNFalpha regulation of rat Cyp7a1 gene transcription. In contrast to inhibition of human CYP7A1 gene transcription, TGFbeta1 stimulates rat Cyp7a1 reporter activity. Smad3, FoxO1, and HNF4alpha synergistically stimulated rat Cyp7a1 gene transcription. Mutations of the Smad3, FoxO1, or HNF4alpha binding site attenuated the rat Cyp7a1 promoter activity. Furthermore, TNFalpha and cJun attenuated TGFbeta1 stimulation of rat Cyp7a1. Insulin or adenovirus-mediated expression of constitutively active AKT1 inhibited FoxO1 and Smad3 synergy. In streptozotocin-induced diabetic rats, Cyp7a1 mRNA expression levels were induced and insulin attenuated CYP7A1 mRNA levels. Chromatin immunoprecipitation assay showed that FoxO1 binding to Cyp7a1 chromatin was increased in diabetic rat livers and insulin reduced FoxO1 binding. These results suggest a mechanistic basis for induction of Cyp7a1 activity and bile acid synthesis in cholestatic rats and in diabetic rats. The crosstalk of insulin, TGFbeta and TNFalpha signaling pathways may regulate bile acid synthesis and lipid homeostasis in diabetes, fatty liver disease, and liver fibrosis. PMID- 18511846 TI - Receptor for advanced glycation end products (RAGE) deficiency attenuates the development of atherosclerosis in diabetes. AB - OBJECTIVE: Activation of the receptor for advanced glycation end products (RAGE) in diabetic vasculature is considered to be a key mediator of atherogenesis. This study examines the effects of deletion of RAGE on the development of atherosclerosis in the diabetic apoE(-/-) model of accelerated atherosclerosis. RESEARCH DESIGN AND METHODS: ApoE(-/-) and RAGE(-/-)/apoE(-/-) double knockout mice were rendered diabetic with streptozotocin and followed for 20 weeks, at which time plaque accumulation was assessed by en face analysis. RESULTS: Although diabetic apoE(-/-) mice showed increased plaque accumulation (14.9 +/- 1.7%), diabetic RAGE(-/-)/apoE(-/-) mice had significantly reduced atherosclerotic plaque area (4.9 +/- 0.4%) to levels not significantly different from control apoE(-/-) mice (4.3 +/- 0.4%). These beneficial effects on the vasculature were associated with attenuation of leukocyte recruitment; decreased expression of proinflammatory mediators, including the nuclear factor-kappaB subunit p65, VCAM-1, and MCP-1; and reduced oxidative stress, as reflected by staining for nitrotyrosine and reduced expression of various NADPH oxidase subunits, gp91phox, p47phox, and rac-1. Both RAGE and RAGE ligands, including S100A8/A9, high mobility group box 1 (HMGB1), and the advanced glycation end product (AGE) carboxymethyllysine were increased in plaques from diabetic apoE(-/ ) mice. Furthermore, the accumulation of AGEs and other ligands to RAGE was reduced in diabetic RAGE(-/-)/apoE(-/-) mice. CONCLUSIONS: This study provides evidence for RAGE playing a central role in the development of accelerated atherosclerosis associated with diabetes. These findings emphasize the potential utility of strategies targeting RAGE activation in the prevention and treatment of diabetic macrovascular complications. PMID- 18511847 TI - Differential roles of cardiomyocyte and macrophage peroxisome proliferator activated receptor gamma in cardiac fibrosis. AB - OBJECTIVE: Cardiac fibrosis is an important component of diabetic cardiomyopathy. Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands repress proinflammatory gene expression, including that of osteopontin, a known contributor to the development of myocardial fibrosis. We thus investigated the hypothesis that PPARgamma ligands could attenuate cardiac fibrosis. RESEARCH DESIGN AND METHODS: Wild-type cardiomyocyte- and macrophage-specific PPARgamma(-/ ) mice were infused with angiotensin II (AngII) to promote cardiac fibrosis and treated with the PPARgamma ligand pioglitazone to determine the roles of cardiomyocyte and macrophage PPARgamma in cardiac fibrosis. RESULTS: Cardiomyocyte-specific PPARgamma(-/-) mice (cPPARgamma(-/-)) developed spontaneous cardiac hypertrophy with increased ventricular osteopontin expression and macrophage content, which were exacerbated by AngII infusion. Pioglitazone attenuated AngII-induced fibrosis, macrophage accumulation, and osteopontin expression in both wild-type and cPPARgamma(-/-) mice but induced hypertrophy in a PPARgamma-dependent manner. We pursued two mechanisms to explain the antifibrotic cardiomyocyte-PPARgamma-independent effects of pioglitazone: increased adiponectin expression and attenuation of proinflammatory macrophage activity. Adenovirus-expressed adiponectin had no effect on cardiac fibrosis and the PPARgamma ligand pioglitazone did not attenuate AngII-induced cardiac fibrosis, osteopontin expression, or macrophage accumulation in monocyte-specific PPARgamma(-/-) mice. CONCLUSIONS: We arrived at the following conclusions: 1) both cardiomyocyte-specific PPARgamma deficiency and activation promote cardiac hypertrophy, 2) both cardiomyocyte and monocyte PPARgamma regulate cardiac macrophage infiltration, 3) inflammation is a key mediator of AngII-induced cardiac fibrosis, 4) macrophage PPARgamma activation prevents myocardial macrophage accumulation, and 5) PPARgamma ligands attenuate AngII-induced cardiac fibrosis by inhibiting myocardial macrophage infiltration. These observations have important implications for potential interventions to prevent cardiac fibrosis. PMID- 18511848 TI - Hypothalamic protein kinase C regulates glucose production. AB - OBJECTIVE: A selective rise in hypothalamic lipid metabolism and the subsequent activation of SUR1/Kir6.2 ATP-sensitive K(+) (K(ATP)) channels inhibit hepatic glucose production. The mechanisms that link the ability of hypothalamic lipid metabolism to the activation of K(ATP) channels remain unknown. RESEARCH DESIGN AND METHODS: To examine whether hypothalamic protein kinase C (PKC) mediates the ability of central nervous system lipids to activate K(ATP) channels and regulate glucose production in normal rodents, we first activated hypothalamic PKC in the absence or presence of K(ATP) channel inhibition. We then inhibited hypothalamic PKC in the presence of lipids. Tracer-dilution methodology in combination with the pancreatic clamp technique was used to assess the effect of hypothalamic administrations on glucose metabolism in vivo. RESULTS: We first reported that direct activation of hypothalamic PKC via direct hypothalamic delivery of PKC activator 1-oleoyl-2-acetyl-sn-glycerol (OAG) suppressed glucose production. Coadministration of hypothalamic PKC-delta inhibitor rottlerin with OAG prevented the ability of OAG to activate PKC-delta and lower glucose production. Furthermore, hypothalamic dominant-negative Kir6.2 expression or the delivery of the K(ATP) channel blocker glibenclamide abolished the glucose production lowering effects of OAG. Finally, inhibition of hypothalamic PKC eliminated the ability of lipids to lower glucose production. CONCLUSIONS: These studies indicate that hypothalamic PKC activation is sufficient and necessary for lowering glucose production. PMID- 18511849 TI - Smooth muscle expression of lipoma preferred partner is mediated by an alternative intronic promoter that is regulated by serum response factor/myocardin. AB - Lipoma preferred partner (LPP) was recently recognized as a smooth muscle marker that plays a role in smooth muscle cell migration. In this report, we focus on the transcriptional regulation of the LPP gene. In particular, we investigate whether LPP is directly regulated by serum response factor (SRF). We show that the LPP gene contains 3 evolutionarily conserved CArG boxes and that 1 of these is part of an alternative promoter in intron 2. Quantitative RT-PCR shows that this alternative promoter directs transcription specifically to smooth muscle containing tissues in vivo. By using chromatin immunoprecipitation, we demonstrate that 2 of the CArG boxes, including the promoter-associated CArG box, bind to endogenous SRF in cultured aortic smooth muscle cells. Electrophoretic mobility-shift assays show that the conserved CArG boxes bind SRF in vitro. In reporter experiments, we show that the alternative promoter has transcriptional capacity that is dependent on SRF/myocardin and that the promoter associated CArG box is required for that activity. Finally, we show by quantitative RT-PCR that the alternative promoter is strongly downregulated in SRF-deficient embryonic stem cells and in smooth muscle tissues derived from conditional SRF knockout mice. Collectively, our data demonstrate that expression of LPP in smooth muscle is mediated by an alternative promoter that is regulated by SRF/myocardin. PMID- 18511850 TI - Angiogenic function of prostacyclin biosynthesis in human endothelial progenitor cells. AB - The role of prostaglandin production in the control of regenerative function of endothelial progenitor cells (EPCs) has not been studied. We hypothesized that activation of cyclooxygenase (COX) enzymatic activity and the subsequent production of prostacyclin (PGI(2)) is an important mechanism responsible for the regenerative function of EPCs. In the present study, we detected high levels of COX-1 protein expression and PGI(2) biosynthesis in human EPCs outgrown from blood mononuclear cells. Expression of COX-2 protein was almost undetectable under basal conditions but significantly elevated after treatment with tumor necrosis factor-alpha. Condition medium derived from EPCs hyperpolarized human coronary artery smooth muscle cells, similar to the effect of the PGI(2) analog iloprost. The proliferation and in vitro tube formation by EPCs were inhibited by the COX inhibitor indomethacin or by genetic inactivation of COX-1 or PGI(2) synthase with small interfering (si)RNA. Impaired tube formation and cell proliferation induced by inactivation of COX-1 were rescued by the treatment with iloprost or the selective peroxisome proliferator-activated receptor (PPAR)delta agonist GW501516 but not by the selective PGI(2) receptor agonist cicaprost. Downregulation of PPARdelta by siRNA also reduced angiogenic capacity of EPCs. Iloprost failed to reverse PPARdelta siRNA-induced impairment of angiogenesis. Furthermore, transfection of PGI(2) synthase siRNA, COX-1 siRNA, or PPARdelta siRNA into EPCs decreased the capillary formation in vivo after transplantation of human EPCs into the nude mice. These results suggest that activation of COX 1/PGI(2)/PPARdelta pathway is an important mechanism underlying proangiogenic function of EPCs. PMID- 18511851 TI - Intercellular adhesion molecule-1-dependent neutrophil adhesion to endothelial cells induces caveolae-mediated pulmonary vascular hyperpermeability. AB - We investigated the role of caveolae in the mechanism of increased pulmonary vascular permeability and edema formation induced by the activation of polymorphonuclear neutrophils (PMNs). We observed that the increase in lung vascular permeability induced by the activation of PMNs required caveolin-1, the caveolae scaffold protein. The permeability increase induced by PMN activation was blocked in caveolin-1 knockout mice and by suppressing caveolin-1 expression in rats. The response was also dependent on Src phosphorylation of caveolin-1 known to activate caveolae-mediated endocytosis in endothelial cells. To address the role of PMN interaction with endothelial cells, we used an intercellular adhesion molecule (ICAM)-1 blocking monoclonal antibody. Preventing the ICAM-1 mediated PMN binding to endothelial cells abrogated Src phosphorylation of caveolin-1, as well as the increase in endothelial permeability. Direct ICAM-1 activation by crosslinking recapitulated these responses, suggesting that ICAM-1 activates caveolin-1 signaling responsible for caveolae-mediated endothelial hyperpermeability. Our results provide support for the novel concept that a large component of pulmonary vascular hyperpermeability induced by activation of PMNs adherent to the vessel wall is dependent on signaling via caveolin-1 and increased caveolae-mediated transcytosis. Thus, it is important to consider the role of the transendothelial vesicular permeability pathway that contributes to edema formation in developing therapeutic interventions against PMN-mediated inflammatory diseases such as acute lung injury. PMID- 18511852 TI - From Darwin and Metchnikoff to Burnet and beyond. AB - Phagocytosis in unicellular animals represents the most ancient and ubiquitous form of defense against foreign material. Unicellular invertebrates can phagocytose for food and defense. Multicellular invertebrates and vertebrates possess phagocytic cells and have evolved more complex functions attributed to immunodefense cells that specialized into cellular and humoral immune responses. Thus all animals possess: innate, natural, nonspecific (no memory) nonanticipatory, nonclonal, germline (hard wired) host defense functions. In addition, all vertebrates possess: adaptive, induced, specific (memory), anticipatory, clonal, somatic (flexible) immune responses. A similar situation exists with respect to components of the signaling system, immunity and development. With multicellularity, clearly numerous immune response characteristics are not possible in unicellular forms or even those that straddle the divide between unicellularity and multicellularity, beginning with colonial/social protozoans. Still, it is instructive to elucidate a hierarchy of animals based upon immunologic characteristics and how they parallel other physiological traits. Evidence is presented that the most primitive of invertebrates prior to the evolution of multicellular organisms possess varying degrees of complexity at the molecular level of those hallmarks that now characterize the immune system. PMID- 18511853 TI - General introduction to innate immunity: Dr. Jekyl/Mr. Hyde quality of the innate immune system. AB - The innate immunity plays a critical role in host protection against pathogens and transformed cells. It relies amongst others on pattern recognition receptors such as Toll-like receptors, C-type lectin receptors, and nucleotide-binding oligomerization domain proteins to alert and activate defense pathways including the activation of the complement system. Innate immunity represents a trait common to plants and animals, and besides the humoral factors different cell types e.g. subspecies of dendritic cells (plasmacytoid dendritic cells), phagocytic cells, mast cells, glia cells, Kupffer cells, neutrophils and natural killer cells are involved to orchestrate the anti-infectious and antitumor response. Studies in plants, in fruit flies and in mammals reveal that the defensive strategies of invertebrates and vertebrates are highly conserved at the molecular level, which raises the exciting prospects of an increased understanding of innate immunity in a healthy or diseased organism. However, the molecular machinery, e.g. cytokines and chemokines, which triggers, amplifies, and sustains the different phases of the innate immune response could also promote a substantial imbalance between danger and inflammatory response when an infectious challenge is either chronic or not properly declining. PMID- 18511854 TI - The innate immune system of mammals and insects. AB - Infectious agents threaten any organism. Therefore, mammals and insects have evolved a complex network of cells and humoral factors termed immune system able to control and eliminate pathogens. Immunity varies between different groups of animals but always contains an innate immune system that can act fast and often effectively against a wide range of distinct pathogens (i.e. viruses, bacteria, fungi, and eukaryotic parasites). In mammals and insects, the communication between and regulation of immune cells is carried out by cytokines which orchestrate the defense against the invaders. The major challenge to recognize and to fight pathogens is the same for any host. In insects and mammals, the pathogens are recognized as non-self by recognition of pathogen-associated molecular patterns. In addition, similar pathogen recognition receptors and signaling pathways activate the immune response in insects and mammals. The pathogens have to be opsonized and/or ingested and controlled/eliminated by antimicrobial peptides or small effector molecules (reactive oxygen and nitrogen intermediates). Interestingly, even invertebrates have evolved certain forms of adaptive immunity, i.e. specific immune priming, and in some invertebrates alternative splicing of pathogen recognition receptors allows for a more specific recognition of a wide variety of pathogens. This enhanced specificity of pattern recognition conveys a special form of memory to their invertebrate hosts. In this chapter, we also consider gut immunity of insects and compare it with the response in mammals. PMID- 18511855 TI - Pattern recognition receptors and their role in innate immunity: focus on microbial protein ligands. AB - Antigen-presenting cells, such as macrophages and dendritic cells, represent a central and important part of the immune defence against invading microorganisms, as they participate in initial capture and processing of microbial antigens (innate immunity) and then activation of specific T and B cell effector mechanisms (acquired immunity). Recognition of microbial molecules by antigen presenting cells occurs through so called pattern recognition receptors (PRRs), which recognize conserved structures, or pathogen-associated molecular patterns, in pathogenic microbes. The Toll-like receptors are the most extensively studied of these receptors, but accumulating evidence shows that other PRRs, such as scavenger receptors, C-type lectin receptors and NOD-like receptors, also play important roles in the innate immune defence. Here, we summarize current knowledge of the role of various PRRs in the defence against pathogenic microorganisms and we report recent advances in studies of different receptor ligand interactions. In particular, we focus on the importance of microbial proteins as ligands for PRRs. PMID- 18511856 TI - Antimicrobial peptides in innate immune responses. AB - Antimicrobial peptides (AMPs) are ancient effector molecules in the innate immune response of eukaryotes. These peptides are important for the antimicrobial efficacy of phagocytes and for the innate immune response mounted by epithelia of humans and other mammals. AMPs are generated either by de novo synthesis or by proteolytic cleavage from antimicrobially inactive proproteins. Studies of human diseases and animal studies have given important clues to the in vivo role of AMPs. It is now evident that dysregulation of the generation of AMPs in innate immune responses plays a role in certain diseases like Crohn's disease and atopic dermatitis. AMPs are attractive candidates for development of novel antibiotics due to their in vivo activity profile and some peptides may serve as templates for further drug development. PMID- 18511857 TI - Complement: an efficient sword of innate immunity. AB - Complement is vital for protecting individuals against pathogens and any disturbance of homeostasis associated with appearance of foreign antigens. Four antenna molecules seek for putative danger and subsequently start three activation pathways to eliminate the hostile triggering signal. To achieve this mission the complement arsenal contains soluble plasma factors as well as membrane-bound receptor molecules. Fulfilling a broad spectrum of biological functions, complement participates to construct and orchestrate an immunological network with extensive links to other elements of innate immunity, but also to its younger brother, the adaptive immune system. The body generously supports the complement activity with a high level of complement production; not only the liver as 'the capital of complement expression' but also decentralized synthesis sites guarantee its all-over presence. On the other hand, it is of fundamental interest for the organism to limit this powerful immunological regiment by establishing a tight surveillance composed of redundantly acting regulator molecules. To find the appropriate dimension of complement activity is critical, as shown by the spectrum of diseases associated with an excess or a lack. Numerous therapeutic approaches aim to correct such an imbalance and to re establish the antimicrobial capacity of complement without induction of chronic inflammation and autoimmunity. PMID- 18511858 TI - Antibacterial chemokines--actors in both innate and adaptive immunity. AB - Several antibacterial proteins and peptides of the human innate immune system have additional roles in the regulation of adaptive immune responses. Among peptides with innate and adaptive immune functions are chemokines, a family of structurally related peptides with conserved amino-terminal motifs. Chemokines regulate leukocyte trafficking during both health and disease. In recent years, some chemokines have been shown to exert direct antibacterial activity. On the other hand, several granulebound antibacterial proteins of granulocytes, and epithelium-expressed antibacterial polypeptides, possess chemotactic activity and stimulate cells of the adaptive immune system. It is likely that during evolution, some antimicrobial peptides and proteins of innate immunity have diverged to coordinate the actions of the innate immune system and the evolutionary younger, adaptive immunity. This review aims to describe antibacterial chemokines and antibacterial peptides possessing chemotactic activity, biologic properties that link innate and adaptive immunity. PMID- 18511859 TI - The role of neutrophils and monocytes in innate immunity. AB - Polymorphonuclear neutrophils (PMNs) and monocyte/macrophages (MMs) are professional phagocytic cells that are able to phagocytose and destroy infectious agents. Therefore, they are key anti-infectious actors in host defense but can mediate tissue damages. In addition, it is now clear that the role of these cells goes far beyond phagocytosis and pathogen killing. PMNs and MMs are essential cells for immunity, absolutely required to build and modulate the innate response. The respective roles of PMNs and MMs in the inflammatory process are discussed: their common features and their differences are reviewed, both in terms of origins and functions with special emphasis on novel concepts about neutrophil survival and resolution of inflammation. The recognition and the subsequent engulfment of apoptotic PMNs by macrophages is a key event of the resolution of inflammation, which can be associated with autoimmunity or inflammatory diseases. During the past years, significant efforts have been made to dissect the molecular mechanisms governing phagocytosis and pathogen killing. Although these effector functions are crucial, more work has to be done to understand the respective role of PMNs and MMs to regulate and inhibit the inflammatory process as well as the immune response. This might be the future challenge for the next years in phagocyte research and this will presumably open new avenues of research in the modulation of inflammation. PMID- 18511860 TI - Innate immune functions of the airway epithelium. AB - The epithelium of the respiratory tract forms a large surface area that maintains intimate contact with the environment. Through the act of breathing, this mucosal surface encounters an array of pathogens and toxic particulates. In response to these challenges many strategies have evolved to protect the host. These include the barrier functions of the epithelium, cough, mucociliary clearance, resident professional phagocytes, and the secretion of a number of proteins and peptides with host defense functions. Thus, the surface and submucosal gland epithelium of the conducting airways is a constitutive primary participant in innate immunity. In addition, this tissue may serve the function of a secondary amplifier of innate immune responses following neurohumoral input, stimulation with cytokines from cells such as alveolar macrophages, or engagement of pattern recognition receptors. Here, we provide an overview of the airway epithelium's role in pulmonary innate immunity, especially in the context of bacterial and viral infections, emphasizing findings from human cells and selected animal models. We also provide examples of human disease states caused by impaired epithelial defenses in the lung. PMID- 18511861 TI - Oxidative innate immune defenses by Nox/Duox family NADPH oxidases. AB - The importance of reactive oxygen species (ROS) in innate immunity was first recognized in professional phagocytes undergoing a 'respiratory burst'upon activation. This robust oxygen consumption is related to a superoxide-generating enzyme, the phagocytic NADPH oxidase (Nox2-based or phox). The oxidase is essential for microbial killing, since patients lacking a functional oxidase suffer from enhanced susceptibility to microbial infections. ROS derived from superoxide attack bacteria in the isolated niche of the neutrophil phagosome. The oxidase is electrogenic, alters ion currents across membranes, induces apoptosis, regulates cytokine production, influences gene expression, and promotes formation of extracellular traps. Recently, new homologues of Nox2 were discovered establishing the Nox family of NADPH oxidases that encompasses seven members. Nox1 is highly expressed in the colon epithelium, and can be induced by LPS or IFN- gamma. Nox4 was implicated in innate immunity since LPS induces Nox4 dependent ROS generation. Duox1 and Duox2 localize to the apical plasma membrane of epithelial cells in major airways, salivary glands, and the gastrointestinal tract, and provide extracellular hydrogen peroxide to lactoperoxidase to produce antimicrobial hypothiocyanite ions. Th1 and Th2 cytokines regulate expression of dual oxidases in human airways and may thereby act in host defense or in proinflammatory responses. PMID- 18511862 TI - Aging and impairment of innate immunity. AB - As we age, it is common for certain phenotypic changes to arise within the population. A number of observations have led scientists to believe that these changes result from an accumulation of cellular defects over time. With enough cell damage, tissue function is compromised and the risk for disease escalates. More importantly, when these defects arise in cells of the innate immune system, the body can no longer defend itself against a variety of pathologies. The main culprit for cellular damage seen with age is thought to be reactive oxygen and nitrogen species produced from endogenous metabolic pathways. To determine how an individual will age, it is thus important to consider all of the factors involved in both the production of and the response to oxidative stress. These factors include genetics, lifestyle, environment, and gender. Understanding the mechanisms of aging can allow us to develop strategies for overcoming the negative aspects of this process and ultimately to help individuals age more gracefully. PMID- 18511863 TI - Effects of sex steroids on the neurotransmitter-specific aromatic amino acids phenylalanine, tyrosine, and tryptophan in transsexual subjects. AB - BACKGROUND/AIMS: Phenylalanine, tyrosine and tryptophan are essential aromatic amino acids and precursors of dopamine, norepinephrine, epinephrine and serotonin. The aim of this study was to assess whether sex steroids affect plasma levels of these aromatic amino acids. METHODS: 15 male-to-female (M-F) transsexuals were treated with 100 microg/day ethinyl estradiol and 100 mg/day cyproterone acetate, and 14 female-to-male (F-M) transsexuals were treated with testosterone esters 250 mg i.m. per 2 weeks. Plasma levels of hormones and amino acids were measured at baseline and after 4 and 12 months of cross-sex hormone administration, and analyzed by general linear models for repeated measures. RESULTS: Plasma phenylalanine decreased by 7.5% (SD 3.0; p = 0.01); tyrosine by 18.3% (SD 4.6; p < 0.001), and tryptophan by 7.8% (SD 4.7; p = 0.03) after 12 months of estrogen + anti-androgen administration to M-F transsexuals. Administration of testosterone in F-M transsexuals did not induce significant changes in plasma levels of phenylalanine and tyrosine, but increased plasma tryptophan by 18.2% (SD 20.6; p = 0.001). CONCLUSION: Estrogens and anti androgens reduce circulating levels of phenylalanine, tyrosine, and tryptophan in men, whereas testosterone administration increases plasma tryptophan levels in women. Sex steroids may influence the availability of neurotransmitter precursors. PMID- 18511864 TI - Increased values of mean platelet volume and platelet size deviation width may provide a safe positive diagnosis of idiopathic thrombocytopenic purpura. AB - INTRODUCTION: It has been shown recently that platelet indices like mean platelet volume (MPV), platelet size deviation width (PDW) and platelet-to-large-cell ratio (P-LCR) are helpful in the discrimination between hyperdestructive thrombocytopenia like idiopathic thrombocytopenic purpura (ITP) and hypoproductive thrombocytopenia (HT). The aim of the study is to assess the reliability of these indices in the differentiation of ITP from other thrombocytopenias. METHODS: We recruited 134 thrombocytopenic patients (69 men, 65 women) who were divided into two groups according to the underlying disease: group I (n = 63) included ITP patients, whereas group II (n = 71) included patients with HT due to myelosuppression secondary to chemotherapy for hematological malignancies. Platelet indices were derived from a Sysmex automated cell counter. Sensitivity, specificity, positive prognostic value, negative prognostic value, efficiency and Youden index were calculated. RESULTS: Concerning MPV and PDW indices, sensitivity, specificity, positive prognostic value, negative prognostic value, efficiency and Youden index were 100% for the diagnosis of ITP. On the contrary, the values for P-LCR were significantly lower. CONCLUSIONS: MPV and PDW can be safely relied on for a positive diagnosis of ITP. MPV and PDW were superior to P-LCR. PMID- 18511865 TI - Neurological symptoms in aortic dissection: a challenge for neurologists. AB - Typically, aortic dissection has to be considered in patients with acute thoracic or abdominal pain and accompanying cardiovascular symptoms. Due to these clinical symptoms, neurologists have not been involved in the routine emergency management of aortic dissection. However, transient or permanent neurological symptoms at onset of aortic dissection are not only frequent (17-40% of the patients), but often dramatic and may mask the underlying condition. Especially in pain-free dissection (which occurs in 5-15%) with predominant neurological symptoms diagnosis of aortic dissection can be difficult and delayed. Affecting the outflow of supra-aortal, spinal as well as extremity arteries leads to a variety of neurological symptoms including disturbances of central or peripheral nervous system. Thrombolysis as an emergency stroke therapy without considering aortic dissection may be life-threatening for these patients. Routine chest X-ray and being alert to physical examination findings such as hypotension, asymmetrical pulses or cardiac murmur may reduce risk of delayed diagnosis or misdiagnosis. Neurological symptoms at onset or in the postoperative course of aortic dissection are not necessarily associated with increased mortality. PMID- 18511866 TI - Cerebral hemodynamics and oxygen metabolism in patients with moyamoya syndrome associated with atherosclerotic steno-occlusive arterial lesions. AB - BACKGROUND: Patients with major cerebral artery steno-occlusion and the formation of the moyamoya-like vessels associated with some other disorders have been distinguished from moyamoya disease and classified as moyamoya syndrome. The hemodynamic and metabolic backgrounds of the moyamoya syndrome associated with atherosclerosis have not yet been investigated. We aimed to elucidate the hemodynamic and metabolic characteristics associated with the development of basal moyamoya-like vessels in moyamoya syndrome with atherosclerosis. METHODS: Twenty-one patients with chronic unilateral atherosclerotic steno-occlusive lesions of the internal carotid artery or middle cerebral artery (MCA) were enrolled in the study. Based on the angiographic findings, the patients were classified into 2 groups: the moyamoya syndrome group (n = 7) and the non moyamoya-syndrome group (n = 14). We conducted angiographic evaluations of the extent of the development of basal moyamoya-like vessels in the moyamoya syndrome group. The cerebral blood flow, cerebral metabolic rate of oxygen, oxygen extraction fraction (OEF) and cerebral blood volume were measured using PET in the ipsilateral MCA area in the patients and in normal controls (n = 6). RESULTS: The OEF in the ipsilateral MCA area, except in the basal ganglia, was significantly higher in the moyamoya syndrome group than in the non-moyamoya syndrome group (p < 0.001). The extent of the development of basal moyamoya-like vessels was closely correlated with the elevation of the OEF (r > 0.999, p < 0.001). CONCLUSION: The basal moyamoya-like vessels are evidence of misery perfusion in patients with unilateral chronic atherosclerotic steno-occlusive lesions of major cerebral artery trunks. PMID- 18511867 TI - Quantification of the probability of reaching mobility independence at discharge from a rehabilitation hospital in nonwalking early ischemic stroke patients: a multivariate study. AB - BACKGROUND: This study was designed to quantify the probability of recovery of mobility in admission nonwalking stroke survivors. METHODS: We evaluated 437 of 500 consecutive patients admitted for sequelae of first ischemic stroke within the first month. We performed several logistic regressions using mobility status at discharge (independence in stair climbing; walking outside and inside, without aid or supervision; walking with cane or other aid, or need for wheelchair) as dependent variable, and several independent variables, including stratification of patients according to their Barthel Index (BI) score into 6 classes (< or =10; 11-20; 21-30; 31-40; 41-50; 51-60). RESULTS: At discharge, 4.58% of patients were independent in stair climbing, 8.70% were able to walk outside, 14.41% to walk inside, and 27.46% to walk with cane or other aid, while 44.85% remained in wheelchair. Very low BI scores at admission were associated with a high risk of need for wheelchair, whereas patients with BI score 51-60 showed a high probability to reach independence in stair climbing (OR = 5.60). Age, severity of neurological impairment, global aphasia, unilateral spatial neglect, male gender and vocational status also played a prognostic role. CONCLUSIONS: The probability of potential mobility recovery can be quantified at admission with better accuracy for independence in stair climbing and walking outside without any aid (percentages correctly predicted 95.4 and 91.8%, respectively). Stratification of BI score may be useful to better quantify the risk for each patient. PMID- 18511868 TI - Vessel wall contrast enhancement: a diagnostic sign of cerebral vasculitis. AB - PURPOSE: Inflammatory stenoses of cerebral arteries cause stroke in patients with florid vasculitis. However, diagnosis is often difficult even with digital subtraction angiography (DSA) and biopsy. The purpose of this study was to establish the value of contrast-enhanced MRI, proven to be sensitive to extradural arteritis, for the identification of intracranial vessel wall inflammation. PATIENTS AND METHODS: Twenty-seven patients with a diagnosis of cerebral vasculitis affecting large brain vessels were retrieved from the files: 8 children (2-10 years, 7 female, 1 male) and 19 adults (16-76 years, 10 female, 9 male). Diagnosis was based on histological or serological proof of vasculitis or on clinical and imaging criteria. All MRI examinations included diffusion weighted imaging, time-of-flight magnetic resonance angiography (TOF-MRA) and contrast-enhanced scans. MRI scans were assessed for the presence of ischemic brain lesions, arterial stenoses, vessel wall thickening and contrast uptake. RESULTS: Ischemic changes of the brain tissue were seen in 24/27 patients and restricted diffusion suggestive of recent ischemia in 17/27; 25/27 patients had uni- or multifocal stenoses of intracranial arteries on TOF-MRA and 5/6 had stenoses on DSA. Vessel wall thickening was identified in 25/27, wall enhancement in 23/27 patients. CONCLUSION: Wall thickening and intramural contrast uptake are frequent findings in patients with active cerebral vasculitis affecting large brain arteries. Further prospective studies are required to determine the specificity of this finding. PMID- 18511869 TI - Inhibitory effects of eicosapentaenoic acid on chronic cerebral vasospasm after subarachnoid hemorrhage: possible involvement of a sphingosylphosphorylcholine rho-kinase pathway. AB - BACKGROUND AND PURPOSE: Rho-kinase (ROK)-mediated Ca2+ sensitization of vascular smooth muscle (VSM) contraction plays a pivotal role in cerebral vasospasm (CV). We previously demonstrated that sphingosylphosphorylcholine (SPC) induces Ca2+ sensitization through sequential activation of the Src family protein tyrosine kinases (Src-PTKs) and ROK in vitro, and that Ca2+ sensitization is inhibited by eicosapentaenoic acid (EPA) through the selective inactivation of Src-PTK. In this study, we examined whether SPC induced CV in vivo, and, if it did, whether EPA would inhibit CV, as induced by SPC or in an in vivo model of subarachnoid hemorrhage (SAH). METHODS: Changes in the diameter of the canine basilar artery were investigated by angiography after administering SPC into the cisterna magna. Then, Y27632, a specific Rho-kinase inhibitor, or EPA was injected intracisternally and the effects of both agents were investigated. In another experiment using a single-hemorrhage model, Y27632 or EPA was injected on day 7 after SAH and the changes in the diameter of the canine basilar artery were investigated. RESULTS: At cerebrospinal fluid concentrations of 100 and 300 micromol/l, SPC induced severe vasoconstriction (maximum vasoconstriction by SPC (100 micromol/l): 61.8 +/- 8.2%), which was markedly reversed by Y27632 (96.3 +/- 4.4%) or EPA (92.6 +/- 12.8%). SAH caused severe vasospasm on day 7 (67.6 +/- 7.8%), which was significantly blocked by Y27632 (95.5 +/- 10.6%) or EPA (90.0 +/ 4.4%). CONCLUSIONS: SPC is a novel mediator of ROK-induced CV in vivo. The inhibition of CV induced by SPC or after SAH by EPA suggests beneficial roles of EPA in the treatment of CV. Our findings are compatible with the notion that the SPC-ROK pathway may be involved in CV. PMID- 18511870 TI - The complications of cardioembolic stroke: lessons from the VISTA database. AB - BACKGROUND: Cardioembolic stroke (CES) accounts for 20-30% of ischemic strokes. We attempted to describe the complications of CES. METHODS: We analyzed a cohort of CES patients from the VISTA database who were not given thrombolytics. RESULTS: 397 patients were included. Recurrent stroke occurred in 2.5% (10/397); stroke progression in 7.8% (31/397); systemic bleeding in 3% (12/397), and hemorrhagic transformation (HT) in 1.5% (6/397). A bimodal temporal distribution was observed for HT. The mortality rate was 103/397 (25.9%) and was associated with stroke progression and recurrence. CONCLUSIONS: HT is common in CES patients and presents in a bimodal distribution. Stroke progression is the most frequent serious complication. Further study is indicated to identify predictors of stroke progression. PMID- 18511871 TI - Towards a basic endoscopic assessment of swallowing in acute stroke - development and evaluation of a simple dysphagia score. AB - BACKGROUND AND PURPOSE: Dysphagia is an important complication of acute stroke with a reported incidence of up to 76%. The purpose of this investigation was to develop and to evaluate an endoscopic scoring system which has the potential to guide dysphagia management in acute stroke patients. METHODS: A fiberoptic endoscopic evaluation of swallowing (FEES) was carried out in 100 patients within 72 h of stroke onset. During endoscopic examination, the secretion status was evaluated, and the patient was successively given standard volumes of puree consistency, liquids and soft solid food. Penetration and aspiration was assessed using a 5-point scale. RESULTS: Nearly 80% of patients showed penetration or aspiration during FEES. These events occurred more frequently with liquids and soft solid food than with puree. Penetration or aspiration at any stage of examination predicted failure at the subsequent food consistency. Furthermore, key findings of endoscopic evaluation were predictive of the need for later orotracheal intubation. Based on these observations a score was developed that grades stroke-related dysphagia according to the risk of penetration or aspiration of the different food consistencies tested. Assessment of interrater reliability by means of 25 additional endoscopic examinations resulted in excellent agreement between three investigators, reflected by a kappa coefficient of 0.89. CONCLUSION: Stroke-related dysphagia may effectively be graded using a simple endoscopic scoring system. The usefulness of this screening protocol with respect to patient outcome and intercurrent complications has to be studied in prospective clinical trials. PMID- 18511872 TI - Meta-analysis of genetic studies from journals published in China of ischemic stroke in the Han Chinese population. AB - BACKGROUND: The aim of this study was to confirm the nature and number of genes contributing to stroke risk and qualify the genetic risk of each susceptibility gene in the Han Chinese population. METHODS: After collecting all case-control studies related to DNA polymorphism of any candidate gene for ischemic stroke in Han Chinese, strict selection criteria and exclusion criteria were determined and different effect models were used according to the difference in heterogeneity. Meta-analyses were carried out by Revman 4.0 software and the publication bias was further evaluated through calculation of fail-safe numbers in the included gene polymorphisms. RESULTS: Seventy-six studies were included in the meta analyses which were all published in mainland China and referred to 6 candidate genes and 7 polymorphisms. Among the gene polymorphisms tested in the study, association of gene polymorphisms with increasing risk of ischemic stroke was confirmed in 6 polymorphisms including angiotensin-converting enzyme insertion/deletion (ACE I/D; OR = 1.87, 95% CI = 1.45-2.42), methylenetetrahydrofolate reductase (MTHFR) C677T (OR = 1.55, 95% CI = 1.26 1.90), plasminogen activator inhibitor 1 (PAI-1) 4G/5G (OR = 1.79, 95% CI = 1.20 2.67), beta-fibrinogen (beta-Fg) -455A/G (OR = 1.48, 95% CI = 1.14-1.92), beta-Fg -148T/C (OR = 1.72, 95% CI = 1.42-2.07), apolipoprotein E (ApoE) epsilon2-4 (OR = 2.39, 95% CI = 1.94-2.95). Because of the obvious publication bias, the association between paraoxonase 1 (PON-1) polymorphisms and stroke risk was not established although the OR of the meta-analysis suggested a positive result (OR = 1.14, 95% CI = 1.01-1.35). CONCLUSIONS: ACE D/I, MTHFR C677T, beta-Fg -455A/G, beta-Fg -148T/C, PAI-1 4G/5G, and ApoE epsilon2-4 were associated with risk of ischemic stroke in Han Chinese. PMID- 18511873 TI - Antiplatelet cilostazol is beneficial in diabetic and/or hypertensive ischemic stroke patients. Subgroup analysis of the cilostazol stroke prevention study. AB - BACKGROUND AND PURPOSE: Although antiplatelets are known to be effective for secondary prevention of cerebral infarction, the number needed to treat is rather large and the effects in stroke patients with complications such as hypertension or diabetes are inadequately defined. This study was conducted to examine the effect of such complications on recurrence of cerebral infarction, and to assess the effect of cilostazol, an antiplatelet agent, in these high-risk subjects. METHODS: A post hoc subgroup analysis of the already reported Cilostazol Stroke Prevention Study, which was a placebo-controlled double-blind trial, has been carried out to clarify the influence of various complications on recurrence in the placebo group and the effects of cilostazol in 1,095 patients with noncardioembolic ischemic cerebrovascular disease. Treatment continued for an average of 1.8 +/- 1.3 years (maximum 4.8 years). RESULTS: The recurrence rate of the diabetic stroke patients was significantly higher compared with the nondiabetics in the placebo group (9.4 vs. 4.7%/year, p = 0.01). Furthermore, our study showed that the relative risk reduction (RRR) for recurrence of infarction was 41.7% with cilostazol. This treatment provided a significant benefit in patients with lacunar infarction (RRR 43.4%, p = 0.04), with diabetes (RRR 64.4%, p = 0.008), or with hypertension (RRR 58.0%, p = 0.003). CONCLUSIONS: Diabetic patients are particularly at risk for recurrence of cerebral infarction. Cilostazol is useful for the prevention of the recurrence of vascular events in patients with lacunar infarction, and is probably effective in high-risk patients with diabetes and/or hypertension. PMID- 18511874 TI - Oxidative stress and asymmetric dimethylarginine are associated with cardiovascular complications in hemodialysis patients: improvements by L-arginine intake. AB - BACKGROUND/AIM: High incidence of cardiovascular disease (CVD) in chronic kidney disease (CKD) patients is a result of an interlaced relation between oxidative stress, endothelial dysfunction (ED) and inflammation. This study tries to investigate the development of these processes in CKD patients receiving conservative treatment or on hemodialysis (HD). We also examined the modulating effect of oral L-arginine in HD patients having CVD. METHODS: The study included 12 healthy volunteers and 63 renal patients divided into 15 renal impairment, 18 HD free of CVD, and 30 HD suffering from CVD (HD+CVD). Of the latter, 15 patients were given oral L-arginine (15 g/day, 5 g t.i.d.) for 1 month. Blood levels of asymmetric dimethylarginine (ADMA), malondialdehyde (MDA), and homocysteine and myeloperoxidase activity (MPO) were estimated. RESULTS: ADMA, MDA and homocysteine were significantly elevated in renal impairment group. HD and HD+CVD patients experienced higher levels, along with high MPO activity. Significant reduction by 21, 46, 11, and 26%, respectively, in the aforementioned parameters was observed in HD+CVD patients following L-arginine intake. CONCLUSION: We recommend considering ADMA, MDA, homocysteine and MPO as potentially important cardiovascular risk factors in CKD patients, and focus the attention to the cardiovascular advantages of L-arginine in these patients. PMID- 18511875 TI - An ultrasensitive new DNA microarray chip provides gene expression profiles for preoperative esophageal cancer biopsies without RNA amplification. AB - OBJECTIVES: Gene expression profiling using pretreatment biopsies has been limited due to their small sample sizes. This study evaluated the usefulness of an ultrasensitive new DNA microarray chip, which has a unique array structure, for the clinical diagnosis of esophageal cancer using preoperative biopsies. METHODS: Paired cancer and normal esophageal epithelial tissues from 56 patients who underwent esophagectomy and from 48 patients who underwent preoperative endoscopy were studied. Among 2 feature gene sets selected by a reference DNA chip discriminating malignant status of samples, 20 feature genes were selected for the development of the new DNA chip. The new DNA chip was hybridized with 0.1 mug of total RNA per slide without RNA amplification. RESULTS: Twenty feature genes, including RRM-2 and XRCC-3, for the new DNA chip could discriminate cancer from noncancer at a 95.2% rate of accuracy in 42 biopsies (sensitivity 95.7%, specificity 94.7%). A receiver operating characteristic (ROC) curve analysis showed that the area under ROC curve for the prediction was 0.966. CONCLUSIONS: The gene expression profiles from the preoperative biopsies could diagnose esophageal cancer accurately, using the ultrasensitive DNA chip without RNA amplification. This new DNA chip technology might contribute further to the development of customized therapeutic strategies for various cancer patients. PMID- 18511876 TI - Association of E1AF mRNA expression with tumor progression and matrilysin in human rectal cancer. AB - OBJECTIVE: To examine E1AF mRNA expression and to determine whether it is correlated with tumor progression and matrilysin in human rectal cancer. METHODS: Real-time RT-PCR was used to determine E1AF and matrilysin expression in 100 matched rectal cancers and normal tissues. RESULTS: Among the 100 rectal cancers, 69 cases of E1AF mRNA overexpression were observed. E1AF mRNA overexpression correlated well with matrilysin. In carcinomas, E1AF mRNA overexpression correlated significantly with depth of invasion, lymph node metastasis, venous involvement and advanced pTNM stage. CONCLUSIONS: E1AF was correlated significantly with tumor progression of human rectal cancer and may be an important factor in rectal cancer progression. PMID- 18511877 TI - Immunohistochemical study of glypican 3 in thyroid cancer. AB - In 123 patients with thyroid cancer, expression of glypican 3 (GPC3) was immunohistochemically investigated in tissue samples and the biological significance of GPC3 in thyroid cancer was examined. GPC3 was scarcely expressed in the normal thyroid gland, but was dramatically enhanced in certain types of cancers: 100% in follicular carcinoma (20/20 cases) and 70% in papillary carcinoma (48/69 cases). Expression of GPC3 in follicular carcinoma was significantly higher than that of follicular adenoma (p < 0.0019). In contrast, GPC 3 was not expressed in 17 cases of anaplastic carcinoma. A high expression of GPC3 mRNA was confirmed in cancer lesions, which were strongly positive for immunohistochemical staining. In 69 cases of papillary carcinoma, GPC3 was expressed at an early stage, suggesting that GPC3 expression in thyroid cancer is an early event in developing papillary carcinoma. Further studies are required to determine biological functions and molecular mechanisms underlying the upregulation of GPC3 in thyroid cancer. PMID- 18511878 TI - Physiological function of NbRanBP1 in Nicotiana benthamiana. AB - This study addresses the physiological functions of the Ran-binding protein homolog NbRanBP1 in Nicotiana benthamiana. Virus-induced gene silencing (VIGS) of NbRanBP1 caused stunted growth, leaf yellowing, and abnormal leaf morphology. The NbRanBP1 gene was constitutively expressed in diverse tissues and an NbRanBP1:GFP fusion protein was primarily localized to the nuclear rim and the cytosol. BiFC analysis revealed in vivo interaction between NbRanBP1 and NbRan1 in the nuclear envelope and the cytosol. Depletion of NbRanBP1 or NbRan1 reduced nuclear accumulation of a NbBTF3:GFP marker protein. In the later stages of development, NbRanBP1 VIGS plants showed stress responses such as reduced mitochondrial membrane potential, excessive production of reactive oxygen species, and induction of defense-related genes. The molecular role of RanBP1 in plants is discussed in comparison with RanBP1 function in yeast and mammals. PMID- 18511879 TI - Involvement of corticotropin-releasing factor receptor 2 beta in differentiation of dopaminergic MN9D cells. AB - Corticotropin releasing factor (CRF) mediates various responses to stress through CRF receptors 1 and 2. CRF receptor 2 has two forms, 2alpha and 2beta each of which appears to have distinct roles. Here we used dopaminergic neuron-derived MN9D cells to investigate the function of CRF receptor 2 in dopamine neurons. We found that n-butyrate, a histone deacetylase inhibitor, induced MN9D cell differentiation and increased gene expression of all CRF receptors. CRF receptor 2beta was minimally expressed in MN9D cells; however, its expression dramatically increased during differentiation. CRF receptor 2beta expression levels appeared to correlate with neurite outgrowth, suggesting CRF receptor 2beta involvement in neuronal differentiation. To validate this statement, we made a CRF receptor 2beta-overexpressing MN9D/CRFR2 beta stable cell line. This cell line showed robust neurite outgrowth and GAP43 overexpression, together with MEK and ERK activation, suggesting MN9D cell neuronal differentiation. From these results, we conclude that CRF receptor 2beta plays an important role in MN9D cell differentiation by activating the MEK/ERK signaling pathway. PMID- 18511880 TI - Forced expression of HoxB4 enhances hematopoietic differentiation by human embryonic stem cells. AB - HoxB4 has been shown to enhance hematopoietic engraftment by hematopoietic stem cells (HSC) from differentiating mouse embryonic stem cell (mESC) cultures. Here we examined the effect of ectopic expression of HoxB4 in differentiated human embryonic stem cells (hESCs). Stable HoxB4-expressing hESCs were established by lentiviral transduction, and the forced expression of HoxB4 did not affect stem cell features. HoxB4-expressing hESC-derived CD34+ cells generated higher numbers of erythroid and blast-like colonies than controls. The number of CD34+ cells increased but CD45+ and KDR+ cell numbers were not significantly affected. When the hESC derived CD34+ cells were transplanted into NOD/SCID beta 2m-/- mice, the ectopic expression of HoxB4 did not alter their repopulating capacity. Our findings show that overexpression of HoxB4 in differentiating hESCs increases hematopoietic colony formation and hematopoietic cell formation in vitro, but does not affect in vivo repopulation in adult mice hosts. PMID- 18511881 TI - Association of A/T rich microsatellites with responses to artificial selection for larval developmental duration in the silkworm Bombyx mori. AB - Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) (29.428 (+/-) 0.723 days) and shorter larval duration (SLD) (22.573 (+/-) 0.839 days) lines from a base, inbred population of Nistari (larval span of 23.143 (+/-) 0.35 days). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 (CA7) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the F2 generation of the LLD x SLD lines, the alleles of Bmsat106 and UBC8731074bp segregated in a 1:1 ratio and the loci were present only in the LLD individuals. UBC873823bp was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the UBC8731074bp and UBC873823bp loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the UBC8731074bp and UBC873823bp loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the UBC8731074bp sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed approximately 49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics. PMID- 18511882 TI - Soluble expression of recombinant olive flounder hepcidin I using a novel secretion enhancer. AB - Expression of olive flounder hepcidin I (HepI) fused with truncated OmpA signal peptides (OmpASP(tr)) as directional signals does not produce soluble fusion proteins. However, by inserting amino acid segments (xxx) varying in pI and hydrophobicity/hydrophilicity into a leader sequence containing a truncated OmpASP (OmpASP(tr)) and a factor Xa cleavage site (Xa) [OmpASP(tr)|(xxx)|Xa], we were able in some cases to express soluble recombinant HepI. Soluble expression of the recombinant protein strongly correlated with (xxx) insertions of high pI and hydrophilicity. Therefore, we modified the OmpASP(tr)|(xxx)|Xa sequence by inserting Arg and Lys into (xxx) to increase the hydrophilicity of the signal peptide region. These modifications enhanced the expression of soluble recombinant HepI. Hydropathic profile analysis of the OmpASP(tr)|(xxx)|Xa| HepI fusion proteins revealed that the transmembrane-like domains derived from the OmpASP(tr)|(xxx)|Xa sequence were larger than the internal positively charged domain native to HepI. It should therefore be possible to overcome the obstacle of internal positively charged domains to obtain soluble expression of recombinant proteins by monitoring the hydrophilicity and hydropathic profile of the signal peptide region using a computer program. PMID- 18511883 TI - Training molecularly enabled field biologists to understand organism-level gene function. AB - A gene's influence on an organism's Darwinian fitness ultimately determines whether it will be lost, maintained or modified by natural selection, yet biologists have few gene expression systems in which to measure whole-organism gene function. In the Department of Molecular Ecology at the Max Planck Institute for Chemical Ecology we are training "molecularly enabled field biologists" to use transformed plants silenced in the expression of environmentally regulated genes and the plant's native habitats as "laboratories." Research done in these natural laboratories will, we hope, increase our understanding of the function of genes at the level of the organism. Examples of the role of threonine deaminase and RNA-directed RNA polymerases illustrate the process. PMID- 18511884 TI - Suppression of arsenic trioxide-induced apoptosis in HeLa cells by N acetylcysteine. AB - Arsenic trioxide (ATO) can affect many biological functions such as apoptosis and differentiation in various cells. We investigated the involvement of ROS and GSH in ATO-induced HeLa cell death using ROS scavengers, especially N-acetylcysteine (NAC). ATO increased intracellular O(2)(*-) levels and reduced intracellular GSH content. The ROS scavengers, Tempol, Tiron and Trimetazidine, did not significantly reduce levels of ROS or GSH depletion in ATO-treated HeLa cells. Nor did they reduce the apoptosis induced by ATO. In contrast, treatment with NAC reduced ROS levels and GSH depletion in the ATO-treated HeLa cells and prevented ATO-induced apoptosis. Treatment with exogenous SOD and catalase reduced the depletion of GSH content in ATO-treated cells. Catalase strongly protected the cells from ATO-induced apoptosis. In addition, treatment with SOD, catalase and NAC slightly inhibited the G1 phase accumulation induced by ATO. In conclusion, NAC protects HeLa cells from apoptosis induced by ATO by up-regulating intracellular GSH content and partially reducing the production of O(2)(*-). PMID- 18511885 TI - Raloxifene, a selective estrogen receptor modulator, inhibits lipopolysaccharide induced nitric oxide production by inhibiting the phosphatidylinositol 3 kinase/Akt/nuclear factor-kappa B pathway in RAW264.7 macrophage cells. AB - We here demonstrate an anti-inflammatory action of raloxifene, a selective estrogen receptor modulator, in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells. Treatment with raloxifene at micromolar concentrations suppressed the production of nitric oxide (NO) by down-regulating expression of the inducible nitric oxide synthase (iNOS) gene in LPS-activated cells. The decreased expression of iNOS and subsequent reduction of NO were due to inhibition of nuclear translocation of transcription factor NF-kappaB. These effects were significantly inhibited by exposure to the phosphatidylinositol 3 kinase (PI 3-kinase) inhibitor, LY294002, or by expression of a dominant negative mutant of PI 3-kinase. In addition, pretreatment with raloxifene reduced LPS induced Akt phosphorylation as well as NF-kappa B DNA binding activity and NF kappa B-dependent reporter gene activity. Thus our findings indicate that raloxifene exerts its anti-inflammatory action in LPS-stimulated macrophages by blocking the PI 3-kinase-Akt-NF-kappa B signaling cascade, and eventually reduces expression of pro-inflammatory genes such as iNOS. PMID- 18511886 TI - A novel expression system for recombinant marine mussel adhesive protein Mefp1 using a truncated OmpA signal peptide. AB - To express an increased level of recombinant Mefp1 (marine mussel adhesive protein) in soluble form, we constructed expression vectors encoding truncated OmpA signal peptide-Mefp1 fusion proteins. OmpA signal peptide (OmpASP) is the 21 residue peptide fragment of the 23 residue OmpA signal sequence cleavable by signal peptidase I. We successfully produced increased levels of soluble recombinant Mefp1 (rMefp1) with various deletions of OmpASP, and found that the increased expression was caused by the increased pI of the N-terminus of the fusion proteins (> or = 10.55). All the OmpA signal peptide segments of 3-21 amino acids in length had the same pI value (10.55). Our results suggest that the pI value of the truncated OmpASP (OmpASP(tr)) play an important role in directional signaling for the fusion protein, but we found no evidence for the presence of a secretion enhancer in OmpASP. For practical applications, we increased the expression of soluble rMefp1 with OmpASP(tr) peptides as directional signals, and obtained rMefp1 with the native amino terminus (nN rMefp1) using an OmpASP(tr)| Xa leader sequence that contains the recognition site for Xa protease. PMID- 18511887 TI - The terminal and internal hairpin loops of the ctRNA of plasmid pJB01 play critical roles in regulating copy number. AB - The plasmid pJB01, a member of the pMV158 family isolated from Enterococcus faecium JC1, contains three open reading frames, copA, repB, and repC. Plasmids included in this family produce counter-transcribed RNA (ctRNA) that contributes to copy number control. The pJB01 ctRNA, a transcript which consists of 54 nucleotides (nts), is encoded on the opposite strand from the copA/repB intergenic region and partially overlaps an atypical ribosome binding site (ARBS) for repB. The ARBS is integrated by the two underlined conserved regions: 5' TTTTTGTNNNNTAANNNNN NNNNATG-3', and the ctRNA is complementary only to the 5' conserved sequence 5'-TTTTTGT-3'. This complementary sequence is located at a distance from the terminal loop of the ctRNA secondary structure. The ctRNA structure predicted by the mfold program suggests the possible generation of a terminal and an internal hairpin loop. The amount of in vitro translation product of repB mRNA was inversely proportional to the ctRNA concentration. Mutations in the terminal and internal hairpin loops of the ctRNA had inhibitory effects on its binding to the target mRNA. We propose that the intact structures of the terminal and internal hairpin loops, respectively, play important roles in forming the initial kissing and extending complexes between the ctRNA and target mRNA and that these regulate the copy number of this plasmid. PMID- 18511888 TI - Apoptotic cell death in TrkA-overexpressing cells: kinetic regulation of ERK phosphorylation and caspase-7 activation. AB - The TrkA tyrosine kinase is activated by autophosphorylation in response to NGF, and plays an important role in cell survival, differentiation, and apoptosis. To investigate its role in cell fate determination, we produced stable TrkA inducible SK-N-MC and U2OS cell lines using the Tet-On system. Interestingly, TrkA overexpression induced substantial cell death even in the absence of NGF, by stimulating ERK phosphorylation and caspase-7 activation leading to PARP cleavage. TrkA-mediated cell death was shown by the annexin-V binding assay to be, at least in part, apoptotic in both SK-N-MC and U2OS cells. Furthermore, the truncated form (p18) of Bax accumulated in the TrkA-induced cells, suggesting that TrkA induces mitochondria-mediated apoptosis. NGF treatment augmented the cell death induced by TrkA overexpression. This TrkA-induced cell death was blocked by the tyrosine kinase inhibitors, K-252a and GW441756. Moreover, TrkA overexpression inhibited long-term proliferation of both the neuronal SK-N-MC cells and the non-neuronal U2OS cells, suggesting a potential role of TrkA as a tumor suppressor. PMID- 18511889 TI - [Needs and expectations of adolescents affected by cancer]. PMID- 18511890 TI - [The treatment of pectus excavatum: results of a mininvasive surgical technique on the first 50 patients]. AB - AIM: Pectus excavatum is the commonest thoracic congenital malformation, but its treatment remains not well known. The authors present the results of the mini invasive repair at G. Gaslini Institute of Genoa, Italy. METHODS: Nuss mini invasive repair avoids anterior scars. The correction is achieved by the introduction under thoracoscopy of a retrosternal curve bar that is rotated by 180 degrees . Postoperatory pain is managed by an epidural catheter. In all the operated patients we evaluated the clinical pre-operatory parameters (spirometric, radiological and cardiological data), the surgical details and the results. RESULTS: Fifty patients were operated, 43 of them males, ranging from 7 and 22 years of age, with an average of 17 years of age. Only 8 of them were asymptomatic and required surgery for psychological reasons. The 74% presented some stress dyspnea. Some impairment in spirometric parameters were observed in 28% and mitral valve prolapse in 30%. The only significant intra-operative complication was a bleeding from a thoracic wall vessel that required a left emergency minimal thoracotomy. Postoperative complications were: 2 pneumothorax (drained for 24 hours), 2 transitory pulmonary atelectasis, 1 hemothorax in a patient with coagulation deficit, 3 wound problems (1 infection and 2 hematomas). The esthetical score after surgery, according to the patients, was 9.15 on average, in a scale from 1 to 10. None rated less than 7. The pain score with the same scale was rated 6.8 on average. CONCLUSION: The Nuss technique is safe and guarantees very satisfactory esthetical results. PMID- 18511891 TI - Association of physical fitness and body mass index in youth. AB - AIM: The aim of this study was to establish physical fitness (PF) levels in a school population of 11-18-year-old students and analyse differences according to body mass index (BMI) status in overweight METHODS: This is a cross-sectional study. The sample comprises 636 children and adolescents (mean age of 14.5+/-1.5 years), 288 boys (45.3%) and girls 347 (54.7%). Six tests from Fitness-gram battery were used as an objective measure of physical fitness. Overweight/ Obesity status was determined using age and sex adjusted cut-off points. RESULTS: Both girls and boys with obesity performed a significantly reduced number of tests in healthy fitness zone suggesting a decrease of performances in strength and cardiovascular fitness, from normal weight status to overweight and from overweight to obesity. Boys and girls with obesity are likely to be Under HFZ than normal weight. CONCLUSION: The results suggest that obese and overweight children have low PF level compared to normal weight peers. A large number of children with normal weight were identified as well as unfit. These data also showed that a low BMI level would significantly improve some PF component. PMID- 18511892 TI - [Conservative treatment of children constipation with Macrogol 4,000]. AB - AIM: Macrogol 4,000 is one of the new generation's osmotic laxatives. It is constituted by a heavy molecular weight polymer without additional salts. In most of patients Macrogol 4,000 shows its efficacy in 48 hours from the beginning of treatment. Daily evacuations has been reported after first week therapy with an improvement in quality of life. The aim of this observational study was to demonstrate the efficacy of Macrogol 4,000 in the treatment of constipation in children. METHODS: The effect of Macrogol 4,000 was assessed in 120 children affected by constipation treated with therapeutic doses of Macrogol 4,000. The study period ranged from September 2006 to March 2007. The data analysis was collected with the evaluations concerning clinical examination and the consultation of daily diaries assembled by parents, tracking stooling pattern; in case of loosing contacts with the patient, a telephonic survey has been carried out. The child with symptoms improvement, i.e. reduction of the pain during defecation, daily spontaneous evacuations and better stool consistence, were considered ''normal''. RESULTS: Of the 120 patients, 89 returned to the clinical follow-up, while the remaining 31, who missed the follow-up, has been contacted by telephone. To the final analysis, 103 patients (85.8%) have had a normalization of the clinical symptoms, 9 patients (7.5%) have refused Macrogol 4,000 for the unpleasant flavour, 8 patients (6.7%) did not have any improvement from the treatment. CONCLUSION: The results of this observational study suggest that Macrogol 4,000 could be very useful to treat pediatric patients with constipation, also those affected by neuronal intestinal displasia (NID) type B. PMID- 18511893 TI - [Newborn resuscitation: from necessity of continous practice to professional liability]. AB - The immediately postbirth extra uterine adaptation is the most important cause of death in the first two hours of life. In all risky cases, it is necessary to effect efficient and on time techniques of newborn resuscitation, because dubitation or delay may be very dangerous for the infant. In Italy courses of equipment in newborn resuscitation are regularly performed, but an excellent level of technique can be obtained only with continuous daily practice. Then, particularly in little hospitals where it is unusually necessary to act resuscitation on a newborn, courses of simulation for medical and nursering staff would be opportune to prevent neonatal handicap and to deal with the professional liability in the best way. The Italian current jurisprudence, in fact, has slowly confined the application of 2236 article of Civil code about professional liability in particularly difficult efforts. The Italian law asserts that a professional specialist is trained to be able resolve any type of problem among those of his specialistic competence, even if technically very difficult. It should be opportune to train health staff with practical exercises, in order to obtain complete technical skills in all neonatal centers. PMID- 18511894 TI - Adolescents and epilepsy: hormonal aspects. AB - Epilepsy is one of the most frequent disease in children, particularly in adolescents. Some types of epilepsy begin typically in the peripubertal period and many studies found changes in frequency or features of seizures in adolescents. This is probably related to sex hormone effects on neuronal excitability. Estrogens seem to decrease neuronal electric threshold with proconvulsant effects. On the contrary progesterone seems to have anticonvulsivant action, as well as testosterone. Changes in hormone serum levels that occur during pubertal period are probably related to modifications in seizure features and are involved in some types of epilepsy that begin typically in adolescence. PMID- 18511895 TI - [Priorix Tetra: a new combined vaccine against measles, rubella, mumps and varicella]. AB - Varicella is an infectious disease caused by a virus of the herpes virus family (VZV) affecting predominantly the pediatric age. Varicella is considered a mild disease, but in some cases, mainly in immunocompromised subjects, it can evolve towards complicated cases, even fatal. In immunocompetent subjects, however, serious complications can also arise, such as cutaneous bacterial superinfections, pneumonia, conjunctivitis or corneal infections and central nervous system (CNS) complications. The rate of hospitalization, even in developed countries, is high. Due to the epidemiological relevance of varicella, the availability of a live, attenuated, safe and effective vaccine, has prompted an international debate on the opportunity of extensive infant varicella vaccination, that could be overcome by the introduction of the combined tetravalent vaccines against measles, mumps, rubella and varicella (MMRV). Priorix Tetra, a new quadrivalent MMRV vaccine produced by GlaxoSmithKline, has demonstrated high immunogenicity, comparable to that of the separate vaccines (Priorix and Varilrix) in addition to a favorable tolerability and safety profile. This review summarizes the epidemiology of measles, mumps, rubella and varicella in Italy, encompassing the rationale for the introduction of varicella vaccination and describes the immunological, clinical and safety profile of Priorix Tetra. PMID- 18511896 TI - [Omega-3: from cod-liver oil to nutrigenomics]. AB - The leading role of cod-liver oil on rickets was a relevant factor in the knowledge of this disease. In 1922 the preventive and therapeutic value of cod liver oil and sunlight against rickets in young infants was confirmed. The seasonal variation in the incidence of rickets, the role of skin pigmentation, of diet and the fact that breast milk was not an adequate source of vitamin D were understood. The discovery of essential fatty acids omega-6 and omega-3 have shown that deficiencies, mainly of omega-3 long chain polyunsaturated fatty acids, result in visual and cognitive impairment and disturbances in mental functions in infants and also in cognitive function in adults, as fatty acids are beneficial to vascular health and may forestall cerebrovascular disease and thus dementia. An adequate ratio of n-6 and n-3 fatty acids may promote a healthier balance of eicosanoids, which would protect membrane function with a nutraceutical function. Dietary lipids not only influence the biophysical state of the cell membranes but, via direct and indirect routes, they also act on multiple pathways including signalling, gene and protein activities, protein modifications and they probably play important role in modulating protein aggregation. Significant advances have been made in understanding the relation between dietary factors and inflammation, which is a central component of many chronic diseases, including coronary artery disease, rheumatoid arthritis, cancer prevention. However, the identification of those who will or will not benefit from dietary intervention strategies remains a major obstacle. Adequate knowledge about how the responses depend on an individual's genetic background (nutrigenetic effects), the cumulative effects of food components on genetic expression profiles through nutrigenomics mechanism, may assist in identifying responders and non-responders. Thus, fish and fish oil consumption might encourage brain development and gene expression to brain maintenance during aging through nutrigenomic mechanism. PMID- 18511897 TI - Idiopathic infantile arterial calcification: case report. AB - Idiopathic infantile arterial calcificationI (IIAC) is a rare disorder characterized by calcium deposition in the internal elastica lamina of medium and large arteries and it has been defined in term of molecular genetics. It is usually fatal, approximately 85% of all patients die within the first months of life owing to ischemia of vital organs. Death from myocardial infarction usually occurs in the first 6 months. Calcification in a peripheral artery with electrocardiogram (ECG) changes of occlusive coronary artery disease suggests the diagnosis. It is caused by mutations in the ENPP1 gene localized on chromosome 6q22 and has OMIM number 208000. A case with fatal evolution at the end of the first month is presented. PMID- 18511898 TI - An unique case of Y-type urethral duplication associated with posterior urethral valve. AB - This case report is about a 4-year-old patient with a IIA2 ''Y-type'' duplication, with the accessory urethra arising from the anterior orthotopic urethra and exiting in perineal-scrotal position, in association with posterior urethral valves (PUV). Cystourethroscopy through the ventral urethra revealed a type III urethral valve (diaphragm) and this was fulgurated. Duplicated dorsal urethra was excised surgically. Postoperative period was uneventful. This case is unique, because it shows PUV in a child with a very rare type of ''Y-type'' duplication. The presence of PUV in patients with urethral duplication is probably not an incidental finding but, to date, embryology of this rare association is not known. PMID- 18511899 TI - Dandy-Walker syndrome together with occipital encephalocele. AB - Dandy-Walker malformation is an anomaly characterized by dysgenesis of the foramina of Magendie and Lushka in the upper 4(th) ventricle, hypoplasia of the cerebellar vermis and agenesis of the corpus callosum. Encephalocele is diagnosed from the calvarium defect, cerebrospinal fluid (CSF) and herniation of the meninges. It is the rarest neural tube defect. A 7 x 9 cm encephalocele was found on physical examination of a 6-day old baby boy patient. From cranial magnetic resonance, it was seen that the posterior fossa was enlarged with cysts and there was agenesis of the vermis. A connection was established between the ventricle and the development of cysts on the posterior fossa. These findings were evaluated as significant from the aspect of Dandy-Walker malformation. The extension of the bone defect in the left occipital area towards the posterior, and the cranio-caudal diameter reaching 9 cm was seen to be in accordance with encephalocele. It is rare for Dandy-Walker syndrome to occur together with occipital encephalocele. The authors present a case of Dandy-Walker syndrome together with occipital encephalocele. PMID- 18511900 TI - Unusual Gilbert's syndrome genotype in a Greek patient suffering from both Gilbert's syndrome and familial mediterranean fever. A case report. AB - Gilbert's syndrome is a genetically controlled non-hemolytic unconjugated hyperbilirubinemia, caused by reduced activity of UDP-glucoroniltransferase 1, an enzyme critical in bilirubin metabolism. Several molecular configurations may be implicated in a Gilbert's phenotype. Familial mediterranean fever (FMF) is an inherited acute relapsing inflammatory disorder, affecting Mediterranean and Middle East populations. The molecular basis of the disorder concerns the MEFV gene coding for a protein named pyrin; several point mutations of MEFV gene have been associated with the disease. The authors present an unusual patient co affected by both Gilbert's syndrome and FMF who carried a peculiar Gilbert's genotype. The coexistence of these two genetic conditions seems to be rare but interesting as the potentially overlapping clinical symptoms may rise interesting diagnostic problems. PMID- 18511901 TI - [Pedodontist and pediatrician in the management of the child affected by systemic pathologies]. PMID- 18511902 TI - [From the newborn infant to the child, to the adult: historia magistra vitae]. AB - The hypothesis of the prenatal programming of adult diseases took on increasing interest from the moment that, in the 60s, an epidemiological association was proposed between low birth weight and cardiovascular diseases. In the last 20 years it has been demonstrated that individuals with low weight, low stature and thinness at birth have a higher risk of developing cardiovascular diseases and type 2 diabetes. Animal and clinical studies are casting light on the biological mechanisms underlying the association between modified development in the uterus and diseases, and on how growth in adolescence and in adult life can modulate this initial proneness to disease. One of the mechanisms that has aroused most interest among researchers is the reduced number of nephrons, associated with low birth weight, which predisposes to glomerulosclerosis and increased systemic arterial pressure in adult life. A correlation has also been found between low weight at birth and peripheral resistance to insulin. Nevertheless, it is thought that modified prenatal development is only a predisposing factor, open to profound influences in the course of postnatal development. It has in fact been demonstrated that accelerated development in the period of infancy and adolescence can extend the initial condition of neonatal suffering. Indirectly these studies renew and at the same time extend the concept of prevention, a priority aim of the physician of the third millennium. PMID- 18511903 TI - Primary osteosarcoma of the scapula in an adolescent: an unusual presentation. PMID- 18511904 TI - Altered subcellular distribution of MSK1 induced by glucocorticoids contributes to NF-kappaB inhibition. AB - Glucocorticoids are widely used anti-inflammatory and immunomodulatory agents, of which the action mechanism is mainly based on interference of hormone-activated glucocorticoid receptor (GR) with the activity of transcription factors, such as nuclear factor-kappaB (NF-kappaB). In addition to the well described interaction based mutual repression mechanism between the GR and NF-kappaB, additional mechanisms are at play, which help to explain the efficacy of glucocorticoid mediated gene repression. In this respect, we found that glucocorticoids counteract the recruitment of activated Mitogen- and Stress-activated protein Kinase-1 (MSK1) at inflammatory gene promoters resulting in the inhibition of NF kappaB p65 transactivation and of concurrent histone H3 phosphorylation. Additionally, we observed that activated GR can trigger redistribution of nuclear MSK1 to the cytoplasm through a CRM1-dependent export mechanism, as a result of an interaction between liganded GR and activated MSK1. These findings unveil a novel aspect within the GR-mediated NF-kappaB-targeting anti-inflammatory mechanism. PMID- 18511905 TI - Sequential Cyk-4 binding to ECT2 and FIP3 regulates cleavage furrow ingression and abscission during cytokinesis. AB - Cytokinesis is a highly regulated and dynamic event that involves the reorganization of the cytoskeleton and membrane compartments. Recently, FIP3 has been implicated in targeting of recycling endosomes to the mid-body of dividing cells and is found required for abscission. Here, we demonstrate that the centralspindlin component Cyk-4 is a FIP3-binding protein. Furthermore, we show that FIP3 binds to Cyk-4 at late telophase and that centralspindlin may be required for FIP3 recruitment to the mid-body. We have mapped the FIP3-binding region on Cyk-4 and show that it overlaps with the ECT2-binding domain. Finally, we demonstrate that FIP3 and ECT2 form mutually exclusive complexes with Cyk-4 and that dissociation of ECT2 from the mid-body at late telophase may be required for the recruitment of FIP3 and recycling endosomes to the cleavage furrow. Thus, we propose that centralspindlin complex not only regulates acto-myosin ring contraction but also endocytic vesicle transport to the cleavage furrow and it does so through sequential interactions with ECT2 and FIP3. PMID- 18511906 TI - Break dosage, cell cycle stage and DNA replication influence DNA double strand break response. AB - DNA double strand breaks (DSBs) can be repaired by non-homologous end joining (NHEJ) or homology-directed repair (HR). HR requires nucleolytic degradation of 5' DNA ends to generate tracts of single-stranded DNA (ssDNA), which are also important for the activation of DNA damage checkpoints. Here we describe a quantitative analysis of DSB processing in the budding yeast Saccharomyces cerevisiae. We show that resection of an HO endonuclease-induced DSB is less extensive than previously estimated and provide evidence for significant instability of the 3' ssDNA tails. We show that both DSB resection and checkpoint activation are dose-dependent, especially during the G1 phase of the cell cycle. During G1, processing near the break is inhibited by competition with NHEJ, but extensive resection is regulated by an NHEJ-independent mechanism. DSB processing and checkpoint activation are more efficient in G2/M than in G1 phase, but are most efficient at breaks encountered by DNA replication forks during S phase. Our findings identify unexpected complexity of DSB processing and its regulation, and provide a framework for further mechanistic insights. PMID- 18511907 TI - Synergy of Eed and Tsix in the repression of Xist gene and X-chromosome inactivation. AB - X-chromosome inactivation (XCI) depends on the noncoding Xist gene. Xist transcription is negatively regulated by its antisense partner Tsix, whose disruption results in nonrandom XCI in females. However, males can maintain Xist in a repressed state without Tsix, indicating participation of additional factor(s) in the protection of the single male X from inactivation. Here, we provide evidence that the histone methyltransferase Eed is also involved in the process. Male embryonic stem cells with Eed-null and Tsix mutations (X(Delta)Y Eed-/-) showed Xist hyperactivation upon differentiation, whereas cells with either mutation alone did not. Impaired X-linked gene expression was observed in the X(Delta)Y Eed-/- ES cells at the onset of differentiation. The Xist promoter in the X(Delta)Y Eed-/- cells showed elevated histone H3-dimethyl lysine 4 modifications and lowered CpG methylation, which are characteristics of open chromatin. Hence, we identified Eed as an additional major player in the regulation of Xist expression. The synergy of Polycomb group proteins and antisense Tsix transcription in Xist gene regulation explains why males can repress Xist without Tsix. PMID- 18511908 TI - Identification of PCTA, a TGIF antagonist that promotes PML function in TGF-beta signalling. AB - The TGIF homoeodomain protein functions as an important negative regulator in the TGF-beta signalling pathway. The inhibitory function of TGIF is executed in part through its ability to sequester the tumour suppressor cytoplasmic promyelocytic leukaemia (cPML) in the nucleus, thereby preventing the phosphorylation of Smad2 by the activated TGF-beta type I receptor. Here, we report on the identification of PCTA (PML competitor for TGIF association), a TGIF antagonist that promotes TGF-beta-induced transcriptional and cytostatic responses. We provide evidence that PCTA functions in TGF-beta signalling by relieving the suppression of Smad2 phosphorylation by TGIF. Furthermore, we demonstrate that PCTA selectively competes with cPML for TGIF association, resulting in the accumulation of cPML in the cytoplasm, where it associates with SARA and coordinates the access of Smad2 for phosphorylation by the activated TGF-beta type I receptor. Thus, our findings on the mode of action of PCTA provide new and important insights into the molecular mechanism underlying the antagonistic interplay between TGIF and cPML in the TGF-beta signalling network. PMID- 18511909 TI - The interaction network of the chaperonin CCT. AB - The eukaryotic cytosolic chaperonin containing TCP-1 (CCT) has an important function in maintaining cellular homoeostasis by assisting the folding of many proteins, including the cytoskeletal components actin and tubulin. Yet the nature of the proteins and cellular pathways dependent on CCT function has not been established globally. Here, we use proteomic and genomic approaches to define CCT interaction networks involving 136 proteins/genes that include links to the nuclear pore complex, chromatin remodelling, and protein degradation. Our study also identifies a third eukaryotic cytoskeletal system connected with CCT: the septin ring complex, which is essential for cytokinesis. CCT interactions with septins are ATP dependent, and disrupting the function of the chaperonin in yeast leads to loss of CCT-septin interaction and aberrant septin ring assembly. Our results therefore provide a rich framework for understanding the function of CCT in several essential cellular processes, including epigenetics and cell division. PMID- 18511910 TI - Branch migration enzyme as a Brownian ratchet. AB - In recent years, it has been shown that helicases are able to perform functions beyond their traditional role in unwinding of double-stranded nucleic acids; yet the mechanistic aspects of these different activities are not clear. Our kinetic studies of Holliday junction branch migration catalysed by a ring-shaped helicase, T7 gp4, show that heterology of as little as a single base stalls catalysed branch migration. Using single-molecule analysis, one can locate the stall position to within a few base pairs of the heterology. Our data indicate that the presence of helicase alone promotes junction unfolding, which accelerates spontaneous branch migration, and individual time traces reveal complex trajectories consistent with random excursions of the branch point. Our results suggest that instead of actively unwinding base pairs as previously thought, the helicase exploits the spontaneous random walk of the junction and acts as a Brownian ratchet, which walks along duplex DNA while facilitating and biasing branch migration in a specific direction. PMID- 18511911 TI - Activated macrophages promote Wnt signalling through tumour necrosis factor-alpha in gastric tumour cells. AB - The activation of Wnt/beta-catenin signalling has an important function in gastrointestinal tumorigenesis. It has been suggested that the promotion of Wnt/beta-catenin activity beyond the threshold is important for carcinogenesis. We herein investigated the role of macrophages in the promotion of Wnt/beta catenin activity in gastric tumorigenesis. We found beta-catenin nuclear accumulation in macrophage-infiltrated dysplastic mucosa of the K19-Wnt1 mouse stomach. Moreover, macrophage depletion in Apc(Delta716) mice resulted in the suppression of intestinal tumorigenesis. These results suggested the role of macrophages in the activation of Wnt/beta-catenin signalling, which thus leads to tumour development. Importantly, the conditioned medium of activated macrophages promoted Wnt/beta-catenin signalling in gastric cancer cells, which was suppressed by the inhibition of tumour necrosis factor (TNF)-alpha. Furthermore, treatment with TNF-alpha induced glycogen synthase kinase 3beta (GSK3beta) phosphorylation, which resulted in the stabilization of beta-catenin. We also found that Helicobacter infection in the K19-Wnt1 mouse stomach caused mucosal macrophage infiltration and nuclear beta-catenin accumulation. These results suggest that macrophage-derived TNF-alpha promotes Wnt/beta-catenin signalling through inhibition of GSK3beta, which may contribute to tumour development in the gastric mucosa. PMID- 18511912 TI - Regulation of myosin light chain kinase expression by angiotensin II in hypertension. AB - BACKGROUND: Increased growth and contraction of vascular smooth muscle cells (VSMCs) are major abnormalities in many vascular disorders. To investigate the signaling pathways that mediate these processes, we studied the expression of smooth muscle myosin light chain kinase (smMLCK) in VSMCs. METHODS: Primary cultured VSMCs isolated from normotensive Wistar-Kyoto (WKY) rats were treated with angiotensin II (Ang II). smMLCK expression was examined in the cells using western blot analysis. In vivo, a specific inhibitor of smMLCK or MAP kinase kinase (MEK) was delivered to spontaneously hypertensive rats (SHRs) using an osmotic pump, and their blood pressures were measured using tail-cuff sphygmomanometry. RESULTS: Expression of smMLCK protein is rapidly increased by Ang II, an important agonist responsible for increased vasoconstriction and vascular remodeling, in concert with increased myosin light chain phosphorylation. Inhibiting Ang II type 1 (AT1) receptor, Ras, or MEK blocked the Ang II-induced increase in smMLCK expression. In vivo, inhibiting MEK decreased smMLCK expression, blood pressure, and vascular thickening in SHRs. Moreover, inhibiting smMLCK activity decreased blood pressure and smooth muscle mass in arteries in SHRs. CONCLUSIONS: The regulation of smMLCK expression by Ang II via Ras signaling is important in the regulation of vascular remodeling and blood pressure. Targeting this pathway could be an effective strategy for developing novel therapeutics to treat hypertension. PMID- 18511913 TI - Iodoacetamide-induced artifact mimics ubiquitination in mass spectrometry. PMID- 18511914 TI - Super-resolution for a 3D world. PMID- 18511915 TI - Turning fluorescent proteins into energy-saving light bulbs. PMID- 18511916 TI - Do-it-yourself guide: how to use the modern single-molecule toolkit. AB - Single-molecule microscopy has evolved into the ultimate-sensitivity toolkit to study systems from small molecules to living cells, with the prospect of revolutionizing the modern biosciences. Here we survey the current state of the art in single-molecule tools including fluorescence spectroscopy, tethered particle microscopy, optical and magnetic tweezers, and atomic force microscopy. We also provide guidelines for choosing the right approach from the available single-molecule toolkit for applications as diverse as structural biology, enzymology, nanotechnology and systems biology. PMID- 18511917 TI - Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy. AB - Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. Here we describe these techniques and illustrate them with examples highlighting current capabilities and limitations. PMID- 18511918 TI - A practical guide to single-molecule FRET. AB - Single-molecule fluorescence resonance energy transfer (smFRET) is one of the most general and adaptable single-molecule techniques. Despite the explosive growth in the application of smFRET to answer biological questions in the last decade, the technique has been practiced mostly by biophysicists. We provide a practical guide to using smFRET, focusing on the study of immobilized molecules that allow measurements of single-molecule reaction trajectories from 1 ms to many minutes. We discuss issues a biologist must consider to conduct successful smFRET experiments, including experimental design, sample preparation, single molecule detection and data analysis. We also describe how a smFRET-capable instrument can be built at a reasonable cost with off-the-shelf components and operated reliably using well-established protocols and freely available software. PMID- 18511919 TI - Laminar flow cells for single-molecule studies of DNA-protein interactions. AB - Microfluidic flow cells are used in single-molecule experiments, enabling measurements to be made with high spatial and temporal resolution. We discuss the fundamental processes affecting flow cell operation and describe the flow cells in use at present for studying the interaction of optically trapped or mechanically isolated, single DNA molecules with proteins. To assist the experimentalist in flow cell selection, we review the construction techniques and materials used to fabricate both single- and multiple-channel flow cells and the advantages of each design for different experiments. PMID- 18511920 TI - Nanotechnology in biology: big collaborations for a small world. PMID- 18511921 TI - Building the bridge from bench to bedside. PMID- 18511922 TI - Laying the regulatory path for advanced therapies. PMID- 18511925 TI - An audience with James Greenwood. PMID- 18511926 TI - Bendamustine. PMID- 18511928 TI - Drug development of MET inhibitors: targeting oncogene addiction and expedience. AB - The MET tyrosine kinase stimulates cell scattering, invasion, protection from apoptosis and angiogenesis, thereby acting as a powerful expedient for cancer dissemination. MET can also be genetically selected for the long-term maintenance of the primary transformed phenotype, and some tumours appear to be dependent on (or 'addicted' to) sustained MET activity for their growth and survival. Because of its dual role as an adjuvant, pro-metastatic gene for some tumour types and as a necessary oncogene for others, MET is a versatile candidate for targeted therapeutic intervention. Here we discuss recent progress in the development of molecules that inhibit MET function and consider their application in a subset of human tumours that are potentially responsive to MET-targeted therapies. PMID- 18511927 TI - Fatty acid-binding proteins: role in metabolic diseases and potential as drug targets. AB - Lipids are vital components of many biological processes and crucial in the pathogenesis of numerous common diseases, but the specific mechanisms coupling intracellular lipids to biological targets and signalling pathways are not well understood. This is particularly the case for cells burdened with high lipid storage, trafficking and signalling capacity such as adipocytes and macrophages. Here, we discuss the central role of lipid chaperones--the fatty acid-binding proteins (FABPs)--in lipid-mediated biological processes and systemic metabolic homeostasis through the regulation of diverse lipid signals, and highlight their therapeutic significance. Pharmacological agents that modify FABP function may provide tissue-specific or cell-type-specific control of lipid signalling pathways, inflammatory responses and metabolic regulation, potentially providing a new class of drugs for diseases such as obesity, diabetes and atherosclerosis. PMID- 18511929 TI - Emerging anti-insomnia drugs: tackling sleeplessness and the quality of wake time. AB - Sleep is essential for our physical and mental well being. However, when novel hypnotic drugs are developed, the focus tends to be on the marginal and statistically significant increase in minutes slept during the night instead of the effects on the quality of wakefulness. Recent research on the mechanisms underlying sleep and the control of the sleep-wake cycle has the potential to aid the development of novel hypnotic drugs; however, this potential has not yet been realized. Here, we review the current understanding of how hypnotic drugs act, and discuss how new, more effective drugs and treatment strategies for insomnia might be achieved by taking into consideration the daytime consequences of disrupted sleep. PMID- 18511930 TI - Why prevent invasive Candida infections? PMID- 18511931 TI - VATS management of an enlarging multicystic pneumatocele. AB - A multicystic pneumatocele progressively enlarged when the patient required positive pressure ventilation for an intercurrent respiratory syncytial virus infection. Video Assisted Thoracoscopic Surgery was used to visualize the pneumatocele for cannulation. One chamber of the pneumatocele was cannulated with a pigtail catheter and another large chamber ruptured, without cannulation. The multicystic pneumatocele resolved with this therapy. PMID- 18511932 TI - Congenital adenohypophysis aplasia. PMID- 18511933 TI - Hif-2alpha mediates UV-induced apoptosis through a novel ATF3-dependent death pathway. AB - In this study, we describe a novel activating transcription factor 3 (ATF3) dependent death pathway triggered by ultraviolet (UV) irradiation. We demonstrate that ATF3 contributes to UV-induced apoptosis through the regulation of hypoxia inducible factor (Hif)-2alpha expression, which in turn induces the expression of proapoptotic genes, such as Caspase7 or TRAIL (tumor necrosis factor (ligand) superfamily, member 10). Gain of function of Hif-2alpha as well as ATF3 is sufficient to trigger cell death, whereas loss of function of both proteins drastically inhibits UV-induced apoptosis. Repression of Hif-2alpha strongly impairs ATF3-mediated death, providing evidences that Hif-2alpha is the major death effector of ATF3. In addition, Hif-1alpha, already known as a proapoptotic gene, upon UV irradiation, is not able to compensate for the lack of Hif-2alpha expression, thereby confirming the major contribution of Hif-2alpha in UV mediated cell death. We further demonstrate that this cascade of gene activation depends on p38 and c-Jun N-terminal kinase (JNK) activity. Impairment of such a pathway is likely to contribute to oncogenesis by promoting survival of cells that could accumulate severe chromosomal alterations. PMID- 18511934 TI - HCCS1 overexpression induces apoptosis via cathepsin D and intracellular calcium, and HCCS1 disruption in mice causes placental abnormality. AB - Hepatocellular carcinoma suppressor 1 (HCCS1) was discovered as a novel tumor suppressor gene. We recently observed that adenovirus-mediated gene transfer of HCCS1 leads to cytotoxicity to human hepatocarcinoma cells. Here, we have demonstrated that adenovirus-mediated overexpression of HCCS1 induces apoptosis in hepatocarcinoma cells and have further characterized the apoptotic cascade. The results showed that lysosomal cathepsin D is released into the cytosol in response to HCCS1 overexpression and consequently triggers Bax insertion into the mitochondrial membrane, which leads to the release of cytochrome c. In addition, HCCS1 overexpression can induce an increase in intracellular free Ca(2+) concentration, which also results in cytochrome c release. The released cytochrome c activates downstream caspases, leading to the occurrence of the late stages of apoptosis. Moreover, we demonstrated that the disruption of HCCS1 in mice leads to embryonic lethality, accompanied by abnormal labyrinth architecture resulting from the excessive proliferation of trophoblast cells in the placenta. These results suggest that HCCS1 plays a role in apoptosis regulation and development. PMID- 18511935 TI - Defective MHC class I antigen surface expression promotes cellular survival through elevated ER stress and modulation of p53 function. AB - Defects in Major Histocompatibility class I cell surface expression is thought to allow escape of tumor cells from immune surveillance. Hitherto, it is unclear whether this deficiency confers immune-independent survival advantage. We show here that class I cell surface expression deficiency due to defects in beta2 microglobulin or the transporter-associated with antigen processing (TAP) results in resistance to apoptosis in response to various cytotoxic signals. Reduced apoptosis correlated with altered p53 activation, which was due to compromised nuclear translocation of p53. Binding of p53 to glycogen synthase kinase-3beta (GSK3beta), which is known to phosphorylate and lead to cytoplasmic sequestration of p53, was enhanced in these cells. Consistently, endoplasmic reticulum (ER) stress, which promotes binding of p53 to GSK3beta was constitutively elevated in the absence of class I cell surface expression. Taken together, the results suggest a non-immunological causal role for defective class I cell surface expression in regulating cellular survival in a p53-dependent manner, through the upregulation of ER stress, which could be another mechanism leading to carcinogenesis. PMID- 18511936 TI - Targeting the oncogene and kinome chaperone CDC37. AB - CDC37 is a molecular chaperone that physically stabilizes the catalytic domains found in protein kinases and is therefore a wide-spectrum regulator of protein phosphorylation. It is also an overexpressed oncoprotein that mediates carcinogenesis by stabilizing the compromised structures of mutant and/or overexpressed oncogenic kinases. Recent work shows that such dependency of malignant cells on increased CDC37 expression is a vulnerability that can be targeted in cancer by agents that deplete or inhibit CDC37. CDC37 is thus a candidate for broad-spectrum molecular cancer therapy. PMID- 18511937 TI - Exploring the role of cancer stem cells in radioresistance. AB - Radiobiological research over the past decades has provided evidence that cancer stem cell content and the intrinsic radiosensitivity of cancer stem cells varies between tumours, thereby affecting their radiocurability. Translation of this knowledge into predictive tests for the clinic has so far been hampered by the lack of methods to discriminate between stem cells and non-stem cells. New technologies allow isolation of cells expressing specific surface markers that are differentially expressed in tumour cell subpopulations that are enriched for cancer stem cells. Combining these techniques with functional radiobiological assays holds the potential to elucidate the role of cancer stem cells in radioresistance in individual tumours, and to use this knowledge for the development of predictive markers for optimization of radiotherapy. PMID- 18511938 TI - In situ collagen assembly for integrating microfabricated three-dimensional cell seeded matrices. AB - Microscale fabrication of three-dimensional (3D) extracellular matrices (ECMs) can be used to mimic the often inhomogeneous and anisotropic properties of native tissues and to construct in vitro cellular microenvironments. Cellular contraction of fibrous natural ECMs (such as fibrin and collagen I) can detach matrices from their surroundings and destroy intended geometry. Here, we demonstrate in situ collagen fibre assembly (the nucleation and growth of new collagen fibres from preformed collagen fibres at an interface) to anchor together multiple phases of cell-seeded 3D hydrogel-based matrices against cellular contractile forces. We apply this technique to stably interface multiple microfabricated 3D natural matrices (containing collagen I, Matrigel, fibrin or alginate); each phase can be seeded with cells and designed to permit cell spreading. With collagen-fibre-mediated interfacing, microfabricated 3D matrices maintain stable interfaces (the individual phases do not separate from each other) over long-term culture (at least 3 weeks) and support spatially restricted development of multicellular structures within designed patterns. The technique enables construction of well-defined and stable patterns of a variety of 3D ECMs formed by diverse mechanisms (including temperature-, ion- and enzyme-mediated crosslinking), and presents a simple approach to interface multiple 3D matrices for biological studies and tissue engineering. PMID- 18511939 TI - Feast or famine: the global regulator DasR links nutrient stress to antibiotic production by Streptomyces. AB - Members of the soil-dwelling prokaryotic genus Streptomyces produce many secondary metabolites, including antibiotics and anti-tumour agents. Their formation is coupled with the onset of development, which is triggered by the nutrient status of the habitat. We propose the first complete signalling cascade from nutrient sensing to development and antibiotic biosynthesis. We show that a high concentration of N-acetylglucosamine-perhaps mimicking the accumulation of N acetylglucosamine after autolytic degradation of the vegetative mycelium-is a major checkpoint for the onset of secondary metabolism. The response is transmitted to antibiotic pathway-specific activators through the pleiotropic transcriptional repressor DasR, the regulon of which also includes all N acetylglucosamine-related catabolic genes. The results allowed us to devise a new strategy for activating pathways for secondary metabolite biosynthesis. Such 'cryptic' pathways are abundant in actinomycete genomes, thereby offering new prospects in the fight against multiple drug-resistant pathogens and cancers. PMID- 18511940 TI - Crucial structural role for the PH and C1 domains of the Vav1 exchange factor. AB - The Vav family of proteins are guanine nucleotide exchange factors (GEFs) for the Rho family of GTPases, which regulate various cellular functions, including T cell activation. They contain a catalytic Dbl homology (DH) domain that is invariably followed by a pleckstrin homology (PH) domain, which is often required for catalytic activity. Vav proteins are the first GEFs for which an additional C1 domain is required for full biological activity. Here, we present the structure of a Vav1 fragment comprising the DH-PH-C1 domains bound to Rac1. This structure shows that the PH and C1 domains form a single structural unit that packs against the carboxy-terminal helix of the DH domain to stabilize its conformation and to promote nucleotide exchange. In contrast to previous reports, this structure shows that there are no direct contacts between the GTPase and C1 domain but instead suggests new mechanisms for the regulation of Vav1 activity. PMID- 18511941 TI - Technology assessment in radiation oncology: time for reassessment? PMID- 18511942 TI - EGCG redirects amyloidogenic polypeptides into unstructured, off-pathway oligomers. AB - The accumulation of beta-sheet-rich amyloid fibrils or aggregates is a complex, multistep process that is associated with cellular toxicity in a number of human protein misfolding disorders, including Parkinson's and Alzheimer's diseases. It involves the formation of various transient and intransient, on- and off-pathway aggregate species, whose structure, size and cellular toxicity are largely unclear. Here we demonstrate redirection of amyloid fibril formation through the action of a small molecule, resulting in off-pathway, highly stable oligomers. The polyphenol (-)-epigallocatechin gallate efficiently inhibits the fibrillogenesis of both alpha-synuclein and amyloid-beta by directly binding to the natively unfolded polypeptides and preventing their conversion into toxic, on pathway aggregation intermediates. Instead of beta-sheet-rich amyloid, the formation of unstructured, nontoxic alpha-synuclein and amyloid-beta oligomers of a new type is promoted, suggesting a generic effect on aggregation pathways in neurodegenerative diseases. PMID- 18511943 TI - Nonprocessive methylation by Dot1 leads to functional redundancy of histone H3K79 methylation states. AB - Whereas mono-, di- and trimethylation states of lysines on histones typically have specific functions, no specific functions have been attributed so far to the different methylation states of histone H3 Lysine 79 (H3K79) generated by Dot1. Here we show that Dot1, in contrast to other known histone methyltransferases, introduces multiple methyl groups via a nonprocessive mechanism. The kinetic mechanism implies that the H3K79 methylation states cannot be generated independently, suggesting functional redundancy. Indeed, gene silencing in yeast, which is dependent on Dot1, relied on global H3K79 methylation levels and not on one specific methylation state. Furthermore, our findings suggest that histone H2B ubiquitination affects H3K79 trimethylation by enhancing synthesis of all H3K79 methylation states. Our results suggest that multiple methylation of H3K79 leads to a binary code, which is expected to limit the possibilities for regulation by putative demethylases or binding proteins. PMID- 18511944 TI - Long single alpha-helical tail domains bridge the gap between structure and function of myosin VI. AB - Myosin VI has challenged the lever arm hypothesis of myosin movement because of its ability to take approximately 36-nm steps along actin with a canonical lever arm that seems to be too short to allow such large steps. Here we demonstrate that the large step of dimeric myosin VI is primarily made possible by a medial tail in each monomer that forms a rare single alpha-helix of approximately 10 nm, which is anchored to the calmodulin-bound IQ domain by a globular proximal tail. With the medial tail contributing to the approximately 36-nm step, rather than dimerizing as previously proposed, we show that the cargo binding domain is the dimerization interface. Furthermore, the cargo binding domain seems to be folded back in the presence of the catalytic head, constituting a potential regulatory mechanism that inhibits dimerization. PMID- 18511945 TI - The central unit within the 19S regulatory particle of the proteasome. AB - The 26S proteasome is a multisubunit enzyme composed of a cylindrical catalytic core (20S) and a regulatory particle (19S) that together perform the essential degradation of cellular proteins tagged by ubiquitin. To date, however, substrate trajectory within the complex remains elusive. Here we describe a previously unknown functional unit within the 19S, comprising two subunits, Rpn1 and Rpn2. These toroids physically link the site of substrate recruitment with the site of proteolysis. Rpn2 interfaces with the 20S, whereas Rpn1 sits atop Rpn2, serving as a docking site for a substrate-recruitment factor. The 19S ATPases encircle the Rpn1-Rpn2 stack, covering the remainder of the 20S surface. Both Rpn1-Rpn2 and the ATPases are required for substrate translocation and gating of the proteolytic channel. Similar pairing of units is found in unfoldases and nuclear transporters, exposing common features of these protein nanomachines. PMID- 18511946 TI - Age-related macular degeneration is associated with an unstable ARMS2 (LOC387715) mRNA. AB - Age-related macular degeneration (AMD) is a prevalent multifactorial disorder of the central retina. Genetic variants at two chromosomal loci, 1q31 and 10q26, confer major disease risks, together accounting for more than 50% of AMD pathology. Signals at 10q26 center over two nearby genes, ARMS2 (age-related maculopathy susceptibility 2, also known as LOC387715) and HTRA1 (high temperature requirement factor A1), suggesting two equally probable candidates. Here we show that a deletion-insertion polymorphism in ARMS2 (NM_001099667.1:c.(*)372_815del443ins54) is strongly associated with AMD, directly affecting the transcript by removing the polyadenylation signal and inserting a 54-bp element known to mediate rapid mRNA turnover. As a consequence, expression of ARMS2 in homozygous carriers of the indel variant is not detectable. Confirming previous findings, we demonstrate a mitochondrial association of the normal protein and further define its retinal localization to the ellipsoid region of the photoreceptors. Our data suggest that ARMS2 has a key role in AMD, possibly through mitochondria-related pathways. PMID- 18511947 TI - Strong association of de novo copy number mutations with sporadic schizophrenia. AB - Schizophrenia is an etiologically heterogeneous psychiatric disease, which exists in familial and nonfamilial (sporadic) forms. Here, we examine the possibility that rare de novo copy number (CN) mutations with relatively high penetrance contribute to the genetic component of schizophrenia. We carried out a whole genome scan and implemented a number of steps for finding and confirming CN mutations. Confirmed de novo mutations were significantly associated with schizophrenia (P = 0.00078) and were collectively approximately 8 times more frequent in sporadic (but not familial) cases with schizophrenia than in unaffected controls. In comparison, rare inherited CN mutations were only modestly enriched in sporadic cases. Our results suggest that rare de novo germline mutations contribute to schizophrenia vulnerability in sporadic cases and that rare genetic lesions at many different loci can account, at least in part, for the genetic heterogeneity of this disease. PMID- 18511948 TI - NAD(P)H:quinone oxidoreductase 1 NQO1*2 genotype (P187S) is a strong prognostic and predictive factor in breast cancer. AB - NQO1 guards against oxidative stress and carcinogenesis and stabilizes p53. We find that a homozygous common missense variant (NQO1(*)2, rs1800566(T), NM_000903.2:c.558C>T) that disables NQO1 strongly predicts poor survival among two independent series of women with breast cancer (P = 0.002, N = 1,005; P = 0.005, N = 1,162), an effect particularly evident after anthracycline-based adjuvant chemotherapy with epirubicin (P = 7.52 x 10(-6)) and in p53-aberrant tumors (P = 6.15 x 10(-5)). Survival after metastasis was reduced among NQO1(*)2 homozygotes, further implicating NQO1 deficiency in cancer progression and treatment resistance. Consistently, response to epirubicin was impaired in NQO1(*)2-homozygous breast carcinoma cells in vitro, reflecting both p53-linked and p53-independent roles of NQO1. We propose a model of defective anthracycline response in NQO1-deficient breast tumors, along with increased genomic instability promoted by elevated reactive oxygen species (ROS), and suggest that the NQO1 genotype is a prognostic and predictive marker for breast cancer. PMID- 18511949 TI - The effects of chronic corticosterone on hippocampal astrocyte numbers: a comparison of male and female Wistar rats. AB - Chronic stress an/or glucocorticoid administration produces atrophy of hippocampal neurons. However, evidence of the impact of glucocorticoids on glial cells, especially in both males and females, is limited. In the present study, we investigated the total percentage body weight, hippocampal volume and hippocampal astrocyte numbers following chronic corticosterone treatment in male and female Wistar rats. Males had greater left and right hippocampal volumes overall, but no effect on hippocampal volume was seen after corticosterone treatment. Total body weight was dose-dependently lower in both sexes, but the decrease was more prominent in male rats. Corticosterone treatment dose-dependently increased astrocyte numbers in the CA1 region, but not in the lateral and medial CA3 hippocampal regions. This increase was similar in both male and female rats. The astrogliosis observed following chronic corticosterone may have implications for extrasynaptic communication and neuron-glia interactions and is similar to changes in the astrocytic population observed in aged rats. PMID- 18511950 TI - Cell death and neuronal arborization upon quercetin treatment in rat neurons. AB - Quercetin, one of the major flavonoids, exhibits many beneficial effects on human organism as antihistamine, antioxidant, anti-inflammatory, anticancer and antiviral drug. It is recommended as supplement of healthy diet but still the knowledge of its beneficial effect on normal cells is not satisfactory. We decided to examine the effect of flavonoid on neurons morphology and their susceptibility to cell death. Fractal analysis of rat neurons revealed that 24 hours long incubation with quercetin diminished neuronal arborisation in cortical neurons. Neurons also appeared to be very sensitive to cell death after flavonoid treatment in concentration dependent manner. Over 50% of cells died after incubation with 15 mu/ml of flavonoid while 1 mu/ ml of quercetin induced cell death only in 5%. Staining with Hoechst 33342 and propidium ioidide revealed the two types of cell death: apoptosis and necrosis. The number of apoptotic cells was comparable with necrotic ones. These results suggest toxic effect of quercetin on neurons what should be taken into consideration in further studies on using quercetin as therapeutic agent. PMID- 18511951 TI - Expression of Angiopoietin-1 and the receptor Tie-2 mRNA in rat brains following intracerebral hemorrhage. AB - Angiopoietin-1 (Ang-1) belongs to a novel family of endothelial growth factors that function as ligands for an endothelialspecific receptor tyrosine kinase (Tie 2). The Ang-/Tie-2 system may contribute to angiogenesis and vascular remodeling by mediating interactions of endothelial cells with smooth muscle cells and pericytes. The spatial distribution and temporal expression of Ang-1 and Tie-2 in the rat brain were studied following collagenase-induced intracerebral hemorrhage (ICH) by immunohistochemistry and reverse transcription-polymerase chain (RT-PCR) analysis, respectively. Immunohistochemical analysis revealed that some Ang-1 or Tie-2-positive dilated vessels resided around the hematoma and extended into the clot. RT-PCR analysis showed that Ang-1 and Tie-2 mRNA signal was detected at 2 days and persisted for 28 days after ICH. These findings suggest that ICH could lead to upregulation of Ang-1 and the receptor Tie-2 mRNA. PMID- 18511952 TI - Spatiotemporal dynamics of astroglial and microglial responses after photothrombotic stroke in the rat brain. AB - The effects of photothrombotic stroke in primary somatosensory cortex on astroglial and microglial activation in various regions of lesioned brain were examined at different time points, using immunohistochemistry and lectin binding. The increase in GFAP expression was observed exclusively in the ipsilateral hemisphere, both in the perilesional area and cortical regions distant from the infarct. This remote increase was detectable up to sixty days after the infarct. Transient GFAP elevation was also found in the hippocampus one day after photothrombosis, whereas it was more prolonged in amygdala, as demonstrated at four days after lesion. In contrast to a widespread astrocytic activation, the microglial response was shortlasting and local, confined to lesion and perilesional area. Widespread and prolonged activation of astrocytes after stroke may provide factors promoting slowly developing recovery processes in the whole brain, while microglial response seems to be involved in local repair and removal of cellular debris. PMID- 18511953 TI - Preconditioning reduces hypoxia-evoked alterations in glutamatergic Ca2+ signaling in rat cortex. AB - The aims of this study were (1) to characterize calcium signaling in rat cortex induced by repeated in vitro application of the glutamatergic agonists L glutamate, NMDA, AMPA and DHPG, (2) to analyze the influence of transient severe hypobaric hypoxia (180 Torr) administered in vivo on calcium responses to stimulation of glutamate receptors by their agonists, and (3) to evaluate the effects of preconditioning with intermittent mild hypobaric hypoxia (360 Torr) 24 h before the severe hypoxia, on these Ca2+ responses. Intracellular Ca2+ dynamics was studied using the fluorescent probes fura-2 and chlortetracycline to monitor free and bound calcium (Cai and Cab) respectively. In control cortical slices, application of L-glutamate, NMDA and AMPA induced concomitant increases in Cai and Cab, reflecting Ca2+ influx and its intracellular accumulation in neurons. DHPG, an agonist of group I mGlu receptors induced a decrease in Cab accompanied by a rise in Cai levels, indicating Ca2+ mobilization. In cortical slices collected 24 h after severe hypoxia, the responses of Cab to glutamate administration were increased, DHPG-induced shifts were reversed, the increase in Cab after the first application of AMPA was reduced, while after the second, Cab rises were potentiated, and the increases in Cab evoked by NMDA application were slightly suppressed. The alterations of responses in Cab to the selective agonists were completely prevented by preconditioning with mild hypoxia. Our results suggest that protection of normal glutamatergic calcium signaling contributes to tolerance to hypoxia induced by preconditioning. PMID- 18511954 TI - Subjective mood estimation co-varies with spectral power EEG characteristics. AB - Co-variation between subjectively estimated mood/activation and EEG characteristics, based on spectral power parameters, was investigated. Subjective estimation of mood was made by using Thayer's Activation-Deactivation Adjective Checklist, which yielded two dimensions: Energy-Tiredness (with Energy pole having positive valence connotation) and Tension-Calmness (negative connotation for Tension). A within-subject design with two sessions of EEG recording immediately followed by mood assessment was applied. These were separated by a cognitive task, introduced in order to modify the subjects' mood. The correlations between changes in mood estimation and changes in EEG spectral power parameters were calculated. Both ADACL dimensions co-varied with EEG in a specific way according to frequency and localization. Subjective estimation of Energy correlated negatively with alpha1 and, surprisingly, positively with delta, theta1 as well as theta2 relative power. Estimation of Tension correlated positively with theta1 and beta1, and negatively with alpha2 relative power. Presented results suggest that the adjective description of mood has objectively measurable brain correlates in the EEG. PMID- 18511955 TI - Emotion-dependent modulation of interference processes: an fMRI study. AB - We studied the effects of experimentally induced emotions on inhibitory control using functional magnetic resonance tomography (fMRI). The Simon task used involves two conditions with different attentional demands and is a well established paradigm for studying inhibitory control. Incompatible trials demand cognitive control for resolving interference. Compatible trials need no inhibitory control. Twelve participants viewed a series of affective pictures inducing positive, negative or neutral affects. Between the picture blocks, participants performed either incompatible or compatible trials. Behavioral and fMRI data revealed an impact of negative emotions only on the processing of ncompatible trials. Subjects made more errors and showed less activation of brain areas associated with task performance. There was no effect of positive emotions neither on compatible nor incompatible trials. The results first showed that especially the processing of negative emotions is resource competing and secondly that the competition concerns only the controlled route of cognitive processing. PMID- 18511956 TI - Auditory language comprehension of temporally reversed speech signals in native and non-native speakers. AB - Neuropsychological studies in brain-injured patients with aphasia and children with specific language-learning deficits have shown the dependence of language comprehension on auditory processing abilities, i.e. the detection of temporal order. An impairment of temporal-order perception can be simulated by time reversing segments of the speech signal. In our study, we investigated how different lengths of time-reversed segments in speech influenced comprehension in ten native German speakers and ten participants who had acquired German as a second language. Results show that native speakers were still able to understand the distorted speech at segment lengths of 50 ms, whereas non-native speakers only could identify sentences with reversed intervals of 32 ms duration. These differences in performance can be interpreted by different levels of semantic and lexical proficiency. Our method of temporally-distorted speech offers a new approach to assess language skills that indirectly taps into lexical and semantic competence of non-native speakers. PMID- 18511957 TI - Emotion perception and interpersonal behavior in epilepsy patients after unilateral amygdalohippocampectomy. AB - Aim of this study was to examine the relation between perception of facial expressions and interpersonal behavior in epilepsy patients after unilateral amygdalohippocampectomy (AH). Nine patients with unilateral amygdalohippocampectomy and 14 controls completed a forced-choice emotional recognition task, in which morphed facial emotional expressions were shown at different emotional intensities, and a self-report questionnaire of interpersonal behavior. Face perception and depressive symptoms were also taken into account. Compared to normal controls, patients were less sensitive in the recognition of fearful and disgusted facial expressions, in line with previous reports. These impairments were only minimally correlated with self-report interpersonal behavior. In all, unilateral damage to the amygdala and medial temporal lobe results in subtle emotion recognition impairments, but these deficits do not appear to extend to self-reported impairments in everyday interpersonal behavior. Further studies need to explore in more detail the effects of these subtle recognition problems on daily social intercourse. PMID- 18511958 TI - A putative role of the Amyloid Precursor Protein Intracellular Domain (AICD) in transcription. AB - Amyloid Precursor Protein (APP) Intracellular Domain (AICD) is the product of APP processing realized by alpha- or beta-secretases and gamma-secretase. It was shown that AICD is able to interact with several proteins which regulate its stability and cellular localization. The Fe65 adaptor protein translocates AICD into nucleus where the APP-Fe65-Tip60 ternary complex may activate transcription of target genes. In the light of recent studies AICD seems to be another product of APP proteolysis endowed with important biological functions that may contribute to Alzheimer's disease pathology. PMID- 18511959 TI - Facing facts: neuronal mechanisms of face perception. AB - The face is one of the most important stimuli carrying social meaning. Thanks to the fast analysis of faces, we are able to judge physical attractiveness and features of their owners' personality, intentions, and mood. From one's facial expression we can gain information about danger present in the environment. It is obvious that the ability to process efficiently one's face is crucial for survival. Therefore, it seems natural that in the human brain there exist structures specialized for face processing. In this article, we present recent findings from studies on the neuronal mechanisms of face perception and recognition in the light of current theoretical models. Results from brain imaging (fMRI, PET) and electrophysiology (ERP, MEG) show that in face perception particular regions (i.e. FFA, STS, IOA, AMTG, prefrontal and orbitofrontal cortex) are involved. These results are confirmed by behavioral data and clinical observations as well as by animal studies. The developmental findings reviewed in this article lead us to suppose that the ability to analyze face-like stimuli is hard-wired and improves during development. Still, experience with faces is not sufficient for an individual to become an expert in face perception. This thesis is supported by the investigation of individuals with developmental disabilities, especially with autistic spectrum disorders (ASD). PMID- 18511960 TI - Electrophysiological description of mechanisms determining synaptic transmission and its modulation. AB - Signal integration in neurons is a complex process that depends on e.g. the kinetics of synaptic currents, distribution of synaptic connections as well as passive and excitatory membrane properties. The time course of synaptic currents is largely determined by the kinetics of the postsynaptic receptors and the time course of synaptic neurotransmitter concentration. The analysis of current responses to rapid agonist applications provides the means to study the ligand gated receptor gating but experimentally based estimation of neurotransmitter transient at central synapses was an important challenge during the last decade. Both theoretical as well as experimentally based approaches indicated that synaptic agonist transient is very brief, implying that the activation of postsynaptic receptors occurs in conditions of extreme non-equilibrium. Such a dynamic pattern of activation of postsynaptic receptors has a crucial impact not only on the kinetics of synaptic currents but also on their susceptibility to pharmacological modulation. PMID- 18511961 TI - Molecular basis of dendritic arborization. AB - The pattern of dendritic branching along with the receptor and channel composition and density of synapses regulate the electrical properties of neurons. Abnormalities in dendritic tree development lead to serious dysfunction of neuronal circuits and, consequently, the whole nervous system. Not surprisingly, the complicated and multi-step process of dendritic arbor development is highly regulated and controlled at every stage by both extrinsic signals and intrinsic molecular mechanisms. In this review, we analyze the molecular mechanisms that contribute to cellular processes that are crucial for the proper formation and stability of dendritic arbors, in such distant organisms as insects (e.g. Drosophila melanogaster) amphibians (Xenopus laevis) and mammals. PMID- 18511962 TI - Molecular mechanisms of dendritic spine development and maintenance. AB - The large majority of excitatory synapses are located on dendritic spines which are discrete membrane protrusions present on neuronal dendrites. Interestingly the highly heterogeneous morphology of dendritic spines is thought to be the morphological basis for synaptic plasticity associated to learning and memory formation. Indeed dendritic spines structure is regulated by molecular mechanisms that are fine tuned and adjusted according to level and direction of synaptic activity, development, specific brain region, and different experimental behavioral conditions. This supports the idea that reciprocal changes between the structure and function of spines impact both local and global integration of signals within dendrites. An increasing number of proteins have been found to be morphogens for dendritic spines and provided new insights into the molecular mechanisms regulating spine formation and morphology. Thus determining the mechanisms that regulate spine formation and morphology is essential for understanding the cellular changes that underlie learning and memory in normal and pathological conditions. PMID- 18511963 TI - Dendrites as separate compartment - local protein synthesis. AB - The article summarizes the most meaningful studies which have provided evidence that protein synthesis in neurons can occur not only in cell perikarya but also locally in dendrites. The presence of the complete machinery required to synthesize cytoplasmic and integral membrane proteins in dendrites, identification of binding proteins known to mediate mRNA trafficking in dendrites and the ability to trigger "on-site" translation make it possible for the synthesis of particular proteins to be regulated by synaptic signals. Until now over 100 different mRNAs coding the proteins involved in neurotransmission and modulation of synaptic activity have been identified in dendrites. Local protein synthesis is postulated to provide the basic mechanism of fast changes in the strength of neuronal connections and to be an important factor in the molecular background of synaptic plasticity, giving rise to enduring changes in synaptic function, which in turn play a role in local homeostatic responses. Local protein synthesis points to some autonomy of dendrites which makes them "the brains of the neurons" (Jim Eberwine; from the interview with J. Eberwine - Barinaga 2000). PMID- 18511964 TI - External and internal inputs affecting plasticity of dendrites and axons of the fly's neurons. AB - Neurons and glial cells in the fly's visual system exhibit circadian rhythms through changes in shape and size. Moreover, the number of synaptic contacts between these cells changes during the day and night and in the case of one type of synapses, feedback synapses, is maintained under constant conditions indicating an endogenous origin of this rhythm. The structural changes described above, involving the oscillations in the number of synapses and the size of interneurons and glial cells, are examples of plasticity in the central nervous system driven by internal inputs from a circadian clock and by external stimuli such as light. They are also modulated by visual and other sensory stimuli and by motor activity. PMID- 18511965 TI - Injury induced dendritic plasticity in the mature central nervous system. AB - Injury to the mature central nervous system (CNS) induces a series of transient changes leading not only to death of neurons, but also to spontaneous rearrangement of the affected network. One of such pro plastic events, detected following injury, is an increased level of neurotrophins. Neurotrophins are a family of proteins involved in survival and outgrowth processes. The other one, more difficult to observe, is a change in the complexity of the dendritic tree, causing arborization or pruning, depending on many circumstances: i.e. lesion etiology. Subsequent therapies like enriched environment or locomotor exercise bring about a functional improvement, which was found to further increase the neurotrophin level and induced additional arborization of dendrites. Another important consequence of damage to CNS connections is deafferentation, shown to induce a down regulation of outgrowth inhibitors. Their suppression in turn may facilitate dendritic plasticity. Taken together, these factors may contribute to enhanced plasticity in the injured mature CNS. Thus the proper use of endogenously increased plastic potential seems to be important for design and optimizing therapeutic strategies. Further investigation of mechanisms involved in switching on plasticity may help to improve on existing therapies and find new ways to obtain better recovery following injury. PMID- 18511966 TI - [The National Theatre]. PMID- 18511967 TI - [Prostatic cancer--who needs treatment?]. PMID- 18511968 TI - [Variable prescription of opioids to cancer patients in Norway]. AB - BACKGROUND: We examined prescription of the opioids used most often in cancer pain relief in the Norwegian counties, doctors' use of the law section concerning reimbursement of medicine to incurable far advanced cancer patients, and the prescription of analgesics with codein to this patient group. MATERIAL AND METHOD: Data were retrieved from The Norwegian prescription database for the period 01.01.04 to 31.08.05 (NorPD) on every prescription of slow and instant release morphine and oxycodone for oral administration and of transdermal fentanyl patches. SPSS was used for analyses. RESULTS: 5,675,038 Defined Daily Doses (DDD) of strong opioids were prescribed to 20,797 patients (46% of these to cancer patients) from 01.01.04 to 31.08.05. The county that prescribed most had prescribed 2.83 times more opioids than the county that had prescribed the least. Use of the law section concerning prescription of opioids indicates that these are prescribed as much for non-malignant diseases as for cancer. "Codein analgesics" are also commonly prescribed to cancer patients. INTERPRETATION: Prescription of strong opioids varies substantially in Norway's counties. Doctors' knowledge, attitude, geographical and cultural differences may be some reasons for this. Knowledge about and use of the law section concerning reimbursement is probably not good enough. Increased awareness of how to treat cancer pain is important go give a good treatment offer to all in Norway. PMID- 18511969 TI - [High dose rate brachytherapy in prostate cancer in Norway]. AB - BACKGROUND: A correlation exists between applied dose to the prostate and local tumour control. External radiotherapy of the prostate implies administering curative doses near the upper limit of normal tissue tolerance. Brachytherapy with a high dose rate permits an escalation of dose within the prostate without increasing the risk of side effects to the surrounding rectum and bladder. This article presents a study of the first 100 patients in Norway with localized/locally advanced prostate cancer treated with high dose-rate brachytherapy combined with external radiotherapy. MATERIAL AND METHODS: Patients belonging to an intermediate or high risk group and patients in whom radiotherapy was expected to give rise to increased toxicity (irrespective of the clinical stage) were included. Several hollow steel needles were implanted through the perineum into the gland during general anaesthesia. A small Iridium source with a short irradiation length was introduced stepwise and temporarily into each steel needle, according to a meticulate dosing plan. All patients came to an outpatient control 3-5 months after the combined treatment. RESULTS AND INTERPRETATION: Median follow-up was 8 months. Acute side effects were scarce and few complications from the rectum were seen. The observation time was too short to evaluate the relapse-free survival. High dose rate brachytherapy is indicated in patients with prostate cancer of an intermediate or high risk or if a radiation dose with a full external beam proposes a hazard to the patient. Combined radiotherapy (external beam and high dose rate brachytherapy) is considered to be a standard treatment at the Norwegian Radium Hospital. PMID- 18511970 TI - [Epithelial-to-mesenchymal transition and prostate cancer]. AB - BACKGROUND: Prostate cancer is highly prevalent and is a frequent cause of death in men. As for most other cancers the prognosis is largely determined by the occurrence of metastases. Future treatment of prostate cancer should focus on inhibition of the cancer cells' ability to invade surrounding tissues--and to metastasise. In order to develop such therapies, it is important to unveil the mechanisms that lead to an invasive phenotype. Development of invasive tumours resemble processes involved in embryonic development, e.g. during formation of the mesoderm. The latter is characterised by a sequence of events whereby epithelial ectodermic cells acquire a migratory phenotype, which directly parallels the formation of invasive behaviour in carcinomas. MATERIAL AND METHODS: The present review is based on articles published in well-recognized journals of high international ranking. Some of the considerations also draw on the authors' personal experience in clinical work and basic research. RESULTS: Gastrulation is the process in which the three types of tissue stem cells move to different areas in the embryo (morphogenetic movement) and form the basis for various tissues and organs. This overview calls attention to the fact that events during carcinoma development, are strikingly similar to cellular changes during gastrulation. INTERPRETATION: A thorough understanding of gastrulation may provide a fruitful framework for new insight into cancer cell invasion and metastasis. This knowledge may in turn be exploited to develop drugs with anti invasive properties which could revolutionise the treatment of carcinoma of the prostate and other sites. PMID- 18511971 TI - [Infertility treatment in men with Klinefelter syndrome]. AB - BACKGROUND: Klinefelter syndrome is the most common chromosomal aberration in men with azoospermia. The revision of the Norwegian biotechnology law in 2004 allows infertile couples with this diagnosis to become genetic parents. Surgical sperm extraction has to be performed to enable identification of possible sperm production. The preliminary approval to use this procedure (stated in the biotechnology law) was extended in 2007. This paper presents a short overview of the infertility investigation and treatment in men with Klinefelter syndrome. MATERIAL AND METHODS: The paper is based on more than 10 years of clinical experience in workup and treatment of infertile couples with the diagnosis Klinefelter syndrome. In addition, a non-systematic search in Pubmed and Medline has been carried out. RESULTS: Use of a microscopy during testicular sperm extraction increases the possibility of finding sperm in men with Klinefelter syndrome. The microsurgical procedure also reduces the risk of complications. If the diagnostic recovery has identified testicular sperm, there is a high predictive value of finding sperm in a subsequent surgical procedure. INTERPRETATION: The optimal method for successful sperm recovery in men with Klinefelter syndrome implies the use of microsurgery. This surgical method should therefore be considered established in Norway. PMID- 18511972 TI - [Normal and deviating puberty in boys]. AB - BACKGROUND: Onset of puberty in boys is more complex than in girls, and delayed onset is the most common puberty complication in boys. This article presents the physiology of normal development of male puberty and the background for commonly associated disturbances. MATERIAL AND METHOD: The article builds on clinical experience and relevant publications within pediatric endocrinology. RESULTS AND INTERPRETATION: Mechanisms involved in pubertal development of gonads remain unclear despite intensive research. Height growth as well as the age for onset of puberty are influenced by environmental factors. Genetic factors are however more important determinants within a defined population and one usually inherits the probability for both early and delayed puberty. Gonadotropin releasing hormone (GnRH) neurons in the hypothalamus secrete GnRH in intermittent pulses to the pituitary glands that respond with pulsatile LH and FSH production. These neurons are thus decisive for testicle activity and therefore puberty development. GnRH neurons are inactive during childhood because many types of hypothalamic neurons suppress them. Puberty starts when this suppression is reduced and kisspeptin producing neurons stimulate GnRH neuron activity. At a testicle volume of 4 mL the Leydig cells' testosterone production has reached such a level that pubertal changes become apparent. Delayed or incomplete puberty sometimes occurs in certain syndromes, and complete lack of puberty can also be syndrome-related. Klinefelter's syndrome is associated with gonad dysgenesis expressed as gradual reduction of gonadal function starting after puberty. Cancer treatment during childhood; especially radiation therapy of the gonads, may cause hypogonadism and infertility. It is therefore essential to follow gonad function closely in these patients. In conclusion, each doctor treating children should be able to evaluate the degree of puberty development and when needed request adequate laboratory tests. PMID- 18511973 TI - [Pregnancy in diabetes among Asian, African and Norwegian women]. AB - BACKGROUND: We wanted to describe diabetes (prevalence and types) and pregnancy outcomes among women with diabetes in a multiethnic population. MATERIAL AND METHODS: 509 pregnancies (231 from Asia/Africa and 278 from Norway) were assessed in a population-based, descriptive study of pregnancy with diabetes in Oslo 1993 1998. The outpatient clinic at Aker University Hospital provided information about pregnancies with diabetes and Statistics Norway provided population data. Data were analysed with frequency-analyses and cross-tables. RESULTS: The proportion of pregnancies with diabetes among women 15-49 years was seven times higher in the Asia/Africa group (1.8%, 95% CI = 1.6-2.1) than in the group of ethnic Norwegians (0.3%, 95% CI = 0.2-0.3). Gestational diabetes made up 80% of the diabetes cases among women born in Asia/Africa (13% of type 2 and 5% of type 1) and 48% of the cases among women born in Norway (5% were type 2 and 45% were type 1). In the group with manifest diabetes we found a body mass index > 30 kg/m2 in 42% for women from Asia/Africa and 12% for those from Norway. The percentage of Caesarean section was 37% in the Asia/Africa-group and 42% in the Norway-group. Almost 10% of the newborns in both groups had a birth weight > 4500 g. A postpartum glucose intolerance test for women with gestational diabetes showed that 26% of the women from Asia/Africa had type 2 diabetes and 12% of those from Norway. Information was missing for 43% of those from Asia/Africa and 35% of those from Norway. INTERPRETATION: Diabetes in pregnancy is much more prevalent in women from Asia/Africa than in women born in Norway. Women from Asia/Africa and Norway had different diabetes profiles. High birth weight and interventions in labour were frequent in both groups. Many with gestational diabetes had diabetes type 2. PMID- 18511974 TI - [Serenoa repens in benign prostatic hyperplasia]. AB - Serenoa repens is one of many herbal products used to treat benign prostatic hyperplasia. The treatment has been studied extensively, but the methodological quality has often been poor. Metaanalysis of early studies indicate that the treatment may have favourable effects on patients with benign prostatic hyperplasia, but more recent investigations of better methodological quality have questioned these results. The available documentation does not support use of products containing serenoa repens for these patients. Serenoa repens is associated with mild adverse effects comparable to that of placebo. PMID- 18511975 TI - [Why do we have large intestine?]. PMID- 18511977 TI - [Speculative survey about emergency telephone number 113]. PMID- 18511978 TI - [Bilingualism good]. PMID- 18511980 TI - [No effect of organized cervix cancer screening]. PMID- 18511981 TI - [Stroke experiences--eight years too early]. PMID- 18511984 TI - [Turkish physicians and human rights]. PMID- 18511986 TI - In vitro alpha-glucosidase and alpha-amylase enzyme inhibitory effects of Andrographis paniculata extract and andrographolide. AB - There has been an enormous interest in the development of alternative medicines for type 2 diabetes, specifically screening for phytochemicals with the ability to delay or prevent glucose absorption. The goal of the present study was to provide in vitro evidence for potential inhibition of alpha-glucosidase and alpha amylase enzymes, followed by a confirmatory in vivo study on rats to generate a stronger biochemical rationale for further studies on the ethanolic extract of Andrographis paniculata and andrographolide. The extract showed appreciable alpha glucosidase inhibitory effect in a concentration-dependent manner (IC(50)=17.2+/ 0.15 mg/ml) and a weak alpha-amylase inhibitory activity (IC(50)=50.9+/-0.17 mg/ml). Andrographolide demonstrated a similar (IC(50)=11.0+/-0.28 mg/ml) alpha glucosidase and alpha-amylase inhibitory activity (IC(50)=11.3+/-0.29 mg/ml). The positive in vitro enzyme inhibition tests paved way for confirmatory in vivo studies. The in vivo studies demonstrated that A. paniculata extract significantly (P<0.05) reduced peak blood glucose and area under curve in diabetic rats when challenged with oral administration of starch and sucrose. Further, andrographolide also caused a significant (P<0.05) reduction in peak blood glucose and area under the curve in diabetic rats. Hence alpha-glucosidase inhibition may possibly be one of the mechanisms for the A. paniculata extract to exert antidiabetic activity and indicates that AP extract can be considered as a potential candidate for the management of type 2 diabetes mellitus. PMID- 18511987 TI - Antigen levels of urokinase-type plasminogen activator receptor and its gene polymorphism related to microvessel density in colorectal cancer. AB - We determined the distribution of genotypes and frequencies of alleles of the (CA)(n) repeat polymorphism in intron 3 of the urokinase plasminogen activator receptor (uPAR) gene, uPAR antigen levels and microvessel density (MVD) in tumour and distant mucosa samples from 52 patients with colorectal cancer. The uPAR level was higher for patients with high MVD comparing to patients with lower MVD which may suggest that uPAR can be correlated with progression of colorectal cancer. The significant relationship between the high MVD and uPAR antigen level appeared to be independent of the (CA)(n) repeat polymorphism because no differences in the level of uPAR antigen between carriers of alleles were found. The received results, indicate that uPAR might be considered as a target in colorectal cancer patients' therapy. PMID- 18511988 TI - [The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement: guidelines for reporting of observational studies]. AB - Much of biomedical research is observational. The reporting of such research is often inadequate, which hampers the assessment of its strengths and weaknesses and of a study's generalizability. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Initiative developed recommendations on what should be included in an accurate and complete report of an observational study. We defined the scope of the recommendations to cover three main study designs: cohort, case-control, and cross-sectional studies. We convened a 2-day workshop in September 2004, with methodologists, researchers, and journal editors to draft a checklist of items. This list was subsequently revised during several meetings of the coordinating group and in e-mail discussions with the larger group of STROBE contributors, taking into account empirical evidence and methodological considerations. The workshop and the subsequent iterative process of consultation and revision resulted in a checklist of 22 items (the STROBE Statement) that relate to the title, abstract, introduction, methods, results, and discussion sections of articles. Eighteen items are common to all three study designs and four are specific for cohort, case-control, or cross-sectional studies. A detailed Explanation and Elaboration document is published separately and is freely available on the web sites of PLoS Medicine, Annals of Internal Medicine, and Epidemiology. We hope that the STROBE Statement will contribute to improving the quality of reporting of observational studies. PMID- 18511992 TI - The use of mental health services among young adults with emotional and behavioural problems: equal use for equal needs? AB - OBJECTIVE: Mental health problems are highly prevalent in young adults. Despite possibilities for effective treatment, only about one-third of young adults with mental health problems seek professional help. Little knowledge exists of which groups of young adults are underusing mental health services and for what reasons. The present study examined socio-demographic inequalities in the use of mental health services by young adults, and examined whether such inequalities were attributable to differences in objective need, subjective need, predisposing or enabling factors. DESIGN: Cross-sectional study among the general population aged 19-32 years (2,258 respondents). A postal survey was administered including questions on socio-demographic factors and mental health service use. Data were analyzed with logistic regression analysis. SETTING: South-West Netherlands. PARTICIPANTS: All respondents with serious internalizing and externalizing problems (n = 367). MAIN OUTCOME MEASURE: Twelve-month primary and specialty mental health services use. RESULTS: Only 34.6% of young adults with psychopathology had used any mental health services: 16.2% had used only primary mental health services and 18.4% had used specialty mental health services. No socio-demographic differences were found in the use of only primary mental health services. However, recipients of specialty mental health services were more often female (OR = 2.12, 95% CI = 1.14-3.96), economically inactive (OR = 3.12, 95% CI = 1.59-6.09) or students (OR = 2.38, 95% CI = 1.05-5.42) and they were less often higher educated (OR = 0.49, 95% CI = 0.25-0.97). The higher odds ratio for specialty service use among young adults who were female or economically inactive attenuated when adjusting for need for care. The other socio-demographic disparities in specialty service use did not attenuate when adjusting for need, enabling or predisposing factors. CONCLUSION: Among young adults, equal use of specialty mental health services for equal needs has not been achieved. The underserved groups of young adults oppose the traditionally underserved groups in the general population, and may inform interventions aimed at improving young people's help-seeking behaviours. PMID- 18511993 TI - Age-sex differences in medicinal self-poisonings: a population-based study of deliberate intent and medical severity. AB - BACKGROUND: Deliberate self-harm (DSH) is related to suicide and DSH repetition is common. DSH hospital presentations are often self-poisonings with medicinal agents. While older age and male sex are known risk factors for suicide, it is unclear how these factors are related to the nature and severity of medicinal self-poisoning (SP). Such knowledge can guide prevention strategies emphasizing detecting and treating mental illness and controlling access to means. METHODS: Medicinal SP presentations by 18,383 residents of Ontario, Canada, aged 12 years and older, who presented to a hospital emergency department in that province between April 1, 2001-March 31, 2002 were characterized by the agents taken, identification of deliberate intent and medical severity. RESULTS: We found distinct age-sex differences in the nature and severity of medicinal SP. In youths, aged 12-17, about 40% of presentations involved analgesics, typically not prescribed and most often the acetaminophen agent-group. Females aged 12-64 were identified as deliberate more often than their male counterparts and this pattern occurred in most agent-groups, even among those who took antidepressants. The acetaminophen agent-group was most consistently associated with medical severity and this effect was strongest among female youths. Although medicinal SP was less frequent in the elderly, these presentations tended to be more medically serious and less often identified deliberate. CONCLUSIONS: The high proportion of medicinal SP in youths involving agents typically not prescribed and the medical severity of the acetaminophen agent-group underscore how prevention strategies must extend beyond controlling access to antidepressants. Despite a higher risk for suicide, males and the elderly may not have their deliberate intent detected and therefore, may not receive appropriate treatment. The emergency department can serve as important link to mental health care and usage patterns can provide feedback about the need for system-level enhancements and DSH surveillance. PMID- 18511994 TI - Monitoring the organotin contamination in the Taihu Lake of China by bivalve mussel Anodonta woodiana. AB - As a part of the pilot study on "Freshwater Mussel Watch", butyltin (monobutyltin, MBT; dibutyltin, DB; tributyltin, TBT), and phenyltins (monophenyltin, MPT; diphenyltin DPT; triphenyltin TPT) were analyzed in soft tissues of 15 bivalve mussels Anodonta woodiana sampled from five separate sites (Huzhou, Xueyan, Dapu, Sansandao, and Wulihu) around the Taihu Lake of China in 2004. The residue of total butyltins and total phenyltins in the all mussels ranged 142-1693 and 3.0-90 ng Sn g(-1) dry weight, respectively. Except for the mussels from Xueyan, DBT and MBT accounted more than 60% of total butyltins in those from other four sites. In contrast, TPT were usually almost 100% of the mussels studied. The present study provides most recent information about the organotin contamination in the Taihu Lake, and suggests Anodonta woodiana can be used as a suitable bioindicator. PMID- 18511995 TI - Flat neoplasia of the colon: CT colonography with CAD. AB - Attention is focusing on testing and developing computer aided detection (CAD) systems to reliably highlight flat polyps and cancers to the reporting radiologist during CT colonography. This review will discuss the clinical relevance of flat colonic neoplasia, describe some of the challenges facing CAD detection algorithms, and review the current CAD literature on this topic. PMID- 18511996 TI - The contractile properties of vaginal myofibroblasts: is the myofibroblasts contraction force test a valuable indication of future prolapse development? AB - Using a specific myofibroblast contraction test, we try to predict future utero vaginal prolapse development in young primiparae women. We compare myofibroblast cultures of the vaginal wall in primiparae women (group 1), young multiparae women (group 2) and older multiparae women (group 3) who were operated on for severe utero-vaginal prolapse. A myofibroblast-mediated collagen gel contraction assay determined a contraction factor that was compared in the three groups of women. The myofibroblasts contraction factor after 24 and 48 hours was significantly higher in group 1 women (2.4 +/- 0.6/4.4 +/- 1.9) compared to group 2 (1.6 +/- 0.3/ 1.8 +/- 0.1) andgroup 3 (1.6 +/- 0.3/1.8 +/- 0.3), but showed no differences in group 1 women without (2.1 +/- 0.5/3.5 +/- 1.9) and with (2.7 +/- 0.6/5.1 +/- 1.7) cystocoele. Vaginal myofibroblasts of young women show better contraction forces than young women with severe utero-vaginal prolapse. The latter have a myofibroblast contraction factor similar to those of older post menopausal women operated for the same condition. PMID- 18511997 TI - Choreathetoid movement as an unusual presentation of subdural haematoma. AB - Subdural haematomas can present with a wide variety of symptoms. An atypical presentation can be movement disorders. The key feature is that the history of onset is more rapid than with neurological conditions such as Parkinson's disease. The symptoms can also be an acute worsening of pre-existing disorder. The case discussed involved an 80 year old woman with bilateral choreathetoid movements attributed to a unilateral chronic subdural haematoma. The haematoma was surgically drained and the symptoms resolved. Sporadic reports of similar cases of movement disorders associated with subdural haematomas exist in the literature, but rarely do unilateral haematomas present with bilateral symptoms. Pressure effects, neurotransmitter abnormality and ischaemia have been postulated as reasons for this type of presentation. Unexplained and sudden movement disorders might warrant imaging to rule out a subdural haematoma as the cause. PMID- 18511998 TI - Cure following gene therapy for recurrent glioblastoma multiforme? AB - We report a patient with glioblastoma multiforme who is alive and disease-free 13 years following aggressive treatment with multiple surgeries, radiotherapy, chemotherapy, and gene therapy. PMID- 18511999 TI - Comparison of CT characteristics of extravertebral cement leakages after vertebroplasty performed by different navigation and injection techniques. AB - OBJECTIVE: This study was intended to assess the results of post-operative CT scans in three groups of patients following percutaneous vertebroplasty (VP) using different navigation and injection methods, in an attempt to explain the radiological characteristics of extravertebral cement leakage with relation to needle placement and focused on the ventral epidural accumulation of bone cement. Furthermore, we have suggested a morphological (and functional) classification of the types of cement leakage. METHODS: Between July 2001 and February 2005, 123 percutaneous VP procedures were performed during 75 sessions in 65 patients for treatment of painful osteoporotic vertebral body compression fractures. These included:- Group I: 28 patients, 33 sessions; 50 right sided unilateral VP under fluoroscopic control with central position of the tip of the needle within the bone marrow. Group II: 27 patients, 28 sessions; 50 bilateral VP under fluoroscopic control with separate cement injections into both "hemivertebrae". Group III: 14 patients, 14 sessions; 23 bilateral VP navigated by frameless stereotaxy (neuronavigation). Needles were positioned strictly into the lateral thirds of the vertebral bodies. Leakages were classified as epidural, foraminal, intradiscal, venous paravertebral, compact extravertebral on the post-operative CT scans, and their frequency was compared in relation to the navigation method and the position of the tip of the needle. RESULTS: Group I: extravertebral cement was detected in 23 patients (82%), and in 35 (70%) of the 50 vertebrae treated (ventral epidural: 23 vertebrae = 46%; intradiscal: 12 vertebrae = 24%; venous paravertebral: 8 vertebrae = 16%; intraforaminal: 7 vertebrae = 14%; and compact extravertebral: 3 vertebrae = 6%). Group II: extravertebral cement was detected in 20 patients (74%), and in 38 (76%) of the 50 vertebrae treated (ventral epidural: 12 vertebrae = 24%; intradiscal: 12 vertebrae = 24%; venous paravertebral: 9 vertebrae = 18%; and foraminal: 1 vertebra = 2%). Group III: extravertebral cement could be detected in 10 patients (71%), and in 10 (43%) of the 23 vertebrae treated (ventral epidural: 3 vertebrae = 13%; intradiscal: 8 vertebrae = 34%; venous paravertebral: 4 vertebrae = 17%). CONCLUSION: The incidence of epidural accumulation of bone cement may be concluded to be closely correlated with the position of the tip of the needle. Centrally injected bone cement may easily invade into the basivertebral system, and the material can then be transferred via these veins toward the ventral epidural space, and result in canal compromise and/or compression of the neural elements. The results of statistical analysis (Chi-square test) revealed that injection of bone cement into the lateral third of the vertebral body significantly decreases the extent of ventral epidural leakage. Therefore, a strictly lateral injection is advised, when the tip of the needle is placed into the lateral third of the vertebral body. Frameless stereotaxy navigation improves achievement of accurate needle placement and decreases the frequency of ventral epidural leakage. It is a safe and very accurate method for positioning of the injecting needles. PMID- 18512000 TI - Cerebellar lesions: is there a lateralisation effect on memory deficits? AB - BACKGROUND: Until recently, neurosurgeons eagerly removed cerebellar lesions without consideration of future cognitive impairment that might be caused by the resection. In children, transient cerebellar mutism after resection has lead to a diminished use of midline approaches and vermis transection, as well as reduced retraction of the cerebellar hemispheres. The role of the cerebellum in higher cognitive functions beyond coordination and motor control has recently attracted significant interest in the scientific community, and might change the neurosurgical approach to these lesions. The aim of this study was to investigate the specific effects of cerebellar lesions on memory, and to assess a possible lateralisation effect. METHODS: We studied 16 patients diagnosed with a cerebellar lesion, from January 1997 to April 2005, in the "Centre Hospitalier Universitaire Vaudois (CHUV)", Lausanne, Switzerland. Different neuropsychological tests assessing short term and anterograde memory, verbal and visuo-spatial modalities were performed pre-operatively. RESULTS: Severe memory deficits in at least one modality were identified in a majority (81%) of patients with cerebellar lesions. Only 1 patient (6%) had no memory deficit. In our series lateralisation of the lesion did not lead to a significant difference in verbal or visuo-spatial memory deficits. FINDINGS: These findings are consistent with findings in the literature concerning memory deficits in isolated cerebellar lesions. These can be explained by anatomical pathways. However, the cross lateralisation theory cannot be demonstrated in our series. The high percentage of patients with a cerebellar lesion who demonstrate memory deficits should lead us to assess memory in all patients with cerebellar lesions. PMID- 18512001 TI - Characterization of the full-length genomic sequences of vesicular stomatitis Cocal and Alagoas viruses. AB - In Brazil and Argentina, vesicular stomatitis (VS) is caused by distinct viral strains serologically related to the classical vesicular stomatitis virus Indiana (VSIV), namely VS Indiana-2 (VSIV-2) and VS Indiana-3 (VSIV-3). Here we describe the full-length genomic sequences and organization of the prototype strains of VSIV-2 Cocal virus (COCV) and VSIV-3 Alagoas virus (VSAV). These viruses showed similar genomic organizations to VSIV field isolates except that the non structural C'/C proteins, markedly conserved throughout the vesiculoviruses, were absent in VSAV. Phylogenetic analyses consistently grouped COCV, VSAV and VSIV in a monophyletic group distinct from VSNJV, supporting the classification of these viruses within the Indiana serogroup. PMID- 18512002 TI - Culturable and molecular phylogenetic diversity of microorganisms in an open dumped, extremely acidic Pb/Zn mine tailings. AB - A combination of cultivation-based and molecular-based approaches was used to reveal the culturable and molecular diversity of the microbes inhabiting an open dumped Pb/Zn mine tailings that was undergoing intensive acid generation (pH 1.9). Culturable bacteria found in the extremely acidic mine tailings were Acidithiobacillus ferrooxidans, Leptospirillum ferriphilum, Sulfobacillus thermotolerans and Acidiphilium cryptum, where the number of acidophilic heterotrophs was ten times higher than that of the iron- and sulfur-oxidizing bacteria. Cloning and phylogenetic analysis revealed that, in contrast to the adjacent AMD, the mine tailings possessed a low microbial diversity with archaeal sequence types dominating the 16S rRNA gene library. Of the 141 clones examined, 132 were represented by two sequence types phylogenetically affiliated with the iron-oxidizing archaea Ferroplasma acidiphilum and three belonged to two tentative groups within the Thermoplasma lineage so far represented by only a few environmental sequences. Six clones in the library were represented by the only bacterial sequence type and were closely related to the well-described iron oxidizer L. ferriphilum. The significant differences in the prokaryotic community structures of the extremely acidic mine tailings and the AMD associated with it highlights the importance of studying the microbial communities that are more directly involved in the iron and sulfur cycles of mine tailings. PMID- 18512003 TI - Total knee arthroplasty for rheumatoid knee with bilateral, severe flexion contracture: report of three cases. AB - The treatment of patients with severe flexion contracture of the rheumatoid knee, deprived of ambulation for long periods of time, is challenging. Based on three cases, we indicate the potential risks of posterior dislocation of the knee after total knee arthroplasty. In this pathological condition, surgeons must carefully select the type of implant in order to avoid this serious complication. We also emphasize the importance of working on disuse muscle atrophy of trunk (back, abdominal) and lower limbs, both of which play an integral role in ambulation. The personality of each rheumatoid patient should be carefully considered when considering surgical and rehabilitation options. PMID- 18512004 TI - On the study of local-stress rearrangements during quasi-static plastic shear of a model glass: do local-stress components contain enough information? AB - We present a numerical study of the mechanical response of a 2D Lennard-Jones amorphous solid under steady quasi-static and athermal shear. We focus here on the evolution of local stress components. While the local stress is usually taken as an order parameter in the description of the rheological behaviour of complex fluids, and for plasticity in glasses, we show here that the knowledge of local stresses is not sufficient for a complete description of the plastic behaviour of our system. The distribution of local stresses can be approximately described as resulting from the sum of localized quadrupolar events with an exponential distribution of amplitudes. However, we show that the position of the center of the quadrupoles is not related to any special evolution of the local stress, but must be described by another variable. PMID- 18512005 TI - Vogt-Koyanagi-Harada disease: headache as an initial manifestation. PMID- 18512006 TI - The role of traditional Chinese medicine in colorectal cancer treatment. AB - Surgery, chemotherapy and radiotherapy have been the mainstay of colorectal cancer treatment. There is however current intense research on traditional Chinese medicine (TCM) as novel or additional treatment methods for colorectal cancer. This article reviews the current use of TCM in colorectal cancer so as to increase the awareness of colorectal surgeons. The pathogenesis of colorectal cancer according to TCM is discussed. TCM has been used successfully during the perioperative period to relieve intestinal obstruction, reduce postoperative ileus and reduce urinary retention after rectal surgery. Good results have been reported in the treatment of the complications of chemotherapy and radiation enterocolitis. Favourable results have also been shown in the use of TCM either alone or in combination with chemotherapy to treat advanced colorectal cancer. Molecular studies have shown some TCM compounds to reduce tumour cell proliferation and induce apoptosis. Although the reported results of TCM have been exciting thus far, problems of lack of consensus on treatment regimes and questions on the reliability, validity and applicability of published studies prevent its widespread use. There is now an urgent need for colorectal surgeons to work with TCM physicians in the continuing research on this 6,000-year-old art so as to realize its full potential for our patients. PMID- 18512008 TI - Emergency Hartmann's procedure: morbidity, mortality and reversal rates among Asians. AB - BACKGROUND: Hartmann's procedure and its reversal are associated with high cumulative morbidity rates. We assessed the outcomes of emergency Hartmann's procedures and permanent stoma rates among Asians. METHODS: A retrospective review of all emergency Hartmann's procedure performed from 1996 to 2001 was performed. RESULTS: Emergency Hartmann's procedure was performed in 98 patients with a median age of 68 years. These included 58 patients with cancer and 18 with diverticulitis; 77 patients had pre-existing comorbidities. The mortality and morbidity rates were 19% and 65%, respectively. Mortality was impacted by pre existing respiratory or cardiac disease, age and ASA grade. Likewise, morbidity was significantly increased in the presence of comorbidities, including hypertension and cardiac disease, and age. On multivariate analysis, only age (p=0.003, OR=1.171) and respiratory disease (p=0.029, OR=11.05) affected mortality rates whilst hypertension (p=0.011, OR=5.85) and cardiac disease (p=0.044, OR=5.46) affected morbidity rates. Re-anastomosis was performed in 70 patients, and in 28 of these patients (40%) bowel continuity was re-established after a median of 9 months with a 7% morbidity rate. Reversibility was only related to patient age (p=0.011). CONCLUSIONS: Hartmann's procedure is valid among Asians, and its mortality and morbidity rates are related to patients' pre existing health conditions. The predominant cause is colorectal cancer and permanent stoma rates are related to patient age. PMID- 18512007 TI - Postoperative complications after procedure for prolapsed hemorrhoids (PPH) and stapled transanal rectal resection (STARR) procedures. AB - Procedure for prolapsing hemorrhoids (PPH) and stapled transanal rectal resection for obstructed defecation (STARR) carry low postoperative pain, but may be followed by unusual and severe postoperative complications. This review deals with the pathogenesis, prevention and treatment of adverse events that may occasionally be life threatening. PPH and STARR carry the expected morbidity following anorectal surgery, such as bleeding, strictures and fecal incontinence. Complications that are particular to these stapled procedures are rectovaginal fistula, chronic proctalgia, total rectal obliteration, rectal wall hematoma and perforation with pelvic sepsis often requiring a diverting stoma. A higher complication rate and worse results are expected after PPH for fourth-degree piles. Enterocele and anismus are contraindications to PPH and STARR and both operations should be used with caution in patients with weak sphincters. In conclusion, complications after PPH and STARR are not infrequent and may be difficult to manage. However, if performed in selected cases by skilled specialists aware of the risks and associated diseases, some complications may be prevented. PMID- 18512009 TI - Predictive factors for conversion in laparoscopic colorectal surgery. AB - BACKGROUND: Although laparoscopic colon and rectal surgery can be safely performed in the hands of well-trained surgeons, criteria for patient selection should be further developed in order to decrease the conversion rate. The main objective of this study was to identify predictive factors for conversion of laparoscopic colorectal surgery to an open procedure based on statistical analysis. METHODS: A retrospective survey was performed using data collected from 400 patients who underwent laparoscopic colorectal surgery between March 2000 and December 2006. As potential predictive factors for conversion, we considered demographic characteristics, surgery-related variables and disease-related variables. Univariable analysis was performed to identify individual predictive risk factors for conversion. Factors with p values below 0.05 were included in a regression model. RESULTS: Conversion to open surgery was required in 51 patients (12.7%). Age (>65 years) was the only independent predictive demographic factor (OR=2.3; 95% CI, 1.25-4.46). Low anterior resection (OR=3.9; 95% CI, 1.64-9-18) and complicated diverticulitis (OR=3.9; 95% CI, 1.64-9.18) were also predictive factors. The only predictive factor evidenced in the multivariate analysis was complicated diverticulitis (OR=159.99; 95% CI, 41.02-624.02). Indications for conversion were: adhesions in 53% of the patients, technical problems in 18%, bleeding in 1%, and other indications for the remaining 28%. CONCLUSION: Complicated diverticulitis or cancer of the rectum treated by low anterior resection have higher probabilities of conversion. PMID- 18512010 TI - Differential expression of pro-inflammatory cytokines and a pro-apoptotic protein in pelvic ileal pouches for ulcerative colitis and familial adenomatous polyposis. AB - BACKGROUND: Pouchitis after total rectocolectomy is among the most common complications of patients with ulcerative colitis (UC). However, its frequency is quite rare in patients with familial adenomatous polyposis (FAP). We evaluated the inflammatory and pro-apoptotic activity in endoscopically normal mucosa of the ileal pouch in patients with UC and FAP. METHODS: Twenty patients (10 with UC and 10 with FAP) with "J" pouch after total proctocolectomy were studied as were 10 normal controls. Biopsies were obtained from the mucosa of the pouch of UC and FAP patients and from the normal ileum of controls. The expression levels of TNF alpha, IL-1beta, IL-6, IL-8 and phospho-BAD were determined by immunoblotting. Activated NFkappaB was evaluated by immuno-precipitation and immunoblotting for IkappaB kinase beta. RESULTS: Patients with UC had higher levels of IL-1beta, IL 6, IL-8 and TNF-alpha than patients with FAP. The level of TNF-alpha was higher in patients with UC than in patients with FAP; both patient groups had TNF-alpha levels higher than controls. Activation of NFkappaB was similar in all three groups. The expression of phospho-BAD was significantly lower in patients with FAP than in patients with UC. CONCLUSIONS: As compared with patients with FAP, patients with UC presented increased levels of some pro-inflammatory cytokines, even in the absence of clinical or endoscopic signs of pouchitis. Patients with FAP presented lower levels of pro-inflammatory proteins and of phospho-BAD. These findings may explain the higher rates of progression to pouchitis in UC patients, which could correlate with mucosal atrophy that occurs in inflamed tissue. PMID- 18512011 TI - Distribution of lymph nodes in the mesorectum: how deep is TME necessary? AB - BACKGROUND: Standardization of total mesorectal excision (TME) had a great impact on decreasing local recurrence rates for the treatment of rectal cancer. However, exact numbers and distribution of lymph nodes (LN) along the mesorectum remains controversial with some studies suggesting that few LNs are present in the distal third of the mesorectum. METHODS: Eighteen fresh cadavers without a history of rectal cancer were studied. The rectum was removed by TME and then was divided into right lateral, posterior and left lateral sides, which were further subdivided into 3 levels (upper, middle and lower). A pathologist determined the number and sizes of the LNs in each of the nine areas, b linded to their anatomical origin. RESULTS: Overall, the mesorectum had a mean of 5.7 LNs (SD=3.7) and on average each LN had a maximum diameter of 3.0 mm (SD=2.7). There was no association between the mean number or size of LNs with gender, BMI, or age. There was a significantly higher prevalence of LNs in the posterior location (2.8 per mesorectum) than in the two lateral locations (0.8 and 1.2 per mesorectum; p=0.02). The distribution of LNs in the three levels of the rectum was not significant. CONCLUSIONS: The distribution of LNs reinforces the fact that TME should always include the distal third of the mesorectum. Care must be taken to not violate the posterior aspect of the mesorectum. PMID- 18512012 TI - Prophylactic ureteric catheters in laparoscopic colorectal surgery. AB - BACKGROUND: The purpose of this study was to evaluate the use of ureteric catheter placement in laparoscopic colorectal surgery and to assess the morbidity related to this procedure. METHODS: Between 1994 and 2001, 313 elective laparoscopic colorectal surgeries were performed. Patients with and without ureteric catheters were retrospectively analyzed. RESULTS: Catheter placement was attempted in 149 patients (catheter group) and was not attempted in 164 (controls). There were no significant differences between groups in the number of patients with prior colorectal resection (p=0.286) or other abdominal surgery (p=0.074). Crohn's disease and diverticulitis were more common in the catheter group than among controls (p<0.001). Concomitant intra-abdominal fistula or abscess was present in 29 patients (19.5%) in the catheter group vs. 14 (8.5%) in the control group (p=0.005). The duration of surgery was longer in the catheter group (p=0.001). There were no significant differences in conversion, duration of bladder catheter placement, or length of hospital stay. Urinary tract infection occurred in 3 patients (2.0%) in the catheter group and 7 (4.3%) in the control group (p=0.257) and urinary retention occurred in 3 patients (2.0%) and 11 patients (6.7%), respectively (p=0.045). No intraoperative ureteric injuries occurred in either group. CONCLUSION: Ureteric catheter placement was successful in most cases and was not associated with intraoperative injuries. The increased length of surgery in patients with ureteric catheter placement may attest to the increased severity of pathology in these patients. PMID- 18512013 TI - Comparison of short-and long-term outcomes following either insertion of self expanding metallic stents or emergency surgery in malignant large bowel obstruction. AB - BACKGROUND: Self-expanding metallic stents (SEMS) are now regarded as a safe and effective treatment for an acute obstructing colorectal cancer. SEMS insertion is an invasive procedure that could potentially worsen prognosis. This study assessed the short-and long-term outcomes in patients stented for acute large bowel obstruction and in patients who underwent primary emergency surgery. METHODS: We retrospectively identified 19 patients who underwent SEMS insertion and 23 patients who had primary emergency surgery for left-sided large bowel obstruction as the first presentation of colorectal cancer. RESULTS: There were no significant differences between the 19 patients in the SEMS group and the 23 patients in the primary emergency surgery group in terms of demographics and tumour location and stage. Stent insertion was successful in 16 patients (84%). One patient died from a stent-related perforation and another had a stoma fashioned for stent migration. Stents were a definitive procedure in 2 patients with advanced disease and acted as a "bridge to surgery" in the remaining 12 patients. Compared to the primary surgery group, there was a trend towards a higher primary anastomosis rate in the SEMS group (p=0.08); there were no significant differences in length of hospital stay, 30-day mortality or complication rates between the groups. Long-term prognosis (estimated 3-year survival) did not differ significantly between the groups (p=0.54); this persisted when only curative resections were considered (p=0.80). CONCLUSIONS: Preoperative stent insertion is a safe and effective treatment for large bowel obstruction, and may result in a higher primary anastomosis rate. Stent insertion does not seem to have a deleterious effect on prognosis. PMID- 18512014 TI - Rectal obstruction due to endometriosis. AB - Although endometriosis is a disorder commonly found in reproductive-age women, it does not involve the bowel very often. The circumferential involvement of the rectum is rare and the obstructive symptoms can be difficult to differentiate from those of inflammatory or malignant diseases. We report two patients with rectal endometriosis whose first prominent symptoms were those of intestinal obstruction. A 26-year-old woman was admitted with obstructive symptoms. In order to alleviate the obstruction and extend the preoperative evaluation, a decompressive colostomy was done. The diagnosis of endometriosis was made by laparoscopy and biopsies of the thickened cul-de-sac peritoneum. Another woman, 40 years of age, was referred to us with a colostomy. She had undergone a laparotomy due to an obstructive acute abdomen a year before, and a frozen pelvis was found. Biopsy specimens had been collected and the pathological report revealed endometrioma. A rectosigmoidectomy, encompassing the stenotic rectal segment, was done along with primary anastomosis. The pathological examination confirmed rectal endometriosis. The conclusion is that, although rare, rectal endometriosis can cause significant stenosis of the organ, leading to obstructive symptoms. Despite its low frequency, it should always be considered in the differential diagnosis of rectal stenosis involving women of childbearing age. PMID- 18512015 TI - Rectal diverticulum: a new complication of STARR procedure for obstructed defecation. AB - We report a case of rectal diverticulum developed after stapled transanal rectal resection (STARR) procedure for obstructed defecation. A 21-year-old woman with chronic constipation was diagnosed with a rectocele at defecography. The patient underwent STARR procedure. Six months later, she presented with severe constipation requiring enemas and a worse condition than that preoperatively. Defecography and rectoscopy revealed a rectal wall diverticulum cavity with incomplete elimination of barium enema. The patient underwent transanal diverticulectomy and direct rectal wall repair. STARR procedure can produce new and difficult-to-treat complications and should be reserved for expert colorectal surgeons with proved familiarity in transanal surgery. PMID- 18512016 TI - Laparoscopic caecodivision ACE (antegrade continence enema) procedure. AB - Since the first description of the antegrade colonic enema (ACE) procedure by Malone in 1990, multiple modifications have been developed to overcome some of the initial problems. We describe what we believe to be the first laparoscopic caecodivision method of the ACE procedure. This combines the advantages of laparoscopic technique with those of not using the appendix. PMID- 18512017 TI - Prospective trial evaluating new circular and linear stapler devices for gastrointestinal anastomosis: preliminary data. AB - Several commercial models of stapler devices are available. This study evaluated the ease of use, effectiveness and safety of new commercial stapling devices for gastrointestinal anastomosis. A total of 11 patients (5 men) requiring surgical therapy for benign or malignant disease of the digestive tract were recruited between July and October 2006. Eleven patients were treated with KYGW circular stapler or KYFB linear stapler (Changzhou Kangdi Medical Stapler). In these patients, 14 staplers were used and 21 stapled sutures (16 linear, 5 circular) were performed. Number of anastomoses successfully completed, postoperative anastomotic fistula or dehiscence, days to take fluid and normal diet, length of hospital stay and anastomotic stenosis were recorded. A 10-point questionnaire enquiring about the instrument and anastomotic features was administered to surgeons immediately after the operation in the study group and in 10 control patients treated with standard CDH circular and SDH linear staples (Ethicon Endo Surgery). Mean scores on the questionnaire for the experimental and control groups were good (>7.5) and did not significantly differ for handling, closing ease, bleeding, and overall satisfaction. No case of intra-abdominal sepsis, leakage or intestinal obstruction was recorded in the study group. In the 5 patients with colorectal anastomosis, the anastomotic lumen at 15 days was wide open and at 3 months there were no strictures. These new instruments are valuable for performing gastrointestinal anastomosis and are in conformity with clinical requirements; their use is simple and seems to be safe. PMID- 18512018 TI - Re: Salvaging a linear staple line defect in ultra-low anterior resection. PMID- 18512019 TI - Transvaginal low anterior resection for rectal cancer. AB - Adenocarcinoma of the lower rectum can be resected with a sphincter-sparing procedure but exposure of the lower pelvis may be difficult and sphincter function may be compromised. We have performed a low anterior resection for rectal cancer in a 69-year-old woman with mobilisation of the tumour and anastomosis performed transvaginally without a covering stoma. This way we could get good exposure of the lower rectum and anal sphincters. The patient made an uneventful recovery and was fully continent after surgery. Transvaginal low anterior resection is an alternative route which may be useful in cases of difficult exposure of low rectal cancer. PMID- 18512020 TI - Resolution of secondary alcohols via Carica papaya lipase-catalyzed enantioselective acylation. AB - The Carica papaya lipase-catalyzed acylation of benzylcarbinols with vinyl hexanoate proceeded smoothly and enantiospecifically (E > 200), affording the R esters and leaving the S-alcohols intact. Thus, this plant lipase proved to be a promising biocatalyst for the resolution of alcohols as well as for that of carboxylic acids reported earlier. PMID- 18512021 TI - L-Ribulose production from L-arabinose by an L-arabinose isomerase mutant from Geobacillus thermodenitrificans. AB - Geobacillus thermodenitrificans, with a double-site mutation in L-arabinose isomerase, produced 95 g L-ribulose l(-1 ) from 500 g L-arabinose l(-1) under optimum conditions of pH 8, 70 degrees C, and 10 units enzyme ml(-1) with a conversion yield of 19% over 2 h. The half-lives of the mutated enzyme at 70 and 75 degrees C were 35 and 4.5 h, respectively. PMID- 18512022 TI - Heterologous production of secondary metabolites as pharmaceuticals in Saccharomyces cerevisiae. AB - Heterologous expression of genes involved in the biosynthesis of various products is of increasing interest in biotechnology and in drug research and development. Microbial cells are most appropriate for this purpose. Availability of more microbial genomic sequences in recent years has greatly facilitated the elucidation of metabolic and regulatory networks and helped gain overproduction of desired metabolites or create novel production of commercially important compounds. Saccharomyces cerevisiae, as one of the most intensely studied eukaryotic model organisms with a rich density of knowledge detailing its genetics, biochemistry, physiology, and large-scale fermentation performance, can be capitalized upon to enable a substantial increase in the industrial application of this yeast. In this review, we describe recent efforts made to produce commercial secondary metabolites in Saccharomyces cerevisiae as pharmaceuticals. As natural products are increasingly becoming the center of attention of the pharmaceutical and nutraceutical industries, such as naringenin, coumarate, artemisinin, taxol, amorphadiene and vitamin C, the use of S. cerevisiae for their production is only expected to expand in the future, further allowing the biosynthesis of novel molecular structures with unique properties. PMID- 18512023 TI - The role of DNA synthesis imaging in cancer in the era of targeted therapeutics. AB - Non-specific targets such as DNA and microtubules have been the mainstay of cancer therapeutics and the most effective clinical agents until a decade ago. Advances in genetics, molecular and cellular biology over the past decade led to the development of a new generation of agents that are far more specific and effective. In contrast to progress seen with therapeutic agents, general monitoring targets such as proliferation imaging are just gaining momentum and targeted imaging is still in its infancy. In these paradoxical times, this review assesses the role of proliferation imaging in monitoring the efficacy of targeted therapeutics. PMID- 18512025 TI - Abnormal cytokine production by bone marrow stromal cells of multiple myeloma patients in response to RPMI8226 myeloma cells. AB - INTRODUCTION: Recent studies indicate that bone marrow stromal cells (BMSCs) derived from patients with multiple myeloma (MM) differ from those of healthy donors in their expression of extracellular matrix compounds and in cytokine production. It is not known whether these abnormalities are primary or are acquired by BMSCs on contact with MM cells. MATERIALS AND METHODS: Interleukin (IL)-6, IL-11, IL-10, and tumor necrosis factor (TNF)-alpha production by CD166+ mesenchymal BMSCs and the CD38+/CD138+ RPMI8226 myeloma cell line cultivated in vitro in monocultures or co-cultivated under cell-to-cell contact or non-contact conditions in the presence of a tissue culture insert were measured. Intracellular cytokines were measured by flow cytometry analysis as the percentage of cytokine-producing cells or by mean fluorescence intensity as the level of cytokine expression in cells. Additionally, ELISA was used to measure IL 6, soluble IL-6 receptor (sIL-6R), IL-11, IL-10, TNF-alpha, B-cell-activating factor of the TNF family (BAFF), hepatocyte growth factor (HGF), and osteopontin (OPN) production in the supernatants of the cultures and co-cultures. RESULTS: A higher ability of the BMSCs of MM patients than in controls was detected to produce IL-6, IL-10, TNF-alpha, OPN, and especially HGF and BAFF in response to the RPMI8226 cells. Moreover, the BMSCs of the MM patients significantly enhanced the production of sIL-6R by the RPMI8226 cells. DISCUSSION: Cytokines over expressed by BMSCs of MM patients can function as growth factors for myeloma cells (IL-6, IL-10, HGF), migration stimulatory factors for tumor plasma cells (TNF-alpha, HGF), adhesion stimulatory factors (HGF, BAFF and OPN), stimulators of osteoclastogenesis (IL-6, TNF-alpha), and angiogenic factors (TNF-alpha). The results of this experiment strongly suggest that the BMSCs from MM patients differed in spontaneous and myeloma cell-induced production of cytokines, especially of HGF and BAFF, and these abnormalities were both primary and acquired by the BMSCs on contact with the MM cells. This in turn suggests the presence of an undefined, autocrine stimulation pathway resulting in a prolonged production of cytokines even in long-term cultures in vitro and in vivo. These abnormalities might provide optimal conditions for the proliferation and differentiation of residual tumor cells or their precursors in the affected bone marrow. PMID- 18512026 TI - Prevalence of human herpesvirus 6 antibodies and DNA in allogeneic stem cell transplant patients: two-year single centre experience. AB - INTRODUCTION: Human herpesvirus 6 (HHV-6) has been recognized as a potentially significant pathogen in hemopoietic stem cell transplant (HSCT) recipients. Different clinical manifestations have been described, including fever, skin rash, bone marrow suppression, and encephalitis. MATERIALS AND METHODS: A retrospective review of a group of 26 adult recipients of allogeneic HSCTs was conducted. Serum samples taken before transplant were examined for the presence of specific anti-HHV-6 IgM and IgG antibodies. After transplantation, quantitative real-time PCR was used to determine viral load in plasma samples from days 0-180 post-transplant. RESULTS: HHV-6 DNA was detected in plasma samples in 8 (30%) of the 26 recipients between days 18 and 40 after transplantation. All of them developed fever of unknown origin and over 50% had graft-versus-host disease features. Three individuals from this group died during detectable HHV-6 viremia. Another two recipients showed a single positive PCR result at a later time. Infection with HHV-6 was thus confirmed in 10 (38.5%) of the 26 graft recipients. CONCLUSIONS: There is a high frequency of detectable HHV 6 viral load in stem cell transplant recipients in Poland. Further investigation to monitor HHV-6 reactivation in graft recipients will be important to improve outcome for these patients. PMID- 18512027 TI - Dual use and the ethical responsibility of scientists. AB - The main normative problem in the context of dual use is to determine the ethical responsibility of scientists especially in the case of unintended, harmful, and criminal dual use of new technological applications of scientific results. This article starts from an analysis of the concepts of responsibility and complicity, examining alternative options regarding the responsibility of scientists. Within the context of the basic conflict between the freedom of science and the duty to avoid causing harm, two positions are discussed: moral skepticism and the ethics of responsibility by Hans Jonas. According to these reflections, four duties are suggested and evaluated: stopping research, systematically carrying out research for dual-use applications, informing public authorities, and not publishing results. In the conclusion it is argued that these duties should be considered as imperfect duties in a Kantian sense and that the individual scientist should be discharged as much as possible from obligations which follow from them by the scientific community and institutions created for this purpose. PMID- 18512024 TI - Cancer stem cells as targets for cancer therapy: selected cancers as examples. AB - It is becoming increasingly evident that cancer constitutes a group of diseases involving altered stem-cell maturation/differentiation and the disturbance of regenerative processes. The observed malignant transformation is merely a symptom of normal differentiation processes gone astray rather than the primary event. This review focuses on the role of cancer stem cells (CSCs) in three common but also relatively under-investigated cancers: head and neck, ovarian, and testicular cancer. For didactic purpose, the physiology of stem cells is first introduced using hematopoietic and mesenchymal stem cells as examples. This is followed by a discussion of the (possible) role of CSCs in head and neck, ovarian, and testicular cancer. Aside from basic information about the pathophysiology of these cancers, current research results focused on the discovery of molecular markers specific to these cancers are also discussed. The last part of the review is largely dedicated to signaling pathways active within various normal and CSC types (e.g. Nanog, Nestin, Notch1, Notch2, Oct3 and 4, Wnt). Different elements of these pathways are also discussed in the context of therapeutic opportunities for the development of targeted therapies aimed at CSCs. Finally, alternative targeted anticancer therapies arising from recently identified molecules with cancer-(semi-)selective capabilities (e.g. apoptin, Brevinin-2R) are considered. PMID- 18512029 TI - The significance of Treg cells in defective tumor immunity. AB - Regulatory T cells (Treg) enriched in FoxP3+, glucocorticoid-induced TNF receptor+, and cytotoxic T-lymphocyte-associated antigen-4+ exert a potential to suppress effector T cells in the periphery. These cells exist in markedly higher proportions within tumor-infiltrating lymphocytes, peripheral blood lymphocytes, and/or regional lymph node lymphocytes of patients with cancer and their frequencies are suggested to be strongly related to tumor progression and inversely correlated with the efficacy of treatment. Tumor-specific Treg cells require ligand-specific activation and cell-to-cell contact to exert their suppressive activity on tumor-specific effector cells (CD8+ cytotoxic T lymphocytes and CD4+ Th cells), which includes decreased cytotoxity, proliferation, and Th1 cytokine secrection. Depletion or blockade of Treg cells can enhance immune protection from tumor-associated antigens that are expressed as self antigens. Recent studies revealed that lymphoma T cells might adopt a Treg profile as well. Studies assessing the influence of chemotherapy on Treg cells have also been included in this review. PMID- 18512028 TI - A direct comparison of rejection by CD8 and CD4 T cells in a transgenic model of allotransplantation. AB - INTRODUCTION: The relative contributions of CD4+ and CD8+ T cells to transplant rejection remain unknown. The authors integrated a previous model of CD4-mediated graft rejection with a complementary model of CD8-mediated rejection to directly compare the function of graft-reactive CD4+ and CD8+ lymphocytes in vivo in a model where rejection requires transgenic T cells. These studies allow direct comparison of CD4 and CD8 T cell responses to the same antigen without the confounding effects of T cell depletion or homeostatic proliferation. MATERIALS AND METHODS: Clone 4 and TS1 mice possess MHC class I- and II-restricted CD8+ and CD4+ T cells, respectively, which express transgenic T cell receptors that recognize the influenza hemagglutinin antigen (HA). We compared the in vivo response of CFSE-labeled, HA-specific transgenic CD8+ and CD4+ T cells after adoptive transfer into syngeneic BALB/c mice grafted with HA-expressing skin. RESULTS: As in the authors' CD4+ model, HA104 skin was consistently rejected by both Clone 4 mice (n=9, MST: 14.2) and by 5 x 10(5) Clone 4 lymphocytes transferred to naive BALB/c hosts that do not otherwise reject HA+ grafts. Rejection correlated with extensive proliferation of either graft-reactive T cell subset in the draining lymph nodes, and antigen-specific CD4+ and CD8+ cells acquired effector function and proliferated with similar kinetics. CONCLUSIONS: These data extend the authors' unique transgenic transplantation model to the investigation of CD8 T cell function. The initial results confirm fundamental functional similarity between the CD4 and CD8 T cell subsets and provide insight into the considerable redundancy underlying T cell mechanisms mediating allograft rejection. PMID- 18512031 TI - Automated sequence-specific protein NMR assignment using the memetic algorithm MATCH. AB - MATCH (Memetic Algorithm and Combinatorial Optimization Heuristics) is a new memetic algorithm for automated sequence-specific polypeptide backbone NMR assignment of proteins. MATCH employs local optimization for tracing partial sequence-specific assignments within a global, population-based search environment, where the simultaneous application of local and global optimization heuristics guarantees high efficiency and robustness. MATCH thus makes combined use of the two predominant concepts in use for automated NMR assignment of proteins. Dynamic transition and inherent mutation are new techniques that enable automatic adaptation to variable quality of the experimental input data. The concept of dynamic transition is incorporated in all major building blocks of the algorithm, where it enables switching between local and global optimization heuristics at any time during the assignment process. Inherent mutation restricts the intrinsically required randomness of the evolutionary algorithm to those regions of the conformation space that are compatible with the experimental input data. Using intact and artificially deteriorated APSY-NMR input data of proteins, MATCH performed sequence-specific resonance assignment with high efficiency and robustness. PMID- 18512030 TI - Manipulating B cell homeostasis: a key component in the advancement of targeted strategies. AB - Understanding the homeostatic mechanisms governing lymphocyte pools achieves critical importance as lymphocyte-targeted therapies expand in use and scope. The primacy of B lymphocyte stimulator (BLyS) family ligands and receptors in governing B lymphocyte homeostasis has become increasingly clear in recent years, affording insight into novel opportunities and potential pitfalls for targeted B cell therapeutics. Interclonal competition for BLyS-BR3 interactions determines the size of naive B cell pools and can regulate the stringency of selection applied as cells complete maturation. Thus one of the predicted consequences of ablative therapies targeting primary pools is relaxed negative selection. This suggests that BLyS levels and B cell reconstitution rates may serve useful prognostic roles and that BLyS itself might be targeted to circumvent relapse. Alternatively, manipulations that allow rare, minimally autoreactive specificities to survive and mature may lead to opportunities in cases where antibody-based vaccine development has heretofore been unsuccessful. BLyS family ligands and receptors also play a role in activated and memory B cell pools, suggesting they might likewise be targeted to promote or delete particular antigen-experienced subpopulations in a similar way. PMID- 18512032 TI - Fruit mineral contents of six wild species of the North Andean Patagonia, Argentina. AB - The fruit mineral contents (K, Ca, Ba, Br, Zn, Co, Cr, Fe, Na, Rb, Cs, and Sr) of four native and two exotic naturalized shrubs growing in different areas of the Andean Patagonian region of Argentina were investigated. Native species Berberis darwinii, Berberis microphylla (Berberidaceae), Aristotelia chilensis (Elaeocarpaceae) and Ribes magellanicum (Saxifragaceae) produce small berries while the fruits of the exotic species Rosa rubiginosa and Rosa canina (Rosaceae) are aggregates of aquenes. They are used to prepare jams, tea, liquors, and ice creams. Native shrub fruits had higher content of Br, Zn, Co, Cr, Fe, Mo, and Na than those of the exotic naturalized species. Rosa species showed the highest contents of Ca and Ba in their fruits (the mean content doubled those of the native plant fruits). The fruit nutrient content found in the studied species was similar or higher than other values reported for fruits of temperate and tropical species in the world. PMID- 18512033 TI - Pulmonary hypertension, heart failure and neutropenia due to diazoxide therapy. AB - Primary persistent hyperinsulinaemic hypoglycaemia is characterised by clinical symptoms that occur when blood glucose levels drop below the normal range. Diazoxide treatment remains the mainstay of medical therapy. Tolerance of diazoxide is usually excellent, but several side effects of this drug have been described. We present a 4-month-old girl who developed pulmonary hypertension, heart failure and neutropenia during diazoxide therapy. Diazoxide toxicity was suspected and the drug was withdrawn on day 13. During the next 3 days, respiratory and haemodynamic status dramatically improved and she was weaned from mechanical ventilation. Control white blood cell count was 8800 cells/mm(3) and a new echocardiography showed modreduction of pulmonary artificial pressure to 20 mmHg and resolution of atrial and ventricular enlargement. Paediatric physicians should be in mind of pulmonary hypertension, heart failure and neutropenia developing during diazoxide therapy. PMID- 18512034 TI - [Nicotine in plastic surgery : a review]. AB - BACKGROUND: The surgical complication rate is significantly increased in active smoking patients. However there are no evidence-based recommendations regarding smoking among patients seeking plastic surgical procedures. METHODS: MEDLINE analysis was performed of all relevant clinical and experimental papers from 1965 to 2008. RESULTS: In face-lift operations smokers present a 13-fold risk of skin necrosis. In mamma reduction procedures the risk among smokers is doubled for number of complications, with T-incision site necrosis (odds ratio 3.1) and infection rate (OR 3.3) significantly elevated among active smokers. Transverse rectus abdominis myocutaneous flaps for breast reconstruction are associated with significantly higher flap necrosis rates for smokers than nonsmokers (19% vs 9%, P=0.005). The smoking history can be indicative, but usually the number of cigarettes is drastically underestimated. Cotinine testing is a method of determining smoking quantitatively up to 4 days before testing. CONCLUSION: Four weeks of abstinence from smoking reduces smoking-associated complications. Despite the published evidence, smoking is no longer relevant in the German 2008 Disease-Related Group for plastic surgical procedures. PMID- 18512035 TI - [27-year old pregnant woman with syncope and dyspnea after aortic alloplastic heart valve replacement 15 years ago]. AB - A 27-year old pregnant woman presented in our emergency department with syncope, dyspnea and complaints about an overall impairment. She had received aortic alloplastic heart valve replacement due to a congenital, valvular stenosis in 1993. We diagnosed a prosthetic heart valve thrombosis associated with cardiac decompensation. Due to tachycardia and critical hypotension we decided to perform fibrinolytic therapy with tenecteplase. After fibrinolysis both prosthetic heart valve leaflets were separating in normal and regular function. The patient was initially anticoagulated with low molecular weight heparin, which was switched during the hospital stay to unfractionated heparin. Later oral anticoagulation with phenprocoumon was initiated. At 36 weeks of gestation an uneventful elective abdominal caesarean section was performed. The mother and the newborn were in healthy condition. Hypercoaguability in pregnancy is a serious problem, especially for patients who already have an existing indication for therapeutic anticoagulation. Fibrinolysis should definitely be considered an option during pregnancy in critical and life-threatening situations. PMID- 18512036 TI - [Infrarenal aortic aneurysm]. AB - An abdominal aortic aneurysm is defined as the increase of infrarenal aortic diameter of 3.0 cm and more. Infrarenal aortic aneurysm is frequent in the elderly and causes 1-3% of all deaths among men aged between 65 and 85 years. These aneurysms are typically asymptomatic until the life threatening event of rupture. Therefore screening of risk populations like elderly persons and persons with cardiovascular risk factors for aortic aneurysm seems to be most important. An aortic aneurysm is usually detected by sonography. An infrarenal aortic aneurysm with a diameter of 5.0-5.5 cm should be treated either with open surgical or endovascular therapy. If surgical or endovascular therapy is indicated, additional computer tomography (CT) or magnet resonance imaging (MRI) are necessary. The mode of treatment mainly depends on patient co-morbidity and on morphology of the aneurysm according to the CT/MRI-findings and should be determined individually. PMID- 18512037 TI - [Isolated rupture of the short head of the biceps muscle after a traffic accident]. AB - Isolated rupture of the short head of the biceps muscle is a rare injury and only very few cases have been published. This case study describes an isolated, complete, traumatic rupture through the short head of the biceps muscle in a 39 year-old healthy male who was involved in a car accident. We report on the diagnostic and therapeutic measures and compare the procedure with data from the literature. PMID- 18512038 TI - [Combined dorsal and palmar plate osteosynthesis for intraarticular distal radius fractures]. AB - BACKGROUND: Complex comminuted intraarticular fractures of the distal radius require anatomic restoration. In rare cases, this aim can only be achieved by a combined dorsal and volar plate fixation despite increasing experience with volar locking compression implants. This retrospective quality control study investigated functional, radiological and subjective outcomes of patients treated with this technique. METHODS: Between March 1999 and January 2003, 30 out of 360 patients who were operated on for an unstable distal radius fracture with complex C2/C3 type distal radius fractures at the Division of Trauma Surgery of the University Hospital Zurich, had been treated with a combined dorso-palmar plate fixation (dorsal two 1/4 tubular plates, volar 3.5 mm T-plate) and were included in this study (9 female, 21 male, mean age 52 years). Of the 30 patients 25 could be evaluated at an average of 29 months after injury. RESULTS: Anatomic reconstruction could not be achieved in all cases, 56% showed mild and 28% explicit signs of arthrosis. Flexion reached 66% and extension 75% of the contralateral wrist, whereas pronation reached 98% and supination 91%, respectively. Grip strength achieved 75% of that of the contralateral side and 10 patients (40%) developed a complex regional pain syndrome (CRPS). Return to work was possible 120 days after the injury. CONCLUSION: With the dorso-palmar plate fixation joint reconstruction in complex intraarticular distal radius fractures can be achieved with a satisfactory subjective, functional, and radiologic result, although additional soft tissue injury was caused by the bilateral approach. The patient has to be informed of the high rate of CRPS and the long period of disability. PMID- 18512039 TI - [Intramedullary nail removal in the upper and lower limbs. Should we recommend this operation?]. AB - BACKGROUND: Absolute indications for removing intramedullary locking nails (ILN) are undisputed, but there are also relative indications when implant removal might be discussed. The aim of our study was to evaluate complications of ILN removal in the upper and lower extremities. METHODS: Four hundred sixty (460) patients who underwent interlocking nail removal were reviewed regarding complications after removal of implants in the humerus, femur, or tibia. RESULTS: The most common complications were delayed wound healing and wound infections. For the humerus, the complication rate of implant removals due to absolute indication was 29%, and the rate for removals due to relative indication was 12%. In the forearm, no complications were seen. Patients who underwent ILN removal in the femur or tibia for absolute indication had a 21% complication rate; the complication rate in patients with relative indication was 10%. CONCLUSION: The complication rate of interlocking nail removal is too high to justify such a procedure without clear indication. PMID- 18512040 TI - [Trauma hospital. Knee arthroplasty as primary therapy for proximal tibial fracture]. AB - Ten patients aged 55-85 years with a tibial head fracture AO B3 or C3 were treated primarily by implantation of an endoprosthesis. There were one unilateral, three superficial, and six revision-type prostheses. Follow-up was 6 months to 3 years; two patients were lost to follow-up. There were no intra- or postoperative complications except one deep infection which could be cured by repeated arthroscopic lavage. At last follow-up all eight patients were completely or almost pain free; the extension deficit was less than 10 degrees , and flexion was 100 degrees or more. Primary endoprosthetic replacement of the knee joint is a valuable procedure for the treatment of complex tibial head fractures in elderly patients. PMID- 18512041 TI - Allelic differentiation of Kunitz trypsin inhibitor in wild soybean (Glycine soja). AB - Soybean Kunitz trypsin inhibitor (SKTI) has several polymorphic types, which are controlled by co-dominant multiple alleles at a single locus. Of these types, Tia and Tib are predominant types, and there are nine differences in amino acids between Tia and Tib. Recently, an intermediate transitional type (Tibi5) between them was detected. However, other transitional types have not been detected despite surveys of many cultivated and wild soybeans. One of the reasons why other transitional variants have not been found is inferred to be due to the difficulty of the detection of SKTI protein variants by polyacrylamide gel electrophoresis (PAGE). To detect novel variants of SKTI, nucleotide sequence analysis in addition to PAGE was carried out. Four new variants were found from many Japanese wild soybeans. Of these variants, three (designated as Tiaa1, Tiaa2, Tiab1) were detected through gene sequence analysis on wild soybeans having the same electrophoretic mobility as Tia, and one (Tig) was detected through PAGE. The Tig variant showed a slightly lower electrophoretic mobility than Tic. The nucleotide sequences of Tig were identical to those of Tib except for one T-->C transitional mutation at position +340. The sequences of Tiaa1 and Tiaa2 genes were identical to those of Tia with the exception of a G-->A mutation at position +376 and a T-->C mutation at +404, respectively. The sequence of Tiab1 differed from Tia by three nucleotides: C-->A at position +331, T-->C at +459 and A-->G at +484. Of the three nucleotide changes, two were common to Tiab1, Tibi5 and Tib, suggesting that Tiab1 is an intermediate transitional type between Tia and Tib. Our results suggest that Tib type has been differentiated through a series of mutations from Tia before the domestication of cultivated soybean. PMID- 18512042 TI - Genotyping with real-time PCR reveals recessive epistasis between independent QTL conferring resistance to common bacterial blight in dry bean. AB - Resistance to common bacterial blight in common bean is a complex trait that is quantitatively inherited. Combining QTL is the current strategy for improving resistance, but interactions among different QTL are unknown. We examined the interaction between two independent QTL present in dry bean breeding line XAN 159. The QTL were studied in a near isogenic population consisting of 120 BC6:F2 plants. Each BC6:F2 plant was evaluated for disease reaction at several time points after pathogen inoculation and the dominant SCAR markers linked with QTL on linkage groups B6 (BC420 approximately QTL) and B8 (SU91 approximately QTL) were interpreted as codominant markers using real time PCR assays. This enabled assignment of BC6:F2 plants to all nine possible genotypes. Reaction to CBB in BC6:F2 plants was characterized by an epistatic interaction between BC420 and SU91 such that: 1) the expression of BC420 was epistatically suppressed by a homozygous recessive su91//su91 genotype; 2) SU91//SU91 and SU91//su91 genotypes conditioned an intermediate disease reaction when homozygous recessive for bc420//bc420; and 3) the highest level of disease resistance was conferred by genotypes with at least a single resistance allele at both QTL (BC420//-; SU91// ). Segregation for resistance among BC6:F3 plants derived from BC6:F2 plants that were heterozygous for both QTL did not deviate significantly from expected ratios of 9 resistant: 3 moderately resistant: 4 susceptible. This is consistent with a recessive epistatic model of inheritance between two loci. These results indicate breeders will realize greatest gains in resistance to CBB by selecting breeding materials that are fixed for both QTL. This is a first report of a qualitative digenic model of inheritance discerning an interaction between two QTL conditioning disease resistance in plants. PMID- 18512043 TI - Fuel metabolism during exercise in euglycaemia and hyperglycaemia in patients with type 1 diabetes mellitus--a prospective single-blinded randomised crossover trial. AB - AIMS/HYPOTHESIS: We assessed systemic and local muscle fuel metabolism during aerobic exercise in patients with type 1 diabetes at euglycaemia and hyperglycaemia with identical insulin levels. METHODS: This was a single-blinded randomised crossover study at a university diabetes unit in Switzerland. We studied seven physically active men with type 1 diabetes (mean +/- SEM age 33.5 +/- 2.4 years, diabetes duration 20.1 +/- 3.6 years, HbA1c 6.7 +/- 0.2% and peak oxygen uptake [VO2peak] 50.3 +/- 4.5 ml min(-1) kg(-1)). Men were studied twice while cycling for 120 min at 55 to 60% of VO2peak, with a blood glucose level randomly set either at 5 or 11 mmol/l and identical insulinaemia. The participants were blinded to the glycaemic level; allocation concealment was by opaque, sealed envelopes. Magnetic resonance spectroscopy was used to quantify intramyocellular glycogen and lipids before and after exercise. Indirect calorimetry and measurement of stable isotopes and counter-regulatory hormones complemented the assessment of local and systemic fuel metabolism. RESULTS: The contribution of lipid oxidation to overall energy metabolism was higher in euglycaemia than in hyperglycaemia (49.4 +/- 4.8 vs 30.6 +/- 4.2%; p < 0.05). Carbohydrate oxidation accounted for 48.2 +/- 4.7 and 66.6 +/- 4.2% of total energy expenditure in euglycaemia and hyperglycaemia, respectively (p < 0.05). The level of intramyocellular glycogen before exercise was higher in hyperglycaemia than in euglycaemia (3.4 +/- 0.3 vs 2.7 +/- 0.2 arbitrary units [AU]; p < 0.05). Absolute glycogen consumption tended to be higher in hyperglycaemia than in euglycaemia (1.3 +/- 0.3 vs 0.9 +/- 0.1 AU). Cortisol and growth hormone increased more strongly in euglycaemia than in hyperglycaemia (levels at the end of exercise 634 +/- 52 vs 501 +/- 32 nmol/l and 15.5 +/- 4.5 vs 7.4 +/- 2.0 ng/ml, respectively; p < 0.05). CONCLUSIONS/INTERPRETATION: Substrate oxidation in type 1 diabetic patients performing aerobic exercise in euglycaemia is similar to that in healthy individuals revealing a shift towards lipid oxidation during exercise. In hyperglycaemia fuel metabolism in these patients is dominated by carbohydrate oxidation. Intramyocellular glycogen was not spared in hyperglycaemia. PMID- 18512044 TI - Neural trigger and cycling off during helmet pressure support ventilation: the epitome of the perfect patient ventilator interaction? PMID- 18512046 TI - ACE insertion/deletion polymorphism in sepsis and acute respiratory distress syndrome. PMID- 18512045 TI - Subject-ventilator synchrony during neural versus pneumatically triggered non invasive helmet ventilation. AB - OBJECTIVE: Patient-ventilator synchrony during non-invasive pressure support ventilation with the helmet device is often compromised when conventional pneumatic triggering and cycling-off were used. A possible solution to this shortcoming is to replace the pneumatic triggering with neural triggering and cycling-off-using the diaphragm electrical activity (EA(di)). This signal is insensitive to leaks and to the compliance of the ventilator circuit. DESIGN: Randomized, single-blinded, experimental study. SETTING: University Hospital. PARTICIPANTS AND SUBJECTS: Seven healthy human volunteers. INTERVENTIONS: Pneumatic triggering and cycling-off were compared to neural triggering and cycling-off during NIV delivered with the helmet. MEASUREMENTS AND RESULTS: Triggering and cycling-off delays, wasted efforts, and breathing comfort were determined during restricted breathing efforts (<20% of voluntary maximum EA(di)) with various combinations of pressure support (PSV) (5, 10, 20 cm H(2)O) and respiratory rates (10, 20, 30 breath/min). During pneumatic triggering and cycling-off, the subject-ventilator synchrony was progressively more impaired with increasing respiratory rate and levels of PSV (p < 0.001). During neural triggering and cycling-off, effect of increasing respiratory rate and levels of PSV on subject-ventilator synchrony was minimal. Breathing comfort was higher during neural triggering than during pneumatic triggering (p < 0.001). CONCLUSIONS: The present study demonstrates in healthy subjects that subject ventilator synchrony, trigger effort, and breathing comfort with a helmet interface are considerably less impaired during increasing levels of PSV and respiratory rates with neural triggering and cycling-off, compared to conventional pneumatic triggering and cycling-off. PMID- 18512047 TI - DETA/NONOate, a nitric oxide donor, produces antidepressant effects by promoting hippocampal neurogenesis. AB - RATIONALE: Increasing evidence suggests that depression may be associated with a lack of hippocampal neurogenesis. Our recent study shows that endogenous nitric oxide (NO) contributes to chronic mild stress (CMS)-induced depression by suppressing hippocampal neurogenesis. OBJECTIVES: The aim of this study was to investigate the effects of exogenous NO in CMS-induced depression in young adult mice. RESULTS: In normal mice, administration of a pure NO donor (Z)-1-[N-(2 aminoethyl)-N-(2-ammonioethyl) aminio] diazen-1-ium-1,2-diolate (DETA/NONOate; 0.4 mg/kg, i.p., for 7 days) produced an antidepressant-like effect and significantly increased hippocampal neurogenesis. The mice exposed to CMS exhibited behavioral changes typical of depression and impaired neurogenesis in the hippocampus. Treatment with DETA/NONOate (0.4 mg/kg, i.p., for 7 days) reversed CMS-induced behavioral despair and hippocampal neurogenesis impairment. We treated mice with a telomerase inhibitor 3'-azido-deoxythymidine (AZT; 100 mg/kg, i.p., for 14 days) to disrupt neurogenesis. From day 4 to day 11 of AZT treatment, mice were injected with DETA/NONOate (0.4 mg/kg, i.p., for 7 days). Disrupting hippocampal neurogenesis blocked the antidepressant effect of DETA/NONOate. CONCLUSIONS: Our findings suggest that exogenous NO benefits chronic stress-induced depression by stimulating hippocampal neurogenesis and may represent a novel approach for the treatment of depressive disorders. PMID- 18512049 TI - Motor cortex excitability changes during imagery of simple reaction time. AB - Imagining motor actions is enough to enhance cortical motor excitability. However, the fact that execution of the motor task has to be inhibited should have a correlate on the extent of cortical excitability enhancement. Therefore, we examined the possible differences between real and motor imagery of simple reaction time tasks (SRT) in the facilitation of the motor-evoked potential (MEP) to transcranial magnetic stimulation (TMS) and in the reduction of short-interval intracortical inhibition (SICI) taking place before the movement onset. Thirteen healthy volunteers were requested to perform visual real or imaginary SRT tasks (rSRT and iSRT) with their dominant hand. For rSRT, subjects had to perform a rapid isometric squeeze as soon as possible after the imperative signal. For iSRT they had to imagine the same movement without actually doing it. Electromyographic (EMG) signals were recorded from the first dorsal interosseus (FDI) muscle. The mean EMG onset was calculated for each subject in rSRT trials. Single-and paired-pulse TMS (at an interstimulus interval of 2.5 ms) were applied at rest and at time intervals of -25, -50, -75, -100 and -125 ms before the expected real (rSRT) mean EMG onset. In rSRT there was a significant increase of MEP to single-pulseTMS at the intervals of -50 and -25 ms, and in iSRT at -50 ms in comparison to the rest condition. % SICI changes were significantly reduced at the intervals of -75, -50 and -25 ms, before EMG the onset in rSRT and at -50 and -25 in iSRT (P < 0.05) in comparison to % SICI changes at rest. The differences between MEPs to spTMS and MEP to ppTMS were significantly larger at rest, -125 and -100 ms intervals in rSRT, and at all intervals in iSRT. Imagining to move causes an increase in corticospinal excitability and a decrease in intracortical inhibition that follow a temporal profile similar to those observed with real movements. However, complete removal of SICI happened only in rSRT at the shortest intervals before the EMG onset. Such action may delineate new tool in motor rehabilitation of patients who have limitation to move. PMID- 18512050 TI - Arm movement metrics influence saccade metrics when looking and pointing towards a memorized target location. AB - Saccades are known to influence subsequent arm movements. There is less information to suggest that the characteristics of saccades depend on the reaching movements they accompany. To explore this issue, we studied the systematic errors of saccades generated by two adult female Rhesus monkeys (Macaca Mulata), which were trained to perform center-out saccades and reaching arm movements to the memorized location of targets. The mean error of saccades executed in isolation differed significantly from that of saccades that were executed towards the same target location and accompanied a reaching movement. This difference was observed in both animals whether they used their right or left arm, whether the size of the movement was equal to 10 or 15 degrees and whether there was no delay or a 3 s delay between the extinction of a visual target and the cue to move. Moreover, the endpoints of saccades and those of the arm-reaching movements in the reaching task were significantly correlated. These data suggest that signals specifying the metrics of limb movements influence those specifying the metrics of preceding saccades at a programming stage. PMID- 18512051 TI - Cortical processing of tactile stimuli applied in quick succession across the fingertips: temporal evolution of dipole sources revealed by magnetoencephalography. AB - We used magnetoencephalography (MEG) in 10 healthy human subjects to study cortical responses to tactile stimuli applied to the fingertips of digits 2-5 of the right hand. Each stimulus lasted 50 ms and was produced by air-driven elastic membranes. Four-hundred stimuli were delivered on each finger in three temporal patterns (conditions). In the "Discrete" condition, stimuli were applied to each finger repetitively with an interstimulus interval (ISI) of 1-2 s. In the "Continuous" condition, stimuli were applied to the fingers sequentially as four stimulus trains with zero ISI and 1-2 s intervening between trains. Finally, in the "Gap" condition, stimuli were applied as in the Continuous condition but with an ISI of 50 ms. A sensation of tactile motion across fingers (digit 2 --> digit 5) was reported by all subjects in the Continuous and Gap conditions. Cortical responses were extracted as single equivalent current dipoles over a period of 1 s following stimulus onset. In all three conditions, initial responses in left primary somatosensory cortex (SI) were observed ~20 to 50 ms after stimulus onset and were followed by additional left SI responses and bilateral responses in the secondary somatosensory cortex (SII). In addition, in the Continuous and Gap conditions, there was an activation of the precentral gyrus, the temporal aspects of which depended on the temporal relation of the administered stimuli, as follows. An ISI of 0 ms led to activation of the precentral gyrus shortly after the second stimulation, whereas an ISI of 50 ms led to activation of the precentral gyrus after the third stimulation. The current findings support results from previous studies on temporal activity patterns in SI and SII, verify the participation of the precentral gyrus during tactile motion perception and, in addition, reveal aspects of integration of sequential sensory stimulations over nonadjacent areas as well as temporal activity patterns in the postcentral and precentral gyri. PMID- 18512052 TI - Tactile acuity in experienced Tai Chi practitioners: evidence for use dependent plasticity as an effect of sensory-attentional training. AB - The scientific discovery of novel training paradigms has yielded better understanding of basic mechanisms underlying cortical plasticity, learning and development. This study is a first step in evaluating Tai Chi (TC), the Chinese slow-motion meditative exercise, as a training paradigm that, while not engaging in direct tactile stimulus training, elicits enhanced tactile acuity in long-term practitioners. The rationale for this study comes from the fact that, unlike previously studied direct-touch tactile training paradigms, TC practitioners focus specific mental attention on the body's extremities including the fingertips and hands as they perform their slow routine. To determine whether TC is associated with enhanced tactile acuity, experienced adult TC practitioners were recruited and compared to age-gender matched controls. A blinded assessor used a validated method (Van Boven et al. in Neurology 54(12): 2230-2236, 2000) to compare TC practitioners' and controls' ability to discriminate between two different orientations (parallel and horizontal) across different grating widths at the fingertip. Study results showed that TC practitioners' tactile spatial acuity was superior to that of the matched controls (P < 0.04). There was a trend showing TC may have an enhanced effect on older practitioners (P < 0.066), suggesting that TC may slow age related decline in this measure. To the best of our knowledge, this is the first study to evaluate a long-term attentional practice's effects on a perceptual measure. Longitudinal studies are needed to examine whether TC initiates or is merely correlated with perceptual changes and whether it elicits long-term plasticity in primary sensory cortical maps. Further studies should also assess whether related somatosensory attentional practices (such as Yoga, mindfulness meditation and Qigong) achieve similar effects. PMID- 18512053 TI - Medication errors made by health care professionals. Analysis of the Finnish Poison Information Centre data between 2000 and 2007. AB - INTRODUCTION: The purpose of this study was to analyse the extent, type and time trends of medication errors made by health care professionals leading to a call to the Finnish Poison Information Centre (PIC). METHODS: The PIC database consisting of all calls (277,300) received between 1 June 2000 and 31 May 2007 was analysed in terms of medication errors. RESULTS: Of 189,956 calls involving acute human poisonings, 1270 (0.7%) concerned medication errors (n = 1275), of which 779 (60.9%) involved administration of the wrong drug, 429 (33.6%) involved administration of the wrong dose and 70 (5.5%) involved erroneous route of administration. Incidents involving the elderly (80-89 years of age, n = 231; 18.2%) and children below 10 years (n = 136; 10.7%) were most likely to result in a call to the PIC about a medication error. In children, the most common error was wrong dose, while in adults, it was wrong drug. The number of medication errors was greatest during the summer months and in December. CONCLUSIONS: Medication errors seem to be different in children and the elderly. They are also more likely prone to occur during holiday seasons. PMID- 18512054 TI - Follow-up imaging after laparoscopic heminephrectomy in children. AB - BACKGROUND: The radiological findings after laparoscopic heminephrectomy in children are not widely reported. OBJECTIVE: To document the range of imaging appearances following laparoscopic heminephrectomy in children. MATERIALS AND METHODS: We reviewed all imaging and all radiology reports performed on children in our institution who had undergone laparoscopic heminephrectomy over a 6-year period. RESULTS: Between 2001 and 2007, 35 patients (24 girls, 11 boys) had laparoscopic heminephrectomy. The mean age at surgery was 3.5 years. The sites of surgery comprised 12 right upper, 5 right lower, 18 left upper and 3 left lower heminephrectomies. These numbers included three patients who had bilateral procedures. The most consistent postoperative finding was an avascular cyst related to the site of surgery, seen after 23 of 38 procedures. Of these 23 cysts, 13 were simple and 10 were septated and/or contained echogenic debris. CONCLUSION: A cyst related to the site of laparoscopic heminephrectomy is a frequent postoperative occurrence but is not widely recognized for being simply this. Simple, anechoic, and septated appearances are possible. This cyst may be confusing to the radiologist and the clinician if they are not aware of the history of laparoscopic heminephrectomy and the technique that has been used. Whilst the exact aetiology of this cystic lesion is uncertain, all the patients in our series had a heminephrectomy performed with the use of an Endoloop (Ethicon, Somerville, NJ). No patients have required intervention to date. PMID- 18512055 TI - Genetic modification of Lactobacillus plantarum by heterologous gene integration in a not functional region of the chromosome. AB - This report describes the vector-free engineering of Lactobacillus plantarum by chromosomal integration of an exogenous gene without inactivation of physiological traits. The integrative plasmid vector pP7B6 was derived from pGIP73 by replacing the cbh site, encoding the L. plantarum conjugated bile salt hydrolase, with the prophage fragment P7B6, from L. plantarum Lp80 (DSM 4229). Plasmid pP7B6NI was obtained by inserting the nisin immunity gene nisI of Lactococcus lactis subsp. lactis DSM 20729, preceded by the constitutive promoter P32 from the same strain, in a unique XbaI site of fragment P7B6 and was used to electrotransform L. plantarum Lp80. A food grade recombinant L. plantarum Lp80NI, with 480-fold higher immunity to nisin than the wild type, was derived by integration of pP7B6NI followed by the excision of pP7B6. Polymerase chain reaction tests demonstrated that the integration of nisI in the prophage region had occurred and that the erythromycin resistance marker from pP7B6 was lost. Fifteen among 31 L. plantarum strains tested hybridized with P7B6, indicating that the integration of pP7B6-derived vectors might occur in some other L. plantarum strains. This was experimentally confirmed by constructing the recombinant strain L. plantarum LZNI from the dairy isolate L. plantarum LZ (LMG 24600). PMID- 18512056 TI - Purification and characterization of Chromobacterium sp. DS-1 cholesterol oxidase with thermal, organic solvent, and detergent tolerance. AB - A new screening method for 6beta-hydroperoxycholest-4-en-3-one (HCEO)-forming cholesterol oxidase was devised in this study. As the result of the screening, a novel cholesterol oxidase producer (strain DS-1) was isolated and identified as Chromobacterium sp. Extracellular cholesterol oxidase of strain DS-1 was purified from the culture supernatant. The molecular mass of the purified enzyme was 58 kDa. This enzyme showed a visible adsorption spectrum having peaks at 355 and 450 nm, like a typical flavoprotein. The enzyme oxidized cholesterol to HCEO, with the consumption of 2 mol of O2 and the formation of 1 mol of H2O2 for every 1 mol of cholesterol oxidized. The enzyme oxidized 3beta-hydroxysteroids such as cholesterol, beta-cholestanol, and pregnenolone at high rates. The Km value for cholesterol was 26 microM. The enzyme was stable at pH 3 to 11 and most active at pH 7.0-7.5, showing optimal activity at pH 7.0 and 65 degrees C. The enzyme retained about 80% of its activity after incubation for 30 min at 85 degrees C. The thermal stability of the enzyme was the highest among the cholesterol oxidases tested. Moreover, the enzyme was more stable in the presence of various organic solvents and detergents than commercially available cholesterol oxidases. PMID- 18512057 TI - Purification, biochemical and molecular characterization of a metalloprotease from Pseudomonas aeruginosa MN7 grown on shrimp wastes. AB - A protease-producing bacterium was isolated and identified as Pseudomonas aeruginosa MN7. The strain was found to produce proteases when it was grown in media containing only shrimp waste powder (SWP), indicating that it can obtain its carbon, nitrogen, and salts requirements directly from shrimp waste. The use of 60 g/l SWP resulted in a high protease production. Elastase, the major protease produced by P. aeruginosa MN7, was purified from the culture supernatant to homogeneity using acetone precipitation, Sephadex G-75 gel filtration, and ultrafiltration using a 10-kDa cut-off membrane, with a 5.2-fold increase in specific activity and 38.4% recovery. The molecular weight of the purified elastase was estimated to be 34 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration. The optimum temperature and pH for protease activity were 60 degrees C and 8.0, respectively. The activity of the enzyme was totally lost in the presence of ethylene glycol tetraacetic acid, suggesting that the purified enzyme is a metalloprotease. The purified enzyme was highly stable in the presence of organic solvents, retaining 100% of its initial activity after 60 days of incubation at 30 degrees C in the presence of dimethyl sulfoxide and methanol. The lasB gene, encoding the MN7 elastase, was isolated and its DNA sequence was determined. PMID- 18512058 TI - Production of hydroxy-fatty acid derivatives from waste oil by Escherichia coli cells producing fungal cytochrome P450foxy. AB - Cytochrome P450foxy (P450foxy) is a fatty acid (FA) monooxygenase that is characterized by self-sufficient catalysis and high turnover numbers due to the fused structure of cytochrome P450 and its reductase. Here we found that resting recombinant Escherichia coli cells producing P450foxy converted saturated FA with a chain length of 7-16 carbon atoms to their omega-1 to omega-3 hydroxy derivatives. Most products were recovered from the culture supernatant. Decanoic acid was most efficiently converted to omega-1 to omega-3 hydroxy decanoic acids in the order of omega-1>omega-2>omega-3, with a total product yield of 47%. We also found that P450foxy was more active against physiological fatty acyl esters such as monopalmitoyl glycerol, monopalmitoyl phospholipid, and palmitoyl CoA than free palmitic acid. The bacteria producing P450foxy were applicable as biocatalysts in the production of omega-1 hydroxy palmitic acid from lard, vegetable, and soy sauce oil wastes from the food industry. PMID- 18512059 TI - The interaction of induction, repression and starvation in the regulation of extracellular proteases in Aspergillus nidulans: evidence for a role for CreA in the response to carbon starvation. AB - In Aspergillus nidulans, production of extracellular proteases in response to carbon starvation and to a lesser extent nitrogen starvation is controlled by XprG, a putative transcriptional activator. In this study the role of genes involved in carbon catabolite repression and the role of protein as an inducer of extracellular protease gene expression were examined. The addition of exogenous protein to the growth medium did not increase extracellular protease activity whether or not additional carbon or nitrogen sources were present indicating that induction does not play a major role in the regulation of extracellular proteases. Northern blot analysis confirmed that protein is not an inducer of the major A. nidulans protease, PrtA. Mutations in the creA, creB and creC genes increased extracellular protease levels in medium lacking a carbon source suggesting that they may have a role in the response to carbon starvation as well as carbon catabolite repression. Analysis of glkA4 frA2 and creADelta4 mutants showed that the loss of glucose signalling or the DNA-binding protein which mediates carbon catabolite repression did not abolish glucose repression but did increase extracellular protease activity. This increase was XprG-dependent indicating that the effect of these genes may be through modulation of XprG activity. PMID- 18512060 TI - Transanal one-stage endorectal pull-through for Hirschsprung's disease: a comparison with the staged procedures. AB - Transanal one-stage endorectal pull-through (TOSEPT) has been presented as the most recent progression in treatment of Hirschsprung's disease (HD), which may be able to replace various previous techniques. This prospective study was designed to compare the efficacy and probable complications of the commonly used open surgery with those of TOSEPT technique in management of HD. Forty-two children (35 boys and 7 girls) between 3 days and 12 years of age underwent surgical treatment for HD-all after being diagnosed by barium enema over an 18-month period. In a manner of systematic random selection, 21 patients were operated by TOSEPT and the other 21 by Swenson procedures. All patients were followed-up at least for 12 months after hospital discharge. The necessary data containing age, sex, length of the resected aganglionic segment, intraoperative details, duration of hospital stay and postoperative functional results or complications were collected during hospitalization and follow-up period. There was no significant difference in age at the first operation, sex distribution and length of resected bowel between the two groups. Rate of postoperative complications were significantly higher in Swenson procedure group (P < 0.01). Narcotics were needed in all patients of Swenson procedure group but only in two patients operated by TOSEPT also required laparotomy (P < 0.001). The hospitalization period of TOSEPT group was also less than that of Swenson group (P < 0.001). The total cost of treatment by Swenson procedure is considerably higher than by TOSEPT. TOSEPT can significantly diminish postoperative pain, surgical complications, hospital stay and cost burden caused by Hirschsprung's disease. Besides, this procedure is cosmetically preferable because no visible scar remains. PMID- 18512061 TI - The influence of trisomy 21 on the incidence and severity of congenital heart defects in patients with duodenal atresia. AB - Duodenal atresia is associated with a wide variety of congenital malformations. Trisomy 21 occurs in approximately one-thirds of infants with duodenal atresia. Congenital heart disease in patients with trisomy 21 and duodenal atresia is well known. However, the frequency and spectrum of congenital heart defects in infants with duodenal atresia and a normal karyotype has not been outlined in the literature. Therefore, we conducted a retrospective chart review to clarify our knowledge about this population. Retrospective review of the medical record was performed on patients with duodenal atresia/stenosis from January 1995 to September 2007. Demographic data included birth weight and gestational age. Variables of interest included cardiac defects and karyotype. Surgical repair for duodenal and cardiac malformations were reviewed. Ninety-four patients with duodenal atresia/stenosis were identified. Average gestational age was 36 weeks and birth weight was 2,536 g. Trisomy 21 was identified in 39 (41%) patients. Overall, 37 patients (39.3%) had a congenital heart defect. Defects were identified in 24 (61.5%) patients with trisomy 21, when compared to 13 (23.6%) patients with a normal karyotype. Of the patients with congenital heart defects and trisomy 21, 11 (28.2%) required operative repair compared to the 6 (10.9%) patients with a defect and normal karyotype. Therefore, in patients with duodenal atresia, the presence of trisomy 21 carries a relative risk of 2.61 for congenital heart defects, and relative risk of 2.59 for open heart surgery. In patients with duodenal atresia, the presence of trisomy 21 carries a 2.5-fold increased risk of cardiac defect and the same increased risk for repairing a cardiac defect. PMID- 18512062 TI - Evaluation of outcome of anorectal anomaly in childhood: the role of anorectal manometry and endosonography. AB - The aim of this study was to evaluate role of anorectal manometry (ARM) and anal endosonography (ES) in assessment of the internal anal sphincter (IAS) quality on continence outcome following repair of anorectal anomalies (ARA). We devised a scoring system to evaluate the quality of the IAS based on ARM and ES and correlated the scores with clinical outcome, using a modified Wingfield score (MWS) for faecal continence. We also assessed the implication of megarectum and neuropathy on faecal continence. Of 54 children studied, 34 had high ARA and 20 had low ARA. Children with high ARA had poor sphincters on ES and ARM, and also poor faecal continence compared to those with low ARA. The presence of megarectum and neuropathy was associated with uniformly poor outcome irrespective of the IAS quality. The correlations between MWS on one hand, and ES and ARM scores for IAS on the other hand were weak in the whole study group, ES r = 0.27, P < 0.04, and ARM r = 0.39, P < 0.004. However, the correlations were strong in those who had isolated ARA without megarectum or neuropathy, ES r = 0.51, P < 0.02 and ARM r = 0.55, P < 0.01, respectively. In conclusion, the ARM and ES are valuable in evaluation of continence outcome in children after surgery for ARA and those with good quality IAS had better faecal continence. The IAS is a vital component in functional outcome in absence of neuropathy and megarectum. PMID- 18512063 TI - Peptide YY induces enterocyte proliferation in a rat model with total enteral nutrition after distal bowel resection. AB - The main reason why enterocyte proliferative effects of peptide YY (PYY) have not been detected in rats undergoing massive small intestinal resection after feeding may have been the background activity of markedly increased endogenous PYY released from L cells in the distal gut in response to the intraluminal nutrients. The purpose of the present study was to evaluate the effects of PYY on enterocyte proliferation in a rat model of distal bowel resection (DBR) with total enteral nutrition (TEN). Male, adult Sprague-Dawley rats were assigned into three experimental groups: sham rats underwent bowel transection and reanastomosis, DBR rats underwent the resection of 40 cm distal small intestine and colon, and DBR-PYY rats underwent distal bowel resection as above, and were treated with PYY(1-36) from day 2 to day 14 postoperatively. During days 2-14 postoperatively, all animals received isocaloric TEN. At the endpoints, plasma PYY levels and parameters of enterocyte proliferation were determined. Compared with the sham group, DBR rats demonstrated a significant decrease in plasma PYY levels, and a significant increase in intestinal bowel and mucosal weight, mucosal DNA and protein content, villus height and crypt depth, and crypt cell proliferation index. Administration of PYY (DBR-PYY group) led to a significant increase in plasma PYY levels, intestinal bowel and mucosal weight, mucosal DNA and protein content, villus height and crypt depth, and crypt cell proliferation index in comparison with the DBR untreated group. We conclude that administration of PYY increases the plasma PYY levels, and PYY induces enterocyte proliferation with TEN after distal bowel resection. PMID- 18512064 TI - Prevalence of inflammatory bowel diseases in the Western Nations: high consumption of potatoes may be contributing. PMID- 18512065 TI - Patellar height before and after unicompartmental knee arthroplasty: association with early clinical outcome? AB - INTRODUCTION: There is paucity of information available concerning the role of patellar height in unicompartmental knee arthroplasty (UKA). The present study was conducted to determine the patellar height before and after UKA and to assess possible effects on the early clinical outcome. MATERIALS AND METHODS: We measured the patellar height before and after UKA in 83 consecutive knees using the Blackburne-Peel (BP) index and Insall-Salvati (IS) ratio and investigated the impact of the patellar height on the clinical outcome 2 years after surgery. RESULTS: BP values significantly decreased from 0.81 before surgery to 0.76 postoperatively (P < 0.001). IS ratios did not significantly decrease from 1.02 to 1.01 (P = 0.108). Lower preoperative BP values were negatively correlated with the postoperative knee extension (r = -0.357, P = 0.026), while higher preoperative BP values were negatively associated with the postoperative Knee Society Score (r = -0.302, P = 0.046). Lower preoperative IS values were negatively correlated with postoperative Knee Scores (r = -0.394, P = 0.019). CONCLUSION: After UKA, the patellar height decreased significantly according to the BP index, but not significantly according to the IS ratio. We found only weak and inconsistent correlations between the patellar height and clinical outcome parameters. Hence, based on the present results, the patellar height seems not to be a strict separate patient-selection criterion for UKA. PMID- 18512066 TI - Triplets on endometrial tuberculosis after two embryo transfer: a case report and the review of the literature. PMID- 18512067 TI - Hepatitis C virus budding at lipid droplet-associated ER membrane visualized by 3D electron microscopy. AB - The mechanisms underlying hepatitis C virus (HCV) morphogenesis remain elusive, but lipid droplets have recently been shown to be important organelles for virus production. We investigated the interaction between HCV-like particles and lipid droplets by three-dimensional reconstructions of serial ultrathin electron microscopy sections of cells producing the HCV core protein. The budding of HCV like particles was mostly initiated at membranes close to the lipid droplets rather than at membranes directly apposed to the lipid droplets. This may have important implications for our understanding of the complex relationship between HCV and lipids and may make easier to dissect out the HCV life cycle. PMID- 18512068 TI - Emotional exhaustion and mental health problems among employees doing "people work": the impact of job demands, job resources and family-to-work conflict. AB - OBJECTIVE: This study investigates the relationship between four job characteristics and family-to-work conflict on emotional exhaustion and mental health problems. METHODS: Multiple regression analyses were performed using data from 1,008 mental health care employees. Separate regression analyses were computed for high and low patient interaction jobs. RESULTS: Different job characteristics as well as family-to-work conflict were associated with emotional exhaustion and mental health problems in each job type. The relationship between family-to-work conflict and emotional exhaustion was mitigated by social support from colleagues for those who worked in low patient interaction jobs. CONCLUSION: In addition to general and specific stressors, it is worthwhile to include home related stressors that interfere with the work domain in stress research. PMID- 18512069 TI - Changes of noradrenaline-induced contractility and gene expression in aorta of rats acclimated to heat in two different modes. AB - We investigated changes in responsiveness to noradrenaline (NA) and gene expression in aorta, and plasma nitrite/nitrate (NOx) levels in heat-acclimated rats. Male rats were exposed for 10 consecutive days to either 32 degrees C in the second half of the dark phase (HI) or to 32 degrees C constantly (HC). Controls were kept at 24 degrees C. After heat acclimation, thoracic aorta was excised and blood was taken in the first (Dark1) and second (Dark2) halves of the dark phase, and in the light phase. Contractile response to NA in HI was significantly depressed in Dark1 and Dark2, whereas in HC the contraction response to NA was significantly attenuated in all phases. Endothelial NO synthase (eNOS) mRNA expression was significantly enhanced in HI in Dark2. Inducible NOS mRNA was undetectable. mRNA expression of period2 was not modified by heat acclimation. Plasma NOx concentrations in HI were significantly elevated in Dark1 and Dark2, while in HC, the level was significantly raised in all phases. Our results suggest that in rats, heat acclimation improves distensibility of thoracic aorta due to attenuated responsiveness to NA, which might be attributable in part to an upregulated eNOS expression and an increase in circulating NO. The phases of the day when such changes occurred appear to be closely related to the periods when the animals were previously subjected to heat. PMID- 18512070 TI - Adult living donor liver transplantation: body mass index and MELD score of recipients are independent risk factors for hospital mortality. AB - BACKGROUND AND AIMS: Adult living donor liver transplantation (LDLT) has been established as elective procedure or urgent procedure to save the life of patients with terminal liver diseases. The outcome of LDLT varies between transplant centers. Here, we aim to evaluate the outcome of LDLT in our center and to identify the risk factors that are associated with hospital mortality of recipients. PATIENTS AND METHODS: A cohort study with 32 consecutive cases of adult living donor liver transplantation was conducted in two cooperated medical centers. Perioperative data, incidence of postoperative complications, and hospital mortality were analyzed. RESULTS: No major surgical complications and no hospital mortality were observed in all 32 donors. All donors were discharged with normal liver function with median intensive care unit (ICU) stay of 1 day and median hospital stay of 10 days. All recipients had normal liver function in early posttransplant period. Eighty-one percent of the recipient survived with normal liver function for more than 1 year. The pretransplant ICU stay, renal failure, international normalized ratio (>1.8), and Model for End-stage Liver Disease (MELD) score (>20) were independent risk factors for hospital mortality of recipients. CONCLUSIONS: Adult living donor liver transplantation should be reserved to less "sick" patients in the era of organ allocation based on MELD score. PMID- 18512071 TI - Non-functioning, malignant pancreatic neuroendocrine tumour (PNET): a rare entity during pregnancy. AB - INTRODUCTION: Non-functioning pancreatic neuroendocrine tumours (PNET) are extremely rare tumours. To our knowledge, we are the first to report in the literature on two cases of PNET during pregnancy. PATIENTS: A 32- and a 35-year old primigravida, both in their second trimester of pregnancy, were referred to our clinic because of a suspicious mass in the pancreas. Both patients had an uneventful medical history and their laboratory test results, including pancreatic hormones, were unremarkable. Both underwent surgical exploration and resection of the pancreatic tumour. One patient with a tumour in the pancreatic tail underwent a distal pancreatectomy and splenectomy, whereas in the other case a pylorus-preserving pancreaticoduodenectomy was performed due to a tumour in the pancreatic head. RESULTS: One of the patients delivered a healthy infant, which is now at the age of 6 years in excellent health. The other patient is now in her third trimester of pregnancy without any further complications. Histopathological analysis of both tumours showed a strong expression of chromogranin and synaptophysin, thus both were classified as well-differentiated, non-functioning neuroendocrine carcinomas. CONCLUSIONS: Although PNETs are extremely rare, this entity can be confirmed as another differential diagnosis for suspicious pancreatic mass during pregnancy. In specialized centres, these tumours can be removed without any harmful consequences to the foetus. PMID- 18512072 TI - Evolutionary maintenance of oncogenesis. AB - BACKGROUND: Transformation of normal cells into cells with malignant phenotypes is often the result of loss of tumor suppressor gene (TSG) function after exposure to a carcinogen. CONCLUSIONS: We propose that TSGs susceptible to mutation and consequent loss of function are evolutionarily preserved in normal cell genomes so that the cells survive mutation-inducing insults and thereby evade apoptosis. While the mutations produced in TSGs confer cellular persistence and preclude apoptosis, oncogenesis is the untoward consequence. Proto-oncogenes might similarly be maintained and contain evolutionarily selected and fixed sequences susceptible to mutations (oncogene activation) that prevent cell death but ironically result in host death from malignancy. PMID- 18512073 TI - Gastrointestinal metastasis of lung cancer with special emphasis on a long-term survivor after operation. AB - PURPOSE: The aim of this study was to report the clinical characteristics and outcomes of gastrointestinal metastasis from lung cancer, with special emphasis on a long-term survivor after surgery. METHODS: We retrospectively reviewed all patients diagnosed with lung cancer between 2000 and 2007 at a tertiary referral center for cancer. RESULTS: Gastrointestinal metastases were detected in 10 (0.19%) of 5,239 lung cancer patients. Small bowel metastases occurred in one half of the patients, making it the most common metastatic site. One patient underwent emergency surgery because of an intestinal perforation. Although a perforation developed only in this patient, surgical intervention was required for five other patients to relieve intestinal obstruction or control bleeding. The prognosis was poor, with a median survival of 96.5 days after diagnosis. However, one patient remains alive >5 years post-operatively, without a recurrence, suggesting that surgical resection is an option for cure in properly selected patients. CONCLUSIONS: Physicians should be familiar with unique features of lung cancer with metastasis to the gastrointestinal tract so as to render early and optimal management. PMID- 18512074 TI - Clinical identification of colorectal cancer patients benefiting from adjuvant uracil-tegafur (UFT): a randomized controlled trial. AB - PURPOSE: A randomized controlled trial was conducted to determine whether pathologic necrosis in response to preoperative treatment with uracil tegafur(UFT) could be used to identify patients with colorectal cancer most likely to benefit from postoperative adjuvant therapy with the drug. PATIENTS AND METHODS: The 152 patients with colorectal cancer who received preoperative UFT at a dose of 600 mg/day for at least 10 days were classified into two groups according to the pathologic necrosis in resected tumor specimens: 90% or more necrosis (sensitive) versus less than 90% necrosis (insensitive). After excluding 13 ineligible patients, the remaining 139 were then randomly assigned to receive postoperative adjuvant UFT (400 mg/day) for 12 months or no treatment. RESULTS: Preoperative and postoperative UFT produced no serious toxicity in any of the patients. Among the 22 patients with sensitive tumors, overall survival was significantly better in the UFT group (n = 12) than in the control (n = 10) (100 vs. 70.0%; P = 0.023). Among the 117 patients with insensitive tumors, there was no significant difference between the two groups (n = 60, 68.1% vs. n = 57, 76.6%; P = 0.373). CONCLUSION: Our method involving neoadjuvant UFT can identify patients most likely to benefit from postoperative UFT, as well as those unlikely to benefit from such treatment. PMID- 18512075 TI - Chemotherapy of chronic indeterminate Chagas disease: a novel approach to treatment. AB - Treatment of Chagas disease is a controversial issue because the available drugs are highly toxic. Clomipramine is a tricyclic antidepressant drug that inhibits Trypanosoma cruzi's trypanothione reductase, provoking the death of the parasite and preventing the cardiac damage when used for the treatment of acutely infected mice. Here, we studied the effectiveness of clomipramine (5 mg/kg/day for one month) as chemotherapy for T. cruzi-infected mice in the chronic indeterminate stage of the infection. The animals were analyzed in the cardiac chronic phase. Survival of treated animals was 84% while for the untreated ones was 40%; most of the animals presented electrocardiographic abnormalities. Affinity and density of cardiac beta receptors from infected and treated mice were similar to those in the indeterminate phase, showing that clomipramine treatment stopped the increment of functional alterations provoked by the infection, while untreated mice presented affinity and density significantly diminished. Hearts from infected and untreated mice in the chronic stage presented mononuclear cells, necrosis and fiber dissolution while hearts from treated animals showed only isolated inflammatory infiltrates. Present results demonstrate that clomipramine used in the chronic indeterminate phase of the T. cruzi infection modified the natural evolution of the chagasic cardiopathy. PMID- 18512076 TI - Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and melting curve analysis. AB - A real-time fluorescence resonance energy transfer (FRET) PCR combined with melting curve analysis was developed for the detection of Opisthorchis viverrini in experimentally infected bithynid snails, its first intermediate hosts. The test is based on the fluorescence melting curve analysis of a hybrid between an amplicon from the pOV-A6-specific probe sequence, a 162-bp repeated sequence specific to O. viverrini and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single cercaria artificially introduced in a pool of 30 non-infected snails. The O. viverrini-infected snails were discriminated from non-infected snails and from genomic DNA of other parasite DNAs by their melting temperatures. Sensitivity and specificity of this method were both 100%. Melting curve analysis is a sensitive alternative for the specific detection of O. viverrini-infected snails; it is rapid, allows a high throughput, and can be done on small samples. The assay not only has a high potential for epidemiological surveys of O. viverrini-infected bithynid snails, but also for the detection of cercariae infestations of natural waterways when monitoring transmission sites. PMID- 18512077 TI - Outcomes of Schistosoma mansoni infection in outbred albino mice exposed to Larvin contaminant. AB - The present study tested the hypothesis that mice exposed to Schistosoma mansoni and treated with the insecticide Larvin have an increased risk of accelerated liver damage. To investigate this hypothesis, adverse effects resulting from treatment with Larvin were compared between S. mansoni-exposed and nonexposed outbred albino mice. The effects of concurrent treatment with Larvin on the progress and outcomes of S. mansoni infection were assessed via macroscopic and microscopic examination of liver and spleen, evaluation of several hematological, biochemical and hepatic enzymes parameters, and effect on worm burden. Oral administration of 1/5 and 1/10 LD(50) of Larvin to S. mansoni-exposed mice induced (1) hepatomegaly and splenomegaly; (2) prominent lymphocytic aggregation in liver replacing large areas of bridging necrosis; (3) increased serum level of bilirubin and alanine aminotransferase-aspartate aminotransferase enzymes; (4) decreased serum level of albumin and total proteins; and (5) decreased RBC, hemoglobin content, leukocyte, and lymphocyte counts. No significant effect on worm burden or oviposition was noted as a result of Larvin treatment compared to controls. All doses used in mice either for infection with S. mansoni cercariae or treatment with Larvin resulted in dose dependent alterations in hepatic functions of the tested mice. These alterations were most profound in mice exposed to S. mansoni and receiving Larvin treatment. The present findings support our hypothesis and show that concurrent S. mansoni infection with exposure to Larvin adversely affect liver functions and seriously alter hematological, biochemical, and hepatic enzymes parameters in outbred albino mice. These findings warrant further investigation and reinforce the need to minimize exposure to insecticide in both natural field settings and the broader environment. PMID- 18512078 TI - Novel microdeletion syndromes detected by chromosome microarrays. AB - Array comparative genomic hybridization (array CGH) has revolutionized the cytogenetic testing available for patients with learning disabilities who have "chromosomal" phenotypes with dysmorphic features and multiple anomalies. Screening large patient cohorts with mental retardation by array CGH has recently lead to the characterization of many novel microdeletion and microduplication syndromes, initially according to the shared cytogenetic aberrations, with secondary characterization of the corresponding phenotypes. This review provides a detailed clinical and molecular cytogenetic description of several of the most common of these aberrations. We have chosen to focus on patients in whom the cytogenetic abnormalities were principally described by array CGH, rather than by G-banded karyotyping or fluorescence in-situ hybridization. The syndromes that we have chosen include the 17q21.31 deletion and 17q21.31 duplication syndromes, 15q13.3 deletion syndrome, 16p11.2 deletion syndrome, 15q24 deletion syndrome, 1q41q42 deletion syndrome, 2p15p16.1 deletion syndrome and 9q22.3 deletion syndrome. In time, we hypothesize that at least some of these will become as clinically well characterized and recognizable to the clinician as the commoner microdeletion syndromes today. Although the full extent of the phenotypes is still evolving for many of these novel microdeletions, it is clear that array CGH has heralded an unparalleled era of discovery for clinical cytogenetics. PMID- 18512080 TI - Corrosion in bioprocessing applications. AB - Corrosion in bioprocessing applications is described for a 25-year-old bioprocessing pilot plant facility. Various available stainless steel alloys differ greatly in properties owing to the impact of specific alloying elements and their concentrations. The alloy property evaluated was corrosion resistance as a function of composition under typical bioprocessing conditions such as sterilization, fermentation, and cleaning. Several non-uniform forms of corrosion relevant to bioprocessing applications (e.g., pitting, crevice corrosion, intergranular attack) were investigated for their typical causes and effects, as well as alloy susceptibility. Next, the corrosion resistance of various alloys to specific bioprocessing-relevant sources of corrosion (e.g., medium components, acids/bases used for pH adjustment, organic acid by-products) was evaluated, along with the impact of temperature on corrosion progression. Best practices to minimize corrosion included considerations for fabrication (e.g., welding, heat treatments) and operational (e.g., sterilization, media component selection, cleaning) approaches. Assessments and repair strategies for observed corrosion events were developed and implemented, resulting in improved vessel and overall facility longevity. PMID- 18512081 TI - Diagnosing papillorenal syndrome: see the optic papilla. PMID- 18512082 TI - Nephronophthisis-like nephritis associated with fibrous dysplasia of bone. AB - Nephronophthisis is a chronic tubulointerstitial nephritis with autosomal recessive inheritance whose evolution to end-stage renal disease is insidious but constant. Fibrous dysplasia of bone is characterized by focal replacement of normal bone and marrow with abnormal bone and fibrous tissue. We report on a young boy initially diagnosed with fibrous dysplasia of bone, who underwent renal investigation because of treatment with pamidronate. He presented with mild proteinuria (albuminuria/creatininuria 19 mg/mmol) and decreased glomerular filtration rate (GFR) (79 ml/min per 1.73 m(2) body surface area) leading to kidney biopsy, which showed nephronophthisis-like lesions, but neither NPHP1 gene deletion nor UMOD (uromodulin) mutation were identified. No association between fibrous dysplasia of bone and nephronophthisis has yet been described. Nephronophthisis-like nephritis associated with fibrous dysplasia of bone might represent a possible new syndrome in the nephronophthisis and medullary cystic kidney disease complex. However, a fortuitous association between these two conditions is also possible. PMID- 18512083 TI - Responsiveness of the Brazilian-Portuguese version of the Oswestry Disability Index in subjects with low back pain. AB - When quality of life questionnaires are used as measures of treatment outcomes, it is essential to know how well these can respond to clinical changes. The objective of this study is to examine the responsiveness of the Brazilian Portuguese version of the Oswestry Disability Index (ODI-Brazil) in subjects with chronic low back pain submitted to a physical therapy program. Thirty subjects with chronic low back pain completed the ODI-Brazil questionnaire, along with an 11-point pain visual analogue scale (Pain VAS), and the Brazilian-Portuguese version of Roland-Morris disability questionnaire before and after the program. All patients also completed a global perception of change Likert scale in condition after the program. This scale was collapsed to produce a dichotomous variable outcome: improved and non-improved. Responsiveness was determined using effect size statistics and receiver operating characteristic curve (ROC curve), with best cut-point analysis. The best change score cut-off was identified when equally balanced sensitivity and specificity was found, as an expression of the minimum clinically important difference (MCID). After treatment, 19 patients considered themselves improved. Both the effect size (0.37) and the area under the ROC curve (0.73) for ODI-Brazil score in relation to global outcome after program indicated that the ODI-Brazil showed responsiveness. The ROC curve for ODI-Brazil was distributed at the upper corners of the diagonal line, indicating that the questionnaire presents discriminative ability. The best cut-off point for ODI-Brazil was approximately 4.45 points (63.2% sensitivity, 81.8% specificity). The Brazilian-Portuguese version of ODI has comparable responsiveness to other commonly used functional status measures and is appropriate for use in chronic low back pain patients receiving conservative care. PMID- 18512084 TI - Tissue identification with micro-magnetic resonance imaging in a caprine spinal fusion model. AB - Nonunion is a major complication of spinal interbody fusion. Currently X-ray and computed tomography (CT) are used for evaluating the spinal fusion process. However, both imaging modalities have limitations in judgment of the early stages of this fusion process, as they only visualize mineralized bone. Magnetic resonance imaging (MRI) could be of great value as it is able to discriminate between different types of tissue. A feasibility study was performed in nine animals from a goat spinal fusion study, to evaluate the detection capacity of different tissues with micro-MRI. In this study bioresorbable polylactic acid cages were used. Six- and 12-months follow-up specimens were scanned in a 6.3 T micro-MRI scanner. After scanning, the specimens were processed for histology. Different types of tissue as well as the degradable cage material were identified in the fusion zone and designated as regions of interest (ROIs). Subsequently, the location of these ROIs was determined on the corresponding micro-MRI image, and average signal intensities of every individual ROI were measured. An excellent match was seen between the histological sections and micro-MRI images. The micro-MRI images showed quantifiable differences in signal intensity between bone with adipose marrow, bone with hematopoietic marrow, fibrocartilage, fibrous tissue, and degradable implant material. In time the signal intensity of bone with adipose marrow, bone with hematopoietic red marrow, and of fibrous tissue remained relatively constant. On the other hand, the signal intensity of the degradable implant material and the fibrocartilage changed significantly in time, indicating change of structure and composition. In conclusion, in our model using bioresorbable cages the MRI provides us with detailed information about the early fusion process and may therefore, allow early diagnosis of non-union. PMID- 18512085 TI - Are early MRI findings correlated with long-lasting symptoms following whiplash injury? A prospective trial with 1-year follow-up. AB - Neck pain is the cardinal symptom following whiplash injuries. The trauma mechanism could theoretically lead to both soft tissue and bone injury that could be visualised by means of MRI. From previous quite small trials it seems that MRI does not demonstrate significant tissue damage. Large prospectively followed cohorts are needed to identify possible clinically relevant MRI findings. The objective of this trial was to evaluate (1) the predictive value of cervical MRI after whiplash injuries and (2) the value of repeating MRI examinations after 3 months including sequences with flexion and extension of the cervical spine. Participants were included after rear-end or frontal car collisions. Patients with fractures or dislocations diagnosed by standard procedures at the emergency unit were not included. MRI scans of the cervical spine were performed at baseline and repeated after 3 months. Clinical follow-ups were performed after 3 and 12 months. Outcome parameters were neck pain, headache, neck disability and working ability. A total of 178 participants had a cervical MRI scan on average 13 days after the injury. Traumatic findings were observed in seven participants. Signs of disc degeneration were common and most frequent at the C5-6 and C6-7 levels. Findings were not associated with outcome after 3 or 12 months. The population had no considerable neck trouble prior to the whiplash injury and the non-traumatic findings represent findings to be expected in the background population. Trauma-related MRI findings are rare in a whiplash population screened for serious injuries in the emergency unit and not related to a specific symptomatology. Also, pre-existing degeneration is not associated with prognosis. PMID- 18512079 TI - Genetic studies of African populations: an overview on disease susceptibility and response to vaccines and therapeutics. AB - Africa is the ultimate source of modern humans and as such harbors more genetic variation than any other continent. For this reason, studies of the patterns of genetic variation in African populations are crucial to understanding how genes affect phenotypic variation, including disease predisposition. In addition, the patterns of extant genetic variation in Africa are important for understanding how genetic variation affects infectious diseases that are a major problem in Africa, such as malaria, tuberculosis, schistosomiasis, and HIV/AIDS. Therefore, elucidating the role that genetic susceptibility to infectious diseases plays is critical to improving the health of people in Africa. It is also of note that recent and ongoing social and cultural changes in sub-Saharan Africa have increased the prevalence of non-communicable diseases that will also require genetic analyses to improve disease prevention and treatment. In this review we give special attention to many of the past and ongoing studies, emphasizing those in Sub-Saharan Africans that address the role of genetic variation in human disease. PMID- 18512086 TI - An insilico approach to high altitude pulmonary edema - Molecular modeling of human beta2 adrenergic receptor and its interaction with Salmeterol & Nifedipine. AB - Knowledge of the three-dimensional structures of protein targets from genomic data has the potential to accelerate researches pertaining to drug discovery. Human beta(2) adrenergic receptor is a G-protein-coupled receptor with seven transmembrane helices, and is important in pharmaceutical targeting on pulmonary and cardiovascular diseases. The human beta(2) adrenergic receptor has been found to play a very important role in the pathogenesis of high altitude pulmonary edema (HAPE). In the present study, a high quality of protein 3D structure has been predicted for the human beta(2) adrenergic receptor sequence with primary accession number P07550. Homologous template protein sequence with known 3D structure was identified and the template-query protein sequence validation was done by multiple sequence alignment method. The homology model was performed through Modeller and depended on the quality of the sequence alignment by BLAST, template structure and the consolidated result performed by Gene silico meta server. The statistical verification of the generated model was evaluated by PROCHECK which revealed that the structure modeled through Modeller to be of good quality with 84.1% of residues in the most favored region. Docking studies were carried out after modeling with two well known ligands namely Salmeterol and Nifedipine, and the fitness score revealed that Salmeterol has a higher fitness score than Nifedipine. Estimation of binding affinity by X-Score revealed that Salmeterol had -10.40 binding affinity while Nifedipine showed -9.62 binding affinity. From the present study, it can be concluded that the generated model of human beta(2) adrenergic receptor can be used for further studies related to this receptor and Salmeterol was found to have a high binding affinity with human beta(2) adrenergic receptor. PMID- 18512087 TI - Influence of azithromycin and clarithromycin on macrolide susceptibility of viridans streptococci from the oral cavity of healthy volunteers. AB - Oral viridans streptococci are a reservoir of resistance genes for pathogens. Through prolonged exposure, long-acting macrolides (e.g., azithromycin) may induce the resistance of the commensals to macrolides more frequently than macrolides with a shorter half-life (e.g., clarithromycin). In a prospective, randomized, evaluator-blinded trial in healthy volunteers receiving standard courses of either azithromycin (n = 20) or clarithromycin (n = 20), we compared the susceptibility of oral viridans streptococci to macrolides over a period of 12 weeks. There was a significant temporal increase in the numbers of resistant isolates in both groups (p < 0.0005 at week 1). The proportion of macrolide resistant isolates over time was significantly higher following azithromycin treatment (p = 0.0005), but returned to baseline values until week 12 in both groups. Temporal differential effects of azithromycin and clarithromycin on the induction of resistance were observed and need to be investigated regarding their effect on co-colonizing pathogens. PMID- 18512088 TI - Leishmaniases in Northern Greece: seroprevalence of the infection and incidence of the disease during the period 2001-2006. AB - Increasing risk factors are making leishmaniases a growing public health concern for many countries around the world. The aim of this study was to assess the seroprevalence of Leishmania infantum infection in the general population and in HIV infected subjects of Northern Greece, bordering the Mediterranean basin where leishmaniasis is endemic. The clinical cases of the disease during the last 6 years (2001-2006) are also presented. A low frequency of L. infantum antibodies was found by IFA and ELISA in 1,525 healthy individuals (2.8%), aged 18-80 years, living in the 16 prefectures of Northern Greece (Macedonia and Thrace regions), and in 167 HIV positive subjects (0.6%). Fifty-seven clinical cases were diagnosed in the same area and an approximate annual incidence of 0.34/100,000 was estimated. No endemic foci were identified and the cases of the disease were sporadic. Most presented with the visceral form (VL), few with the cutaneous, and one with VL-HIV co-infection. A significant shift in the age of people at risk was observed, with children less affected than adults (children/adults ratio: 0.36). No relevant data from previous studies are available to demonstrate a possible change of the infection in Northern Greece. The results of this study could be used as a reference for leishmaniasis surveillance in the area. PMID- 18512089 TI - Mycelial forms of Coccidioides spp. in the parasitic phase associated to pulmonary coccidioidomycosis with type 2 diabetes mellitus. AB - Pulmonary coccidioidomycosis shares characteristics with other pulmonary pathologies. In tissue, spherules containing endospores are markers of Coccidioides immitis and C. posadasii infection. Mycelial forms presenting without classical parasitic structures are often misdiagnosed. The study was performed at the National Institute of Respiratory Diseases (INER) of Mexico between September 1991 and June 2005 and analyzed the association between cases, controls, and risk factors, including co-morbidity. A case was defined as any patient who presented mycelial forms and a control as any patient who presented only spherules or no parasitic forms. All patients (n = 44) with pulmonary coccidioidomycosis were diagnosed by culture, histopathology, cytology, and immunology. Type 2 diabetic patients with pulmonary coccidioidomycosis were four times more likely than non-diabetics to develop parasitic mycelial forms (95% confidence interval [CI], 0.85-20.10; P < 0.01). We formulated a comprehensive definition based on the results as follows: patients with pulmonary coccidioidomycosis with an evolution longer than 8 months, cough, hemoptysis, radiological evidence of a cavitary lesion, and type 2 diabetes mellitus, develop parasitic mycelial forms of Coccidioides spp. Based on microscopic images of patient specimens, we propose incorporating mycelial forms into the parasitic phase of Coccidioides spp. in patients with type 2 diabetes mellitus and chronic and cavitary pulmonary coccidioidomycosis. PMID- 18512090 TI - Acute upper gastrointestinal hemorrhage: is a radiological interventional approach an alternative to emergency surgery? AB - The aim of our study was to discuss the option of endovascular treatment compared to surgery for patients with endoscopically unmanageable nonvariceal hemorrhage of the upper gastrointestinal tract. From 2000 to 2006, 23 patients (male, 15 male; female, 8; mean age, 69 years) who failed endoscopic therapy for upper gastrointestinal hemorrhage were retrospectively evaluated. Twelve patients were operated on (SG), whereas 11 patients had an endovascular intervention (IG). Technical and primary clinical success rates and complications rates were calculated. Clinical parameters and comorbidities were related to outcome. The surgical group suffered less frequently from pre-existing pulmonary diseases (SG, 17%; IG, 55%; p = 0.05) and had a higher incidence of shock requiring catecholamines (p < 0.01) or plasma expander therapy (p < 0.01). There was no significant difference in the incidence of recurrent bleeding episodes (SG, 17%; IG, 27%; p = 0.35) and mortality rates (SG, 17%; IG, 27%, p = 0.35). Deaths in the IG were due to recurrent bleeding. In patients with unsuccessful endoscopic control of nonvariceal bleeding of the upper GI tract, surgery remains a very effective treatment. However, in patients with a high surgical risk due to unknown bleeding sources and/or severe pre-existing diseases/comorbidities, endovascular therapy offers an excellent treatment option. These patients should then be operated on as early as possible to minimize the risk of recurrent bleeding episodes, which are associated with high morbidity and mortality. PMID- 18512091 TI - Dehydrin gene expression provides an indicator of low temperature and drought stress: transcriptome-based analysis of barley (Hordeum vulgare L.). AB - Low temperature and drought have major influences on plant growth and productivity. To identify barley genes involved in responses to these stresses and to specifically test the hypothesis that the dehydrin (Dhn) multigene family can serve as an indicator of the entire transcriptome response, we investigated the response of barley cv. Morex to: (1) gradual drought over 21 days and (2) low temperature including chilling, freeze-thaw cycles, and deacclimation over 33 days. We found 4,153 genes that responded to at least one component of these two stress regimes, about one fourth of all genes called "present" under any condition. About 44% (1,822 of 4,153) responded specifically to drought, whereas only 3.8% (158 of 4,153) were chilling specific and 2.8% (119 of 4,153) freeze thaw specific, with 34.1% responsive to freeze-thaw and drought. The intersection between chilling and drought (31.9%) was somewhat smaller than the intersection between freeze-thaw and drought, implying an element of osmotic stress response to freeze-thaw. About 82.4% of the responsive genes were similar to Arabidopsis genes. The expression of 13 barley Dhn genes mirrored the global clustering of all transcripts, with specific combinations of Dhn genes providing an excellent indicator of each stress response. Data from these studies provide a robust reference data set for abiotic stress. PMID- 18512092 TI - Fatty Acid Profile of Sunshine Bass: II. Profile Change Differs Among Fillet Lipid Classes. AB - Fatty acid (FA) profile of fish tissue mirrors dietary FA profile and changes in a time-dependent manner following a change in dietary FA composition. To determine whether FA profile change varies among lipid classes, we evaluated the FA composition of fillet cholesteryl esters (CE), phospholipids (PL), and triacylglycerols (TAG) of sunshine bass (SB, Morone chrysops x M. saxatilis) raised on feeds containing fish oil or 50:50 blend of fish oil and coconut, grapeseed, linseed, or poultry oil, with or without implementation of a finishing period (100% FO feed) prior to harvest. Each lipid class was associated with a generalized FA signature, irrespective of nutritional history: fillet PL was comprised largely of saturated FA (SFA), long-chain polyunsaturated FA (LC-PUFA), and total n-3 FA; fillet TAG was higher in MC-PUFA and total n-6 FA; and fillet CE was highest in monounsaturated FA (MUFA). Neutral lipids reflected dietary composition in a near-direct fashion; conversely, PL showed evidence of selectivity for MC- and LC-PUFA. Shorter-chain SFA were not strongly reflected within any lipid fraction, even when dietary availability was high, suggesting catabolism of these FA. FA metabolism in SB is apparently characterized by a division between saturated and unsaturated FA, whereby LC-PUFA are preferentially incorporated into tissues and SFA are preferentially oxidized for energy production. We demonstrated provision of SFA in grow-out feeds for SB, instead MC PUFA which compete for tissue deposition, meets energy demands and allows for maximum inclusion of LC-PUFA within fillet lipids. PMID- 18512093 TI - Prehospital cooling with hypothermia caps (PreCoCa): a feasibility study. AB - BACKGROUND: Animal studies suggest that the induction of therapeutic hypothermia in patients after cardiac arrest should be initiated as soon as possible after ROSC to achieve optimal neuroprotective benefit. A "gold standard" for the method of inducing hypothermia quickly and safely has not yet been established. In order to evaluate the feasibility of a hypothermia cap we conducted a study for the prehospital setting. METHODS AND RESULTS: The hypothermia cap was applied to 20 patients after out-of-hospital cardiac arrest with a median of 10 min after ROSC (25/75 IQR 8-15 min). The median time interval between initiation of cooling and hospital admission was 28 min (19-40 min). The median tympanic temperature before application of the hypothermia cap was 35.5 degrees C (34.8-36.3). Until hospital admission we observed a drop of tympanic temperature to a median of 34.4 degrees C (33.6-35.4). This difference was statistically significant (P < 0.001). We could not observe any side effects related to the hypothermia cap. 25 patients who had not received prehospital cooling procedures served as a control group. Temperature at hospital admission was 35.9 degrees C (35.3-36.4). This was statistically significant different compared to patients treated with the hypothermia cap (P < 0.001). CONCLUSIONS: In summary we demonstrated that the prehospital use of hypothermia caps is a safe and effective procedure to start therapeutic hypothermia after cardiac arrest. This approach is rapidly available, inexpensive, non-invasive, easy to learn and applicable in almost any situation. PMID- 18512094 TI - Real-time transesophageal three-dimensional echocardiography for guidance of percutaneous cardiac interventions: first experience. AB - Recently, a new generation of transesophageal echocardiography (TEE) probes with a novel matrix array technique was introduced, allowing three-dimensional (3D) presentation of cardiac structures in real-time. This article aims to describe our first experiences with this new technique in the guidance of percutaneous cardiac interventions in the catheter laboratory. We used a matrix array 3D TEE probe connected to a 3D-capable echocardiographic system. The 3D TEE system provides exact imaging of the pathomorphology of cardiac structures as well as intracardiac catheters and devices in real-time. We applied this innovative technique to monitor percutaneous cardiac interventions in the catheter laboratory, such as atrial septal defect (ASD) or patent foramen ovale (PFO) closures, revalving procedures such as percutaneous transvenous mitral valve annuloplasty (PTMA), aortic valve replacements, and electrophysiological procedures. Our findings demonstrate that real-time 3D TEE provides a novel imaging technique to guide interventions in the catheter laboratory, providing fast and complete information about the underlying pathomorphology, improving spatial orientation, and additionally monitoring online the procedure without loss of image quality. These benefits may accelerate the learning curve and improve confidence of the interventional cardiologist in order to increase safety, accuracy, and efficacy of interventional cardiac procedures. PMID- 18512095 TI - Inhibiting matrix metalloproteinase-2 reduces protein release into coronary effluent from isolated rat hearts during ischemia-reperfusion. AB - BACKGROUND: Previous studies have shown that the disruption of the coronary endothelium and the increase in its permeability during ischemia-reperfusion (I/R), are linked to matrix metalloproteinase-2 (MMP-2) activity. Studies from our group have shown that during I/R, activity of MMP-2 in the coronary effluent increases and this increase is associated with cardiac dysfunction, which in turn, can be prevented by MMP inhibitors. Therefore, we hypothesize that inhibiting MMPs reduces the MMP-2 dependent disruption of the coronary endothelium and subsequent protein release during I/R. METHODS: Isolated rat hearts were perfused in the Langendorff mode at a constant pressure and subjected to 15, 20 or 30 min no-flow ischemia followed by 30 min of reperfusion. The MMP inhibitors, o-phenanthroline (Phen, 100 microM) or doxycycline (Doxy, 30 microM) an inhibitors of MMPs, were added to the perfusion solution 10 min before ischemia and for the first 10 min of reperfusion. The coronary effluents were collected during perfusion for protein analysis. Creatine kinase was measured as an index of cellular damage. Endothelial integrity was assessed by measuring coronary flow and by measuring the levels of serotransferrin and interstitial albumin in the coronary effluent. Additionally, damage to the endothelium was assessed histologically by light microscopy analysis of the cellular structure of the myocardium. MMP-2 activity was measured by zymography in hearts subjected to 15, 20 and 30 min of ischemia without reperfusion. RESULTS: MMP-2 activity was increased in heart tissue at the end of ischemia and was correlated with duration of ischemia. The post-ischemia decrease in coronary flow, and the increase in the release of serotransferrin and albumin were attenuated by Phen. Edema (another indirect marker of endothelial damage) was observed in I/R heart and the edema was abolished in I/R heart treated with MMP inhibitors. CONCLUSION: MMP inhibition not only reduces cardiac mechanical dysfunction but also reduces endothelial damage resulting from cardiac I/R injury. PMID- 18512096 TI - Hsp70 inhibits aminoglycoside-induced hair cell death and is necessary for the protective effect of heat shock. AB - Sensory hair cells of the inner ear are sensitive to death from aging, noise trauma, and ototoxic drugs. Ototoxic drugs include the aminoglycoside antibiotics and the antineoplastic agent cisplatin. Exposure to aminoglycosides results in hair cell death that is mediated by specific apoptotic proteins, including c-Jun N-terminal kinase (JNK) and caspases. Induction of heat shock proteins (Hsps) is a highly conserved stress response that can inhibit JNK- and caspase-dependent apoptosis in a variety of systems. We have previously shown that heat shock results in a robust upregulation of Hsps in the hair cells of the adult mouse utricle in vitro. In addition, heat shock results in significant inhibition of both cisplatin- and aminoglycoside-induced hair cell death. In our system, Hsp70 is the most strongly induced Hsp, which is upregulated over 250-fold at the level of mRNA 2 h after heat shock. Therefore, we have begun to examine the role of Hsp70 in mediating the protective effect of heat shock. To determine whether Hsp70 is necessary for the protective effect of heat shock against aminoglycoside induced hair cell death, we utilized utricles from Hsp70.1/3 (-/-) mice. While heat shock inhibited gentamicin-induced hair cell death in wild-type utricles, utricles from Hsp70.1/3 (-/-) mice were not protected. In addition, we have examined the role of the major heat shock transcription factor, Hsf1, in mediating the protective effect of heat shock. Utricles from Hsf1 (-/-) mice and wild-type littermates were exposed to heat shock followed by gentamicin. The protective effect of heat shock on aminoglycoside-induced hair cell death was only observed in wild-type mice and not in Hsf1 (-/-) mice. To determine whether Hsp70 is sufficient to protect hair cells, we have utilized transgenic mice that constitutively overexpress Hsp70. Utricles from Hsp70-overexpressing mice and wild-type littermates were cultured in the presence of varying neomycin concentrations for 24 h. The Hsp70-overexpressing utricles were significantly protected against neomycin-induced hair cell death at moderate to high doses of neomycin. This protective effect was achieved without a heat shock. Taken together, these data indicate that Hsp70 and Hsf1 are each necessary for the protective effect of heat shock against aminoglycoside-induced death. Furthermore, overexpression of Hsp70 alone significantly inhibits aminoglycoside induced hair cell death. PMID- 18512097 TI - Recording and reenactment of collaborative diagnosis sessions using DICOM. AB - This paper presents a radiological collaborative tool capable of direct manipulation of Digital Imaging and Communications in Medicine (DICOM) images on both sides, and also recording and reenacting of a recorded session. A special collaborative application protocol formerly developed was extended and used as basis for the development of collaborative session recording and playback processes. The protocol is used today for real-time radiological meetings through the Internet. This new standard for collaborative sessions makes possible other uses for the protocol, such as asynchronous collaborative sessions, decision regulation, auditing, and educational applications. Experimental results are given which compare this protocol with other popular collaborative approaches. Comparison of these results shows that the proposed protocol performs much better than other approaches when run under controlled conditions. PMID- 18512098 TI - Preoperative and postoperative quality of life in patients with familial adenomatous polyposis. AB - PURPOSE: This study was designed to prospectively examine functional outcome, quality of life, and patients' personal experiences and adjustment to functional changes during the first year after prophylactic surgery. METHODS: Twenty-one consecutive patients with familial adenomatous polyposis were examined before proctocolectomy (T0), on ileostomy reversal (T1), and 6 (T2) and 12 months (T3) after surgery by means of standardized questionnaires and interviews. RESULTS: Average physical and mental health declined profoundly after proctocolectomy, followed by a steady improvement after 6 and 12 months. The majority of patients reported the ileostomy period as particularly distressing. After one year, 75 percent of patients reported complete recovery in terms of physical, emotional, and social functioning, whereas one-quarter of patients did not regain their former level of functioning. Despite substantial improvement in pouch functions, functional impairment persists because of frequent bowel movements, resulting mainly in restricted social activities. Ten percent of patients reported impaired sex life, irrespective of gender. CONCLUSIONS: The majority of patients with familial adenomatous polyposis were found to adjust favorably to functional impairment while maintaining satisfactory quality of life. Complementing standardized quality of life measures by patients' personal experiences may help to identify vulnerable patients in need of psychosocial support. PMID- 18512099 TI - Tracking outcomes of anorectal surgery: the need for a disease-specific quality assessment tool. AB - PURPOSE: We sought to determine the nature and timing of complications after common anorectal operations by using a prospective quality tracking tool. METHODS: A prospectively maintained quality database was queried to identify patients who underwent pilonidal sinus excision, hemorrhoidectomy, sphincterotomy, abscess drainage, or fistulotomy during an 11-year interval. All hospital complications were recorded by a single nurse practitioner and verified jointly by the surgical team. Any posthospital complications were registered at the first postoperative visit. RESULTS: A total of 969 patients underwent one of the five index anorectal procedures during the study period. Forty-nine complications occurred in 38 patients (3.9 percent). The majority of complications were minor (40/49; 82 percent) and were primarily urinary retention, minor bleeding, and wound infection. Twenty-five of the 40 minor complications (62 percent) were identified only after hospital discharge in the outpatient setting. Eight of the nine major complications occurred in patients already hospitalized for major concomitant illnesses and were unrelated to the anorectal surgery. The remaining patient had a postoperative deep vein thrombosis. CONCLUSIONS: Complications after anorectal procedures are infrequent, typically minor, and occur after hospital discharge. Major complications reflect concomitant illness, not surgical quality. Meaningful outcome measures are needed to assess the quality of anorectal surgery. PMID- 18512101 TI - Complication rates after Hartmann's reversal: open vs. laparoscopic approach. AB - PURPOSE: This study was performed to compare open Hartmann's reversal to laparoscopic Hartmann's reversal with regard to complication, readmission, and reoperation rates. METHODS: Data of patients who underwent open Hartmann's reversal or laparoscopic Hartmann's reversal between 1998 and 2004 at two institutions were collected. End points were complications in the hospital or after discharge, readmission to the hospital, and reoperation within 6 months after initial surgery. RESULTS: Sixty-one open Hartmann's reversal and 61 laparoscopic Hartmann's reversal patients were well matched except for American Society of Anesthesiology grade (1.9 vs. 1.6; P = 0.008), timing of Hartmann's procedure (14 vs. 6 months; P = 0.001), operation time (210 vs. 154 minutes; P = 0.001), and estimated blood loss (363 vs. 254 ml; P = 0.01). Thirty-day complication rates did not differ (18 vs. 13 percent). At 6 month follow-up, open Hartmann's reversal patients had increased complication (16.4 vs. 3.3 percent; P = 0.015) and reoperation (13.1 vs. 3.3 percent; P = 0.048) rates but the same readmission rates (16.4 percent). CONCLUSIONS: Compared with open Hartmann's reversal, 6 month complication and reoperation rates were lower in laparoscopic Hartmann's reversal patients. Most of the six-month complications and reoperations in open Hartmann's reversal were abdominal wall-related. Readmission rates were similar, but reasons for readmission were surgical in open Hartmann's reversal and medical in laparoscopic Hartmann's reversal. PMID- 18512102 TI - Methods of detection of first recurrence in patients with stage I/II primary cutaneous melanoma after sentinel lymph node biopsy. AB - BACKGROUND: An understanding of the methods of detection of recurrent melanoma after sentinel lymph node biopsy (SLNB) is essential for the coordination of a rational plan of follow-up. METHODS: Clinical stage I/II melanoma patients who underwent SLNB from 1991 to 2004 were identified from a prospectively maintained single-institution database. Detection of recurrence by self (awareness of symptoms or abnormal physical findings) or physician (discovered on routine physical or scheduled test) and timing of clinic visit were recorded. Postoperative follow-up included physical exam every 3-4 months for the first year, every 3-6 months for the second year, and every 6-12 months thereafter. Serum lactate dehydrogenase (LDH) and chest X-ray (CXR) were obtained annually. Computed tomography (CT) and positron emission tomography (PET) were performed selectively. RESULTS: Of 1062 patients who underwent SLNB, 203 (19%) experienced 230 initial sites of recurrence; 198 patients were evaluable for follow-up. Median follow-up after first recurrence was 17 months. Symptoms and self-detected physical findings were present in 109 patients (55%); 85 patients (78%) were seen earlier than their scheduled visit. Self-detected physical findings identified in transit (n = 26; 24%) and nodal (n = 25; 23%) disease. Physician detection occurred in 89 patients (45%), nearly half by a scheduled radiographic test (CXR, 16%; CT, 29%; PET, 1%). The method of detection significantly predicted post recurrence survival (p < 0.05). CONCLUSION: More than half of melanoma recurrences are self-detected; these patients have the most favorable post recurrence survival rates because of the type of recurrence detected. The mode of detection is a significant predictor of post-recurrence survival. This supports an aggressive program of patient education in self-examination after SLNB for melanoma. PMID- 18512103 TI - Videothoracoscopic resection of encapsulated thymic carcinoma: retrospective comparison of the results between thoracoscopy and open methods. AB - BACKGROUND: Although videothoracoscopic (VTS) resection of thymoma has been reported to be a less invasive technique than open sternotomy, the usefulness of this method in the treatment of encapsulated thymic carcinoma has not yet been evaluated. We retrospectively compared the VTS and open methods (median sternotomy) to investigate whether VTS resection could be performed as successfully as open surgery to treat resectable thymic carcinoma. METHODS: Between November 2002 and March 2007 a retrospective review was made of eight patients (four women and four men) with Masaoka stage I and II encapsulated thymic tumor. Four patients (the VTS group) underwent tumor resection by means of a three-port endoscopic technique. The other four patients (the open group) underwent tumor excision via a standard sternotomy approach. The resected thymic carcinoma tissues were all confirmed by histopathological examination. RESULTS: No patient died nor did any major morbidity or recurrence occur during the mean follow-up period of 3.76 +/- 1.43 years. The open group sustained more blood loss (246.3 ml more) and pleural drainage time (5.7 days more), and were hospitalized for a longer period (12.5 days more). However in the open group the tumor size was larger (38.6 cm(3 )more ) and the mean follow-up time was longer (1.4 years more). CONCLUSION: These results have encouraged us to treat more patients with encapsulated thymic carcinoma by means of VTS resection. PMID- 18512105 TI - Surgical management of carotid body tumors. AB - BACKGROUND: Carotid body tumors (CBT) should be considered when evaluating every lateral neck mass. METHODS: A retrospective study was conducted of 52 patients with 57 CBT. The surgical approach and complications were reviewed. All patients were operated on without preoperative embolization. RESULTS: Multifocal paraganglioma (PG) were detected in six cases. A succinate dehydrogenase subunit D (SDHD) mutation was discovered in four patients. Vascular peroperative complication occurred in one case. Vascular reconstruction was decided peroperatively in five cases (8.8%). Vascular reconstruction was 0% for Shamblin 1 or 2 tumors, but 28.5% for Shamblin 3. A postoperative nerve paresis was reported in 24 patients (42.1%) and vagal nerve paralysis persisted in four cases (7.01%). The rate of serious complications, e.g., permanent nerve palsy, preoperative and postoperative complications, was 14.03%; it was 2.3% for Shamblin 1 or 2 tumors and 35.7% for Shamblin 3. One patient had malignant PG with node metastasis and was not referred for radiotherapy. No recurrence or metastasis was reported after 6-year follow-up. CONCLUSION: Early surgical treatment is recommended in almost all patients after preoperative evaluation and detection of multifocal tumors. Surgical excision of small tumors was safe and without complication, but resection of Shamblin 3 tumors can be challenging. Routine preoperative embolization of carotid body paragangliomas is not required. PMID- 18512106 TI - Synchrony and asynchrony in a fully stochastic neural network. AB - We describe and analyze a model for a stochastic pulse-coupled neural network, in which the randomness in the model corresponds to synaptic failure and random external input. We show that the network can exhibit both synchronous and asynchronous behavior, and surprisingly, that there exists a range of parameters for which the network switches spontaneously between synchrony and asynchrony. We analyze the associated mean-field model and show that the switching parameter regime corresponds to a bistability in the mean field, and that the switches themselves correspond to rare events in the stochastic system. PMID- 18512107 TI - A geometric comparison of single chain multi-state models of ion channel gating. AB - Multi-state models of ion channel gating have been used extensively, but choosing optimally small yet sufficiently complex models to describe particular experimental data remains a difficult task. In order to provide some insight into appropriate model selection, this paper presents some basic results about the behavior of solutions of multi-state models, particularly those arranged in a chain formation. Some properties of the eigenvalues and eigenvectors of constant rate multi-state models are presented. A geometric description of a three-state chain is given and, in particular, differences between a chain equivalent to an Hodgkin-Huxley model and a chain with identical rates are analyzed. One distinguishing feature between these two types of systems is that decay from the open state in the Hodgkin-Huxley model is dominated by the most negative eigenvalue while the identical rate chain displays a mix of modes over all eigenvalues. PMID- 18512108 TI - The effects of scolicidal agent propolis on liver and biliary tree. AB - BACKGROUND: This study was designed to examine the effects of propolis on the liver and biliary system when used as a scolicidal agent. MATERIALS AND METHODS: Thirty Wistar-Albino rats were divided into two groups. Propolis and 0.9% saline (NaCl) were injected into the biliary tract of the rats. Three rats from control group and four rats from propolis group died within 5 days after the procedure. Blood samples of remaining 23 rats were obtained 1 week after and at the end of the experimental study for liver function tests. Six months after the procedure, retrograde and magnetic resonance cholangiography were performed and liver, common bile duct, and duodenum were excised en bloc for histopathological examination. RESULTS: Liver function tests were slightly elevated 1 week after the procedure and were found to be normal at the end of the sixth month in both groups. No stricture in the biliary tree was found on the retrograde and magnetic resonance cholangiograms. The tissue samples of the propolis group showed no histomorphological difference from the control group. CONCLUSIONS: Propolis may be used as a scolicidal agent even in the case of cystobiliary communication with no side effects on liver and biliary tree. PMID- 18512109 TI - CT findings in diffuse rectosigmoid cavernous heamangioma. A case report. PMID- 18512110 TI - Gastroesophageal reflux disease in morbid obesity: the effect of Roux-en-Y gastric bypass. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) is a common condition in obesity. The impact of Roux-en-Y gastric bypass (RYGBP) on GERD is poorly known. We studied the effect of the RYGBP on GERD in patients with morbid obesity (MO). METHODS: Twenty consecutive patients with MO (BMI > 40 kg/m(2)) were studied before and 6 months after RYGBP. GERD symptoms were evaluated with Carlsson-Dent questionnaire (CDQ). All the patients underwent esophageal manometry and ambulatory 24-h pH-metry. Chi-square test was used to compare categorical variables, and Wilcoxon test was used for numerical variables. A p value under 0.05 was considered significant. RESULTS: There were 16 women (80%) and 4 men (20%) with mean age 38.9 +/- 6.9 years included in this study. BMI was 48.5 +/- 6.2 kg/m(2) and 33.2 +/- 4.5 kg/m(2) before and after RYGBP, respectively. Mean weight reduction was 42.5 +/- 9.7 kg (p < 0.001). Reflux symptoms measured by CDQ and esophageal acid exposure improved significantly after RYGBP. The percentage of time of pH < 4 was 10.7 +/- 6.7 before and 1.6 +/- 1.2 after the surgical procedure (p < 0.001). LES basal pressure before and after the RYGBP was 18 +/- 11 and 20.1 +/- 5.6 mmHg (p = 0.372), and the esophageal body amplitude was 104.2 +/- 47.2 and 75.1 +/- 36.2 mmHg, respectively (p = 0.005). CONCLUSION: RYGBP improves GERD symptoms and reduces esophageal acid exposure in patients with MO. PMID- 18512111 TI - Engineering of functional cartilage tissue using stem cells from synovial lining: a preliminary study. AB - Stem cells derived from synovial lining-synovial lining--derived stem cells or SDSCs--are a promising cell source for cartilage tissue engineering. We hypothesized that negatively selected SDSCs would form cartilage constructs and conventionally passaged SDSCs would be contaminated with macrophages, inhibiting SDSC-based chondrogenesis. We mixed SDSCs with fibrin gel and seeded the cells into polyglycolic acid scaffolds. After 3 days of incubation with a proliferative growth factor cocktail (containing transforming growth factor beta1 [TGF-beta1], insulin-like growth factor I [IGF-I], and basic fibroblast growth factor [FGF 2]), the cell-fibrin-polyglycolic acid constructs were transferred into rotating bioreactor systems and cultured with a chondrogenic growth factor cocktail (TGF beta1/IGF-I) for up to 4 weeks. Tissue constructs based on negatively selected SDSCs had cartilaginous characteristics; were rich in glycosaminoglycans and collagen II; exhibited high expression of mRNA and protein for collagen II, aggrecan, and Sox 9; exhibited a negligible level of mRNA and protein for collagens I and X; and had an equilibrium modulus in the range of values measured for native human cartilage. Conventional passage yielded SDSCs with contaminating macrophages, which adversely affected the quality of tissue-engineered cartilage. We thus propose functional cartilage constructs could be engineered in vitro through the use of negatively isolated SDSCs. PMID- 18512112 TI - Mechanoactive scaffold induces tendon remodeling and expression of fibrocartilage markers. AB - Biological fixation of soft tissue-based grafts for anterior cruciate ligament (ACL) reconstruction poses a major clinical challenge. The ACL integrates with subchondral bone through a fibrocartilage enthesis, which serves to minimize stress concentrations and enables load transfer between two distinct tissue types. Functional integration thus requires the reestablishment of this fibrocartilage interface on reconstructed ACL grafts. We designed and characterized a novel mechanoactive scaffold based on a composite of poly-alpha hydroxyester nanofibers and sintered microspheres; we then used the scaffold to test the hypothesis that scaffold-induced compression of tendon grafts would result in matrix remodeling and the expression of fibrocartilage interface related markers. Histology coupled with confocal microscopy and biochemical assays were used to evaluate the effects of scaffold-induced compression on tendon matrix collagen distribution, cellularity, proteoglycan content, and gene expression over a 2-week period. Scaffold contraction resulted in over 15% compression of the patellar tendon graft and upregulated the expression of fibrocartilage-related markers such as Type II collagen, aggrecan, and transforming growth factor-beta3 (TGF-beta3). Additionally, proteoglycan content was higher in the compressed tendon group after 1 day. The data suggest the potential of a mechanoactive scaffold to promote the formation of an anatomic fibrocartilage enthesis on tendon-based ACL reconstruction grafts. PMID- 18512113 TI - Radioprotection of tendon tissue via crosslinking and free radical scavenging. AB - Ionizing radiation could supplement tissue bank screening to further reduce the probability of diseases transmitted by allografts if denaturation effects can be minimized. It is important, however, such sterilization procedures be nondetrimental to tissues. We compared crosslinking and free radical scavenging potential methods to accomplish this task in tendon tissue. In addition, two forms of ionizing irradiation, gamma and electron beam (e-beam), were also compared. Crosslinkers included 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and glucose, which were used to add exogenous crosslinks to collagen. Free radical scavengers included mannitol, ascorbate, and riboflavin. Radioprotective effects were assessed through tensile testing and collagenase resistance testing after irradiation at 25 kGy and 50 kGy. Gamma and e-beam irradiation produced similar degenerative effects. Crosslinkers had the highest strength at 50 kGy, EDC treated tendons had 54% and 49% higher strength than untreated, for gamma and e-beam irradiation respectively. Free radical scavengers showed protective effects up to 25 kGy, especially for ascorbate and riboflavin. Crosslinked samples had higher resistance to collagenase and over a wider dose range than scavenger-treated. Of the options studied, the data suggest EDC precrosslinking or glucose treatment provides the best maintenance of native tendon properties after exposure to ionizing irradiation. PMID- 18512115 TI - Interstitial lung disease and Sjogren's syndrome in primary biliary cirrhosis: a causal or casual association? AB - Interstitial lung disease (ILD) has been reported in patients with primary biliary cirrhosis (PBC); however, its frequency and pathogenesis are still poorly documented. Sjogren's syndrome (SS) is fairly common among patients with PBC, but the relationship between SS and PBC also remains controversial. To determine whether ILD and SS in PBC is a causal or casual association, whether SS accompanying PBC, could be considered secondary to or associated with PBC. One hundred and nine consecutive PBC cases were analyzed, and the differences of clinical features, histological stages, and serum autoantibodies between the PBC patients with and without SS were compared. There were 46 PBC patients with SS and 63 without SS, and 11 patients met the criteria of ILD. SS is associated with PBC in the form of secondary SS. The frequency of ILD in PBC patients with SS was 21.7% while only 1.6% in PBC patients without SS (P<0.0001). ILD in PBC was related to SS, with Spearman's rank coefficient of 0.330 (P=0.000). The association of SS with PBC, significantly higher in patients with than without ILD, which supports the hypothesis that ILD and SS in PBC, may be a causal, not casual, association. PMID- 18512114 TI - Relationship between bone quantitative ultrasound and mortality: a prospective study. AB - In a cohort of 5,201 women [72.3 +/- 5.3 years] from 58 primary care centers in Spain, followed for three years, no relationship between heel QUS parameters and overall mortality was found. However, a significant relationship between a low speed of sound (SOS) and vascular mortality was observed. INTRODUCTION: An inverse relationship between mortality and bone mineral density measured by dual energy absorption densitometry or quantitative bone ultrasound (QUS) has been described. The aim of the present study was to test this relationship in the ECOSAP cohort, a 3-year prospective study designed to assess the ability of heel QUS and clinical risk factors to predict non-vertebral fracture risk in women over 64. METHODS: A cohort of 5,201 women [72.3 +/- 5.3 years] was studied. QUS was assessed with the Sahara(R) bone sonometer. Women attended follow-up visits every 6 months. Physicians recorded if the patient died and cause of death. Hazard rates (HR) of all-cause and vascular mortality per one standard deviation reduction in QUS parameters were determined. RESULTS: One hundred (1.9%) women died during a median of 36.1 months follow-up, for a total of 14,999 patient years, 42 because of vascular events (both cardiovascular and cerebrovascular). After adjusting for age, none of the QUS variables showed statistically significant differences between the patients who died and the survivors. In the final multivariate model, adjusted for age, current thyroxine and hypoglycaemic drug use, chronic obstructive pulmonary disease and decreased visual acuity, SOS was marginally non-significant: (HR: 1.19; 0.97-1.45). However, each 1 SD reduction in SOS was associated with a 39% increase in vascular mortality (HR: 1.39; 1.15-1.66). CONCLUSIONS: In our cohort, SOS was related with vascular mortality, but not overall mortality. PMID- 18512116 TI - Relapse of chronic myeloid leukemia-chronic phase 14 years after allogeneic hematopoietic stem cell transplantation. PMID- 18512117 TI - Expression of green fluorescent protein under the regulation of human locus control region elements HS2 and HS3 in transgenic mice. AB - Human beta-globin locus control region (LCR) is composed of five DNase I hypersensitive sites (HSs). We previously demonstrated that when HS2 and HS3 were constructed together in one vector and integrated into one position in transgenic mice, the two cis-elements showed a marked synergy in regulating the spatial and temporal expression of beta-globin transgene. This study is to investigate whether these two elements still show a synergy or superposition when they are not located in one construct. HS2/GFP and HS3/GFP transgenic mice, with the transgene encoding green fluorescent protein (GFP) driven by beta-globin promoter and under the control of HS2 (HS2/GFP mice) and HS3 (HS3/GFP mice), were mated to generate double-heterozygotes (HS2/GFP + HS3/GFP mice). The expression and integration status of GFP genes in transgenic mice were analyzed. Our data showed that there was no difference in the percentages of GFP positive cells in peripheral blood between double-heterozygotes and their corresponding founders (HS2/GFP or HS3/GFP). Interestingly, it was observed that exogenous GFP genes were highly expressed in another transgenic mouse pedigree with HS2 element integrated twice into the same chromosome at two different loci. However, when one site was lost in some offspring because of recombination, the percentages of GFP positive cells in peripheral blood as well as in some erythroid tissues were decreased obviously. These results demonstrated that HS2 and HS3 elements would not show any synergy or superposition if they were located on different chromosomes in transgenic mice, and that HS2 element containing the core and flanking sequences might not be able to overcome the position effects and control position dependent expression of linked genes in transgenic mice. PMID- 18512118 TI - Terminal deoxynucleotidyl transferase negative T-cell lymphoblastic lymphoma in aleukemic patient. AB - Precursor lymphoblastic leukemia/lymphoblastic lymphoma (ALL/LBL) is a malignant neoplasm of precursor lymphocytes of T- or B-cell phenotype. We describe the unusual features of an ALL/LBL in an adolescent man in whom the disease presented with involvement of lymph nodes, but without bone marrow and peripheral blood involvement. Immunohistochemical studies revealed that the tumor cells were positive for CD3, CD34 class II, CD10, CD79a and CD99 but negative for TdT. Even though TdT was negative, he received ALL-therapy and is now in remission. PMID- 18512119 TI - QSID Tool: a new three-dimensional QSAR environmental tool. AB - QSID Tool (Quantitative structure-activity relationship tool for Innovative Discovery) was developed to provide an easy-to-use, robust and high quality environmental tool for 3D QSAR. Predictive models developed with QSID Tool can accelerate the discovery of lead compounds by enabling researchers to formulate and test hypotheses for optimizing efficacy and increasing drug safety and bioavailability early in the process of drug discovery. QSID Tool was evaluated by comparison with SYBYL using two different datasets derived from the inhibitors of Trypsin (Bohm et al., J Med Chem 42:458, 1999) and p38-MAPK (Liverton et al., J Med Chem 42:2180, 1999; Romeiro et al., J Comput Aided Mol Des 19:385, 2005; Romeiro et al., J Mol Model 12:855, 2006). The results suggest that QSID Tool is a useful model for the prediction of new analogue activities. PMID- 18512120 TI - Lymphangiogenesis induced by VEGF-C and VEGF-D promotes metastasis and a poor outcome in breast carcinoma: a retrospective study of 61 cases. AB - PURPOSE: To evaluate lymphangiogenesis in patients with breast carcinoma, explore the underlying mechanism, and study the relationship between lymphangiogenesis and progression of breast carcinoma. METHODS: Sixty-one cases of breast carcinoma with complete clinical and pathological data were analyzed. Using an anti podoplanin monoclonal antibody, an immunohistochemical study was made of all specimens to detect lymphatic vessel density (LVD) and to investigate its clinicopathological and prognostic value. VEGF-C and VEGF-D were observed by RT PCR and immunostaining to investigate their clinicopathological and prognostic values and their relationship with lymphangiogenesis. RESULTS: LVD in breast carcinoma (6.28+/-3.73) was significantly higher than in benign mammary lesions (0.50+/-1.27), P<0.01 and was significantly associated with lymphatic metastasis and high TNM stage, P<0.01. The level of VEGF-C and VEGF-D expression was also significantly higher in breast carcinomas than in benign mammary lesions, P<0.01. LVD increased significantly with higher expression of VEGF-C and VEGF-D, P<0.01. Patients with high expression of VEGF-C and VEGF-D were observed to be more likely to have a bad outcome, P<0.05. CONCLUSIONS: Lymphangiogenesis was significantly associated with lymph node metastasis, high TNM, and poor outcome in breast carcinoma. LVD may serve as a predictor of lymph node metastasis and a prognostic factor in breast carcinoma. VEGF-C and VEGF-D play an important role in lymphangiogenesis making the carcinoma more aggressive and leading to a poor prognosis. PMID- 18512121 TI - Dietary risk factors for squamous cell carcinoma of the upper aerodigestive tract in central and eastern Europe. AB - OBJECTIVE: The incidence of squamous cell carcinoma of upper aerodigestive tract (UADT: oral cavity, pharynx, larynx, and esophagus) has been increasing in central and eastern European countries. We investigated the relationship between diet and UADT cancers in these high risk areas. METHODS: We used data from hospital-based case-control study of 948 UADT cancer cases and 1,228 controls conducted in Romania, Hungary, Poland, Russia, Slovakia, and Czech Republic. Standardized questionnaire were used to collect information on 23 different food items, along with alcohol and tobacco consumptions. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for the UADT cancers after adjusting for center, age, sex, tobacco & alcohol intake, and other food groups. RESULTS: Consumption of dairy product was negatively associated with selected UADT cancers: larynx (OR: 0.38, CI: 0.23-0.62) and esophagus (OR: 0.55, CI: 0.33-0.93). While consumption of yellow/orange vegetables were inversely associated with oral/pharyngeal and laryngeal cancer (OR: 0.53, CI: 0.35-0.81 and OR: 0.62, CI: 0.38-1.00, respectively), preserved vegetable was positively associated with oral/pharyngeal and laryngeal cancer risk (p (trend) < 0.01 for both). CONCLUSION: Specific dietary components may play a role in the development of UADT cancers in the high-risk region of central and eastern Europe. PMID- 18512122 TI - General pattern of meiotic recombination in male dogs estimated by MLH1 and RAD51 immunolocalization. AB - The aim of this study was to estimate a general pattern of meiotic recombination in the domestic dog (Canis familiaris) using immunolocalization of MLH1, a mismatch repair protein of mature recombination nodules. We prepared synaptonemal complex (SC) spreads from 124 spermatocytes of three male dogs and mapped 4959 MLH1 foci along 4712 autosomes. The mean number of MLH1 foci for all autosomes was 40.0 foci per cell. Total recombination length of the male dog autosomal genome map was estimated as 2000 cM. A global pattern of MLH1 foci distribution along the autosomal bivalents was rather similar to that found in the mammals studied: a high frequency near the telomeres and a low frequency near the centromeres. An obligate MLH1 focus in the X-Y pairing region was usually located very close to Xp-Yq telomeres. The distances between MLH1 foci at autosomal bivalents were consistent with crossover interference. A comparison of the interference estimates coming from the distribution of MLH1 interfocus distances and RAD51/MLH1 focus ratio indicated a substantial variation between species in the strength of interference. PMID- 18512123 TI - Spatial and seasonal variation of water quality in an impacted coastal lagoon (Obidos Lagoon, Portugal). AB - The spatial distribution of silicate, ammonium, nitrate, nitrite, phosphate, chlorophyll a and dissolved oxygen in Obidos lagoon was obtained by surveying five sites in eight campaigns, between October 2004 and October 2006. A confined inner branch of the lagoon showed higher availability of ammonium (1.2-81 micromol l(-1)), phosphate (1.9-17 micromol l(-1)), silicate (0.85-86 micromol l( 1)) and chlorophyll a (0.30-18 microg l(-1)) than other sites (0.47-25 micromol l(-1), 0.10-3.9 micromol l(-1), 0.47-25 micromol l(-1), 0.25-11 microg l(-1), respectively). According to several trophic classification tools, that branch is considered eutrophic to polytrophic, emphasising its deteriorated conditions, while the rest of the lagoon is of better quality. In autumn/winter nutrients were inversely correlated to salinity (r > 0.93) reflecting the freshwater inputs enriched in nitrogen and phosphorous compounds to the inner branch. In warmer periods, dissolved oxygen concentrations dropped during the night, and sediments of the branch become an important source of ammonium and phosphate. The low DIN:P ratio (median = 10) obtained in the branch, which suggests an excess of phosphate, that increased in warmer periods and changed the limiting nutrient in the entire lagoon. These results emphasize the spatial heterogeneity of water quality in Obidos lagoon, its seasonal variability, and the importance of recognising these distributions before defining homogenous water body on the scope of Water Framework Directive. PMID- 18512124 TI - Source apportionment of PM10 and PM(2.5) at Tocopilla, Chile (22 degrees 05' S, 70 degrees 12' W). AB - Tocopilla is located on the coast of Northern Chile, within an arid region that extends from 30 degrees S to the border with Peru. The major industrial activities are related to the copper mining industry. A measurement campaign was conducted during March and April 2006 to determine ambient PM10 and PM(2.5) concentrations in the city. The results showed significantly higher PM10 concentrations in the southern part of the city (117 microg/m3) compared with 79 and 80 (microg/m3) in the central and northern sites. By contrast, ambient PM2.5 concentrations had a more uniform spatial distribution across the city, around 20 (microg/m3). In order to conduct a source apportionment, daily PM10 and PM(2.5) samples were analyzed for elements by XRF. EPA's Positive Matrix Factorization software was used to interpret the results of the chemical compositions. The major source contributing to PM(2.5) at sites 1, 2 and 3, respectively are: (a) sulfates, with approximately 50% of PM2.5 concentrations at the three sites; (b) fugitive emissions from fertilizer storage and handling, with 16%, 21% and 10%; (c) Coal and residual oil combustion, with 15%, 15% and 4%; (d) Sea salt, 5%, 6% and 16%; (e) Copper ore processing, 4%, 5% and 15%; and (f) a mixed dust source with 11%, 7% and 4%. Results for PM10--at sites 1, 2 and 3, respectively--show that the major contributors are: (a) sea salt source with 36%, 32% and 36% of the PM10 concentration; (b) copper processing emissions mixed with airborne soil dust with 6.6%, 11.5% and 41%; (c) sulfates with 31%, 31% and 12%; (d) a mixed dust source with 16%, 12% and 10%, and (e) the fertilizer stockpile emissions, with 11%, 14% and 2% of the PM10 concentration. The high natural background of PM10 implies that major reductions in anthropogenic emissions of PM10 and SO2 would be required to attain ambient air quality standards for PM10; those reductions would curb down ambient PM(2.5) concentrations as well. PMID- 18512125 TI - Evaluation of chromium contamination in water, sediment and vegetation caused by the tannery of Jijel (Algeria): a case study. AB - In order to evaluate the chromium (Cr) contamination due to the discharge of waste waters from the tannery of Jijel in the Mouttas river (Algeria), samples of water, sediment and vegetation (Agropyrum repens) were collected during a 6 month period in four stations located upstream (control) and downstream of the tannery. The total chromium was measured by atomic absorption spectrophotometry. Metal inputs were clearly related to effluent discharges from the tannery into the river. Although only traces of chromium were found in water samples upstream of the tannery, very high concentrations (up to 860 times higher) were detected downstream. The contamination was not limited to water of Mouttas River because a same difference in chromium concentrations was also found in sediments and plants Agropyrum repens that were sampled upstream and downstream of the tannery. This work showed that the treatment process used in the wastewater treatment plant of the tannery of Jijel is not able to remove the chromium detected in their influents. The occurrence and chromium levels detected in the aquatic environment represent a major problem concerning drinking water resources and environmental protection of water bodies. PMID- 18512126 TI - Application of neural-based modeling in an assessment of pollution with mercury in the middle part of the Warta River. AB - The level of pollution with various mercury species (organomercury, water- and acid-soluble mercury, mercury bound to humic matter and to sulphides) of the floodplain soils and sediments from middle part of the Warta River has been assessed using self-organizing maps (SOM). Chemometric evaluation allowed identification of moderately (median 173-187 ng g(-1), range 54-375 ng g(-1) in soil and 130 ng g(-1), range 47-310 ng g(-1) in sediment) and heavily polluted samples (662 ng g(-1), range 426-884 ng g(-1)). Heavily polluted were located mainly below and in the area of the Poznan city. Statistical comparison of mercury species distribution in floodplain soils of the Warta River shows different patterns for moderately and heavily polluted samples. In heavily polluted soils the contribution of mobile mercury (sum of organomercury species, water- and acid soluble species) is lower (4.2%) than in moderately polluted soils (6.1%). Higher contribution of mobile mercury was observed in sediments of the Warta River (12%). In case of moderately polluted samples, statistical differences in the contribution of mercury species are relatively low and thus the environmental risk from mercury deposited in aquatic system of the Warta River is relatively low. However, higher water levels and heavy floods may incite remobilisation of some organomercuries (2.2-2.9 ng g(-1) in soil and 10 ng g(-1) in sediment) and acid-soluble species of mercury (2.6-2.9 ng g(-1) in soil and 0.5 ng g(-1) in sediment). PMID- 18512127 TI - Diammonium hydrogen phosphate as a versatile and efficient catalyst for the one pot synthesis of pyrano[2,3-d]pyrimidinone derivatives in aqueous media. AB - Diammonium hydrogen phosphate, (NH4)2HPO4, was used as a catalyst for one-pot, three-component condensation reactions consisting of aromatic aldehydes, malononitrile and barbituric/thiobarbituric acid in aqueous ethanol at room temperature. This method has the advantages of a simple operation, mild reaction conditions, high yields, by using a less toxic and low cost chemical as a catalyst. PMID- 18512128 TI - Right atrial function: still underestimated in clinical cardiology. PMID- 18512129 TI - Cryptococcus neoformans, Cryptococcus gattii: serotypes in Venezuela. AB - Cryptococcus neoformans is one of the medically important yeast-like fungi. C. neoformans var. gatti has been made a species: C. gatti. In our country, there are few studies about these two species and their serotypes. The aim of this study was to determine the distribution of C. neoformans and C. gattii, and their serotypes in Venezuelan clinical isolates. One hundred and twenty C. neoformans and 12 C. gattii clinical isolates were identified by L-canavanine, glycine, and bromothymol blue agar media (CGB). These were investigated by agglutination and adsorption studies with anticryptococcal sera, which were produced by rabbit immunization. Of the 132 isolates 59.8% were typed serotype A (C. neoformans), followed by 25.8% serotype D (C. neoformans), 5.3% serotype AD (C. neoformans), and 5.3% were typed serotype C (var. gattii). Additionally 3.8% were serotype B (C. gattii). PMID- 18512130 TI - Breakthrough disseminated Scedosporium prolificans infection in a patient with relapsed leukaemia on prolonged voriconazole followed by posaconazole prophylaxis. AB - A man with acute lymphoblastic leukaemia developed disseminated Scedosporium prolificans infection following chemotherapy for disease relapse while on posaconazole prophylaxis. Scedosporium prolificans infection during posaconazole prophylaxis has not been reported previously. This report is timely as the uptake of posaconazole, the broadest spectrum azole clinically available, is likely to grow with recent evidence supporting its role as prophylaxis against invasive fungal infections in high-risk haematology patients. PMID- 18512131 TI - Association of polymorphisms at restriction enzyme recognition sites of apolipoprotein B and E gene with dyslipidemia in children undergoing primary nephrotic syndrome. AB - BACKGROUND: Dyslipidemia, a common complication, is very prevalent in children with primary nephrotic syndrome (PNS). Recent studies have shown that genetic basis may be involved in the onset of HLP secondary to PNS. ApoB and E have been identified as the important candidate genes for lipid abnormalities. OBJECTIVE: To investigate the association of apolipoprotein B (apoB) and E (apoE) genetic polymorphisms (Xba I, EcoR I, Msp I, and Hha I) with parameters describing the serum lipid profiles in children undergoing PNS. METHODS: Genomic DNA was extracted from 250 children diagnosed with PNS and 200 healthy controls with neither allergic nor renal disease. ApoB (Xba I, EcoR I, and Msp I) and apoE (Hha I) genotypes were determined by PCR-restriction fragment length polymorphism (RFLP) analysis. The fasting serum lipoprotein (a) [Lp(a)], total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (apoA1), apoB, and total protein from a 24-h urine sample were measured. RESULTS: No significant differences in genotypes and alleles frequencies were observed for the apoB Xba I, EcoR I, Msp I and the apoE Hha I restriction sites in PNS patients as compared to controls (P > 0.05). Patients and controls with X + allele exhibited significantly higher serum levels of Lp(a), TC, nonHDL-C, LDL-C, LDL-C/HDL-C ratio, and apoB than that with X- allele (P < 0.05), whereas for apoA1/B ratio the opposite was found (P < 0.01). E-/E- carriers had significantly higher Lp(a), TC, HDL-C, and apoA1 concentrations than did E+/E- or E+/E+ carriers in control group (P < 0.05). Healthy children carrying the rare EcoR I allele had higher mean Lp(a), TC, and HDL-C levels than homozygotes for E+ (P < 0.05). Higher Lp(a) serum concentrations were observed in patients with E- allele (P < 0.05). No significant differences in lipid parameters were determined for the apoB Msp I and apoE Hha I the polymorphisms study (P > 0.05). When genetic variations were compared with urinary protein excretion, the Xba I X- allele was more frequent in patients with elevated proteinuria (P < 0.01). CONCLUSION: Presence of Xba I X+ allele and/or EcoR I E- at the apoB gene may be risk factors for lipid abnormalities secondary to childhood PNS. PMID- 18512132 TI - Negotiated or taken-for-granted trust? Explicit and implicit interpretations of trust in a medical setting. AB - Trust between a patient and a medical doctor is normally both justified and taken for granted, but sometimes it may need to be negotiated. In this paper I will present how trust can be interpreted as both an explicit and implicit phenomenon, drawing on literature from the social sciences and philosophy. The distinction between explicit and implicit interpretations of trust will be used to address problems that may arise in clinical consultations. Negotiating trust in any way very easily brings distrust into a situation, but sometimes this can be helpful for building a more functional patient-doctor relationship. PMID- 18512133 TI - Near miss, gambler's fallacy and entrapment: their influence on lottery gamblers in Thailand. AB - Near miss, gambler's fallacy and entrapment are hypothesized to influence gambling behavior but their individual impact on gambling behavior and their relationships have not been investigated. This study examines the direct and mediated effects of near miss, gambler's fallacy and entrapment on lottery gambling among Thai lottery gamblers. Two groups of 200 lottery gamblers participated in the study. One group used superstitious methods to search for lottery numbers to bet, whereas the other group simply bought their tickets from lottery stalls. The superstitious group shows higher levels of near miss, gambler's fallacy, entrapment and lottery gambling. Three models of relationships among near miss, gambler's fallacy, entrapment and lottery gambling were constructed using LISREL program. The model that shows a direct effect of near miss, and one that shows a mediated effect of near miss fit the data equally. The results confirm a gambler's fallacy and entrapment as starting points in the theory of cognitive psychology of lottery gambling and the generalization of near miss in gambling motivation. PMID- 18512134 TI - Pharmacological probing of type 1 autism. PMID- 18512135 TI - A meta-analysis of sensory modulation symptoms in individuals with autism spectrum disorders. AB - Sensory modulation symptoms are common in persons with autism spectrum disorders (ASD); however have a heterogeneous presentation. Results from 14 studies indicated a significant high difference between ASD and typical groups in the presence/frequency of sensory symptoms, with the greatest difference in under responsivity, followed by over-responsivity and sensation seeking. Three moderators that reduced the variability in findings among studies were: chronological age, severity of autism, and type of control group. Sensory differences were highest for studies of children ages 6-9 years, samples with more than 80% with an autism diagnosis, and compared to a CA matched versus a MA or DD matched group. It is important to consider these moderators in the design of studies and interventions addressing sensory symptoms. PMID- 18512136 TI - Abnormal transmethylation/transsulfuration metabolism and DNA hypomethylation among parents of children with autism. AB - An integrated metabolic profile reflects the combined influence of genetic, epigenetic, and environmental factors that affect the candidate pathway of interest. Recent evidence suggests that some autistic children may have reduced detoxification capacity and may be under chronic oxidative stress. Based on reports of abnormal methionine and glutathione metabolism in autistic children, it was of interest to examine the same metabolic profile in the parents. The results indicated that parents share similar metabolic deficits in methylation capacity and glutathione-dependent antioxidant/detoxification capacity observed in many autistic children. Studies are underway to determine whether the abnormal profile in parents reflects linked genetic polymorphisms in these pathways or whether it simply reflects the chronic stress of coping with an autistic child. PMID- 18512137 TI - Brief report: narratives of personal events in children with autism and developmental language disorders: unshared memories. AB - Narrative analysis of personal events provides an opportunity for identifying autism specific issues related to language and social impairments. Eight personal events were elicited from three groups of school age children: 14 high functioning with Autism Spectrum Disorders (HFA), 12 non-autistic with developmental language disorders (DLD), and 12 typically developing matched for chronological age and non-verbal IQ. The coding focused on narrative format (constituents) and style (coherence). The analyses indicate basic knowledge of conventional narrative format in all groups but a consistent lack of high-point in HFA children's stories interpreted as a consequence of their lack of social understanding of narrative. The results suggest novel interventions to foster autobiographical memory in HFA children which may assist in their self-awareness development. PMID- 18512138 TI - A non-radioactive PCR-SSCP analysis allows to distinguish between HPV 16 European and Asian-American variants in squamous cell carcinomas of the uterine cervix in Colombia. AB - Human Papillomavirus type 16 (HPV 16) DNA is regularly found in around 50% of all cervical carcinomas. Variants of this type have been found associated with different risks for cervical cancer development. Presence of HPV 16 variants in Colombia has not been previously reported. The aims of this study were to assess the feasibility of non-radioactive PCR-SSCP (polymerase chain reaction single strand conformation polymorphism) analysis for determination of variability of ORF of E6, variability in the enhancer sequence of the LCR, and for establishment of the distribution of HPV 16 variants in invasive squamous cell carcinoma of the uterine cervix in Colombian women. Biopsies from 59 patients at the Instituto Nacional de Cancerologia (INC) in Bogota (Colombia) were collected. HPV detection was performed using universal primers. HPV 16 variants were detected by non radioactive single-stranded conformational polymorphism (SSCP) analysis and direct sequencing. HPV 16 was detected in 57.6% of the tumors. The European branch was identified in 88.2% of the samples with the E-G350 class being the most prevalent variant (41.1%). The Asian-American branch was identified in 8.8% of the samples. Within this group it was possible to distinguish between c and a classes. It was not possible to determine the branch in 2.9% of the cases. A nucleotide transition (G to A) at position 7521 was the most prevalent variation (80%) found in the enhancer sequence of the LCR region. CONCLUSION: A non radioactive PCR-SSCP analysis allowed us to distinguish between European and Asian-American branches of HPV 16, and to distinguish among classes in squamous cell carcinomas of the uterine cervix in Colombia. This method is an excellent alternative that can be used as a screening tool for identification of HPV 16 variants. PMID- 18512139 TI - Partial sequence analysis of VP1 of Indian isolates of foot-and-mouth disease virus type Asia-1. AB - Nucleotide sequence of 3' end of VP1 (1D region) was determined using RT-PCR amplified DNA of 31 foot and mouth disease virus (FMDV) type Asia-1 field isolates originating from 11 different geographically distinct states of India during the period 1987-2000. These field strains exhibited an average of 7.5% divergence among them and were found to be divergent from the Indian vaccine strains Asia-1 WBN 117/85, IND 8/79, and IND 63/72, by an average 5.9, 14.8, and 7.4% divergence, respectively. Phylogenetic analysis of these 31 field isolates including 3 of the vaccine strains of India and sequences of 22 Indian field isolates obtained from the GenBank revealed that all the Indian FMDV type Asia-1 isolates belonged to a single genotype comprising of two distinct lineages (Lineages A and B). All the field isolates under study belonged to the Lineage-B comprising 8 different clusters, which also includes the vaccine strains WBN 117/85 and IND 8/79. Surprisingly, another vaccine strain IND 63/72 formed Lineage-A. Phylogenetic analysis of sequences of another 23 exotic type Asia-1 isolates from 15 different countries obtained from the GenBank along with the 56 Indian isolates revealed the existence of three distinct genotypes. The prototype strain Asia-1 PAK 1/54 belongs to a separate genotype. Two strains from India along with one strain each from China and Russia belongs to another genotype. The third genotype is formed by the remaining isolates including all the 31 isolates from the present study and exotic viruses from 14 other different countries. Comparison of deduced amino acid (aa) sequence indicated that majority of the mutations were found within two distinct regions corresponding to amino acid positions 130-160 and 193-211. The motif at aa positions 138-141 in vaccine strains WBN 117/85, IND 8/79 and in all the field isolates was ETTS/P; however, the same motif in IND 63/72 was TQPT. The motif 153-156 in majority of Indian isolates including vaccine strains WBN 117/85 and IND 8/79 was LSGQ/R whereas the same motif seen in IND 63/72 was VSNR. The study revealed that the FMDV type Asia 1 isolates circulating in the country are not highly heterogeneous, but showed considerable genetic variations. Certain mutations were also observed in the residues, which have been proved to be contributing to the formation of neutralizing epitopes. In neutralization studies employing polyclonal antisera, type Asia-1 WBN 117/85 revealed broader serological spectrum than other vaccine strains of India used in this study. PMID- 18512140 TI - Beyond the Down Low: sexual risk, protection, and disclosure among at-risk Black men who have sex with both men and women (MSMW). AB - Little information is available about sexual risk, protective, and disclosure practices among Black bisexually active men and how these may be amenable to intervention when necessary. In-depth interviews were conducted with 30 at-risk urban Black men who have sex with both men and women (MSMW). Participants reported protecting themselves and their partners through routine HIV testing, using condoms consistently, engaging in strategic positioning during sexual activity, and limiting the number of sexual partners. In addition, they described several reasons for not using protection, including not having condoms available, enjoying sexual activity more without condoms, and perceiving female partners to be "safer" than male partners. Disclosure of bisexuality was complex and reportedly difficult, in particular to female and gay-identified male partners. Future interventions must not only build protection and disclosure skills among MSMW, but also increase broader social awareness and acceptance of male bisexuality. PMID- 18512141 TI - 100% adherence study: educational workshops vs. video sessions to improve adherence among ART-naive patients in Salvador, Brazil. AB - We conducted a randomized trial to test an intervention aimed at increasing adherence to antiretroviral therapy (ART) among HIV-positive, ART-naive patients in Salvador, Brazil. Participants (N = 107) were randomized to either educational workshops based on the information-motivation-behavioral skills model (n = 52) or a control video session (n = 55). Changes in self-reported ART adherence, viral load, CD4 cell counts and ART pharmacy records were measured periodically over 12 months. After 3-6 months, ART adherence (> or = 95%) was 77.8% in the workshop group and 85.7% in video group (as treated) and 53.8% and 65.5%, respectively, using intention-to-treat (ITT) analysis (both P > 0.05) At 9-12 months, ART adherence decreased to 73.7% in the workshop group and 79.1% in the video group (as treated) and 53.8% and 61.8% using ITT, respectively. No differences were found in self-reported adherence, viral load or pharmacy records between groups. We found that the educational workshop intervention does not increase adherence to ART. PMID- 18512142 TI - Vengeance, HIV disclosure, and perceived HIV transmission to others. AB - Feelings of vengefulness result from being treated unfairly. However, some individuals are more sensitive to unfair treatment and more likely to demand restitution than others. Degrees of vengefulness may influence behavior in HIV positive men who have sex with men (MSM), where highly vengeful men may seek limited retribution by placing others at risk, for example, by failing to disclose their HIV-status to sexual partners. This study examined the tendency towards vengefulness in HIV-positive MSM and its associations with disclosure and condom use behaviors. Results showed that greater certainty of from whom participants had contracted HIV was associated with lowered vengefulness over time. Though condom use did not vary by vengefulness, MSM reporting higher vengefulness concealed their HIV serostatus more than men reporting less vengefulness. Vengeance was not related to individuals' perceptions that they had transmitted the disease to others. Overall, the data suggested identifying one's HIV transmitter was reconciliatory. Men reporting higher vengefulness might also derive a sense of justice from not disclosing their serostatus. PMID- 18512143 TI - HIV risk and the internet: results of the Men's INTernet Sex (MINTS) Study. AB - This study assessed the feasibility of online recruitment of high-risk Latino men who have sex with men (MSM) for HIV prevention survey research and investigated the relationship between Internet use and unsafe sex. Participants (N = 1,026) were Internet-using Latino MSM living in the U.S. recruited using online banner advertisements. Respondents completed a cross-sectional, online survey in English or Spanish. Sample characteristics reflected national statistics within 5%. Nearly all (99%) reported having used the Internet to seek sex with another man. Two-thirds of respondents reported having unprotected anal sex with > or =1 man in the last year, 57% of these with multiple partners. Participants reported engaging in anal sex and unprotected anal sex with nearly twice as many men first met online versus offline, but risk proportions did not differ. Internet-based HIV prevention research is possible even with geographically-dispersed minority populations. Efficiency appears the primary risk associated with meeting partners online. PMID- 18512144 TI - Travel distance and the use of inpatient care among patients with schizophrenia. AB - This study examines the variations in the use of inpatient care that can be explained by travel distance among patients with schizophrenia living in Taiwan. Data were drawn from the Psychiatric Inpatient Medical Claims Database. We used mediation analysis and multilevel analysis to identify associations. Travel distance did not significantly account for lower readmission rates after an index admission, but significantly explained the longer length of stay of an index admission by 9.3 days (P<0.001, 85% of variation) between remote and non-remote regions. Policies are discussed aimed at reducing the impact of travel distance on rural mental health care through inter-disciplinary collaboration and telepsychiatry. PMID- 18512145 TI - When programs benefit some people more than others: tests of differential service effectiveness. AB - Practitioners need to know for whom evidence-based services are most or least effective, but few services research studies provide this information. Using data from a randomized controlled comparison of supported employment findings for two multi-service psychiatric rehabilitation programs, we illustrate and compare procedures for measuring program-by-client characteristic interactions depicting differential program effectiveness, and then illustrate how a significant program by-client interaction can explain overall program differences in service effectiveness. Interaction analyses based on cluster analysis-identified sample subgroups appear to provide statistically powerful and meaningful hypothesis tests that can aid in the interpretation of main effect findings and help to refine program theory. PMID- 18512146 TI - Genetic polymorphism of type 1 intermediate filament wool keratin gene in native Indian sheep breeds. AB - Information is presented on the genetic polymorphism of the Type 1 intermediate filament wool keratin gene in 15 native Indian sheep breeds belonging to different agro-ecological regions of India. The study analyzed random blood samples of the 638 sheep by the PCR-RFLP technique. Restriction digestion analysis of a 480 bp PCR fragment of the first exon region with MspI revealed two allelic variants (M = 0.748 and N = 0.252) and three genotypes (MM = 0.543, MN = 0.410, and NN = 0.047) across the 15 sheep breeds. The allelic frequency differences for both alleles across the Indian breeds, irrespective of their geographic distribution, color pattern, and utility traits, were observed to be statistically insignificant by a chi-square test (P > 0.05). According to the pattern of occurrence of allelic variants (M > N), the Indian breeds exhibited similarity to some of the reported European sheep breeds. The average heterozygosity was 0.420, and none of the breeds deviated from Hardy-Weinberg equilibrium. The predominance of the M over the N allele supported its ancestry in Indian sheep too. PMID- 18512147 TI - Enhanced survival of melanopsin-expressing retinal ganglion cells after injury is associated with the PI3 K/Akt pathway. AB - In the present study, we studied the factors that contribute to the injury resistant property of melanopsin-expressing retinal ganglion cells (mRGCs). Since phosphatidylinositol-3 kinase (PI3 K)/Akt signaling pathway is one of the well known pathways for neuronal cell survival, we investigated the survival of mRGCs by applying the PI3 K/Akt specific inhibitors after injury. Two injury models, unilateral optic nerve transection and ocular hypertension, were adopted using Sprague-Dawley rats. Inhibitors of PI3 K/Akt were injected intravitreally following injuries to inhibit the PI3 K/Akt signaling pathway. Retinas were dissected after designated survival time, immunohistochemistry was carried out to visualize the mRGCs using melanopsin antibody and the number of mRGCs was counted. Co-expression of melanopsin and phospho-Akt (pAkt) was also examined. Compared to the survival of non-melanopsin-expressing RGCs, mRGCs showed a marked resistance to injury and co-expressed pAkt. Application of PI3 K/Akt inhibitors decreased the survival of mRGCs after injury. Our previous study has shown that mRGC are less susceptible to injury following the induction of ocular hypertension. In this study, we report that mRGCs were injury-resistant to a more severe type of injury, the optic nerve transection. More importantly, the PI3 K/Akt pathway was found to play a role in maintaining the survival of mRGCs after injury. PMID- 18512148 TI - BRCA1 germline mutations and tumor characteristics in Chinese women with familial or early-onset breast cancer. AB - The data related to BRCA1 germline mutation in Chinese women with familial breast cancer is increasing. However, little is known the frequency of BRCA1 mutations in Chinese women with familial or early-onset breast cancer from Northern China, and few studies are available to investigate the clinicopathological characteristics of BRCA1 tumors in Chinese women. In this study, we detected germline mutations in BRCA1 in a cohort of 139 breast cancer patients who either have a family history of breast cancer (n = 68) or whose tumors are diagnosed at or before the age of 35 (n = 71) from Northern China. A total of 6 deleterious BRCA1 mutations were identified in this cohort, 4 of which (5587-1 del8, 3887 del AG, IVS21 + 1delG, and 2129 ins TG) are novel and one mutation (3478del5) detected in this study was only reported in Chinese population. The frequency of BRCA1 mutations in women with familial or early-onset breast cancer was 5.9% (4/68) or 2.8% (2/71) in this cohort, respectively; but the mutations were detected in 4 of 16(25.0%) familial breast cancer patients whose tumors were diagnosed before the age of 40. Moreover, BRCA1 mutation tumors tended to be high histological grade, and to be negative for ER, PgR, and Her-2 compared with tumors without BRCA1 mutations. Our study suggests that Chinese women with a family history of breast cancer whose tumors are diagnosed before age of 40 would be a suitable candidate for BRCA1 testing; and BRCA1 tumors in Chinese women exhibit an aggressive phenotype. PMID- 18512149 TI - The expression patterns of beta1,4 galactosyltransferase I and V mRNAs, and Galbeta1-4GlcNAc group in rat gastrocnemius muscles post sciatic nerve injury. AB - Glycosylation is one of the most important post-translational modifications. It is clear that the single step of beta1,4-galactosylation is performed by a family of beta1,4-galactosyltransferases (beta1,4-GalTs), and that each member of this family may play a distinct role in different tissues and cells. beta1,4-GalT I and V are involved in the biosynthesis of N-linked oligosaccharides and play roles in sciatic nerve regeneration after sciatic nerve injury. In the present study, the expression of beta1,4-galactosyltransferase (beta1,4-GalT) I, V mRNAs and Galbeta1-4GlcNAc group were examined in rat gastrocnemius muscles after sciatic nerve crush and transection. Real time PCR revealed that beta1,4-GalT I and V mRNAs expressed at a high level in normal gastrocnemius muscles and decreased gradually from 6 h, reached the lowest level at 2 weeks, then restored gradually to relatively normal level at 4 weeks after sciatic nerve crush. In contrast, in sciatic nerve transection model, beta1,4-GalT I and V mRNAs decreased gradually from 6 h, and remained on a low level at 4 weeks in gastrocnemius muscles after sciatic nerve transection. In situ hybridization indicated that beta1,4-GalT I and V mRNAs localized in numerous myocytes and muscle satellite cells under normal conditions and at 4 weeks after sciatic nerve crush, and in a few muscle satellite cells at 4 weeks after sciatic nerve transection. Furthermore, lectin blotting showed that the expression level of the Galbeta1-4GlcNAc group decreased from 6 h, reached the lowest level at 2 weeks, and restored to relatively normal level at 4 weeks after sciatic nerve crush. RCA I lectin histochemistry demonstrated that Galbeta1-4GlcNAc group localized in numerous membranes of myocytes and muscle satellite cells in normal and at 4 weeks after sciatic nerve crush, and in a few muscle satellite cells at 2 and 4 weeks after sciatic nerve transection. These results indicated that the expressions of beta1,4-GalT I, V mRNAs and Galbeta1-4GlcNAc group were involved in the process of denervation and reinnervation, which suggests that beta1,4-GalT I, V mRNAs and Galbeta1-4GlcNAc group may play an important role in the muscle regeneration. PMID- 18512150 TI - Behavioral and electrophysiological evidence for the differential functions of TRPV1 at early and late stages of chronic inflammatory nociception in rats. AB - We previously reported that vanilloid receptor type 1 (VR1, or TRPV1) was up regulated in dorsal root ganglion (DRG) and the spinal dorsal horn after chronic inflammatory pain produced by complete Freund's adjuvant (CFA) injection into the plantar of rat hind paw. In the present study, we found that subcutaneous or intrathecal application of capsazepine (CPZ), a TRPV1 competitive antagonist, could inhibit thermal hyperalgesia on day 1 and on day 14 but not on day 28 after CFA injection. With extracellular electrophysiological recording, the effect of CPZ on noxious electrical or heat stimulation evoked responses of wide dynamic range (WDR) neurons in the deep layers of the spinal dorsal horn was evaluated. Under noxious electrical stimulation to sciatic nerve, CPZ applied to the spinal cord produced an inhibition on Adelta- and C-fiber evoked responses of WDR neurons on day 1 and 14, but not on day 28. Under radiant heat stimulation to the receptive field skin, subcutaneous application of CPZ significantly inhibited the background activity and extended the response latency of WDR neurons on day 14. These results provide new evidence for the functional significance of TRPV1 at the early stage, but not the late stage, in the rat model of CFA-induced inflammatory pain. PMID- 18512151 TI - Cytoprotective effect of Valeriana officinalis extract on an in vitro experimental model of Parkinson disease. AB - Parkinson's disease (PD) is one of the most important neurodegenerative worldwide disorders. The potential cytoprotective effects of aqueous extract of Valeriana officinalis on rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells were demonstrated. The cytotoxicity, cell viability and analysis of cellular morphology were performed by MTT-tetrazole (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay and phase contrast microscopy, respectively. Significant changes in the cellular morphology, and condensation of the cell body could be observed when cells were treated with 300 nM rotenone for 48 h. Three different concentrations of Valeriana officinalis extract were used (0.049, 0.098 and 0.195 mg/mL). These extracts brought about an increase of 7.0 +/- 1.3%, 14.5 +/- 1.3% and 14.5 +/- 3.2% in cell viability. Our results indicated that neuroprotector action of the Valeriana officinalis extract provides support for later studies as they help understanding this drug for the development of cytoprotective various therapies in PD. PMID- 18512152 TI - Initiation of oligodendrocyte progenitor cell migration by a PDGF-A activated extracellular regulated kinase (ERK) signaling pathway. AB - During CNS development, oligodendrocyte progenitor (OP) cells migrate from germinal zones to presumptive white matter tracts to generate myelinating oligodendrocytes. In vitro and in vivo studies indicate that platelet-derived growth factor-A (PDGF-A) is a potent chemoattractant for OP cells and important for normal distribution throughout the developing CNS. However, PDGF-A does not localize in concentration gradients corresponding to OP migratory pathways, as would be expected for a chemoattractant to direct migration. Therefore, the mechanism by which PDGF-A regulates OP distribution remains to be clarified. Here we show that PDGF-A induces OP migration and continuous exposure to PDGF-A is not required to maintain migration. Using pharmacological inhibitors, we show that a self-sustaining extracellular-regulated-kinase signaling pathway drives OP migration for up to 72 hours after the initial PDGF stimulus. These findings indicate PDGF-A may act to mobilize OP cells that then respond to distinct directional signals to distribute appropriately within the CNS. PMID- 18512154 TI - Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP. AB - Two-photon fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue. We evaluated and optimized the FRET pair of mEGFP (monomeric EGFP with the A206K mutation) and REACh (non-radiative YFP variants) for TPFLIM. We characterized several mutants of REACh in terms of their "darkness," and their ability to act as a FRET acceptor for mEGFP in HeLa cells and hippocampal neurons. Since the commonly used monomeric mutation A206K increases the brightness of REACh, we introduced a different monomeric mutation (F223R) which does not affect the brightness. Also, we found that the folding efficiency of original REACh, as measured by the fluorescence lifetime of a mEGFP-REACh tandem dimer, was low and variable from cell to cell. Introducing two folding mutations (F46L, Q69M) into REACh increased the folding efficiency by approximately 50%, and reduced the variability of FRET signal. Pairing mEGFP with the new REACh (super-REACh, or sREACh) improved the signal-to-noise ratio compared to the mEGFP-mRFP or mEGFP-original REACh pair by approximately 50%. Using this new pair, we demonstrated that the fraction of actin monomers in filamentous and globular forms in single dendritic spines can be quantitatively measured with high sensitivity. Thus, the mEGFP-sREACh pair is suited for quantitative FRET measurement by TPFLIM, and enables us to measure protein-protein interactions in individual dendritic spines in brain slices with high sensitivity. PMID- 18512153 TI - Sorafenib triggers antiproliferative and pro-apoptotic signals in human esophageal adenocarcinoma cells. AB - BACKGROUND AND PURPOSE: Current therapies offer scant benefit to patients with advanced esophageal adenocarcinoma. We investigated the effects of Sorafenib, a multifunctional kinase inhibitor, on several growth regulatory pathways that control cell growth and survival in SEG-1 cells derived from Barrett's adenocarcinoma. METHODS: SEG-1 cells were exposed to acidified medium or taurocholic acid, with and without pre-incubation with Sorafenib. Cyclin D1 and E, c-Myc, and Bcl-2 expression levels as well as STAT3 activations were determined by Western blotting. Cyclin D1 mRNA was measured by real-time PCR. Apoptosis was assessed by TUNEL assay. RESULTS: Sorafenib significantly inhibited SEG-1 cell proliferation stimulated by acid or bile acid treatments and reduced cell survival. This drug significantly reduced the up-regulations of cyclin D1, cyclin E, c-Myc, and Bcl-2 as well as the activation of STAT3 in SEG-1 cells. CONCLUSIONS: These results support a rational basis for future clinical studies to assess the therapeutic benefit of Sorafenib in esophageal adenocarcinoma. PMID- 18512155 TI - Expenditures in mental illness and substance use disorders among veteran clinic users with diabetes. AB - Few studies have looked at the health-care expenditures of diabetes patients based on the type of co-occurring conditions of mental illness (MI) or substance use disorders (SUD). Our study analyzes the health-care expenditures associated with various diagnostic clusters of co-occurring drug, alcohol, tobacco use, and mental illness in veterans with diabetes. We merged Veteran Health Administration and Medicare fee-for-service claims database (fiscal years 1999 and 2000) for analysis (N = 390,253) using generalized linear models; SUD/MI were identified using International Classification of Diseases, 9th edition codes. The total average expenditures (fiscal year 2000) were lowest ($6,185) in the "No MI and No SUD" and highest ($19,801) for individuals with schizophrenia/other psychoses and alcohol/drug use. High expenditures were associated with both SUD and MI conditions in diabetes patients, and veterans with alcohol/drug use had the highest expenditures across all groups of MI. These findings reinforce the need to target groups with multiple comorbidities specifically those with serious mental illnesses and alcohol/drug use for interventions to reduce health-care expenditures. PMID- 18512156 TI - Mental health and substance abuse spending by age, 2003. AB - This article presents national estimates of mental health and substance abuse (MHSA) spending in 2003 by age groups. Overall, $121 billion was spent on MHSA treatment across all age groups in 2003. Of the total $100 billion spent on MH treatment, about 17% was spent on children and adolescents, 68% on young and mid age adults, and 15% on older adults. MH spending per capita by age was $232 per youth, $376 per young and mid-age adult, and $419 per older adult. Of the total $21 billion spent on SA treatment, about 9% was spent on children and adolescents, 86% on adults ages 18 through 64, and 5% on older adults age 65 and older. SA spending per capita by age was $26 per youth, $98 per mid-age adult, and $28 per older adult. PMID- 18512157 TI - Money, case complexity, and wait lists: perspectives on problems and solutions at children's mental health centers in Ontario. AB - Senior managers of children's mental health centers across Ontario, Canada were interviewed regarding the challenges and solutions of access and delivery of care. The central challenges--funding, case complexity, waitlists, staffing, and system integration--revealed a complex interplay between the policies and financing of children's mental health services and the provision of clinical services at the agency level and within the community. The desire for integration and collaboration was countered by competition for funding and service demands. A need for policies that allow for local solutions while providing leadership for sustained improvements in the ease and timeliness of access to care and effective clinical services emerged. PMID- 18512158 TI - Is it wraparound yet? Setting quality standards for implementation of the wraparound process. AB - The wraparound process has increasingly been used as a mechanism to plan and coordinate services for children with behavioral health needs and their families. This has led to growing interest in assessing wraparound implementation against standards for quality. However, there has been little consideration of how best to establish such benchmarks or guidelines. Using both a norm-referenced and criterion-referenced approach, this study established preliminary criteria for assessing the adequacy of wraparound implementation using the Wraparound Fidelity Index, version 3, a multi-informant interview that assesses conformance to wraparound principles. The evaluation system was then applied to ten wraparound programs and 11 different study samples assessed via the Wraparound Fidelity Index version 3 in research studies. The system was constructed to discriminate different wraparound conditions assessed in research studies while still being attainable by the ten established wraparound programs. Implications for evaluating wraparound programs and for setting fidelity benchmarks in behavioral health services research are discussed. PMID- 18512159 TI - Scientific perspectives on extending the provision for waivers of in vivo bioavailability and bioequivalence studies for drug products containing high solubility-low permeability drugs (BCS-Class 3). AB - Recently, there has been increased interest in extending the provision for waivers of in vivo bioavailability and bioequivalence (BA-BE) studies that appeared in the guidance published by the Food and Drug Administration (FDA) (1) to pharmaceutical products containing Class 3 drugs (High solubility-Low Permeability). The extension of the Biopharmaceutics Classification System (BCS) to Class 3 drugs is meritorious because of its impact on public health policy considerations. The rate limiting step in the absorption of Class 3 drugs is the permeability through the intestinal membrane. This commentary will focus its attention on the scientific considerations which need to be examined to assess the risk and the benefit prior to granting a waiver of in vivo bioavailability and/or bioequivalence studies for Class 3 drugs. It will examine the forces affecting the interconnectivity of the neuronal, immunological and hormonal systems in the gastrointestinal tract that may affect its permeability and functionality. It will also challenge the assumption that in vitro dissolution and in vitro permeability studies in tissue cultures in the presence and absence of excipients are good predictors for in vivo dissolution and in vivo permeability which are at the heart of the BCS. PMID- 18512160 TI - Microglial cells and Parkinson's disease. AB - Chronic inflammation mediated by microglial cells is the fundamental process contributing to the death of dopamine (DA)-producing neurons in the brain. Production of inflammatory products by these microglial cells characterizes the slow destructive process in Parkinson's disease (PD). The activation of microglial cells and the generation of pro-inflammatory cytokines that characterize PD are mediated by several different signaling pathways, with the activation of the respiratory burst by microglial cells being a critical event in the ultimate toxicity of DA-neurons. The work on our lab is concerned with understanding the mechanisms of activation, response, and therapeutic targets of microglial cells, with the aim to provide more effective treatments for PD and other inflammatory diseases of the CNS. PMID- 18512161 TI - Isolation and biomass production of a Saccharomyces cerevisiae strain binding copper and zinc ions. AB - Copper and zinc are essential trace elements participating in many physiological functions, notably immunity and protection against oxidative stress. Yeasts and the yeast Saccharomyces cerevisiae, in particular, possess in their genome tandem repeats of the CUP1 gene coding for a protein (a metallothionein) capable of capturing and binding toxic elements such as copper ions. The number of copies of this gene in a cell determines its physiological level of resistance to these ions. This paper describes the selection, characterization, and production of a new copper-resistant yeast strain that can bind large quantities of copper and zinc. This approach should lead to increasing the bioavailability of these trace elements and hence to reducing their emission into the environment. PMID- 18512162 TI - Model-based fed-batch for high-solids enzymatic cellulose hydrolysis. AB - While many kinetic models have been developed for the enzymatic hydrolysis of cellulose, few have been extensively applied for process design, optimization, or control. High-solids operation of the enzymatic hydrolysis of lignocellulose is motivated by both its operation decreasing capital costs and increasing product concentration and hence separation costs. This work utilizes both insights obtained from experimental work and kinetic modeling to develop an optimization strategy for cellulose saccharification at insoluble solids levels greater than 15% (w/w), where mixing in stirred tank reactors (STRs) becomes problematic. A previously developed model for batch enzymatic hydrolysis of cellulose was modified to consider the effects of feeding in the context of fed-batch operation. By solving the set of model differential equations, a feeding profile was developed to maintain the insoluble solids concentration at a constant or manageable level throughout the course of the reaction. Using this approach, a stream of relatively concentrated solids (and cellulase enzymes) can be used to increase the final sugar concentration within the reactor without requiring the high initial levels of insoluble solids that would be required if the operation were performed in batch mode. Experimental application in bench-scale STRs using a feed stream of dilute acid-pretreated corn stover solids and cellulase enzymes resulted in similar cellulose conversion profiles to those achieved in batch shake-flask reactors where temperature control issues are mitigated. Final cellulose conversions reached approximately 80% of theoretical for fed-batch STRs fed to reach a cumulative solids level of 25% (w/w) initial insoluble solids. PMID- 18512163 TI - Automatic localization of anatomical point landmarks for brain image processing algorithms. AB - Many brain image processing algorithms require one or more well-chosen seed points because they need to be initialized close to an optimal solution. Anatomical point landmarks are useful for constructing initial conditions for these algorithms because they tend to be highly-visible and predictably-located points in brain image scans. We introduce an empirical training procedure that locates user-selected anatomical point landmarks within well-defined precisions using image data with different resolutions and MRI weightings. Our approach makes no assumptions on the structural or intensity characteristics of the images and produces results that have no tunable run-time parameters. We demonstrate the procedure using a Java GUI application (LONI ICE) to determine the MRI weighting of brain scans and to locate features in T1-weighted and T2-weighted scans. PMID- 18512164 TI - Echolucent internal carotid artery plaques are a risk factor for stroke. AB - BACKGROUND AND PURPOSE: Atherosclerotic changes in carotid arteries are responsible for 10-20% of strokes. The aim of our study was to examine how the ultrasonic morphology of carotid arteries influences the occurrence of ischaemic stroke (IS). MATERIAL AND METHODS: Ultrasonography of the carotid arteries was performed with a 7-MHz duplex-type scanner Acuson 128XP/10C. We examined 200 consecutive acute IS patients and 100 sex- and age-matched control subjects. Morphology of atheromatic plaques was evaluated with the assessment of degree of stenosis, surface regularity and echogenicity of the plaques. The predictive value of potential prognostic variables in the assessment of the risk of IS was tested using regression models. RESULTS: The most frequent site of atherosclerotic changes was the internal carotid artery (ICA) (right ICAs: 34.5% in IS group vs. 19% in controls, p=0.005; left ICAs: 26.5% vs. 16%, p=0.04). Plaques in ICAs were significantly more severe in IS patients than in controls. Echolucent plaques were observed in the IS group in 11% of right and 5.5% of left ICAs, whereas in controls we found echolucent plaques in only 2% in left ICAs. IS occurrence was independently predicted by: hypertension, congestive heart failure, current smoking status, haemodynamically significant and echolucent plaques in ICAs. CONCLUSIONS: Echolucent plaques in ICAs are an independent risk factor for ischaemic stroke even if they have no impact on haemodynamics of blood flow. Clear characterization of plaques in CAs, especially vulnerable plaques, together with estimation of the degree of stenosis, may improve the selection of patients for invasive secondary prevention methods. PMID- 18512165 TI - Efficacy of Nitric Oxide in Stroke - a randomized trial. Characteristics of patients recruited in Poland. AB - BACKGROUND AND PURPOSE: High blood pressure is frequent in patients with acute stroke. It frequently returns back to normal values within the first days after onset. It is not yet established whether it requires treatment or not. The Efficacy of Nitric Oxide in Stroke (ENOS) trial aims to assess the efficacy of transdermal glyceryl trinitrate (GTN) patches versus control and stopping or continuing antihypertensive treatments in stroke. The rationale for the study is presented and characteristics of patients recruited in Poland and elsewhere compared. MATERIAL AND METHODS: Patients were recruited to the ENOS study by 1 November, 2007. Baseline characteristics and outcome at 90 days were compared. RESULTS: Of 783 patients overall, 78 patients were recruited in Poland. The age of the patients was similar in the two groups (70.0 vs. 69.1 years). There were fewer males among Polish patients (46.8% vs. 58.3%) and fewer intracranial haemorrhages (3.8% vs. 16.7%). There were more patients with no lesions on CT (41.8% vs. 19.3%). Polish patients more frequently had hypertension (76.0% vs. 66.4%) and subsequently more frequently had antihypertensive treatment (60.8% vs. 45.3%) and nitrates (10.1% vs. 5.0%). The number of patients independent after 3 months was similar. CONCLUSIONS: Polish patients are slightly different from those treated in other countries, but at the moment the patient groups are too small to fully compare patient characteristics in different countries. The trial is ongoing and more centres are welcome to join. PMID- 18512166 TI - Results of Destandau microendoscopic lumbar discectomy. AB - BACKGROUND AND PURPOSE: Endoscopic methods are increasingly used in spine surgery, including the operative treatment of lumbar disc herniation. In this study we present a clinical series of 45 patients with lumbar disc prolapse who were operated on endoscopically with ENDOSPINE instrumentation (Karl STORZ GmbH and Co. KG), using the Destandau technique. MATERIAL AND METHODS: Between 2005 and 2006 endoscopy was used in 76 operations of lumbar disc prolapse and in 45 patients the entire procedure was performed endoscopically. All procedures were carried out from a posterior approach using a 4 mm Hopkins 0 degrees -telescope placed in the working insert equipped with channels for suction tube, operative instruments and nerve root retractor. The outcome was assessed using modified McNab criteria both directly and three months (in 35 patients) postoperatively. RESULTS: Good to excellent outcome was achieved in 89% of patients, which is consistent with results reported by experienced authors and comparable with results of "classic" microdiscectomy. Five patients reported no improvement and 3 of them were subsequently reoperated using the open surgical approach. In 3 patients the dural sac was lacerated but none of the tears exceed a few millimetres in length and they were not associated with neural injury. They were easily repaired endoscopically with a patch of "Tachocomb" tissue sealant. CONCLUSIONS: Destandau microendoscopic discectomy (MED) is a novel, safe and effective method that minimizes invasiveness of the surgical approach. Results achieved with this method are comparable to those achieved with "classic" microdiscectomy. PMID- 18512167 TI - Accessory deep peroneal nerve - a clinically significant anomaly? AB - BACKGROUND AND PURPOSE: The accessory deep peroneal nerve (ADPN), a variant of the peroneal nerve, may give motor branches to the extensor digitorum brevis muscle (EDB) in 19-28% of the general population and in up to 78% of subjects in familial cases. The aim of our study was to evaluate its significance in the examination of the peroneal nerve. MATERIAL AND METHODS: Three groups of patients were analyzed. Group I consisted of 310 patients whose neurography recordings were analyzed retrospectively. Group II consisted of 24 healthy controls and group III consisted of 8 relatives of a healthy control having the ADPN detected. All patients underwent routine neurography of the peroneal nerve with muscular response recorded from the EDB. Groups II and III had additional stimulation behind the lateral malleolus. RESULTS: On routine neurography, ADPN was found in 7.7% of patients in group I and 12.5% in group II. Stimulation behind the lateral malleolus detected it in 25% of group II and in 50% of group III. The highest CMAP amplitude obtained by stimulation of the ADPN equalled 3.71 mV and was over half of the total EDB response. The presence of the ADPN significantly influences routine neurography of the peroneal nerve in 7.7-12.5% of patients. Stimulation behind the lateral malleolus detected it in 25% of group II and in 50% of the maternal line of the family in group III. In the familial case ADPN was present in 50% of maternally related subjects, reflecting autosomal dominant transmission. CONCLUSIONS: ADPN may innervate the greater part of the EDB and in cases of peroneal neuropathy may be important for preserving function of the muscle. PMID- 18512168 TI - Results of rehabilitation in patients after subarachnoid haemorrhage from ruptured intracranial aneurysm and after surgical treatment. AB - BACKGROUND AND PURPOSE: Non-traumatic, or spontaneous subarachnoid haemorrhage (SAH) is responsible for 5-7% of strokes. In Podkarpackie voivodeship, 217 patients were hospitalized in 2003 with spontaneous SAH. The aim of this study was to assess the relationship between the results of rehabilitation carried out in patients after surgical treatment of SAH estimated on the Brunnstrom, Barthel, and Rankin scales and patients' initial condition measured by means of the Hunt Hess Scale. MATERIAL AND METHODS: The examined group consisted of 55 patients after surgically treated SAH (clipping). Severity of bleeding was graded with the Hunt-Hess Scale on admission to the neurosurgery ward. Patients' clinical status was assessed with the Barthel Index and Rankin and Brunnstrom scales on admission to a rehabilitation ward, and rehabilitation effects were assessed after 4 weeks of the rehabilitation period and after at least 6 months from the end of rehabilitation on the ward. RESULTS: The study did not reveal any significant correlation between severity of bleeding and the results of rehabilitation. No explicit advantage was observed in the process of treatment of patients with grade I or II grade according to the Hunt-Hess Scale in the later period. In each group of patients (grades I to IV) there were patients whose final condition improved evidently. CONCLUSIONS: No significant correlation was observed between the clinical status at the onset of the disease and clinical or functional status in early and late periods of rehabilitation. PMID- 18512169 TI - [CADASIL syndrome - cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy]. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited neurodegenerative disease associated with mutations in the Notch 3 receptor on vascular smooth muscle cells. Clinically the syndrome is manifested as migraine, recurrent subcortical ischaemic events, dementia and mood disorders. CADASIL, considered one of the important causes of "subcortical vascular dementia", is relatively easy to overlook or misdiagnose if it is not taken into consideration in differential diagnosis. Diagnosis of CADASIL is established on the basis of results of skin biopsy and genetic examination. In this article we present a short review of the literature concerning pathogenesis and clinical presentation of the syndrome and provide recommendations for detection, diagnosis and management strategies. PMID- 18512170 TI - [Pharmacogenetics in Parkinson's disease treatment]. AB - The authors present the current opinion on the significance of molecular biology in individualized therapy. Pharmacogenetics is a new branch of clinical pharmacology dealing with the influence of genetic factors on drugs with special focus on interpersonal differences to drug response. The article includes basic rules of pharmacogenetics as well as its use in clinical practice. Individualized treatment of Parkinson's disease is not widely known although interpersonal differences to drug response is clearly stated. There is some evidence that varied efficacy of treatment and risk of motor and mental complications can be of genetic origin. Some results concerning the relationship between genetic polymorphism of COMT, DRD2, DAT, CCK, MTHFR and successful and safe treatment of Parkinson's disease are presented. PMID- 18512171 TI - [Posturography and computerized gait analysis in the Computer Dyno Graphy system as non-invasive methods for evaluation of normal pressure hydrocephalus progression]. AB - Disturbances of gait and balance are important clinical problems in patients with normal pressure hydrocephalus. A considerable degree of locomotor disability and increased risk of falls are present in many cases. Accordingly, attempts to develop effective evaluation methods of gait and balance disturbances are made. Posturography and the Computer Dyno Graphy (CDG) system appear to be useful in diagnosis and evaluation of treatment in patients with normal pressure hydrocephalus. Both are non-invasive and repeatable methods that enable accurate evaluation of analyzed parameters at time intervals. PMID- 18512172 TI - Cerebral abscess caused by Nocardia nova. AB - Primary cerebral nocardiosis is very rare and the subgroup of the isolate has usually been defined as Nocardia asteroides. We report an unusual patient with a cerebral abscess whose cultures were typical for N. nova. It was first isolated from an HIV-1 infected individual and has a distinctive antibiotic susceptibility among the species of N. asteroides. Aggressive surgical and medical intervention is a must for these unusual pathogens. Although our surgical strategy was successful, treatment of nocardial cerebral abscess is quite hard and long lasting, and N. nova in particular necessitates a specific antibiotic regimen. PMID- 18512173 TI - Acute stroke-elicited epilepsia partialis continua responsive to intravenous sodium valproate. AB - We present an interesting case of epilepsia partialis continua (EPC) responsive to valproate treatment as an acute manifestation of cortical ischaemia in the course of vasospasm after subarachnoid bleeding A 72-year-old, right-handed woman suffering a subarachnoid haemorrhage complicated with intracerebral left hemispheric haematoma was admitted to our Department of Neurology In the disease course acute vasospasm developed, affecting the contralateral hemisphere Epilepsia partialis continua (EPC) ensued as an acute complication, with clonic jerks of the left side of the face and left upper limb This was related to epileptiform ictal activity which was followed by periodic lateralized epileptiform discharges (PLEDs) from the right frontal lobe When the standard treatment with benzodiazepines and phenytoin failed, the infusion of sodium valproate (VPA) was started This completely abolished seizure activity, bringing an improvement of the patient's neurological condition EPC may be an acute complication of cortical ischaemic damage and valproate intravenous preparation is an interesting alternative for the treatment of this poorly studied condition. PMID- 18512174 TI - Tumour-like lesions in multiple sclerosis. AB - The presence of tumefactive lesions on magnetic resonance imaging (MRI) in multiple sclerosis (MS) patients can cause diagnostic difficulties. It requires differential diagnosis between tumefactive demyelinating lesion (TDL) and the coexistence of neoplasm; it also implies further management. The precise assessment of such lesions at the first clinical manifestation of the disease is particularly important. We present three cases of MS presenting with tumour-like lesions of the brain. Based on serial MRI studies, stereotactic biopsy and the response to treatment with corticosteroids, the diagnosis of TDL was established in every case. PMID- 18512175 TI - [Neurofibromatosis type 2 in a child - diagnostic difficulties]. PMID- 18512176 TI - Validity and reliability of the Ergomopro powermeter. AB - The aim of this investigation was to assess the validity and reliability of the Ergomopro powermeter. Nine participants completed trials on a Monark ergometer fitted with Ergomopro and SRM powermeters simultaneously recording power output. Each participant completed multiple trials at power outputs ranging from 50 to 450 W. The work stages recorded were 60 s in duration and were repeated three times. Participants also completed a single trial on a cycle ergometer designed to assess bilateral contributions to work output (Lode Excaliber Sport PFM). The power output during the trials was significantly different between all three systems, (p < 0.01) 231.2 +/- 114.2 W, 233.0 +/- 112.4 W, 227.8 +/- 108.8 W for the Monark, SRM and Ergomopro system, respectively. When the bilateral contributions were factored into the analysis, there were no significant differences between the powermeters (p = 0.58). The reliability of the Ergomopro system (CV%) was 2.31 % (95 % CI 2.13 - 2.52 %) compared to 1.59 % (95 % CI 1.47 to 1.74 %) for the Monark, and 1.37 % (95 % CI 1.26 - 1.50 %) for the SRM powermeter. These results indicate that the Ergomopro system has acceptable accuracy under these conditions. However, based on the reliability data, the increased variability of the Ergomopro system and bilateral balance issues have to be considered when using this device. PMID- 18512177 TI - Validation of a soccer skill test for use with females. AB - The aim of this study was to validate the Loughborough Soccer Passing Test (LSPT) for use with female players. Nineteen Premier Division (elite) and 16 Reserve team players (nonelite) volunteered for this study. The LSPT requires players to complete 16 passes against coloured target areas as fast and as accurately as possible. Participants completed an initial familiarisation trial followed by two main trials, each separated by at least one week. During both trials participants were given two practice efforts before recording the mean of the next two attempts as the performance score. All trials were performed inside a sports hall, using an indoor soccer ball, and following a standardised 10-min warm-up. The mean time taken (54.6 +/- 5.3 s vs. 61.6 +/- 6.5 s, p = 0.002), added penalty time (22.8 +/- 7.2 s vs. 35.9 +/- 11.5 s, p < 0.001) and overall performance time (77.4 +/- 11.6 s vs. 97.5 +/- 17.2 s, p < 0.001) were lower for elite players. Due to the lower agreement ratio, LSPT performance was more repeatable in the elite (x//1.39) relative to nonelite (x//1.45) group. In conclusion, the LSPT is a valid and reliable protocol to assess differences in soccer skill performance using female players. PMID- 18512178 TI - Air pollution and sports performance in Beijing. AB - The Beijing Olympics will begin in August 2008 and athletes will face an unpredictable challenge. Based on present data, Beijing is one of the most polluted megacities in the world; the air concentrations of carbon monoxide (CO), ozone, nitrogen oxides (NO and NO2), sulphur dioxide (SO2) and particulate matter approach or exceed the current limits established by U. S. Environmental Protection Agency (EPA). Although the athletes who will be competing in Beijing are physiologically very different to the participants in most published studies, and it is therefore difficult to predict individual responses, there is little doubt that the presence of these air pollutants might be detrimental to athletic performance due to the marked increase (up to 20-fold) in ventilatory rate and concomitant nasal and oral breathing. Moreover, mouth breathing often bypasses the noise during strenuous exercise, increasing the deleterious effects of pollutants on health and athletic performance. Although limited, each decrement in athletic performance would have a potentially deleterious impact on top-class athletes competing in the next Olympics in China. Several Olympic records are regularly broken during the Olympics. Will this be the case for Beijing? PMID- 18512179 TI - The effect of a 9-week physical activity programme on bone and body composition of children aged 10-11 years: an exploratory trial. AB - A high-impact exercise and a lifestyle intervention were implemented over a 9 week period; changes in bone and body composition were compared to controls. Sixty-one children volunteered from three randomly selected schools. Each school was randomly assigned to either a structured exercise (STEX) intervention, a lifestyle intervention (PASS) or control (CONT). Bone mineral content (BMC) and density (BMD) of total body, femoral neck and lumbar spine were measured as well as fat and lean mass at baseline and post-intervention by dual-energy X-ray absorptiometry. The STEX intervention resulted in an additional mean increase in total body BMC of 63.3 g (p = 0.019) and an additional increase of 0.011 g . cm ( 2) (p = 0.018) for BMD over increases observed by controls. Bone mineral increases observed for the PASS intervention were not significant compared to the control group (p > 0.05). Neither intervention produced significant increases in bone mineral at femoral neck or lumbar spine sites (p > 0.05) compared with the controls. No significant changes were found in fat mass index (p > 0.05), lean mass index (p > 0.05) or percent body fat (p = 0.09) in any groups. Structured impact exercise promoted significant and clinically relevant increases in bone measures, without significant changes to body composition. A larger, definitive randomised trial is needed to confirm the present results. PMID- 18512180 TI - Fluid tolerance while running: effect of repeated trials. AB - This study assessed tolerance to fluid ingestion with repeated sessions of drinking while running. Runners (n = 7; age 22 +/- 2 yr; V O (2max) = 54.4 +/- 7.1 ml/kg/min) performed six 90-min runs (65 % V O (2max); separated by 7 - 11 days). During run 1, subjects drank a glucose-electrolyte solution AD LIBITUM for 1 min every 10 min. During runs 2 - 6, subjects drank a volume of the solution every 10 min equal to their sweat production over 10 min during run 1. Stomach comfort (1 - 4 scale) and gastrointestinal symptoms were also assessed every 10 min. Gastric emptying rate was determined in runs 2 and 6. Subjects consumed more (p < 0.05) fluid during runs 2 - 6 (mean +/- SD; 1247 +/- 162 ml), than during run 1 (508 +/- 476 ml). Stomach comfort improved (p < 0.05) on runs 5 and 6 (1.7 +/- 0.5 mean ranks) compared to run 2 (2.3 +/- 0.5 mean ranks). Gastric emptying rate was not different between runs 2 and 6 (12.0 +/- 1.9 ml/min vs. 12.3 +/- 2.3 ml/min, respectively). These results indicate repeated sessions of drinking at a rate matching sweat rate improves stomach comfort, however, gastric emptying rate does not change under such conditions. PMID- 18512181 TI - Long-term follow-up of bradycardia in elite athletes. AB - The aim of the study was to examine the hypothesis that when former high-level athletes retired from competition, bradycardia would vanish. ECG changes and factors possibly related to the persistence of bradycardia were investigated. We performed a longitudinal follow-up study in 157 former elite athletes who had records of bradycardia < 50 bpm when they were active in high-level competition. All had retired from competitive sport for a minimum of five years prior to participation in the follow-up examination. Data on sport modality, hours of weekly training during competition periods, years participating in high-level competition, and years since retirement from competition were abstracted from medical records at two sports medicine centers. ECGs from this time were studied. At the post-retirement follow-up examination, participants underwent a clinical examination that included blood pressure and ECG recordings, and answered a structured questionnaire about their physical exercise since retirement from their sporting career. In the post-retirement period, a total of 65 % of participants had persistent bradycardia, 18 % with bradycardia < 50 bpm. Multivariate analysis showed that persistence of resting bradycardia was associated with regular exercise and number of years in high-level competition, but not with symptoms such as palpitations, dizziness or syncope, or major ECG alterations. PMID- 18512182 TI - [How progressive is the journal "Fortschritte Neurologie. Psychiatrie"?--a two years balance]. PMID- 18512183 TI - [Changing care for the mentally ill in the "house of the poor insane in Lubeck" from the 17th to the early 19th century]. AB - Until quite recently the history of the "House of the Poor Insane" in the Hanseatic City of Lubeck has only been studied selectively. Our study comprises an overall view of the comprehensive historical documents concerning this institution during the 17th and 18th century; the period of the following 40 years in another new constructed house is outlined likewise. Not only the journal, into which the principals of the institution wrote down the minutes, but foremost the cassa-books without omissions were used to describe the social history as well as the range of activities. Thus historical windows into the daily routine of the inmates are opened. - During the greater part of 17th century, the institution rather resembled a prison for the mentally ill, though in the end of the 17th and during the 18th century an important early phase of reform is to be recognized, followed, however, by a period of repression and "containment" towards the end of the century and in the beginning of the 19th century. Then again, this time initiated by the medical profession, the reform was revived. - The Lubeck findings are not only compared with the historical development of inpatient treatment in institutions elsewhere, but it is also discussed, which influences were the decisive factors for the public care in the city of Lubeck. PMID- 18512184 TI - [Hallucinogen-induced psychological disorders]. AB - OBJECTIVE: The purpose of this article is to provide an overview of the current research on hallucinogen induced psychiatric disorders. In addition to LSD and psilocybin hallucinogens of biologic origin are increasingly used by adolescents and young adults. METHODS: Relevant literature and related articles were identified by means of a computerized MEDLINE search including the years 1997 - 2007. As keywords "hallucinogen induced psychosis", "hallucinogen induced flashback", "hallucinogen persisting perception disorder (HPPD)" were used. Finally, 64 journal articles and books out of 103 were included in the review. RESULTS: Acute psychotic syndromes in adolescents are rarely due to intoxications with hallucinogenic drugs. However, clinical relevance of flashback phenomena as post-hallucinogenic psychiatric disorder has to be disputed. Because of the high popularity of biogenic hallucinogens and LSD knowledge of intoxications and resulting psychiatric disorders as well as medical complications and therapeutical approaches are clinically important. Especially intoxications with drugs of herbal origin like tropanalcaloids play an important role in emergency situations. PMID- 18512185 TI - [Criminal responsibility in sexual homicide perpetrators]. AB - OBJECTIVE: Evaluation of criteria for criminal responsibility, esp. in paraphilias and personality disorders. METHODS: We compared sexual murderers with diminished responsibility or not-guilty for reasons of insanity (n = 73) and those held fully responsible (n = 61) regarding psychiatric disorders, signs of severe paraphilias, level of personality structure (using Operationalized Psychodynamic Diagnostics) and offence related criteria. RESULTS: Sexual murderers found as diminished responsible or not-guilty, especially those detained in forensic psychiatric hospitals, showed more signs of a progressive sadistic development and lower levels of personality structure. Offence related criteria that are regarded as not supporting a diminished capacity plea (purposeful actions, long duration, complex, stepwise course of the offence) were found more frequent in those offenders that were detained in forensic hospitals. CONCLUSIONS: Signs for severity of paraphilias and personality disorders are useful in decisions about criminal responsibility. Criteria regarding purposeful course of action appear not to be adequate for the assessment of paraphilic sexual offenders. PMID- 18512186 TI - [Modern neurophysiological strategies in the rehabilitation of impaired hand function following stroke]. AB - Modern neurophysiological brain stimulation techniques, such as transcranial magnetic stimulation and direct current stimulation, are powerful tools to inhibit or facilitate cortical excitability for several minutes after stimulation depending on the stimulation parameters used. Purposeful modulation of cortical excitability may induce plastic changes within the cortical network of sensorimotor areas, and has the power to improve the function of the affected hand after stroke. The therapeutic use of transcranial brain stimulation techniques is based on the concept of interhemispheric competition. Here we give an overview of the use of repetitive transcranial magnetic stimulation and direct current stimulation in the rehabilitation of impaired hand function after stroke. PMID- 18512187 TI - [Parainfectious polyneuropathy and miller-fisher-syndrome in combination with anemia in Mycoplasma pneumoniae infection]. AB - Neurological complications are among the most common in Mycoplasma pneumoniae (M.p.) infection and appear in up to 4,8 % of M.p.-infected patients . Whereas direct intracerebral infection can occur and often puts patients into critical, intensive care-requiring conditions, different forms of parainfectious autoimmune Guillain-Barre-syndrome (GBS) and Miller-Fisher syndrome (MFS) make up the most frequent neurological complications. M.p.-associated alterations of the blood count are common, and sometimes can take a dramatical course. We here report a 64 year-old female and a 53-year-old male patient who both suffered from severe parainfectious autoimmune neurological syndromes in combination with different forms of anemia, in the case of the first patient to a transfusion-requiring degree. With this report we would like to stress the importance to include M.p. in the diagnostic procedures in the case of inflammatory neurological syndromes, especially when combined with alterations of the blood count. PMID- 18512188 TI - [Road safety under psychiatric drug targeting]. PMID- 18512189 TI - [Cryptogenic organizing pneumonia: typical and atypical imaging features on computed tomography]. AB - Organizing pneumonia (OP) occurs without any identifiable cause ("cryptogenic organizing pneumonia") as well as secondary to a multitude of disorders of various origins ("secondary organizing pneumonia"). Possible triggers are infections, drugs, collagen vascular disease, inflammatory bowel disease, transplantations, and radiation directed to the chest. The present manuscript provides an overview of the histopathological, clinical and CT imaging features of OP. Classic CT morphologies (peripheral and peribronchovascular consolidations and ground glass opacities) and atypical imaging features (nodules, crazy paving, lines and bands, perilobular consolidations and the reversed halo sign) are discussed. PMID- 18512190 TI - [Quantitation of glutamate in the brain by using MR proton spectroscopy at 1.5 T and 3 T]. AB - PURPOSE: The influence of different magnetic field strengths on the quantification of glutamate was experimentally investigated by means of IN VITRO and IN VIVO (1)H-MR spectroscopic measurements at 1.5 T and 3 T. MATERIALS AND METHODS: In vitro (1)H-MR measurements of aqueous solutions of NAA, glutamate, glutamine and GABA were performed on two clinical MR scanners at 1.5 T and 3 T using a single voxel PRESS sequence (TR/TE = 10 000 / 30 ms). IN VITRO brain measurements were also performed at both field strengths using a PRESS 2D- (1)H CSI-sequence (TR/TE = 5000 / 30 ms) in 6 volunteers. Spectra at 1.5 T and 3 T were compared with respect to the overlap of the single compound spectra and the deviations between estimated and nominally adjusted concentrations. In vivo spectra at both field strengths were compared with respect to SNR (Glu), line width and Cramer-Rao values of the estimated glutamate intensities by using the LCModel. For the thalamus, insular and parietal cortex mean Glu/tCr ratios were estimated and compared between 1.5 T and 3 T as well as with corresponding values in the literature. RESULTS: In general, an improved separation of signal maxima was observed in the IN VITRO spectra at 3 T. Except for GABA, all IN VITRO concentrations estimated at 3 T revealed lower deviations from their adjusted nominal concentration compared to 1.5 T: NAA (1.5 T: -5.5 %, 3 T: 0.7 %), glutamate (1.5 T: -18.1 %, 3 T: 12.3 %), glutamine (1.5 T: 44.8 %, 3 T: 9.2 %), GABA (1.5 T: - 24.8 %, 3 T: 33.8 %). The SNR of IN VIVO spectra at 3 T was nearly doubled compared to 1.5 T. The mean number of voxels with %SD (Glu)< 20 was distinctly lower at 1.5 T (53 %) than at 3 T (80 %). Estimated Glu/tCr ratios for thalamus, insular and parietal cortex lay in the upper range of the literature values. CONCLUSION: The results indicate that the advantageous distribution of signal maxima at 3 T allows an improved separation of the individual spectra. Both the higher initial magnetization at 3 T and the improved sensitivity of the phased array matrix coil used in the 3 T study result in an increased SNR, which leads to better reliability of the individual detection as well as a more accurate quantification of glutamate. PMID- 18512191 TI - [Do hospital physicians really want to go digital? --Acceptance of a picture archiving and communication system in a university hospital]. AB - PURPOSE: Radiology departments are making the transition from analog film to digital images by means of PACS (Picture Archiving and Communication System). It is critical for the hospital that its physicians adopt and accept the new digital work method regarding radiological information. The aim of this study is to investigate hospital physicians' acceptance of PACS using questionnaires pre- and post-implementation and to identify main influencing factors. MATERIALS AND METHODS: The study was conducted in an 1169 bed university hospital. The UTAUT (Unified Theory of Acceptance and Use of Technology) questionnaire was administered at two times: one month pre-implementation (T1) and 1.5 years post implementation (T2) of PACS, targeting all hospital physicians with the exemption of radiologists. The UTAUT scales (Behavioral Intention BI; Facilitating Conditions FC; Effort Expectancy EE; Performance Expectancy PE; Anxiety ANX; Social Influence SI; System Use USE; Attitude toward technology ATT; Self Efficacy SE) were used to assess questions regarding: a) PACS' usefulness, b) PACS' ease of learning/using, c) PACS support availability, d) the perceived pressure to use PACS, e) physicians' attitude towards PACS and f) physicians' intention to use and actual use of PACS. RESULTS: At T 1 scale ratings were positive toward the PACS implementation. The ratings on all scales with the exception of self-efficacy improved at T 2. Regression analysis revealed that the key factor for intention to use PACS at T 1 was the usefulness of PACS, while the availability and awareness of support was its most important predictor at T 2. Overall, PE was the best predictor of BI, but all four UTAUT-determinants (PE, FC, EE and SI) were salient for its prediction. Variance explained in BI ranged from 31 to 37 % while variance explained in USE was very low (3 %). CONCLUSION: The implementation of PACS has succeeded. At T 1 the physicians were welcoming PACS and this was confirmed at T 2. Experience with PACS led to an overall improved attitude toward PACS. The key factors for physicians' intentions to use PACS were the usefulness of PACS (at T 1 and overall) and the availability of support (at T 2). PMID- 18512192 TI - [Detection of bone metastasis of prostate cancer - comparison of whole-body MRI and bone scintigraphy]. AB - PURPOSE: Prostate cancer continues to be the third leading cancer-related mortality of western men. Early diagnosis of bone metastasis is important for the therapy regime and for assessing the prognosis. The standard method is bone scintigraphy. Whole-body MRI proved to be more sensitive for early detection of skeletal metastasis. However, studies of homogenous tumor entities are not available. The aim of the study was to compare bone scintigraphy and whole-body MRI regarding the detection of bone metastasis of prostate cancer. MATERIALS AND METHODS: 14 patients with histologically confirmed prostate cancer and a bone scintigraphy as well as whole-body MRI within one month were included. The mean age was 68 years. Scintigraphy was performed using the planar whole-body technique (ventral and dorsal projections). Suspect areas were enlarged. Whole body MRI was conducted using native T 1w and STIR sequences in the coronary plane of the whole body, sagittal imaging of spine and breath-hold STIR and T 1w-Flash 2D sequences of ribs and chest. Bone scintigraphy and whole-body MRI were evaluated retrospectively by experienced radiologists in a consensus reading on a lesion-based level. RESULTS: Whole-body MRI detected significantly more bone metastasis (p = 0.024). 96.4 % of the demonstrated skeletal metastases in bone scintigraphy were founded in whole-body MRI while only 58.6 % of the depicted metastases in MRI were able to be located in scintigraphy. There was no significant difference regarding bone metastasis greater than one centimeter (p = 0.082) in contrast to metastasis less than one centimeter (p = 0.035). Small osteoblastic metastases showed a considerably higher contrast in T 1w sequences than in STIR imaging. Further advantages of whole-body MRI were additional information about extra-osseous tumor infiltration and their complications, for example stenosis of spinal canal or vertebral body fractures, found in 42.9 % of patients. CONCLUSION: Whole-body MRI using native STIR and T 1w sequences is superior to bone scintigraphy for the detection of small bone metastasis of prostate cancer. Simultaneous clarification of associated complications demonstrates further advantages. PMID- 18512193 TI - [Impact of technical and morphological factors on the precision of software-based MR tumor volumetry: a phantom study]. AB - PURPOSE: To investigate the impact of technical and morphological parameters on the precision of software-based MR tumor volumetry (SBV) in an in-vitro experimental setting. MATERIALS AND METHODS: Tumor models were formed from a silicone compound in three different sizes with a max. diameter < 2 cm (small), 2 - 4 cm (middle), and > 4 cm (large). For each size a spherical, an elliptic and an irregular shaped model was produced. The true volume of the tumor models was established by water displacement. Tumor models were examined with a high-field MRI (TRIO, 3 Tesla, Siemens) with T 2-weighted sequences under optimized contrast conditions. Slice thickness was 1, 3 and 5 mm. The volume of the tumor models was then calculated using (1). manually driven volumetric software (SBVmanual) and (2). automatic volumetric software (SBVauto). The influence of the following parameters on the precision of SBV was analyzed: Size and shape of the tumor models, manual/automatic SBV, segmentation technique and slice thickness. RESULTS: In general, SBVauto measurements showed less deviation than measurements with SBVmanual (p < 0.01). However, both methods depended significantly on morphologic factors, especially on tumor size. In small tumor models, the volume was strongly underestimated by -36.2 +/- 27.8 % (SBVmanual) and -33.1 +/- 8.6 % (SBVauto), whereas the deviation for large tumor models was only 2.0 +/- 14.7 % (SBVmanual) and 3.0 +/- 2.3 % (SBVauto; p < 0.01). The deviation of measurements increased from the "spherical" to the "irregular" shape by 9.5 % (SBVmanual) and 10.7 % (SBVauto). In addition, SBVmanual depended on technical factors. Using a "minimal" segmentation technique (e. g. excluding partial volume effects), volumes were underestimated in all cases, whereas volumes of middle and large tumor models were slightly overestimated when using a "maximum" segmentation technique (e. g. including partial volume effects; p = 0.01). Deviation of SBVmanual increased with slice thickness from 15.9 +/- 12.7 % (1 mm slices) to 27.1 +/- 21.3 % (5 mm-slices). CONCLUSION: In general, SBVauto measurements yielded smaller deviations than SBVmanual. However, both methods showed major inaccuracy in the volumetric estimation of small and irregular shaped tumor models, thus the tumor volumetry of these tumors has to be considered inappropriate for clinical practice. Moreover, the exactness of SBVmanual depended significantly on segmentation technique and slice thickness. PMID- 18512194 TI - [Pseudotumor of the eye--idiopathic orbital inflammatory diseased (IOID)--a case report of an unusual manifestation of a common disease]. PMID- 18512195 TI - [Minimum hospital volumes for total knee replacement]. AB - BACKGROUND: As requested by the Federal Joint Committee, the German Hospital Institute and the Heinrich-Heine University of Dusseldorf carried out an investigation of the minimum volume regulation for hospitals based on the Social Legislation Code. Total knee replacement forms one minimum volume field. Since 2006 hospitals with a performance rate of less than the minimum volume of 50 patients a year with knee replacement are no longer permitted to conduct this procedure. The object of the present analysis is to investigate the impact of the minimum volume regulation for total knee replacement. METHODS: The results are based on two hospital surveys on the application of the minimum volume regulation for total knee replacement. 279 hospitals (response rate: 41,8%) participated in 2006 and 297 hospitals in 2007 (response rate: 47,5%). The results are representative of General hospitals with total knee replacements. RESULTS: As expected, hospitals above and below the minimum volume cut-off differ in size. To date the minimum volume regulation has led to a rather selective exclusion of hospitals from care. In the case of total knee replacement 13,7% of the hospitals have been excluded. Most hospitals that do not reach the minimum volume are still participating in care. A decisive reason for this is the existence of exception rules. In hospitals exceeding the minimum volume, certain quality management tools for knee replacement are more widely spread than in hospitals that do not reach the minimum volume. As a consequence of the minimum volume regulation, the participating hospitals improved their position in the market. Vice versa, the excluded hospitals are more concerned about the damage to their image that may result from being excluded from care. With respect to the further development of the minimum volume regulation, the hospitals do not share the same point of view. DISCUSSION: Because, as yet, only few hospitals with low case numbers have been excluded from care, the immediate effects of the minimum volume regulation on the affected hospitals and hospital care in general are limited. The surveys showed a considerable uncertainty among all participants about the application and effects of the minimum volume regulation in hospitals. PMID- 18512196 TI - [The economic benefits of self-help associations]. AB - OBJECTIVES: This article primarily reflects on the economic benefits of self-help associations, and the problems of their adequate evaluation. METHODS: We present a critical discussion of the concepts in the economic evaluation and empiricism by secondary analysis of data available based on a review of published studies as well as the re-analysis of primary data from our Hamburg study in 2003 (n=345 or 52.4% of the self-help associations registered in Hamburg). RESULTS: Most approaches to evaluate the economic benefits of self-help groups were inspired by the idea of output-related, cost-benefit analysis, by which activities of members were monetarily estimated. Projected for Hamburg, the productive performance amounts to nearly 1.8 million Euro per month, and 21.5 million per year. Furthermore, additional voluntary commitment can be mobilized at 300,000 Euro per month. CONCLUSION: All calculations are based on some assumptions (e.g., coefficient of productivity, wages) which cannot be finally verified. In view of the state of research reviewed we strongly recommend to bring forward both outcome-oriented approaches of economic evaluation, and new representative studies of self-help groups for Germany as a whole. PMID- 18512197 TI - [Particulate matter in classrooms--problem and the impact of cleaning and ventilation with the City of Frankfurt am Main as an example]. AB - BACKGROUND: Indoor air quality in schools, especially the levels of particulate matter in classrooms, have become a matter of great public concern in Germany, Convenient recommendations to improve the hygienic situation in classrooms via cleaning and ventilation are often not followed. Here, data on particulate matter in the indoor air of classrooms, before and after intensified cleaning, are reported. MATERIAL AND METHODS: In the winter of 2006, analyses for particulate matter were done in two primary schools in Frankfurt/M, one of them located in a rural area the other one in an inner city area of Frankfurt/M. Particulate matter (PM10) was analysed by laser beam technology, the data documented every five minutes from 8 am to 2 pm. In parallel, a documentation of the number of persons present in the room, their activity and ventilation was done according to a standardised protocol. Measurements were collected for a period of three weeks. During the first week, the normal school situation was analysed, i.e., the classrooms were ventilated as usual and were cleaned by wet wiping twice a week. During the second and third weeks, the rooms were cleaned every day, so that the effect of intensified cleaning could be studied. In winter 2007, these measurements were continued for one week in two classrooms of a passive house school, following the identical study protocol. RESULTS: During the first week- wet cleaning twice a week--mean particulate concentrations of 86 microg/m3 (median 60 microg/m3) were obtained, during the 2nd and 3rd weeks--wet cleaning every school day mean concentrations of 60 microg/m3 (median 53 microg/m3)--with great differences on different days. Especially during and after handicraft (casting candles) lessons, maximum levels up to >1000 microg/m3 were analysed. Although an impact of cleaning on levels of indoor particles could be established, indoor PM10 levels were dominated by indoor factors, such as occupancy and activity of the persons in the room. After substraction of the outdoor PM10 levels from the indoor levels, the "indoor part" was ca. 50 microg/m3 before, and ca. 30 microg/m3 during intensified cleaning. Further detailed investigations showed the predominance of particles >1 microm indoors, which could easily be diminished by cleaning and ventilation. Indoor particles <0.5 microm, however, were increased via ventilation. CONCLUSION: In classrooms, generally higher indoor levels of air particulate matter are detected than outdoors. Particulate matter in the indoor air of classrooms is to be considered as an indicator of low hygiene and of increased and avoidable heath risk for pupils and teachers. Our data indicate the relevance of cleaning--and with regard to PM <1 microm also of ventilation--for the reduction of particulate matter in classrooms. Therefore, these measures should be taken to improve indoor air quality in schools. PMID- 18512198 TI - [School-based smoking prevention by enhancement of life competence--an empirical evaluation]. AB - AIM OF THE STUDY: An intervention for the prevention of smoking in fifth grades of basic secondary schools is to be evaluated. METHOD: A non-randomised control group study was performed. The intervention-group consisted of 21 classes from 9 secondary schools of the County of Neuss, the control group of 34 classes from 12 schools of the counties of Neuss and Erft. The intervention consisted of up to 12 teaching units in one academic year with the goal of giving smoking-relevant information and of strengthening social competences. Before and after the intervention, pupils in both study groups answered questionnaires anonymously. The questions focussed on smoking behaviour in the previous four weeks (smoking frequency) and, limited to pupils who had never smoked before in the first interview (never-before-smokers), on the start of smoking. RESULTS: A total of 1,082 pupils participated in both interviews (intervention: 462; control: 620). Smoking frequency increased more within the intervention group than in the control group (p < 0.05). In the intervention group more never-before-smokers started smoking than in the control group (12.7% versus 7.8%; p < 0.05). After statistical control of age, sex, nationality, smoking status of relatives and the interaction of these variables with the intervention, both effects are no longer statistically significant. Instead another result is emerging: for pupils with a smoking father, smoking frequency in the intervention group increases significantly less than in the control group (p<0.01). CONCLUSIONS: The intervention had no general effect for all pupils. However, it did have a positive effect specifically for pupils with a smoking father. The whole intervention does not seem to be cost-effective. Specific interventions with a focus on endangered groups could be more cost-effective. PMID- 18512199 TI - [Gender-specific differences in quality of life of patients with type 2 diabetes with or without participating in DMP]. AB - OBJECTIVE: Disease management programmes (DMP) are supposed to improve the care of patients with type 2 diabetes or other chronic conditions. One stated aim is the improvement of the health-related quality of life. Within the ELSID study (controlled study for the evaluation of the DMP for patients with type 2 diabetes) there has been a survey of insurants of the general regional health funds (AOK) by means of the SF-36. The aim of this survey is a comparison of patients participating in the Diabetes DMP with those who are not participating in the program with regard to their quality of life. METHODS: A random sample of 3,546 patients with type 2 diabetes out of the total sample of the ELSID study (20,625) was asked to complete the SF-36. RESULTS: 1,532 questionnaires were returned (response rate 43.2%). 1,399 were analysed. Within all scales of the SF 36, men achieved higher scores than women. Differentiated in participants and non participants in the DMP, in men there were lower scores for the participants than for the non-participants. In women this proportion was reversed. CONCLUSIONS: This cross-sectional-study provides first indications for gender-specific differences within the quality of life of patients with diabetes participating in the DMP compared to patients who are not participating. These differences should be considered more intensely within further research of DMP evaluation and the configuration of the programmes in the future. PMID- 18512200 TI - [Imported malaria in Germany--countries of infection and parasite species, 1993 2007]. AB - From 1993 to 2007, a total of 11,605 malaria cases were reported to the Robert Koch-Institut. For 9 593 of these cases information on the possible country of infection is available. Analysis of the data indicates that the malaria cases were imported to Germany from 96 countries. The majority of the malaria cases were acquired in Africa, followed by distinctly fewer cases from Asian countries. More than half of all cases were imported from only 6 countries (Ghana, Kenya, Nigeria, Cameroon, Gambia, and India). The number of cases imported from, e.g., Kenya and India has declined, whereas infections from Ghana, Nigeria, and Cameroon have increased during the last years. For more than two-thirds of all infections the predominant parasite species was P. falciparum; this is consistent with the predominance of the African continent as the main source of infection. PMID- 18512202 TI - Nerve grafts prevent paraplegic pressure ulcers. AB - We present an optimistic neural solution to the depressing challenge of decubitus pressure ulcers in the paraplegic patient. This is a limited study of two paraplegic men followed for several years. Sural nerve grafts, performed end-to side, successfully bridged the sciatic nerve to intercostal nerves with surprising benefits for both men. PMID- 18512201 TI - Fetal heart rate monitoring patterns in women with amniotic fluid proteomic profiles indicative of inflammation. AB - We hypothesized that abnormal fetal heart rate monitoring patterns (FHR-MPs) occur more often in pregnancies complicated by intra-amniotic inflammation. Therefore, our objective was to examine the relationships among FHR-MP abnormalities, intra-amniotic inflammation and/or infection, acute histological chorioamnionitis, and early-onset neonatal sepsis (EONS) in pregnancies complicated by preterm birth. Additionally, the ability of various FHR-MPs to predict EONS was investigated. FHR-MPs from 87 singleton premature neonates delivered within 48 hours from amniocentesis (gestational age, mean +/- SD: 28.9 +/- 3.3 weeks) were analyzed blindly using strict National Institute of Child Health and Human Development criteria. Strips were evaluated at three time points: at admission, at amniocentesis, and prior to delivery. Intra-amniotic inflammation was established based on a previously validated proteomic fingerprint (mass-restricted score). Diagnoses of histological chorioamnionitis and EONS were based on well-recognized pathological, clinical, and laboratory criteria. We determined that fetuses of women with severe intra-amniotic inflammation had a higher FHR baseline throughout the entire monitoring period and an increased frequency of a nonreactive FHR-MP at admission. Of all FHR-MPs, a nonreassuring test at admission had 32% sensitivity, 95% specificity, 73% positive predictive value, 77% negative predictive value, and 76% accuracy in predicting EONS. Although a nonreassuring FHR-MP at admission was significantly associated with EONS after correcting for gestational age (odds ratio, 5.6; 95% confidence interval, 1.2 to 26.2; P = 0.030), the majority of the neonates that developed EONS had an overall reassuring FHR-MP. Nonreassuring FHR-MPs at either amniocentesis or delivery had no association with EONS. We conclude that in cases complicated by preterm birth, a nonreassuring FHR-MP at the initial evaluation is a specific but not a sensitive predictor of EONS. An abnormal FHR-MP can thus raise the level of awareness that a fetus with EONS may be born, but it is not a useful clinical indicator of the need for antibiotic treatment of the neonate. PMID- 18512203 TI - Anatomic variance in common vascular pedicle of the gracilis and adductor longus muscles: feasibility of double functioning free muscle transplantation with single pedicle anastomosis. AB - Fifty thighs from fresh human cadavers were studied to evaluate the feasibility of a double functioning free muscle transfer of the gracilis and adductor longus with single common vascular pedicle anastomosis. Methylene blue intra-arterial injection and loupe-magnified dissection were used to demonstrate three groups of vascular patterns in these two muscles. The common vascular pedicles of 88% of our specimen muscles were long enough for possible anastomosis. Ten percent (type B2) were quite short, making microsurgical procedure difficult. Two percent (type A3) of our specimens were not suitable for single anastomosis. Four percent of our gracilis muscles had two major arterial pedicles that branched from the common pedicle in a Y-shaped configuration. If only one pedicle of this type is harvested during a free gracilis muscle transfer, it may cause inadequate flap perfusion. Four specimens were studied using contrast media angiography to confirm both are Mathes and Nahai type II muscle flaps. In summary, this study typed the common vascular pedicle of our sample of gracilis and adductor longus muscles and confirmed the feasibility of double functioning free muscle transfer of the gracilis and adductor longus with single vascular anastomosis. PMID- 18512205 TI - Reassembly of anthramycin biosynthetic gene cluster by using recombinogenic cassettes. AB - The reassembly and heterologous expression of complete gene clusters in shuttle vectors has enabled investigations of several large biosynthetic pathways in recent years. With a gene cluster in a mobile construct, the interrogation of gene functions from both culturable and nonculturable organisms is greatly accelerated and large pathway engineering efforts can be executed to produce "new" natural products. However, the genetic manipulation of complete natural product biosynthetic gene clusters is often complicated by their sheer size (10 200 kbp), which makes standard restriction/ligation-based methods impracticable. To circumvent these problems, alternative recombinogenic methods, which depend on engineered homology-based recombination have recently arisen as a powerful alternative. Here, we describe a new general technique that can be used to reconstruct large biosynthetic pathways from overlapping cosmids by retrofitting each cosmid with a "recombinogenic cassette" that contains a shared homologous element and orthogonal antibiotic markers. We employed this technique to reconstruct the anthramycin biosynthetic gene cluster of the thermotolerant actinomycete Streptomyces refuineus, from two >30 kbp cosmids into a single cosmid and integrate it into the genome of Streptomyces lividans. Anthramycin production in the heterologous Streptomyces host confirmed the integrity of the reconstructed pathway and validated the proposed boundaries of the gene cluster. Notably, anthramycin production by recombinant S. lividans was seen only during growth at high temperature--a property also shown by the natural host. This work provides tools to engineer the anthramycin biosynthetic pathway and to explore the connection between anthramycin production and growth at elevated temperatures. PMID- 18512206 TI - Femtosecond transition-state imaging of the A-band CH3I photodissociation. PMID- 18512207 TI - Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes. PMID- 18512208 TI - Life as a nanoscale phenomenon. AB - The nanoscale is not just the middle ground between molecular and macroscopic but a dimension that is specifically geared to the gathering, processing, and transmission of chemical-based information. Herein we consider the living cell as an integrated self-regulating complex chemical system run principally by nanoscale miniaturization, and propose that this specific level of dimensional constraint is critical for the emergence and sustainability of cellular life in its minimal form. We address key aspects of the structure and function of the cell interface and internal metabolic processing that are coextensive with the up scaling of molecular components to globular nanoobjects (integral membrane proteins, enzymes, and receptors, etc) and higher-order architectures such as microtubules, ribosomes, and molecular motors. Future developments in nanoscience could provide the basis for artificial life. PMID- 18512209 TI - Postnatal binge-like alcohol exposure reduces spine density without affecting dendritic morphology in rat mPFC. AB - Among the deficits associated with fetal alcohol syndrome (FAS), cognitive impairments are the most debilitating and permanent. These impairments, including deficits in goal-directed behavior, attention, temporal planning, and other executive functions, could result from damage to the prefrontal cortex (PFC), an area that has not been studied sufficiently in the context of FAS. Neuronal connectivity in this area, as measured by distribution of dendritic spines and the complexity of dendritic tree structure, can be influenced by exogenous variables other than alcohol, and the neuronal connectivity in other brain regions can be affected by alcohol exposure. The goal of this study was to determine whether binge-like alcohol exposure on postnatal days (PD) 4-9 affects dendritic spine density and other dendritic tree parameters in mPFC that could possibly underlie functional damage. Rats were intubated with alcohol [5.25 g/kg/day; alcohol exposed (AE)], sham intubated (SI), or remained with the mother (SC, suckle control) on PD 4-9. Animals were sacrificed between PD 26 and PD 30 and brains were processed for Golgi-Cox staining. Apical dendrite complexity and spine density were evaluated for layer III neurons in the mPFC using NeuroLucida software (MicroBrightField, Inc.). Spine density was significantly decreased in AE animals relative to SI and SC controls, but no differences in dendritic complexity were found across experimental groups. Our findings demonstrate that neonatal alcohol exposure has a persistent effect on the spine density in mPFC that can explain functional deficits in this cortical area. PMID- 18512210 TI - 7-nitroindazole, nitric oxide synthase inhibitor, attenuates physical dependence on butorphanol in rat. AB - Butorphanol is a synthetic opioid agonist/antagonist analgesic agent that mainly exerts its effects through kappa-opioid receptors. It has been demonstrated that kappa-opioid receptors preferentially mediate the development of physical dependence upon butorphanol and the associated withdrawal syndrome. However, it is not fully understood whether or not nNOS-containing neurons in the various brain regions play an important role in butorphanol withdrawal. Therefore, this study was conducted to determine whether the selective nNOS inhibitor, 7-NI, modifies the development of butorphanol withdrawal and changes of nNOS expressions in different brain regions in physically butorphanol-dependent rats. The first part of the study focused on withdrawal behaviors in male Sprague Dawley rats. Physical dependence was induced by a 72-h i.c.v. infusion with butorphanol (26 nmol/mixrol/h) and withdrawal was subsequently precipitated by i.c.v. challenge with naloxone (48 nmol/5 microl/rat) 2 h after termination of the butorphanol infusion. The butorphanol/7-NI coadministration group showed a significant decrease in several signs of withdrawal such as teeth chattering, as compared with the butorphanol-treated group. In the second part of the study, immunohistochemical analysis was performed to determine the expression of nNOS in the various brain regions. In the butorphanol/7-NI coadministration group, the number of cells labeled for nNOS was significantly lower in the various brain regions (including the caudate putamen, nucleus accumbens, and hippocampus) than in the butorphanol group. Therefore, 7-NI decreased in butorphanol-induced physical dependence and nNOS expression. Taken together, these findings suggest that the nNOS system is involved in the development of butorphanol-induced physical dependence, and 7-NI has potential clinical application as a candidate for the treatment of opioid withdrawal syndrome. PMID- 18512211 TI - Surface chemistry of Ag particles: identification of oxide species by aberration corrected TEM and by DFT calculations. PMID- 18512212 TI - Seeing molecules by eye: surface plasmon resonance imaging at visible wavelengths with high spatial resolution and submonolayer sensitivity. PMID- 18512213 TI - Chandelier cartridges in the prefrontal cortex are reduced in isolation reared rats. AB - Chandelier neurons are a subset of parvalbumin containing cortical interneurons characterised by their preferential targeting of the axon initial segments of pyramidal neurons. They have been the focus of recent interest after evidence that the arrays of boutons are reduced in the prefrontal cortex of schizophrenic patients, post mortem. Since one chandelier neuron may innervate the axon initial segments of several hundred pyramidal neurons, it is hypothesized that their special connectivity might facilitate synchronisation of cortical outputs and play a key role in working memory. Disruption in their function is therefore thought to play a potentially important role in cortically associated symptoms of schizophrenia. Using the isolation rearing animal model of schizophrenia, we examined immunolabelling for GABA-transporter 1, a marker of chandelier cartridges. We show that the numbers of arrays of chandelier axons are reduced by 36% in the ventral prelimbic cortex of isolation-reared rats, compared with their socially-housed litter mates. This mimics findings in the PFC of schizophrenic patients where GAT-1-positive cartridges are reduced by 40% and is the first study to demonstrate changes in chandelier cartridges in an animal model of schizophrenia. PMID- 18512214 TI - Serotoninergic dorsal raphe neurons possess functional postsynaptic nicotinic acetylcholine receptors. AB - Very few neurons in the telencephalon have been shown to express functional postsynaptic nicotinic acetylcholine receptors (nAChRs), among them, the noradrenergic and dopaminergic neurons. However, there is no evidence for postsynaptic nAChRs on serotonergic neurons. In this study, we asked if functional nAChRs are present in serotonergic (5-HT) and nonserotonergic (non-5 HT) neurons of the dorsal raphe nucleus (DRN). In rat midbrain slices, field stimulation at the tegmental pedunculopontine (PPT) nucleus evoked postsynaptic currents (eEPSCs) with different components in DRN neurons. After blocking the glutamatergic and GABAergic components, the remaining eEPSCs were blocked by mecamylamine and reduced by either the selective alpha7 nAChR antagonist methyllycaconitine (MLA) or the selective alpha4beta2 nAChR antagonist dihydro beta-eritroidine (DHbetaE). Simultaneous addition of MLA and DHbetaE blocked all eEPSCs. Integrity of the PPT-DRN pathway was assessed by both anterograde biocytin tracing and antidromic stimulation from the DRN. Inward currents evoked by the direct application of acetylcholine (ACh), in the presence of atropine and tetrodotoxin, consisted of two kinetically different currents: one was blocked by MLA and the other by DHbetaE; in both 5-HT and non-5-HT DR neurons. Analysis of spontaneous (sEPSCs) and evoked (eEPSCs) synaptic events led to the conclusion that nAChRs were located at the postsynaptic membrane. The possible implications of these newly described nAChRs in various physiological processes and behavioral events, such as the wake-sleep cycle, are discussed. PMID- 18512215 TI - Dirhodium(II)-catalyzed intramolecular C-H amination of aryl azides. PMID- 18512216 TI - Stereoselective dearomatizing addition of nucleophiles to uncomplexed benzene rings: a route to carbocyclic sugar analogues. PMID- 18512217 TI - Benzene-, pyrrole-, and furan-containing diametrically strapped calix[4]pyrroles- an experimental and theoretical study of hydrogen-bonding effects in chloride anion recognition. PMID- 18512218 TI - Synthesis of marine alkaloids from the oroidin family. PMID- 18512222 TI - Drug delivery trends in clinical trials and translational medicine. PMID- 18512223 TI - Seeking a silver lining: improving the performance of gynecologic cytology in a lower volume setting. PMID- 18512224 TI - Comparison of outcomes for elderly patients treated with weekly paclitaxel in combination with carboplatin versus the standard 3-weekly paclitaxel and carboplatin for advanced nonsmall cell lung cancer. AB - BACKGROUND: The purpose of this study was to compare the outcomes between elderly (aged > or = 70 years) patients treated with paclitaxel on a weekly basis and with carboplatin (every 4 weeks) versus the standard 3-weekly regimen of carboplatin and paclitaxel for first-line therapy of advanced nonsmall cell lung cancer. METHODS: Of the 444 patients enrolled, 136 (31%) were aged > or = 70 years. Seventy-two patients were randomized to the weekly schedule (paclitaxel, 100 mg/m(2) weekly for 3 of 4 weeks; carboplatin, area under the curve [AUC] = 6 mg/mL.min on Day 1 every 4 weeks), and 64 patients were randomized to the standard schedule (paclitaxel, 225 mg/m(2); carboplatin, AUC = 6 mg/mL.min on Day 1 every 21 days). Patients with stable disease or objective response after 4 cycles of therapy were eligible for maintenance therapy with weekly paclitaxel (70 mg/m(2), 3 of 4 weeks). RESULTS: The response rate for elderly patients was 26% on the weekly regimen and 19% on the standard schedule. The median survival duration for the weekly and the standard schedules was 37 weeks and 31 weeks, respectively. The 1-year survival rates were similar at 31% and 33%. Grade 3 to 4 anemia was more common on the weekly schedule (16% vs 6%), whereas grade 3 neuropathy was less common (5.5% vs 9.5%). Nausea and emesis were also less frequent on the weekly schedule. CONCLUSIONS: Efficacy was similar between the weekly regimen and the standard regimen of carboplatin and paclitaxel for elderly patients with advanced NSCLC and may be advantageous based on its favorable tolerability profile. PMID- 18512225 TI - Postnatal ontogeny of the transcription factor Lmx1b in the mouse central nervous system. AB - The expression profile of Lim homeodomain transcription factor Lmx1b in the mouse brain was investigated at different postnatal stages by immunohistochemistry and in situ hybridization. At postnatal day (P) 7, many Lmx1b-expressing neurons were found in the posterior hypothalamic area, supramammillary nucleus, ventral premammillary nucleus, and subthalamic nucleus. In the midbrain, numerous Lmx1b expressing neurons were present in the substantia nigra pars compacta and ventral tegmental area. In the hindbrain, Lmx1b-expressing neurons were primarily observed in the raphe nuclei, parabrachial nuclei, principal sensory trigeminal nucleus, nucleus of the solitary tract, and laminae I-II of the medullary dorsal horn as well as spinal dorsal horn. Although expression levels diminished as postnatal life progressed, persistent expression throughout the first year of life was observed in many of these regions. In contrast, Lmx1b was present in a few brain regions (e.g., principal sensory trigeminal nucleus) only in early life with expression expiring by P60. Lmx1b was observed in dopaminergic neurons in the midbrain and serotonergic neurons in the hindbrain, as determined by double labeling with specific markers. In addition, we found that Lmx1b-expressing neurons are not GABAergic, and Lmx1b was colocalized with Tlx3 in the parabrachial nuclei, principal sensory trigeminal nucleus, nucleus of the solitary tract. as well as the medullary and spinal dorsal horns, suggesting that Lmx1b-expressing cells in these areas are excitatory neurons. Our data suggest that Lmx1b is involved in the postnatal maturation of certain types of neurons and maintenance of their normal functions in the adult brain. PMID- 18512226 TI - A further example of a distinctive autosomal recessive syndrome comprising neonatal diabetes mellitus, intestinal atresias and gall bladder agenesis. AB - We report a patient born to consanguineous parents as a further example of a recently described phenotype comprising neonatal diabetes, intestinal atresias and gall bladder agenesis. Other reports have described cases with overlapping patterns including malrotation, biliary atresia and pancreatic hypoplasia (e.g. as described by Martinez-Frias). We propose that these cases may represent variations of the same syndrome. It is likely that this disorder is inherited as an autosomal recessive trait. Our case is the first to have neonatal diabetes without a demonstrable structural pancreatic abnormality, showing that a deficit in pancreatic function is involved. We sequenced genes with a recognized role in monogenic forms of diabetes, including KCNJ11, ABCC8, GCK, IPF1, HNF1beta, NeuroD1 and TCF7L2, as well as a novel candidate gene, HNF6, known to be involved in hepatobiliary and pancreatic development, but did not identify mutations. PMID- 18512227 TI - Kidney abnormalities in persons with monosomy 15q26. PMID- 18512228 TI - Candidate gene analysis in three families with acilia syndrome. PMID- 18512229 TI - Association of syndromic mental retardation with an Xq12q13.1 duplication encompassing the oligophrenin 1 gene. AB - OPHN1 mutations cause a syndromic form of mental retardation (MR) characterized by cerebellar hypoplasia, early hypotonia, motor and speech delay, with occasional seizures and strabismus. Here we report on a familial chromosome duplication spanning about 800 Kb of Xq12q13.1, associated with MR and a distinctive phenotype in the affected male, but not in his heterozygous mother. The parents were healthy and non-consanguineous with a history of three pregnancies. The first resulted in the birth of a boy with MR, motor impairment and seizures. The second pregnancy was terminated because of trisomy 18. At the time of the third, the first affected boy was analyzed by array-CGH, which revealed a 800 Kb duplication at Xq12q13.1, encompassing three genes, including OPHN1. This mutation was inherited from his healthy mother and was not present in any of the three maternal brothers. To our knowledge this is the first report of a clinical phenotype associated with duplication of Xq12q13. PMID- 18512230 TI - Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an anti-Sox8/9/10 antibody. AB - Quantitative changes of enteric glia (EGC) have been implicated in gastrointestinal disorders. To facilitate future studies of EGC in human pathology, we aimed to characterize thoroughly glial markers in the human enteric nervous system (ENS) and to compare EGC in man and guinea pig. Whole-mount preparations of the enteric nerve plexuses from human and guinea pig ileum and colon were labeled with antibodies against S100b, glial fibrillary acidic protein (GFAP), and p75NGFR and the transcription factors Sox8/9/10 and neuronally counterstained. Abundant immunoreactivity (IR) for S100b, GFAP, p75NGFR, and Sox8/9/10 was detected in EGC of all studied regions. Although the cytoplasmatic staining pattern of most markers did not permit glial quantification, the nuclear localization of Sox8/9/10-IR allowed to identify and count all EGC individually. In both man and guinea pig, myenteric ganglia were larger and contained more EGC and neurons than submucous ganglia. Furthermore, there were more EGC in the human than in the guinea pig myenteric plexus (MP), glial density was consistently higher in the human ENS, and the glia index (glia:neuron ratio) ranged from 1.3 to 1.9 and from 5.9 to 7.0 in the human submucous plexus (SMP) and MP, respectively, whereas, in guinea pig, the glia index was 0.8-1.0 in the SMP and 1.7 in the MP. The glia index was the most robust quantitative descriptor within one species. This is a comprehensive set of quantitative EGC measures in man and guinea pig that provides a basis for pathological assessment of glial proliferation and/or degeneration in the diseased gut. PMID- 18512231 TI - Reply to Hunter's letter on the "misuse of the descriptor "Marfanoid"". PMID- 18512232 TI - Misuse of the descriptor "Marfanoid". PMID- 18512233 TI - Complex toe syndactyly with characteristic facial phenotype: a new syndrome? AB - Nonsyndromic syndactyly is a common, heterogeneous hereditary condition of webbed fingers and toes that can be cutaneous or bony, unilateral or bilateral. We describe a patient with complex toe syndactyly and oligodactyly, some interesting skeletal hand findings and atypical facial features without other case like this described before. Cenani-Lenz syndrome (CLS) is a rare disorder with total syndactyly and irregular synostosis of carpal, metacarpal and phalanges, it may involve ulna and radius and digital rays may be absent, some of these were described with atypical facial features and one patient had renal hypoplasia and vertebral anomalies but our patient does not have the oligodactyly or syndactyly of the hands that is consistently present in all patients with CLS. The atypical facial features of our patient resemble Kabuki syndrome but oligodactyly and complex syndactyly have not been described in Kabuki syndrome and this patient has normal intelligence, and extreme eyelid defect (resembling ablepharon). Therefore, for our patient, we suggested to treat in a new condition of limb anomalies and atypical face. PMID- 18512234 TI - 22q11.2 deletion syndrome in patients admitted to a cardiac pediatric intensive care unit in Brazil. AB - The 22q11.2 deletion syndrome (22q11DS) is one of the most recognizable causes of congenital heart defects (CHDs), but the frequency varies in non-selected populations. The purpose of this study was to determine the incidence and clinical features of patients with CHD and 22q11DS admitted to a pediatric cardiology intensive care unit in Brazil. In a prospective study, we evaluated a consecutive series of 207 patients with a CHD following a clinical protocol and cytogenetic analysis by high resolution karyotype and fluorescent in situ hybridization (FISH). 22q11DS was identified in four patients (2%), a frequency similar to studies that evaluated subjects with major CHDs in other countries. Despite this similarity, we believe that the low rate of prenatal identification of CHDs and the limited access of these patients to appropriate diagnosis and care, which occur in our region, could have had an influence on this frequency. It is possible that 22q11DS patients with a severe CHD could have died before having a chance to access a tertiary hospital, leading to an underestimate of its frequency. PMID- 18512235 TI - Modification of the CHARMM force field for DMPC lipid bilayer. AB - The CHARMM force field for DMPC lipids was modified in order to improve agreement with experiment for a number of important properties of hydrated lipid bilayer. The modification consists in introduction of a scaling factor 0.83 for 1-4 electrostatic interactions (between atoms separated by three covalent bonds), which provides correct transgauche ratio in the alkane tails, and recalculation of the headgroup charges on the basis of HF/6-311(d,p) ab-initio computations. Both rigid TIP3P and flexible SPC water models were used with the new lipid model, showing similar results. The new model in a 75 ns simulation has shown a correct value of the area per lipid at zero surface tension, as well as good agreement with the experiment for the electron density, structure factor, and order parameters, including those in the headgroup part of lipids. PMID- 18512236 TI - Fabrication and characterization of functionally graded nano-micro porous titanium surface by anodizing. AB - The purpose of this study was to fabricate and characterize nanotubular structure on machined titanium (MA) and resorbable blast media (RBM) treated titanium by anodizing. The anodized MA and RBM were characterized with scanning electron microscopy, transmission electron microscopy, X-ray diffraction, energy disperse spectra, X-ray photoelectron spectra, and nano-indentation and scratch test. Highly ordered nanotubular layers of individually 100 nm in diameter and 500 nm in length approximately were formed regardless of the substrates. The nanotubular layers consisted mainly of amorphous TiO(2) with trace fluorine. The nanotubular surfaces on both the substrates significantly reduced water contact angles and elastic modulus compared with those prior to anodizing. The anodizing treatment significantly increased the surface roughness of the smooth MA, but significantly decreased the surface roughness of the roughened RBM. The critical delamination forces of the nanotubular layer were not obtained due to the limitation of surface roughness. The anodized RBM consisted of a nano-micro porous graded structure, or a nanotubular amorphous fluoride containing TiO(2) layer on top of micro-roughened titanium surface, which is expected to significantly improve the surface area that can be used to deliver drugs and growth factors and alleviate the interfacial elastic modulus mismatch as to enhance osseointegration when compared with conventional dental and orthopedic implant devices with smooth or acid etched surface. PMID- 18512237 TI - P53 mutations in stromal fibroblasts sensitize tumors against chemotherapy. AB - The efficacy of chemotherapy is usually viewed as the outcome of cancer-cell autonomous processes while the contribution of stroma is being overseen. Here we show that p53 mutations in stromal fibroblasts, a genetic lesion that is detectable in primary breast, prostate and probably other cancers, while they accelerate tumorigenesis they also sensitize tumours against conventional chemotherapy by doxorubicin and cis-platinum. The mechanism by which p53 of stromal fibroblasts affects the response of a tumour against chemotherapy is likely to involve the induction of senescence in the fibroblasts which in turns results in the production of growth factors acting onto the cancer cells by paracrine mechanisms. Our findings identify stromal fibroblasts as important modulators of the efficacy of anticancer therapy. PMID- 18512238 TI - Vitamin E and cancer: An insight into the anticancer activities of vitamin E isomers and analogs. AB - Current observations in the literature suggest that vitamin E may be a suitable candidate for the adjuvant treatment of cancer. Even though historically most research focused on alpha-tocopherol, more recent evidence suggests that the other isomers of vitamin E (beta-, gamma- and delta-tocopherols and alpha-, beta , gamma- and delta-tocotrienols) differ in their proapoptotic potencies. The main focus of this communication is the current understanding of the molecular mechanisms regulated by vitamin E isomers and their analogs during the induction of apoptosis. This review highlights that the mitochondria are the major target for the induction of apoptosis by vitamin E isomers and analogs and that the various signaling pathways regulated by these agents are likely to contribute towards maximizing the intrinsic pathway of apoptosis triggered initially by the mitochondria. Overall, the presentation of recent studies from the literature in this communication allows the drawing of the following important conclusions: (i) no direct link exists between the antioxidant activity of each isomer/derivative and proapoptotic potency, (ii) tocotrienols are more effective proapoptotic agents than tocopherols, (iii) synthetic modifications of the naturally occurring compounds may improve their apoptotic potency and (iv) vitamin E isomers and derivatives regulate caspase-independent pathways of apoptosis. The latter combined with the evidence presented in this review regarding the additive or synergistic anticarcinogenic effects obtained when vitamin E analogs are used in combination with other cancer chemotherapeutic agents, supports further research to design the most promising vitamin E derivatives and clinically test them in adjuvant chemotherapeutic treatments. PMID- 18512240 TI - The role of PGP9.5 as a tumor suppressor gene in human cancer. AB - PGP9.5 is a controversial molecule from an oncologic point of view. We recently identified frequent methylation of PGP9.5 gene exclusively in primary head and neck squamous cell carcinoma (HNSCC), suggesting that it could be a tumor suppressor gene. On the other hand, PGP9.5 was reported to be overexpressed in a subset of human cancers presumably due to intrinsic oncogenic properties or as a result of transformation. To demonstrate that PGP9.5 possesses tumor suppressive activity, we examined forced expression by stable transfection of PGP9.5 in 4 HNSCC cell lines. Although all 4 cell lines demonstrated reduced log growth rates in culture after transfection, only 2 cell lines with wild type p53 (011, 022) demonstrated decreased growth in soft agar. In 2 cell lines with mutant p53 (013, 019), we observed no altered growth in soft agar and increased sensitivity to UV irradiation. We then tested for and found a high frequency of promoter methylation in a larger panel of primary tumors including HNSCC, esophageal SCC, gastric, lung, prostate and hepatocellular carcinoma. Our data support the notion that PGP9.5 is a tumor suppressor gene that is inactivated by promoter methylation or gene deletion in several types of human cancers. PMID- 18512241 TI - Hypoxia selects for androgen independent LNCaP cells with a more malignant geno- and phenotype. AB - Hypoxia confers resistance to common cancer therapies, however, it has also has been shown to result in genetic alterations which may allow a survival advantage and increase the tumorigenic properties of cancer cells. Additionally, it may exert a selection pressure, allowing expansion of tumor cells with a more aggressive phenotype. To further assess the role of hypoxia in malignant progression in prostate cancer we exposed human androgen dependent prostate cancer cells (LNCaP) to cycles of chronic hypoxia and isolated a subline, LNCaP H1. This article describes the partial characterization of this cell line. The LNCaP-H1 subline showed altered growth characteristics and exhibited androgen independent growth both in vitro and in vivo. Furthermore, these cells were resistant to mitochondrial-mediated apoptosis, probably since the endogenous levels of Bax was lower and Bcl-2 higher than in the parental LNCaP cells. Microarray analysis revealed that a complex array of pathways had differential gene expression between the 2 cell lines, with LNCaP-H1 cells exhibiting a genetic profile which suggests that they may be more likely metastasize to distant organs, especially bone. This was supported by an in vitro invasion assay, and an in vivo metastasis study. This study shows that hypoxia can select for androgen independent prostate cancer cells which have a survival advantage and are more likely to invade and metastasize. PMID- 18512242 TI - Fish consumption and body burden of organochlorines among lower Hudson urban anglers. AB - BACKGROUND: Polychlorinated biphenyls and other organochlorines have contaminated the Hudson River and New York/New Jersey Harbor for many decades. Although body burdens and health effects resulting from exposure to these pollutants have been measured in other populations, little is known about anglers from the lower Hudson area. METHODS: Anglers were recruited from fishing clubs and along piers on the lower Hudson River. One hundred twenty-five participants provided blood samples and questionnaire information. Serum was analyzed at Mount Sinai School of Medicine for organochlorines. RESULTS: Mean concentration of total PCB congeners was 4.78 microg/L. A non-significant adjusted mean difference of -0.59 microg/L was observed between local fish eaters and non-eaters with higher levels among the non-eaters. Similar negative, but non-significant associations were observed for p,p'-DDT, HCB, and beta-BHC. For pp'-DDE, and several chlordane residues, levels were higher among anglers reporting local fish consumption, but the difference was statistically significant only for TN. CONCLUSIONS: These findings suggest eating fish from the lower Hudson River and NY/NJ harbor is not associated with an increased body burden of PCBs or most other organochlorines. PMID- 18512243 TI - A new electromagnetic exposure metric: high frequency voltage transients associated with increased cancer incidence in teachers in a California school. AB - BACKGROUND: In 2003 the teachers at La Quinta, California middle school complained that they had more cancers than would be expected. A consultant for the school district denied that there was a problem. OBJECTIVES: To investigate the cancer incidence in the teachers, and its cause. METHOD: We conducted a retrospective study of cancer incidence in the teachers' cohort in relationship to the school's electrical environment. RESULTS: Sixteen school teachers in a cohort of 137 teachers hired in 1988 through 2005 were diagnosed with 18 cancers. The observed to expected (O/E) risk ratio for all cancers was 2.78 (P = 0.000098), while the O/E risk ratio for malignant melanoma was 9.8 (P = 0.0008). Thyroid cancer had a risk ratio of 13.3 (P = 0.0098), and uterine cancer had a risk ratio of 9.2 (P = 0.019). Sixty Hertz magnetic fields showed no association with cancer incidence. A new exposure metric, high frequency voltage transients, did show a positive correlation to cancer incidence. A cohort cancer incidence analysis of the teacher population showed a positive trend (P = 7.1 x 10(-10)) of increasing cancer risk with increasing cumulative exposure to high frequency voltage transients on the classroom's electrical wiring measured with a Graham/Stetzer (G/S) meter. The attributable risk of cancer associated with this exposure was 64%. A single year of employment at this school increased a teacher's cancer risk by 21%. CONCLUSION: The cancer incidence in the teachers at this school is unusually high and is strongly associated with high frequency voltage transients, which may be a universal carcinogen, similar to ionizing radiation. PMID- 18512244 TI - US estimates of hospitalized pediatric patients with ulcerative colitis: implications for multicenter clinical studies. AB - BACKGROUND: The optimal clinical management of children hospitalized with ulcerative colitis (UC) is evolving. There are limited data quantifying the number of pediatric patients with UC admitted to hospitals in the United States. We analyzed the Kids' Inpatient Database (KID, 2003), to estimate the distribution of hospitalized children with UC and estimate sample sizes available for clinical research. METHODS: We limited our analysis to subjects age less than 18 years. We defined cases of UC as discharge records associated with an ICD-9 code of 556.0-556.9 in the first position. We defined colectomy as principal procedure code of 45.8. We generated weighted estimates for these analyses. To estimate the relationship between number of patients and number of hospitals necessary for clinical trials, we generated 1000 simulated datasets. RESULTS: A total of 2311 UC cases were identified. The mean age at admission was 13.1 (standard error [SE] 0.1) years, and 9% (SE 0.9%) underwent colectomy during their hospitalization. 1008 UC cases were treated at high-volume hospitals; the majority of these children were treated at children's hospitals. Simulation studies suggest that approximately 5 high-volume hospitals would be necessary to generate sample sizes necessary for a pilot clinical trial of refractory UC. CONCLUSIONS: Approximately half of all young patients hospitalized with UC in the US were treated at a limited number of high-volume hospitals, and approximately 5 such centers would be adequate for pilot clinical trials of hospitalized patients with refractory UC. PMID- 18512245 TI - Longitudinal study of quality of life and psychological functioning for active, fluctuating, and inactive disease patterns in inflammatory bowel disease. AB - BACKGROUND: The aim was to assess quality of life (QOL) and psychological functioning in inflammatory bowel disease (IBD) as related to patterns of disease activity over time. METHODS: Study participants were 388 recently diagnosed individuals from the population-based Manitoba IBD Cohort Study. They completed mail-out surveys at 6-month intervals and clinical interviews annually. Based on their 2-year pattern of self-reported disease activity, participants were assigned to 1 of 3 groups: consistently active, fluctuating, or consistently inactive disease. Disease type (Crohn's disease [CD] or ulcerative colitis [UC]) was confirmed through chart review. Change over time was modeled for measures of QOL and positive and negative psychological functioning using mixed-effects regression analyses. RESULTS: Half of the participants had fluctuating disease activity, while almost one-third of participants reported consistent active disease. Participants with the fluctuating activity pattern showed significant improvement in disease-specific QOL compared to participants with consistent activity. Perceived stress, health anxiety, and pain anxiety decreased while pain catastrophizing and mastery increased over time, although the amount of change was not significantly different among disease activity patterns. However, when the data were averaged over time there were significant differences among disease activity patterns on most outcomes. Significant effects of CD versus UC were observed only for the pain measures. CONCLUSIONS: Change in IBD QOL is influenced by one's longitudinal profile of disease activity, but change in psychological functioning is not. Effects of disease activity on psychological functioning were modest, suggesting that disease has an impact even when patients are not experiencing active symptoms. PMID- 18512246 TI - Aberrant innate immune responses in TLR-ligand activated HLA-B27 transgenic rat cells. AB - BACKGROUND: Commensal enteric microbiota initiate and perpetuate immune-mediated colitis in HLA-B27 transgenic (TG) rats but not wildtype (non-TG) littermates. However, the role of the innate immune response to bacterial components has not been established. METHODS: We examined responses induced by bacterial adjuvants through Toll-like receptor (TLR) and NOD2 signaling in T-cell-depleted splenocytes from HLA-B27 TG rats versus non-TG controls. RESULTS: We found that various bacterial adjuvants induced TNF production by cells obtained from specific pathogen-free (SPF) and germ-free (GF, sterile) TG and non-TG rats. Peptidoglycan-polysaccharide (PG-PS), lipopolysaccharide (LPS), and CpG DNA motifs stimulated higher levels of TNF production by SPF TG rat spleen cells compared to non-TG cells. CD11b/c cell depletion eliminated PG-PS and LPS-induced TNF and dramatically reduced CpG-stimulated TNF production. Both SPF and GF TG rat spleens contain more cells that express high levels of CD11b/c and show enhanced mRNA expression of TLR-2 and TLR-4 compared to non-TG rat spleens. In contrast, constitutive and bacterial-induced IL-10 production was markedly lower in TG cells compared to non-TG cells of rats from the same SPF or GF housing conditions. Notably, the ratio of TNF to IL-10 produced after TLR ligand activation was significantly higher in TG than non-TG cells. CONCLUSIONS: HLA-B27 TG rats have an aberrant cell composition, altered functional TLR expression, and an intrinsic defect in IL-10 production in response to TLR ligands, which may result in exaggerated proinflammatory responses to commensal enteric bacteria and uncontrolled inflammation in this colitis model. PMID- 18512247 TI - College adjustment in University of Michigan students with Crohn's and colitis. AB - BACKGROUND: Adjustment to college is critical for academic success. Poor college adjustment correlates with poor academic performance, low graduation rates, and poor success later in life. Limited data are available on the effects of inflammatory bowel disease (IBD) on college adjustment. We hypothesize that disease activity negatively impacts on QOL, and adversely affects college adjustment. METHODS: Undergraduate students (6 Crohn's disease [CD], 12 ulcerative colitis [UC], 19 healthy controls) completed a standardized college adjustment survey (SACQ) and QOL instrument (SF-12). Where appropriate, disease specific activity and QOL indices were obtained (HBI, SCCAI, SIBDQ). RESULTS: There was an inverse correlation between disease activity and college adjustment in CD and UC (R = -0.6554, p = 0.0032). IBD students had lower physical QOL (SF 12) than controls (p = 0.0009). Emotional domain of college adjustment correlated best with SIBDQ (R = 0.8228, p < 0.0001), and correlated better in CD (R = 0.8619) than UC (R = 0.7946). Mental QOL (SF-12) was worse in CD than UC (p = 0.0211), but neither differed from controls (p = 0.4, p = 0.6). CONCLUSIONS: Students with active Crohn's and colitis adjust less well to college life. Physical and emotional factors likely contribute. More aggressive medical therapy and better emotional support before and during college may result in happier and healthier college students, leading to higher graduation rates and future success. Interventions resulting in better disease control and support systems may improve college performance and provide long-term benefits to young adults with IBD. PMID- 18512248 TI - IL-23/IL-17 immunity as a hallmark of Crohn's disease. AB - BACKGROUND: We studied the balance between ileal T-effector cells versus T regulatory cells in active and inactive Crohn's disease (CD). METHODS: We compared effector and regulatory T-cell-related markers such as interleukin (IL) 17, interferon (IFN)-gamma, IL-4, and Foxp3 transforming growth factor (TGF)-beta CTLA-4 and markers for innate immune activation such as IL-6, IL-10, IL-18, IL 23, tumor necrosis factor (TNF)-alpha, and IL-12p70, studied with immunohistochemistry and RT-PCR in ileal biopsies from patients with active or inactive CD and from control subjects. IL-17 in fecal samples was detected by ELISA. The effect of IL-17 on IL-8 and TNF-alpha mRNA expression in epithelial cell line Caco-2 was studied. RESULTS: The numbers of IL-4-, IL-17-, and IL 23(p19)-positive cells in the lamina propria were higher in patients with CD, both active and inactive, than in the controls. mRNA expression of IL-17A, IL-6, and Foxp3 was increased in the biopsies both from patients with active disease and those in remission, whereas mRNA expression of IL-23 was increased only in active disease. Fecal IL-17 concentration was increased in patients with active disease. IL-17 enhanced the IL-8 and TNF-alpha response of the epithelial cell line to lipopolysaccharide (LPS) in vitro. CONCLUSIONS: Our findings suggest that activation of the IL-23/IL-17 axis is fundamentally connected to the etiology of CD and may represent the basis for the relapsing nature of the disease by increasing the sensitivity of epithelium to microbial LPS. PMID- 18512249 TI - Neurons control the expression of connexin 30 and connexin 43 in mouse cortical astrocytes. AB - A characteristic feature of astrocytes is their high level of intercellular communication mediated by gap junctions. The two main connexins, Cx30 and Cx43, that form these junctions in astrocytes of adult brain display different developmental and regional expression, with a delayed onset of appearance for Cx30. In primary cultures of astrocytes from newborn cerebral cortex, while Cx43 is abundantly expressed, Cx30 is not detectable. In the present report, Western blot and confocal immunofluorescence analysis performed in astrocyte/neuron cocultures demonstrate that neurons upregulate the expression of Cx43 and induce that of Cx30 in subsets of astrocytes preferentially located in close proximity to neuronal soma. In Cx43 lacking astrocytes cocultured with neurons, the induction of Cx30 allows the restoration of dye coupling within islets of Cx30 positive astrocytes, indicating that intercellular channels formed by Cx30 are functional. The upregulating effect of neurons on the expression of connexins in cortical astrocytes is independent of their electrical activity and requires tight interactions between both cell types. This effect is reversed after neuronal death induced by neurotoxic treatments. Furthermore, excitotoxic treatments triggering neuronal death in vivo lead to a downregulation of both connexins in reactive astrocytes located within the area depleted in neurons. Altogether these observations indicate that the expression of the two main astrocyte connexins is tightly regulated by neurons. PMID- 18512250 TI - Production, characterization, and efficient transfection of highly pure oligodendrocyte precursor cultures from mouse embryonic neural progenitors. AB - Much current knowledge of oligodendrocyte biology, the myelin-forming cells in the central nervous system, comes from cell culture studies mainly from postnatal rat tissue but mouse cells have been much more difficult to produce in large quantities. We have developed a high yield protocol for production of oligodendrocyte precursor cells from mouse embryonic neural progenitors grown as neurospheres. Neurospheres can be maintained and expanded for long periods in culture in the presence of epidermal growth factor (EGF). When floating neurospheres were plated on substrate-coated dishes in media supplemented with platelet derived growth factor (PDGF) and basic fibroblast growth factor (bFGF), the spheres attached and generated migrating cells that were predominantly oligodendrocyte-lineage cells. Furthermore, cells in spheres could be shifted to the oligodendrocyte phenotype prior to plating on substrate, by incubation in suspension with PDGF/bFGF. Single cell suspensions plated after dissociation of either EGF-treated neurospheres or PDGF/bFGF-treated oligospheres had the bipolar, elongated morphology characteristic of oligodendrocyte precursor cells. mRNA and protein expression analysis of the cells generated by this method confirmed their oligodendrocyte lineage. Oligodendrocyte precursors generated by this method matured in response to ciliary neurotrophic factor treatment, producing cells with multiple processes and myelin-like membranes. The most important aspect of this protocol is the ability to generate very high numbers of relatively pure mouse oligodendrocyte progenitor cells, which can be easily transfected. These studies open up many kinds of investigations on transgenic and mutant mouse oligodendrocytes, thereby providing a valuable tool to study oligodendrocyte biology and development. PMID- 18512251 TI - Beta-amyloid peptide stimulates endozepine release in cultured rat astrocytes through activation of N-formyl peptide receptors. AB - Astroglial cells synthesize and release endozepines, a family of neuropeptides derived from diazepam-binding inhibitor (DBI). The authors have recently shown that beta-amyloid peptide (Abeta) stimulates DBI gene expression and endozepine release. The purpose of this study was to determine the mechanism of action of Abeta in cultured rat astrocytes. Abeta(25-35) and the N-formyl peptide receptor (FPR) agonist N-formyl-Met-Leu-Phe (fMLF) increased the secretion of endozepines in a dose-dependent manner with EC(50) value of approximately 2 microM. The stimulatory effects of Abeta(25-35) and the FPR agonists fMLF and N-formyl-Met Met-Met (fMMM) on endozepine release were abrogated by the FPR antagonist N-t-Boc Phe-Leu-Phe-Leu-Phe. In contrast, Abeta(25-35) increased DBI mRNA expression through a FPR-independent mechanism. Abeta(25-35) induced a transient stimulation of cAMP formation and a sustained activation of polyphosphoinositide turnover. The stimulatory effect of Abeta(25-35) on endozepine release was blocked by the adenylyl cyclase inhibitor somatostatin, the protein kinase A (PKA) inhibitor H89, the phospholipase C inhibitor U73122, the protein kinase C (PKC) inhibitor chelerythrine and the ATP binding cassette transporter blocker glyburide. Taken together, these data demonstrate for the first time that Abeta(25-35) stimulates endozepine release from rat astrocytes through a FPR receptor positively coupled to PKA and PKC. PMID- 18512252 TI - Reduced synaptic activity precedes synaptic stripping in vagal motoneurons after axotomy. AB - Activated microglia, which spread on the motor neurons following nerve injury, engage in the displacement of detached afferent synaptic boutons from the surface of regenerating motor neurons. This phenomenon is known as "synaptic stripping." The present study attempted to examine whether changes in the synaptic inputs after motor nerve injury correlated with the microglial attachment to the dorsal motor neurons of the vagus (DMV). DMV neurons in Wistar rats could survive after nerve injury, whereas most of injured DMV neurons in the C57BL/6 mice died. At 2 days after nerve injury, a significant decrease was observed in the frequencies of both spontaneous and miniature EPSCs and IPSCs recorded from DMV neurons in the slice preparation but not from the mechanically dissociated neurons in the Wistar rats. At this stage, no direct apposition of microglia on the injured neurons was observed. High-K(+) stimulation restored their frequencies to control levels. Furthermore, PPADS and DPCPX, antagonists of P2 and adenosine receptors, respectively, also stimulated the recovery of their frequencies. In contrast, no significant change was detected in the spontaneous EPSCs frequency recorded from the severely injured DMV neurons in the slice preparation of the C57BL/6 mice. These observations strongly suggest that presynaptic inhibition through glia derived ATP and adenosine, thus precedes synaptic stripping in regenerating DMV neurons following nerve injury. PMID- 18512253 TI - A versatile chiral selector for determination of enantiomeric composition of fluorescent and nonfluorescent chiral molecules using steady-state fluorescence spectroscopy. AB - A fluorescent chiral molecular micelle (FCMM), poly (sodium N-undecanoyl-L phenylalaninate) (poly-L-SUF), was developed as a chiral selector for enantiomeric recognition and determination of enantiomeric composition of four fluorescent and four nonfluorescent chiral molecules by use of steady-state fluorescence spectroscopy. The influence of FCMM concentration, buffer pH and complexation medium on FCMM-analyte host-guest complexation, and the emission spectral properties of the resulting complexes were investigated. The chiral interactions of the analytes,1,1'-binaphthyl-2,2'-diamine, 1-(9-anthryl)-2,2,2 trifluoroethanol, propranolol, naproxen, chloromethyl menthyl ether (CME), citramalic acid, tartaric acid, and limonene (LIM), in the presence of poly-L-SUF were based on diastereomeric complex formation. The figures of merit obtained from the partial-least-squares regression modeling of the calibration samples suggested good prediction ability for the validation of six of the eight chiral analytes. Better host-guest complexation of the more hydrophobic molecules, CME and LIM, were obtained in methanol/water mixtures, resulting in better predictability of the regression models. Prediction ability of the models was evaluated by use of the root-mean-square percent relative error (RMS%RE) and was found to range from 1.77 to 15.80% (buffer), 1.26 to 7.95% (25:75 methanol/water), and 1.21 to 4.28% (75:25 methanol/water). PMID- 18512254 TI - Single-molecule chiroptical spectroscopy: Fluorescence excitation of individual helicene molecules in polymer-supported thin-films. AB - We present results of fluorescence excitation circular dichroism studies of the chiroptical response of single (bridged triarylamine) helicene molecules immobilized at a polymer interface. We extract directly dissymmetry parameters, and corresponding probability distributions, associated with the single-molecule fluorescence excitation associated with modulation of a circular polarized excitation field for three different excitation wavelengths (405, 440, 457 nm) showing circular dichroism in bulk films. The observed single molecule chiroptical response is anomalously large in comparison with the results of time dependent density functional calculations, and the observed defocused emission patterns seem to indicate a higher multipole nature to the transition probed. Our results provide new insights into chiroptical properties of chiral fluorophores that are hidden under the extensive averaging associated with conventional chiroptical probes. PMID- 18512255 TI - Antioxidant protection during ageing and senescence in chloroplasts of tobacco with modulated life span. AB - We studied changes in antioxidant protection during ageing and senescence in chloroplasts of tobacco (Nicotiana tabacum L., cv. Wisconsin) with introduced SAG(12) promoter fused with ipt gene for cytokinin synthesis (transgenic plants with increased levels of cytokinins, SAG) or without it (control). Old leaves of SAG plants as well as their chloroplasts maintained higher physiological parameters compared to controls; accordingly, we concluded that their ageing was diverted due to increased cytokinin content. The chloroplast antioxidant protection did not decrease as well. Although antioxidant protection usually decreased in whole leaves of senescing control plants, ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) activity, which maintained the high redox state of ascorbate, increased in chloroplasts of old control leaves. PMID- 18512256 TI - Use of alternative therapies in patients with dementia and mild cognitive impairment: a prospective, controlled study. AB - OBJECTIVES: To study the frequency and type of alternative therapies used in patients with dementia and Mild Cognitive Impairment. METHODS: In a prospective study, 77 outpatients and 62 healthy volunteers were interviewed using a questionnaire concerning frequency, type and motivation of the use of alternative therapies. RESULTS: 47% of patients and 18% of control subjects used alternative therapies to treat or prevent disturbances of memory (p < 0.001). No correlation could be found between use of alternative medicine and professional education, severity of disease, religion or health insurance status (p > 0.05). The most commonly used substances were vitamins. Side effects remained unreported. 52% of patients were treated with conventional medications. Only 44% of the patients informed their physician about the use of alternative medicine. CONCLUSIONS: The use of alternative therapies, particularly vitamins, and herbal preparations of unknown efficacy is common among patients with dementia and MCI. Hence, the treating physician should actively inquire about their use as some preparations have pharmacological activity or could interact with other prescribed medications. PMID- 18512257 TI - Neuropsychiatric symptoms and quality of life in patients in the final phase of dementia. AB - OBJECTIVES: To assess neuropsychiatric symptoms and quality of life in a group of patients in the final phase of dementia. METHODS: All patients with dementia (n = 216) residing on dementia special care units of two Dutch nursing homes were included in the study provided they met the criteria for the final phase of dementia. Neuropsychiatric symptoms were assessed with the Neuropsychiatric Inventory Nursing Home version (NPI-NH) and the Cohen Mansfield Agitation Inventory (CMAI). Quality of life was assessed with the QUALIDEM. RESULTS: Of the 216 dementia patients 39 met the criteria for the final phase of dementia. The patients showed a specific pattern of behaviours with a high prevalence of apathy, agitation and behaviours that were mainly observed during morning care such as making strange noises, grabbing, performing repetitious mannerism, spitting, hitting, screaming and pushing. Overall quality of life of these patients in the final phase of dementia was moderate. CONCLUSION: In this small sample, patients in the final phase of dementia show specific behavioural problems, that mainly should be addressed with psychosocial interventions. PMID- 18512258 TI - Controlling kinesin motor proteins in nanoengineered systems through a metal binding on/off switch. AB - A significant challenge in utilizing kinesin biomolecular motors in integrated nanoscale systems is the ability to regulate motor function in vitro. Here we report a versatile mechanism for reversibly controlling the function of kinesin biomolecular motors independent of the fuel supply (ATP). Our approach relied on inhibiting conformational changes in the neck-linker region of kinesin, a process necessary for microtubule transport. We introduced a chemical switch into the neck-linker of kinesin by genetically engineering three histidine residues to create a Zn(2+)-binding site. Gliding motility of microtubules by the mutant kinesin was successfully inhibited by >/=10 microM Zn(2+), as well as other divalent metals. Motility was successfully restored by removal of Zn(2+) using a number of different chelators. Lastly, we demonstrated the robust and cyclic nature of the switch using sequential Zn(2+)/chelator additions. Overall, this approach to controlling motor function is highly advantageous as it enables control of individual classes of biomolecular motors while maintaining a consistent level of fuel for all motors in a given system or device. PMID- 18512259 TI - Cytocompatibility, interactions, and uptake of polyethyleneimine-coated boron nitride nanotubes by living cells: confirmation of their potential for biomedical applications. AB - Boron nitride nanotubes (BNNTs) have unique physical properties, which can be exploited in the biomedical field. Hence, the surprising lack of reported studies on their biocompatibility and interactions with living cells, addressed by the present paper which deals the results of such an investigation based on 72 h culture of human neuroblastoma cell line (SH-SY5Y) in the presence of an aqueous suspension of polyethyleneimine (PEI)-coated BNNTs. BNNTs conjugated with fluorescent markers (quantum dots) are employed to enable tracking of their uptake by living cells. The results demonstrate good cytocompatibility together with unequivocal BNNT cellular uptake by an energy-dependent endocytic process. PMID- 18512260 TI - Comparison of microbial community composition and activity in sulfate-reducing batch systems remediating mine drainage. AB - Five microbial inocula were evaluated in batch tests for the ability to remediate mine drainage (MD). Dairy manure (DM), anaerobic digester sludge, substrate from the Luttrell (LUTR) and Peerless Jenny King (PJK) sulfate-reducing permeable reactive zones (SR-PRZs) and material from an MD-treatment column that had been inoculated with material from a previous MD-treatment column were compared in terms of sulfate and metal removal and pH neutralization. The microbial communities were characterized at 0, 2, 4, 9, and 14 weeks using denaturing gradient gel electrophoresis and quantitative polymerase chain reaction to quantify all bacteria and the sulfate-reducing bacteria of the genus Desulfovibrio. The cultures inoculated with the LUTR, PJK, and DM materials demonstrated significantly higher rates of sulfate and metal removal, and contained all the microorganisms associated with the desired functions of SR-PRZs (i.e., polysaccharide degradation, fermentation, and sulfate reduction) as well as a relatively high proportion of Desulfovibrio spp. These results demonstrate that inoculum influences performance and also provide insights into key aspects of inoculum composition that impact performance. This is the first systematic biomolecular examination of the relationship between microbial community composition and MD remediation capabilities. PMID- 18512261 TI - Implications of decoupling the intracellular and extracellular levels in multi level models of virus growth. AB - Virus infections are characterized by two distinct levels of detail: the intracellular level describing how viruses hijack the host machinery to replicate, and the extracellular level describing how populations of virus and host cells interact. Deterministic, population balance models for viral infections permit incorporation of both the intracellular and extracellular levels of information. In this work, we identify assumptions that lead to exact, selective decoupling of the interaction between the intracellular and extracellular levels, effectively permitting solution of first the intracellular level, and subsequently the extracellular level. This decoupling leads to (1) intracellular and extracellular models of viral infections that have been previously reported and (2) a significant reduction in the computational expense required to solve the model. However, the decoupling restricts the behaviors that can be modeled. Simulation of a previously reported multi-level model demonstrates this decomposition when the intracellular level of description consists of numerous reaction events. Additionally, examples demonstrate that viruses can persist even when the intracellular level of description cannot sustain a steady-state production of virus (i.e., has only a trivial equilibrium). We expect the combination of this modeling framework with experimental data to result in a quantitative, systems-level understanding of viral infections and cellular antiviral strategies that will facilitate controlling both these infections and antiviral strategies. PMID- 18512262 TI - Practical identifiability of biokinetic parameters of a model describing two-step nitrification in biofilms. AB - Parameter estimation and model calibration are key problems in the application of biofilm models in engineering practice, where a large number of model parameters need to be determined usually based on experimental data with only limited information content. In this article, identifiability of biokinetic parameters of a biofilm model describing two-step nitrification was evaluated based solely on bulk phase measurements of ammonium, nitrite, and nitrate. In addition to evaluating the impact of experimental conditions and available measurements, the influence of mass transport limitation within the biofilm and the initial parameter values on identifiability of biokinetic parameters was evaluated. Selection of parameters for identifiability analysis was based on global mean sensitivities while parameter identifiability was analyzed using local sensitivity functions. At most, four of the six most sensitive biokinetic parameters were identifiable from results of batch experiments at bulk phase dissolved oxygen concentrations of 0.8 or 5 mg O(2)/L. High linear dependences between the parameters of the subsets (KO2,AOB,muAOB) and (KO2,NOB,muNOB) resulted in reduced identifiability. Mass transport limitation within the biofilm did not influence the number of identifiable parameters but, in fact, decreased collinearity between parameters, especially for parameters that are otherwise correlated (e.g., muAOB) and KO2,AOB, or muNOB and KO2,NOB). The choice of the initial parameter values had a significant impact on the identifiability of two parameter subsets, both including the parameters muAOB and KO2,AOB. Parameter subsets that did not include the subsets muAOB and KO2,AOB or muNOB and KO2,NOB were clearly identifiable independently of the choice of the initial parameter values. PMID- 18512263 TI - Cloning, expression, and characterization of novel thermostable family 7 cellobiohydrolases. AB - As part of the effort to find better cellulases for bioethanol production processes, we were looking for novel GH-7 family cellobiohydrolases, which would be particularly active on insoluble polymeric substrates and participate in the rate-limiting step in the hydrolysis of cellulose. The enzymatic properties were studied and are reported here for family 7 cellobiohydrolases from the thermophilic fungi Acremonium thermophilum, Thermoascus aurantiacus, and Chaetomium thermophilum. The Trichoderma reesei Cel7A enzyme was used as a reference in the experiments. As the native T. aurantiacus Cel7A has no carbohydrate-binding module (CBM), recombinant proteins having the CBM from either the C. thermophilum Cel7A or the T. reesei Cel7A were also constructed. All these novel acidic cellobiohydrolases were more thermostable (by 4-10 degrees C) and more active (two- to fourfold) in hydrolysis of microcrystalline cellulose (Avicel) at 45 degrees C than T. reesei Cel7A. The C. thermophilum Cel7A showed the highest specific activity and temperature optimum when measured on soluble substrates. The most effective enzyme for Avicel hydrolysis at 70 degrees C, however, was the 2-module version of the T. aurantiacus Cel7A, which was also relatively weakly inhibited by cellobiose. These results are discussed from the structural point of view based on the three-dimensional homology models of these enzymes. PMID- 18512264 TI - Recent changes in drug poisoning mortality in the United States by urban-rural status and by drug type. AB - PURPOSE: This study was conducted to determine how the recently reported increase in drug poisoning mortality rates in the United States varied by degree of urbanization. Although drug poisoning is traditionally seen as an urban problem, evidence suggested that at least one component of the recent increase, deaths involving opioid analgesics, was increasing more rapidly in rural areas. METHODS: The study compared age-adjusted unintentional and undetermined drug poisoning mortality rates between 1999 and 2004 from the National Vital Statistics System (NVSS) in each of six urban-rural categories. RESULTS: Unintentional and undetermined drug poisoning mortality rates rose 62% from 1999 to 2004. Metropolitan county rates rose 51%, an increase of 2.66/100,000, while nonmetropolitan county rates rose 159%, an increase of 4.81/100,000. By 2004, metropolitan and nonmetropolitan drug poisoning rates had roughly equalized. In the narcotic drug category, which included heroin, cocaine, and opioid analgesics, the most urban ("large central metro") counties increased only 16% while the most rural ("noncore, nonmetropolitan") counties increased 248%. Heroin rates did not increase significantly for any urban-rural category. Cocaine rate increases were largest in nonmetropolitan counties. Opioid analgesic rate increases ranged from a low of 52% in large central metro counties to an increase of 371% in nonmetropolitan, noncore counties. CONCLUSIONS: Prescription drugs have replaced heroin and cocaine as the leading drugs involved in fatal drug overdoses in all urban-rural categories. Fatal drug overdoses are no longer a predominantly urban phenomenon. National prevention efforts will have to shift to address nontraditional populations using nontraditional drugs. PMID- 18512265 TI - The influence of the RGD peptide motif and its contextual presentation in PEG gels on human mesenchymal stem cell viability. AB - An investigation was undertaken into the method of delivery of RGD peptide motifs to adult human mesenchymal stem cells (hMSCs) encapsulated in poly(ethylene glycol) (PEG) hydrogel systems. Previous studies have shown that the viability of hMSCs encapsulated in bio-inert hydrogels, such as PEG-based gels, decreases over time. hMSCs are an adhesive-dependent cell type, requiring attachment sites to maintain their survival and function. The incorporation of tethered RGD peptide motifs was found to sustain a high level of hMSC survival in PEG gels. However, previous reports are largely limited to pendantly tethered RGD in gels; therefore, further investigation into hMSCs' affinity for and response to the contextual presentation of RGD was studied using varying methods of RGD attachment to the PEG gel, as well as delivery of soluble RGD peptides. Studies with encapsulated hMSCs showed that the constrained RGD peptide, bound via two links to the PEG gel, promoted approximately 60% cell survival, while tethering the RGD as a pendant group, bound via a single link to the PEG gel, promoted approximately 80% cell survival. Interestingly, incorporating a glycine spacer arm to the RGD pendant tether further enhanced survival to approximately 88%. Investigations with solubly delivered peptides resulted in a dramatic decrease in cell viability with time, eventually leading to survival that was similar to that of unmodified PEG gels. Integrin staining for alphavbeta3 and alpha4, as well as focal adhesion staining, correlates to the trends in hMSC survival for covalently bound RGD motifs and a loss of viability for the solubly delivered peptide systems. PMID- 18512266 TI - Polyethyleneimine-mediated gene delivery into rat pheochromocytoma PC-12 cells. AB - In this study, we examined the use of polyethyleneimine (PEI) as a non-viral gene carrier and lipofectamine(trade mark) 2000 as control for rat pheochromocytoma PC 12 cells. The complex formation of PEI and DNA or lipofectamine and DNA was characterized by gel electrophoresis and measurement of particle size and surface charge. A gradual increase in surface charge (from 0.7 to 43 mV) and a gradual decrease in particle size (from 900 to 130 nm) was observed in the PEI-DNA complex with higher PEI concentrations. The cytotoxicity of PC-12 cells for lipofectamine-DNA complex was similar to PEI-DNA complex at N:P charge ratios of 4 and 8. Transfection efficiency was 14% for lipofectamine and 15% for PEI. At low N:P ratio, DNA condenses poorly, so the particle size tends to be large and polydispersed, resulting in poor transfection efficiency. Meanwhile, a high N:P ratio results in high transfection efficiency and cytotoxicity. Transfected PC-12 cells showed the generation of neurites from transfected PC-12 cells in the presence of NGF, indicating the differentiation of PC-12 cells. NGF differentiated PC-12 cells were transfected by PEI-DNA complex of N:P charge ratio 8. From real-time imaging for transfection, the enhanced green fluorescent protein (EGFP) started to localize in the nuclei of PC-12 cells at 5 h and localized in the cytoplasm from 15 h. Our study demonstrates that PEI or lipofectamine may be applied as an effective gene carrier for PC-12 cells. PMID- 18512267 TI - Effects of electrical stimulation in C2C12 muscle constructs. AB - Electrical stimulation affects the deposition of extracellular matrices and cellular differentiation. Type I collagen is one of the most abundant extracellular matrix proteins; however, not much is known about the effects of electrical stimulation on collagen type I deposition in C2C12 cells. Thus, we studied the effects of electrical voltage and stimulation frequency in 3D cultured C2C12 muscle cells in terms of metabolic activity, type I collagen deposition and cell morphology. Electrically excitable C2C12 muscle cells were seeded in collagen scaffolds and stimulated with rectangular signals of voltage (2, 5, 7 V) and frequency (1, 2 Hz), using parallel carbon electrodes spaced 1 cm apart. Metabolic activity was quantified by the glucose:lactate concentration ratio in the medium. Apoptotic activity was assessed by TUNEL staining and changes in collagen deposition were identified by immunohistology. The ultrastructure of the tissue was examined by TEM. Glucose and lactate analysis indicated that all groups had similar metabolic activity. TUNEL stain showed no significant difference in apoptotic damage induced by electrical stimulation compared to the control. Samples stimulated at 2 Hz exhibited reduced collagen deposition compared to the control and 1 Hz stimulated samples. Muscle-protein marker desmin was highly expressed in constructs stimulated with 1 Hz/5 V sample. TEM revealed that the stimulated samples developed highly organized sarcomeres, which coincided with improved contractile properties in the 1 Hz/5 V- and 2 Hz/5 V-stimulated groups. Our data implicate that a specific electrical frequency may modulate type I collagen accumulation and a specific voltage may affect the differentiation of muscle sarcomeres in excitable cells. PMID- 18512268 TI - Desferrioxamine-driven upregulation of angiogenic factor expression by human bone marrow stromal cells. AB - Bone marrow stromal cells (BMSCs) are the subject of intense research because of their biological properties and potential use for the repair of damaged tissues. Success of BMSC-based therapies, however, relies on a number of methodological improvements, including the establishment of a vascular network providing nutrients and oxygen to the transplanted cells and ensuring their immediate survival and long-term functionality. We described a method to enhance the autocrine expression of angiogenic factors by BMSCs. For this purpose, human BMSCs were treated with desferrioxamine (DFX). No PDGF-BB, VEGF-R1 or -R2 mRNA expression was detected under any of the conditions tested. mRNA and protein expression levels of TGFbeta1 were similar in BMSCs, whether they were exposed to DFX (50 microM) or to control conditions under normoxia for 48 h. In comparison with the results obtained with control conditions under normoxia, exposure of BMSCs to DFX for 48 h resulted in upregulation of bFGF at the protein (26-fold) but not at the mRNA levels and VEGF at both the mRNA (1.5-fold) and protein levels (4.5-fold). In comparison with the results obtained with control conditions under hypoxia, DFX induced a 50% increase in VEGF secretion but led to the same level of hypoxia inducible factor-1alpha protein expression (a transduction factor involved in angiogenic factor expression and known to be activated by DFX). Exposure of BMSCs to DFX resulted in oversecretion of angiogenic factors, suggesting that DFX-treated BMSCs could be used to supply angiogenic factors. PMID- 18512269 TI - A biomimetic scaffold for culturing limbal stem cells: a promising alternative for clinical transplantation. AB - Limbal tissues can be cultured on various types of scaffolds to create a sheet of limbal-corneal epithelium for research as well as clinical transplantation. An optically clear, biocompatible, biomimetic scaffold would be an ideal replacement graft for transplanting limbal stem cells. In this study, we evaluated the physical and culture characteristics of the recombinant human cross-linked collagen scaffold (RHC-III scaffold) and compared it with denuded human amniotic membrane (HAM). Optical/mechanical properties and microbial susceptibility were measured for the scaffolds. With the approval of the institutional review board, 2 mm fresh human limbal tissues were cultured on 2.5 x 2.5 cm(2) scaffolds in a medium containing autologous serum in a feeder cell-free submerged system. The cultured cell systems were characterized by morphology and immunohistochemistry for putative stem cells and differentiated cell markers. The refractive index (RI) and tensile strength of the RHC-III scaffold were comparable to human cornea, with delayed in vitro degradation compared to HAM. RHC-III scaffolds were 10-fold less susceptible to microbial growth. Cultures were initiated on day 1, expanded to form a monolayer by day 3 and covered the entire growth surface in 10 days. Stratified epithelium on the scaffolds was visualized by transmission electron microscopy. The cultured cells showed p63 and ABCG2 positivity in the basal layer and were immunoreactive for cytokeratin K3 and K12 in the suprabasal layers. RHC-III scaffold supports and retains the growth and stemness of limbal stem cells, in addition to resembling human cornea; thus, it could be a good replacement scaffold for growing cells for clinical transplantation. PMID- 18512270 TI - Resolved: being fat is good for dialysis patients: the Godzilla effect: con. PMID- 18512271 TI - From rising star to nurse leader. PMID- 18512272 TI - Open label trial of alefacept in palmoplantar pustular psoriasis. AB - BACKGROUND: Palmoplantar pustular psoriasis (PPP) is difficult to treat. We assessed the effectiveness of alefacept in PPP and the safety of a 30 mg/week dose. METHODS: Fifteen individuals with PPP were started on 15 mg/week intramuscularly (IM) alefacept. Efficacy was measured by the PPP severity instrument (PSI). Treatment was continued for 16 weeks, and the alefacept dose was increased to 30 mg/week IM at week 9 if the PSI did not decrease by at least 25%. Other outcomes included physician's global assessment (PGA), reported adverse events and CD4+ T-lymphocyte counts. Clinical response was observed for 12 weeks after the last injection. RESULTS: The severity of PPP improved in both the PSI and the PGA (p<0.0001 and p = 0.0009, respectively). Much of the improvement occurred after 10 weeks of therapy. Nail severity scores improved (p = 0.0003). CD4+ counts decreased, but all remained >250 cells/mm3. There were no severe adverse effects or discontinuations due to adverse events. CONCLUSIONS: Alefacept in doses up to 30 mg/week was well tolerated in patients with PPP and appeared to have some efficacy. The use of concomitant therapy, the lack of a comparator, and the small sample size are limitations of the study. PMID- 18512273 TI - Local paclitaxel delivery in peripheral vascular disease. PMID- 18512274 TI - Cardiovascular events during World Cup soccer. PMID- 18512275 TI - Cardiovascular events during World Cup soccer. PMID- 18512276 TI - Cardiovascular events during World Cup soccer. PMID- 18512277 TI - Hutchinson-Gilford progeria syndrome. PMID- 18512278 TI - Effect of cost sharing on screening mammography. PMID- 18512279 TI - Systemic lupus erythematosus. PMID- 18512280 TI - [Tuberculosis meningitis in a Senegalese pediatric hospital: report of 14 cases]. PMID- 18512281 TI - Evolutionists counter creationist's views. PMID- 18512282 TI - Evolutionists counter creationist's views. PMID- 18512283 TI - Evolutionists counter creationist's views. PMID- 18512284 TI - Proteomics-based strategies in kinase drug discovery. AB - Studies of drug action classically assess biochemical activity in settings which typically contain the isolated target only. Recent technical advances in mass spectrometry-based analysis of proteins have enabled the quantitative analysis of sub-proteomes and entire proteomes, thus initiating a departure from the traditional single gene--single protein--single target paradigm. Here, we review chemical proteomics-based experimental strategies in kinase drug discovery to analyse quantitatively the interaction of small molecule compounds or drugs with a defined sub-proteome containing hundreds of protein kinases and related proteins. One novel approach is based on 'Kinobeads'--an affinity resin comprised of a cocktail of immobilized broad spectrum kinase inhibitors--to monitor quantitatively the differential binding of kinases and related nucleotide-binding proteins in the presence and absence of varying concentrations of a lead compound or drug of interest. Differential binding is detected by high throughput and sensitive mass spectroscopy techniques utilizing isobaric tagging reagents (iTRAQ), yielding quantitative and detailed target binding profiles. The method can be applied to the screening of compound libraries and to selectivity profiling of lead compounds directly against their endogenously expressed targets in a range of cell types and tissue lysates. In addition, the method can be used to map drug-induced changes in the phosphorylation state of the captured sub proteome, enabling the analysis of signalling pathways downstream of target kinases. PMID- 18512285 TI - Molecular mechanisms that control leukocyte extravasation through endothelial cell contacts. AB - Leukocyte extravasation and entry into tissue forms the basis for inflammatory reactions and lymphocyte surveillance. After docking at the blood vessel wall at sites of exit leukocytes migrate through the endothelial cell layer and the underlying basement membrane, a process described as diapedesis. In recent years, several endothelial membrane proteins that which participate in this process have been identified. This review focuses on three membrane proteins located at endothelial cell contacts that are involved in the regulation of leukocyte diapedesis. The endothelial cell selective adhesion molecule (ESAM) at endothelial tight junctions and the vascular endothelial receptor-type protein tyrosine phosphatase (VE-PTP), a protein associating with VE-cadherin, both seem to control the integrity of endothelial cell contacts during diapedesis. CD99 and the distantly related CD99L2 are leukocyte membrane proteins that do not belong to any known protein family. They are expressed at endothelial cell contacts and participate in the migration of leukocytes through endothelium and basement membrane. PMID- 18512287 TI - The intrauterine environment, temperament, and development: including the biological foundations of individual differences in the study of psychopathology and wellness. PMID- 18512288 TI - The challenge of mental health research in juvenile justice. PMID- 18512290 TI - Applied brain imaging. PMID- 18512291 TI - Epigenetics: behavioral influences on gene function, part I. Maternal behavior permanently affects adult behavior in offspring. PMID- 18512292 TI - What is an image? PMID- 18512293 TI - Money talks: becoming more comfortable with understanding a family's finances. PMID- 18512294 TI - High waist: hip ratio a marker for atherosclerosis. PMID- 18512295 TI - Increased stress for civil servants and GPs? PMID- 18512296 TI - [Cochlear implantation with suprameatal approach in Chinese children]. AB - OBJECTIVE: To investigate the technique of the suprameatal approach for cochlear implantation in Chinese profound sensory hearing loss children. METHODS: Suprameatal approach for cochlear implantation were used in 50 cases (total 53 ears) with profound sensory hearing loss from May 2005 to January 2007. The electrode was passed through the suprameatal tunnel and went between the incus and chorda tympani into the scala tympani. RESULTS: Electrodes were completely inserted in 51 ears. There were no postoperative complications in all cases. Although the long effect need to be observed, all cases received better hearing and speech development benefit from cochlear implantation in the follow-up period. Among the 50 cases, 26 had speech perception in the open condition; 18 patients could speak short sentences although not clearly; and 6 patients learned to speak individual words only. CONCLUSIONS: The suprameatal approach was found to be a simple and safe technique that does not need mastoidectomy and avoid endangering the facial nerve and the chorda tympani. It enables wide exposure of middle ear and is especially suitable for cases with narrow facial recess or anteriorly located facial nerve. PMID- 18512297 TI - Awareness key in managing depression in later life. PMID- 18512298 TI - Biosecurity, public health and animal welfare. PMID- 18512299 TI - [Picture of the month: giant cell arteritis]. PMID- 18512300 TI - [Whiplash injuries and vulnerability]. PMID- 18512301 TI - [The role of the police in involuntary commitment]. PMID- 18512302 TI - Helping new nurses be assertive about patient safety. PMID- 18512303 TI - Surgical management of diabetic foot infections and amputations. AB - The incidence of diabetes with severe foot infections (eg, necrotizing fasciitis, gas gangrene, ascending cellulitis, infection with systemic toxicity or metabolic instability) has risen significantly during the past decade. Foot infections are a major cause of hospitalization and subsequent lower extremity amputation among patients with diabetes mellitus who have a history of a preexisting ulceration. Surgical management often is required to address severe diabetic foot infections because they can be limb- or life-threatening. Critical limb ischemia, neuropathy, and an immunocompromised host, which often are associated with diabetic foot infections, complicate treatment and are associated with a poorer prognosis. PMID- 18512304 TI - Plastic surgery reconstruction of the diabetic foot. AB - Soft tissue reconstruction of the diabetic foot is a challenge for the perioperative team. Primary closure may not be an option and secondary healing may not be reliable. Therefore, surgery is vital and should be coordinated among a well-functioning multidisciplinary team that specializes in caring for patients with diabetes mellitus. Team members must have expertise in reconstructive surgery to ensure adequate wound healing. This article emphasizes the appropriate timing and staging of surgery, discusses the most common plastic surgery techniques, and underscores the importance of a team approach in the management of diabetic foot wounds. PMID- 18512305 TI - Surgical treatment of Charcot neuropathy. AB - Charcot neuroarthropathy, a chronic progressive destruction of joint integrity, is believed to result from a disturbance in pain and proprioceptive sensation. It is most commonly treated in patients with uncontrolled diabetes mellitus and dense peripheral neuropathy. Prevention, early diagnosis, and early treatment are key to a patient's successful outcome. Educating the patient is paramount to avoid further complications and subsequent amputations. This article describes the pathophysiology, staging, surgical treatment, and natural course of Charcot neuroarthropathy. PMID- 18512306 TI - [Diagnostic tests for myelodysplastic syndromes: current standards and recommended procedures]. PMID- 18512307 TI - [Studying of clinical and laboratory features of chronic eosinophilic leukemias /hypereosinophilic syndrome]. AB - OBJECTIVE: To investigate the clinical and laboratory features of chronic eosinophilic leukemias (CEL) and hypereosinophilic syndrome (HES). METHODS: The clinical manifestations, laboratory parameters were retrospectively analyzed in 20 patients with HES/CEL. Detection of the FIP1L1-PDGFRA fusion gene was performed by nested RT-PCR. JAK2 V617F mutation screening was processed through allele-specific PCR combined with sequence analysis. PCR-RFLP was used to discriminate homozygous from heterozygous mutation patterns. TCR gamma rearrangement was detected by PCR. RESULTS: Of the 20 patients, 19 were males and one female, with a median age of 33 (20 to 57) years. The FIP1L1-PDGFRA fusion gene positivity in bone marrow mononuclear cells in 12 cases was identified. All the breakpoints were identified by direct sequencing of cloned RT-PCR products in FIP1L1 intron 10 - 12 and in PDGFRA exon 12. In CEL the most common involved organs were lungs, heart and nervous system. Splenomegaly was significantly more frequent in CEL than in HES (92.5% vs 42.5%, P = 0.031). Anemia and myelofibrosis were common in CEL. There was no significant difference in circulating absolute eosinophil, leukocyte, platelet counts, hemoglobin level and percentages of eosinophil and blast cell in bone marrow between CEL and HES. The morphological abnormalities of eosinophils on bone marrow smear were easily found in CEL, including hypogranularity, and cytoplasmic vacuolization, increased basophilic granule. One patient with HES was found to have heterozygous JAK2 V617F mutation. Six patients had TCR gamma rearrangement, including 4 CEL and 2 HES. CONCLUSIONS: (1) There is a male predominance in HES/CEL, and the median age was in the thirties. (2) The most common involved organs in CEL were lung, heart and nervous system. Bone marrow morphology might be of a little help in diagnosis of CEL. (3) JAK2 V617F may be involved in the pathogenesis of HES. (4) Patients with CEL carried the FIP1L1-PDGFRA fusion gene and TCR gamma rearrangement concurrently, their relationship warrants further study. PMID- 18512308 TI - [HAA regimen as induction chemotherapy for newly diagnosed acute myelogenous leukemia]. AB - OBJECTIVE: To analyse the outcome of newly diagnosed adult acute myeloid leukemia (AML) patients treated with HAA (homoharringtonine, cytarabine and aclarubicin) regimen and explore the efficacy and safety of this regimen. METHODS: Eighty patients were treated with HAA regimen. The complete remission (CR) rate was observed. Kaplan-Meier method was used to estimate relapse free survival (RFS) rate and the differences were compared with 2-sided log-rank test. RESULTS: Of the 80 patients, 65 (81%) attained CR and the CR rate after the first course of induction was 75%. For the CR patients, the median follow-up was 26 (2 -69) months, and the estimated 3-year overall survival (OS) rate was 51% and the estimated 3-year RFS was 53%. For the AML-M5 and AML-M /M2 patients the CR rate was 74% and 87% and 3 year RFS of CR patients was 75% and 37%, respectively. The CR rate of 100%, 83% and 20% was achieved in patients with favorable, intermediate and unfavorable cytogenetics, respectively. The 3 year OS for favorable and intermediate group was 76% and 50% respectively. The median survival time of unfavorable group was only 6 months. CONCLUSION: HAA regimen is a safe, efficacious, and well-tolerable induction therapy for newly diagnosed AML. PMID- 18512309 TI - [Efficacy and safety of imatinib in treatment of 151 chronic myeloid leukemia patients]. AB - OBJECTIVE: To evaluate the safety and efficacy of imatinib in treatment of chronic myeloid leukemia (CML) patients. METHODS: From December 2003 to March 2007, 151 patients entered Glivec International Patient Assistance Program (GIPAP) in our center and received imatinib therapy. The overall and progression free survival, hematologic, cytogenetic and molecular response, and adverse events were evaluated. The factors associated with outcome of imatinib therapy were also analysed. RESULTS: One hundred and forty-two patients were evaluable with a median follow-up duration of 21.5 (6 -78) months. (1) The rate of cumulative complete hematologic response (CHR), major cytogenetic response (MCyR), complete cytogenetic response (CCyR) and complete molecular response (CMoR) in chronic phase (CP) CML patients were 96.9%, 82.6%, 76.1% and 29.4%, respectively. These rates were significantly higher in patients with CP than in those with accelerated phase (AP) and blast crisis (BC) (P < 0.0001). (2) The overall survival (OS) rates at 1, 2 and 3 year were 100%, (97.3 +/- 1.9)% and (95.8 +/- 2.4)% for CP patients, they were (84.7 +/- 8.2)%, (77.0 +/- 10.4)% and (69.3 +/- 11.9)% for AP patients, and (62.9 +/- 8.9)%, (41.9 +/- 9.2)% and (28.5 +/- 9.1)% for BC patients, respectively (P < 0.0001). The progression-free survival (PFS) rates at 1, 2 and 3 year were (98.9 +/- 1.1)%, (93.9 +/- 2.7)%, (93.9 +/- 2.7)% for CP patients, (68.9 +/- 10.6)%, (61.3 +/- 11.9)%, (61.3 +/- 11.9)% for AP patients, (36.4 +/- 8.8)%, (25.4 +/- 8.1)%, (10.1 +/- 8.2)% (P < 0.0001) for BC patients respectively. (3) Among 92 CP patients, the rates of MCyR and CCyR in newly diagnosed patients were significantly higher than those in interferon therapy failure patients (P = 0.015, P = 0.010). Patients obtained CCyR at 12 months after the initiation of imatinib treatment were associated with longer PFS (P = 0.0099). According to the Sokal scoring system, the rates of MCyR and CCyR in low-risk patients were significantly higher than those in intermediate-risk and high-risk patients (P = 0.0013, P = 0.0024). Sokal score was also significantly associated with disease progression (P = 0.0467). (4) The adverse events of imatinib were moderate and tolerable. CONCLUSIONS: Treatment of CML patients in CP with imatinib can induce high hematologic, cytogenetic and molecular response and overall survival, but can not do satisfactorily for patients in AP and BC. PMID- 18512310 TI - [Analysis of long-term treatment outcome and related factors in 95 chronic myeloid leukemia patients treated with imatinib]. AB - OBJECTIVE: To investigate the efficacy of imatinib in the treatment of chronic myeloid leukemia (CML) and analyse the treatment outcome and related factors. METHODS: Ninety five CML patients were treated with imatinib in our hospital from May 2002 to May 2006. The outcomes and related factors were analysed. RESULTS: (1) One year after therapy, there were 95.5% of chronic phase (CP) patients achieved complete hematologic response (CHR). Fifty-two patients with complete cytogenetic dates were divided into primary-therapy group (n = 19) and secondary therapy group (n = 33). The major cytogenetic responses (MCyR) at 6-, 12-, 18-, 24- and 30-months after therapy for the former group were 84.2%, 84.2%, 89.5%, 89.5% and 94.7%, and for the latter group were 36.4%, 39.4%, 39.4%, 39.4% and 39.4%, respectively (P < 0.01). The expected survival at 12-, 24-, 36- and 50 month after imatinib treatment for CP group was (98.1 +/-1.9)%, (87.8 +/- 7.1)%, (81.9 +/- 8.7)% and (81.9 +/- 8.7)%, respectively. (2) Twelve month after therapy, there are 70% of accelerated phase (AP) patients achieve CHR and 10% get MCyR. The expected survival at 12-, 24- and 36-month after imatinib treatment for AP group was (63.0 +/- 17.7)%, (15.8 +/- 14.3)% and (15.8 +/- 14.3)%, respectively. (3) Six month after therapy, 57.9% of blast crisis (BC) patients achieve CHR, with the expected survival at 12- and 24-month of (40.6 +/- 12.3)% and 0, respectively. (4) COX analysis CP group indicated that imatinib therapy administered for previously untreated was an independent favorable prognostic factor. Conclusion (1) Imatinib as a primary treatment for CP CML can significantly improve the survival time as compared with that AP or BC patients or with that used in previously treated patients. (2) Imatinib could induce hematologic, even cytogenetic response to a certain extent, in CP or BC patients and prolong the survival time. PMID- 18512311 TI - [The immunophenotypic analysis of CD7+ and (or) CD56+ acute myeloid leukemic stem cells and its applications in minimal residual disease detection]. AB - OBJECTIVE: To explore the immunophenotypic characteristics of CD7 and/or CD56 positive acute myeloid leukemic stem cells, and the relationship between minimal residual disease (MRD) and the leukemic stem cells (LSC). METHODS: The immunophenotype of leukemia cells from 51 CD34+ CD38+ CD7+ and/or CD34+ CD38+ CD56+ acute myeloid leukemia (AML) patients (exclude M3) at diagnosis was analyzed by using 4 - 6 panels of 4 color antibodies, and cells from 28 normal bone marrow (NBM) samples were served as control. The expression of CD7 and CD56 in the CD34+ CD38+ subpopulation was used as a leukemic cell marker for monitoring MRD of 53 samples from 26 CD7+ and (or) CD56+ patients. RESULTS: In CD7+ and/or CD56+ AML patients at diagnosis, the average positivity of CD7 in CD34+ CD38+ subpopulation and CD34+ CD38- Lin- stem cells subpopulation was (77.39 +/- 20.71)% and (44.57 +/- 22.70)%, and that of CD56 was (56.71 +/- 32.56)% and (33.51 +/- 29.64)%, respectively, all significantly higher than that of NBM (P < 0.01 and P < 0.05). Compared with that of NBM, the expression of CD90 in AML patients was significantly lower in the CD34+ CD38- Lin- subpopulation (P < 0.01), the expression of CD123 was significantly higher than NBM (P < 0.01), and the expression of CD117 was no significant difference (P > 0.05). In follow up of CD7+ and (or) CD56+ patients, the expression rate of CD7 and (or) CD56 in the CD34+ CD38- Lin- subpopulation MRD+ group was significantly higher than that in the MRD- group. The actual rate for CD7 was 71% (15/21) and 16% (4/25) (P = 0.001), and its relative rate was 81% (17/ 21) and 24% (6/25) (P = 0.000), respectively. The actual rate of CD56 is 100% (4/4) and 12% (3/25) (P = 0.001), and its relative rate was 75% (3/4) and 20% (5/25) (P = 0.031), respectively. A high CD7+ CD34+ CD38- Lin- subpopulation frequency at diagnosis in CD7+ AML patients predicted a high frequency of positive MRD in later detection (P < 0.05). CONCLUSIONS: CD7 and CD56 are expressed on the stem cells in CD7+ and/or CD56+ AMLs and a high frequency of CD7 and CD56 in the CD34+ CD38- Lin- stem cell subpopulation predicts a high frequency of positive MRD in later detection. PMID- 18512312 TI - [Comparison of polyethylene glycol conjugated asparaginase and L-asparaginase for treatment of childhood acute lymphoblastic leukemia]. AB - OBJECTIVE: To evaluate efficacy and side-effect of polyethylene glycol conjugated asparaginase (PEG-Asp) containing VDPAP regimen in newly diagnosed childhood acute lymphoblastic leukemia (ALL). METHODS: One hundred and thirty five children with newly diagnosed ALL were randomized assignments to receive PEG-Asp containing VDPAP regimen or L-asparaginase containing VDLP regimen as control for induction therapy. The VDPAP regimen consisted of daunomycin on day 1, 2, prednisone for 28 days, vincristine once a week for 4 weeks, and PEG-Asp(2500 IU/m2) intramuscularly once every two weeks for 4 weeks. In the VDLP regimen, the PEG-Asp was substituted by L-asparaginase (6000 IU/m2, three times a week for 2 weeks). The pharmacokinetics of PEG-Asp was investigated in 11 patients in VDPAP group. RESULTS: The complete remission (CR) rates were 84.6% and 89.4%, total response rates were 92.3% and 93.9% (P < 0.05, respectively), and the incidence of adverse effects was 95.6% and 98.5% (P > 0.05) in the VDPAP and VDLP group, respectively. The major adverse effect was allergic response (4.4% and 5.9% in the VDPAP and VDLP group, respectively) and others included coagulopathy and gastrointestinal symptoms. Pharmacokinetics showed that the half-life of PEG-Asp was (170 +/- 45) hours [about (7 +/- 2) days], the serum PEG-Asp activity levels > 0.1 IU/ml throughout induction. CONCLUSION: The effectiveness of PEG-Asp and L asparaginase for treatment of pediatric ALL is similar and no difference in adverse effect. The advantage of PEG-Asp is more prolonged effect and convenient. PMID- 18512313 TI - [Targeted blockage of RNA binding protein E2 by decoy RNA induces the granulocytic differentiation of K562 cells]. AB - OBJECTIVE: To use a decoy RNA targeted blockage of the RNA binding protein E2 (hnRNP E2) resulting in the CCAAT/enhancer-binding protein alpha (C/EBP alpha) gene's abnormal translation and investigate its effect on the granulocytic differentiation of K562 cells and the probable molecular mechanism. METHODS: The hnRNP E2 decoy RNA expression plasmid was constructed and transfected into K562 cells with cationic liposome, and stable expression cells were obtained by G418 selection. The changes of C/EBP alpha and granulocyte colony-stimulating factor receptor (G-CSFR) gene expression were detected by RT-PCR and Western blot. The morphologic changes were observed after Wright-Giemsa staining. The expression of granulocytic differentiation antigens CD13 and CD15 was studied by immunocytochemistry. RESULTS: The stably expressed pG cells were obtained. Its C/EBP alpha mRNA level remained unchanged, while 42kD-C/EBP alpha protein expression was increased by (49.7 +/- 5.5)% (P < 0.05); and G-CSFR mRNA was increased by (42.1 +/- 3.6)% (P < .05), and its protein was increased by (37.4 +/ 6.2)% (P < 0.05) compared to that in the K562 control cells. The characteristics of polymorphonuclear neutrophils appeared in pG cells and CD13 and CD15 positive cell ratios were (18.7 +/- 2.5)% and (26.3 +/- 2.9)% respectively. CONCLUSIONS: HnRNP E2 decoy RNA can induce granulocytic differentiation of K562 cells, and G CSF promotes this effect. The mechanisms may be that decoy RNA specifically blocks hnRNP E2, hence regulates the translation of C/ EBP alpha mRNA, restores the expression of 42kD-C/EBP alpha, and then up-regulates the expression of G CSFR gene. PMID- 18512314 TI - [The cytotoxicity of CIK/NK cells stimulated by K562-DC fusion vaccines in NOD/SCID mice model for human erythroleukemia]. AB - OBJECTIVES: To study the in vivo efficacy and the safety of cord blood derived CIK/NK cells stimulated by K562-dendritic cells (DC) fusion vaccines in NOD/SCID mice model for human erythroleukemia. METHODS: DC and CIK /NK cells were both derived from cord blood mononuclear cells. DC were fused with inactivated K562 leukemia cell by PEG to produce K562-DC fusion vaccines. K562-DC fusion vaccines were co-cultured with CIK/NK cells to prepare K562-DC fusion vaccine stimulated CIK/NK cells. NOD/SCID mice were inoculated with 1 x 10(6) K562 cells. 24 hours later, 1 x 10(7) vaccines stimulated CIK/ NK cells and 1 x 10(7) CIK/NK cells were transfused into the NOD/SCID mice. NOD/SCID mice without inoculation of K562 cells were used as control group. CD13 and CD56 positive cells were assayed by flow cytometry. RESULTS: All the leukemia NOD/SCID mice without therapy died within 39 days, tumor was found in 5 of 8 mice. One of 8 leukemia mice treated with K562-DC fusion vaccines stimulated CIK/NK cells died at the 65th day, the anti-tumor response rate was 87.5%. Two of the leukemia mice treated with CIK/NK cells died at the 56th and 65th day respectively, the anti-tumor response rate was 75%. There was no significant difference in survival time between these two groups, and both survivals were longer than that of the control group. There was no significant difference in CD13 positive cells in the survival mice between these two groups, and both of that were less than that of the control mice. There was no significant difference in CD56 positive cells between the two treated groups and the control group. CONCLUSIONS: Cord blood derived CIK/ NK cells stimulated by inactivated tumor cells retain the cytotoxicity and do not develop tumor in vivo. PMID- 18512315 TI - [Relationship between the long type PML-RAR alpha and the prognosis of patients with acute promyelocytic leukemia]. AB - OBJECTIVE: To explore the relationship between long (L) type PML-RAR alpha fusion gene and the prognosis of patients with acute promyelocytic leukemia (APL). METHODS: PML-RAR alpha fusion gene was detected by RT-PCR in 33 APL patients. The optical density of three bands including E5 (+) E6 (+) 636 bp (no deletion), E5 ( ) E6 (+) 492 bp (exon 5 deleted) and E5 (-) E6 (-) 232 bp (both exon 5 and exon 6 deleted) was measured by a UVP analysis system and their relative proportions were calculated. The relative expression level of each splicing band, initial WBC count and age were statistically (for single and multi-factor analysis) analyzed with prognosis. RESULTS: The relative expressions of E5 (-) E6 (+) and E5 (-) E6 (-) in death group were obviously different from that in first complete remission (CR1) group (P < 0.01), but do not for E5 (+) E6 (+) (P > 0.05). The relative expression levels of E5 (-) E6 (+), E5 (-) E6 (-) and E5 (+) E6 (+) were 0.23 +/- 0.12, 0.58 +/- 0.18, 0.20 +/- 0.09 in death group, and 0.45 +/- 0.16, 0.23 +/- 0.12, 0.31 +/- 0.16 in CR1 group, respectively. Initial WBC count and age was no difference between the two groups (P > 0.05). Logistic regression analysis showed that the expression of E5 (-) E6 (+) had no effect on the prognosis (B = 3.475, P = 0.492), but the expression of E5 (-) E6 (-) showed a negative correlation with prognosis (B = -19.660, P = 0.046). CONCLUSIONS: The high expression of E5 (-) E6 (-) is correlated with the poor prognosis for patients with APL. PMID- 18512316 TI - [Study on the pathogenesis of blood cytopenia in patients with systemic lupus erythematosus]. AB - OBJECTIVE: To explore the pathogenetic mechanism of cytopenia in the patients with systemic lupus erythematosus (SLE). METHODS: Twenty-one patients with SLE and 10 healthy controls were enrolled in this study. Bone marrow mononuclear cells (BMMNC) Coombs test was used to determine the autoantibodies and in vitro CFU-E, BFU-E and CFU-GM yields were used to determine the proliferation function of hematopoietic stem/progenitor cell (HSC). RESULTS: Positive results of BMMNC Coombs test were observed in 12 patients with SLE (57.1%), among them, 10 with (58.2%) and 2 without blood cytopenia. The yields of CFU-GM (49 +/- 40), and CFU E (49 +/- 35) in SLE patients without cytopenia were higher than that in normal controls, but lower than that in cytopenia SLE patients. The BFU-E yield in SLE patients (3 +/- 4) was significantly lower than that in normal controls (36 +/- 12). CONCLUSION: There were auto-antibodies on BMMNC in patients with SLE. The blood cytopenia in SLE patients maybe resulted from the destructions of bone marrow hematopoietic cells by the autoantibodies. The capacities of bone marrow HSC in SLE patients for proliferation compensation of hematopoietic cell destruction were not damaged. PMID- 18512317 TI - [Strengthening public health safety control and have a nice service for 2008 Beijing Olympic Games]. PMID- 18512318 TI - [Research of public health risks and tackle strategies for 2008 Beijing Olympic Games]. AB - OBJECTIVE: To identify and evaluate the risks of serious public health events for 29th the Olympic Games and 13th Paralympic in order to offer scientific bases for security public health of Olympic Games. METHODS: Use Delphi experts refer, Level analysis, Risk assessment and Hazard analysis and critical control points, to research public health risks for 2008 Beijing Olympic Games. RESULTS: From analysis, we concluded 45 kinds of public health risk consisting in 5 areas and made sure its risk level: 5 kinds of Extreme risk in 3 areas; 22 kinds of High risk in 5 areas; 10 kinds of Moderate risk in 4 areas and 8 kinds of Low risk in 4 areas. And also we imported the risk management thought to research tackle strategies for 2008 Beijing Olympic Games. CONCLUSIONS: It might offer scientific bases for security public health of Olympic Games by identification and risk assessment of Serious Public Heath Events for the 29th Olympic Games. PMID- 18512319 TI - [A study of risk assessment indicators system of infectious disease event for 2008 Beijing Olympic Games]. AB - OBJECTIVE: To investigate and establish a risk assessment indicator system of infectious disease event for 2008 Beijing Olympic Games and to assess the risk of infectious disease epidemic on 2008 Beijing Olympic Games, as to offering scientific bases for security public health of Olympic Games. METHODS: Risk assessment subject discussion group was constituted to collect all sorts of data of infectious disease and the risk assessment and synthesis were analyzed. Then the expert's opinions were consulted to confirm the entered indicators and the judged content of indicators. RESULTS: A preliminary risk assessment indicator system of infectious disease events of 2008 Beijing Olympic Games was established. The system mainly include six first level indicators and twenty judge indicators altogether. CONCLUSIONS: The risk assessment indicators system of infectious disease event for 2008 Beijing Olympic Games should be considered as all the risk factors and general application characteristics. It might be used in any security procedure of important function and risk management. PMID- 18512320 TI - [Effects of aluminum on the integrity of blood brain barrier in juvenile rats]. AB - OBJECTIVE: To investigate the effects of aluminum on the integrity of blood brain barrier in juvenile rats. METHODS: The 40-day old Sprague-Dawley (SD) rats were exposed to aluminium chloride by intraperitoneal injection, at a dose of Al3+ 0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, respectively. Morris water amaze system was used to test the learning and memory ability. The Evans blue content in brain was analyzed after injection. The ultrastructure's change of the blood brain barrier (BBB) was observed with transmission electron microscope. The expression of occluding protein in BBB was determined by Western blot method. RESULTS: As compared with control group, the permeability of BBB in mid-level Al and high level Al was enhanced (P <0.01), the expression of occluding protein was descended (P <0.01). The ultrastructures of the BBB were changed. No differences between every group on learning and memory ability (P>0.05). CONCLUSIONS: Short time and low dose of Al might not change the ability of learning and memory in juvenile rats, however the permeability and ultrastructures of the BBB might be significantly changed. PMID- 18512321 TI - [Effects of maternal high protein diet on uncoupling protein and carnitine palmityl transferase 1 in offspring of rats]. AB - OBJECTIVE: To investigate the effects of maternal nutritional manipulation on fetal mRNA abundance of uncoupling protein UCP2, UCP3 and carnitine palmityl transferase 1 (CPT1), and find out an optimal maternal diet and targets for pharmacological prevention and treatment of obesity. METHODS: Wistar pregnant rats were assigned to two groups which received a standard diet (SD) and a high protein diet (HPD) during pregnancy, respectively. After delivery, the male offspring were assigned to control group (CON) and high protein group (HP) according to their maternal diet, which were suckled by dams that received SD during pregnancy. Offspring were fed with SD from weaning (week 3) to week 8. Then CON were allocated to two groups: CON (SD during the whole experiment); HFCON (high fat control). HFCON and HP group rats were fed with high-fat diet (HFD) for 6 wk to induce obesity. At 0, 3, 8 and 14 wk of age, blood and tissue were collected for analyzing blood fat and abundance of UCP2, 3 and CPT1 mRNA. RESULTS: In HP body weight and TG were decreased after weaning (F = 4.589, P = 0.039; F = 27.001, P = 0.000) and HFD (F = 16.076, P = 0.00; F = 71.518, P = 0.000). Obesity rates were significantly decreased in HP after HFD (chi2 = 8.076, P = 0.004). The abundance of UCP3 and CPT1 mRNA was persistently higher in HP than in CON or HFCON, and the abundance of UCP2 mRNA was also persistently higher than in CON or HFCON after weaning. Moreover the abundance of CPT1 mRNA was significantly increased after weaning and HFD compared with that after SD, the abundance of UCP2, UCP3 mRNA was also increased after HFD compared with that after SD. CONCLUSIONS: Increasing protein intake during pregnancy might prevent offspring from HFD-induced obesity in adult, moreover might increase offspring the expression of UCP2, UCP3 and CPT1 mRNA. UCP2, UCP3 and CPT1 might participate in prevention and treatment of obesity by mediating fatty acid oxidation. PMID- 18512322 TI - [Effect of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin on the expression of AhR and TGF-alpha]. AB - OBJECTIVE: To study HaCaT-keratinocyte cell lines, a chosen model of human epidermis in an attempt to analyze the mRNA expression of AhR and TGF-alpha induced by TCDD METHODS: Semi-quantitative reverse transcription PCR-technique was used for assaying the relative levels of AhR and TGF-alpha mRNA of HaCaT cells during the proliferation period when the cells were cultured for 24 hours. RESULTS: Relative level of the AhR-transcripts (corrected with beta-actin) decreased with the elevated concentration of TCDD and the relevant coefficient between the proliferation rate and concentration was -0.548, and the differences among all groups were significant (F =4.124, P =0.021). The vehicle control was respectively compared with 7 x 10(-10) mol/L (0.0620 +/- 0.0085) and 7 x 10(-9) mol/L (0.0518 +/- 0.0194) group, significantly different from the control group (0.1138 +/- 0.0227) (t = -3.48, P <0.05; t = -4.17, P <0.01), the expression amount being 55% and 45% of the control. Relative levels of TGF-alpha mRNA tended to increase with the elevated concentration with the significant coefficient of 0.695 (P < 0.01), and the differences among all groups were significant (F = 15.789, P =0.000). In two higher concentration group 7 x 10(-10) mol/L (0.1474 +/ 0.0390) and 7 x 10 (-9) mol/L (0.2133 +/- 0.0364), their relative expression amount of TGF-alpha mRNA was 2.6-fold, 3.8-fold of the control group (0.0561 +/- 0.0100) respectively. Further analysis for the relevant relationship between the amounts of the AhR mRNA and TGF- alpha mRNA showed a highly negative correlation, the coefficient being - 0.561 (P <0.05). CONCLUSIONS: TCDD down-regulate the expression of AhR and up-regulate the expression of TGF-alpha. A strong negative correlation between AhR and TGF-alpha expression is found. PMID- 18512323 TI - [Characteristic of the neurobehavioral functional changes in coke oven workers]. AB - OBJECTIVE: To explore the effect of polycyclic aromatic hydrocarbons on the neurobehavioral function of coke oven workers. METHODS: 200 healthy adult male coke oven workers were selected from a coke plant of a state-owned steel enterprise in Taiyuan City. 88 controls occupationally unexposed to polycyclic aromatic hydrocarbons (PAHs) were selected from the same enterprise. All the subjects participated in this investigation voluntarily in their consent. Concentration of B(a)P in the working environment was monitored by High Performance Liquid Chromatography (HPLC). Urine samples were sampled immediately after working shifts. The level of urinary 1-hydroxypyrene was determined by HPLC. General information of workers correlated with the investigation was collected in a questionnaire according to the same criteria by well-trained investigators. Neurobehavioral core test battery (NCTB) recommended WHO was performed on coke oven workers and controls to test the neurobehavioral changes and the mood state. RESULTS: the concentration of B(a)P at oven bottom,oven side and oven top were 0.0195 microg/m3, 0.186 microg/m3 and 1.624 microg/m3 respectively, that at oven side and oven top being higher than the one stipulated by the occupational hygiene criterion. Urinary 1-hydroxypyrene was significantly different between the exposure group (3.42 +/- 0.98 micromol/mol creatinine) and control group (2.75 +/- 1.09 micromol/mol creatinine). No significant differences were found between exposure group and control group of age, working years, smoking, drinking and unhealthy food consumption; however, compared to the controls, the scores of total digital span, the forward digital span, and right dotting in the coke oven workers were lower, but that of total dotting was higher, with a statistical significance. According to urinary 1-hydroxypyrene concentration, all the subjects were divided into three groups. (<3.10 micromol/mol creatinine, 3.10 micromol/mol creatinine, >3.87 micromol/mol creatinine). Significant differences of the total digital span, the forward digital span, backward digital span, digit symbol and Benton visual retentions existed in different urinary 1-hydroxypyrene concentration groups and showed a dose-response tendency. Results of multiple stepwise regression analysis and correlation analysis showed that the level of urinary 1-hydroxypyrene affected memory and perception of coke oven workers and negative correlations between the level of urinary 1-hydroxypyrene and changes in neurobehavioral function were found. CONCLUSION: PAHs mainly causes decrease of memory and perception in coke oven workers. PMID- 18512324 TI - [Comparative study on the acute toxicity of pulmonary caused by nanosized and microsized powders of silicon dioxide]. AB - OBJECTIVE: To compare the acute pulmonary toxicities of nanosized and microsized silicon dioxide particles. METHODS: All 125 healthy male Wistar rats were divided into 25 groups randomly according to the weight. Experimental animals were exposed to microsized SiO2 at the doses of 100 mg/m3 (group A) and 300 mg/m3 (group B), and to the nanosized SiO2 at the same dose levels (group A' and B') by inhalation for 2 hours. Compositions in bronchoalveolar lavage fluids (BALF) and contents of hydroxyproline in blood sera and lung tissues were detected and then compared at 6, 12, 24, 48, 72 hours after administration. RESULTS: The total cellular score (TCS) in BALF of group A'[(55.00 +/- 8.30) x 10(4)/ ml] and B'[(52.50 +/- 9.02) x 10(4)/ml] at 6 hours were significantly higher than those in control groups [(34.88 +/- 12.53) x 10(4)/ml]; TCS in BALF of group A' [(55.00 +/- 8.30) x 10(4)/ml]at 6 hours and group A' [(39.75 +/- 12.08) x 10(4)/ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(32.38 +/- 13.07) x 10(4)/ml, (24.13 +/- 10.97) x 10(4)/ml) ]; total protein (TPr) in BALF of group A' [(0.34 +/- 0.09)g/L] and B' [(0.38 +/- 0.16) g/L] at 48 hours were significantly higher than those in isodose group of microsized SiO2 [(0.20 +/- 0.07) g/L, (0.21 +/- 0.05) g/L]. Lactate dehydrogenase (LDH) in BALF of group A' [(1.66 +/- 0.22) x 10(3) U/L] at 72 hours were significantly higher than those in isodose group of microsized SiO2 [(1.38 +/- 0.17) x 10(3) U/L]. Alkaline phosphatase (AKP) in BALF of group B' [(5.14 +/- 1.47) U/100 ml] at 6 hours and group B' [(5.86 +/- 2.41) U/100 ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(3.64 +/- 0.36) U/100 ml, (3.30 +/- 2.19) U/100 ml]. Hydroxyproline (HyP) in tissues of lung of group A' [(0.532 +/- 0.053) microg/mg, (0.484 +/- 0.046) microg/mg, (0.591 +/- 0.096) microg/mg, (0.551 +/- 0.084) microg/mg] at 6, 12, 48, 72 hours and group B' [(0.508 +/- 0.081) microg/mg, (0.565 +/- 0.053) microg/mg ] at 12, 72 hours were significantly higher than those in isodose group of microsized SiO2 [(0.345 +/- 0.074) microg/mg, (0.368 +/- 0.095) microg/mg, (0.431 +/- 0.036) microg/mg, (0.399 +/- 0.080) microg/mg, (0.396 +/- 0.039) microg/mg, (0.465 +/- 0.062) microg/mg]. CONCLUSION: Nanosized and microsized SiO2 should have some differences on acute pulmonary toxicities in our experiment condition. PMID- 18512325 TI - [Application of pulse-field gel electrophoresis analysis in source-tracking of food-borne disease caused by Vibrio parahaemolyticus]. AB - OBJECTIVE: To apply pulse-field gel electrophoresis analysis(PFGE) in analysing a case of food poisoning caused by Vibrio parahaemolyticus. METHODS: PFGE using restriction enzyme Not I was employed in molecular subtyping of thirty strains of V. parahaemolyticus isolated from a case of food poisoning in Guangzhou city and PFGE patterns were analyzed by using BioNumerics Version 4.0 software to perform cluster analysis. Pattern profiles were compared by using the Dice coefficient and unweighted pair group method with arithmetic averages (UPGMA). RESULTS: Thirty strains were of the same type of pulsotype. CONCLUSIONS: Molecular subtyping by PFGE might disclose the epidemiological relationships of the strains from humans, food and the environment, giving a strong molecular epidemiological evidence and a support for the source-tracking of outbreak events. PMID- 18512326 TI - [Effects of garlic oil combined with resveratrol on inducting of apoptosis and expression of Fas, bcl-2 and bax in human gastric cancer cell line]. AB - OBJECTIVE: To explore the effects of garlic oil combined with resveratrol on the apoptosis and expression of Fas, bcl-2 and bax in human gastric cancer cell line MGC-803. METHODS: The experiment included three groups which were the control group, the combined medicine group 1 (including 25 microg/ml oil garlic and 25 microg/ml resveratrol) and the combined medicine group 2 (including 50 microg/ml oil garlic and 50 microg/ml resveratrol). The apoptosis of cell was examined by DNA gel electrophoresis and flow cytometry for annexin v; the expression of Fas was determined by flow cytometry at the 24th hour after the treatment; the mRNA expression of bcl-2 and Bax gene were measured by RT-PCR method at the 24th ang 48th after treatment, respectively. RESULTS: The garlic oil combined with resveratrol induced cell apoptosis markedly at the 24th after the treatment The protein expression of Fas in the combined medicine groups was 10.59% and 14.16% respectively. As compared with the control group (5.27%), the statistical significance was obvious. The mRNA level of Bax was elevated significantly, however the mRNA expression of bcl-2 was decreased at the 24th and 48th after the treatment. CONCLUSIONS: The garlic oil combined with the resveratrol might obviously induce the apoptosis of gastric cancer cell line MGC-803 which be involved in increasing the expression of Fas protein and bax gene and decreasing the expression of bcl-2 gene at the same time. PMID- 18512327 TI - [Viral load test conducive to excluding negative subjects from suspects in HIV antibody detections]. AB - OBJECTIVE: To study whether plasma viral load testing is helpful to exclude ones free from Human immunodeficiency virus (HIV) infections from suspects in HIV antibody detections. METHODS: 19 Specimens, which showed disconcordant results of the two HIV EIA testing (S/CO < 6) and indeterminated results of Western blot (WB) test, were selected. Viral load of the specimens were detected. A six-month follow up survey in detecting HIV antibody was conducted in these subjects. RESULTS: None of these 19 cases was observed to be positive HIV viral loads and there was no any progress in WB bands development during the follow-up period. The possibility of HIV infection could be excluded. CONCLUSION: When the specimens react with very low intensity in both EIA and WB, negative viral load result is conducive to exclude negative subjects from suspects in HIV antibody detections. PMID- 18512328 TI - Cloning of an ovule specific promoter from Arabidopsis thaliana and expression of beta-glucuronidase. AB - Tissue specific expression of transgenes in plant species has several advantages over constitutive expression. Identification of ovule specific promoters would be useful in genetic engineering of plants with a variety of desirable traits such as genetically engineered parthenocarpy, female sterile plants or seedless fruits. Relative inaccessibility and difficulty in harvesting adequate amounts of tissue at known developmental stages has impeded the progress in cloning of promoters involved in ovule development. In the present study an ovule specific promoter was cloned from Arabidopsis AGL11 gene and used to express GUS (beta glucuronidase) gene in transgenic Arabidopsis. Histochemical staining of GUS appeared in the center of young ovary (ovules), but no detectable GUS activity was observed in vegetative plant tissues, sepals, petals and androecium. AGL11 gene promoter can be useful to modify the developmental path of plants by expressing either plant hormones or lethal genes for agronomic purpose. PMID- 18512329 TI - Inhibition of MAO and GABA: probable mechanisms for antidepressant-like activity of Nardostachys jatamansi DC. in mice. AB - Ethanolic extract (100, 200 and 400 mg/kg, po) of N. jatamansi administered for 14 successive days to Swiss young albino mice (either sex) produced significant antidepressant-like effect in both tail suspension and forced swim tests. The efficacy of the extract was found to be comparable to imipramine (15 mg/kg, po) and sertraline (20 mg/kg, po). Ethanolic extract (200 mg/kg, po) did not show any significant change on locomotor activity of mice as compared to control; hence it did not produce any motor effects. Further, the extract decreased the whole brain MAO-A and MAO-B activities as compared tocontrol, thus increased the levels of monoamines. The antidepressant effect of the extract was also significantly reversed by pretreatment of animals with baclofen (GABAB agonist); when tested in tail suspension test. The results suggested that the antidepressant-like effect of the extract may also be due to interaction with GABAB receptors, resulting in decrease in the levels of GABA in mouse brain. Thus, the extract may have potential therapeutic value for the management of mental depression. PMID- 18512330 TI - Nimodipine is more effective than nifedipine in attenuating morphine tolerance on chronic co-administration in the rat tail-flick test. AB - Opioids, when co-administered with L-type calcium channel blockers (L-CCBs) show morphine like higher antinociceptive effect. This antinociceptive effect has been further investigated using a different experimental paradigm. The effect of two different L-CCBs (nifedipine and nimodipine) on morphine-induced antinociception was studied by the tail-flick test (40 min after morphine administration) in adult Wistar rats. A fixed-dose of nimodipine or nifedipine (2 mg/kg, once daily) was combined with a fixed dose of morphine (10 mg/kg, twice daily) for 10 days. Co-administration of L-CCBs significantly increased the antinociceptive effect of morphine, even 12 hr after administration. Also, nimodipine was more effective than nifedipine. Nimodipine was further studied using a higher and escalating doses of morphine (20-30 mg/kg twice daily for 14 days). Nimodipine increased the antinociceptive effect of morphine in the latter part of the study (days nine to fourteen) though significant difference was observed on 11th evening and 12th morning. No obvious adverse effects were observed in the present study. The results show for the first time that nimodipine is more effective than nifedipine and that these L-CCBs continue to be effective, even 12 hr after administration in the tail-flick test. PMID- 18512331 TI - Bradycardia induced by Mesobuthus tamulus scorpion venom involves muscarinic receptor-G-protein-coupled cell signaling pathways. AB - Indian red scorpion (Mesobuthus tamulus; MBT) envenomation produces various cardio-respiratory abnormalities including cardiac dysrhythmias. The underlying cell signaling pathways for the cardiac dysrhythmias produced by MBT venom are not known. The present study was therefore conducted to delineate the second messenger signaling pathways involved in MBT venom-induced atrial rhythm changes. The effects of venom and various antagonists were examined on spontaneously beating rat right atrial preparations in vitro. The MBT-venom produced an increase (35%), a decrease (45%) and again an increase (50%) in rate at 0.03, 0.3 and 3.0 microg/ml of venom, respectively. On the other hand, force of contraction exhibited a concentration-dependent rise (up to 40%) at all concentrations of venom. Pretreatment with atropine (0.3 microM) blocked the decrease in atrial rate at 0.3 microg/ml concentration of venom while no such blockade was seen in force of contraction. Submaximal concentration of ACh (0.1 nM) decreased the atrial rate by 25%. In the presence of MBT venom (0.3 microg/ml), ACh-induced fall in atrial rate was enhanced. The venom-induced fall in atrial rate and augmentation of ACh response were blocked by pertussis toxin (PTx; a Gi inhibitor) or methylene blue (a G-cyclase inhibitor). The results indicate that the decrease in atrial rate produced by venom is mediated muscarinic by receptors via Gi-guanylyl cyclase mediated cell signaling pathways. PMID- 18512332 TI - Beneficial effects of Lagenaria siceraria (Mol.) Standley fruit epicarp in animal models. AB - Lagenaria siceraria (Mol.) Standley fruit (bottle gourd), a commonly used vegetable in India is described as cardiotonic and as a general tonic in Ayurveda. Keeping in view the presence of free radical scavenging activity in L. siceraria and involvement of free radicals in the development of various disorders, present studies were designed to evaluate the ethanolic extract of L. siceraria fruit against the disorders where free radicals play a major role in pathogenesis. The extract was found effective as hepatoprotective, antioxidant, antihyperglycemic, immunomodulatory, antihyperlipidemic and cardiotonic agent. The results showed that the radical scavenging capacity of L. siceraria fruit may be responsible for various biological activities studied. PMID- 18512333 TI - Effects of different factors on immature embryo culture, PLBs differentiation and rapid mass multiplication of Coelogyne suaveolens (Lindl.) Hook. AB - In vitro mass production of C. suaveolens (Lindl.) Hook, an endangered orchid with its snowy white flowers having horticultural potential was accomplished through immature seed culture, and subsequent plant regeneration. The developmental stage of the immature seeds and nutrient media significantly influenced the germination frequency. Seeds at 13 months after pollination cultured on 3% sucrose containing Murashige and Skoog (MS) medium with 9 microM alpha-naphthaleneacetic acid (NAA), and 15% coconut water exhibited 93% germination after 40 days of culture. Upon subculture, the germinated shoots on MS medium with 9 microM BA, 6 microM NAA, 3% casein hydrolysate and 0.1% activated charcoal (AC) yielded >12 shoots per shoot or bud. Addition of AC favoured the enlargement of pseudobulbs and better rooting. The plantlets transferred to community potting mix after in vitro hardening (8-10 wk) displayed 85% survival. PMID- 18512334 TI - Screening for disease resistance in barley cultivars against Bipolaris sorokiniana using callus culture method. AB - Screening for resistant barley genotypes in response to fungal toxin of Bipolaris sorokiniana was assessed on standing barley plants as well as in selected callus lines of the same. For the standing lines tested, those manifesting chlorosis in response to toxin infiltration showed a significantly slower disease progress as compared to the necrotic lines. Also, necrosis in the callus tissues of the susceptible cultivar in MS medium supplemented with different concentrations of the crude toxin was significantly higher than in the callus tissues of the chlorotic lines studied. Similar host response to the toxin in in vitro and field situations open up the possibility of screening barley cultivars for resistance to spot blotch using callus culture as against classical methods of screening in order to increase accuracy and save time and space. PMID- 18512335 TI - Antimicrobial protein from Streptomyces fulvissimus inhibitory to methicillin resistant Staphylococcus aureus. AB - Fermented culture of Streptomyces fulvissimus was found to secrete an antibacterial protein inhibitory to Micrococcus luteus, Bacillus subtilis, Bacillus cereus and methicillin resistant Staphylococcus aureus (MRSA) strains. The extracellular protein from the fermented culture on concentration revealed a high molecular weight peptide of 63kDa on SDS-PAGE gel and the region on gel displayed inhibitory activity against methicillin resistant Staphylococcus aureus. Bioactivity of the extra cellular protein was non-sensitive to proteinase K, alpha chymotrypsin, protease, EDTA (ethylene diamine tetra acetic acid), PMSF (phenyl methyl sulfonyl fluoride) and DMSO (dimethyl sulfoxide) but partially susceptible to amylase and heat. Glycoprotein nature of the proteinaceous compound was confirmed by periodic acid schiffs (PAS) staining. The secretary protein of S. fulvissimus demonstrated a significant activity against MRSA strain. It could be an important source for developing new drugs to control multidrug resistant gram positive bacteria. PMID- 18512336 TI - Deaths: final data for 2005. AB - OBJECTIVES: This report presents final 2005 data on U.S. deaths, death rates, life expectancy, infant and maternal mortality, and trends by selected characteristics such as age, sex, Hispanic origin, race, marital status, educational attainment, injury at work, state of residence, and cause of death. METHODS: This report presents descriptive tabulations of information reported on death certificates, which are completed by funeral directors, attending physicians, medical examiners, and coroners. The original records are filed in the state registration offices. Statistical information is compiled into a national database through the Vital Statistics Cooperative Program of the Centers for Disease Control and Prevention's National Center for Health Statistics (NCHS). Causes of death are processed in accordance with the International Classification of Diseases, Tenth Revision (ICD-10). RESULTS: In 2005, a total of 2,448,017 deaths were reported in the United States. The age-adjusted death rate was 798.8 deaths per 100,000 standard population, representing a decrease of 0.2 percent from the 2004 rate and a record low historical figure. Life expectancy at birth remained the same as that in 2004-77.8 years. Age-specific death rates decreased for the age group 65-74 years but increased for the age groups 15-24 years, 25-34 years, and 45-54 years. The 15 leading causes of death in 2005 remained the same as in 2004. Heart disease and cancer continued to be the leading and second leading causes of death, together accounting for almost one half of all deaths. The infant mortality rate in 2005 was 6.87 deaths per 1,000 live births. CONCLUSIONS: Generally, mortality patterns in 2005, such as the age adjusted death rate declining to a record historical low, were consistent with long-term trends. Life expectancy in 2005 remained the same as that in 2004. PMID- 18512337 TI - Molecular mechanisms of parasite invasion. Preface. PMID- 18512338 TI - Current and emerging approaches to studying invasion in apicomplexan parasites. AB - In this chapter, we outline the tools and techniques available to study the process of host cell invasion by apicomplexan parasites and we provide specific examples of how these methods have been used to further our understanding of apicomplexan invasive mechanisms. Throughout the chapter we focus our discussion on Toxoplasmagondii, because T. gondii is the most experimentally accessible model organism for studying apicomplexan invasion (discussed further in the section, "Toxoplasma as a Model Apicomplexan") and more is known about invasion in T. gondii than in any other apicomplexan. PMID- 18512340 TI - Erythrocyte invasion by Plasmodium falciparum: multiple ligand-receptor interactions and phenotypic switching. PMID- 18512339 TI - Microneme proteins in apicomplexans. AB - Microneme secretion supports several key cellular processes including gliding motility, active cell invasion and migration through cells, biological barriers, and tissues. The modular design of microneme proteins enables these molecules to assist each other in folding and passage through the quality control system, accurately target to the micronemes, oligimerizing with other parasite proteins, and engaging a variety of host receptors for migration and cell invasion. Structural and biochemical analyses of MIC domains is providing new perspectives on how adhesion is regulated and the potentially distinct roles MICs might play in long or short range interactions during parasite attachment and entry. New access to complete genome sequences and ongoing advances in genetic manipulation should provide fertile ground for refining current models and defining exciting new roles for MICs in apicomplexan biology. PMID- 18512341 TI - Role of the gp85/trans-sialidase superfamily of glycoproteins in the interaction of Trypanosoma cruzi with host structures. AB - Invasion of mammalian cells by T. cruzi trypomastigotes is a multi-step and complex process involving several adhesion molecules, signaling events and proteolytic activities. From the blood to the cell target in different tissues the parasite has to interact with different cells and the extracellular matrix (ECM). The review focus on the role of the gp85/ trans-sialidase superfamily members in the interaction of the parasite with the host cell, particularly with ECM components, with emphasis on the significant variability among the ligands and receptors involved. Use of the SELEX technique to evolve nuclease-resistant RNA aptamers for receptor identification is briefly discussed. PMID- 18512342 TI - Calcium regulation and signaling in apicomplexan parasites. AB - Apicomplexan parasites rely on calcium-mediated signaling for a variety of vital functions including protein secretion, motility, cell invasion, and differentiation. These functions are controlled by a variety of specialized systems for uptake and release of calcium, which acts as a second messenger, and on the functions of calcium-dependent proteins. Defining these systems in parasites has been complicated by their evolutionary distance from model organisms and practical concerns in working with small, and somewhat fastidious cells. Comparative genomic analyses of Toxoplasma gondii, Plasmodium spp. and Cryptosporidium spp. reveal several interesting adaptations for calcium-related processes in parasites. Apicomplexans contain several P-type Ca2+ ATPases including an ER-type reuptake mechanism (SERCA), which is the proposed target of artemisinin. All three organisms also contain several genes related to Golgi PMR like calcium transporters, and a Ca2+/H+ exchanger, while plasma membrane-type (PMCA) Ca2+ ATPases and voltage-dependent calcium channels are exclusively found in T. gondii. Pharmacological evidence supports the presence of IP3 and ryanodine channels for calcium-mediated release. Collectively these systems regulate calcium homeostasis and release calcium to act as a signal. Downstream responses are controlled by a family of EF-hand containing calcium binding proteins including calmodulin, and an array of centrin and caltractin-like genes. Most surprising, apicomplexans contain a diversity of calcium-dependent protein kinases (CDPK), which are commonly found in plants. Toxoplasma contains more than 20 CDPK or CDPK-like proteases, while Plasmodium and Cryptosporidium have fewer than half this number. Several of these CDPKs have been shown to play vital roles in protein secretion, invasion, and differentiation, indicating that disruption of calcium-regulated pathways may provide a novel means for selective inhibition of parasites. PMID- 18512343 TI - Trypanosoma cruzi: parasite and host cell signaling during the invasion process. AB - Mammalian cell invasion by Trypanosoma cruzi is a complex process in which various parasite and host cell components interact, triggering the activation of signaling cascades and Ca2+ mobilization in both cells. Using metacyclic trypomastigotes (MT) generated in vitro and tissue culture-derived trypomastigotes (TCT), as counterparts of insect-borne and bloodstream parasites, respectively, the mechanisms of host cell invasion by T. cruzi have been partially elucidated. Distinct sets of molecules are engaged by MT and TCT to enter target cells. MT make use of surface glycoproteins with dual Ca2+ signaling activity, in a manner dependent of T. cruzi isolate. In highly infective MT, the binding of gp82 to its receptor triggers a signaling cascade involving protein tyrosine kinase, phospholipase C and production of inositol 1,4,5-triphosphate, whereas in poorly invasive MT, the mucin-like gp35/50 induces the activation of a signaling route in which adenylate cyclase, generation of cAMP and Ca2+ mobilization from acidocalcisomes are implicated. The host cell signaling pathways activated by MT remain to be determined. Differently from MT, the TCT surface molecules that bind to host cells as a prelude to invasion, such as the glycoproteins of gp85 family, appear to be devoid of signaling properties, but they may induce TCT enzymes, such as oligopeptidase B and cruzipain, to generate Ca2+ signaling factors of parasite or host cell origin. Host cell responses mediated by TGF-beta receptor or integrin family member may also be triggered by TCT. A more complete and detailed picture of T. cruzi invasion needs further investigations. PMID- 18512344 TI - Host cell actin remodeling in response to Cryptosporidium. AB - Cryptosporidium exhibits a complex strategy to invade and establish productive infection sites, involving complimentary parasite and host cell processes. While the work regarding host cell actin remodeling has greatly enhanced our understanding of the molecular pathways involved in the parasite induced actin reorganization, the specific function of host cell actin remodeling is still equivocal. We contend that host cell actin polymerization contributes to the development of productive C. parvum infection sites by generating membrane protrusion events, which may assist in the retention of the parasite at the apical surface within the unique extracytoplasmic niche. With our current understanding of the molecular pathways initiating actin remodeling upon C. parvum interactions with host cells, the next logical step is to determine the upstream events resulting in PI3K activation and the specific role of actin remodeling in parasite development, a process that may have implications beyond host-pathogen interactions. PMID- 18512345 TI - Host cell actin remodeling in response to Trypanosoma cruzi: trypomastigote versus amastigote entry. AB - Trypanosoma cruzi is the protozoan parasite that causes Chagas' disease, a highly prevalent vector-borne disease in Latin America. Chagas' disease is a major public health problem in endemic regions with an estimated 18 million people are infected with T. cruzi and another 100 million at risk (http://www.who.int/ctd/chagas/disease.htm). During its life cycle, T. cruzi alternates between triatomine insect vectors and mammalian hosts. While feeding on host's blood, infected triatomines release in their feces highly motile and infective metacyclic trypomastigotes that may initiate infection. Metacyclic trypomastigotes promptly invade host cells (including gastric mucosa) and once free in the cytoplasm, differentiate into amastigotes that replicate by binary fission. Just before disruption of the parasite-laden cell, amastigotes differentiate back into trypomastigotes which are then released into the tissue spaces and access the circulation. Circulating trypomastigotes that disseminate the infection in the mammalian host may be taken up by feeding triatomines and may also transform, extracellularly, into amastigote-like forms. Unlike their intracellular counterparts, these amastigote-like forms, henceforth called amastigotes, are capable of infecting host cells. Studies in which the mechanisms of amastigote invasion of host cells have been compared to metacyclic trypomastigote entry have revealed interesting differences regarding the involvement of the target cell actin microfilament system. PMID- 18512346 TI - Actin/myosin-based gliding motility in apicomplexan parasites. AB - Apicomplexan parasites move and actively enter host cells by substrate-dependent gliding motility, an unusual form of eukaryotic locomotion that differs fundamentally from the motility of prokaryotic and viral pathogens. Recent research has uncovered some of the cellular and molecular mechanisms underlying parasite motility, transmigration, and cell invasion during life cycle progression. The gliding motor machinery is embedded between the plasma membrane and the inner membrane complex, a unique double membrane layer. It consists ofimmobilized unconventional myosins, short actin stubs, and TRAP-family invasins. Assembly of this motor machinery enables force generation between parasite cytoskeletal components and an extracellular substratum. Unique properties of the individual components suggest that the rational design of motility inhibitors may lead to new intervention strategies to combat some of the most devastating human and livestock diseases. PMID- 18512347 TI - Roles of proteases during invasion and egress by Plasmodium and Toxoplasma. AB - Apicomplexan pathogens replicate exclusively within the confines of a host cell. Entry into (invasion) and exit from (egress) these cells requires an array of specialized parasite molecules, many of which have long been considered to have potential as targets of drug or vaccine-based therapies. In this chapter the authors discuss the current state of knowledge regarding the role of parasite proteolytic enzymes in these critical steps in the life cycle of two clinically important apicomplexan genera, Plasmodium and Toxoplasma. At least three distinct proteases of the cysteine mechanistic class have been implicated in egress of the malaria parasite from cells of its vertebrate and insect host. In contrast, the bulk of the evidence indicates a prime role for serine proteases of the subtilisin and rhomboid families in invasion by both parasites. Whereas proteases involved in egress may function predominantly to degrade host cell structures, proteases involved in invasion probably act primarily as maturases and 'sheddases', required to activate and ultimately remove ligands involved in interactions with the host cell. PMID- 18512348 TI - Roles of naturally occurring protease inhibitors in the modulation of host cell signaling and cellular invasion by Trypanosoma cruzi. AB - Trypanosoma cruzi trypomastigotes rely on the structural diversity of the cruzipain family of cysteine proteases to infect and multiply in nonprofessional phagocytic cells. Herein, we will review studies demonstrating that the interplay of cruzipain with peptidase inhibitors modulate infection outcome in a variety of experimental settings. Studies with a panel of T. cruzi strains showed that parasite ability to invade human smooth muscle cells is influenced by the balance between cruzipain and chagasin, a tight binding endogenous inhibitor of papain like cysteine proteases. Analysis of T. cruzi interaction with endothelial cells and cardiomyocytes indicated that parasite-induced activation of bradykinin receptors drive host cell invasion by [Ca2+]I-dependent pathways. Clues about the mechanisms underlying kinin generation in vivo by trypomastigotes came from analysis of the dynamics of edematogenic inflammation. Owing to plasma extravasation, the blood-borne kininogens accumulate in peripheral sites of infection. Upon diffusion in peripheral tissues, kininogens (i.e., type III cystatins) bind to heparan sulphate chains, thus constraining interactions of the cystatin-like inhibitory domains with cruzipain. The cell bound kininogens are then turned into facile substrates for cruzipain, which liberates kinins in peripheral tissues. Subjected to tight-regulation by kinin-degrading metallopeptidases, such as angiotensin converting enzyme, the short-lived kinin peptides play a dual role in the host-parasite balance. Rather than unilaterally stimulating pathogen infectivity via bradykinin receptors, the released kinins potently induce dendritic cell maturation, thus stimulating type 1 immune responses. In conclusion, the studies reviewed herein illustrate how regulation of parasite proteases may affect host-parasite equilibrium in the course of IT cruzi infection. PMID- 18512349 TI - Biogenesis of and activities at the Toxoplasma gondii parasitophorous vacuole membrane. AB - Apicomplexan parasites like Toxoplasma gondii are distinctive in their utilization of para site encoded motor systems to invade cells. Invasion results in the establishment of the parasitophorous vacuole (PV) within the infected cell. Most apicomplexans complete their intracellular tenure within the infected cell in the PV that is demarcated from the host cytoplasm by the parasitophorous vacuole membrane (PVM). In this chapter I focus on the events surrounding the formation of the PVM and selected activities attributed to it. Its central role as the interface between the parasite and its immediate environment, the host cytoplasm, is validated by the diversity of functions attributed to it. While functions in structural organization, nutrient acquisitions and signaling have been defined their molecular bases remain largely unknown. Several recent studies and the decoding of the Toxoplasma genome have set the stage for a rapid expansion in our understanding of the role of the PVM in parasite biology. Toxoplasma gondii, like all apicomplexan parasites are obligate intracellular pathogens. This family of parasites utilize their own actin-myosin based motor systems to gain entry into susceptible cells establishing themselves, in some cases transiently (e.g., Theileria spp) in specialized vacuolar compartment, the parasitophorous vacuole (PV). The T. gondii PV is highly dynamic compartment defining the replication permissive niche for the parasite. The delimiting membrane defining the parasitophorous vacuole, the parasitophorous vacuole membrane or PVM is increasingly being recognized as a specialized "organelle" that in the context of the infected cell is extracorporeal to the parent organism, the parasite. A systematic study of this enigmatic organelle has been severely limited by several issues. Primary among these is the fact that it is formed only in the context of the infected cell thereby limiting the amount of material. Secondly, unlike other cellular organelles that can often be purified by conventional approaches, the PVM, cannot be purified away from host cell organelles (see below). In spite of these significant obstacles considerable progress has been made in recent years toward understanding the biogenesis of the PVM, identification of its protein complement and the characterization of activities within it. These studies demonstrate that the PVM, on its own and by virtue of its interactions with cellular components, plays critical functions in the structural integrity of the vacuole, nutrient acquisition and the manipulation of cellular functions. In addition it appears that the repertoire of activities at the PVM is likely to be plastic reflecting temporal changes associated with the replicative phase of parasite growth. Finally, the PVM likely forms the foundation for the cyst wall as the parasite differentiates in the establishment of latent infection. As the critical border crossing between the parasite and invaded cell the study of the PVM provides a fertile area for new investigation aided by the recent decoding of the Toxoplasma genome (available at wwww.ToxoDB.org) and the application of proteomic analyses to basic questions in parasite biology. PMID- 18512350 TI - The role of host cell lysosomes in Trypanosoma cruzi invasion. AB - The cell-invasive, trypomastigote form of Trypanosoma cruzi exhibits a unique relationship with lysosomes in target host cells. In contrast to many intracellular pathogens that are adept at avoiding contact with lysosomes, T. cruzi requires transient residence within this acidic organelle for productive infection. The low pH environment of lysosomes facilitates parasite egress from the vacuole and delivery into the host cytosol, a critical step in the T. cruzi developmental program. Recent studies also suggest that early lysosome fusion with invading or recently internalized parasites is critical for cellular retention of parasites. To ensure targeting to host cell lysosomes, T. cruzi trypomastigotes exploit two distinct modes of invasion that rapidly converge in the cell. In this chapter, we summarize the recent progress and changing views regarding the role of host cell lysosomes in the T. cruzi infection process where our discussion is limited to invasion of nonprofessional phagocytic cells. PMID- 18512351 TI - Leishmania invasion and phagosome biogenesis. AB - Whereas bacterial pathogens take over the control of their host cell actin cytoskeleton by delivering an array of protein effectors through specialized secretion systems, promastigotes of the protozoan parasite Leishmania donovani rely entirely upon a cell surface glycolipid to achieve this feat. Here, we review recent evidence that L. donovani promastigotes subvert host macrophage actin dynamics during the establishment of infection and we discuss the potential mechanisms involved. PMID- 18512352 TI - Plasmodium sporozoite passage across the sinusoidal cell layer. AB - Malaria sporozoites must cross at least two cell barriers to reach their initial site of replication in the mammalian host. After transmission into the skin by an infected mosquito, they migrate towards small dermal capillaries, traverse the vascular endothelial layer, and rapidly home to the liver. To infect hepatocytes, the parasites must cross the sinusoidal cell layer, composed of specialized highly fenestrated sinusoidal endothelia and Kupffer cells, the resident macrophages of the liver (Fig. 1). The exact route Plasmodium sporozoites take to hepatocytes has been subject of controversial discussions for many years. Recent cell biological, microscopic, and genetic approaches have considerably enhanced our understanding of the initial events leading to the establishment of a malaria infection in the liver. PMID- 18512353 TI - Transepithelial migration by Toxoplasma. AB - A hallmark of T. gondii infections is passage of parasites across restrictive biological barriers--intestine, blood-brain barrier, blood-retina barrier and placenta-during primary infection or reactivation of chronic disease. Traversal of cellular barriers permits rapid dissemination of parasites to gain access to biologically restricted organs. This process involves active parasite motility and tightly regulated interactions between host cell receptors and parasite adhesins that facilitate paracellular transfer. Mounting evidence also suggests that parasites use migrating leukocytes as Trojan horses to disseminate in the organism while avoiding immune attack. Thus, the interaction of Toxoplasma with biological barriers is a determinant factor of human toxoplasmosis. The elucidation of determinants involved in the process of migration may reveal virulence factors and novel therapeutic targets to combat disease. PMID- 18512354 TI - The microsporidian polar tube and its role in invasion. PMID- 18512356 TI - [Epidemiology, prediction and prevention of type 1 diabetes: not one fiction in Belgium]. AB - The achievements of the Belgian Diabetes Registry (BDR) over a period of almost 20 years are described. This registry is a national and multicentric initiative that collects blood samples and information from diabetes patients and their relatives and treats the results in a confidential and protected way. Not only could the size of the health care problem 'diabetes' finally be quantified and monitored in Belgium for the age group 0-39 years, but in addition a platform was created for the early detection of individuals at high risk of the disease and its complications. This is a conditio sine qua non for the preparation of internationally competitive prevention studies and novel beta cell therapies, that could be effectively launched in Belgium with the help of BDR. PMID- 18512355 TI - Intestinal invasion by Entamoeba histolytica. PMID- 18512357 TI - A molecular genetic update of inherited distal motor neuropathies. AB - Inherited peripheral neuropathies belong to the most common neuromuscular disorders and occur worldwide (1/2500). The best known is Charcot Marie Tooth (CMT), an inherited disorder first described in 1886. Most patients have progressive weakness and wasting of foot and hand muscles. Treatment is currently supportive (braces and foot surgery) and a therapy that fundamentally alters the course of these diseases is still lacking. The involvement of a specific subset of neurons is a key hallmark in the disease process. One subgroup, distal hereditary motor neuropathy (distal HMN) is characterized by the selective loss of motor neurons and/or their long axons in the peripheral nervous system. Apart from the absence of sensory abnormalities, distal HMN closely resembles axonal CMT2. A better understanding of the molecular architecture of the peripheral nerve, the functional pathways, the myelination process and the complex interaction between the axon, the myelinating Schwann cells and muscle is crucial to identify targets for therapeutic interventions. Identification of loci, genes and disease-causing mutations is the first step in this understanding and opens new perspectives for molecular genetic diagnosis. Genotype-phenotype correlations guide the selection of specific mutations suitable for functional analysis in cellular and animal models. The knowledge gained from the molecular genetic and biological research will also help to make progress in the study of acquired peripheral neuropathies. Some of these neuropathies are often therapy-resistant, have a profound influence on the quality of life of the patients, and constitute a financial burden for both the individual and the community. PMID- 18512358 TI - [Immune evasion of alphaherpesviruses]. AB - Alphaherpesviruses represent the largest subfamily of the herpesviruses and comprise many different, closely related pathogens of man and animal, including herpes simplex virus (cold sores, genital lesions) and varicella-zoster virus (chickenpox, shingles) in man, pseudorabies virus orAujeszky's disease virus in pigs (neurological and respiratory symptoms, abortion), equine herpesvirus type 1 (neurological and respiratory symptoms, abortion), and bovine herpesvirus type 1 (respiratory symptoms, abortion). Typical for alphaherpesviruses, and for herpesviruses in general, is their ability to persist in a non-replicative, latent state in their host during its entire lifetime. Specific stimuli can lead to reactivation of these viruses from their latent state, which can lead to renewed spread within and between hosts and recurrent symptoms. This recurrent replication and spread implies that herpesviruses have evolved techniques to delay and avoid recognition and elimination by the immune system, so-called immune evasion mechanisms. In the current manuscript, different alphaherpesvirus immune evasion mechanisms will be reviewed that have been discovered and elucidated at our research group based on pseudorabies virus and that interfere with the antiviral activity of virus-specific antibodies. Investigating immune evasion mechanisms leads to novel insights in the interactions between viruses, host cells, and the immunity, but can also lead to novel avenues in the design of strategies to interfere with viral infections. PMID- 18512359 TI - When the patient is a work of art. PMID- 18512360 TI - Competition and postural confrontation in life, sports, and psychoanalytic treatment--illustrated clinically and in relation to Vladimir Nabokov. AB - The importance of the emotions that can be evoked by (face-to-face and face-to back) postural and visual contrapositions in life and in psychoanalysis-and specifically in relation to sports and games-is delineated and illustrated in clinical and literary material. The latter features aspects of the life and works of the writer Vladimir Nabokov. PMID- 18512361 TI - Berlioz's Symphonie Fantastique: enriching psychoanalytic views of creativity. AB - Focusing on the eighteenth-century French composer Hector Berlioz as he is revealed in his masterpiece Symphonie Fantastique and in his Memoirs, this paper investigates a proposed intermediate area between healthy creativity and paralyzing neurosis. Artists like Berlioz use their work primarily to manage what has been variously labeled as Seelenschmerz, narcissistic injury, and painful fluctuation of internal object representations. Their affectively unprocessed experiences are incorporated into their artwork, so that their heightened experience of Seelenschmerz has particular effects on their audience. Our consideration of these dynamics allows Berlioz's Symphonie Fantastique to regain the truly harrowing impact that it evoked before it became assimilated as a tamed icon of Romantic-era expressiveness. PMID- 18512362 TI - The power of visual memory: the earliest remembered drawing of Alberto Giacometti, Snow White in Her Coffin. AB - Since the time of Freud, many psychoanalysts have seen screen memories and earliest memories as reflecting underlying dynamics. I propose that an earliest remembered artwork is a highly condensed construction similar to a screen memory. Alberto Giacometti's earliest remembered drawing, of Snow White in Her Coffin, contains clues to the artist's personality and references to childhood experience. Giacometti's memory of the drawing done in childhood is a striking condensation of significant biographical events and psychodynamic conflicts, as well as a marker of important unconscious fantasies. The artist's postwar sculptural style, utilizing gaunt figures, epitomizes the final transformation of the psychological meaning of his earliest remembered drawing. PMID- 18512363 TI - Psychoanalytic perspectives on music: an intersection on the oral and aural road. AB - This paper is organized around two ideas. The first invites the reader to consider the importance of music in emotional life, suggesting that for some people, music can have profound, deep, and transformative effects, both in loosening defenses and in deepening the psychoanalytic experience. The second idea is that analysis of the formal properties of music have both specific and overdetermined meanings that share elements with psychoanalytic principles. I suggest that, if the verbal analysis of dreams paves a royal road to the unconscious, the formal properties of music provide an aural road to the same destination. Two clinical vignettes and scenes from Verdi's opera Otello are used to illustrate these interrelated ideas. PMID- 18512364 TI - Dali's homage to Rothko: a defense against fusion with the victim. AB - This paper explores the thematic content of a painting by Salvador Dali in homage to artist Mark Rothko, completed after the latter's suicide. The manifest title of the work suggests respect for Rothko, but the latent intent of the painting, the author suggests, is more of a memoriam to Rothko in a complex identification with him. Among other psychological issues, the author elaborates on two factors that may have played a part in Dali's artistic reaction to Rothko's suicide: the death of the artist's infant brother nine months before his own birth, and his lifelong struggle against fusion with the victim (Orgel 1974a, 1974b). PMID- 18512365 TI - Sublimation and das Ding in Mahler's Symphony no. 8. AB - This paper addresses sublimation in Gustav Mahler's Symphony no. 8 through Lacan's (1986, 1992) notion of das Ding, the Thing. The author reads Lacan as using das Ding, a term taken from Freud, as shorthand for archaic experience. Lacan provides a reference point when he states that "the Kleinian doctrine places the mother's body there" (1992, p. 117). Das Ding refers to unmediated contact with the Other, usually mother, in which traces of a primitive gratification mark the loss of immediacy, point to a lost object, and establish the trajectory of desire. Sublimation is an attempt to bring us into contact with das Ding. PMID- 18512366 TI - The name of the helper: a new look at "Rumpelstiltskin". AB - The fairy tale of Rumpelstiltskin is studied in conjunction with related tales to provide a fuller understanding of its meaning than previous studies have done. The content of these tales depicts ambivalence about childhood magic and ambivalence about adult reality, which are uneasily resolved by eliminating magic and dwelling in a disenchanted world. Enchantment is retained formally in the telling of the story. To the extent that transference is enchantment, similar conflicts occur during psychoanalysis: is the patient better off adhering to transference or relinquishing it--or can it be integrated with day-to-day experience? PMID- 18512367 TI - Hable con Ella (Talk to Her) through the lens of gender. AB - In the 2002 film Hable con Ella (Talk to Her), Spanish writer-director Pedro Almodovar plays with the ambiguity of gender, transcending conventional assumptions about "masculinity" and "femininity." Each of the four main characters holds complex, varied, and, in some cases, gender-bending gender identifications. The theme of gender plasticity is a prominent motif in this film. However, underlying the narrative, there is also a perverse subtext that relies on rigidly binary gender stereotypes to define relationships between men and women. Both these views of gender which operate dialectically, create a complex tapestry through which Almodovar explores his characters' problems in attaining intimacy. PMID- 18512368 TI - Slow progress on safe staffing and healthy workplaces. PMID- 18512371 TI - Election year blues. PMID- 18512372 TI - Home-visiting key to successful nursing model. Interview by Anne Manchester. PMID- 18512373 TI - Robust nursing workforce information--chasing an elusive dream? PMID- 18512374 TI - NZNO through the eyes of an Englishman. PMID- 18512375 TI - Exploring the links between the lack of nursing leadership and poor care. PMID- 18512376 TI - The changing role of charge nurses. PMID- 18512377 TI - What does it take to be a nurse educator in New Zealand? PMID- 18512378 TI - Nurses have an important role in managing intermittent claudication. PMID- 18512379 TI - Sharing responsibility for safe staffing. PMID- 18512380 TI - What's wrong with staffing regulations in aged care? PMID- 18512381 TI - [Clarification of the proposals about the adaptation to Spanish of the standardization of terminology in lower urinary tract function: the Spanish language unites Latin America and Spain]. PMID- 18512382 TI - [New Legal Framework for biodemical research in Spain (act 14/2007)]. PMID- 18512383 TI - [Studies on the kidney and the renal circulation, by Josep Trueta i Raspall (1897 1977)]. AB - INTRODUCTION: Josep Trueta i Raspall, born in Barcelona in 1897, was one of the most outstanding european surgeons of his time. In 1936, after the military coup and during the subsequent Spanish civil war, he was in charge of the Surgery Service at the Hospital de la Santa Creu y Sant Pau and begins to performe a closed method to treat open fractures of the limbs, dramatically lowering the number of gangrenes and amputations between troops and civilians. At the end of the war came the exile, and he was invited by the british Health Service, he teach in Oxford and applies his knowledge on war surgery, was named PhD (Hon) and professor of ortopedics, and creates an school of clinical investigation that generates important findings in bone growing and pathology and, amont other areas, in the knowledge of renal function and kidney injuries induced by shock and blood hypertension. One of the most outstanding fruits of this investigation is the book "Studies on renal circulation", first published in English in 1947 (Oxford), then in Spanish in 1949 (Barcelona). MATERIAL AND METHOD: Study of Josep Trueta's biography and works on kidney function, especially the book "Studies on the renal circulation", relating them to the state-of- the- art of nephrologic studies of his time. RESULTS: Trueta and his group were pioneers in the study of the causes of postraumatic oliguria in war wounds. On the basis of the first studies on crush syndrome, published in 1941, were the kidney failure that followed the extended injuries of the limbs was pointed out, they could demonstrated the changes in renal circulation that followed different injuries to the limbs, such as ischemia, haemorrhage and infections. "Studies on the renal circulation" and other papers published in journals of high scientific impact, shown the renal cortical ischemia with preservation or increase of the medullar and yuxtamedullar circulation that takes place in those pathological situations (Trueta shunt). Although their contributions and hypothesis were not definitives, they served as the basis to studies performed later on, on acute kidney failure secondary to shock, glomerulonephritis, transplant rejection and blood hypertension. COMMENT: Trueta's works are unusual examples of investigations directed to answer clinical questions. What may calls one's attention is the wide field of investigation he moves on, that today would involved several medical specialities, different to his task as orthopedic surgeon. On the other hand, his love by Catalonia, his country, moves him to write "The spirit of Catalonia" (1946), a short text for English-speaking people, on the political and scientific history of Catalonia. Trueta was in the end, an humanistic intellectual with broad interests, reflected on his books on travels or on the life of classic sages such as Luis Vives, Miguel Servet or Cajal. He returned back to Catalonia in 1966 and died in 1977. His book "Studies on the renal circulation", exquisitely illustrated and edited deserves a distinguished place in the European medical and urological historiography. PMID- 18512384 TI - [A preliminar mathematical model for patients with a previous negative prostate biopsy]. AB - INTRODUCTION AND OBJECTIVES: It is usual to identify patients with a negative prostate biopsy who are still at risk of prostate cancer. We try to analyse if the classical variables used in the prostate cancer screening are useful for those patients with a previous negative prostate biopsy, and if there is a possibility for making a nomogram witch would help us in the decision to repeat the biopsy. MATERIAL AND METHODS: We studied 179 patients with at least 1 initial negative biopsy. At each biopsy session we recorded: Patient age, serum prostate specific antigen (PSA), free PSA/total PSA, PSA slope, digital rectal examination, prostate volume, PSA density, cancer suspicion in previous transrectal ultrasounds findings, number of negative cores previously obtained, history of precarcinomatous lesions and time between biopsies. Through Logistic regression analysis we determined the association of each variable a positive biopsy. A nomogram was constructed using all variables and discrimination was calculated as the concordance index. RESULTS: Overall 46% of patients had cancer at the repeated biopsy session. In the univariate analysis: Age, digital rectal examination, prostate volume, PSA density, cancer suspicion in ultrasounds findings, and precarcinomatous lesions were associated with repeat positive biopsy for cancer (all p <0.05). In the multivariate study, age, digital rectal examination, prostate volume and history of precarcinomatous lesions were associated with repeat positive biopsy. A nomogram was constructed that had a concordance index of 0.80. PMID- 18512385 TI - [Is radical cystectomy justified in patients over 75 years old?]. AB - INTRODUCTION AND OBJECTIVES: Radical cystectomy is the standard treatment for invasive bladder cancer. The objectives are to evaluate intraoperative and postoperative complications and to determine overall disease-free interval and overall time to progression in patients over tha age of 75 and to compare these with younger patients. MATERIAL AND METHODS: Between august 1980 and october 2004 , 495 patients underwent radical cistectomy. Patients with palliative surgery were excluded. Patients were divided in two groups according to age: control group (<75 years old) and elderly group (> or =75 years old). RESULTS: Four hundred and two patients were evaluated: 39 patients (35 male and 5 female) in the elderly group and 363 in the control group (321 males and 42 females). Mean age was 76 (range: 75-82) and 62 (range: 35-74) respectively. Mean followup was 38 months in the elderly group and 64 months in the control group. Thirty one patients (80.4%) in the elderly group and 211 patients (58.2%) in the control had non organ-confined tumour in cystectomy specimen (pT3-pT4) (p=0.0096) and ten patients (28.6%) in the elderly and 111 patients (31.4%) in the control group had positive nodes (p=0.84). There were no differences in postoperative surgical complications (p=0.08), postoperative reoperation rate (p=0.58) and postoperative mortality (p=0.28) in both groups. During postoperative time, 11 patients (28%) in the elderly group and 50 patients (13.8%) in the control had medical complications (p=0.03). Fourteen patients (35.9%) in the elderly group and 104 patients (39.4%) in the control group died due to tumour during follow-up (p=0.73). Kaplan-Meier survival curve revealed no differences between two groups in overall disease-free interval and overall time to progression. CONCLUSIONS: Radical cystectomy is a safe and effective treatment in elderly patients with invasive bladder cancer. It is necessary to evaluate co-morbidity in this group because there is an increase in postoperative medical complications. There were no differences between the two groups in overall disease-free interval and overall time to progression. PMID- 18512386 TI - [Orthotopic ileal neobladder. Postoperative management and results]. AB - Urinary diversion after cystectomy have evolved from simple diversion and protection of the upper tracts to functional and anatomic restoration as close as possible to the natural preoperative state. Over the past 15 years, orthotopic reconstruction has evolved from "experimental surgery" to the "preferred method of urinary diversion" in both sexes. Urologist that perform this technique should have an appropriate experience with pelvic surgery and be able to perform a nerve sparing radical cystectomy. Nevertheless, the postoperative management of these patients is more important than the surgical construction if good longterm results are to be achieved. For this reason, a great knowledge about the neobladder's physiology, postoperative complications and their treatment are needed. We review the most important aspects in the postoperative management of patients with ileal neobladder. We also resume the long term outcomes concerning to continence, sexual function, renal impairment, oncologic safety and quality of life. PMID- 18512387 TI - [Renal angiomyolipoma: presentation, treatment and results of 20 cases]. AB - INTRODUCTION: Renal angiomyolipoma is a benign tumour, but its spontaneous rupture may become threatening for patient's live. Both surgery and selective arterial embolization are accepted treatments for this lesion. OBJECTIVES: Analyze renal angyolipoma treated in our center, their clinical outcome and treatment purposed in each case. MATERIAL AND METHODS: We retrospectively analyse 20 cases of patients with renal angiomyolipoma treated in our centre from March 1996 to March 2006. We describe tumour characteristics, treatment followed and results obtained. RESULTS: Patients suffering from tuberous sclerosis (20%) showed multiple bilateral tumours, with size similar to the rest of patients. Diagnosis of tuberous sclerosis was the only factor attached to spontaneous rupture of those lesions. Four of 9 emergency embolized patients (44.4%) required from second treatment because of recurrent haemorrhage, and 4 of the 16 embolization episodes (25%) presented post-embolization syndrome, both with no predicting factors attached. Reduction of less than one third of the inicial diameter was observed in 58.4% of embolized tumours, which used to be multiple, bilateral and of size larger to the rest. No significative differences were observed in plasmatic creatinine after and before treatments. CONCLUSIONS: Renal angiomyolipoma may behave in an aggressive way in patients with tuberous sclerosis. No predicting factors of recurrent haemorrhage or post-embolization syndrome were observed. Both surgery and arterial embolization have proved not to compromise renal function in treated patients. PMID- 18512388 TI - [Preventing urinary fistulas in laparoscopic renal conservative parenchyma surgery with purified bovine serum albumin and glutaraldehyde (bioglue). Initial outcomes]. AB - INTRODUCTION: Urinary fistulas remain an important conservative renal parenchyma surgery (CRPS) complication, especially in central or hiliar tumours and bigger than 4 cm. Herein we present our initial experience preventing fistulae with bioglue (Criolife Inc GA, USA) on laparoscopic CRPS in which urinary tract was opened. PATIENTS AND METHODS: We performed 5 laparoscopic CRPS between September 2005 and February 2006 in which urinary tract was necessarily opened. Previous uretheral catheter, transperitoneal approach, selective arterial control, tumorectomy or heminephrectomy, suturing urinary tract and renal parenchyma and bioglue administration was performed. RESULTS: Median follow up time was 8.2 months (6-12 months). Median surgery time was 138 minutes (105-180 minutes) with a median ischemia time of 45 minutes (35-60). Uretheral catheter was removed before second post-op day in all cases. Average discharged day was 3.8 (3-5 days). One patient required intraoperative transfusion due to breaking Rummel tourniquet and one arterio- calyceal fistula on tenth day pos-op that required selective embolization remained the worst complication. Neither urinary fistulas nor urinomas were reported. CONCLUSIONS: Uretheral cathetesim, suturing urinary tract and parenchyma in an independent fashion and applying some kind of surgical adhesive such as bioglue seems to reduce the urinary fistulae risk in laparoscopic CRPS. PMID- 18512389 TI - [Prognostic factors in localised and regionally advanced renal cell carcinoma]. AB - In the last 10 years, several factors have been identified to confer a prognostic effect on renal cancer outcome. Pathologic stage, nuclear and histologic grade are the most frecuent studied and the most important at this moment. We evaluated those factors and introduced some others, looking for new parameters that could be useful. 96 cases of non methastatic renal cell cancer were included in our study. We found that as was mentioned by other authors pathologic and Furhman stage are the stronger prognostic factors but the presence of palpable tumor, pain and weight lost had significance too. PMID- 18512390 TI - [Urodynamic study in the female urinary incontinence evaluation, with the new MoniTorr MR system (non-multichannel urodynamic with urethral retro-resistance pressure measure). Experience with 100 patients]. AB - OBJECTIVE: To compare the clinical diagnosis of the urinary incontinence with the results of LPP and URP obtained by non-multichannel urodynamic. To evaluate the tolerance of the patients to this examination. PATIENT AND METHOD: Between August and December 2006, 100 urodynamics were made in patients with clinical diagnosis of urinary incontinence, in the Urogynecology Unit of Clinica Las Condes. The clinical diagnosis was compared with the urodynamic results. A subjective tolerance scale was applied (0 was greater tolerance and 10 was maximum discomfort). RESULTS: In 66 patients with clinical diagnosis of IUS, the urodynamic registered 10 Type 0, 2 (I), 23 (II), 1 (III), 17 (II+III), 1 hyperactive detrusor, 5 (0+ hyperactive detrusor). 3 (II+hyperactive detrusor) and 4 (II+III+hyperactive detrusor). In 15 with Mixed urinary incontinence, the urodynamic showed 6 (Type 0), 2 (II), 2 (II+III), 3 hyperactive detrusor, 1 (II+hyperactive detrusor) and 1 (III+hyperactive detrusor). In 16 patients with urgency incontinence, urodynamic showed 2 normal, 4 (II+III), 7 hyperactive detrusor, 1 (II+hyperactive detrusor) and 2 (II+III+hyperactive detrusor). In 3 patients (two vaginal cuff prolapse and one cistocele, degree IV POP-Q) the urodynamic was indicated in the pre-surgery study considering a great potentially incontinence after correction. Respectively, the patients presented: IUS type III+hyperactive detrusor, IUS type II+III and one normal. All patients expressed 2 and 3 degree tolerance (subjective scale: 1 major tolerance and 10 a minor tolerance). CONCLUSIONS: The MoniTorr MR urodynamic is a complementary examination very useful in the study of the urinary incontinence. It allows planning the solution adapted for each case and has a high degree of tolerance in the patients. PMID- 18512391 TI - [Relation between some techniques of hypospadias repair and complications: Universitary San Vicente de Paul Hospital 1986-2005]. AB - OBJECTIVE: The incidence of hypospadias has been increasing in the last decades. Successful hypospadias repair depends on patient factors and surgical technique, a factor closely related to the surgeon's experience. The aim of this descriptive, retrospective study was to determine the preferred technique and its association with postoperative complications. METHODS: Charts of 207 patients who had had hypospadias repair performed between January 1986 and December 2005, in the San Vicente de Paul University Hospital, were reviewed. RESULTS: Hypospadias repair was performed in patients between 1 and 14 years of age. Duckett technique was performed in 38.7%, Mathiew technique in 30% MAGPI technique in 11.6%, Snodgrass in 6.3% and other techniques in 14% of the patients. Postoperative complications included hemorrhagic complications (1.9%), infectious complications (1.9%), urethral diverticular dilatation (3.9%), residual ventral chordee (5.8%), failure of urethroplasty, (8.7%), strictures (23%), and fistulas (41%). Strictures was the most important risk factor for postoperative fistulas (p>0.05). Patients with hypospadias repair with PDS sutures had a lower incidence of postoperative fistulas (p<0.05). Tissue layer over the suture line, age of patient, and number of days with urethral stent was not related to the presence of postoperative complications. CONCLUSION: The importance of known and accepted protective factors in hypospadias repair, like tissue layer over the suture line, age of patient and more or less days with urethral stent, in terms of avoidance of postoperative complications after hypospadias repair, may have to be re evaluated. PMID- 18512392 TI - [Renal transplantation in patients with a vascular aortobifemoral prosthesis]. AB - INTRODUCTION: Nowadays, it is much more common in end stage renal disease patients with vascular grafts, to be kidney transplant candidates. We expose our experience in five cases. MATERIAL AND METHODS: Of all 1,483 kidneys transplanted in our center, 5 recipients had a previous aortobifemoral bypass (2 due to abdominal aortic aneurysm, and 3 due to vascular occlusive disease). We review the clinical features, outcome and complications in these patients. RESULTS: The vascular surgery was done 6 months to 16 years prior to transplantation. The renal transplant was done in iliac fossa with arterial anastomosis to the vascular graft. Surgical complications were: 1 renal artery thrombosis that was treated with thrombectomy, and 1 stricture at the ureterovesical junction. 2 patients dead at 6 months and 7 years with a functioning allograft and 3 patients live with functional allograft at 7 months, 3 years and 7 years. CONCLUSION: Kidney transplantation may be successful in selected patients with aortobifemoral bypass. PMID- 18512393 TI - [Retroperitoneal fibrosis secondary to gastric neoplasia]. AB - The retroperitoneal fibrosis is a very little frequent pathology that habitually includes the abdominal aorta and ureters. Two thirds of cases the etiology is idiopathic and in a third it is secondary to other causes as drugs, generalized infections or diseases and tumors which are considered to be responsible from 8 to 10%. The most frequent is colorectal adenocarcinoma, being anecdotal the gastric location with six described cases to date. We present a case of a patient with gastric cancer and secondary retroperitoneal fibrosis of difficult diagnosis and handling. PMID- 18512394 TI - [Bilateral ureterohydronephrosis secondary to massive vesical hernia]. AB - Bladder hernia is not a rare pathological condition, with a frequency between 0,3 and 3%. Massive bladder hernia is less frequent an very rarely ureterohydronephrosis with this pathology. We will present a case a renal failure secondary to inguinoscrotal bladder hernia with bilateral obstructive uropathy and an analyzed the clinical presentation, the diagnosis and the treatment for those hernias. PMID- 18512395 TI - [Congenital prepubic sinus: case report]. AB - The prepubic congenital sinus is a rare type of urethral duplication of unknown origin. Different embryological theories and classifications has been proposed. Most cases are diagnosed during childhood. The Retrograde Urethrocystography is very important as it determines the diagnosis and helps choosing the best treatment option. We present a case of a prepubic congenital sinus in a 39 years old male who presented with purulent discharge from an accessory meatus in the base of the penis. PMID- 18512396 TI - [Extragonadal germ cell tumor in mediastinum: case report]. AB - The Germ Cell tumors of extragonadal origin are infrequent cases being described in literature less than 1000 cases. The frequent origin sites are mediastinum, retroperitoneum, sacrococcygeal region, and the pineal gland. There was a primary case of germinoma in mediastinum in a 27 years patient. In the image studies a mediastinal mass was demonstrated, a biopsy was taken arriving at the diagnosis. Normal tumor markers, testicular ultrasonography reported the presence of bilateral microlitiasis. It was treating with polychemotherapy and later residual mass was dried. The evolution was satisfactory with complete answer. PMID- 18512397 TI - [Scrotal haematoma as a sign of renal hematoma after ESWL]. AB - We described the case of a patient who presented a large scrotal black discoloration 15 hours after successful extracorporeal lithotripsy of a proximal ureteral calculus. The patient did not present any other clinical symptoms as a sign of subcapsular haematoma. PMID- 18512398 TI - [Immunohistochemical study of colocystoplastie in a case operated 51 years ago]. PMID- 18512399 TI - [Urinary sepsis, severe hydronefrosis and Burkitt lymphoma]. PMID- 18512400 TI - [Retrocaval ureter]. PMID- 18512401 TI - [Cutaneous metastasis of a transitional-cell bladder carcinoma]. PMID- 18512402 TI - [Ectopic kidney. Intravenous urography images]. PMID- 18512403 TI - Cutting-edge tools facilitate care. PMID- 18512404 TI - Protecting health information is of paramount importance. PMID- 18512405 TI - The five Ws of clinical engagement. PMID- 18512406 TI - From the pages of... The Canadian Nurse. PMID- 18512407 TI - Milk product intake: implications for weight control and type 2 diabetes. PMID- 18512408 TI - What goes online is on the record. PMID- 18512409 TI - A new reality. PMID- 18512410 TI - Integrating a standardized assessment into acute care settings: one LHIN's approach. PMID- 18512411 TI - Mapping a path for Canada in nursing documentation. PMID- 18512412 TI - Transforming home and community care. PMID- 18512413 TI - Moving from texts to portals: NurseONE in the classroom. PMID- 18512415 TI - Challenges and change in telehealth. PMID- 18512414 TI - Lynn Nagle is a tireless champion of health informatics. PMID- 18512416 TI - Personality and culture: a comparison of Francophones and Anglophones in Quebec. AB - The authors compared the personality characteristics of Quebec Francophones (25 women, 25 men) and Anglophones (25 women, 25 men) aged 16-64 years. Each participant completed a French or English version of 3 personality inventories that measured (a) extraversion (2 scales), (b) neuroticism (2 scales), (c) psychoticism, (d) lying, (e) openness to experience, (f) conscientiousness, (g) agreeableness, (h) conservatism, and (i) altruism. Francophones scored higher than did Anglophones on psychoticism and on 1 measure of extraversion, and Anglophones scored higher than did Francophones on conservatism, but the 2 groups did not differ on the other personality traits. Men scored higher than did women on psychoticism, and women scored higher than did men on conscientiousness and agreeableness. Conservatism and altruism increased with age. The authors discuss the results in the context of cross-cultural theory and research on personality. PMID- 18512417 TI - Rewarding seniority: exploring cultural and organizational predictors of seniority allocations. AB - The author investigated how organizations in the United Kingdom, the United States, New Zealand, and Germany use seniority in their decisions about rewarding employees. The author asked employees from several organizations to indicate how often their organizations use seniority when making decisions about pay raises, promotions, and dismissals. Results indicated that organizations with egalitarian cultures, economically successful organizations, and public-sector organizations use seniority more often in such decisions. Furthermore, uncertainty avoidance predicted greater use of seniority. The findings are discussed in light of the potential of seniority to maintain and control conflict inherent in open systems and the aging workforce in Western societies. PMID- 18512418 TI - The impact of relational demographics on perceived managerial trustworthiness: similarity or norms? AB - Perceived trustworthiness is a critical antecedent of interpersonal trust, yet researchers have a limited understanding of how such perceptions are generated. The authors used 2 competing perspectives within the relational demography literature--similarity-attraction and relational norms--to empirically examine the effect of demographic differences. Whereas the similarity-attraction account suggests that subordinates will perceive their managers as more trustworthy when managers and staff are similar in demographic attributes, the relational norms account proposes that subordinates will perceive their managers as more trustworthy when their demographic differences follow normative expectations. Data collected from a field study of 178 manager-subordinate dyads in Hong Kong and Macau support the relational norms account in terms of education and organizational rank. The authors discuss the theoretical and practical implications of the study. PMID- 18512419 TI - The hidden impact of conspiracy theories: perceived and actual influence of theories surrounding the death of Princess Diana. AB - The authors examined the perceived and actual impact of exposure to conspiracy theories surrounding the death of Diana, Princess of Wales, in 1997. One group of undergraduate students rated their agreement and their classmates' perceived agreement with several statements about Diana's death. A second group of students from the same undergraduate population read material containing popular conspiracy theories about Diana's death before rating their own and others' agreement with the same statements and perceived retrospective attitudes (i.e., what they thought their own and others' attitudes were before reading the material). Results revealed that whereas participants in the second group accurately estimated others' attitude changes, they underestimated the extent to which their own attitudes were influenced. PMID- 18512420 TI - Effects of social value orientations on fairness judgments. AB - The authors assessed the impact that social value orientations--prosocial (i.e., concerned about outcomes for both oneself and others) versus proself (i.e., concerned about one's own outcome only)--had on fairness judgments in a non negotiation setting. The results indicated that prosocials generally formed fairness judgments in a manner suggested by equity theory: Given the same input as a comparison other, they saw an equal outcome as fairer than a favorable or unfavorable outcome. The fairness determinations of proselfs, however, tended to follow the tenets of self-interest theory: Given the same input as a comparison other, they saw a favorable outcome as fairer than an unfavorable outcome. Contrary to self-interest theory, proselfs did not find a favorable outcome fairer than an equal outcome. These findings indicate that social value orientations differentially affect the evaluation of outcome information in the formation of fairness judgments. PMID- 18512421 TI - Reduced constituent category application in surprising combinations. AB - In a previous study (R. R. C. Hutter & R. J. Crisp, 2005), the authors suggested that one way in which perceivers resolve the dilemma of shared membership of incongruent category combinations is by generating emergent attributes. However, the authors also found evidence that a corresponding process of reduced constituent facilitation occurs. In the present research, the authors aimed to isolate this phenomenon using a response-time methodology. Participants were exposed to subliminal unsurprising or surprising gender-occupation category combination primes. The authors observed facilitation in overall response times on a subsequent lexical decision task to constituent attributes when the combination primed was unsurprising compared with surprising, but only for female and not for male combinations. The authors suggest that perceivers reduce their application of constituent attributes when processing surprising versus unsurprising combinations. PMID- 18512422 TI - The relative effects of competence and likability on interpersonal attraction. AB - Undergraduate students in Singapore (N = 80) learned about the competence (low vs. high) and likability (low vs. high) of a future interaction partner and indicated their attraction toward that stranger. The effect of likability was two times as large as that of competence. Because of the additive effects of the two manipulated factors on attraction, the authors interpreted the preference for lovable fools over competent jerks as an outcome of a generalized supremacy of likability over competence. PMID- 18512423 TI - Taking medical records to the bank. PMID- 18512424 TI - NHIN show and tell. PMID- 18512425 TI - The state connection. State-level efforts in health information exchange. PMID- 18512426 TI - Storage solution. A plan for paper in the transition to electronicdocument management. PMID- 18512427 TI - Watching the puck. Gauging HIM's trajectory in the e-HIM transition. PMID- 18512428 TI - Testing and monitoring access policy controls. The right controls can boost compliance and efficiency. PMID- 18512429 TI - Canada's Interoperable EHR. Infoway chooses the more challenging path, the greater benefits. PMID- 18512430 TI - Complying with the privacy rule during a disaster--Part 2. An overview of interim management. PMID- 18512431 TI - Building an evidence base for high-quality HIM. PMID- 18512432 TI - ICD-10-CM primer. PMID- 18512433 TI - Deciphering dialysis access maintenance coding. PMID- 18512434 TI - EHR support group. HIM professional cofounds Connecticut's first EHR roundtable. PMID- 18512435 TI - A special grouping of papers on landfill gas modeling and management. PMID- 18512436 TI - Methane oxidation in compost-based landfill cover with vegetation during wet and dry conditions in the tropics. AB - The effect of compost and vegetation on methane (CH4) oxidation was investigated during wet and dry conditions in a tropical region. A laboratory-scale experiment was conducted to examine the performance of nonvegetated and vegetated landfill cover systems in terms of CH4 oxidation efficiency. Two types of landfill cover materials (compost and sandy loam) and two species of tropical grasses (Sporobolus virginicus and Panicum repens) were studied for their effect on the CH4 oxidation reaction. It was found that the use of compost as cover material could maintain a high methane oxidation rate (MOR) of 12 mol CH4/m3 x day over a 250-day period. Leachate application showed a positive effect on promoting methanotrophic activity and increasing MOR. A high MOR of 12 mol CH4/m3 x day was achieved when using compost cover with P. repens during wet and dry seasons when leachate irrigation was practiced. In dry conditions, a lower MOR of 8 mol CH4/m3 x day was observed for 80 days. PMID- 18512437 TI - Feasibility of landfill gas as a liquefied natural gas fuel source for refuse trucks. AB - The purpose of this paper is to develop a methodology to evaluate the feasibility of using landfill gas (LFG) as a liquefied natural gas (LNG) fuel source for heavy-duty refuse trucks operating on landfills. Using LFG as a vehicle fuel can make the landfills more self-sustaining, reduce their dependence on fossil fuels, and reduce emissions and greenhouse gases. Acrion Technologies Inc. in association with Mack Trucks Inc. developed a technology to generate LNG from LFG using the CO2 WASH process. A successful application of this process was performed at the Eco Complex in Burlington County, PA. During this application two LNG refuse trucks were operated for 600 hr each using LNG produced from gases from the landfill. The methodology developed in this paper can evaluate the feasibility of three LFG options: doing nothing, electricity generation, and producing LNG to fuel refuse trucks. The methodology involved the modeling of several components: LFG generation, energy recovery processes, fleet operations, economic feasibility, and decision-making. The economic feasibility considers factors such as capital, maintenance, operational, and fuel costs, emissions and tax benefits, and the sale of products such as surplus LNG and food-grade carbon dioxide (CO2). Texas was used as a case study. The 96 landfills in Texas were prioritized and 17 landfills were identified that showed potential for converting LFG to LNG for use as a refuse truck fuel. The methodology was applied to a pilot landfill in El Paso, TX. The analysis showed that converting LFG to LNG to fuel refuse trucks proved to be the most feasible option and that the methodology can be applied for any landfill that considers this option. PMID- 18512438 TI - Methane balance of a bioreactor landfill in Latin America. AB - This paper presents results from a methane (CH4) gas emission characterization survey conducted at the Loma Los Colorados landfill located 60 km from Santiago, Chile. The landfill receives approximately 1 million metric tons (t) of waste annually, and is equipped with leachate control systems and landfill gas collection systems. The collected leachate is recirculated to enable operation of the landfill as a bioreactor. For this study, conducted between April and July 2000, a total of 232 surface emission measurements were made over the 23-ha surface area of the landfill. The average surface flux rate of CH4 emissions over the landfill surface was 167 g x m(-2) x day(-1), and the total quantity of surface emissions was 13,320 t/yr. These values do not include the contribution made by "hot spots," originating from leachate pools caused by "daylighting" of leachate, that were identified on the landfill surface and had very high CH4 emission rates. Other point sources of CH4 emissions at this landfill include 20 disconnected gas wells that vent directly to the atmosphere. Additionally, there are 13 gas wells connected to an incinerator responsible for destroying 84 t/yr of CH4. The balance also includes CH4 that is being oxidized on the surface of the landfill by meth-anotrophic bacteria. Including all sources, except leachate pool emissions, the emissions were estimated to be 14,584 t/yr CH4. It was estimated that less than 1% of the gas produced by the decomposition of waste was captured by the gas collection system and 38% of CH4 generated was emitted to the atmosphere through the soil cover. PMID- 18512439 TI - Greenhouse gas emission potential of the municipal solid waste disposal sites in Thailand. AB - Open dumping and landfilling are the prevalent solid waste disposal practices in Thailand. Surveys on the disposal sites revealed the presence of 95 landfills and 330 open dumps. Methane emission potential at these sites was estimated by three methods. Results of the Intergovernmental Panel on Climate Change (IPCC) method, Landfill Gas Emission model (LandGEM), and closed flux chamber technique were compared. The methane emission potential of 366 Gg/yr using the IPCC method was higher than the estimations of the LandGEM and closed flux chamber method of 115 Gg/yr and 103 Gg/yr, respectively. An understanding of the methane emission potential initiated the analysis of upgrading the open dumps into landfills, adding landfills to meet the future needs and utilization of landfill gases. Upgrading the open dumps to landfills increased the methane emission rates and their utilization potential. Approximately 20 additional landfills may be required to meet future demands. Landfill gas (LFG) utilization appears to be feasible in the large-scale landfills. PMID- 18512440 TI - Estimating national landfill methane emissions: an application of the 2006 Intergovernmental Panel on Climate Change Waste Model in Panama. AB - This paper estimates national methane emissions from solid waste disposal sites in Panama over the time period 1990-2020 using both the 2006 Intergovernmental Panel on Climate Change (IPCC) Waste Model spreadsheet and the default emissions estimate approach presented in the 1996 IPCC Good Practice Guidelines. The IPCC Waste Model has the ability to calculate emissions from a variety of solid waste disposal site types, taking into account country- or region-specific waste composition and climate information, and can be used with a limited amount of data. Countries with detailed data can also run the model with country-specific values. The paper discusses methane emissions from solid waste disposal; explains the differences between the two methodologies in terms of data needs, assumptions, and results; describes solid waste disposal circumstances in Panama; and presents the results of this analysis. It also demonstrates the Waste Model's ability to incorporate landfill gas recovery data and to make projections. The former default method methane emissions estimates are 25 Gg in 1994, and range from 23.1 Gg in 1990 to a projected 37.5 Gg in 2020. The Waste Model estimates are 26.7 Gg in 1994, ranging from 24.6 Gg in 1990 to 41.6 Gg in 2020. Emissions estimates for Panama produced by the new model were, on average, 8% higher than estimates produced by the former default methodology. The increased estimate can be attributed to the inclusion of all solid waste disposal in Panama (as opposed to only disposal in managed landfills), but the increase was offset somewhat by the different default factors and regional waste values between the 1996 and 2006 IPCC guidelines, and the use of the first-order decay model with a time delay for waste degradation in the IPCC Waste Model. PMID- 18512441 TI - A quantitative approach to the traffic air quality problem: the Traffic Air Quality index. AB - The Traffic Air Quality (TAQ) model is a simple tool to estimate traffic fine particulate emissions on roadways (g/km) and can be used for both real-time analysis and for localized conformity analysis ("hot-spot" analysis for nonattainment areas) as defined by 40 CFR 93.123. This paper is a follow-up to a study published earlier regarding the development of the TAQ model. This paper shows how local air quality levels can be a factor in traffic management in nonattainment areas. Similar to the industrial source quotas measured in tons per year, it is proposed that road segments are to be assigned emission quotas (or TAQ indices) measured in pollutant mass emitted per road length (g/km) above which traffic-measures have to be taken to reduce the fine-particulates emissions on such road links. The TAQ model as well as traffic-rerouting measures along with the Intelligent Transportation System (ITS) protocols can be used to have a real-time control of the traffic conditions along expressways to maintain the fine-particulates emissions below the quota assigned per road link and consequently improving the over all local air quality in nonattainment areas. PMID- 18512442 TI - Effect of gaseous chlorine dioxide on indoor microbial contaminants. AB - Traditional and modern techniques for bioaerosol enumeration were used to evaluate the relative efficiency of gaseous chlorine dioxide (ClO2) in reducing the indoor microbial contamination under field and laboratory conditions. The field study was performed in a highly microbially contaminated house, which had had an undetected roof leak for an extended period of time and exhibited large areas of visible microbial growth. Air concentrations of culturable fungi and bacteria, total fungi determined by microscopic count and polymerase chain reaction (PCR) assays, endotoxin, and (1 --> 3)-beta-D-glucan were determined before and after the house was tented and treated with ClO2. The laboratory study was designed to evaluate the efficiency of ClO2 treatment against known concentrations of spores of Aspergillus versicolor and Stachybotrys chartarum on filter paper (surrogate for surface treatment). These species are commonly found in damp indoor environments and were detected in the field study. Upon analysis of the environmental data from the treated house, it was found that the culturable bacteria and fungi as well as total count of fungi (as determined by microscopic count and PCR) were decreased at least 85% after the ClO2 application. However, microscopic analyses of tape samples collected from surfaces after treatment showed that the fungal structures were still present on surfaces. There was no statistically significant change in airborne endotoxin and (1 --> 3)-beta-D-glucan concentration in the field study. The laboratory study supported these results and showed a nonsignificant increase in the concentration of (1 --> 3)-beta-D-glucan after ClO2 treatment. PMID- 18512443 TI - Emission projections for the U.S. Environmental Protection Agency Section 812 second prospective Clean Air Act cost/benefit analysis. AB - Section 812 of the Clean Air Act Amendments (CAAA) of 1990 requires the U.S. Environmental Protection Agency (EPA) to perform periodic, comprehensive analyses of the total costs and total benefits of programs implemented pursuant to the CAAA. The first prospective analysis was completed in 1999. The second prospective analysis was initiated during 2005. The first step in the second prospective analysis was the development of base and projection year emission estimates that will be used to generate benefit estimates of CAAA programs. This paper describes the analysis, methods, and results of the recently completed emission projections. There are several unique features of this analysis. One is the use of consistent economic assumptions from the Department of Energy's Annual Energy Outlook 2005 (AEO 2005) projections as the basis for estimating 2010 and 2020 emissions for all sectors. Another is the analysis of the different emissions paths for both with and without CAAA scenarios. Other features of this analysis include being the first EPA analysis that uses the 2002 National Emission Inventory files as the basis for making 48-state emission projections, incorporating control factor files from the Regional Planning Organizations (RPOs) that had completed emission projections at the time the analysis was performed, and modeling the emission benefits of the expected adoption of measures to meet the 8-hr ozone National Ambient Air Quality Standards (NAAQS), the Clean Air Visibility Rule, and the PM2.5 NAAQS. This analysis shows that the 1990 CAAA have produced significant reductions in criteria pollutant emissions since 1990 and that these emission reductions are expected to continue through 2020. CAAA provisions have reduced volatile organic compound (VOC) emissions by approximately 7 million t/yr by 2000, and are estimated to produce associated VOC emission reductions of 16.7 million t by 2020. Total oxides of nitrogen (NO(x)) emission reductions attributable to the CAAA are 5, 12, and 17 million t in 2000, 2010, and 2020, respectively. Sulfur dioxide (SO2) emission benefits during the study period are dominated by electricity-generating unit (EGU) SO2 emission reductions. These EGU emission benefits go from 7.5 million t reduced in 2000 to 15 million t reduced in 2020. PMID- 18512444 TI - Determination of the potential for release of mercury from combustion product amended soils: Part 1--Simulations of beneficial use. AB - This paper describes a project that assessed the potential for mercury (Hg) release to air and water from soil amended with combustion products to simulate beneficial use. Combustion products (ash) derived from wood, sewage sludge, subbituminous coal, and a subbituminous coal-petroleum coke mixture were added to soil as agricultural supplements, soil stabilizers, and to develop low permeability surfaces. Hg release was measured from the latter when intact and after it was broken up and mixed into the soil. Air-substrate Hg exchange was measured for all materials six times over 24 hr, providing data that reflected winter, spring, summer, and fall meteorological conditions. Dry deposition of atmospheric Hg and emission of Hg to the atmosphere were both found to be important fluxes. Measured differences in seasonal and diel (24 hr) fluxes demonstrated that to establish an annual estimate of air-substrate flux from these materials data on both of these time steps should be collected. Air substrate exchange was highly correlated with soil and air temperature, as well as incident light. Hg releases to the atmosphere from coal and wood combustion product-amended soils to simulate an agricultural application were similar to that measured for the unamended soil, whereas releases to the air for the sludge amended materials were higher. Hg released to soil solutions during the Synthetic Precipitation Leaching Procedure for ash-amended materials was higher than that released from soil alone. On the basis of estimates of annual releases of Hg to the air from the materials used, emissions from coal and wood ash-amended soil to simulate an agricultural application could simply be re-emission of Hg deposited by wet processes from the atmosphere; however, releases from sludge-amended materials and those generated to simulate soil stabilization and disturbed low permeability pads include Hg indigenous to the material. PMID- 18512445 TI - Seasonal and diurnal variability in airborne mold from an indoor residential environment in northern New York. AB - It is well known that characterization of airborne bioaerosols in indoor environments is a challenge because of inherent irregularity in concentrations, which are influenced by many environmental factors. The primary aim of this study was to quantify the day-to-day variability of airborne fungal levels in a single residential environment over multiple seasons. Indoor air quality practitioners must recognize the inherent variability in airborne bio-aerosol measurements during data analysis of mold investigations. Changes in airborne fungi due to varying season and day is important to recognize when considering health impacts of these contaminants and when establishing effective controls. Using an Andersen N6 impactor, indoor and outdoor bioaerosol samples were collected on malt extract agar plates for 18 weekdays and 19 weekdays in winter and summer, respectively. Interday and intraday variability for the bioaerosols were determined for each sampler. Average fungal concentrations were 26 times higher during the summer months. Day-to-day fungal samples showed a relatively high inconsistency suggesting airborne fungal levels are very episodic and are influenced by several environmental factors. Summer bio-aerosol variability ranged from 7 to 36% and winter variability from 24 to 212%; these should be incorporated into results of indoor mold investigations. The second objective was to observe the relationship between biological and nonbiological particulate matter (PM). No correlation was observed between biological and nonbiological PM. Six side-by-side particulate samplers collected coarse PM (PM10) and fine PM (PM2.5) levels in both seasons. PM2.5 particulate concentrations were found to be statistically higher during summer months. Interday variability observed during this study suggests that indoor air quality practitioners must adjust their exposure assessment strategies to reflect the temporal variability in bioaerosol concentrations. PMID- 18512446 TI - Codependencies of reactive air toxic and criteria pollutants on emission reductions. AB - It is important to understand the effects of emission controls on concentrations of ozone, fine particulate matter (PM2.5), and hazardous air pollutants (HAPs) simultaneously, to evaluate the full range of health, ecosystem, and economic effects. Until recently, the capability to simultaneously evaluate interrelated atmospheric pollutants ("one atmosphere" analysis) was unavailable to air quality managers. In this work, we use an air quality model to examine the potential effect of three emission reductions on concentrations of ozone, PM2.5, and four important HAPs (formaldehyde, acetaldehyde, acrolein, and benzene) over a domain centered on Philadelphia for 12-day episodes in July and January 2001. Although NO(x) controls are predicted to benefit PM2.5 concentrations and sometimes benefit ozone, they have only a small effect on formaldehyde, slightly increase acetaldehyde and acrolein, and have no effect on benzene in the July episode. Concentrations of all pollutants except benzene increase slightly with NO(x) controls in the January simulation. Volatile organic compound controls alone are found to have a small effect on ozone and PM2.5, a less than linear effect on decreasing aldehydes, and an approximately linear effect on acrolein and benzene in summer, but a slightly larger than linear effect on aldehydes and acrolein in winter. These simulations indicate the difficulty in assessing how toxic air pollutants might respond to emission reductions aimed at decreasing criteria pollutants such as ozone and PM2.5. PMID- 18512447 TI - Formation characteristics of aerosol particles from pulverized coal pyrolysis in high-temperature environments. AB - The formation characteristics of aerosol particles from pulverized coal pyrolysis in high temperatures are studied experimentally. By conducting a drop-tube furnace, fuel pyrolysis processes in industrial furnaces are simulated in which three different reaction temperatures of 1000, 1200, and 1400 degrees C are considered. Experimental observations indicate that when the reaction temperature is 1000 degrees C, submicron particles are produced, whereas the particle size is dominated by nanoscale for the temperature of 1400 degrees C. Thermogravimetric analysis of the aerosol particles stemming from the pyrolysis temperature of 1000 degrees C reveals that the thermal behavior of the aerosol is characterized by a three-stage reaction with increasing heating temperature: (1) a volatile-reaction stage, (2) a weak-reaction stage, and (3) a soot-reaction stage. However, with the pyrolysis temperature of 1400 degrees C, the volatile- and weak-reaction stages almost merge together and evolve into a chemical-frozen stage. The submicron particles (i.e., 1000 degrees C) are mainly composed of volatiles, tar, and soot, with the main component of the nanoscale particles (i.e., 1400 degrees C) being soot. The polycyclic aromatic hydrocarbons (PAHs) contained in the aerosols are also analyzed. It is found that the PAH content in generated aerosols decreases dramatically as the pyrolysis temperature increases. PMID- 18512448 TI - Development of ambient air quality population-weighted metrics for use in time series health studies. AB - A robust methodology was developed to compute population-weighted daily measures of ambient air pollution for use in time-series studies of acute health effects. Ambient data, including criteria pollutants and four fine particulate matter (PM) components, from monitors located in the 20-county metropolitan Atlanta area over the time period of 1999-2004 were normalized, spatially resolved using inverse distance-square weighting to Census tracts, denormalized using descriptive spatial models, and population-weighted. Error associated with applying this procedure with fewer than the maximum number of observations was also calculated. In addition to providing more representative measures of ambient air pollution for the health study population than provided by a central monitor alone and dampening effects of measurement error and local source impacts, results were used to evaluate spatial variability and to identify air pollutants for which ambient concentrations are poorly characterized. The decrease in correlation of daily monitor observations with daily population-weighted average values with increasing distance of the monitor from the urban center was much greater for primary pollutants than for secondary pollutants. Of the criteria pollutant gases, sulfur dioxide observations were least representative because of the failure of ambient networks to capture the spatial variability of this pollutant for which concentrations are dominated by point source impacts. Daily fluctuations in PM of particles less than 10 microm in aerodynamic diameter (PM10) mass were less well characterized than PM of particles less than 2.5 microm in aerodynamic diameter (PM2.5) mass because of a smaller number of PM10 monitors with daily observations. Of the PM2.5 components, the carbon fractions were less well spatially characterized than sulfate and nitrate both because of primary emissions of elemental and organic carbon and because of differences in measurement techniques used to assess these carbon fractions. PMID- 18512449 TI - Mammography use may partially mediate disparities in tumor size at diagnosis in women with social security disabilities. AB - OBJECTIVE: This study determined the association between screening mammography and tumor size at diagnosis in older women whose original reason for entitlement to Medicare benefits was disability (SSDI). METHODS: A retrospective study of female Medicare beneficiaries older than 69 years diagnosed with breast cancer using Surveillance Epidemiological End Results (SEER)-Medicare linked database. Multiple linear regression techniques were used to determine the effect of screening mammography use on tumor size. MAIN FINDINGS: The total number of women was 413 with SSDI and 8,989 without. Bivariate analysis showed that significantly fewer women with SSDI used screening mammography (45% vs. 38%, P = 0.0006) during the two years prior to diagnosis. Mean tumor size at diagnosis was 2.91 mm (95%, CI = 1.10, 4.73) larger in the group with SSDI. CONCLUSION: This study found that older women whose original reason for Medicare benefits was disability present with larger tumors at breast cancer diagnosis compared to those who were not. Screening mammography may partially mediate the disparity. PMID- 18512450 TI - Gender differences in clinical features of depressed outpatients: preliminary evidence for subtyping of depression? AB - BACKGROUND: Gender differences in depression are usually associated with prevalence, severity, and sometimes with specific syndromes or subtypes. However, a lack of differentiation exists between these factors. AIMS: To disentangle depression severity and the specific items endorsed by men and women and thus explore the presence of gender-specific subtypes. METHOD: A group of 963 men and women treated for depression in the period 1993-2002 were matched on demographic characteristics. This resulted in a group of 353 men and 453 women (N = 806) on which all subsequent analyses were performed: Five instruments were used: the Hamilton Depression Rating Scale (HAM-D), SCL-90 subscales for depression (DEP), anxiety (ANX) and somatic complaints (SOMC), and the Quality of Life Depression Scale (QLDS). Total scores and individual-item scores were compared for men and women using ANOVA. A cluster analysis was performed on the three SCL-90 subscales. The distribution of gender over the clusters was tested with Pearson Chi-square. RESULTS: No gender differences were found in depression severity on the HAM-D. Women reported more symptoms on the DEP and the SOMC (p < 0.01). No gender differences were found on the QLDS. Of the SOMC items, 58% differentiated between men and women, whereas 31% of the DEP items and 30% of the ANX items detected gender differences. Using cluster analysis, a five-cluster solution was found with good face validity and reliability. Men and women were distributed differently over the five clusters such that women were overrepresented in those clusters in which the SOMC was high, while men were overrepresented in clusters in which SOMC was low (p < 0.01). CONCLUSIONS: It may be useful to delineate syndrome of somatic complaints in the context of depression that is more prevalent among women. PMID- 18512451 TI - Internal and external contributors to maternal mental health and marital adaptation one year after birth: comparisons of mothers of pre-term and full-term twins. AB - This longitudinal study examined the contribution of infants' temperament, mother's attachment style, and perceived grandmother's support following delivery, to the psychological mental health and marital adaptation of first time and non-first time Israeli mothers of pre-term (n = 70) and full-term (n = 78) twins, a year later. We collected data for the current study over 2 years (2003 2004). The findings suggested that the extent of mothers' personal and familial stress and their internal resource of attachment style played a crucial role in their mental health and marital adaptation. The external resource of grandmother's support contributed directly to the mothers' marital adaptation, whereas it contributed to their mental health only when infant's temperament was perceived to be difficult. Interestingly, the association between stress and adaptation was stronger among mothers of full-term twins than mothers of pre-term twins. Theoretical and clinical implications are discussed. PMID- 18512452 TI - Abuse history and premenstrual symptomatology: assessing the mediating role of perceived stress. AB - The present study assessed the interrelationships among abuse history (Abuse), perceived stress (Stress), and premenstrual symptom severity reports (PMSR) among female college students (N = 91, 18-25 years old), and determined if Stress mediated the relationship between Abuse and PMSR. Abuse history was noted by 44% of women in this sample, including sexual (25%), physical (11%), or both sexual and physical (8%) abuse. Importantly, results showed significant positive relationships between Abuse, Stress, and PMSR, suggesting Abuse affects psychological and physical aspects of women's health. Overall, women rated PMSR affect symptoms highest, and abused women rated pain and water retention higher than non-abused women. Stress did not fully mediate the relationship between Abuse and PMSR in this study, but accounted for 24% of the variance between these variables. The health implications of these findings are discussed. PMID- 18512453 TI - "How to say it": women's descriptions of pelvic pain. AB - The present research aimed to compare women's descriptions of chronic pelvic pain, when talking about their pain in narrative mode, with the descriptors used in a common pain assessment tool, the McGill Pain Questionnaire (MPQ). Our intention was to see what we could learn about the relationship between words used in these kinds of assessment tools and meanings of pain experience evident in narratives. This New Zealand-based qualitative study used open-ended interviewing to generate women's experiential narratives of pelvic pain. Forty women of European descent were recruited via a randomly selected national prevalence survey on chronic pelvic pain: 33 had chronic pelvic pain that was not associated with dysmenorrhoea or dyspareunia (CPP); 38 had dysmenorrhoea; 29 had dyspareunia; 24 had all three. The study group was aged between 22 and 51 years. The differences that emerged between the words used by women and those used in the MPQ vocabulary are described. Two main findings emerged: a difference in the relative emphasis placed on sensory descriptors and the absence in women's narratives of affective words used in the MPQ. However, a predominance of an affective dimension of pain was evident in women's narratives, which is described. Given the narrative specificity of the experience of pelvic pain, we conclude that assessment tools using the words and phrases evident in narratives of pain would potentially be more useful, and that such a pain assessment tool would ideally be used in association with narrative techniques incorporated into the clinician's interview with women who present with chronic pelvic pain. PMID- 18512454 TI - Examination of body image issues and willingness to be body scanned. AB - This project examined body image issues using 3D body scanning technology. Twenty years of research on body image indicates that though thinness is culturally desirable, possessing an ideal body does not guarantee that women will be satisfied with their bodies. Furthermore, the disconnect continues to widen between the reality of a heavier American woman and the thinner western feminine ideal necessitating the continued need for body image research. This study included 240 female college students enrolled in a mid-sized university. Phase I included a questionnaire of several different measures of body image satisfaction/dissatisfaction, a description of the body scanner including a 3D image, as well as standard demographic questions and estimates of height and weight. Of the initial 240 participants, 85 women agreed to participate in the body scanning portion (Phase II) of the research project. A majority of the women chose an ideal figure thinner than their current figure. Self-esteem scores were significantly correlated with Eating Attitudes Test (EAT) scores. Neither level of body dissatisfaction nor self-esteem scores were significantly associated with willingness to be body scanned, despite the fact that body scanning involves significant body exposure and that the resulting 3D image is highly realistic. Due to the novelty of body scanning technology and its application to body image research, these findings represent an innovative contribution to the field. PMID- 18512455 TI - An ethicist's commentary on anthropomorphism. PMID- 18512456 TI - Distribution of Streptococcus suis capsular types from 2001 to 2007. PMID- 18512457 TI - A comparison of 2 vaccination programs in feedlot calves at ultra-high risk of developing undifferentiated fever/bovine respiratory disease. AB - The aim of this study was to compare 2 vaccination programs in feedlot calves at ultra-high risk of developing undifferentiated fever (UF)/bovine respiratory disease (BRD). At feedlot arrival, 3882 calves were enrolled in the study and randomly allocated to 2 groups, which were housed by group in 12 pens. At the time of allocation, 1 group (MLV3-BT2) received a multivalent, modified-live viral vaccine containing infectious bovine rhinotracheitis virus (IBRV) and types I and II bovine viral diarrhea virus (BVDV), as well as a Mannheimia haemolytica (MH) and Pasteurella multocida bacterin-toxoid. The other group (MLV4-BT1) received a vaccine containing IIBRV, type I BVDV, bovine respiratory syncytial virus, and parainfluenza-3 virus, as well as a MH bacterin-toxoid. At an average of 69 days post arrival, the groups received their respective viral vaccines. The initial UF treatment, overall chronicity, overall wastage, overall mortality, and BRD mortality rates were significantly (P < 0.05) lower in the MLV3-BT2 group than in the MLV4-BT1 group. Average daily gain and the proportions of yield grade Canada 3 and quality grade E carcasses were significantly (P < 0.05) higher in the MLV3-BT2 group than in the MLV4-BT1 group. No significant (P > or = 0.05) difference in the dry matter intake to gain ratio was detected between the 2 groups. In economic terms, there was a net advantage of $20.86 CDN/animal in the MLV3-BT2 group. This study demonstrates that it is more cost effective to use an MLV3-BT2 vaccination program than a MLV4-BT1 vaccination program in feedlot calves at ultra-high risk of developing UF/BRD. PMID- 18512459 TI - Cecal inversion followed by ileocolic intussusception in a cat. AB - A cat was presented with a history of constipation, tenesmus, and malaise. Ultrasonography revealed an ileocolic intussusception and cecal inversion. Surgical findings included an easily reducible intussusception and an inverted cecum that was moderately difficult to reduce. A typhlectomy and bowel plication was performed. The cat had an uneventful recovery. PMID- 18512458 TI - Microbiological and histopathological findings in cases of fatal bovine respiratory disease of feedlot cattle in Western Canada. AB - The aim of this study was to describe the microbiologic agents and pathologic processes in fatal bovine respiratory disease (BRD) of feedlot cattle and to investigate associations between agents and pathologic processes. Ninety feedlot calves diagnosed at necropsy with BRD and 9 control calves without BRD were examined, using immunohistochemical (IHC) staining and histopathologic studies. Mannheimia haemolytica (MH) (peracute, acute, and subacute cases) and Mycoplasma bovis (MB) (subacute, bronchiolar, and chronic cases) were the most common agents identified in fatal BRD cases. Significant associations (P < 0.10) were detected between microbiologic agents and between agents and pathologic processes. When IHC staining was used, 25/26 (96%) of animals that were positive for bovine viral diarrhea virus (BVDV) were also positive for MH; 12/15 (80 %) of animals that were positive for Histophilus somni (HS) were also positive for MB; and all of the animals that were positive for HS were negative for MH and BVDV. This quantitative pathological study demonstrates that several etiologic agents and pathologic processes are involved in fatal BRD of feedlot cattle. PMID- 18512460 TI - Dynamic right ventricular outflow tract (infundibular) stenosis and pectus excavatum in a dog. AB - This is the first published report of a dog with dynamic right ventricular outflow tract (infundibular) stenosis, right ventricular hypertrophy, and pectus excavatum. A juvenile dog presented with a grade V/VI left base systolic heart murmur, tachycardia, and pectus excavatum. Diagnosis of the aforementioned conditions was based on radiography, electrocardiography, and echocardiography. At 9 1/2 wk of age the heart murmur was no longer audible and the right ventricular stenosis and hypertrophy had dissipated and regressed, respectively. Resolution may be associated with growth of the dog. A good prognosis is foreseen. PMID- 18512461 TI - Increased number of skin lesions as a measure of aggression following the mixing of slaughter boars from western Canada assembled for export. AB - A preliminary observational study was conducted to evaluate the animal welfare impacts of holding and mixing on boars; specifically, the need to tusk trim on arrival at assembly. Cull boars assembled in Manitoba from 3 western Canadian provinces were observed without intervention. Although aggression among boars was common, significant physical injury to boars from handling and other boars was rare. Tusk trimming was widely practised in mature boars prior to transport in the population studied. Length of time assembled, number of boars in a pen, temperature, size of boar, and presence of tusk were not associated with change in the skin score of new boars introduced into a pen. Holding groups of previously unfamiliar boars en route to slaughter did not appear to be a significant risk for increased skin lesions in the population studied. Further research is required into the methods and welfare implications to boars subjected to tusk trimming. PMID- 18512462 TI - Suspected transient pseudohypoaldosteronism in a 10-day-old quarter horse foal. AB - A 10-day-old quarter horse colt was presented for signs of disorientation and inability to nurse. Hydronephrosis/hydroureters, with concomitant pyelonephritis and a severe electrolytes disturbance, were diagnosed. The clinical findings closely resembled those described for a syndrome of transient pseudohypoaldosteronism in human neonates. PMID- 18512463 TI - Cosmetic surgery: customer service or professional misconduct. PMID- 18512464 TI - Diagnostic ophthalmology. Squamous cell carcinoma. PMID- 18512465 TI - Acute oropharyngeal effects of full-course radiation treatment of tumors of the head. PMID- 18512466 TI - Baby boomer medicine: an integrated landscape. AB - The country's burgeoning retirement population expects to live longer and better than its predecessors--and as part of that plan, boomers will demand that hospitals offer integrative therapies along with the best of traditional medicine. PMID- 18512467 TI - Worth more than a pound of cure. AB - The national wellness movement is gaining momentum and fans, particularly in dealing with chronic conditions, but some feel it has a punitive side. PMID- 18512468 TI - Assuming quality leadership. AB - Alegent Health in Omaha, Neb., has made leadership in quality a systemwide priority, starting with its senior team and its board of directors. PMID- 18512469 TI - The essential role of boards in executive succession. AB - Only half of succession planning involves hiring the new CEO. The board must also ensure that a solid "talent pipeline" is in place for producing future senior leaders. PMID- 18512470 TI - The rural development imperative: fundraising to 'own' the hospital. AB - Why rural hospitals' community fund-raising efforts matter so much. PMID- 18512471 TI - How to drive a quality dashboard. Interview by Laurie Larson. AB - Quality: What exactly is a quality dashboard? And why should every hospital board have one? PMID- 18512472 TI - A day in the life of a nursing facility administrator. PMID- 18512473 TI - Fitting wheelchairs to residents. PMID- 18512474 TI - Pressure ulcer plan is working. PMID- 18512475 TI - What are you mad about? PMID- 18512476 TI - Where there is smoke, there is ... PMID- 18512477 TI - South Dakota's rural physician shortage: how might it be addressed? PMID- 18512478 TI - Current diagnosis and treatment of polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is the most common endocrinopathy of reproductive-age women. New diagnostic criteria have been established, and evaluation and treatment guidelines have changed in recent years based on the recognition of PCOS as a metabolic disorder. Current recommendations for evaluation and treatment are discussed. PMID- 18512479 TI - Warfarin dosing and genetic testing. PMID- 18512480 TI - Escape from disability (on advanced directives and end-of-life care). PMID- 18512481 TI - Methicillin-resistant Staphylococcus aureus. PMID- 18512482 TI - Extenuating circumstances: bridge to nowhere--circuitous reflections on dementia and life. PMID- 18512483 TI - Impact of atrial fibrillation on immediate and long-term results of mitral balloon valvuloplasty in 531 consecutive patients. AB - BACKGROUND AND AIM OF THE STUDY: Conflicting evidence exists regarding the impact of atrial fibrillation (AF) on the immediate and long-term outcome of mitral balloon valvuloplasty (MBV). The study aim was to investigate such outcome in 531 consecutive patients. METHODS: The immediate and long-term (up to 18 years) clinical and echocardiographic results of MBV of 71 patients with AF at baseline were prospectively collected and compared with those of 460 patients in normal sinus rhythm (NSR). RESULTS: Typically, patients in AF were older (42 +/- 12 years versus 30 +/- 10 years; p <0.0001) and had higher echocardiographic scores (8.45 +/- 1.14 versus 7.95 +/- 1.0; p = 0.005) than those in NSR. In patients with AF, MBV resulted in inferior immediate and long-term results, as reflected by a smaller immediate mitral valve area (MVA) (1.89 +/- 0.23 versus 2.0 +/- 0.3 cm2; p = 0.005), smaller MVA at follow up (1.49 +/- 0.39 versus 1.6 +/- 0.4 cm2; p = 0.037), and a higher restenosis rate (44% versus 30%; p = 0.012). Actuarial freedom from restenosis at 10 years was 67 +/- 6% for AF patients versus 77 +/- 2% for NSR patients (p = 0.11); values at 15 years were 34 +/- 8% and 46 +/- 4%, respectively (p = 0.18). An echo score >8 (p < 0.0001) and previous surgery (p = 0.043) were identified as predictors of restenosis. Actuarial survival at 15 years was lower in AF patients (72 +/- 11% versus 96 +/- 1%; p = 0.029). Likewise, event-free survival was lower in AF patients after 10 years (72 +/- 1% versus 89 +/- 1%; p <0.0001) and 15 years (40 +/- 9% versus 55 +/- 4%; p = 0.128). An echocardiography score > 8 (p < 0.0001) and baseline AF (p = 0.03) were identified as predictors of combined events (p < 0.0001) at follow up. CONCLUSION: AF has a negative impact on the immediate and long-term outcome after MBV. In addition, the presence of AF is a marker of clinical and morphological features associated with inferior results after MBV. PMID- 18512484 TI - Effect of semi-rigid or flexible mitral ring annuloplasty on anterior leaflet three-dimensional geometry. AB - BACKGROUND AND AIM OF THE STUDY: A saddle-shaped mitral annulus may optimize anterior leaflet shape and, in theory, reduce leaflet and chordal stress. Although annuloplasty rings alter native annular height and immobilize the posterior mitral leaflet, their effects on anterior leaflet geometry are unknown. METHODS: Four radiopaque markers were placed on the central meridian of the anterior mitral leaflet (AML), and eight on the mitral annulus, of 20 sheep. Six animals were then implanted with a Carpentier-Edwards Physio ring, and six a Medtronic Duran flexible ring. Eight animals served as controls. All animals were then studied with biplane 60 Hz videofluoroscopy at 7-10 days after surgery. The angle Theta was calculated as the angle between each AML leaflet marker and the annular septal-lateral diameter, while AML marker excursion was expressed as the difference between maximum and minimum angle Theta during the cardiac cycle. The intrinsic AML shape was described by three angles, each between three consecutive leaflet markers from the mid-septal annular marker to the leaflet edge (Phi1-3, from annulus to leaflet edge). RESULTS: Hemodynamic parameters differed only in left ventricular pressure, which was higher in control animals. Anterior leaflet excursion during the cardiac cycle for all four leaflet markers did not change with ring annuloplasty. The intrinsic leaflet angles (Phi1-3) were also unaffected by annular fixation, and thus leaflet shape remained unaltered. CONCLUSION: Neither semi-rigid nor flexible annuloplasty rings affected anterior leaflet excursion or the intrinsic geometry of the AML at end-systole or end diastole. These data suggest that, in normal sheep hearts, annuloplasty rings do not alter anterior leaflet shape and hence do not perturb leaflet stress distribution. PMID- 18512485 TI - Restrictive annuloplasty to treat functional mitral regurgitation: optimize the restriction to improve the results? AB - BACKGROUND AND AIM OF THE STUDY: Today, the 'gold standard' treatment of functional mitral regurgitation (MR) is the subject of much discussion. Although restrictive annuloplasty is currently considered the most reproducible technique, the means by which the degree of annular restriction is optimized remains problematic. The study was designed in order to identify whether the degree of restriction of the mitral annulus could influence early and midterm results following the treatment of functional MR using restrictive annuloplasty. METHODS: A total of 32 consecutive patients with functional MR grade > or = 3+ was enrolled, among whom the mean anterior-posterior (AP) mitral annulus diameter was 39 +/- 3 mm. Restrictive mitral annuloplasty (combined with coronary artery bypass grafting) was performed in all patients using a Carpentier-Edwards Classic or Physio ring (size 26 or 28). The degree of AP annular restriction was calculated for each patient, and correlated with early and mid-term residual MR and left ventricular (LV) reverse remodeling (in terms of LV end-diastolic diameter (LVEDD) and LV end-diastolic volume (LVEDV) reduction). All surviving patients were examined at a one-year follow up. RESULTS: The mean AP mitral annulus restriction achieved was 48 +/- 4%. Intraoperatively, transesophageal echocardiography showed no residual MR in any patient. Before discharge from hospital, transthoracic echocardiography confirmed an absence of residual MR and showed significant LV reverse remodeling (LVEDV from 121 +/- 25 ml to 97 +/- 26 ml; LVEDD from 55 +/- 6 mm to 47 +/- 8 mm). A significant correlation (r = 0.57, p < 0.001) was identified between the degree of AP annulus restriction and LVEDV reduction. A cut-off of annular restriction of 40% (based on AP annulus measurement) correlated with a more significant reverse remodeling. The early postoperative data, with no recurrence of significant MR, was confirmed at a one year follow up examination. CONCLUSION: A marked restriction of the AP mitral annulus diameter (> 40% of preoperative) appears to have a favorable influence on early postoperative LV reverse remodeling, and also allows for complete resolution of functional MR. In addition, 'no tolerance' of early residual MR seems to have a favorable influence on mid-term results, leading to a reduction in the one-year recurrence of significant MR. PMID- 18512486 TI - Long-term echocardiography results of mitral valve repair for mitral valve prolapse. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to assess the long-term durability of mitral valve (MV) repair for MV prolapse using echo-Doppler evaluation. METHODS: Between July 1991 and December 2006, MV repair was performed in 603 patients with nonischemic, severe mitral regurgitation (MR). A subset of 517 patients (mean age 56.3 +/- 12.0 years) with MR caused by leaflet prolapse resulting from degeneration of the MV was subsequently investigated. The main techniques used for MV repair included chordal replacement with ePTFE sutures for 274 of 278 patients with anterior leaflet (AL) prolapse, and leaflet resection for 239 patients with posterior leaflet (PL) prolapse. A prosthetic ring or band was used for annuloplasty in 340 patients, and a band of autologous pericardium in 161; no ring or band was used in the remaining 16 cases. Postoperative serial transthoracic echocardiography was performed for all hospital survivors before discharge, and on at least one occasion after discharge in 507 patients. Echocardiographic follow up was available for up to 15 years (mean 4.4 +/- 3.6 years). Residual MR flow detected by color Doppler echocardiography was classified according to the maximum regurgitant jet area (MRA). RESULTS: The 30 day mortality was 0.57% (three deaths). There were 21 late deaths and 22 reoperations (five of which were re-repair for hemolysis). Kaplan-Meier survival and freedom from reoperation at 14 years were 79.0 +/- 6.0% and 74.5 +/- 9.6%, respectively. Estimates of freedom from severe MR (MRA > or = 7.0 cm2) at five, 10 and 14 years were 94.2 +/- 1.5%, 82.8 +/- 3.6% and 77.5 +/- 5.5%, respectively. Freedom from severe MR at 14 years for 239 patients with isolated PL prolapse was 98.4 +/- 1.6%. CONCLUSION: Echocardiographic follow up of MV repair for MV prolapse demonstrates good long-term results. In particular, the results of MV repair for isolated PL prolapse were excellent. PMID- 18512487 TI - Functional uncoupling of the mitral annulus and left ventricle with mitral regurgitation and dopamine. AB - BACKGROUND: The mitral annulus and left ventricle are generally thought to be functionally coupled, in the sense that increases in left ventricular (LV) size, as seen in ischemic mitral regurgitation (MR), or decreases in LV size, as seen with inotropic stimulation, are thought to increase or decrease annular dimensions in similar manner. The study aim was to elucidate the functional relationship between the mitral annulus and left ventricle during acute MR and inotrope-induced MR reduction. METHODS: Radiopaque markers were implanted on the left ventricle and mitral annulus of five adult sheep. A suture was placed on the central scallop of the posterior mitral leaflet and exteriorized through the atrial-ventricular groove. Open-chest animals were studied at baseline (CTRL), at seconds after pulling on the suture to create moderate-severe 'pure' MR (PULL), and after titration of dopamine until the MR grade was maximally reduced (PULL+DOPA). This process was repeated two to three times for each animal. RESULTS: The MR grade was increased with PULL (from 0.5 +/- 0.01 to 3.4 +/- 0.4, p < 0.01) and decreased after PULL+DOPA (from 3.4 +/- 0.4 to 1.5 +/- 0.9, p < 0.001). PULL resulted in an increase in mitral annular (MA) area, predominantly by an increase in the muscular mitral annulus. PULL+DOPA caused a decrease in MA area, but the LV volume and dimensions were not altered with either PULL or PULL+DOPA. CONCLUSION: The acute geometric response to 'pure' MR and inotrope induced MR reduction was limited to the mitral annulus. Surprisingly, the LV volume and dimensions did not change with acute MR or with inotrope-induced MR reduction. This suggests that, under these two conditions in an ovine model, the mitral annulus and left ventricle are functionally uncoupled. PMID- 18512488 TI - A 'hemispherical' model of aortic valvar geometry. AB - BACKGROUND AND AIM OF THE STUDY: An improved understanding of aortic valvar anatomy could assist in further developing surgical repair of the valve. The study aim was to evaluate the three-dimensional (3-D) geometry of the aortic valve in normal human hearts. METHODS: In eight human cadaver hearts, the aorta and valve were opened longitudinally through the zone of apposition between the right and left coronary leaflets, and photographed. A leaflet of the valve was photographed individually. Valvar circumference (C) was measured as the distance across the aorta at the base of the leaflets. The radius of the valvar orifice (r) was calculated as C/2pi, and the distance between commissures (the peripheral attachments of the zones of apposition between the leaflets at the sinotubular junction) (D') as C/3. The height (h) of the commissural zone of apposition between the leaflets, and also the length (L) of the free-edge, were measured. A model was developed in which three hemispheres, representing the leaflets supported within the sinuses, intersected a cylinder, representing the aorta, all of equivalent radii. The model was tested using dimensional data and paired t tests. RESULTS: In the model, the hemispheres met at the center of the valvar orifice, and each subtended 120 degrees of aortic circumference. The mean (+/- SD) D' (24.7 +/- 2.4 mm) was similar to L (24.5 +/- 2.1 mm), and h (11.9 +/- 1.0 mm) was similar to r (12.0 +/- 1.6 mm) (all p > 0.68), consistent with the model. A series of equations was developed to describe the 3-D geometry of the hemispheres and cylinder in hemispherical and cylindrical coordinates. The areas of coaptation between the leaflets could be calculated, and the intersections between the hemispheres and the cylinder mathematically defined the attachments of the leaflets. Conceivably, the measurement of L could be used to calculate other geometric parameters necessary for valvar competence. CONCLUSION: The normal human aortic valve may be represented as three hemispheres intersecting a cylinder, all with equivalent radii. This simple approach may better define normal anatomic variability, pathologic abnormalities, and strategies for surgical repair. PMID- 18512489 TI - Differential profile of the OPG/RANKL/RANK-system in degenerative aortic native and bioprosthetic valves. AB - BACKGROUND AND AIM OF THE STUDY: Although degenerative calcific aortic valve stenosis is the most common valvular disease among the elderly, neither the etiology underlying the condition nor degeneration of the bioprostheses is yet fully understood. The study aim was to assess the expression profile of those OPG/RANKL/RANK-system determinants known to act as key regulators of bone metabolism and the immune system in calcific aortic valve stenosis and porcine aortic bioprostheses. METHODS: Valve probes from a total of 69 patients (41 with end-stage aortic stenosis, 11 with mild-to-moderate aortic sclerosis, 17 with degenerative porcine aortic bioprostheses) were explanted either during surgery or at autopsy. The presence and localization of OPG, RANKL, RANK and NF-kappaB were analyzed by immunostaining and morphometry. RESULTS: The majority of stenotic and sclerotic valves exhibited cell-bound signals of OPG, RANKL, RANK and NF-kappaB, while bioprostheses showed only sparse signaling. As key findings, the percentage of cells labeled by OPG, RANK and NF-kappaB was increased in sclerotic valves compared with stenotic valves (each p < 0.001), whereas the frequency of RANKL was higher in stenotic compared to sclerotic valves (p < 0.001). As a consequence, the OPG/RANKL ratio was decreased in stenotic (0.83) compared to sclerotic valves (20.2). CONCLUSION: The differential expression profile of specific members of the OPG/RANKL/RANK axis suggests an involvement of their determinants in native valve calcification, but not in the degeneration of porcine bioprostheses. Thus, these mediators of bone homeostasis may represent new targets for a more specified prevention and/or therapy of native aortic stenosis. PMID- 18512490 TI - Native aortic valve endocarditis caused by Candida sake. AB - The case is presented of a native valve endocarditis caused by Candida sake in a 34-year-old farmer with no history of intravenous drug abuse or pre-existing valvular abnormality. The patient presented with septicemia and multiorgan dysfunction syndrome (MODS). Clinical and diagnostic work-up revealed findings of severe aortic regurgitation and large vegetations on the aortic valve. Preoperatively, the patient was treated for three weeks with amphotericin B; when the MODS had improved, open-heart surgery with valve replacement was performed. Intravenous amphotericin B was continued postoperatively for three more weeks; a subsequent decision was taken to administer lifelong suppressive antifungal therapy. PMID- 18512491 TI - Aortic root replacement with a stentless bioprosthesis in osteogenesis imperfecta. AB - Herein is reported a successful surgical case of aortic root replacement for aortic regurgitation in a patient with osteogenesis imperfecta. A 37-year-old male, who had been diagnosed as osteogenesis imperfecta of type IA, underwent mitral valve repair for mitral regurgitation. Severe aortic regurgitation developed four years after surgery. A stentless bioprosthesis was implanted using the full root technique for fear of progressive dilatation of the sinus of Valsalva or aortic dissection, which is well documented in osteogenesis imperfecta. Aortic root replacement with a stentless bioprosthesis for aortic regurgitation represents an alternative procedure of choice for a patient with osteogenesis imperfecta. PMID- 18512492 TI - Aortic valve replacement: is there an implant size variation across Europe? AB - BACKGROUND AND AIMS OF THE STUDY: Prompted by anecdotal evidence and observations by surgeons, an investigation was undertaken into the potential differences in implanted aortic valve prosthesis sizes, during aortic valve replacement (AVR) procedures, between northern and southern European countries. METHODS: A multi institutional, non-randomized, retrospective analysis was conducted among 2,932 patients who underwent AVR surgery at seven tertiary cardiac surgery centers throughout Europe. Demographic and perioperative variables including valve size and type, body surface area (BSA) and early mortality were collected. Group analysis by patient geographic distribution and by annular diameter of the prosthesis utilized was conducted. Patients with a manufacturer's labeled prosthesis size > or = 21 mm were assigned to the 'large' aortic size subset, while those with a prosthesis size < 21 mm were assigned to the 'small' aortic size subset. Effective orifice area indices were calculated for all patients to assess the geographic distribution of patient-prosthesis mismatch. Univariable and multivariable logistic regression analyses adjusting for possible confounding variables were performed. RESULTS: Prostheses with diameter < 21 mm were implanted at almost twice the rate in southern Europe compared to the north (56.4% versus 26.7%, p < 0.01). The mean valve size was also smaller in southern compared to northern European patients (21.6 +/- 2.1 mm versus 23.4 +/- 2.2 mm, p < 0.01). There were no regional differences in the distribution of either gender or BSA. In the multivariable model, south European patients were seven times more likely to receive a smaller-sized aortic valve (OR = 6.5, 95% CI = 4.82-8.83, p < 0.01), and thus the odds of developing patient-prosthesis mismatch were increased two-fold in southern European patients (OR = 1.9, 95% CI = 1.25-2.80, p = 0.02). However, neither geographic distribution nor valve size were significantly associated with operative mortality. CONCLUSION: The study results demonstrated differences in implanted aortic valve size, between the participating northern and southern European countries. Imbalances in the prevalence of rheumatic heart disease, health resource availability and variations in surgical practice throughout Europe might be possible etiological causes. PMID- 18512493 TI - Valve replacement with bileaflet HP-Biplus mechanical prostheses versus standard prostheses: long-term results. AB - BACKGROUND AND AIM OF THE STUDY: At present, no reports are available regarding the hemodynamic assessment and survival of patients undergoing valve replacement with HP-Biplus prostheses. Hence, the present study was designed to acquire this information. The aim was to assess: (i) any potential hemodynamic differences between the HP-Biplus prosthesis and the 'standard' bileaflet mechanical valves (SJM, ATS, Sorin Bicarbon and CarboMedics); and (ii) the incidence of postoperative complications and long-term mortality. METHODS: The data from 242 patients (139 males, 103 females; mean age 58.4 +/- 14.0 years) who survived mitral or aortic valve surgery with mechanical bileaflet standard prostheses (SJM, ATS, Sorin Bicarbon or CarboMedics), between January 1985 and December 2005, were analyzed retrospectively. Evaluations were conducted consecutively with Doppler echocardiography, and compared with 35 patients (20 males, 15 females; mean age 52.2 +/- 12.8 years) who received an HP-Biplus prosthesis and underwent surgery between January 2000 and December 2005. RESULTS: At seven years after surgery, actuarial survival was 40% for the HP-Biplus prosthesis and 84% for the 'standard' prostheses (p < 0.0001). The HP-Biplus prostheses had a higher rate of reoperations for aortic valves (15.2% versus 1.7%; p < 0.003, OR 10.2), a higher rate of prosthesis dysfunction (62.9% versus 7.8%; p < 0.00001, OR 25), and a higher rate of total events (72% versus 21.8%; p < 0.0001, OR 11). CONCLUSION: The present study was the first to compare long-term results of the HP-Biplus prostheses with those obtained with 'standard' mechanical prostheses. Valve replacement with standard prostheses was shown to carry low morbidity and mortality (21.8%), whereas the HP-Biplus prosthesis showed high morbidity and mortality (92%). The data acquired may be very important when selecting prostheses with better hemodynamic characteristics, and show that the HP-Biplus prosthesis, in the authors' opinion, is not suitable for clinical use. PMID- 18512494 TI - Aortic valve replacement with 19-mm bileaflet prostheses in the elderly: left ventricular mass regression and quality of life. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate left ventricular mass (LVM) regression, survival and quality of life in elderly patients after aortic valve replacement (AVR) with small-sized bileaflet prostheses. METHODS: Between September 1988 and September 2005, a total of 147 patients aged > 70 years underwent AVR with 19-mm bileaflet prostheses for aortic stenosis. In order to evaluate the impact of prosthesis-patient mismatch (PPM) on long-term outcome, survivors were allocated to two groups according to the effective orifice area index (EOAI): group A, with EOAI < 0.85 cm2/m2, and group B with EOAI > or = 0.85 cm2/m2. Hospital survivors were interviewed using the SF-36 questionnaire, and the scores compared with those of age- and gender-matched members of the general Italian population. RESULTS: The mean patient age was 74.5 +/- 3.5 years, body surface area (BSA) 1.68 +/- 0.15 m2, and EOAI 0.73 +/- 0.2 cm2/m2. Hospital mortality was 8.8% (n = 13). Actuarial survival was 87.1 +/- 0.028% at one year, 81.3 +/- 0.035% at five years, and 77.2 +/- 0.044% at eight years. Eight-year survival was 74.0 +/- 0.062% in group A and 82.5 +/- 0.064% in group B (p = 0.29). Echocardiographic follow up showed a significant regression of LVM. Scores obtained in the SF-36 test were similar in the two groups, and significantly higher than those of the general Italian population matched for age and gender (p < 0.001 in all domains). CONCLUSION: The implantation of 19-mm bileaflet mechanical prostheses in the elderly allowed LVM regression and a good perceived quality of life. PPM did not influence the long-term survival of these patients. PMID- 18512495 TI - In-vitro testing of three totally supra-annular bileaflet mechanical valves: hydrodynamics in the Sheffield pulse duplicator. AB - BACKGROUND AND AIM OF THE STUDY: In-vivo echocardiographic studies are limited by several confounding factors and technical pitfalls, and consequently the hypothetical differential hydraulic behavior between different prosthetic heart valves has not been identified. However, for surgeons it is essential to know the functional and geometric characteristics of the prostheses to be used. Herein, the in-vitro performance of two new supra-annular bileaflet prostheses--the Medtronic Advantage Supra and Sorin Bicarbon Overline--was compared with that of the 21-mm St. Jude Medical (SJM) Regent valve. METHODS: Three high-performance, production-quality prostheses, including the sewing-ring cuffs, were tested in the aortic chamber of a Sheffield pulse duplicator. The sizes of the prostheses which fitted the 21-mm valve holder were: 21 mm Advantage Supra Medtronic; 19 mm SJM Regent, and 18 mm Sorin Bicarbon Overline. The tests were carried out at a fixed pulse rate (70 beats/min), and at increasing cardiac output (CO) of 2, 4, 5, and 7 l/min. Forward-flow pressure drop, total regurgitant volume, closing and leakage volumes, effective orifice area (EOA) and stroke work loss (SWL) were recorded while the valve was operated at each CO. RESULTS: The SJM Regent and Sorin Bicarbon Overline valves each showed, at increasing CO, significantly lower mean and peak gradients. The calculated EOA and SWL were significantly better with the SJM Regent and Sorin Bicarbon Overline prostheses. The Medtronic Advantage Supra valve showed comparable results only while performing at 2 l/min CO. With regards to the regurgitant fraction, lowest values were observed with the Medtronic Advantage Supra valve. CONCLUSION: This hydrodynamic evaluation model allowed a comparison to be made of the efficiency of recently commercialized bileaflet prostheses, among which the older SJM Regent and the newer Sorin Bicarbon Overline valves demonstrated the best performances. PMID- 18512496 TI - Are initial valve operations in octogenarians still high-risk in the current era? AB - BACKGROUND AND AIM OF THE STUDY: The average age of cardiac patients continues to increase. As more octogenarians undergo surgery during the current era, the outcome of valve surgery was investigated to determine the operative risk in these patients. METHODS: Among 350 patients aged > or = 80 years who had initial surgery between 1998 and 2006, a total of 188 (105 females, 83 males) underwent valve surgery. A prospective analysis was conducted of the collected data. RESULTS: The median age of patients was 82 years (IQR: 81-84 years), and over half of them presented with severe symptoms (NYHA class III/IV; n = 96), controlled heart failure (n = 108), hypertension (n = 101) and coronary artery disease (n = 108). Concomitant coronary artery bypass grafting (CABG) was performed in 89 cases (47%). Perioperative hemodynamic support with inotropes was common (47%). Hospital death after isolated aortic valve replacement (AVR) (n = 89) and mitral valve replacement (MVR) (n = 10) occurred in four patients (4.5%, median additive EuroSCORE 9.0%) and one patient (10%, median additive EuroSCORE 9.8%), respectively. Concomitant CABG led to a doubling of the operative mortality which, for AVR, declined from 5.4% to 3.8% during the latter half of the study period. The median length of stay was 24 h (IQR 21-44 h) in the intensive care unit, and 10 days (IQR 7-14 days) postoperatively. The risk factors for operative mortality were urgent/emergent surgery (HR 3.27, 95% CI 1.12-9.58, p = 0.03), preoperative gastrointestinal disease (HR 3.15, 95% CI 1.12 8.9, p = 0.03), left ventricular ejection fraction <0.30 (HR 4.37, 95% CI 1.29 14.82, p = 0.02), and ischemic time (HR 1.04, 95% CI 1.004-1.07, p = 0.02). CONCLUSION: Elective isolated AVR can be performed with modest operative risk in octogenarians with good left ventricular systolic function. Additional procedures impose long ischemic times and increase the operative risk, as does MVR. Strategies to minimize the complexity and extent of surgery should benefit these patients. PMID- 18512497 TI - Individual patient meta-analysis of self-monitoring of an oral anticoagulation protocol. AB - BACKGROUND AND AIM OF THE STUDY: Oral anticoagulation with vitamin K antagonists is effective for the prevention and treatment of thromboembolic events. Recent systematic reviews have shown that self-monitoring improved the quality of oral anticoagulation therapy (OAT), with patients spending more time in the therapeutic range than traditionally monitored patients, and with a concomitant decrease in the incidence of adverse effects. However, methodological and reporting heterogeneity has limited the strength of the reviews' conclusions. Differences were noted in terms of the assessment of outcome measures and the analysis methods used. For instance, not all used an intention-to-treat analysis, which may have over-inflated the results. Interpretation was limited by missing data: for example, it was not possible to combine mean tests in range, mean time in range, or to determine the level of deviant values. Time-to-event data (e.g., death, thromboembolic events) were reported as numbers of events, which prevented adequate analysis. In order to overcome these limitations and allow further investigation of the data, the study aim is to undertake an Individual Patient Data (IPD) meta-analysis. METHODS AND STUDY DESIGN: The IPD analysis will include data from randomized trials that have compared self-monitoring (self-testing or self-management) OAT versus a control group, and that measured adverse events defined as major hemorrhage, thromboembolism, and death. The data to be requested for each trial will include: outcomes, demographic and psychosocial (e.g., quality of life) data. The primary outcomes of interest will be time to major hemorrhage, thromboembolism, and death. The secondary outcomes will be minor hemorrhage, percentage time within range, percentage tests within range, and patient satisfaction. The primary analysis will be by intention to treat, and multilevel models with patients and trials as the two levels, will be explored to investigate treatment effects on various outcomes. Patient-level covariates will be incorporated into the models in an attempt to account for statistical heterogeneity, as well as to investigate interactions with treatment effect. CONCLUSION: Predictive models should lead to the identification of those most likely to benefit from self-monitoring of oral anticoagulation, and potentially also to a targeted and a more cost-effective use of the intervention. PMID- 18512498 TI - Left thoracotomy for aortic and mitral valve surgery in a case of mediastinal displacement due to pneumonectomy. AB - A 68-year-old male patient with low cardiac output was referred for a redo aortic valve replacement (AVR). Four years earlier, he had undergone a left pneumonectomy for a bronchial carcinoma. Due to the severe shift of the mediastinum into the left chest, as well as to adhesions of the right lung to the sternum, a median sternotomy was thought undesirable. AVR was performed through a left anterior thoracotomy through the fourth intercostal space. The following day, the worsening of a pre-existing mitral insufficiency due to leaflet tethering was diagnosed. Mitral valve annuloplasty was then achieved via a left posterior thoracotomy through the sixth intercostal space. Three weeks later, the patient had recovered a normal hemodynamic and spontaneous breathing. A left thoracotomy should be considered as an alternative approach for valve surgery in case of severe heart displacement. PMID- 18512499 TI - FDA requires HIV warning on popular contraceptives. PMID- 18512500 TI - Your state and vision screening: what's the score? PMID- 18512501 TI - School's out for summer: preparing for next year. PMID- 18512502 TI - Identifying students at risk for type 2 diabetes. PMID- 18512503 TI - Refections for School Nurse Day. PMID- 18512504 TI - School nurses' perceived barriers to diabetes knowledge, communication and management in children with type 1 diabetes. AB - Diabetes is a common chronic illness among children, with 13,000 new cases diagnosed each year. School nurses are in a unique position to facilitate the improvement of health for students with diabetes, but they face many challenges in acquiring diabetes knowledge, communicating with parents and healthcare providers, and managing children and adolescents with diabetes. To learn more about these challenges and the potential role of technology in alleviating them, we administered a survey to school nurses in the state of Maryland. Results indicated that lack of access to knowledge and time constraints were the biggest challenges in providing support and care to children with diabetes. The majority of the school nurses agreed that use of an electronic medium will significantly enhance care and improve knowledge and communication between parents, school nurses and providers. PMID- 18512505 TI - Molecular characterization of one-step self-etching adhesives placed on dentin and inert substrate. AB - PURPOSE: To evaluate the phase separation, extent of dentin demineralization, and curing efficiency of one-step self-etching adhesives placed on dentin and on inert substrate. MATERIALS AND METHODS: The adhesives tested were G-Bond (GB), iBond (IB) and Clearfil S3 Bond (SB). Two groups (2 x 3) of dentin and inert (gold) specimens were prepared for each product. The films formed on inert surfaces were subjected to mild (1st group) or strong air drying (2nd group) and examined with near- and mid-FTIR microspectroscopy. On dentin (1st group), the extent of demineralization (% DM) was evaluated before and after adhesive treatment plus rinsing with water and acetone by mid-FTIR microspectroscopy. The films formed after strong air drying (2nd group) and 20 s of light curing were rinsed with acetone, and the degree of cure (% DC) was measured by micro-multiple internal reflection FTIR spectroscopy. ANOVA was used to assess the statistically significant differences in % DM and % DC (alpha = 0.05). RESULTS: In GB and IB, mild air drying produced more water droplets and monomer separation than strong air drying. SB demonstrated the least separation. IB induced the highest % DM. All the adhesives demonstrated the greatest % DC on dentin. On both substrates SB showed the highest % DC values. CONCLUSION: Phase separation including water and/or monomers was found in all the adhesives. The extent of dentin demineralization and degree of cure varied among the products tested. The degree of cure was higher on dentin than on the inert substrate. PMID- 18512506 TI - Resin tags do not contribute to dentin adhesion in self-etching adhesives. AB - PURPOSE: Self-etching adhesives partly remove or dissolve the dentin smear layer, causing incomplete resin tag formation or low resin tag density. The quantitative contribution of properly formed resin tags to dentin adhesion was evaluated. MATERIALS AND METHODS: We assessed how the presence or absence of resin tags affects tubules of human deep-coronal dentin. G-Bond was used to bond Gradia resin composite. To ensure deep tubule penetration, we used a vacuum exsiccator. For eliminating tag formation, dentin tubules were presealed with adhesive and reverse bonded after finishing. Microtensile bond strength (muTBS) was measured on flat specimens and on Class I cavity floors. Thermocyclic loading was used to estimate the influence of resin tags on long-term behavior. Confocal laser scanning microscopy was used to evaluate adhesive interface dimensions. RESULTS: Hybrid layer thickness, tag length, and tag diameter increased under vacuum treatment. Presealing dentin tubules led to a residual tag area of 3.1% with a tag length of 10.8 microm. Under vacuum, 24.7% of the total dentin surface was covered with tags of 87.8 microm. Low C-factor preparations produced superior muTBS (71.8 to 92.7 MPa) compared with high C-factor Class I cavities (47.0 to 67.6 MPa). Thermocyclic fatigue differed from low to high C-factor situations. In Class I cavities, muTBS significantly decreased after thermocycling. On flat specimens, vacuum infiltration led to reduced muTBS after thermocyclic loading. CONCLUSION: Initially, resin tag formation did not influence the muTBS in either type of C-factor preparation. After thermocyclic loading, muTBS decreased with or without resin tags. Adhesive fracture patterns occurred at the hybrid layer/dentin interface. PMID- 18512507 TI - Hybrid cell death induced by exposure to 2-hydroxyethyl methacrylate (HEMA): an ultrastructural and X-ray microanalytical study. AB - PURPOSE: The aim of this study was to evaluate the ultrastructural characteristics and ionic profile of U937 cells after exposure to 2-hydroxyethyl methacrylate (HEMA) to shed light on the cytotoxicity of this dental adhesive and its relation to mechanisms of cell death. MATERIALS AND METHODS: U937 human monoblastic cells were incubated in RPMI 1640 culture medium and exposed to HEMA at LD50. Structural changes after 5, 15, 30, 60, and 120 min were observed with transmission electron microscopy. Ionic content of Na, K, Cl, Mg, P and S was evaluated by quantitative electron probe X-ray microanalysis. RESULTS: Our results in human monoblastic cell line U937 establish that exposure to HEMA at LD50 led to a singular mechanism of cell death characterized by changes in the morphology and ultrastructure of the cells (cell size, blebs, and organelle structure) compatible with apoptosis, but without changes in nuclear ultrastructure. These findings were consistent with our microanalytical data, which revealed a significant increase in intracellular Na and a decrease in K, along with a significant initial decrease in Cl concentration followed later (120 min) by an increase. CONCLUSION: All three lines of evidence (cell morphology, ultrastructural changes, and ionic profile) showed that HEMA at LD50 led to a hybrid process of cell death. We suggest that apoptosis and necrosis are part of a continuum comprising a single process of cell death. PMID- 18512508 TI - Influence of restorative technique, beveling, and aging on composite bonding to sectioned incisal edges. AB - PURPOSE: The aim of this study was to evaluate the effect of material technique, bevel placement, and aging on the fracture resistance of composite restorations bonded to sectioned incisal edges. MATERIALS AND METHODS: For the retention test, the incisal thirds of 80 mandibular human incisors were sectioned. Ten sound incisors were used as a control group. Teeth were divided into two groups according to storage time (24 h and 180 days with 1000 thermal cycles). In each group, subgroups were randomly formed as follows: beveled or nonbeveled direct resin composite restorations (Adper Single Bond/Filtek Z250) and beveled or nonbeveled indirect composite restorations (prepolymerized Filtek Z250 cemented with Adper Single Bond/Rely X ARC). For each experimental group, 20 specimens were prepared (10 tested after 24 h and the remaining after 180 days). The specimens were subjected to shear testing in a universal testing machine with a crosshead speed of 0.5 mm/min. Failure patterns were analyzed by stereomicroscopy (30X). Data were statistically analyzed with ANOVA and Tukey's and Student's t tests for retention resistance, and with Fisher's exact test for fracture patterns at the 0.05 level of significance for all tests. RESULTS: After 24 h, beveled restorations exhibited higher fracture strength values than nonbeveled restorations and showed resistance similar to the sound teeth. After 6 months, beveled restorations still presented better results than nonbeveled restorations. Thermal cycling and water storage decreased the fracture resistance in the majority of the groups. Adhesive failures were mainly observed in nonbeveled restorations and mixed failures in beveled restorations. CONCLUSION: Within the limitations of the study, it was concluded that storage with thermal cycling decreased fracture resistance, beveling improved fracture resistance, and indirect restorations had a fracture resistance similar to direct restorations. PMID- 18512509 TI - Effects of de- and remineralization of dentin on bond strengths yielded by one-, three-, and four-step adhesives. AB - PURPOSE: To assess the effect of different peri- and intertubular dentin mineralization conditions and etching on shear bond strength in vitro. MATERIALS AND METHODS: One hundred fifty crowns of extracted bovine incisors were embedded in resin and ground to expose the buccal coronal dentin. Sixty specimens were subjected to a demineralizing solution (DS) and another 60 teeth to a bacterial based laboratory caries model (S. mutans, SM). Thirty specimens of each demineralization protocol (DS and SM) were randomly selected and remineralized ( R). Thirty sound dentin specimens served as control (C). Resin composite buildups (Tetric) were bonded after application of one of the following adhesives: a one step self-etching adhesive (Xeno III), and a self-etching adhesive (Syntac Classic) without (three-step) and with prior additional 35% phosphoric acid etching (etch-and-rinse, four-step). Teeth were subjected to shear bond strength testing in a universal testing device at a crosshead speed of 0.5 mm/min (Ultradent method). RESULTS: Bond strength value for group C ranged from 6.3 to 8.4 MPa (p > 0.05). DS and DS-R samples showed in creased bond strength with the one-step adhesive (11.6 MPa, p < 0.05), whereas the three-step adhesive with additional etching showed decreased bond strength (3.2 MPa, p < 0.05). SM samples showed the lowest bond strength of all adhesive systems (range 1.1 to 1.5 MPa, p > 0.05). Remineralization showed no effect on the latter group. CONCLUSION: The degree of mineralization of the dentin is important for adhesion. Additional etching with phosphori acid reduced bond strength of a three-step adhesive. PMID- 18512510 TI - Effect of adhesive system type and tooth region on the bond strength to dentin. AB - PURPOSE: This study evaluated the bond strength of two etch-and-rinse adhesive systems (two- and three-step) and a self-etching system to coronal and root canal dentin. MATERIALS AND METHODS: The root canals of 30 human incisors and canines were instrumented and prepared with burs. The posts used for luting were duplicated with dual resin cement (Duo-link) inside Aestheti Plus #2 molds. Thus, three groups were formed (n = 10) according to the adhesive system employed: All Bond 2 (TE3) + resin cement post (rcp) + Duo-link (Dl); One-Step Plus (TE2) + rcp + Dl; Tyrian/One-Step Plus (SE) + rcp + Dl. Afterwards, 8 transverse sections (1.5 mm) were cut from 4 mm above the CEJ up to 4 mm short of the root canal apex, comprising coronal and root canal dentin. The sections were submitted to push-out testing in a universal testing machine EMIC (1 mm/min). Bond strength data were analyzed with two-way repeated measures ANOVA and Tukey's test (p < 0.05). RESULTS: The relationship between the adhesives was not the same in the different regions (p < 0.05). Comparison of the means achieved with the adhesives in each region (Tukey; p < 0.05) revealed that TE3 (mean +/- standard deviation: 5.22 +/- 1.70) was higher than TE2 (2.60 +/- 1.74) and SE (1.68 +/- 1.85). CONCLUSION: Under the experimental conditions, better bonding to dentin was achieved using the three-step etch-and-rinse system, especially in the coronal region. Therefore, the traditional etch-and-rinse three-step adhesive system seems to be the best choice for teeth needing adhesive endodontic restorations. PMID- 18512511 TI - Sodium hypochlorite as dentin pretreatment for etch-and-rinse single-bottle and two-step self-etching adhesives: atomic force microscope and tensile bond strength evaluation. AB - PURPOSE: The aim of this study was to evaluate the effect of using 5.25% commercial sodium hypochlorite treatment prior to the application of etch-and rinse and self-etching adhesives on dentin surface microtopography and tensile bond strength. MATERIALS AND METHODS: Thirty-two noncarious, nonrestored human third molars were collected. The occlusal enamel of all teeth was removed using diamond disks to expose flat dentin surfaces. The exposed dentin surfaces were abraded using 600-grit SiC disks, to create a uniform dentin smear layer. For AFM characterization, 12 teeth were equally divided into 4 groups according to the proposed dentin surface treatment. Three dentin disks, 2 mm thick, were evaluated per group using tapping mode assessment. Twenty teeth were used for TBS and SEM evaluation and were equally divided into 4 groups, according to the proposed dentin surface treatment. For TBS, 8 dentin/composite slabs, 2 mm thick, were used in each group, while for SEM evaluation 2 slabs were used. Each slab was tested in tension at a crosshead speed of 0.5 mm/min until failure occurred. The samples were examined using SEM operated at 30 kv to evaluate the hybrid layer photographically at 1500X. Statistical analysis was carried out using StatsDirect 2.5.7. One-way ANOVA followed by Tukey-Kramer multiple-comparison post-hoc tests were performed to test the difference between roughness parameters and TBS between groups. RESULTS: Sodium hypochlorite followed by the application of 37% phosphoric acid significantly increased the arithmetic average of the absolute values of surface height deviations (Sa), the surface area ratio which expresses the ratio between the surface area (taking the z height into account) and the area of the flat x,y plane (Sdr), and the surface bearing index (Sbi) parameters, while the application of sodium hypochlorite prior to the application of the self etching primer significantly increased the valley fluid retention index (Svi) parameter. Self-etching primer without sodium hypochlorite pretreatment significantly increased the core fluid retention index (Sci) parameter. Sodium hypochlorite/AdheSE (7.42 +/- 2.16 MPa) significantly increased TBS value compared to other groups. However, no statistically significant difference was found between sodium hypochlorite/Excite (4.68 +/- 1.26 MPa), AdheSE (4.42 +/- 1.36), and Excite (4.06 +/- 1.35). Remnants of smear layer were detected with areas devoid of resin tags in SEM images of samples bonded with AdheSE self etching adhesive, in contrast to samples bonded with sodium hypochlorite followed by AdheSE self-etching adhesive. CONCLUSION: The application of 5.25% commercial sodium hypochlorite with rubbing action for 60 s (total application time 120 s) seems to positively influence the TBS of the self-etching adhesive; however, it has no significant effect on TBS of etch-and-rinse single-bottle adhesive to dentin. The addition of functional roughness parameters to study the dentin surface was shown to be of importance in evaluating the relationship between bond strength and surface topography of conditioned dentin. PMID- 18512512 TI - Dependence of in vitro fracture strength of adhesive core buildup and crown complexes on preparation design and cementation technique. AB - PURPOSE: To evaluate the fracture resistance of teeth restored using an adhesive core material placed under artificial crowns without pins or posts and to assess the effect of different preparation designs and cementation techniques (glass ionomer compared with adhesive cementation) used for the crowns. MATERIALS AND METHODS: Eighty-three intact molars were collected. Sixty-seven teeth were decoronated (test groups and one control group), a 2-mm circular ferrule design was prepared, and four different preparation designs (2-mm- or 1-mm-deep cavities and 2-mm- or 1-mm-thick walls) were used. Three control groups were also established. Cores were built up using an adhesive material. After preparation, standardized artificial crowns (cobalt-chromium alloy) were fabricated. Half of the crowns in the test group (n = 32) were cemented using Panavia (P group); glass-ionomer cement was used for the other half (KC group). All teeth were exposed to 10,000 thermal cycles and loaded until fracture. Statistical analysis was performed, including nonparametric tests (Mann-Whitney U-test) and ANOVA. RESULTS: In the P group, the fracture strength was significantly (p = 0.004) higher (591.75 +/- 177.95 N) than in the KC group (430.18 +/- 193.67 N). The effect of the preparation design was more pronounced in the KC group. In all groups simulating the most moderate type of tooth decay (2-mm-deep cavity and a 2 mm-thick wall), the fracture strength was comparable with that of intact teeth. Results from ANOVA showed that the type of cementation of the crowns affected fracture strength. CONCLUSION: Fracture strengths of adhesive core/crown complexes are greater when an adequate cavity for retention (at least 2 mm deep) is prepared and the crown is luted. PMID- 18512513 TI - SEM analysis of sealant penetration in posterior approximal enamel carious lesions in vivo. AB - PURPOSE: To study the microstructure of sealant penetration in the enamel of in vivo sealed approximal noncavitated incipient caries lesions with and without a preceding bonding step. MATERIALS AND METHODS: A total of 26 proximal noncavitated caries lesions were sealed in vivo, using a sealant in 13 premolars with orthodontic indication of extraction. Each tooth was randomly assigned to mesial or distal surface application of a sealant in the lesion area and in surrounding sound enamel, with or without a bonding system. Four groups were analyzed: a nonbonding group in the lesion area (NBL); a nonbonding group in sound enamel (NBS); a bonding group in the lesion area (BL) and a bonding group in sound enamel (BS). The premolars were extracted after two weeks. All sealed areas were cut and demineralized with 37% hydrochloric acid for 24 h. For each group, the resin tags were observed and measured by scanning electron microscopy. RESULTS: The lesion areas showed a very irregular resin network with twisted and curved tags in contrast to the sound enamel where a regular etching pattern was observed. The length of resin tags in microns for each group (mean +/-SD) was: NBG-L = 4.19 +/- 1.59; NBG-S = 5.49 +/- 2.49; BG-L = 4.57 +/- 1.99; and BG-S = 4.21 +/- 1.87. The differences between the groups were not statistically significant (p = 0.34). CONCLUSION: The use of a bonding system prior to the application of a pit and fissure sealant on both lesion and sound enamel areas does not increase the resin penetration length under non-contaminated conditions. PMID- 18512514 TI - [Oncology in dentistry. Introduction]. PMID- 18512515 TI - [Cancer and oncogenesis]. AB - The term cancer is generally used for malignant neoplasms. A malignant tumour displays invasive growth, i.e. growth beyond its anatomical boundaries, and can result in the formation of distant metastases. Cancer arises from unregulated cell growth, due to alterations in the genome of cells. The cells consequently no longer react to regulatory signals. The accumulation of various genetic defects in a cell ultimately lead to cancer. Cancer is often preceded by a histologically recognizable premalignant stage, called dysplasia. During the past decades, considerable advances have been achieved in characterizing the genetic changes in cells which lead to the beginning of the cell growth. The most important genes involved in these processes are oncogenes, tumour suppressor genes and the so called DNA mismatch repair genes. PMID- 18512516 TI - [Epidemiology, aetiology, and clinical aspects of oral cancer and premalignant lesions]. AB - Since oral cancer can be considered to be a relatively rare disease, dental practitioners will only rarely be confronted with it. Nevertheless, dental practitioners have a key role to play in the early diagnosis and referral to an oral and maxillofacial surgeon. The major part of oral cancers are squamous cell carcinomas, recognizable during routine oral inspection. Early referral is essential since treatment of small, not yet metastasized cancers have the best chance of long-term disease-free survival. Moreover, oral carcinoma is often preceded by white and/or red pre-malignant lesions of the salivary glands. These can similarly be detected by routine inspection. Here too, dentists play an important role in recognizing such diseases and referring patients to an oral and maxillofacial surgeon. PMID- 18512517 TI - [Treatment and prognosis of oral cancer]. AB - Squamous cell carcinoma is the most common variety of malignant oral tumour. Most commonly oral carcinomas occur at the lateral tongue surfaces and at the anterior part of the floor of the mouth. If oral cancer is suspected, a dentist will refer the patient to an oral and maxillofacial surgeon who will perform a biopsy. When the diagnosis squamous cell carcinoma is established, the patient will be referred to a multidisciplinary head and neck oncological centre for additional diagnostics and treatment. Depending upon size, location and extent of the tumour and the presence or absence of regional metastases, the management may include surgical excision, radiotherapy or a combination of surgery and radiotherapy. The prognosis is mainly determined by the size of the tumour and regional lymph node involvement. Therefore, early detection is of utmost importance. PMID- 18512518 TI - [Medical treatment of solid tumours]. AB - The medical treatment of solid tumours depends on many different factors. The choice of drug is stipulated by the tumour type, the stage of the disease and a number of patient characteristics, such as biological age, co-morbidity, and general performance status. The treatment can be curative, palliative or (neo )adjuvant in nature. The groups of drugs which are used are hormones, cytostatics, immune-modulating drugs and (a new group) targeted-drugs consisting of small-molecules and monoclonal antibodies. Only a few tumour types are curable with chemotherapy in an advanced stage. In some tumour types an increase in life expectancy can be achieved; other tumours are hardly or not at all sensitive to medical treatment. Treatment is limited by the side-effects of the drugs. With supporting medication some of the side-effects can be alleviated. With palliative therapy the aim is to improve the general condition by temporarily inhibiting the tumour with minimal side effects. Adjuvant chemotherapy raises the chance of cure after primary treatment with surgery or radiotherapy. PMID- 18512519 TI - [Foci of infection and oral supportive care in cancer patients]. AB - Radiation therapy in the head and neck area and treatment with high dose chemotherapy entail damage to healthy tissue in the mouth. In order to reduce to a minimum the chances of these side effects of cancer treatment developing, it is necessary to carry out oral foci tests prior to oncological therapy. In addition supplementary oral and dental care measures seem to be important in order to limit the side effects of oncological therapy on the teeth, salivary glands and jaw as much as possible. This supportive oral care is not only necessary during, but also for years after the oncology treatment. Therefore not only dental professionals affiliated to oncology teams will have to take care of cancer patients, but also family dentists and dental hygienists. PMID- 18512520 TI - [Reconstruction after surgical treatment of head and neck cancer: surgical and prosthetic possibilities]. AB - Soft and hard tissue defects in the head and neck region after benign or malignant tumour resection, can be reconstructed by surgical techniques, such as tissue transplantation, and/or prostheses. The aim of reconstruction is to restore the original esthetics and functions of the bone and soft tissues that have been resected. The introduction of free vascularized osteomyocutaneous fibula and iliac crest flaps improved the surgical possibilities of reconstructing the mandible and the maxilla. With respect to oral rehabilitation, a reconstruction of the mandible and the maxilla should be carried out in such a way that it provides an adequate base for inserting endosseous implants, which will retain a removable or fixed prosthesis This requires good interdisciplinary planning, in which the plan for prosthetic treatment determines, in part, the choice of reconstruction method. PMID- 18512521 TI - [Dental procedures for patients using oral anticoagulation: new insights]. AB - What treatment is appropriate for a patient who has to undergo an invasive dental procedure if that patient is using medication that influences the blood coagulation system? Should the medication be stopped before the invasive procedure, with the risk of complications involving re-thrombosis? Or should the procedure be carried out without adjusting the medication? What is the risk of bleeding complications? Recent studies appear to indicate that temporarily stopping medication is in many cases unnecessary and can even harm the patient. In recent decades dentists have received a great number of diverse recommendations. In this article recent research in this field is summarized and evaluated. The authors advocate the development of evidence-based clinical guidelines. PMID- 18512522 TI - [Tick-borne disease or not? Do not let yourself be fooled]. AB - Confusion among physicians and patients is increasing regarding the diagnosis and treatment ofLyme borreliosis due to the enormous amount ofambiguous information available and media attention. Some of the dilemmas that physicians encounter are illustrated by 3 patients with a range of symptoms, one of whom was convinced she had Lyme borreliosis. However none of these patients had significant evidence that suggested Lyme borreliosis. Physicians should follow the guidelines developed by the Dutch Institute for Health Care Improvement (CBO) or the Infectious Diseases Society of America rather than 'alternative' guidelines, which are not evidence-based. PMID- 18512523 TI - [Radiofrequency ablation: a worthy addition to the treatment arsenal for liver tumours]. AB - Radiofrequency ablation (RFA) destroys liver tumours by the application of heat. A substantial number of patients with unresectable liver tumours can now be treated by local ablation, with or without partial liver resection. Another great advantage is the possibility of percutaneous RFA treatment, which has minimal morbidity. Although definite proofofits equivalence to other treatments is still lacking, RFA deserves a place in the spectrum of therapies for liver tumours. PMID- 18512524 TI - [Imaging in the diagnosis of colorectal liver metastases and extrahepatic abnormalities]. AB - Imaging using ultrasonography, spiral CT, MRI and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET), plays a major role at two situations during the management of patients with colorectal liver metastases: (a) at the time of the diagnosis and treatment of the primary colorectal tumour, and (b) during the follow-up for the detection of liver metastases and assessing the resectability of these metastases. At the time of the diagnosis and the treatment of the primary tumour, imaging comprising spiral CT or MRI to detect and characterize liver lesions is considered to be the modality of choice. Due to their low prevalence, imaging for the evaluation of lung metastases may be limited to conventional chest radiography. For evaluation of the extrahepatic abnormalities, abdominal and chest CT may be performed in combination with CT of the liver; alternatively a FDG-PET may be performed. During the follow-up of patients treated for colorectal carcinoma, ultrasonography is the most important imaging modality. However, if the liver cannot be adequately imaged by ultrasonography, if there is a raised level ofcarcinoembryonic antigen or irresectability cannot be determined, additional CT or MRI examination will result in more information. PMID- 18512525 TI - [Climate change influences the incidence of arthropod-borne diseases in the Netherlands]. AB - Climate change is associated with changes in the occurrence of arthropod-borne diseases. It is difficult to foresee which arthropod-borne diseases will appear in the Netherlands due to climate change. Climate change influences the prevalence of ticks and may lead to a further increase in Lyme disease and an increased risk of the introduction of rickettsioses. With further warming of the climate there is a real possibility of settlement of the mosquito Aedes albopictus and introduction of the sandfly in the Netherlands. Whether this will lead to circulation of micro-organisms transmitted by these vectors (e.g. West Nile virus, Dengue virus, Leishmania) is not clear. Continued vigilance is necessary, even for vector-borne diseases that appear to be less relevant for the Netherlands. PMID- 18512526 TI - [Diagnostic approach to fever of unknown origin]. AB - Nowadays, fever of unknown origin (FUO) is generally defined as a fever higher than 38-3 degrees C lasting for a period of at least three weeks, in which no definitive diagnosis has been made after a number of obligatory tests. A diagnostic algorithm is proposed in which history taking, physical examination and the obligatory tests are the most important steps in the search for potentially diagnostic clues (PDCs). Next, factitious fever and drug fever should be ruled out. Further diagnostic procedures should be guided by the PDCs. If this does not lead to diagnosis or if there are no useful PDCs, further screening, including 18F-fluorodeoxyglucose positron emission tomography, should be performed. In 30 to 50% of the patients with FUO no diagnosis can be reached. If their clinical condition is stable, waiting to see if new PDCs develop is recommended. Most patients in whom no diagnosis can be made, have a good prognosis. Supportive treatment with NSAIDs can be helpful. Only if patients deteriorate, should other therapeutic trials be considered. PMID- 18512527 TI - [Diagnostic image (368). A child with a facial swelling after a dog bite]. AB - A 4-year-old boy suffered multiple facial dog bites. In the scar of one of these, on the left lower eyelid, a granuloma teleangiectaticum developed. Within a few days, spontaneous regression occurred. One year later, the defect had disappeared completely. PMID- 18512528 TI - [New drugs; exenatide and sitagliptin]. AB - Incretin hormones, secreted upon food intake, play an important role in the regulation of blood glucose levels. In type 2 diabetes mellitus, the incretin response is decreased. Substitution of incretin is a novel pharmacological target which restores postprandial glucose homeostasis. Exenatide is a mimetic of the incretin glucagon-like peptide-I (GLP-I). Sitagliptin is an inhibitor of the enzyme dipeptidyl peptidase 4 (DPP-4), which breaks down GLP-I. Both drugs increase the GLP-I concentration, thereby improving insulin secretion from pancreatic p cells, restoring glycaemic control, preventing beta cell destruction, delaying gastric emptying, and reducing food intake. PMID- 18512529 TI - [Radiofrequency ablation of colorectal metastases to the liver: results since the first application in the Netherlands]. AB - OBJECTIVE: To provide an overview of the morbidity, mortality and survival following the introduction of radiofrequency ablation (RFA) of colorectal liver metastases in the Netherlands. DESIGN: Prospective, descriptive study. METHOD: Between June 1999 and December 2003 in eight hospitals in the Netherlands, 87 patients treated by RFA for colorectal liver metastases were included in the study. The outcome measures were morbidity, 30-day mortality and the percentage local recurrence. RESULTS: In 104 RFA procedures, 199 metastases were ablated; 31 procedures were performed percutaneously and 73 by laparotomy. In 29 procedures, RFA was combined with partial liver resection. The overall postoperative morbidity rate was 19% and the RFA-related morbidity was 14%. 1 patient died following right hemihepatectomy and RFA in the remaining parenchyma (mortality: 1%). Median survival following RFA was 25 months, with a median progression-free survival of 13 months. The overall local recurrence rate was 46%. Since January 2004, this percentage has decreased to approximately 6. Diameter and central location of the metastases were independent risk factors for the development of a local recurrence. CONCLUSION: RFA is an alternative treatment for patients who are not eligible for partial liver resection. The high local recurrence rate in this series reflects the limited experience with this technique during its introduction in the Netherlands. In specialised centres the percentage local recurrence is now 5. Treatment by RFA should always be weighed against the option of partial liver resection and possible (neoadjuvant) chemotherapy. RFA should therefore preferably be carried out in a centre with expertise in the field of liver surgery. PMID- 18512530 TI - [Recurrent pleurisy as sole manifestation of familial Mediterranean fever]. AB - Recurrent pleurisy as sole manifestation offamilial Mediterranean fever. An 18 year-old woman of Turkish descent visited our outpatient department with a 12 year history of recurrent self-limiting febrile attacks accompanied by chest pain. At first the symptoms were attributed to recurrent lower airway infections. However, the persistent nature of the attacks combined with her ethnic background and the spontaneous recovery from the short paroxysmal episodes, led to the consideration of familial Mediterranean fever (FMF). After undergoing treatment with colchicine the patient was free of symptoms. Later it became clear that her 28-year-old brother had the same clinical manifestations of FMF. He was also successfully treated with colchicine. The often long interval from disease onset to correct diagnosis reflects the unfamiliarity of physicians with this disease and the frequency with which it is confused with other syndromes. In patients with paroxysmal febrile attacks and chest pain, especially if they originate from the eastern Mediterranean area, FMF should be considered and colchicine be prescribed to relieve symptoms and prevent amyloidosis. PMID- 18512531 TI - [Primary peritonitis combined with streptococcal toxic shock syndrome following an upper respiratory tract infection caused by Streptococcus pyogenes]. AB - A 52-year-old woman with no previous history of major health problems presented with an acute abdomen and symptoms of shock. Three days earlier she had been diagnosed as having acute laryngitis which was treated with steroids. On admission she was suffering from hypotension, renal failure, liver failure and coagulopathy. Emergency laparotomy revealed purulent fluid spread diffusely throughout the abdominal cavity. Streptococcus pyogenes was grown in culture from this fluid, enabling a diagnosis of streptococcal toxic shock syndrome (STSS) with primary peritonitis to be made. This combination is rare, and has been described only a few times. Only one other patient is known in whom this combination was preceded by respiratory symptoms. The treatment consists of abdominal lavage, intravenous administration of antibiotics and immunoglobulins, and support for renal function, liver function, respiration and coagulation. PMID- 18512532 TI - [The placing of an unborn child under formal supervision where there is doubt about the safety of the home situation]. AB - Requests to place an unborn child under formal supervision was made in the course of two pregnancies. The first woman was 27 years old, she had a history of schizophrenia, forensic psychiatric care, and a personality disorder with impulsive aggressive behaviour. The second patient was 36 years old. She had a bipolar disorder due to which her firstborn had been placed in foster care. In the first case, formal supervision for the unborn child ensued. In the second case the request was initially denied, but due to the disordered domestic situation was granted ten days after birth. Prior to birth, a relevant risk assessment based on maternal characteristics can be made. In the Netherlands it is possible to place a foetus under formal supervision after 24 weeks gestation. This may prevent hospitalization of a healthy newborn in an unhealthy environment which is poor in stimuli. It also prevents the stressful situation that may arise when parents threaten to take their newborn child from the hospital, pending the inquiry into the domestic situation. PMID- 18512533 TI - [International medical graduates in Dutch health care: the new assessment procedure]. AB - On December 1, 2005 in the Netherlands, a new procedure was introduced to assess international medical graduates (IMGs) with a diploma acquired outside the European Economic Area (EEA). This procedure includes (a) general tests on the active and passive use of Dutch medical language, English reading proficiency, basic IT skills and knowledge of the Dutch health care system, and (b) a specific set of tests of medical competence, including knowledge of basic sciences, clinical knowledge and clinical skills. IMGs who wish to get their diploma acknowledged and be registered as a physician are required to complete this assessment. With the introduction of this procedure, the Netherlands have joined a minority of countries inside and outside Europe with setting high standards for intake procedures. It is advocated that all European countries should devise such procedures, as a European Directive (2005/36/EC) on the recognition of professional qualifications prohibits the assessment of medical graduates with a diploma that is recognised in another EEA country. PMID- 18512534 TI - [Health care insurance for Africa]. PMID- 18512535 TI - Demanding individually safe drugs today: overcoming the cross-labeling legal hurdle to pharmacogenomics. PMID- 18512536 TI - Essay: the limits of conscience: moral clashes over deeply divisive healthcare procedures. PMID- 18512537 TI - Mandating a human papillomavirus vaccine: an investigation into whether such legislation is constitutional and prudent. PMID- 18512538 TI - Vascular birthmarks. AB - Vascular birthmarks can be classified into hemangioma and vascular malformations. Hemangioma are frequent tumours of early infancy demonstrating endothelial hyperplasia, a history of rapid neonatal growth and slow involution during later childhood. Treatment of hemangioma is dependent of stage and type of the lesion. Given the current availability of drugs, lasers, and other techniques to treat hemangioma safely, philosophy of "benign neglect" should not be considered anymore. Vascular malformations show a normal endothelial turnover, being present at birth and growing commensurately with the child. Exact diagnosis by employing modern diagnostic means,which are able to differentiate low-flow from high flow lesions is important for further therapeutic management. Beside conservative treatment strategies, use of laser, sclerotherapy, interventional embolization and surgical treatment are possible management options. Patients should receive multidisciplinary care in qualified vascular centres. PMID- 18512539 TI - Evidence-based indications for thrombophilia screening. AB - Thrombophilic defects have been shown to be associated with an increased risk of venous thrombosis, fetal loss, and gestational complications. The knowledge about the clinical relevance of thrombophilic defects is increasing, and evidence-based indications for thrombophilia screening are therefore discussed in this review. Selective thrombophilia screening based on previous personal and/or family history of venous thromboembolism is more cost-effective than universal screening in all patient groups evaluated. In the majority of patients with acute venous thrombosis, the results of thrombophilia screening do not influence the duration of oral anticoagulation. The only patient population who clearly profits from thrombophilia screening in this situation are patients with a newly diagnosed antiphospholipid syndrome, because prolonged anticoagulation can avoid the high incidence of recurrence in this patient population. Because of the increased risk of venous thrombosis during pregnancy and the puerperium, thrombophilia screening is indicated in selected patients with a previous history of venous thrombosis or a positive family history. Significant associations with early and late pregnancy loss are observed for carriers of the heterozygous factor V Leiden mutation, the heterozygous prothrombin-mutation G20210A and anticardiolipin antibodies, while protein S deficiency is significantly associated with late pregnancy loss. Antithrombotic drugs like UFH, LMWH or low-dose aspirin may have a potential therapeutic benefit in patients with recurrent pregnancy loss and thrombophilia, but placebo-controlled, multicenter trials are urgently needed to clarify this issue. Although a supra-additive effect for the risk of venous thrombosis is observed between oral contraceptives and some thrombophilias, the absolute incidence of venous thromboembolism is low in premenopausal women and mass screening strategies are therefore unlikely to be effective. While antiphospholipid antibodies are known to be associated with arterial thrombosis, screening for heritable thrombophilias is not useful in arterial thrombosis, although subgroup analysis indicates that they may play a role particularly in young patients and children. PMID- 18512540 TI - Superficial vein thrombophlebitis--serious concern or much ado about little? AB - Superficial vein thrombophlebitis (SVTP) appears in two distinct forms: varicose vein thrombophlebitis (TP) represents the principal cause. It is characterized by a large thrombus in a varicose vein and a modest inflammatory process localized in the vessel surrounding but not in its wall. Rarely, SVTP affects a non varicose vein. Abundant intima proliferation and media fibrosis with non important thrombosis are the hallmark of this form which may be associated with a systemic disease. Although SVTP is perceived as trivial and benign coexistence of (mostly distal) deep venous thrombosis (DVT), propagation to popliteal or femoral DVT, and even pulmonary embolism (PE) have been reported. Data for prevalence vary greatly: 6-53% for coexistence, 2.6-15% for propagation, and 0-33% for (asymptomatic) PE. Risk factors for these complications are those known for DVT. SVTP is diagnosed in a clinical setting but ultrasonography is useful to check for concomitant DVT. Anticoagulant treatment is mandatory if DVT is present and thrombectomy should be considered in cases of thrombus propagation into the deep veins. Historical therapy of uncomplicated SVTP consists of compression with bandages or stockings and local or systemic anti-inflammatory agents. Low molecular-weight heparin (LMWH) has been given in high-prophylactic doses and found equally effective when compared with anti-inflammatory agents and full therapeutic dose LMWH. Prophylactic saphenous vein ligation alone was found less effective than conservative therapy. Ligation combined with stripping proved the potential of eliminating at once all problems associated with SVTP but was associated with a complication rate of 10% or higher. Careful patient selection and saphenous vein thrombectomy prior to stripping may be the clue for better results. PMID- 18512541 TI - Lymphoedema and lipoedema of the extremities. AB - Lymphoedema is a clinical manifestation of an impaired lymphatic drainage with accumulation of lymphatic fluid. Lipoedema is characterized by bilateral enlargement of the legs and/or arms due to abnormal deposition of fatty tissue, which accumulates fluid. Conservative treatment including compressions garments and lymphatic drainage is suitable to prevent ongoing clinical deterioration although both diseases cannot be cured. The ability to properly diagnose lymphoedema and lipoedema is crucial to prevent the significant morbidity and loss of quality of life that is associated with this condition. It is imperative that patients with lymphoedema are referred to specially trained healthcare professionals to ensure optimal treatment. Continuous therapy with strict compliance of the patients is essential, and premature interruption is the most frequent mistake. Lipoedema is a different entity but patients are still fighting for acceptance. The mutual relation of lipoedema and obesity and the poor knowledge of the underlying mechanisms limit the acceptance of lipoedema as a relevant disease. PMID- 18512542 TI - Acute aortic dissection--vascular emergency with numerous pitfalls. AB - Acute aortic syndrome comprises acute aortic dissection, aortic intramural haematoma and penetrating atherosclerotic ulcers of the aortic wall. It ranks, after acute coronary syndrome, as one of the most frequent acute life-threatening differential diagnoses of chest pain. Chances of survival would probably be good in the large majority of cases, assuming optimal therapeutic management including rapid diagnostic evaluation followed by immediate and appropriate treatment is provided. However, actual mortality rate in these patients is still currently higher than 40%, despite medical and surgical progress. This unfavourable prognosis for the most frequent variant of acute aortic syndrome--aortic dissection--is due to the wide variability of clinical symptoms. These are often initially unspecific and frequently lead to delays in establishing the correct diagnosis, possibly first recognised at autopsy. Even after a timely, correct diagnosis, there is still a considerably high mortality rate following surgery, even with younger patients. Whenever acute aortic dissection is suspected a diagnostic imaging study should immediately be obtained. In addition to CT angiography, transesophageal echocardiography is recommended, due to its flexibility, as the diagnostically most useful tool in this context. Based on three case reports of acute aortic dissection this paper critically discusses the problems in making a correct and timely diagnosis and also provides an overview of the current state of knowledge in the areas of pathophysiology, epidemiology, clinical symptomatology as well as appropriate case-related management and prognosis of acute aortic dissection. PMID- 18512543 TI - L-arginine plasma levels and severity of idiopathic pulmonary arterial hypertension. AB - BACKGROUND: Idiopathic pulmonary arterial hypertension (iPAH) is a rare disease of unknown aetiology characterized by a poor prognosis. Impairment of nitric oxide (NO) synthesis or NO-induced vasorelaxation has been suspected to play a role in the development of iPAH. This study was performed to investigate possible correlations between the plasma levels of the NO-related aminoacids L-arginine, L citrulline and N-hydroxy-L-arginine (L-NHA) and the severity of iPAH. METHODS: In twelve iPAH patients hemodynamics were measured by right heart catheterization, and plasma levels of L-arginine, L-citrulline and L-NHA were determined in blood samples from the pulmonary artery, peripheral artery and peripheral vein by high performance liquid chromatography analysis. In eight of twelve patients a six minute walk test was performed. RESULTS: Plasma levels of L-arginine strongly correlated to right atrial pressure, cardiac output, cardiac index, mixed-venous oxygen saturation, six minute walk data and NYHA functional class at all sites of blood sampling (p < 0.05). CONCLUSIONS: The results suggest a possible role of the NO precursor L-arginine in the pathogenesis of iPAH. PMID- 18512544 TI - Surgery of inferior vena cava associated malignant tumor lesions. AB - BACKGROUND: Tumor lesions of the inferior vena cava (IVC) can originate from the vein or can develop by malignant tumor infiltration from the surrounding tissue. In this context, particular attention should be paid to tumor lesions with pegs into or within the IVC. The aim of this series of a single surgical center was to analyze the perioperative management, the individual-specific and -adapted surgical technique, as well as the outcome including prognostic considerations in IVC-associated malignant tumor lesions. PATIENTS AND METHODS: Over a 6-year time period, all consecutive patients with IVC-associated malignant tumor lesions and their patient- and finding-specific characteristics were registered, data and parameters of the diagnostic and therapeutic management were documented, and both the short- and long-term outcomes (complication rate, perioperative morbidity/mortality, tumor recurrence rate, survival) were assessed with periodic follow-up investigations. RESULTS: Overall, 12 patients were enrolled in the study from 1/1/2001-31/12/2006:6 primary IVC-tumors (leiomyosarcomas, 50%) and 6 secondary IVC-tumors (2 retroperitoneal tumor lesions, 16.7%, 3 renal cell carcinomas 25% and 1 carcinoma of the adrenal gland, 8.3%). 4 of the secondary tumors had pegs into the IVC. The RO resection rate was 83%. The perioperative morbidity was 33%; whereas, the hospital mortality was 8.3% (n = 1). Surgical reconstruction of IVC was achieved in each case (100%). There was a mean postoperative observation period of 20 months (range, 1-58 months). Complete follow-up documentation was obtained for all of the patients (100%). Three patients experienced recurrent tumor growth (27.5% out of n = 11). While the overall mortality through the follow up observation period was 27.5%, the tumor specific mortality was 16%. CONCLUSIONS: The primary surgical aim is RO resection to provide a long-term outcome with no tumor recurrence including the reconstruction of the IVC based on a reasonable risk-to-benefit ratio. The favorable outcome of this case series demonstrates that IVC-associated tumor lesions can be approached if there is an appropriate expertise of the surgical team, a sufficient perioperative management and an adequate financial background with a reasonable survival rate. The variable prognosis of the various tumor lesions depends on tumor entity, stage, resection status and individual risk factors. PMID- 18512545 TI - Hemodialysis access surgery--is there an increased risk of acquiring hepatitis C virus compared to other elective vascular interventions? AB - BACKGROUND: To evaluate whether dedicated access surgeons might have a significantly higher risk of acquiring hepatitis C infection compared to other vascular surgeons by assessing the prevalence of hepatitis C patients who are on chronic hemodialysis and to compare the frequency to patients undergoing elective vascular interventions. PATIENTS AND METHODS: A retrospective chart and data analysis of all patients on chronic hemodialysis was conducted. As a comparative group, the prevalence of anti-HCV antibodies and positive HCV RNA PCR among patients admitted for elective vascular surgery was assessed. RESULTS: Of 285 patients on chronic hemodialysis, 202 (71%) were had both tests (antibody test for HCV and specific HCV RNA PCR testing). 5% (n = 11; CI 95 = 3-10%) were antibody positive, and 4% (n = 8; CI 95 = 2-8%) were also PCR positive and therefore infectious. One patient was acutely infected. Of 4963 vascular surgical patients, 1141 (23%) had an anti-HCV antibody ELISA test and specific HCV RNA PCR testing. 0.4% (n = 4; CI 95 = 0.1-1%) were antibody positive and 0.2% (n = 2; CI 95 = 0.03-0.7%) were also PCR positive and hence infectious. No acutely infected patient was detected in this population. The chance of operating on a HCV positive and infectious patient among hemodialysis patients was almost 27 times higher than among elective vascular surgical patients (P < 0.0001; OR = 26.56; CI 95 = 5.42-253.40). CONCLUSIONS: Dedicated hemodialysis access surgeons have a higher risk to acquire hepatitis C infection compared to vascular surgeons performing all other elective vascular surgical interventions. To identify early infected surgeons operating on high risk HCV patient collectives and to start rapid treatment, PCR testing at regular intervals would be advisable. PMID- 18512546 TI - Symptomatic bilateral carotid artery stenoses 7 years after coronary artery bypass surgery in a young patient with familial hypercholesterolemia. AB - We present a patient suffering from homozygous familial hypercholesterolemia, who developed coronary artery disease in the age of 15 treated with three vessels coronary artery bypass grafting (CABG) operation. 7 years after CABG surgery symptomatic bilateral carotid artery stenoses were observed. Both internal carotid arteries were stented. The combination of pharmacologic therapy with life long LDL-apheresis is the current treatment of choice. PMID- 18512547 TI - Duplex ultrasound and efficacy criteria in foam sclerotherapy from the 2nd European Consensus Meeting on Foam Sclerotherapy 2006, Tegernsee, Germany. AB - RATIONALE: The spread of foam sclerotherapy has resulted in the renaissance of sclerotherapy as a non-invasive treatment method for varicosis. An expanded European expert committee meeting in Tegernsee in April 2006 was prompted by new findings and continuous further development of the method and worked especially on the topics "The role of (duplex) ultrasound in Foam sclerotherapy" and "Evaluation of therapeutic effects of foam Sclerotherapy". It was felt that these criteria are"non-specific" to foam sclerotherapy and would possibly also be suitable for other endovenous ablative procedures. The organisers of the 2nd European Consensus Meeting on foam sclerotherapy (2nd ECMFS) were then asked to publish these recommendations in this separate publication. The entire recommendations of the 2nd ECMFS are published in an extensive overview in this journal (VASA 2008; 37; Supplement 71: 1-32). METHODOLOGY: The 29 participants were sent a comprehensive questionnaire in advance covering all the relevant aspects of foam sclerotherapy. The organisers drew up various preliminary statements on the basis of the results. During the meeting itself the participants revised and/or approved and/or rejected these statements. For the "non-specific" topics, two working groups were given the task of conducting the concluding assessment of these items. Their final results were presented in March and April 2007. RESULTS: For foam sclerotherapy, duplex ultrasound is important in pre-treatment diagnosis, treatment monitoring/guidance, post-treatment efficacy evaluation and surveillance. In the pre-treatment diagnosis of varicose veins, the exact localisation of the insufficient saphenous, communicating and perforating veins is important. Duplex ultrasound is the accepted gold standard for this purpose. The application of ultrasound imaging during foam sclerotherapy increases the safety of accessing the vein in certain indications, and it can help when making a decision concerning the foam volumes to be injected, the patients' position or specific movements the patients should perform. Following treatment, the findings of duplex ultrasound, the clinical findings and the patients' symptoms can be arranged according to the recommended definitions. This allows grading of the therapeutic outcome and enables a better comparability between different treatment protocols or different treatments. Besides the evaluation of treatment success, duplex ultrasound is the method of choice to exclude or confirm complications such as deep venous thrombosis or disease progression. PMID- 18512548 TI - [Utilization of national research achievements with more effectiveness]. PMID- 18512549 TI - [A potential mechanism for impaired wound healing--cutaneous environmental disorders in diabetes mellitus]. AB - Impaired wound healing in diabetes is a significant clinical problem which is thought to be associated with neuropathy and angiopathy previously . The present study indicates that accumulation of glucose and glycometabolic products in skin tissue, as the result of glycometabolic disorders, which contributes to cutaneous environmental alterations in diabetes mellitus, and subsequently induces the abnormal cell behaviors, cytokine alteration and matrix modification. Thus, diabetic neuropathy and angiopathy might be regarded as the pathological outcome of cutaneous environmental alterations. In conclusion, glycometabolism disorders could be described as one of the initial events for the alteration involving in the underlying cutaneous disorder which impair healing process. The related research focuses on the initial event of controlling disorders in wound healing and therefore contribute to providing the strategy of treatment as based on these approaches. PMID- 18512550 TI - [Surgical treatment of multiple pressure scores]. AB - OBJECTIVE: To summarize the methods and results of the surgical treatment of patients with multiple pressure sores. METHODS: Twenty-one patients with 56 multiple pressure sores, hospitalized from January 2001 to May 2007, were treated with transfer of various skin flaps together with skin grafting. The pressure sores were respectively located in sacrococcygeal region (21 wounds), ischial tuberosity (14 wounds), greater trochanter of femur (13 wounds) and other sites (8 wounds). All the patients were given systemic supporting treatment in perioperative period and early debridement . The wounds were repaired with flaps, fascio-musculocutaneous flaps, or free skin grafts according to their size, depth, position and the condition of adjacent skin and soft tissue. Continuous irrigation, negative pressure suction, regular posture changes in turning frame after operation were also emphasized. RESULTS: Twenty-five wounds were repaired by fascio-cutaneous flap or myocutaneous flap with healing rate of 90%. Thirteen wounds were repaired by adjacent regional flap with healing rate of 85%. Eight wounds were treated with direct suturing,among which 6 healed completely. Ten wounds were treated with free skin grafting,among whom 7 healed completely. Among 9 delayed healing wounds, 4 wounds healed after debridement and suturing or free skin transplantation for second time, 4 wounds healed by dressing change in a short time, and in the last a chronic sinus remained. Follow-up over 6 months, multiple pressure sores recurred in 3 patients. CONCLUSION: Enhancing systemic supporting treatment in perioperative period, using fascio-cutaneous flap or myocutaneous flap to repair multiple sores, followed by continuous irrigation and negative pressure suction after operation, and regular postural change on turning frame, contribute a rate of success for management of multiple pressure sores. PMID- 18512551 TI - [Effects of advanced glycosylation end products on the biological behavior of neutrophils]. AB - OBJECTIVE: To investigate the effects of advanced glycation end products (AGE) on the biological behavior of neutrophils in vitro, to look for the relationship between accumulation of AGE and abnormal inflammation in wound healing in diabetic mellitus patients. METHODS: Neutrophils were isolated from SD rats and incubated in vitro. The cells were divided into four groups according to different concentrations of AGE in cell suspension: control group (C, with treatment of RPMI - 1640), A group (with treatment of 0.315 mg/mL AGE + RPMI - 1640), B group (with treatment of 0.625 mg/mL AGE + RPMI - 1640), D group (with treatment of 1.250 mg/mL AGE + RPMI - 1640). Activity of neutrophils were determined by MTT colorimetric assay. Selectin-L mRNA expressions were analyzed by reversible transcription polymerase chain reaction (RT -PCR) technique. The levels of reactive oxygen species (ROS) in neutrophils were measured with DCFH-DA method. The protein concentration of neutrophil elastase (NE) was assayed by ELISA. RESULTS: The activity of neutrophils were obviously increased in A, B, and D groups when compared with that in C group [(0.170 +/- 0.040) in C group, (0.320 +/- 0.030) in A group, (0.380 +/- 0.020) in B group, (0.290 +/- 0.010) in D group, P <0. 05]. The expression of Selectin-L mRNA in A, B, D groups were significantly higher than that in C group (0.95 +/- 0.08, 1.36 +/- 0.27, 0.50 +/- 0.26.vs.0.36 +/- 0.26, P < 0.05. respectively). The ROS levels in A, B, D groups was markedly higher than that in C group (1.64 +/- 0.20, 2.16 +/- 0.26, 3.26 +/- 0.75. vs. 0.72 +/- 0.15, P <0.05, respectively). The levels of NE in A, B, D groups were significantly increased when compared with that in C group(1.98 +/- 0.43, 2.50 +/- 0.43, 2.01 +/- 0.18 vs 0.91 +/- 0. 21, P <0.05, respectively). CONCLUSION: AGE can enhance the activity of neutrophil, with change in cellular biological behaviors, which may be one of main reasons for abnormal inflammation in wounds of diabetes mellitus patients. PMID- 18512552 TI - [Therapeutic effect of vacuum-assisted closure technology on infected explosion wound of pig]. AB - OBJECTIVE: To investigate the effect of vacuum-assisted closure (V. A. C.) technology in the treatment of infected wound of skin and soft tissue as a result of explosion injury in pig. METHODS: Sixteen explosion wounds were established by electric detonators on the shoulders and hips on both sides of 4 small white domestic pigs ,and they were divided into A group [(without treatment and infection occurred on 1-2 post burn day (PBD), then treated with vaseline gauze on 3 (PBD)], and B group (with the same treatment as in A group, except for treatment of vacuum assisted closure (V. A. C) with pressure of - 15 kPa after 3 PBD). The data of wound depth, wound area, wound healing time were collected and analyzed at 3 PAD and 1, 3, 6, 9, 14, 19, 24 days after treatment. Specimens from wounds were collected for histopathology observation, including also cell proliferation index, the number of vascular endothelial cells, the activity of myeloperoxidase (MPO) and the number of bacteria. RESULTS: Compared with those in A group on land 3 days after treatment, wound area, wound depth were not enlarged or deepened in B group, while the number of inflammatory cells, vascular endothelial cells, proliferative cells were increased, the activity of MPO was enhanced and the number of bacteria was decreased. There were obvious differences between two groups in following indices: wound area ,wound depth, the number of vascular endothelial cells and bacteria during 1 to 19 days after treatment (P < 0.01)), the number of cell proliferation from 1 - 9 days after treatment (P < 0.01)), and the activity of MPO on 3, 6 days after treatment (P < 0. 01). The wound healing time was (32.8 +/- 1.6) d in A group, which was longer than that in B group (25.8 +/- 1.0 d, P < 0.01). CONCLUSION: Compared with conventional dressing change, V. A. C can decrease bacteria load, lessen secondary necrosis, prompt the inflammatory response, accelerate the formation of granulation tissue, shorten wound healing time in infectious wound of porcine skin and soft tissue resulted from explosion injury. PMID- 18512553 TI - [Study of neovascularization disturbance in deep partial-thickness scald in rats with diabetes mellitus]. AB - OBJECTIVE: To study the relationship between the degree of neovascularization and non-healing wounds in scalded rats with diabetic mellitus. METHODS: Sixty Sprague Dawley rats were randomly divided into control group (C, n = 30, with treatment of isotonic saline) and streptozocin (STZ)-induced diabetic group (D, n = 30, with treatment of STZ), and then they were inflicted with 20% TBSA deep partial thickness scald. Wound specimens were harvested immediately after scald and on 1, 3, 7, 10, 14, 21 post scald days (PSD) to observe histological changes, and wound healing rates were calculated. Degree of neovascularization in wound (labeled with blue microsphere) and the quantity of vascular endothelial cells (labeled with red CD31) were also measured by double-labeling immunofluorescence. RESULTS: Compared with those in C group, Wound healing rate and histological value scores were lowered, and the degree of neovascularization was abated markedly at each time point. The degree of neovascularization in D group (12.00 +/- 1.40) was obviously lower than that in C group on 7 PSD (60.00 +/- 3.00, P <0.01). There was no obvious difference in the number of vascular endothelial cells in both groups, however, the majority of endothelial cells had not formed functional capillaries in D group. CONCLUSION: Vascular endothelial cell can proliferate actively with poor blood supply in diabetic nonhealing with deep partial thickness scald wounds, but it is still poor in blood supply due to lack of functional capillaries. PMID- 18512554 TI - [Effect of advanced glycosylation end products on cell cycle of epidermal keratinocyte and the role of signal pathway]. AB - OBJECTIVE: To investigate the effect of advanced glycosylation end products (AGE) on cell cycle of epidermal keratinocyte and its possible signal pathway. METHODS: 150 mg/L AGE-human serum albumin (AGE-HSA) was prepared in vitro. Primary cultured keratinocytes in logarithmic growth phase were harvested and divided randomly into: A group [with treatment of defined keratinocyte-SFM (DK-SFM) serum free medium], B group (with treatment of DK-SFM medium including 150 mg/L AGE HSA), C group (with DK-SFM medium after treatment of U0126) and group D (with D K SFM medium including 150 mg/L AGE-HSA after treatment of U0126). Cell cycle distributions were analyzed by flow cytometer. The protein levels of cyclin D1, cyclin B1, CDK4 and p44/42 MAPK were measured by Western blot. RESULTS: Compared with those of A group, the percentage of S-phase and G2/M-phase keratinocytes were decreased obviously in B group, the percentages of G2/M -phase keratinocytes showed the same tendency in C and D groups [(9.7 +/- 1.1)% , (9.8 +/- 0.7)%, respectively, P <0.05]. Compared with that of A group, the expression of cyclin D1 were decreased significantly in other groups, among which a weak expression was showed in D group. There was no obvious difference between A and B groups in CDK4, or cyclin B1 and p44/42 MAPK protein levels ,which were significantly higher than those in C and D groups. CONCLUSION: AGEs inhibit the progress of cell cycle of keratinocytes by downregulation of cyclin D1 expression. PMID- 18512555 TI - [Clinical application and long-term follow-up study of acellular dermal matrix combined with auto-skin grafting]. AB - OBJECTIVE: To investigate the clinical effect of acellular dermal matrix (ADM) combined with auto-skin grafting on deep burn wound ,and the result of long-term follow-up and histological examination. METHODS: One hundred and fifty-two patients with deep burn hospitalized from February 2000 to July 2003 were repaired with porcine ADM and auto split-thickness graft. Wound healing rate was assessed 1 week after operation. Degree of cicatricial hyperplasia was examined 1, 3, 6, 12 months after operation. Wound samples from 5 patients were harvested for histological examination 72 months after operation, for which transmission electron microscopy were employed in 2 cases. RESULTS: Grafts completely survived was seen in 116 patients (accounting for 76.3% of cases), survival rate over 95% were observed in 23.7% of cases. One hundred and twenty-seven patients were followed up 1 month after operation, in whom mild local contraction, cord like scar was seen along its junction with skin, its texture was soft ,and there was no pruritus or blister formation. One hundred and one patients were followed up 3 months after operation, and the graft showed mild contraction less marked when compared with that of the site where auto split-thickness skin grafting was used. Articular function was good. Eighty-two patients were followed up 6 months after operation,color and texture of grafts were similar to normal skin with no obvious cicatricial hyperplasia. Fifty-eight patients were followed up 12 months after operation, the texture of grafts was similar to normal skin without obvious reject reaction. Sixteen patients were followed up over 72 months after operation, the grafts appeared dry compared with normal skin. Histological examination showed: tissue structure of grafts was similar to normal skin, intact small sweat gland and sweat gland cells were not found in dermal layer. CONCLUSION: Heterologous ADM combined with auto split-thickness graft can survive in human body without obvious immune rejection reaction for a long time. No intact small sweat gland or sweat gland cells in dermis is a problem worth of study in regeneration of skin function. PMID- 18512556 TI - [Repair of high-voltage electrical burn in the neck]. AB - OBJECTIVE: To explore methods of repair of high-voltage electrical burn in the neck. METHODS: Thirty-seven patients with high-voltage electrical burn in neck hospitalized since 1985 were enrolled in this study. After debridement, the wounds were repaired with latissimus dorsi myocutaneous flap, trapezius myocutaneous flap, platysma myocutaneous flaps, pectoralis major myocutaneous flap, or latissimus dorsi myocutaneous flap combined with pectoralis major myocutaneous flap. RESULTS: Necrosis occurred at edge of flap (about 1 - 2 cm in breadth) in 3 patients, and the other flaps survived well with perfect appearance and local function. CONCLUSION: To repair with pedicled myocutaneous flaps and combined flaps after early debridement can be safe, effective and reliable in the management of patients with high-voltage electrical burn in the neck. PMID- 18512557 TI - [Pharmacokinetics changes of amikacin in severe burn patients at early stage]. AB - OBJECTIVE: To investigate the concentration and pharmacokinetics changes of amikacin in the serum and blister fluid in severe burn patients at early stage. METHODS: Twenty severe burn patients during early postburn stage were divided into four groups with five patients in each group. Each patient was given a single dose of 400 mg amikacin in 30 minutes during 3-4 postburn hour (PBH) in A group, at 10 PBH in B group, at 20 PBH in C group, and at 30 PBH in D group. The concentration of amikacin in blister fluid was examined at 0.25, 0.5 min and 1, 2, 3, 4, 5, 6, 7 h after treatment by fluorescence polarization immunoassay, meanwhile, the venous blood of 9 patients among them was also collected to determine the concentration of amikacin at the same time points. Pharmacokinetics parameters of model were produced by program 3P97. RESULTS: Among all groups, the concentration of amikacin in blister fluid in A group increased quickest and maintained longest, that of B group ranked second. The amikacin concentration of blister fluid in A, B groups were obviously higher than those in C, D groups at each time point (P <0.05 orP < 0.01), especially at 1PBH (12.53 +/- 1.76, 9.52 +/ 1.51 microg/mL vs 4.65 +/- 0.77, 3.10 +/- 0.41 microg/ml, P < 0.01). The serum concentration of amikacin in 9 patients were decreasing along with elapse of time. The amikacin concentration-time curves in blister fluid and serum were best fit in two compartment models. Compared with that in normal value, t1/2beta of amikacin from burn patient was shortened in serum and prolonged in blister fluid. CONCLUSION: Early administration of amikacin in burn patients (within 10 PBH) may form an effective and continuous antibiotics barrier around the wound to prevent bacterial infection. PMID- 18512558 TI - [Rescue and treatment for the mass burn casualties of yellow phosphorus explosion]. AB - OBJECTIVE: To summarize the characteristics and treatment of burn casualties of yellow phosphorus explosion, so as to share the experiences in emergency treatment. METHODS: By analyzing the data related to this accident, the characteristics of the injury and experiences of treatment for mass burn casualties from yellow phosphorous explosion were summarized. RESULTS: Eighty-one patients, 72 males and 9 females, were injured in a yellow phosphorus explosion. The mean age of the patients was 24 +/- 13 years old (5-42 y). The mean total burn surface area was (9 +/- 11)% [(0.4% - 70.0%))] TBSA, and the mean burn surface area of III degrees/IV degrees was (7 +/- 10)% [(0.4% - 60.0%)] TBSA. Most of the patients showed the symptoms and signs of phosphorus poisoning. Among all the patients, 27 cases (33.3%) showed hepatic dysfunction, 15 cases (18.5%) had renal dysfunction, 42cases (51.9%) showed electrolytes disorders. Among the 8 patients with burn surface area over 10% TBSA and less than 20% TBSA, high levels of cardiac enzymes were found in 6 cases, anaemia in 7 cases (3 with progressive anaemia), asphyxia occurred in 1 case 48 hours after burn, and in 1 case complicated with stress ulcer. Escharectomy and skin grafting were performed within four days after burn in 72 patients. All the patients survived, some of them showed impaired hand function and hypertrophic scar, and partial finger amputation was done in 3 patients. CONCLUSION: Yellow phosphorus explosion produces deep burn injuries in surrounding people especially in exposed parts such as head, hand and so on. Adequate organization of medical resources for emergency treatment, early debridement, and accelerating excretion of phosphorus are the key points for the successful rescue of mass casualties. PMID- 18512559 TI - [Preliminary investigation on the dynamic change in epidermal stem cells under mechanical stress in vivo]. AB - OBJECTIVE: To observe the distribution of epidermal stem cell (ESC) after soft tissue expansion, and to explore dynamic change in ESC under mechanical stress and kinetic mechanism of skin expansion. METHODS: Skin samples were collected from patients after expansion of the scalp. They were divided into three groups: A group (scalp harvested 3 cm away from the center of dilator), B group (scalp tissues at the edge of dilator), and control group (scalp without dilatation). The tissue structures were observed with optical microscope with HE staining. The distribution and differentiation characteristics of cell keratin 19 (CK19) positive cells were observed with inverted phase contrast microscope after immunohistochemistry staining. RESULTS: HE staining showed that the epidermis was thickened and distributed densely with uneven, rugged and increased layers in A, B groups. With immunohistochemistry staining, CK19 positive cells appeared in multilayers in basal membrane, a few of them were in cluster or dispersed , with" hollowing" structure formation. These phenomena were not seen in control group. CONCLUSION: ESC can proliferate with abnormal distribution and "hollowing" structure formation after mechanical dilatation, which may be related to dynamic changes in basal layer cells. PMID- 18512560 TI - [Experimental study on gelatin/polycaprolactam composite nanofiber scaffold in wound healing]. AB - OBJECTIVE: To explore the effects of the gelatin (Gt)/(polycaprolactam) PCL composite nanofiber scaffold on wound healing of full-thickness defect in rabbits. METHODS: Sixteen rabbits were inflicted with full-thickness skin open wounds on the back, and they were divided into A group (with treatment of Gt/PCL nanofiber membrane, 8 wounds), B group (with treatment of PCL fiber membrane, 8 wounds), and C group (with treatment of vaseline gauze, 8 wounds), the wounds in A , B groups served as self controls. Wound healing time was recorded. Wound healing rates were calculated on 3, 7, 10 post operation day (POD). Wound specimens and their adjacent skin were examined histologically on 3, 7, and 10 POD. RESULTS: There was obvious difference in wound healing time among A, B, C groups, and they were (18.2 +/- 1.3) d, (20.3 +/- 1.1) d, (22.0 +/- 0.6) d, respectively. Wound healing rate in A group was obviously higher than that in B, C group on 3, 7, 10 POD (P < 0.05). Compared with those in other groups, the proliferation of granulation tissue in dermis in A group was less, the epidermal cells were proliferated quickly, and collagen was arranged regularly. CONCLUSION: Gt/PCL composite nanofiber membrane can promote wound healing of full-thickness skin defect in rabbits, which is an ideal material for tissue engineering. PMID- 18512561 TI - [The expression of lefty protein in adult normal skin, human embryonic skin and hyperplastic scar]. AB - OBJECTIVE: To observe the expression of lefty in adult normal skin (ANS), human embryonic skin (HES) and hyperplastic scar (HS), and to explore the effect of lefty on HS and the relationship between lefty and scarless wound healing in embryo. METHODS: Samples of ANS, HES and HS were collected for frozen section for immunofluorescence staining. The morphology of fibroblast and the expression of the lefty were observed by laser confocal microscopy, and the positive cell rates were calculated. RESULTS: Fibroblasts in ANS and HS were long and fusiform with regularity, their nuclei were fusiform or stellate and irregular. Fibroblasts in HES were fusiform, while nuclei were elliptic or fusiform and regular. Positive cell rates of lefty protein in HS (15.38%) were lower than that in NS (67.92%) and FS (81.67%, P < 0.01), and it was lower in ANS compared with HES (P <0.05). CONCLUSION: Lefty protein may inhibit the formation of scar, its high expression may be related to the embryo scarless wound healing. PMID- 18512562 TI - [Influence of macrophages on some biological features of endothelial cells]. AB - OBJECTIVE: To establish the co-culture model of human macrophage cell line (U937) with human vein umbilical cell line (ECV304), and to explore the feasibility of using concanavalin A (ConA) as U937 cell stimulator in regulating angiogenesis. METHODS: ECV304 cells were cultured in vitro, and to which were respectively added U937 cells (1 x 10(5)), 25 microg/mL ConA, and U937 cell (1 x 10(5)) + ConA (25 microg/mL) after cell fusion rate reaching 60%, and then co-cultured for 48 hours. ECV 304 cells in conventional culture were used as controls. 3H-TdR incorporation test was employed to determine the DNA synthesis of vascular endothelial cells. Flow cytometry was used to determine the changes in the cell cycle, and RT-PCR was adopted to determine the expression of homeobox (HOXB2) mRNA. RESULTS: After conA stimulation to ECV 304 co-cultured with U937 cells, the percentage of cells in S phase (48.860 +/- 2.290), the DNA synthesis [(5694 +/- 917) min(-1)], and the expression of HOXB2 mRNA (0.947 +/- 0.003) were obviously higher than those in control group [41.590 +/- 2.590 vs (2498 +/- 1109) min(-1) vs 0.646 +/- 0.004, P > 0.01]. There was no obvious difference in apoptosis among above stimulation methods (P >0.05). CONCLUSION: U937 cells activated by ConA can promote the proliferation of ECV304 cells and further regulate angiogenesis. HOXB2 gene is closely related to the endothelial proliferation. PMID- 18512563 TI - [Quantification of type I and III collagen content in normal human skin in different age groups]. AB - OBJECTIVE: To quantify the content of type I, III collagen and their ratio in normal human skin of different age, and to explore the regulation of changes. METHODS: The normal human skin specimens were obtained from 6 spontaneously aborted fetus and 56 burn patients of different ages, including infants (newborn 3 years), pre-school group ( > 3, < or =7 years), adolescent group ( >7, < or = 18 years), youth and middle age group ( > 18, < or = 50 years), and elderly group ( > 50 years), were studied. The total collagen content were determined by hydroxyproline method. The contents of type I, Ill collagen and their ratio were examined by immunohistochemistry. RESULTS: The total collagen content decreased along with increase in age, and it was highest in fetus [(543 +/- 13) microg/g]. The ratio between type I and Ill collagen increased along with increase in age. The content of type III collagen was highest in fetus [(278 +/- 7) microg/g], and it decreased along with increase in age. The content of type I collagen content was [(265 +/- 7) microg/g] in fetus, and it was increased slightly in infant and pre-school groups, then decreased along with advance in age. CONCLUSION: Decomposition of type III collagen in normal human skin may exceed its synthesis after birth immediately, leading to its reduction. Synthesis of type I collagen in normal human skin is dominant before 8 years old, and it shows an opposite tendency afterwards. PMID- 18512564 TI - Narrative research--learning from stories. PMID- 18512565 TI - Supportive care framework. AB - The Supportive Care Framework for Cancer Care was originally formulated in 1994 (Fitch, 1994). The framework was designed as a tool for cancer care professionals and program managers to conceptualize what type of help cancer patients might require and how planning for service delivery might be approached. The framework has been presented in various arenas and the number of requests has been growing for a wider distribution of a full description of the framework. The purpose of this article is to share the Supportive Care Framework for Cancer Care with the cancer nursing community. As a conceptual framework, it may be a useful tool for service or program planning, a basis to organize educational approaches in cancer care, or as a model underpinning research projects. PMID- 18512566 TI - Introducing a patient-focused care map in colorectal surgery: a pilot qualitative study of patients' and surgical oncology nurses' experiences. AB - Care maps for patient care have been around for many years. Key stakeholders at our institution developed and implemented a care map for patients undergoing surgery for colorectal cancer. The purpose of this descriptive, qualitative pilot study was twofold. First was to understand the lived experience of patients being cared for under a newly-implemented care map utilizing patient diaries and interviews. The second goal was to describe the experiences of surgical oncology nurses caring for these patients using a focus group technique. The results of our small study indicated that patients appreciated having a document that outlines daily activities and goals, and were anxious to get home, but were disappointed in the discharge planning process. Nurses were positive about the care map overall, but felt they could have contributed more in the development and planning stages of the care map. Overall, the implementation of our patient centred care map was a success. PMID- 18512567 TI - Reflecting on spirituality in the context of breast cancer diagnosis and treatment. AB - In this first part of a longitudinal study, women were asked to reflect on the meaning of spirituality in the first year following diagnosis of breast cancer. Twenty-two women were interviewed at approximately one year post-diagnosis. This paper reports on a thematic analysis of these interviews. Participants' responses reflected three higher-order themes: relationship with a higher power, a deepening sense of self, and spiritual connection with others. The findings provide an enhanced understanding of how spirituality frames and impacts (both positively and negatively) the experience of breast cancer immediately following diagnosis and treatment. Most participants in this study found strength and support in their experiences of spirituality. They also spoke at times of feeling disconnected from or abandoned by God. The paper concludes with a discussion of how cancer health professionals might respond to the spiritual needs expressed by women living with cancer. PMID- 18512568 TI - Living after cancer: challenges in being a survivor. AB - Concerns about survivorship and the needs of cancer survivors are occurring with increasing frequency (Ferral, Virani, Smith & Juarez, 2003; Curtiss & Haylock, 2006). Advances in the diagnosis and treatment of cancer have resulted in an ever growing cadre of individuals who are survivors of the disease. In United States alone, there are more than 10 million cancer survivors (ACS, 2005). In Canada, there are almost 800,000-a comparable number given the country's population (NCIC, 2006). Approximately 60% of adults who are diagnosed with the disease and 78% of the children are alive at five years (ACS, 2005). Given the expectation that the number of people diagnosed with cancer will double in the next 40 years, we can expect the number of survivors will also continue to increase. Living after a diagnosis of cancer and its subsequent treatment is not without its challenges. We are only now beginning to recognize some of the concerns and issues survivors face and what a vulnerable population these individuals constitute. The growing number of individuals in our midst has allowed us to start learning about the challenges survivors can face on a daily basis. Their voices are being heard as advocacy group representatives speak out about their needs and the gaps in cancer service delivery. We are beginning to identify the spectrum of late complications survivors may experience with the potential to compromise quality of life. We are also beginning to recognize that the late and long-term effects are more prevalent, serious, and persistent than was originally expected. PMID- 18512569 TI - Mentorship: off and running to improve dyspnea in lung cancer patients. PMID- 18512570 TI - Where would we be without mentors? PMID- 18512571 TI - Remembering Mrs. Chase. PMID- 18512572 TI - Going for the gold: credit bearing independent studies hold academic treasure. PMID- 18512573 TI - Incivility in nursing education. PMID- 18512574 TI - Learning to ask for help. PMID- 18512575 TI - Embracing mentoring in faculty relationships. PMID- 18512576 TI - Experiencing an externship. PMID- 18512577 TI - More on peer review. PMID- 18512578 TI - SWOT is useful in your tool kit. AB - The components of SWOT are addressed in this column as a methodology to identify a department's strengths, weaknesses, opportunities, and threats. PMID- 18512579 TI - Transition from hospital to home care: what gets lost between the discharge plan and the real world? AB - Hospital-based nurses who have not practiced in home health care may find it difficult to anticipate patients' needs during the transition from hospital to home. This column focuses on increasing competency in preparing patients for home health services. PMID- 18512580 TI - Continuing professional competence: peer feedback success from determination of nurse locus of control. AB - Continuing competence programs (CCPs) are becoming required in many nursing jurisdictions across Canada. CCPs have the common goal of ensuring nurses remain up-to-date and competent in their professional practice. They often incorporate reflective practice as a tool for nurses to critically explore their practice and professionalism on an ongoing basis. Feedback is one of the strategies within reflective practice which enables nurses to increase reflective capabilities and enhance professional performance through collegial exchange. Locus of control, a personality variable, is considered as a potential indicator of how receptive nurses may be toward CCPs. For CCPs to be optimally embraced by nurses, it is imperative that professional nursing bodies address nurses' loci of control in developing, supporting, and strategizing for reflective practice, generally, and feedback, specifically. PMID- 18512581 TI - Precepting in the fast lane: improving critical thinking in new graduate nurses. AB - BACKGROUND: Research regarding the relationship between preceptorship and development of critical thinking in new graduate nurses is sparse. No studies could be found that examined the relationship of preceptor education to critical thinking scores of new graduate nurses. METHODS: A quasi-experimental, mixed methods design measured critical thinking ability of new graduate nurses. Focus group interviews were conducted with preceptors who attended an author-developed educational program. RESULTS: Preceptors' participation in the educational session contributed to the evaluation subscale of critical thinking skills of the experimental group on the California Critical Thinking Skills Test (F = 4.709, p = .039). Preceptors had positive qualitative responses. CONCLUSIONS: Critical thinking skills of new graduate nurses can be improved and learning relationships developed through preceptor education. Further study is suggested. PMID- 18512582 TI - An academic service partnership to expand capacity: what did we learn? AB - This article highlights the educational needs that emerged when an academic institution and a hospital partnered to produce more nurses in response to the registered nurse shortage. The partnership proposed to offer quality clinical experiences for students by having the hospital-paid nurse clinicians serve as faculty. This would enable the academic partner to increase enrollment amid limited faculty resources. The qualitative feedback received may prove useful in (1) guiding the professional development activities of future nurse clinicians who participate in education-service partnerships to expand educational capacity; (2) assisting academic nursing program administrators and faculty to expand capacity without compromising quality; and (3) assisting hospital nurse administrators to weigh risks and returns to pursue mutually beneficial outcomes. PMID- 18512583 TI - JCAHO preparation: an educational plan. AB - This article discusses Joint Commission on Accreditation of Healthcare Organizations (JCAHO) preparedness. A literature search reveals articles discussing varying tactics for addressing JCAHO preparedness (e.g., mock surveys, crossword puzzles, e-mail and paper updates, games, and pocket resource cards). However, no articles address the use of monthly pretests and posttests. This article focuses on the use of pretests and posttests as well as other interventions to prepare staff for the JCAHO tracer methodology. PMID- 18512584 TI - Consumer-directed teaching of health care professionals involved in the care of people with spinal cord injury: the Consumer-Professional Partnership Program. AB - The Consumer-Professional Partnership Program (CPPP) was developed to increase patient participation in the education of nurses and other health care professionals. Its goal is to inform nurses about disability and health issues from the perspective of people with spinal cord injury. This article discusses the development of this program and the first experiences with its implementation. Forty-three nursing professionals took part in CPPP training. Initial participant responses were positive, especially with regard to hearing from consumers about care preferences, barriers to care, and ways to improve patient-provider interaction. PMID- 18512586 TI - Ready for primetime? The desire for sophisticated cardiovascular information systems may exceed what's on the market today. PMID- 18512585 TI - Using technology to build community in professional associations. AB - Professional associations increasingly rely on technology to convene members in the conduct of their business. Principles of adult education can be employed to facilitate the transition to the use of innovative technology. Such innovation can facilitate an association's work in advancing policy and professional practice. Understanding generational differences can enhance the development of community and build social capital in associations. PMID- 18512587 TI - Turning crisis into opportunity. When Christ Hospital separated from its parent, the chance to build a first-class IT organization came into focus. PMID- 18512588 TI - The power of EMPI. Health systems are tapping the value of EMPIs to eliminate duplicate patient records. PMID- 18512589 TI - Trouble at the bedside. Nurses are fed up with wireless POC applications that just aren't reliable, and they're not going to take it anymore. PMID- 18512590 TI - Imaging invades surgery. As imaging technologies become more intertwined with the OR, CIOs must have a sound strategy for how the two will meet. PMID- 18512591 TI - A mandate in the making? As e-prescribing talk swirls around the halls of congress, the industry braces for a fiat. PMID- 18512592 TI - Future state. Newt Gingrich's vision of tomorrow's healthcare system involves copious amounts of IT. Interview by Anthony Guerra. PMID- 18512593 TI - The art of the project. Healthcare CIOs are under enormous pressure to manage a growing portfolio of ultra-complex projects. PMID- 18512594 TI - Getting smart. Looking to avoid allergic reactions and increase patient safety, UPMC creates some very smart rooms. PMID- 18512595 TI - Writing a new Chapter. A key joint venture between anesthesiologists and OR nurses is brought into focus with pre-op, inter-op and post-op care and documentation. PMID- 18512596 TI - [Practice during bombing. Interview by Charles Meunier]. PMID- 18512597 TI - [An ambassador for the profession. Interview by Marie Clark and Jean-Marie Papineau]. PMID- 18512598 TI - [Advocating for the elderly. Interview by Marie Clark]. PMID- 18512599 TI - [Special units. Interview by Charles Meunier]. PMID- 18512600 TI - [On the pathway. Interview by Charles Meunier]. PMID- 18512601 TI - [An added value. Interview by Charles Meunier]. PMID- 18512602 TI - [A wheel that turns. Interview by Veronique Robert]. PMID- 18512603 TI - [Poor quality of life. Interview by Jeanne Morazain]. PMID- 18512604 TI - [Long live rooming-in]. PMID- 18512605 TI - [Diversity in all directions. Interview by Jean-Marc Papineau]. PMID- 18512606 TI - [Journey of the combatants. Interview by Veronique Robert]. PMID- 18512607 TI - [Measuring the indices]. PMID- 18512608 TI - [To your keyboards!]. PMID- 18512610 TI - Neonatal options. PMID- 18512612 TI - When the State Board calls: part I guidance from nurse attorney, Latonia Denise Wright. PMID- 18512614 TI - Moral distress in healthcare practice: the situation of nurses. PMID- 18512615 TI - Committe identifies significant trends in review of conduct decisions. PMID- 18512616 TI - Lack of standard dosing methods contributes to i.v. infusion errors. PMID- 18512617 TI - An RN shares her perspective on forensic psychiatric nursing. PMID- 18512618 TI - Composite restoration of enamel defects. PMID- 18512619 TI - Editorship: something like "pulling teeth"! PMID- 18512620 TI - Comparison of cold GP/sealer and resin bonded obturation techniques in canine teeth in dogs. AB - An in vitro study compared two obturation materials in the canine teeth in dogs. The teeth were instrumented with rotary instruments and obturated with either gutta percha and a sealer or resin-based materials, utilizing the gutta percha apical plug/master cone technique. Radiographs were used for evaluation of the overall appearance of the finalfill. A modified apical dye leakage method was used to evaluate the ability of each material to provide an adequate barrier to apical leakage. When comparing the two obturation materials, the differences in the radiographic appearance scores and the apical dye leakage test results were not statistically significant. The rate of apical leakage is comparable to other obturation methods that have been reported. The use of a rotary system in conjunction with the gutta percha apical plug/master cone technique is valid for the endodontic treatment of indicated teeth. PMID- 18512621 TI - Locked jaw syndrome in dogs and cats: 37 cases (1998-2005). AB - A consecutive series of cases of dogs and cats with locked jaw syndrome (inability to open or close the mouth) are reported in this study. Dogs were significantly overrepresented (84.0%) and adult dogs were more frequently affected (81.0%). Temporomandibular joint ankylosis due to fracture was the most common cause (54.0%) of locked jaw syndrome. Additional potential causes of locked jaw syndrome are masticatory muscle myositis, neoplasia, trigeminal nerve paralysis and central neurological lesions, temporomandibular joint luxation and dysplasia, osteoarthritis, retrobulbar abscess, tetanus, and severe ear disease. Treatment of locked jaw is directed towards the primary cause. It is important to treat the tonic spasm in order to minimize periarticular fibrosis. Surgical intervention is recommended for temporomandibular joint ankylosis. Masticatory muscle myositis treatment is initiated by gradually opening the mouth, with medical treatment based on immunosuppressive therapy. Fracture and masticatory muscle myositis are associated with a relatively good prognosis in regard to short-term outcome as compared to animals with central neurologic lesions or osteosarcoma which have a poor prognosis. PMID- 18512622 TI - Evaluation of cross-protection by immunization with an experimental trivalent companion animal periodontitis vaccine in the mouse periodontitis model. AB - Companion animal periodontal disease is one of the most prevalent diseases seen by veterinarians. The goal of this study was to evaluate the vaccine performance of a trivalent canine periodontitis vaccine in the mouse oral challenge model of periodontitis. Mice vaccinated subcutaneously with an inactivated, whole-cell vaccine preparation of Porphyromonas denticanis, Porphyromonas gulae, and Porphyromonas salivosa displayed significantly reduced alveolar bone loss in response to heterologous and cross-species challenges as compared to sham vaccinated animals. Based on the results of these studies, a periodontitis vaccine may be a useful tool in preventing the initiation and progression of periodontitis caused by the most commonly isolated pigmenting anaerobic bacteria in animals. PMID- 18512623 TI - Effect of a mucoadhesive gel and dental scaling on gingivitis in dogs. AB - Twenty client-owned dogs diagnosed with gingivitis were studied over a 45-day period in order to investigate the effect of professional dental prophylaxis combined with the use of a topical mucoadhesive gel containing adelmidrol, an aliamide. A non-intrusive papillary-marginal-gingival index (PMGI) was measured at each assessment, while the gingivitis index (GI) was measured only at the beginning and end of the study. Compared to the control group, the treated dogs had a significant decrease (P < 0.005) in the average GI index during the course of the study. A significant reduction (P < 0.002) in the average PMGI index was observed in both groups 15-days following dental prophylaxis. However at 30 and 45-days following dental prophylaxis, the PMGI index values were significantly different (P < 0.005) from baseline only in treated dogs. These results suggest that the combined use of a mucoadhesive gel with dental scaling was able to improve the regression of gingival inflammation and lengthen the therapeutic benefits of dental scaling and polishing during a limited study period. PMID- 18512624 TI - Dental bulge restoration and gingival collar expansion after endodontic treatment of a complicated maxillary fourth premolar crown-root fracture in a dog. AB - This case report describes endodontic, restorative, and periodontal treatment of a complicated crown-root fracture of the right maxillary fourth premolar tooth in a dog. A buccal portion ('slab') had separated from the tooth, which extended subgingivally into root structure. Following completion of standard root canal therapy, a periodontal flap was elevated. Alveolectomy and alveoloplasty were performed, and inflamed soft tissues were debrided. The fracture site was prepared, restored, and shaped to receive a dental bulge contour. A gingival collar expansion technique was utilized to allow for apical positioning of gingiva at the distobuccal crown-root segment. Clinical and radiographic examination 15-months following treatment showed no evidence of endodontic failure. The restorations were intact, and periodontal probing depths were slightly increased probably due to gingival enlargement from concurrent cyclosporine therapy. The importance of biologic width maintenance in periodontal surgery and need for owner compliance with home oral hygiene are discussed. PMID- 18512625 TI - The periosteal releasing incision. PMID- 18512626 TI - Extraction of teeth in the mandibular quadrant of the cat. PMID- 18512627 TI - WANTED: clinical experts to teach the next generation of gerontological nursing professionals. Why it's important to be an "early adopter" of the Doctor of Nursing Practice. PMID- 18512628 TI - May Futrell, PhD, FAAN, FGSA. Gerontological nursing and gerontology leader and mentor. PMID- 18512629 TI - Validating MDS data about risk factors for perineal dermatitis by comparing with nursing home records. AB - Perineal dermatitis is one of the main complications of incontinence and increases the cost of health care. The Minimum Data Set (MDS) contains data about factors associated with perineal dermatitis identified in a published conceptual model of perineal dermatitis. The purpose of this study was to determine the validity of MDS data related to perineal dermatitis risk factors by comparing them with data in nursing home chart records. Findings indicate that MDS items defining factors associated with perineal dermatitis were valid and supported use of the MDS in further investigation of a significant, costly, and understudied health problem of nursing home residents. PMID- 18512630 TI - Social support as a moderator in a fall prevention program for older adults. AB - This study examined the moderating effects of social support in a fall prevention program for community-dwelling older adults. Twenty-six Japanese older adults age 65 and older participated in a 2-month exercise program and were measured in anthropometrical, physical, and psychological functioning at baseline, 2 months (intervention termination), and 5 months (3 months after intervention termination). Analyses indicated that the program did not improve participants' body mass index, balance, or walking speed. However, participants'falls self efficacy significantly increased from baseline to intervention termination and was maintained at a higher level at the 5-month postintervention follow up. This efficacy-improving effect was prominent in the participants who had received less social support at baseline. The findings emphasized the importance of considering participants' social resources and targeted intervention outcomes when evaluating the effect of exercise. PMID- 18512631 TI - End-of-Life Nursing Education Consortium. Geriatric Training Program: improving palliative care in community geriatric care settings. AB - Recent studies of end-of-life care in nursing homes and other long-term care settings point to a significant need to improve care. The End-of-Life Nursing Education Consortium (ELNEC)-Geriatric Training Program is an important educational initiative to advance palliative care and end-of-life education for licensed nurses and nursing assistants. The ELNEC-Geriatric Training Program prepares nurses as educators and leaders to improve the quality of end-of-life care in geriatric care facilities. This article presents evaluation data from the 2007 pilot ELNEC-Geriatric Training Program and follow-up evaluation of the "train-the-trainer" model to disseminate comprehensive palliative care education in geriatric settings. PMID- 18512632 TI - Korean older adults' perceptions of the aging process. AB - This qualitative study was conducted to describe Korean older adults' perceptions of the aging process. A total of 18 Korean older adults were interviewed, and a grounded theory approach was used to analyze the interview data. The participants were found to perceive aging as a process of Generating, Expressing, and Transforming of Growing Futility. The degree to which they perceived their Growing Futility depended on the actions and interactions of a set of conditional structures. This study revealed five patterns of Korean older adults' perception of the aging process. These findings allow for the possibility of a more refined theoretical development for the aging process, especially when a comparative study becomes available through cross-cultural qualitative research. PMID- 18512633 TI - Implementation and effects of a medication pass nutritional supplement program in a long-term care facility: a pilot study. AB - The purpose of this 6-month study was to determine the most effective method for implementing an interdisciplinary medication pass nutritional supplement program in Extendicare's long-term care facilities in western Canada by piloting the program in one facility. The program's effects on residents' body weight, skin integrity, and laboratory values related to nutritional status were evaluated. Resident adherence and staff perception were also assessed. Findings indicated that most residents experienced weight gain and improved visceral protein status. Resident adherence to the program was 96%, and staff feedback was positive. The promising results of this pilot study suggest that a medication pass nutritional supplement program could help improve the nutritional status of long-term care residents, thereby resulting in improved quality of life. PMID- 18512634 TI - Re-education about erectile dysfunction. PMID- 18512635 TI - Managing tinnitus. AB - Tinnitus affects at least one in 10 people at some time in their lives. They should consult their general practitioner so that any underlying causes can be assessed, including hearing loss. Patients who find tinnitus affects daily life and work or personal relationships can be helped by strategies such as sound therapy devices, relaxation exercises or joining a self-help group. There is a need to raise awareness of tinnitus among health professionals, the public and patients and their families. Surveys of tinnitus patients suggest that they fee isolated and that their families and employers have little understanding of the condition. Voluntary organisations such as the Royal National Institute for Deaf People (RNID) and the British Tinnitus Association provide useful information and resources for lay people and health professionals. PMID- 18512636 TI - The nursery nurse: time to change the title? AB - Nursery nurses have an important role to play in community child health services by promoting and maintaining family health. The title of "nursery nurse", however, does not reflect their knowledge or their skills. PMID- 18512637 TI - Low vitamin D status: on the increase? AB - Maximising bone strength in childhood and adolescence is critical for future bone health and reduction in osteoporosis. Vitamin D plays a key role in bone development, bone mineral density and is essential to maximise the absorption of calcium from the gut. In the UK it has been estimated that exposure to sunlight is sufficient to maintain adequate vitamin D status, hence no dietary reference value is set for the majority of the population. However, the prevalence of vitamin D deficiency and rickets is increasing and moderate-to-mild vitamin D deficiency has been demonstrated to produce ower bone mass in children and adolescents, placing them at risk of osteoporosis in later life. This paper argues that the UK should set dietary reference values for vitamin D for the whole of the population, in line with other Western countries, and give greater emphasis to dietary sources of vitamin D, including through fortification of foods. PMID- 18512638 TI - Fabrics for atopic dermatitis. AB - The type of fabric worn by sufferers from atopic dermatitis should not exacerbate the condition but, if possible, help to control it. Synthetic fabrics and wool tend to produce itching and irritate the skin. Cotton is traditionally recommended but its structure contains short fibres which expand and contract, causing a rubbing movement that can irritate delicate skin. Dyes used in cotton garments can increase the potential of a sensitivity reaction. Cotton is also prone to bacterial and fungal attack. Silk garments are often closely woven which impedes the flow of air, and some people are allergic to the sericin protein in silk. Published studies suggest that a specially treated silk material (DermaSilk), which is loosely knitted, has had the sericin removed and has a microbial agent (AEM 5772/5) permanently bonded to it, is well tolerated and has beneficial effects on the skin of children and adults with atopic dermatitis. Atopic dermatitis often becomes infected, commonly with Staphylococcus aureus. Some studies have investigated the use of clothing materials impregnated with substances such as silver, which has antimicrobial properties. However, these are still unproven and there are concerns about bacterial resistance and the local and environmental effects of silver. The use of the antimicrobial AEM 5772/5, which does not transfer to the skin of the patient, is a new development in the control of atopic dermatitis. Further studies are needed to determine whether an antimicrobial shield bonded to clothing material will reduce the colonisation of atopic skin by S. aureus. PMID- 18512639 TI - The clinical use of probiotics for young children. AB - Probiotics are live microorganisms that occur naturally in the human digestive system. They chiefly colonise the colon and benefit the health of the individual by acting as a digestive aid and boosting the immune response. Low levels of beneficial bacteria and microorganisms in the colon provide a greater opportunity for pathogenic bacteria to become established. Probiotics may be beneficial in reducing the severity and incidence of gastroenteritis, diarrhoea, and colic in babies. They can also benefit children's health by supporting the developing immune system but should not be seen as a panacea or as an alternative to a healthy lifestyle or diet. The level of probiotics in the colon can be boosted by a healthy diet containing plenty of grains, fruit and vegetables. It can also be boosted by probiotic supplementation. Supplementation by the probiotic L. reuteri has been studied in major scientific trials and proven to be safe and effective with no reported side effects. PMID- 18512640 TI - Parent-infant relationships and the role of the health visitor. AB - This article looks at research on emotional development in young children and how this is influenced by the attachments they make with their primary carers. The paper also considers the pivotal role health visitors might have in identifying and supporting families where there are early parent-infant relationship problems. PMID- 18512641 TI - Conversion disorder mimicking serotonin syndrome in an adolescent taking sertraline. AB - We present the case of a 15-year-old-boy on sertraline (Zoloft) who presented to the Emergency Department with palpitations, tremor, and weakness after starting erythromycin. Upon admission to the hospital, he developed syncopal-like episodes of unresponsiveness associated with body-wide jerking and stiffening as well as tachycardia and hypertension. He was initially believed to have serotonin syndrome and was transferred to a pediatric Intensive Care Unit where the diagnosis of conversion disorder was made. We discuss the salient features of serotonin syndrome and conversion disorder and emphasize the importance of making either diagnosis. PMID- 18512642 TI - The founders of the Medical Society of Delaware: Doctor Joseph Philippe Eugene Capelle. PMID- 18512643 TI - Maverick doc. PMID- 18512644 TI - Four stitches in time. PMID- 18512645 TI - Doc, I'm in the donut hole. PMID- 18512646 TI - IBM investigations--when has a doctor been "investigated?". PMID- 18512647 TI - Conflict of interest policies under review at the UI. PMID- 18512648 TI - Medical staff bylaws in Iowa--a contract? PMID- 18512649 TI - Legislative session: the year of health care reform. PMID- 18512650 TI - In your hands: a case for hand hygiene. PMID- 18512651 TI - Iowa Healthcare Collaborative equips physicians for change. PMID- 18512652 TI - The next generation of physicians: what will they need? PMID- 18512653 TI - Research benefits for hypothetical HIV vaccine trials: The views of Ugandans in the Rakai District. PMID- 18512654 TI - Effective use of consent forms and interactive questions in the consent process. PMID- 18512655 TI - Clinical research in low-literacy populations: using teach-back to assess comprehension of informed consent and privacy information. PMID- 18512656 TI - Orthodontics in a quantum world I: the rationale for a new approach. AB - Advances in physics and cell biology are changing how science views studies about the body. The first part of the article is an, overview of these advances. The second part is a working hypothesis as to how these changes could affect orthodontic diagnosis and treatment. An example is given of how this thinking might apply. PMID- 18512657 TI - Three dimensional computed technology--a new standard of care. AB - I-Cat Scan Cone Beam CT (CBCT) technology is rapidly replacing two dimensional radiography in the medical world. Virtual patients may be viewed on a screen and multiple sectional views observed in order to obtain more definitive diagnostic and treatment options. The authors give a brief overview of some opportunities available using I-Cat technology as well as multiple actual cases where subjective diagnosis has been reduced substantially through CBCT technology. PMID- 18512658 TI - Class III management part II: a clinical case. PMID- 18512659 TI - Treatment timing. PMID- 18512660 TI - Case finishing part 1: it starts at the beginning. PMID- 18512661 TI - Continuing the journey to success. PMID- 18512662 TI - The cheerleader who won't shake her pom pons. PMID- 18512663 TI - Cost benefit considerations of preventing elderly falls through environmental modifications to homes in Hana, Maui. AB - The community of Hana, Hawai'i began a program of home modifications to help their elderly prevent falls. We estimated the cost benefit of these modifications from construction costs and published reports of effectiveness and cost of treating falls. We interviewed clients to determine risk of falling. The average cost of home modifications was $800. The average annual averted medical cost of falling was $1728. PMID- 18512664 TI - Suicidal behavior trends in a pediatric population in Hawai'i. AB - In this retrospective 2-year study the clinical and demographic characteristics of pediatric patients in Hawai'i evaluated for suicide attempts (or possible suicide attempts) at a local children's hospital were examined. Our intentions were to uncover any trends in this unique, culturally diverse population that has not been studied extensively. Sixty-five patients were identified, of which 86% were girls with the highest age demographic at 15-16 years of age (34%) and medication overdose the most common mechanism. Ethnicity trends could not reliably be ascertained. The circumstances surrounding the suicidal behavior; adjustment disorder being a common diagnosis; and approximately half of the population not intending to complete suicide suggests possible impulsivity. Thus, the results have the potential to direct interventions for community suicide prevention programs in Hawai'i, with a focus on coping strategies and medication overdose education. PMID- 18512665 TI - Prime-time television exposure to high priority school-aged social-developmental issues. AB - BACKGROUND: The purpose of this study is to quantify the material children and adolescents are exposed to while watching prime-time television so that school educators, health professionals, and parents can focus on issues of maximum exposure that must be addressed. METHODS: Prime-time programming was recorded from 7 p.m. to 10 p.m. Hawaiian Standard Time daily for 2 weeks in July 2005. Recordings were then viewed to identify social behaviors of interest. RESULTS: Each hour on average, sex was referenced 1.8 times, drugs 0.6 times, tobacco 0.3 times, alcohol 2.4 times, and violence/crime 6.0 times per network. Messages advocating exercise, anti-drug advocacy, and anti-smoking advocacy were each shown 0.2 times per hour; while anti-alcohol advocacy was shown 0.1 times per hour. CONCLUSION: School educators, health professionals, and parents must recognize that prime-time television frequently exposes viewers to issues that are of critical importance to the health and social development of school-aged children and adolescents. PMID- 18512666 TI - Assessment of disaster training needs for physicians and other health professionals serving vulnerable populations in Hawai'i. PMID- 18512667 TI - Providing psychosocial support to men with prostate cancer and their wives. PMID- 18512668 TI - Issues in medical malpractice XXI. PMID- 18512669 TI - Glycosylation analysis of glycoproteins and proteoglycans using capillary electrophoresis-mass spectrometry strategies. AB - This review highlights recent developments in glycosylation analysis by modern MS in combination with CE based preseparation. Focused on CE-MS strategies aimed for glycotyping, the review addresses the detailed glycoform analysis of glycoproteins, glycopeptides, and proteoglycans. Glycoform analysis in the context of modern glycoproteomics is briefly addressed, as well. CZE, CIEF, and frontal analysis CE approaches hyphenated to high-resolution multistage MS for the detailed analysis of protein-linked glycan structures are overviewed in a comprehensive way. Advantages and limitations of various methodological approaches and techniques as well as mass spectrometric instrumentation are discussed in the particular context of glycoanalysis. PMID- 18512673 TI - Capillary electrophoresis hyphenated to inductively coupled plasma-mass spectrometry: a novel approach for the analysis of anticancer metallodrugs in human serum and plasma. AB - The development of metal-based chemotherapeutics lacks methods which are capable of providing early indication on the potential of new metal complexes as future anticancer drugs. Since most of these compounds are administered intravenously, serum proteins are the first available biological binding partners in the bloodstream. For platinum-based anticancer drugs the interaction with serum proteins is regarded as an important contribution to the side effects accompanying chemotherapy. In contrast, newly developed ruthenium compounds are thought to be transported into the tumor in a protein-bound form. In here, the application of CE hyphenated to inductively coupled plasma (ICP)-MS, applying Polybrene-coated capillaries, is demonstrated for studying the interaction of indazolium [trans-tetrachlorobis(1H-indazole)ruthenate(III)] (KP1019) with HSA and transferrin, which are important transport proteins. Furthermore, the applicability of the method to human serum and plasma and, more importantly, to real-world patient samples was proven. KP1019 was found to bind to a high degree to HSA both in serum, plasma and the patient samples. Only minor fractions of ruthenium were found attached to other proteins. PMID- 18512674 TI - A microfluidic study of mechanisms in the electrophoresis of supercoiled DNA. AB - In this work, microfluidic chips were used to study the electrophoresis of supercoiled DNA (SC DNA) in agarose. This system allowed us to study the electrophoretic and trapping behaviours of SC DNA of various lengths, at various fields and separation distances. Near a critical electric field the DNA is trapped such that the concentration falls exponentially with distance. The trapping of such circular DNA has been explained in terms of the 'lobster trap' or 'impalement' model where shorter fibres become trapping sites at higher fields, leading to an ongoing (and gradual) increase in trapping with increasing field. By contrast, the present study suggests that under some circumstances the traps have a barrier such that only when the DNA has sufficient energy (at high enough fields) can it become trapped, leading to a sudden transition in behaviours at the critical field. We propose an 'activated impalement' mechanism of trapping in which only at sufficiently high fields is the SC DNA impaled and trapped for long times. The critical electric field appears to be inversely proportional to the length of the DNA molecule, suggesting that the force required to impale the SC DNA is constant. PMID- 18512675 TI - Capillary electrophoresis of acidic oligosaccharides from human milk. AB - Interest in defining the array of oligosaccharides of human milk has been increasing. Pathogens that bind glycans on their host mucosal surfaces may be inhibited by human milk oligosaccharides. It has been postulated that acidic oligosaccharides in human milk may inhibit binding by pathogens that bind acidic glycans in the gut, but testing this hypothesis requires their reliable quantification in milk. Sialyloligosaccharides of human milk have been quantified by HPLC and CE. A recent CE technique uses the MEKC mode with direct detection at 205 nm to resolve and quantify, in the native form, the 12 most dominant sialyloligosaccharides of human milk in a single 35-min run. The method gives a linear response from 39 to 2500 microg/mL with a coefficient of variation between 2 to 9% and accuracy from 93 to 109%. This was used to detect variation in expression of specific sialyloligosaccharides in milk. Individual sialyloligosaccharide concentrations in milk differ among individual donors and between less and more mature milk. Thus, CE can be used to measure variation in sialyloligosaccharide expression in milk, and thereby test the relationship of this variation-to-variation in risk of specific diseases in breastfed infants. PMID- 18512676 TI - Validation of capillary zone electrophoresis and capillary isoelectric focusing separations optimized for the characterization of two recombinant products of the birch pollen allergen Bet v 1a. AB - CZE and CIEF separation systems, both developed previously for a quality control of two recombinant products of the major birch pollen allergen Bet v 1a of Betula verrucosa, were validated including aspects of the International Conference on Harmonization. One product contained carbamylated variants as impurities. Linearity of response was confirmed by Mandel's fitting test between 0.028 and 1.90 mg/mL for CZE and between 0.016 and 0.26 mg/mL for CIEF. Repeatability and intermediate precision were evaluated for the effective mobility (mu(eff)) in CZE, for relative mobilization time in CIEF and the peak area ratio of Bet v 1a. LOQ for Bet v 1a was between 10 and 23 microg/mL for both methods. Evaluation of robustness for CZE revealed susceptibility of micro(eff) of Bet v 1a to alterations in of buffer pH and separation temperature. Selectivity was impaired by an increase in temperature, pH, and buffer concentration. In addition, pH variations influenced the separation profile of impurities. For CIEF, the ratio of narrow pH range carrier ampholytes is the critical parameter to retain robustness. Results demonstrate the suitability of both separation systems to discriminate between nonmodified Bet v 1a and carbamylated variants in the selected recombinant allergen products. PMID- 18512677 TI - Capillary electrophoresis with inductively coupled plasma-mass spectrometric and electrospray time of flight mass spectrometric detection for the determination of arsenic species in fish samples. AB - CE was coupled to inductively coupled plasma MS (ICP-MS) and ESI-MS to identify and quantify the arsenic species arsenobetaine (AsB), arsenite (As(III)), arsenate (As(V)), and dimethylarsinic acid (DMA). A GC-flame ionization detector (FID)-based German standard method and ICP-MS were used for validation of the data obtained for arsenobetaine and total arsenic, respectively. LODs obtained with the CE-ESI-TOF-MS method were 1.0x10(-7) M for AsB, 5.0x10(-7) M for DMA, and 1.0x10(-6) M for As(III) and As(V). For the CE-ICP-MS method, LODs were 8.5x10(-8) M for AsB, 9.5x10(-8) M for DMA, 9.3x10(-8) M for As(III), and 6.2x10( 8) M for As(V). While CE-ICP-MS provided high sensitivity and better reproducibility for quantitative measurements, CE-ESI-MS with a TOF mass analyzer proved to be valuable for species identification. With this setup, fish samples were prepared and analyzed and the obtained data were successfully validated with the independent methods. PMID- 18512679 TI - Biomarker discovery in psychiatric disorders. AB - Schizophrenia is a multifaceted neuropsychiatric disorder. Its onset is the result of complex interactions between genetic, developmental and environmental factors. It almost certainly presents a heterogeneous group of aetiologies which may not be reflected in the symptomatic/clinical presentation of patients. Therefore, a better molecular understanding of the disease onset and progression is urgently needed. The high complexity of the disorder and the heterogeneity of patient populations account for the slow progress of biomarker discovery approaches. Multi-omics profiling approaches can be employed to investigate large numbers of patient and control samples in a single experiment. These large scale experiments are required to identify disease intrinsic molecular signatures as well as patient subgroups with potentially distinct biochemical pathways underpinning their symptoms. In this overview, we describe some of the most important challenges for biomarker discovery for psychiatric disorders and emphasize how these problems contribute to the requirement of large sample numbers. Results of MS-based protein profiling studies in schizophrenia research are reviewed and technical advantages and difficulties of the methodologies described. We outline recent technological advances that generated impressive results in other areas of research and point to their applicability for biomarker discovery in psychiatric disorders. PMID- 18512680 TI - Photopatterning of tough single-walled carbon nanotube composites in microfluidic channels and their application in gel-free separations. AB - We report on the photopatterning of single carbon nanotube composites with soft hydrogel polymers in glass microchannels. Since the hydrogels by themselves are able to withstand liquid flow within the microchannels, we covalently combined them with single-walled carbon nanotubes to impart mechanical strength. We attempted this approach by patterning the gels within the microchannels without prior surface modifications. Our results show that the 1-cm nanocomposite hydrogels are far stronger than the free hydrogels. Moreover, the nanocomposites were able to concentrate and separate proteins within a 1.5-cm distance using gel free buffers. The separation cannot only be tuned by changing the running buffer; the lack of gels in the running buffer reduces the chance of channel blockage and thus the lifetime of the device is prolonged. The usefulness of the patterned nanocomposites may be extended by a wide selection of nanocomposite properties and monomers to find a broad range of applications in lab-on-chip technology. PMID- 18512681 TI - Multiplexed quantitation of endogenous estrogens and estrogen metabolites in human peritoneal fluid. AB - Endogenous estrogens and estrogen metabolites (EM) in human peritoneal fluid may play an important role in health and disease, yet little is known regarding their types and levels present in human peritoneal fluid, primarily due to the lack of an analytical method that is capable of directly quantifying their absolute abundances. In this report, we describe the application of a capillary LC-MS/MS method for identifying and quantifying biologically active and total endogenous EM in human peritoneal fluid. The method requires only 50 muL of peritoneal fluid, yet can quantify 13 distinct EM. Calibration curves for each EM were linear over a 10(3)-fold concentration range and the lower LOQ was 50 fg on column. For a charcoal stripped human peritoneal fluid sample containing 10 pg/mL of each EM, accuracy ranged from 83 to 118%, and intrabatch precision ranged from 0.2 to 4.4% RSD and interbatch precision ranged from 5.5 to 15.5% RSD. The analyses of human female peritoneal fluid shows that at least 10 biologically active and 11 total endogenous EM can be positively identified and quantitatively measured. Many of the biologically active forms are present in high abundance and possess distinct biological activities which warrant further study. Although micellar EKC gave baseline separation of a standard mixture of 10 EM, the LOQs using UV detection were not suitable for the assay of the low level estrogens in biological samples. PMID- 18512682 TI - Implementation of a protein profiling platform developed as an academic pharmaceutical industry collaborative effort. AB - As much attention has devoted to the proteome research during the last few years, biomarker discovery has become an increasingly hot area, potentially enabling the development of new assays for diagnosis and prognosis of severe diseases. This is the field of research interest where efforts originating from both academic and industrial groups should jointly work on solutions. In this paper, we would like to demonstrate the fruitful combination of both research domains where the scientific crossroads sprout fresh ideas from the basic research domain and how these are refined and tethered to industrial standards. We will present an approach that is based on novel microfluidic devices, utilizing their benefits in processing small-volume samples. Our biomarker discovery strategy, built around this platform, involves optimized samples processing (based on SPE and sample enrichment) and fast MALDI-MS readout. The identification of novel biomarkers at low-abundance level has been achieved by the utilization of a miniaturized sample handling platform, which offers clean-up and enrichment of proteins in one step. Complete automation has been realized in the form of a unique robotic instrumentation that is able to extract and transfer 96 samples onto standard MALDI target plates with high throughput. The developed platform was operated with a 60 sample turnaround per hour allowing sensitivities in femtomol regions of medium- and low-abundant target proteins from clinical studies on samples of multiple sclerosis and gastroesophageal reflux disease. Several proteins have been identified as new biomarkers from cerebrospinal fluid and esophagus epithelial cells. PMID- 18512683 TI - Analysis of anthocyanins in red onion using capillary electrophoresis-time of flight-mass spectrometry. AB - For the first time, a capillary electrophoresis-time of flight-mass spectrometry analysis method for detecting anthocyanins in red onion was developed. The analysis method included the use of silica capillaries coated with poly-LA 313 (polycationic amine-containing polymer) and an MS-compatible volatile background electrolyte (BGE). The method was environmentally friendly and sensitive; and its rapidness combined with an acidic BGE helped in preventing anthocyanin degradation. By using high-resolution TOF-MS with pre-run tuning of masses, low mass errors were achieved in the determination of conjugated anthocyanins in red onion, and a simultaneous up-front fragmentation provided confirmation of the aglycon backbone for their secure identification. Most anthocyanins (at least seven out of ten) known in red onion from the literature were found, as well as one new for this matrix. PMID- 18512684 TI - Immunohistochemical identification of the extravillous trophoblast during the placentation of the degu (Octodon degus). AB - Recent data indicate that placentation in Octodon degus is similar to that in humans, making it a potential animal model for studies in human placental pathologies related to alterations in the migration of the extravillous trophoblast (EVT). Our objective was to immunohistochemically identify degu EVT during placentation by using cytoskeletal protein markers to establish the normal migratory pattern of the EVT. Fifteen O.degus were divided into three equal groups: day 27, 60, and 84 of gestation. The placentas were immunostained for cytokeratin (CK) and alpha smooth muscle actin (SMA). At day 27, the migrating EVT immunostained for SMA but not for CK. Once the EVT was incorporated in the maternal vessels (day 60) it was positive for CK but negative for SMA. The smooth muscle cells of the mesometrial arteries that remained after EVT invasion were positive for SMA. At day 84, the media muscular layer had partially regenerated but some EVT was still present. Furthermore, at day 27 cyclooxygenase-1 (COX-1) was detected in the endothelium of the maternal decidual vessels. Our results suggest that during the early stages of placentation, the cytoskeletal organization of the actin network of the migrating EVT corresponds to that of a cell with motile behavior. Once the EVT invaded the spiral arteries, the cytoskeleton reorganized, adopting the structure of an epithelial-like cell, expressing CK intermediate filaments. The media muscle layer regenerated near the end of gestation but some EVT remained. During EVT formation the endothelium of the maternal decidual vessels immunostained for COX-1. PMID- 18512685 TI - Antidepressant drug-induced stimulation of mouse hippocampal neurogenesis is age dependent and altered by early life stress. AB - The continuous generation of new neurons in the adult hippocampus exhibits remarkable plasticity. Decreased neurogenesis is thought to underlie depression like behaviors, and increased neurogenesis is thought to occur following antidepressant drug treatment. Studies on different strains of mice, however, yielded contrasting results with regard to the link between behavioral modifications induced by antidepressant drugs or environmental enrichment and changes in adult hippocampal neurogenesis. Therefore, we conducted a comparative study on the inbred strains Balb/c and C57Bl/6 that differ substantially in emotionality, stress reactivity, and behavioral responses to chronic antidepressant drugs. Quantitative assessments of progenitor cell proliferation and immature neuronal differentiation in the dentate gyrus revealed that, despite significantly different basal proliferation rates between both strains, neither strain exhibited changes in adult neurogenesis after exposure to early life stress or adult chronic fluoxetine treatment. A stimulatory effect of fluoxetine on adult hippocampal neurogenesis was only detected when treatment was initiated during adolescence, and this effect was abolished in mice exposed to early life stress, a prominent risk factor for developing adult-onset depression-like behaviors. Thus, in both strains of mice neither adult fluoxetine treatment nor adolescent fluoxetine treatment following early life stress exposure increased the proliferation and early differentiation of adult neural progenitor cells. PMID- 18512686 TI - Relatedness in wild chimpanzees: influence of paternity, male philopatry, and demographic factors. AB - In chimpanzees (Pan troglodytes), high-ranking males are expected to have high reproductive success and females typically emigrate upon reaching maturity. Although high average relatedness among males in the same social groups has been assumed, previous reports have indicated that relatedness among males is not necessarily significantly higher than that among females. The paternity of 11 offspring and the relatedness of 50 individuals in the M group of chimpanzees at Mahale Mountains National Park, Tanzania, were investigated using DNA analyses. We determined the fathers of 10 offspring. Two different alpha males sired a total of five offspring, whereas the other males had low reproductive success. The proportion of paternal half-sibling pairs among the 10 offspring was 15.6%. The average relatedness among mature males was significantly higher than that among mature females. The existence of an old male and the long tenure of one alpha male may have contributed to this significant difference. The average dyadic relatedness among mature natal individuals was significantly higher than that in natal-immigrant pairs in which the individuals came from different groups. The average relatedness among immigrant females was similar to that in pairs of natal and immigrant females, suggesting that the immigrants came from various groups. Thus, female transfer acts to maintain low average relatedness within the group. A comparison of our results to those from other study sites suggests that although the average relatedness among adult males does not reach the level of half-siblings, under some circumstances it can exceed the relatedness of females. PMID- 18512687 TI - The genetic divergence of prehistoric populations of the south-central Andes as established by means of craniometric traits. AB - The peopling of the south-central Andean region can be determined by exploring a combination of cultural, economic, and biological factors that influence the structure of populations and determine particular dispersals of gene frequencies. Quantitative characters from 1,586 adult crania of both sexes from northern Chile, northwestern Argentina, and the Cochabamba valleys in Bolivia were analyzed employing multivariate statistical analyses. Biological distances, representing phenotypic variation between these regions and their subregions, were studied within a population genetics framework. An analysis of Mahalanobis D(2) distances establishes two principle directions of interaction: the first between the Cochabamba valleys and northern Chile, and the second between the Cochabamba region and northwestern Argentina. The Chile and Argentina regions are shown to be less related to each other than each is to the Bolivian region. A higher mean genetic divergence is found for the entire region (F(ST) = 0.195); with northwestern Argentina having the highest spatial isolation (F(ST) = 0.143) and northern Chile the lowest (F(ST) = 0.061). These results allow us to propose a populating model based on the dispersion of several lines from a common ancestral population similar to those who inhabited the Cochabamba valleys. These lines differentiated themselves in time and space according to the effective size and the rate of gene flow, eventually producing the human groups which inhabited the valleys of northern Chile and northwestern Argentina. PMID- 18512688 TI - Supramolecular synthons on surfaces: controlling dimensionality and periodicity of tetraarylporphyrin assemblies by the interplay of cyano and alkoxy substituents. AB - The self-assembly of three porphyrin derivatives was studied in detail on a Cu(111) substrate by means of scanning tunneling microscopy (STM). All derivatives have two 4-cyanophenyl substituents in diagonally opposed meso positions of the porphyrin core, but differ in the nature of the other two meso alkoxyphenyl substituents. At coverages below 0.8 monolayers, two derivatives form molecular chains, which evolve into nanoporous networks at higher coverages. The third derivative self-assembles directly into a nanoporous network without showing a one-dimensional phase. The pore-to-pore distances for the three networks depend on the size and shape of the alkoxy substituents. All observed effects are explained by 1) different steric demands of the alkoxy residues, 2) apolar (mainly dispersion) interactions between the alkoxy chains, 3) polar bonding involving both cyanophenyl and alkoxyphenyl substituents, and 4) the entropy/enthalpy balance of the network formation. PMID- 18512689 TI - Reductive degradation of nido-1-CB8H12 into smaller-cage carborane systems via new monocarbaboranes [arachno-5-CB8H13](-) and closo-2-CB6H8. AB - Treatment of the nido-1-CB8H12 (1) carborane with NaBH4 in THF at ambient temperature led to the isolation of the stable [arachno-5-CB8H13]- (2(-)), which was isolated as Na+[5-CB8H13]-.1.5 THF and PPh4 +[5-CB8H13]- in almost quantitative yield. Compound 2(-) underwent a boron-degradation reaction with concentrated hydrochloric acid to afford the arachno-4-CB7H13 (3) carborane in 70 % yield, whereas reaction between 2(-) and excess phenyl acetylene in refluxing THF gave the [closo-2-CB6H7]- (4-) in 66 % yield. Protonation of the Cs+4(-) salt with concentrated H2SO4 or CF3COOH in CH2Cl2 afforded a new, highly volatile 2 CB6H8 (4) carborane in 95 % yield, the deprotonation of which with Et3N in CH2Cl2 leads quantitatively to Et3NH+[2-CB6H7](-) (Et3NH+4(-)). Both compounds 4- and 4 can be deboronated through treatment with concentrated hydrochloric acid in CH2Cl2 to yield the carbahexaborane nido-2-CB5H9 (5) in 60 % yield. New compounds 2-, 3, and 4 were structurally characterised by the ab initio/GIAO/MP2/NMR method. The method gave superior results to those carried out using GIAO-HF when relating the calculated 11B NMR chemical shifts to experimental data. PMID- 18512690 TI - A molecular orbital rationalization of ligand effects in N2 activation. AB - Molecular orbital theory has been used to study a series of [(micro-N2){ML3}2] complexes as models for dinitrogen activation, with M=Mo, Ta, W, Re and L=NH2, PH2, AsH2, SbH2 and N(BH2)2. The main aims of this study have been to provide a thorough electronic analysis of the complexes and to extend previous work involving molecular orbital analyses. Molecular orbital diagrams have been used to rationalize why for L=NH2 ligand rotation is important for the singlet state but not the triplet, to confirm the effect of ligand pi donation, and to rationalize the importance of the metal d-electron configuration. The outcomes of this study will assist with a more in-depth understanding of the electronic basis for N2 activation and allow clearer predictions to be made about the structure and multiplicity of systems involved in transition-metal catalysis. PMID- 18512691 TI - Reactions of nitroheteroarenes with carbanions: bridging aromatic, heteroaromatic, and vinylic electrophilicity. AB - The relative rate constants for the vicarious nucleophilic substitution (VNS) of the anion of chloromethyl phenyl sulfone (1-) with a variety of nitroheteroarenes, for example, nitropyridines, nitropyrroles, nitroimidazoles, 2 nitrothiophene, and 4-nitropyrazole, have been determined by competition experiments. It was shown that nitropyridines are approximately four orders of magnitude more reactive than nitrobenzene. Among the five-membered heterocycles 2 nitrothiophene is the most active followed by nitroimidazoles and 4 nitropyrazole. Nitropyrroles are the least electrophilic nitroheteroarenes with reactivities comparable to nitrobenzene. Quantum chemically calculated methyl anion affinities (B3LYP/6-311G(d,p)//B3LYP/6-31G(d)) of the nitroarenes correlated only moderately with the partial relative rate constants. The correlation of these activities with the LUMO energies of nitroarenes is even worse. By measuring the second-order rate constants of the addition of 1- to nitroarenes and to diethyl arylidenemalonates 10, it was possible to link the electrophilic reactivities of nitroheteroarenes with the comprehensive electrophilicity scale based on the linear-free-energy-relationship log k(20 degrees C)=s(N+E). PMID- 18512693 TI - The Influence of a gradient static magnetic field on an unstirred Belousov Zhabotinsky reaction. AB - It is believed that static magnetic fields (SMF) cannot affect the pattern formation of the Belousov-Zhabotinsky (BZ) reaction, which has been frequently studied as a simplified experimental model of a nonequilibrium open system, because SMF produces no induced current and the magnetic force of SMF far below 1 T is too low to expect the effects on electrons in the BZ reaction. In the present study, we examined whether the velocity of chemical waves in the unstirred BZ reaction can be affected by a moderate-intensity SMF exposure depending on the spatial magnetic gradient. The SMF was generated by a parallel pair of attracting rectangular NdFeB magnets positioned opposite each other. The respective maximum values of magnetic flux density (B(max)), magnetic flux gradient (G(max)), and the magnetic force product of the magnetic flux density its gradient (a magnetic force parameter) were 206 mT, 37 mT/mm, and 3,000 mT(2)/mm. The ferroin-catalyzed BZ medium was exposed to the SMF for up to 16 min at 25 degrees C. The experiments demonstrated that the wave velocity was significantly accelerated primarily by the magnetic gradient. The propagation of the fastest wave front indicated a sigmoid increase along the peak magnetic gradient line, but not along the peak magnetic force product line. The underlying mechanisms of the SMF effects on the anomalous wave propagation could be attributed primarily to the increased concentration gradient of the paramagnetic iron ion complexes at the chemical wave fronts induced by the magnetic gradient. PMID- 18512692 TI - Plasticity of lumbosacral propriospinal neurons is associated with the development of autonomic dysreflexia after thoracic spinal cord transection. AB - Complete thoracic (T) spinal cord injury (SCI) above the T6 level typically results in autonomic dysreflexia, an abnormal hypertensive condition commonly triggered by nociceptive stimuli below the level of SCI. Overexpression of nerve growth factor in the lumbosacral spinal cord induces profuse sprouting of nociceptive pelvic visceral afferent fibers that correlates with increased hypertension in response to noxious colorectal distension. After complete T4 SCI, we evaluated the plasticity of propriospinal neurons conveying visceral input rostrally to thoracic sympathetic preganglionic neurons. The anterograde tracer biotinylated dextran amine (BDA) was injected into the lumbosacral dorsal gray commissure (DGC) of injured/nontransected rats immediately after injury (acute) or 2 weeks later (delayed). At 1 or 2 weeks after delayed or acute injections, respectively, a higher density (P < 0.05) of BDA(+) fibers was found in thoracic dorsal gray matter of injured vs. nontransected spinal cords. For corroboration, fast blue (FB) or cholera toxin subunit beta (CTb) was injected into the T9 dorsal horns 2 weeks postinjury/nontransection. After 1 week transport, more retrogradely labeled (P < 0.05) DGC propriospinal neurons (T13-S1) were quantified in injured vs. nontransected cords. We also monitored immediate early gene c-fos expression following colorectal distension and found increased (P < 0.01) c-Fos(+) cell numbers throughout the DGC after injury. Collectively, these results imply that, in conjunction with local primary afferent fiber plasticity, injury-induced sprouting of DGC neurons may be a key constituent in relaying visceral sensory input to sympathetic preganglionic neurons that elicit autonomic dysreflexia after high thoracic SCI. PMID- 18512694 TI - Extremely low frequency electromagnetic fields (ELF-EMFs) induce in vitro angiogenesis process in human endothelial cells. AB - Effects of extremely low frequency (ELF) electromagnetic fields (EMFs) on activation of angiogenesis were analysed using cultured umbilical human vein endothelial cells (HUVECs). The cultures were exposed to a sinusoidal EMF to intensity of 1 mT, 50 Hz for up to 12 h. EMFs increased the degree of endothelial cell proliferation and tubule formation, coupled by an acceleration in the process of wound healing. Since this process is physiologically accompanied by a large modification in the structural organization of actin and focal adhesions, we analyzed the rearrangement of some cytoskeleton elements demonstrating a major reorganization of the fibres and of the focal adhesion complexes after EMF exposure. Finally, Western blot analysis revealed a significant increase in phosphorylation as well as the overall expression of VEGF receptor 2 (KDR/Flk-1) suggesting that EMFs may modulate in vitro some endothelial functions correlated to angiogenesis through signal transduction pathways dependent on VEGF. PMID- 18512695 TI - Spindle disturbances in human-hamster hybrid (AL) cells induced by mobile communication frequency range signals. AB - The production of spindle disturbances in FC2 cells, a human-hamster hybrid (A(L)) cell line, by non-ionizing radiation was studied using an electromagnetic field with a field strength of 90 V/m at a frequency of 835 MHz. Due to the given experimental conditions slide flask cultures were exposed at room temperature in a microTEM (transversal electromagnetic field) cell, which allows optimal experimental conditions for small samples of biological material. Numerical calculations suggest that specific absorption rates of up to 60 mW/kg are reached for maximum field exposure. All exposure field parameters--either measured or calculable--are precisely defined and, for the first time, traceable to the standards of the SI system of physical units. Compared with co-incident negative controls, the results of two independently performed experiments suggest that exposure periods of time from 0.5 to 2 h with an electric field strength of 90 V/m are spindle acting agents as predominately indicated by the appearance of spindle disturbances at the ana- and telophase stages (especially lagging and non disjunction of single chromosomes) of cell divisions. The spindle disturbances do not change the fraction of mitotic cells with increasing exposure time up to 2 h. Due to the applied experimental conditions an influence of temperature as a confounder parameter for spindle disturbances can be excluded. PMID- 18512696 TI - Acute radio frequency irradiation does not affect cell cycle, cellular migration, and invasion. AB - Although in vitro studies have been previously conducted to determine the biological effects of radio frequency (RF) radiation, it has not yet been determined whether or not RF radiation poses a potential hazard. This study was conducted to determine whether RF radiation exposure exerts detectable effects on cell cycle distribution, cellular invasion, and migration. NIH3T3 mouse fibroblasts were exposed to 849 MHz of RF radiation at average SAR values of 2 or 10 W/kg for either 1 h, or for 1 h per day for 3 days. During the exposure period, the temperature in the exposure chamber was maintained isothermally by circulating water throughout the cavity. Cell cycle distribution was analyzed at 24 and 48 h after exposure, by flow cytometry. We detected no statistically significant differences between the sham-exposed and RF radiation-exposed cells. Cellular invasion and migration were assessed by in vitro Matrigel invasion and Transwell migration assays. The RF radiation-exposed groups evidenced no significant changes in motility and invasiveness compared to the sham-exposed group. However, the ionizing radiation-exposed cells, used as a positive control group, manifested dramatic alterations in their cell cycle distribution, cellular invasiveness, and migration characteristics. Our results show that 849 MHz RF radiation exposure exerts no detectable effects on cell cycle distribution, cellular migration, or invasion at average SAR values of 2 or 10 W/kg. PMID- 18512697 TI - Exposure of seeds to static magnetic field enhances germination and early growth characteristics in chickpea (Cicer arietinum L.). AB - Seeds of chickpea (Cicer arietinum L.) were exposed in batches to static magnetic fields of strength from 0 to 250 mT in steps of 50 mT for 1-4 h in steps of 1 h for all fields. Results showed that magnetic field application enhanced seed performance in terms of laboratory germination, speed of germination, seedling length and seedling dry weight significantly compared to unexposed control. However, the response varied with field strength and duration of exposure without any particular trend. Among the various combinations of field strength and duration, 50 mT for 2 h, 100 mT for 1 h and 150 mT for 2 h exposures gave best results. Exposure of seeds to these three magnetic fields improved seed coat membrane integrity as it reduced the electrical conductivity of seed leachate. In soil, seeds exposed to these three treatments produced significantly increased seedling dry weights of 1-month-old plants. The root characteristics of the plants showed dramatic increase in root length, root surface area and root volume. The improved functional root parameters suggest that magnetically treated chickpea seeds may perform better under rainfed (un-irrigated) conditions where there is a restrictive soil moisture regime. PMID- 18512698 TI - Skull shape and feeding strategy in Sphenodon and other Rhynchocephalia (Diapsida: Lepidosauria). AB - The Rhynchocephalia are a group of small diapsid reptiles that were globally distributed during the early Mesozoic. By contrast, the only extant representatives, Sphenodon punctatus and S. guntheri (Tuatara), are restricted to New Zealand off-shore islands. The Rhynchocephalia are widely considered to be morphologically uniform but research over the past 30 years has revealed unexpected phenotypic and taxonomic diversity. Phylogenetically "basal taxa" generally possess relatively simple conical or columnar teeth whereas more derived taxa possessed stouter flanged teeth and sophisticated shearing mechanisms: orthal in some (e.g., Clevosaurus hudsoni) and propalinal in others (e.g., S. punctatus). This variation in feeding apparatus suggests a wide range of feeding niches were exploited by rhynchocephalians. The relationship of skull shape to skull length, phylogenetic grouping, habit, and characters relating to the feeding apparatus are explored here with geometric morphometric analysis on two-dimensional landmarks. Principle components analysis demonstrates that there are significant differences between phylogenetic groups. In particular, Sphenodon differs significantly from all well known fossil taxa including the most phylogenetically basal forms. Therefore, it is not justifiable to use Sphenodon as a solitary outgroup when studying skull shape and feeding strategy in squamates; rhynchocephalian fossil taxa also need to be considered. There are also significant differences between the skull shapes of aquatic taxa and those of terrestrial taxa. Of the observed variation in skull shape, most variation is subsumed by variation in dentary tooth base shape, the type of jaw movement employed (e.g., orthal vs. propalinal) and the number of palatal tooth rows. By comparison, the presence or absence of flanges, dentary tooth number and palatal tooth row orientation subsume much less. Skull length was also found to be a poor descriptor of overall skull shape. Compared to basal rhynchocephalians members of more derived terrestrial radiations possess an enlarged postorbital area, a high parietal, and a jaw joint positioned ventral to the tooth row. Modification of these features is closely associated with increased biting performance and thus access to novel food items. Some of these same trends are apparent during Sphenodon ontogeny where skull growth is allometric and there is evidence for ontogenetic variation in diet. PMID- 18512699 TI - Natural and anthropogenic determinants of genetic structure in the largest remaining population of the endangered golden-brown mouse lemur, Microcebus ravelobensis. AB - Genetic differentiation between natural populations is best understood as a result of both natural and anthropogenic factors. Genetic studies on large populations still living under relatively undisturbed conditions are extremely valuable to disentangle these influences. The effect of three natural (geographic distance, landscape, dispersal) factors and two anthropogenic factors (road, savannah) on gene flow was analyzed in the largest remaining forest region in the range of the endangered golden-brown mouse lemur in Madagascar. A total of 187 individuals from 12 sites were sampled and genotyped at eight polymorphic microsatellite loci. All sites exhibited similar levels of genetic variation. The level of genetic differentiation was low to moderate with pairwise F(ST) values ranging from -0.002 to 0.12, but most were significant and all sites exhibited high self-assignment rates. A spatial autocorrelation analysis was performed at two geographic scales revealing a pattern of isolation-by-distance and suggesting that no clear differences exist between male and female local dispersal. Two Bayesian approaches revealed that a stretch of savannah represented a significant barrier to movement, whereas the influence of the road on gene flow was less clear. Finally, we found that landscape characteristics, in particular altitude, play a role in the functional connectivity of the sites. The study underlines the importance of studies in relatively undisturbed conditions for the interpretation of population genetics data in fragmented environments. The results are discussed in terms of their conservation relevance for forest-dwelling animals such as most primate species. PMID- 18512700 TI - Cathodoluminescence study of semiconductor oxide micro- and nanostructures grown by vapor deposition. AB - Nano- and microstructures of SnO(2), In(2)O(3) and ZnO have been grown during thermal treatment of compacted powders under argon flow. Indium-doped SnO(2) tube shaped structures with rectangular cross-section are obtained by adding a fraction of In(2)O(3) to the starting SnO(2) powder. In-rich nanoislands were found to grow on some edges of the tubes. ZnO nanostructures doped with Sn or Eu were grown by adding SnO(2) and Eu(2)O(3) powder, respectively, to the ZnO precursor powder. All the samples have been characterized by the emissive and cathodoluminescence (CL) modes of scanning electron microscopy. CL images from SnO(2):In and In(2)O(3):Sn tubes and islands show a higher emission from the Sn rich structures related to oxygen deficiency. CL of doped ZnO enables to detect the presence of dopant in specific regions or structures. CL appears to be a useful technique to study optical and electronic properties of semiconductor oxide nanostructures. PMID- 18512701 TI - Evaluation of surface roughness and nanostructure of indium tin oxide (ITO) films by atomic force microscopy. AB - Indium tin oxide was deposited on a glass (soda lime glass) by radiofrequency sputtering system at different sputtering gas (argon/oxygen 90/10%) pressures (20 34 mTorr) at room temperature. The sputtering rate was affected by the sputtering gas pressure. The optimum sputtering gas pressure was found to be 27 mTorr. The samples at different thicknesses (168, 300, 400, 425, 475, 500 and 630 nm) were deposited on the substrate. Transparency, electrical conductivity and surface roughness of the films were characterized. The samples were annealed at 350, 400 and 450 degrees C to evaluate annealing process effects on the concerned parameters and, therefore, the above-mentioned measurements were repeated again. The films exhibited reasonable optical transmittance and electrical conductivity and greatly improved after annealing. The characterization was focused on the scanning of the film surfaces before and after annealing, which has a prominent effect on the optical properties of the films. Film surfaces were scanned by scanning probe microscopy in contact atomic force mode. The most consideration was devoted to image analysis. PMID- 18512702 TI - Glial architecture of the ghost shark (Callorhinchus milii, Holocephali, Chondrichthyes) as revealed by different immunohistochemical markers. AB - This article presents the first study on the glial architecture of a representative species of Holocephali, Callorhinchus milii (ghost shark). Holocephali are a small subclass of Chondrichthyes, with only a few extant genera, and those are considered to have a brain organization more similar to squalomorph sharks than to galeomorph sharks, skates, and rays. Three different astroglial markers--glial fibrillary acidic protein, S-100 protein, and glutamine synthetase (GS)--were investigated by immunohistochemical methods, applying both diaminobenzidine (DAB) and fluorescent techniques. They revealed similar glial structures, although most of them were detected by immunohistochemical reaction against GS and visualized by DAB. The predominant elements were radial ependymoglia spanning the area between the ventricular and meningeal surfaces, as in squalomorph sharks. Other similar features were the light appearance of myelinated neural tracts devoid of immunoreactivity, and the glial architecture of the reticular formation of the brain stem, cerebellum, and tectum, the latter with recognizable layers. The immunoreactivity of the vascular walls was similar; however, it is believed that different cell types form the blood-brain barrier in chimeras and in elasmobranchs. Some glial structures, however, resembled those of skates, rays, and galeomorph sharks. In C. milii astrocyte-like elements were observed in the telencephalon, using GS and S-100, although typical astrocyte rich regions were not found. In some areas, especially the telencephalon, not only endfeet but also cell bodies were observed to be attached to the meningeal surface, with processes extending into the brain substance. PMID- 18512703 TI - Patterns of angiogenic and hematopoietic gene expression during brown trout embryogenesis. AB - In this article, whole mount in situ hybridization is used to examine early blood vessel and blood cell development in the embryos of the brown trout Salmo trutta lacustris. cDNAs encoding for the angiogenic markers fli1 and flk1, and for the hematopoietic markers gata1 and gata2, were identified from an expressed sequence tag library of rainbow trout. Results show that fli1, flk1 and gata2 are activated in bilateral bands of the lateral trunk mesoderm before the onset of somitogenesis, shortly followed by gata1. These bands then converge toward the ventral midline to form the intermediate cell mass (ICM) (anterior ICM). Subsequent axial vasculogenesis and initial blood cell formation involve a clear spatial separation of fli1 and gata gene expression. Fli1 staining is most intense within the axial vessel (dorsal aorta, posterior cardinal vein) forming and lateral ICM cells, whereas binding of gata1 and gata2 probes becomes confined to the central portion of ICM cells beneath the dorsal aorta. This is followed by a first wave of angiogenesis, indicated by expression of fli1 and flk1. This gives rise to the intersegmental, dorsal longitudinal anastomotic and intestinal vessels. Further angiogenesis and hematopoiesis are activated in the "posterior ICM" of the tail. Here, the absence of gata1 indicates that hematopoiesis in this tissue generates myeloid rather than erythroid cells. The results supplement and validate previous, now historical morphological work in salmonids, thus aiding the elucidation of a comprehensive general scheme of angiogenic and hematopoietic development in the teleost embryo. PMID- 18512704 TI - Effects of incubation temperature on growth and performance of the veiled chameleon (Chamaeleo calyptratus). AB - I evaluated the effect of incubation temperature on phenotypes of the veiled chameleon, Chamaeleo calyptratus. I chose this species for study because its large clutch size (30-40 eggs or more) allows replication within clutches both within and among experimental treatments. The major research objectives were (1) to assess the effect of constant low, moderate, and high temperatures on embryonic development, (2) to determine whether the best incubation temperature for embryonic development also produced the "best" hatchlings, and (3) to determine how a change in incubation temperature during mid-development would affect phenotype. To meet these objectives, I established five experimental temperature regimes and determined egg survival and incubation length and measured body size and shape, selected body temperatures, and locomotory performance of lizards at regular intervals from hatching to 90 d, or just before sexual maturity. Incubation temperature affected the length of incubation, egg survival, and body mass, but did not affect sprint speed or selected body temperature although selected body temperature affected growth in mass independently of treatment and clutch. Incubation at moderate temperatures provided the best conditions for both embryonic and post-hatching development. The highest incubation temperatures were disruptive to development; eggs had high mortality, developmental rate was low, and hatchlings grew slowly. Changes in temperature during incubation increased the among-clutch variance in incubation length relative to that of constant temperature treatments. PMID- 18512705 TI - Temperament, ADHD and peer relations among schoolchildren: the mediating role of school bullying. AB - The goal of the study is to investigate the link between temperament, attention deficit hyperactivity disorder (ADHD) symptoms, social preference, bullying and victimization. The study sample consisted of 195 children attending the fourth and fifth grade (age 8-10), their mothers and their teachers. A multiinformant approach was used: mothers were interviewed about the temperament of their children; teachers were asked about ADHD symptoms; children responded to a peer nominations inventory designed to investigate their roles as bullies and/or victims and their social preferences (liked and disliked peers). A Structural Equation Model multigroup analysis (males and females) was used to test the relationships among variables hypothesized by the authors. The analysis showed that temperamental variables have a direct relation to ADHD symptoms, that ADHD has a direct relation to bullying behavior in males and to victimization in females and that bullies and victims are less well accepted than peers who belong to neither category. ADHD has only an indirect relation to social preference through the mediating role of school bullying. PMID- 18512706 TI - Metamorphic changes in localization of sugars in skin of the leopard frog, Rana pipiens. AB - A lectin histochemical study was carried out to determine the distribution of specific sugars in glycoconjugates within an important osmoregulatory organ, amphibian skin. Paraffin sections were made of Rana pipiens skin from dorsal and ventral regions of aquatic larvae in representative developmental stages as well as from several body regions of semiaquatic adult frogs. Sections were incubated with horseradish peroxidase (HRP)-conjugated lectins, which bind to specific terminal sugar residues of glycoconjugates. Such sites were visualized by DAB H2O2. The following HRP-lectins were used: UEA-1 for alpha-L-fucose, SBA for N acetyl-D-galactosamine, WGA for N-acetyl-beta-D-glucosamine, PNA for beta galactose, and Con A for alpha-mannose. We found that lectin binding patterns in larvae change during metamorphic climax as the skin undergoes extensive histological remodeling; this results in adult skin with staining patterns that are specific for each lectin and are similar in all body regions. Such findings in R. pipiens provide additional insight into the localization of molecules involved in osmoregulation in amphibian skin. PMID- 18512707 TI - No seasonal variation in the onset of symptoms of 445 patients with Wegener's granulomatosis. PMID- 18512708 TI - American College of Rheumatology 2008 recommendations for the use of nonbiologic and biologic disease-modifying antirheumatic drugs in rheumatoid arthritis. PMID- 18512709 TI - What we talk about when we talk about contraception. PMID- 18512710 TI - Improvement in patient-reported outcomes in a rituximab trial in patients with severe rheumatoid arthritis refractory to anti-tumor necrosis factor therapy. AB - OBJECTIVE: To assess the effects of treatment with rituximab plus methotrexate on patient-reported outcomes in patients with active rheumatoid arthritis (RA) who experienced inadequate response to anti-tumor necrosis factor therapy. METHODS: Patients with active RA were randomly assigned to rituximab (1,000 mg on days 1 and 15) or placebo. The primary end point was the proportion of patients with an American College of Rheumatology 20% response at week 24. Additional goals were to assess treatment effects on pain, fatigue, functional disability, health related quality of life, and disease activity by comparing mean changes between groups. The analysis was conducted in the intent-to-treat population. The proportion of patients who achieved the minimum clinically important difference on the Health Assessment Questionnaire (HAQ) disability index (DI), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), and Short Form 36 (SF 36) was determined. RESULTS: Rituximab patients had statistically significantly greater pain relief. The FACIT-F showed significantly greater improvement in rituximab patients than placebo patients from weeks 12 through 24. Mean improvement from baseline in functional disability (measured by the HAQ DI) was significantly greater in rituximab patients from weeks 8 to 24. The mean +/- SD change from baseline for the SF-36 Physical Component Score was 6.64 +/- 8.74 for rituximab patients and 1.48 +/- 7.32 for placebo patients (P < 0.0001). The mean change from baseline for the SF-36 Mental Component Score was 5.32 +/- 12.41 for rituximab patients and 2.25 +/- 12.23 for placebo patients (P = 0.0269). CONCLUSION: Rituximab produced rapid, clinically meaningful, and statistically significant improvements in patient-reported pain, fatigue, functional disability, health-related quality of life, and disease activity. These effects were sustained throughout the study. PMID- 18512712 TI - The 2008 American College of Rheumatology recommendations for the use of nonbiologic and biologic disease-modifying antirheumatic drugs in rheumatoid arthritis: where the rubber meets the road. PMID- 18512711 TI - Abnormal body composition phenotypes in older rheumatoid arthritis patients: association with disease characteristics and pharmacotherapies. AB - OBJECTIVE: To compare measures of body fat and lean mass and the prevalence of abnormal body composition phenotypes (sarcopenia, overfat, and sarcopenic obesity) in men and women with rheumatoid arthritis (RA) versus matched controls, and to explore the disease-related predictors of abnormal body composition in patients with RA. METHODS: A total of 189 men and women with RA and 189 age-, sex , and race-matched non-RA controls underwent dual-energy x-ray absorptiometry for measurement of total and regional body fat and lean mass. Continuous and categorical measures of body composition were compared between RA and control subjects by sex and according to categories of body mass index (BMI). Within the group of RA patients, demographic, lifestyle, and RA disease and treatment characteristics were compared for RA patients with healthy body composition versus those with abnormal body composition phenotypes. RESULTS: Compared with non-RA controls, RA status was significantly associated with greater odds of sarcopenia, overfat, and sarcopenic obesity in women, but not in men. Relative differences in body composition phenotypes between RA and control subjects were greatest for patients in the normal weight BMI category (<25 kg/m(2)). Among RA characteristics, increasing joint deformity, self-reported disability scores, C reactive protein levels, rheumatoid factor seropositivity, and a lack of current treatment with disease-modifying antirheumatic drugs were significantly associated with abnormal body composition. CONCLUSION: Abnormal body composition phenotypes are overrepresented in patients with RA, particularly in those in the normal weight BMI range. RA-associated disease and treatment characteristics contribute to this increase in abnormal body composition. PMID- 18512713 TI - Incidence of melanoma and other malignancies among rheumatoid arthritis patients treated with methotrexate. AB - OBJECTIVE: To determine cancer risk in a cohort of 459 rheumatoid arthritis (RA) patients treated with methotrexate in community practice. METHODS: All RA patients who started methotrexate prior to June 1986 and were attending 1 of 6 rheumatologists were studied. Demographic data were matched to the State Cancer Registry to identify all malignancies (except nonmelanoma skin cancer) for 1983 1998, and to the National Death Index to identify all deaths to the end of 1999. Followup started on the date when methotrexate was started and ended either on the last confirmed date on which the patient was seen by the rheumatologist or at death. Standardized incidence ratios (SIRs) were calculated using state population cancer rates stratified by sex, age (in 5-year groups), and calendar year. RESULTS: There were 4,145 person-years of followup (average 9.3 years). Eighty-seven malignancies were identified (14 before, 64 during, and 9 after the followup period). There was an estimated 50% excess risk of malignancy among methotrexate-exposed RA patients relative to the general population (SIR 1.5, 95% confidence interval [95% CI] 1.2-1.9), with a 3-fold increase in melanoma (SIR 3.0, 95% CI 1.2-6.2), a 5-fold increase in non-Hodgkin's lymphoma (SIR 5.1, 95% CI 2.2-10.0), and an almost 3-fold increase in lung cancer (SIR 2.9, 95% CI 1.6 4.8). CONCLUSION: Compared with the general population, methotrexate-treated RA patients have an increased incidence of melanoma, non-Hodgkin's lymphoma, and lung cancer. There may be a role for regular skin cancer screening for all RA patients, particularly those receiving immunosuppressive therapy. PMID- 18512714 TI - Effectiveness of the combination of a whole-blood interferon-gamma assay and the tuberculin skin test in detecting latent tuberculosis infection in rheumatoid arthritis patients receiving adalimumab therapy. AB - OBJECTIVE: To investigate QuantiFERON-tuberculosis Gold (QFT-G) assay and tuberculin skin test (TST) for latent tuberculosis (TB) infection (LTBI) in patients with rheumatoid arthritis (RA) treated with adalimumab. METHODS: We prospectively followed up 43 RA patients who received adalimumab therapy and underwent serial TSTs and QFT-G assays. TST was performed using Mantoux method and QFT-G assay was examined by measuring interferon-gamma levels in whole blood samples that were incubated with early secretary antigenic target-6 and culture filtrate protein 10. RESULTS: Before starting adalimumab therapy, 8 RA patients (18.6%) had positive and 35 (81.4%) had negative TST results. All 8 RA patients with positive TST results were diagnosed as LTBI and received isoniazid prophylaxis (INHP) 1 month before starting adalimumab therapy. None of these 8 RA patients developed active TB 2 years after completing INHP. A high rate (10 [37.0%] patients) of TST conversion was observed among 27 patients who had completed 12-month adalimumab therapy. Of these 10 patients with TST conversion, 2 patients had positive QFT-G results and 1 developed active TB disease. Among 17 RA patients who did not have TST conversion after 12-month adalimumab therapy, 1 patient who had a positive QFT-G result developed active TB disease. Of all 43 RA patients who received adalimumab therapy, 4 (9.3%) developed active TB after starting adalimumab therapy. CONCLUSION: The application of TST for detecting LTBI is limited in RA patients by the frequent presence of anergy. Combined QFT-G assay and TST can aid in detecting LTBI in RA patients receiving adalimumab therapy. PMID- 18512715 TI - Prevalence and associations of hallux valgus in a primary care population. AB - OBJECTIVE: To determine the population prevalence and examine factors associated with hallux valgus in a primary care population. METHODS: A questionnaire was mailed to all adults age >30 years registered with 2 general practices. Validated instruments assessed self-reported hallux valgus, nodal osteoarthritis, and knee pain. The questionnaire also asked about big toe pain, joint replacement, and history of osteoarthritis and rheumatoid arthritis. Hallux valgus prevalence was calculated and standardized by the source population in terms of age, sex, knee pain, osteoarthritis, and rheumatoid arthritis. A nested case-control study was undertaken and age-sex adjusted odds ratios (ORs) were calculated between hallux valgus and age, sex, body mass index, nodal osteoarthritis, knee pain, big toe pain, joint replacement, self-reported osteoarthritis, and self-reported rheumatoid arthritis, using a binary logistic regression model. RESULTS: A total of 13,684 questionnaires were mailed and 4,249 (32%) responses were received. The standardized prevalence of hallux valgus was 28.4%. Hallux valgus was associated with age (adjusted OR 1.61 per decade; 95% confidence interval [95% CI] 1.52 1.69), female sex (adjusted OR 2.64; 95% CI 2.26-3.08), nodal osteoarthritis (adjusted OR 1.66; 95% CI 1.26-2.17), knee pain (adjusted OR 1.96; 95% CI 1.65 2.32), big toe pain (adjusted OR 3.28; 95% CI 2.48-4.33), self-reported osteoarthritis (adjusted OR 1.41; 95% CI 1.15-1.72), and self-reported rheumatoid arthritis (adjusted OR 2.04; 95% CI 1.43-2.91). CONCLUSION: Hallux valgus is prevalent in the community and is associated with age, female sex, and components of generalized osteoarthritis such as nodal osteoarthritis, knee pain, big toe pain, and self-reported osteoarthritis. PMID- 18512716 TI - The effect of high strength static magnetic fields and ionizing radiation on gene expression and DNA damage in Caenorhabditis elegans. AB - Magnetic resonance imaging with high static magnetic fields (SMFs) has become widely used for medical imaging purposes because SMFs cause fewer genotoxic side effects than ionizing radiation (IR). However, the effect of exposure to high SMFs on global transcription is little understood. We demonstrate that genes involved in motor activity, actin binding, cell adhesion, and cuticles are transiently and specifically induced following exposure to 3 or 5 T SMF in the experimental model metazoan Caenorhabditis elegans. In addition, transient induction of hsp12 family genes was observed after SMF exposure. The small-heat shock protein gene hsp16 was also induced but to a much lesser extent, and the LacZ-stained population of hsp-16.1::lacZ transgenic worms did not significantly increase after exposure to SMFs with or without a second stressor, mild heat shock. Several genes encoding apoptotic cell-death activators and secreted surface proteins were upregulated after IR, but were not induced by SMFs. Real time quantitative RT-PCR analyses for 12 of these genes confirmed these expression differences between worms exposed to SMFs and IR. In contrast to IR, exposure to high SMFs did not induce DNA double-strand breaks or germline cell apoptosis during meiosis. These results suggest that the response of C. elegans to high SMFs is unique and capable of adjustment during long exposure, and that this treatment may be less hazardous than other therapeutic tools. PMID- 18512717 TI - Risk of unintended pregnancy among women with systemic lupus erythematosus. AB - OBJECTIVE: It is recommended that women with systemic lupus erythematosus carefully time their pregnancies, but little is known about use of contraception and risk of unintended pregnancy in this population. The goal of this study was to estimate the proportion of women with lupus at risk for unintended pregnancy. METHODS: We surveyed 309 women with lupus, ages 18-50 years, seen at the University of Pittsburgh lupus center between January and May 2007. RESULTS: Of the women surveyed, 212 (69%) completed the survey. In the past 3 months, 97 (46% of 212) had faced some risk of unintended pregnancy. Of these, 53 (55% of 97) reported >or=1 occasion on which they had unprotected sex and 22 (23% of 97) reported that in the last 3 months they had unprotected sex "most of the time." No women reported having used emergency contraception after unprotected sex. A desire to "discuss birth control with a health care provider at the lupus center" was reported by 22 (10% of 212) respondents and 16 (17% of 94) women 400,000 rheumatologically oriented manuscripts (0.026%). Very few manuscripts consisted of original research studies. Nonmaleficence (66%) was the most common theme, whereas justice represented the least frequently addressed ethical issue (12%). The differences in the proportions of each ethical principle reached statistical significance (Q = 73.8, P < 0.0001). Only 8 articles addressed >2 ethical principles. Discussion touching on autonomy and nonmaleficence frequently appeared in the same article (Pearson's chi(2) = 14.9, P < 0.001). CONCLUSION: Despite the frequency of ethical issues while caring for patients, few reports within the rheumatic disease literature have focused on ethical issues. Further work should ascertain the degree to which the literature addresses the ethical questions in rheumatology. PMID- 18512719 TI - Development and validation of a needs-based quality of life instrument for osteoarthritis. AB - OBJECTIVE: To develop and validate a disease-specific quality of life (QOL) measure for osteoarthritis (OA), the OAQoL, using the needs-based conceptual model. METHODS: In the first phase of this study, in-depth, semistructured interviews were conducted with 44 OA patients to explore the issues associated with impact of OA and to derive items for a draft OAQoL questionnaire. In phase 2, 17 OA patients were interviewed on the relevance, clarity, and ease of completion of the measure in structured interviews. In phase 3, the draft questionnaire was mailed to 635 patients to test the psychometric properties of the questionnaire using Rasch analysis. Test-retest assessment of the revised questionnaire was performed in phase 4 by mailing the questionnaire to an additional 201 participants, with a second questionnaire repeated 2 weeks later. RESULTS: A 38-item draft measure was devised during phase 1 and mailed in phase 2. Rasch analysis of the draft questionnaire (n = 259) indicated initial misfit, which was rectified with the removal of 13 problematic items (chi(2)[75] = 83.602, P = 0.232). For the test-retest assessment (n = 60), 3 additional items were removed, leaving a 22-item OAQoL that demonstrated good fit to the Rasch model (chi(2)[44] = 44.559, P = 0.533) with excellent test-retest correlation (rho = 0.925, P < 0.001; z = -0.06, P = 0.995). CONCLUSION: The OAQoL is a simple and easy to use 22-item unidimensional questionnaire developed specifically to assess the impact of OA on QOL. The measure has been developed as a true patient based questionnaire and demonstrates good psychometric properties, including test retest reliability. PMID- 18512720 TI - The impact of momentary pain and fatigue on physical activity in women with osteoarthritis. AB - OBJECTIVE: To examine the daily life patterns of both pain and fatigue symptoms and objective physical activity (using ambulatory monitoring) in women with symptomatic lower extremity osteoarthritis (OA), and to evaluate how momentary symptoms impact physical activity levels. METHODS: Sixty women age >or=55 years (40 with knee or hip OA and 20 matched controls) participated in an observational study involving 2 laboratory visits and a 5-day home data collection period. During the home period physical activity levels were assessed continuously, and symptoms were inputted 6 times a day into an enhanced accelerometer at prespecified time points. RESULTS: In the OA group as compared with the control group over the 5-day period, average physical activity was significantly lower (P = 0.02) and peak physical activity tended to be lower (P = 0.06). Although pain and fatigue overall were of moderate severity in this cohort, fatigue escalated throughout each day. In a hierarchical linear model, fatigue was most strongly associated with physical activity (beta = -30.1, P < 0.0001). Pain was more weakly associated with physical activity and in the direction opposite to what was hypothesized (beta = 16.9, P = 0.04). CONCLUSION: Momentary reports of fatigue negatively predicted physical activity levels and were much more strongly related to physical activity than momentary pain. In order to help women with knee or hip OA manage symptoms and become more physically active, it may be important to emphasize fatigue management. PMID- 18512721 TI - Defining appropriate outcome measures in pulmonary arterial hypertension related to systemic sclerosis: a Delphi consensus study with cluster analysis. AB - OBJECTIVE: Outcome measures for pulmonary arterial hypertension associated with systemic sclerosis (PAH-SSc) are only partially validated. The aim of the present study was to establish an expert consensus regarding which outcome measures are most appropriate for clinical trials in PAH-SSc. METHODS: Sixty-nine PAH-SSc experts (rheumatologists, cardiologists, pulmonologists) rated a list of disease domains and measurement tools in an Internet-based 3-stage Delphi consensus study. In stages 2 and 3, the medians of domains and measurement tools and frequency distributions of ratings, along with requests for re-ratings, were distributed to respondents to provide feedback. A final score of items was identified by means of cluster analysis. RESULTS: The experts judged the following domains and tools as most appropriate for randomized controlled trials in PAH-SSc: lung vascular/pulmonary arterial pressure and cardiac function both measured by right heart catheterization and echocardiography, exercise testing measured by 6-minute walking test and oxygen saturation at exercise, severity of dyspnea measured on a visual analog scale, discontinuation of treatment measured by (serious) adverse events, quality of life/activities of daily living measured by the Short Form 36 and Health Assessment Questionnaire disability index, and global state assessed by physician measured by survival. CONCLUSION: Among experts in PAH-SSc, a core set of outcome measures has been defined for clinical trials by Delphi consensus methods. Although these outcome measures are recommended by this expert group to be used as an interim tool, it will be necessary to formally validate the present measures, as well as potential research measures, in further studies. PMID- 18512722 TI - Assessment of the item selection and weighting in the Birmingham vasculitis activity score for Wegener's granulomatosis. AB - OBJECTIVE: To assess the Birmingham Vasculitis Activity Score for Wegener's Granulomatosis (BVAS/WG) with respect to its selection and weighting of items. METHODS: This study used the BVAS/WG data from the Wegener's Granulomatosis Etanercept Trial. The scoring frequencies of the 34 predefined items and any "other" items added by clinicians were calculated. Using linear regression with generalized estimating equations in which the physician global assessment (PGA) of disease activity was the dependent variable, we computed weights for all predefined items. We also created variables for clinical manifestations frequently added as other items, and computed weights for these as well. We searched for the model that included the items and their generated weights yielding an activity score with the highest R(2) to predict the PGA. RESULTS: We analyzed 2,044 BVAS/WG assessments from 180 patients; 734 assessments were scored during active disease. The highest R(2) with the PGA was obtained by scoring WG activity based on the following items: the 25 predefined items rated on >or=5 visits, the 2 newly created fatigue and weight loss variables, the remaining minor other and major other items, and a variable that signified whether new or worse items were present at a specific visit. The weights assigned to the items ranged from 1 to 21. Compared with the original BVAS/WG, this modified score correlated significantly more strongly with the PGA. CONCLUSION: This study suggests possibilities to enhance the item selection and weighting of the BVAS/WG. These changes may increase this instrument's ability to capture the continuum of disease activity in WG. PMID- 18512723 TI - Occupational physical activities and long-term functional and radiographic outcomes in patients with ankylosing spondylitis. AB - OBJECTIVE: We sought to identify specific occupational activities associated with functional limitations and radiographic damage in patients with longstanding ankylosing spondylitis (AS). METHODS: We asked patients diagnosed with AS for >or=20 years to report all past occupations, which we mapped to specific physical activities using the Occupational Information Network, which is the US Department of Labor job classification database. For each occupation reported, we obtained ratings for 13 physical abilities of the worker and 13 aspects of the work environment or work tasks (work context) thought to be most relevant to patients with AS. Averages for each measure, weighted by the number of years in each job, were related to the degree of functional limitation as assessed by the Bath AS Functional Index (BASFI) and to the extent of spinal radiographic damage as assessed by the Bath AS Radiology Index for the spine (BASRI-s). RESULTS: Among 397 patients, those with a history of jobs requiring dynamic flexibility (the ability to repeatedly bend, stretch, twist, or reach) had more functional limitations than those whose past jobs required little or no dynamic flexibility (adjusted mean BASFI score 48.3 in the top quartile versus 38.1 in all others). Those whose past jobs required more dynamic flexibility, extent flexibility, and exposure to whole body vibration also had significantly higher BASRI-s scores. CONCLUSION: Bending, twisting, and stretching are the occupational activities associated with greater functional limitations and radiographic damage in patients with longstanding AS. Exposure to whole body vibration was also associated with more radiographic damage. PMID- 18512724 TI - The relationship between affect balance style and clinical outcomes in fibromyalgia. AB - OBJECTIVE: Affective balance, relative levels of negative affect (NA) and positive affect (PA), better describes emotional functioning than NA or PA alone. Affect balance styles and their relationship to clinical outcomes were compared between patients with fibromyalgia (FM) and controls. METHODS: FM patients (n = 79) were compared with patients with other medical conditions (controls; n = 92). Patients underwent a physical examination, completed questionnaires, and were screened for clinical disorders such as depression, with diagnoses confirmed by structured interview. Affect balance style categories were calculated as follows: healthy (high PA/low NA), low (low PA/low NA), reactive (high PA/high NA), and depressive (low PA/high NA). RESULTS: Compared with controls, FM patients had lower levels of PA (P = 0.0031; P values are adjusted for multiple testing), higher levels of NA (P = 0.0061), lower levels of functioning (P < 0.0001), and more clinical disorders (P = 0.0031). Groups differed regarding affect balance style (P = 0.0061), with FM patients being more likely than controls to be categorized as depressive (odds ratio 5.60) and reactive (odds ratio 3.81). FM patients and controls with reactive and depressive affect balance styles reported poorer functioning (P < 0.0001) compared with patients with healthy affect balance style. Finally, there was an association between affect balance style and psychiatric comorbidity (P < 0.0001), with patients with depressive and reactive affect balance styles having a 9.00 and 4.75 odds ratio, respectively, of having psychiatric comorbidity compared with patients with healthy affect balance style. CONCLUSION: Depressive (low PA, high NA) and reactive (high PA, high NA) affect balance styles were predominant in FM patients and related to poor functioning and psychiatric comorbidity. PMID- 18512725 TI - Findings of extensive variation in the types of outcome measures used in hip and knee replacement clinical trials: a systematic review. AB - OBJECTIVE: To describe the extent of variation in outcome measure usage in hip and knee replacement randomized trial literature, and to summarize this variation in the context of the International Classification of Functioning, Disability, and Health (ICF) conceptual model created by the World Health Organization (WHO). METHODS: We used a defined search strategy in Medline and EMBase databases to identify articles published from January 2000 to February 2007. Studies were reviewed if they were randomized trials with a >or=6-week followup and if they used noninvasive outcome measures of impaired joint function or whole-person limitations in daily activities or functional status. The WHO ICF model was used to categorize outcome measures. RESULTS: Of 972 studies, 160 were included for review. Of these, 82 were conducted on patients with hip replacements, 75 on patients with knee replacements, and 3 on patients with both. The most common outcome measure in knee trials was the American Knee Society score (used in 48% of reviewed studies), and in hip trials was the Harris hip score (52.4%). At least 20 different outcome measures were used in the hip trials, and at least 14 different measures were used in knee trials. The primary outcome was identified in only 24% of trials. CONCLUSION: We found extensive variation in outcome measures across trials and saw inconsistency across the components of the WHO ICF model. To improve interpretability, future work should determine whether consensus can be developed for a standardized set of outcome measures for hip and knee replacement trials. PMID- 18512726 TI - A 73-year-old woman with rheumatoid arthritis and shortness of breath. PMID- 18512727 TI - Mycobacterium kansasii tenosynovitis in a rheumatoid arthritis patient with long term therapeutic immunosuppression. PMID- 18512728 TI - Agents used for chemoprevention of prostate cancer may influence PSA secretion independently of cell growth in the LNCaP model of human prostate cancer progression. AB - BACKGROUND: The aim of this study was to evaluate the inhibitory growth effects of different potential chemopreventive agents in vitro and to determine their influence on PSA mRNA and protein expression with an established screening platform. METHODS: LNCaP and C4-2 cells were incubated with genistein, seleno-L methionine, lycopene, DL-alpha-tocopherol, and trans-beta-carotene at three different concentrations and cell growth was determined by the MTT assay. PSA mRNA expression was assessed by quantitative real-time RT-PCR and secreted PSA protein levels were quantified by the microparticle enzyme immunoassay. RESULTS: Genistein, seleno-l-methionine and lycopene inhibited LNCaP cell growth, and the proliferation of C4-2 cells was suppressed by seleno-L-methionine and lycopene. PSA mRNA expression was downregulated by genistein in LNCaP but not C4-2 cells. No other compound tested altered PSA mRNA expression. PSA protein expression was downregulated by genistein, seleno-L-methionine, DL-alpha-tocopherol in LNCaP cells. In C4-2 cells only genistein significantly reduced the secretion of PSA protein. CONCLUSIONS: In the LNCaP progression model PSA expression depends on the compound, its concentration and on the hormonal dependence of the cell line used and does not necessarily reflect cell growth or death. Before potential substances are evaluated in clinical trials using PSA as a surrogate end point marker, their effect on PSA mRNA and protein expression has to be considered to correctly assess treatment response by PSA. PMID- 18512729 TI - Osteoblasts-derived BMP-2 enhances the motility of prostate cancer cells via activation of integrins. AB - BACKGROUND: Bone metastases are common complications of prostate cancer cells. The bone morphogenetic protein-2 (BMP-2) is constitutively secreted by osteoblasts and plays a key role in bone formation. Integrins are the major adhesive molecules in mammalian cells, and has been associated with cancer cells metastasis to bone. The aim of this study was to investigate whether osteoblast derived BMP-2 is associated with prostate cancer metastasis. METHOD: Cancer cells migration activity was examined using the Transwell assay. The ERK and AKT phosphorylation was examined by using Western blot method. The siRNA was used to inhibit the expression of BMP-2. The cell surface expression of integrins was examined by using flow cytometry. A transient transfection protocol was used to examine NF-kappaB activity. RESULTS: We found that osteoblast conditioned medium (OBCM) increased the migration and cell surface expression of beta1 or beta 3 integrin in human prostate cancer cells. beta1 or beta 3 integrin monoclonal antibodies or siRNA against beta1 or beta 3 integrin inhibited the OBCM-induced increase the migration of prostate cancer cells. BMP-2 siRNA specifically reduced the OBCM-induced migration and integrins upregulation. BMP-2 siRNA also suppressed the OBCM-induced ERK, AKT and NF-kappaB activation. CONCLUSIONS: This study suggest that the osteoblast-derived BMP-2 act through Akt and ERK, which in turn activates IKK alpha/beta and NF-kappaB, resulting in the activations of beta1 and beta 3 integrins and contributing the migration of prostate cancer cells. PMID- 18512730 TI - Translationally regulated C/EBP beta isoform expression upregulates metastatic genes in hormone-independent prostate cancer cells. AB - BACKGROUND: C/EBP beta is a transcription factor regulating key biological processes including cellular growth and differentiation and its increased expression correlates with tumor invasiveness. Recently, the increased expression of C/EBP beta was reported in proliferative inflammatory atrophy of the prostate, associating with increased COX-2 expression and androgen receptor (AR) downregulation. METHODS: C/EBP beta expression was determined in DU-145, PC-3 and LNCaP cells by immunoblotting. Transient transfection of C/EBP beta expression vectors was performed to investigate translational regulation of its isoform expression. Reporter gene analysis was performed to investigate transcriptional activity of C/EBP beta on metastatic gene expression. RESULTS: We determined that transcriptionally active, full-length C/EBP beta isoforms were dominantly expressed in hormone-independent DU-145 and PC-3 cells, while transcription repressing truncated isoform was dominant in hormone-dependent LNCaP cells. Our results further showed lack of full-length isoform expression from the transiently transfected C/EBP beta expression vector in LNCaP cells compared to that in PC-3 cells transfected with the same vector, while the expression of truncated isoform was comparable in both cell lines. Interestingly, however, the most upstream initiation site A null mutation restored translation of full-length isoform in LNCaP cells. These results suggest that full-length C/EBP beta isoform expression in LNCaP cells may be suppressed at the upstream initiation sites, likely at site A. Most importantly, C/EBP beta overexpression significantly upregulated promoter activities of IL-8, COX-2, and anti-apoptotic Bfl-1 genes. CONCLUSIONS: Our study demonstrates that C/EBP beta is an important transcription factor upregulating metastatic gene expression and that its isoform expression is differentially regulated at the translational level in prostate cancer cells. PMID- 18512731 TI - Inhibition of prostate efferent neurotransmission by amikacin. AB - BACKGROUND: It has been suggested that manipulation of the autonomic nerve supply to the prostate leads to loss of functional and structural integrity of the gland, and that these changes may be useful in treating prostatic diseases. This study investigates the effect of amikacin on prostate efferent neurotransmission in vitro, in both rat and human prostate samples. METHODS: Prostate samples, obtained from male Wistar rats and 8 patients undergoing endoscopic surgery for benign prostatic hyperplasia, were studied by measurement of isometric contraction induced by electrical field stimulation (EFS), noradrenalin, carbachol, serotonin and ATP, in the presence or absence of amikacin 10(-3) M in a low-Ca medium. RESULTS: Amikacin significantly reduced EFS-induced contraction of isolated rat and human prostate samples by 45 +/- 6.5% (P < 0.01) and 47 +/- 6% (P < 0.01), respectively. Contraction was restored after addition of calcium chloride 2 x 10(-3) M. Amikacin-induced relaxation in rat prostate samples was greater than the effect of magnesium and weaker than those of prazosin and verapamil, but comparable in extent with the latter. No significant effect was observed on rat prostate contractile response to noradrenaline as to ATP in the presence of amikacin. Rat prostate contraction to carbachol and serotonin was inhibited by 35 +/- 11% (P < 0.05) and 59 +/- 11.7% (P < 0.01), respectively, after addition of amikacin 10(-3) M. CONCLUSIONS: Amikacin reduces in vitro both rat and human prostate contraction elicited by pre-junctional stimulation, but does not affect prostate contraction evoked by post-junctional stimulation. Our results indicate that amikacin affects prostatic efferent neurotransmission. PMID- 18512732 TI - Effects of Eg5 knockdown on human prostate cancer xenograft growth and chemosensitivity. AB - OBJECTIVES: Microtubular inhibitors, including docetaxel, are active cytotoxics in many cancers, including prostate cancer (CaP). The Eg5 gene, a member of the kinesin-5 family, plays critical roles in proper mitotic spindle function, and is a potential microtubule-related target for proliferating cancer cells. To investigate the functional activities of Eg5 in CaP, we used an antisense oligonucleotide (ASO) targeting Eg5 to assess the potency and anti-cancer activity of Eg5 ASO treatment for androgen-independent CaP cells in vitro and in vivo. RESULTS: PC3 cells express higher Eg5 protein and mRNA levels compared to LNCaP cells. In both cell lines, Eg5 ASO treatment reduced mRNA and protein levels in a dose-dependent manner and a complete reduction of Eg5 protein levels was observed at 100 nM. Dose-dependent inhibition in cell growth, potent G2/M phase arrest, and increases in apoptotic sub-G1 fraction were also observed using Eg5 ASO. Surprisingly, low dose Eg5 ASO significantly antagonized cytotoxic effects of paclitaxel. In vivo, Eg5 ASO monotherapy significantly reduced both LNCaP and PC-3 tumor growth but combination treatment with paclitaxel did not yield additive benefits. CONCLUSIONS: These findings suggest that while Eg5 is a potential target to delay androgen-independent CaP growth, combination treatment with paclitaxel may not be desirable. PMID- 18512733 TI - No association between tagging SNPs of SNARE complex genes (STX1A, VAMP2 and SNAP25) and schizophrenia in a Japanese population. AB - Abnormalities in neural connections and the neurotransmitter system appear to be involved in the pathophysiology of schizophrenia. The soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) complex, which consists of Syntaxin1A, vesicle-associated membrane protein 2 (VAMP2) and synaptosomal associated protein 25 kDa (SNAP25), plays an important role in the neurotransmitter system, and is therefore an attractive place to search for candidate genes for schizophrenia. We conducted a two-stage genetic association analysis of Syntaxin1A (STX1A), VAMP2 and SNAP25 genes with schizophrenia (first set screening samples: 377 cases and 377 controls, second-set confirmation samples: 657 cases and 527 controls). Based on the linkage disequilibrium, 40 SNPs (STX1A, 8 SNPs; VAMP2, 3 SNPs; SNAP25, 29 SNPs) were selected as 'tagging SNPs'. Only nominally significant associations of an SNP (rs12626080) and haplotype (rs363014 and rs12626080) in SNAP25 were detected in the first-set screening scan. To validate this significance, we carried out a replication analysis of these SNP and haplotype associations in second-set samples with a denser set of markers (including five additional SNPs). However, these associations could not be confirmed in the second-set analysis. These results suggest that the SNARE complex-related genes do not play a major role in susceptibility to schizophrenia in the Japanese population. PMID- 18512734 TI - Remission of CVB3-induced viral myocarditis by in vivo Th2 polarization via hydrodynamics-based interleukin-4 gene transfer. AB - BACKGROUND: Regulation of Th polarization was critical for the prevention of Coxsackievirus B3 (CVB3) induced myocarditis. In the present study, interleukin (IL)-4 was over-expressed by hydrodynamics-based gene transfection (HGT) to induce the in vivo Th2 bias and evaluate the influence of Th polarization on the pathogenesis of CVB3-myocaditis. METHODS: IL-4 expressing plasmid was delivered into BALB/c mice by HGT after CVB3 infection. In vivo Th polarization was evaluated by detecting expression of Th1/Th2 cytokine, antibody isotype and Th related transcription factor, as well as the proliferation of CD8(+) T cells. The severity of myocarditis was assessed by weight loss, serological index of myocarditis, pathological feature, as well as survival rate. RESULTS: HGT of IL-4 plasmid resulted in high-level and long-lasting expression of IL-4 in different organs, which rescued mice from severe heart inflammation and death. This may due to the induction of a Th2-bised immune response specified with decreased expression of tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma but increased expression of IL-10 and IL-4 in serum and heart tissue, more IL-4 but less IFN-gamma secreting splenic CD4+ T cells, an immunoglobulin G1 isotype switch, increased expression of GATA-3 and low proliferation of CD8+ T cells, without significant change of virus titer in heart tissue. CONCLUSIONS: CVB3 induced myocarditis could be remitted through in vivo Th2 polarization, which has implications for our understanding of the role of Th2 population in immunity to CVB3 infection and for the development of new therapies for CVB3-myocarditis. PMID- 18512735 TI - Gender differences in metabolic monitoring of second-generation antipsychotic prescription. AB - OBJECTIVE: To explore whether guidelines for metabolic monitoring of patients receiving second-generation antipsychotics (SGAP) are respected in clinical practice, and whether patients' and prescribers' characteristics are associated with adequacy of metabolic monitoring. METHOD: Survey questionnaires were mailed to all state hospital psychiatrists practising in South-Western France. Psychiatrists were asked to give information on the baseline metabolic screening of the last patient seen with first prescription of a SGAP. Adequacy of metabolic monitoring was assessed according to published guidelines. RESULTS: No metabolic parameter was assessed before SGAP onset in more than one out of four patients, and fewer than 5% had a complete baseline assessment of metabolic parameters. The number of parameters assessed was markedly lower in women compared to men. CONCLUSION: Efforts should focus on initial training as well as on continuing medical education in order to improve level of awareness about the need for adequate baseline screening monitoring, and about the fact that women treated with SGAP are also at risk of metabolic disturbances. PMID- 18512737 TI - Intimate coupling of photocatalysis and biodegradation in a photocatalytic circulating-bed biofilm reactor. AB - Coupling advanced oxidative pretreatment with subsequent biodegradation demonstrates potential for treating wastewaters containing biorecalcitrant and inhibitory organic constituents. However, advanced oxidation is indiscriminate, producing a range of products that can be too oxidized, unavailable for biodegradation, or toxic themselves. This problem could be overcome if advanced oxidation and biodegradation occurred together, an orientation called intimate coupling; then, biodegradable organics are removed as they are formed, focusing the chemical oxidant on the non-biodegradable fraction. Intimate coupling has seemed impossible because the conditions of advanced oxidation, for example, hydroxyl radicals and sometimes UV-light, are severely toxic to microorganisms. Here, we demonstrate that a novel photocatalytic circulating-bed biofilm reactor (PCBBR), which utilizes macro-porous carriers to protect biofilm from toxic reactants and UV light, achieves intimate coupling. We demonstrate the viability of the PCBBR system first with UV only and acetate, where the carriers grew biofilm and sustained acetate biodegradation despite continuous UV irradiation. Images obtained by scanning electron microscopy and confocal laser scanning microscopy show bacteria living behind the exposed surface of the cubes. Second, we used slurry-form Degussa P25 TiO2 to initiate photocatalysis of inhibitory 2,4,5-trichlorophenol (TCP) and acetate. With no bacterial carriers, photocatalysis and physical processes removed TCP and COD to 32% and 26% of their influent levels, but addition of biofilm carriers decreased residuals to 2% and 4%, respectively. Biodegradation alone could not remove TCP. Photomicrographs clearly show that biomass originally on the exterior of the carriers was oxidized (charred), but biofilm a short distance within the carriers was protected. Finally, we coated TiO2 directly onto the carrier surface, producing a hybrid photocatalytic-biological carrier. These carriers likewise demonstrated the concept of photocatalytic degradation of TCP coupled with biodegradation of acetate, but continued TCP degradation required augmentation with slurry-form TiO2. PMID- 18512738 TI - Low-voltage transmission electron microscopy reveals SV40 viral particles within secretory granules in pancreatic cells. AB - Novel approach in low voltage transmission electron microscopy (TEM) has revealed the presence of SV40 viral like particles in the secretory zymogen granules and in spherical membrane-bound dense bodies of SV40 infected pancreatic cells. The presence of SV40 antigen in these cellular compartments was confirmed by immunocytochemistry of the VP1 antigen. Visualization of the viral particles was only possible by examining ultrathin tissue sections with low-voltage TEM that significantly enhances imaging contrast. Results indicate that following infection of the cell entry and trafficking of the viral particles are present in unique cellular compartments such as ER, dense bodies, and secretory granules. PMID- 18512739 TI - SEM analysis of enamel surface treated by Er:YAG laser: influence of irradiation distance. AB - BACKGROUND: Depending on the distance of laser tip to dental surface a specific morphological pattern should be expected. However, there have been limited reports that correlate the Er:YAG irradiation distance with dental morphology. PURPOSE: To assess the influence of Er:YAG laser irradiation distance on enamel morphology, by means of scanning electron microscopy (SEM). METHODS: Sixty human third molars were employed to obtain discs (approximately =1 mm thick) that were randomly assigned to six groups (n=10). Five groups received Er:YAG laser irradiation (80 mJ/2 Hz) for 20 s, according to the irradiation distance: 11, 12, 14, 16, or 17 mm and the control group was treated with 37% phosphoric acid for 15 s. The laser-irradiated discs were bisected. One hemi-disc was separated for superficial analysis without subsequent acid etching, and the other one, received the phosphoric acid for 15 s. Samples were prepared for SEM. RESULTS: Laser irradiation at 11 and 12 mm provided an evident ablation of enamel, with evident fissures and some fused areas. At 14, 16 and 17 mm the superficial topography was flatter than in the other distances. The subsequent acid etching on the lased surface partially removed the disorganized tissue. CONCLUSIONS: Er:YAG laser in defocused mode promoted slight morphological alterations and seems more suitable for enamel conditioning than focused irradiation. The application of phosphoric acid on lased-enamel surface, regardless of the irradiation distance, decreased the superficial irregularities. PMID- 18512740 TI - 3D reconstruction of high-resolution STED microscope images. AB - Tackling biological problems often involves the imaging and localization of cellular structures on the nanometer scale. Although optical super-resolution below 100 nm can be readily attained with stimulated emission depletion (STED) and photoswitching microscopy methods, attaining an axial resolution <100 nm with focused light generally required the use of two lenses in a 4Pi configuration or exceptionally bright photochromic fluorophores. Here, we describe a simple technical solution for 3D nanoscopy of fixed samples: biological specimens are fluorescently labeled, embedded in a polymer resin, cut into thin sections, and then imaged via STED microscopy with nanoscale resolution. This approach allows a 3D image reconstruction with a resolution <80 nm in all directions using available state-of-the art STED microscopes. PMID- 18512741 TI - Shannon's entropy and fractal dimension provide an objective account of bone tissue organization during calvarial bone regeneration. AB - The regeneration of compact bone involves the deposition of a poorly organized connective tissue template that remodels into compact lamellar bone. An objective description of this process is difficult because classical histomorphometry is unable to correctly characterize qualitative changes in tissue complexity. In this study, we demonstrated the use of two distinct methods of image texture analysis, the Shannon's entropy [standard error (SE)], and the fractal dimension (FD) to characterize the formation and remodeling of newly formed compact bone within two different polyanionic collagen-elastin matrices. The matrices were implanted in defects created into parietal bones of rats. The SE and FD were calculated for histological images of the experimental groups collected 3, 7, 15, 30, 60, and 365 days postsurgery and for the original bone only at day 365. Results showed that the SE and the FD initially increased and then diminished for all groups from day 3 to day 365 approaching the values of the original bone. These results are consistent with poor tissue organization during early osteogenesis that remodels into an organized lamellar structure, showing that these methods can be valuable tools to describe bone tissue remodeling during the regeneration process of compact bones. PMID- 18512742 TI - Characterization of shapes for use in classification of starch grains images. AB - As tradition Chinese herbal medicine becomes increasingly popular, there is an urgent need for efficient and accurate methods for the authentication of the Chinese Materia Medica (CMM) used in the herbal medicine. In this work, we present a denoising filter and introduce the use of chord length distribution (CLD) for the classification of starch grains in microscopic images of Chinese Materia Medica. Our simple denoising filter is adaptive to the background and is shown to be effective to remove noise, which appears in CMM microscopic starch grains images. The CLD is extracted by considering the frequency of the chord length in the binarized starch grains image, and we shall show that the CLD is an efficient and effective characterization of the starch grains. Experimental results on 240 starch grains images of 24 classes show that our method outperforms benchmark result using the current state-of-the-art method based on circular size distribution extracted by morphological operators at much higher computational cost. PMID- 18512743 TI - Ab-initio multireference study of an organic mixed-valence Spiro molecular system. AB - The electronic structure and some electron transfer properties of a model mixed valence Spiro molecular cation have been investigated at CAS-SCF, CAS+S, and CAS+SD levels starting from canonical and localized orbitals, using SZ, DZ, and TZP basis sets. The potential energy surfaces of the adiabatic ground and the lowest three excited electronic states have been computed, within a two-state model, and a double-well potential has been obtained for the ground electronic state. We have demonstrated the low coupling interaction between the two redox moieties of this molecular cation by following the charge localization/delocalization in the valence pi system through the reaction coordinate of the intramolecular charge transfer. The effect of dynamical correlation, using either localized or canonical orbitals, was found to be crucial for a quantitative description of the electronic structure and some important electron transfer parameters of this mixed-valence system. PMID- 18512744 TI - Diffusion-weighted imaging and magnetization transfer imaging of tardive and edentulous orodyskinesia. AB - Oral dyskinesias occur in elderly individuals in relation to drug use (tardive dyskinesia, TD) or edentulousness (edentulous orodyskinesia, EOD) but their characterization remains incomplete. Our aim was to investigate whether magnetic resonance techniques such as diffusion-weighted imaging (DWI) and magnetization transfer imaging (MTI) of the brain could be used to differentiate dyskinetic patients from control subjects. Eight drug-treated patients with TD, 12 EOD patients, 8 drug-treated patients without TD, and 10 control subjects were recruited and examined by DWI and MTI. Measurements in the caudate nucleus, putamen, and globus pallidus yielded globally different apparent diffusion coefficient (ADC) values between drug treated patients with TD and control subjects but the magnetization transfer ratios showed no significant variations. The discrimination between dyskinetic patients and control subjects offered by ADC values was however slightly poorer than the discrimination offered by the previously published choline/creatine ratios measured by MR spectroscopy in the basal ganglia. The results are consistent with the pathophysiological hypothesis of damage to cholinergic interneurons. PMID- 18512745 TI - Unverricht-Lundborg disease-a misnomer? PMID- 18512746 TI - Philadelphia Geriatric Morale Scale in essential tremor: a population-based study in three Spanish communities. AB - Essential tremor (ET) is associated with both functional disability and depression. Each could contribute to a poor sense of well-being and low morale. We hypothesized that morale would be lower in ET cases than controls. Using a population-based, cross-sectional design, morale was assessed in 187 ET cases and 561 matched controls living in three communities in central Spain using the Philadelphia Geriatric Center Morale Scale (PGCMS) (range = 0 [low morale]-17), which included three-dimensions of psychological well-being: agitation, lonely dissatisfaction, and attitude toward own aging. The PGCMS score was lower in ET cases than controls (9.41 +/- 3.21 vs. 10.39 +/- 2.92, P < 0.001), as were the Agitation subscore (3.17 +/- 1.71 vs. 3.78 +/- 1.67, P< 0.001) and Lonely Dissatisfaction subscore (3.75 +/- 1.34 vs. 4.02 +/- 1.24, P < 0.05). Nearly one half of the ET cases were classified as having low morale compared with only one third of controls (P = 0.006). In a linear regression analysis adjusting for demographic factors and multiple comorbid conditions, ET cases had a lower log PGCMS score than controls (P < 0.001). Exclusion of participants on antidepressant medication did not change the results. Our results indicate that morale is significantly lower in community-dwelling ET cases than in matched controls. This lower morale could in part be a proxy for mild, untreated depression. It therefore seems important to detect and then possibly treat this problem to improve the psychological well-being of patients with this disease. PMID- 18512747 TI - Effect of external cueing on gait in Huntington's disease. AB - In Huntington's disease (HD) patients, gait is characterized by a timing disorder with marked intraindividual variability in temporal gait parameters (caused by the presence of both hyperkinetic and hypokinetic features). We sought to determine the influence of use of a metronome on gait parameters in patients simultaneously performing motor or cognitive tasks that required attentional resources. The objective is to evaluate the influence of rhythmic cues on gait interference during self-regulated walking and a dual task paradigm in HD. Fifteen HD patients and 15 paired controls were asked to walk and simultaneously perform another motor task (carrying a tray with four full glasses) or a cognitive task (counting backwards). We evaluated the effect of a metronome (set at 100% and 120% of the subject's self-determined cadence) in three different task conditions (gait alone, gait + motor task, gait + cognitive task). The use of auditory cues during free gait and dual tasks did not improve kinematic parameters in HD patients, in contrast to the situation for control subjects (improvement in gait speed and cadence but not stride length when the metronome was set at 120% in all conditions). HD patients have difficulty in synchronizing their footsteps with a metronome, mainly due to attentional deficits. PMID- 18512748 TI - Fatigue and daytime somnolence in Machado Joseph Disease (spinocerebellar ataxia type 3). PMID- 18512749 TI - REM behavior disorder, hallucinations and cognitive impairment in Parkinson's disease: a two-year follow up. AB - In Parkinson's disease (PD) the presence of REM parasonnias as REM Behaviour Disorder (RBD) or vivid dreams/nightmares, is recognized as largely associated with hallucinations, even if the risk of the development of hallucinations seem not to depend on how long the REM parasomnias had been occurring. The aim of this study was to establish if RBDs occurring earlier than hallucinations in PD are predictive of cognitive impairment development. Three groups of PD patients: i) group 1, without RBD and without hallucinations; ii) group 2, with RBD but without hallucinations; iii) group 3, with RBD and hallucinations have been prospectively investigated at baseline and after two years throughout a clinical and neuropsychological evaluation. After two years, the group 1 continued to present normal neuropsychological tests and did not present either RBDs or hallucinations. In the group 2, the frontal impairment evidenced at baseline was confirmed; the onset of newly hallucinations was reported in a subgroup of 12 patients, who proved to be older, with a more severe executive impairment at baseline and with a more severe motor symptoms progression than those RBD patients who had not manifested hallucinations. The group 3, characterized at baseline by a more severe cognitive impairment presented, after two years, a cognitive worsening and a higher mortality rate. The longitudinal but at preliminary step investigation identified a PD subgroup of patients, in whom a common background disease profile, including the presence of RBD, could represent a "red flag" in developing further cognitive impairment. PMID- 18512750 TI - Increased predation risk modifies lizard scent-mark chemicals. AB - Variation in environmental factors plays a central role on organisms' physiological changes. However, the physiological response to predation risk has rarely been investigated in reptiles. Chemical senses are important for intraspecific communication in squamate reptiles. In male lizards Iberolacerta cyreni the maintenance of relative proportions of lipids in femoral gland secretions is costly, which may ensure honest signalling of quality. We hypothesized that increased predation risk should compromise the maintenance of such lipid proportions, as both a fear response and escaping behavior can have physiological consequences. We simulated predator attacks and found that relative proportions of lipids in femoral gland secretions changed in disturbed lizards but not in control ones. Thus, predator-prey interactions may modulate relative concentrations of chemicals in scents of lizards. Potential consequences of this effect on intraspecific chemical communication are suggested. PMID- 18512751 TI - Evaluating Parkinson's disease patients at home: utility of self-videotaping for objective motor, dyskinesia, and ON-OFF assessments. AB - The objective is to test feasibility and utility of home-based videos for assessing Parkinson's disease (PD) patients. As part of a clinical trial, patients opted between coming to the study sites or learning to videotape assessments at home. Those opting for at-home filming completed training on videotape techniques. Ten-minute films were taken at 30-minute intervals over 8.5 hours, 2 and 4 weeks after study entry using a protocol covering most items of the UDPRS motor examination and all Rush Dyskinesia Rating Scale items. After each filming, patients marked their ON/OFF status, based on prior training. We determined the number of patients who elected self-taping and the quality of video segments obtained. To assess ON/OFF patient accuracy, we compared the rater's and patient's assessment of ON/OFF at each time point. Of 12 participants, 10 elected self-videotaping and only 1 time point was missed (99.5% taping compliance). All self-recorded video segments were clear with all protocol elements included. With the exception of one missed ON/OFF rating, patient-based self-ratings occurred on time. Rating ON/OFF, UPDRS, and RDRS assessments for 8.5 hours required 170 minutes by the blinded rater. In spite of patient training, mean ON/OFF concordance between rater and patients was only 64%. At home video based self-recordings are feasible and allow accurate rater-based ON/OFF assessments. In this group of patients with no or mild fluctuations, in spite of pretrial training, patients were inaccurate in separating ON vs. OFF status. PMID- 18512752 TI - Push-and-release test predicts Parkinson fallers and nonfallers better than the pull test: comparison in OFF and ON medication states. AB - The aim of this study was to compare the Pull Test--Retropulsion Test and the Push and Release Test (P&R) as regards their ability to predict Parkinson (PD) fallers and nonfallers in relation to their medication state. Eighty-two PD patients participated in the study. Fallers (N = 36) and nonfallers (N = 46) were grouped on the basis of their fall history. Fallers were those who had fallen at least once within the last 6 months. The two groups were compared on the basis of the patients' performance in the Pull and the P&R tests, both in their "OFF" and "ON" medication state. The overall accuracy of the classification of PD patients as fallers and nonfallers was determined by means of binomial logistic regression (BLR) and the analysis of the "area under the receiver operating characteristics curve" (AUC). In the OFF medication state, the statistical analysis revealed that the Pull Test was accurate (methods BLR (AUC)) in 85.4% (0.87) of cases and the P&R Test in 86.6% (0.90). In the ON medication state, the Pull Test was only 76.8% (0.78) accurate, while the P&R Test was 87.8% (0.87) accurate. Both clinical tests are valid and relatively equivalent when assessing patients in their OFF medication state; however, the P&R Test is more accurate than the Pull Test in the ON state. This indicates that it is more widely applicable in clinical practice. PMID- 18512753 TI - Variations in gender ratios support the connection between smoking and Parkinson's disease. AB - Smokers have a decreased risk of Parkinson's disease (PD), but this association could simply be the result of a common factor causing both PD and aversion to smoking. Smoking behavior in industrialized nations has changed dramatically over the second half of the 20th century, with diverging patterns in male and female smoking rates. We therefore examined whether PD incidence changed concomitantly, as would be expected if smoking truly reduced PD risk. We performed a PUBMED search to identify relevant studies reporting male and female age-specific prevalence or incidence of PD in different countries. For each country and birth cohort we then estimated the male to female ratio in PD incidence, and correlated these ratios with the corresponding male to female ratios in smoking behavior obtained from national statistics. We show that the relative frequency of PD among women declines when the proportion of women smoking increases. The strength of this correlation (r = 0.28; P = 0.0002) is comparable to the opposite trend observed for lung cancer, and supports an overall 74% reduction in risk of PD among smokers. Our results are consistent with the hypothesis that smoking reduces the risk of PD. PMID- 18512754 TI - Young age onset of cervical dystonia. PMID- 18512755 TI - Movement disorders in Rett syndrome: an analysis of 60 patients with detected MECP2 mutation and correlation with mutation type. AB - Rett syndrome (RS) is one of the best human models to study movement disorders. Patients evolve from a hyperkinetic to a hypokinetic state, and a large series of abnormal movements may be observed along their lives such as stereotypies, tremor, chorea, myoclonus, ataxia, dystonia, and rigidity. The aim of this work was to analyze movement disorders in RS patients with a detected MECP2 mutation, as well as their correlation with genotype, in a clinically and genetically well characterized sample of patients, and thus contribute to redefine the clinical profile of this disease. In this study, we included 60 patients with detected MECP2 mutations. These were categorized and grouped for analysis, according to (1) type of change (missense or truncating, including nonsense and frameshift but also large deletions) and (2) location of the mutation. Differences were found concerning the frequency of independent gait, dystonia, type of tremor, and global score severity when comparing the group of patients with missense and truncating mutations. We also found differences in the presence, distribution, severity, or type of movement disorders in the two groups of patients according to the median duration of the disease (less than 60 months; 60 months or more). We conclude that movement disorders seem to reflect the severity and rate of progression of Rett disorder, patients with truncating mutations presenting a higher rate and more severe dystonia and rigid-akinetic syndrome, when comparing groups with similar time of disease evolution. PMID- 18512756 TI - GPi-DBS in Huntington's disease: results on motor function and cognition in a 72 year-old case. AB - Huntington's disease (HD) produces debilitating motor abnormalities that are poorly responsive to medical therapy. Deep brain stimulation (DBS) of the posteroventral globus pallidus internus (GPi) may offer a treatment option for patients with diskinetic phenotype and minimal cognitive impairment, but its role in the management of HD remains unclear and to date only two cases have been reported. We report the outcome of GPi-DBS in a 72-year-old man with HD. Stimulation at 130 Hz caused a rapid amelioration of chorea producing the worsening of bradykinesia, whereas 40 Hz stimulation (maintaining constant the total electrical energy delivered) improved chorea while preserving the ability to walk. At 1-year follow-up, chorea has completely disappeared; however, the patient was unable to stand and walk. The cognitive profile showed a progressive deterioration, with an extension of deficit from the mainly dysexecutive alterations at baseline to a more diffused cognitive deterioration. PMID- 18512757 TI - Predictors and course of health-related quality of life in Parkinson's disease. AB - We investigated how health related quality of life (HRQL) changes over time in a population-based cohort of patients with Parkinson's disease (PD), and which factors predict a lower level of HRQL in these patients. Of 227 patients with PD assessed at baseline and followed prospectively, information on HRQL-status was obtained in 111 subjects 4 years and 82 patients 8 years after inclusion. HRQL was measured by the Nottingham Health Profile (NHP). Analyses were conducted using generalized estimating equation models. The NHP total score (P < 0.001) and scores in all NHP dimensions except for sleep worsened significantly during follow-up. Steepest slope was found for the domain physical mobility (3.16, 95% CI 2.39-3.92), followed by the domains social isolation (2.22, 95% CI 1.52-2.93) and emotional reactions (1.36, 95% CI 0.74-1.97). In addition to follow-up time, higher Hoehn and Yahr staging, higher Montgomery and Aasberg Depression Rating Scale scores, and presence of insomnia at baseline were associated with lower levels of overall HRQL during follow-up. We conclude that PD has an increasing impact on HRQL as the disease progresses. During long-term follow-up, deterioration in physical mobility was the most important single factor contributing to decline in HRQL in our cohort, although distress of nonmotor character as a whole outweighed the impact of distress in physical mobility on overall HRQL. More advanced disease, higher severity of depressive symptoms, and presence of insomnia were found to be important and independent predictors of poor HRQL. (c) 2008 Movement Disorder Society. PMID- 18512758 TI - MRI detects therapeutic effects in weanling Niemann-Pick type C mice. AB - To noninvasively evaluate the early effects of Niemann-Pick type C (NPC) disease, diffusion tensor imaging (DTI) was carried out in the brains of very young (23 day-old) mice. The diffusion of water in white matter tracts of Npc1(-/-) mice at this young age was already abnormal, exhibiting decreased anisotropy, as quantified by fractional anisotropy (FA), compared with their wild-type littermates, the controls. Postmortem histological staining revealed myelin deficiencies in Npc1(-/-) mice, consistent with the reduction in FA measured in vivo. Beneficial effects of treatment with allopregnanolone and/or 2 hydroxypropyl-beta-cyclodextrin was also detectable at this age by FA, which correlated with increased myelination as seen by histology. This is the earliest detection of a therapeutic effect in Npc1(-/-) mice. PMID- 18512759 TI - N-acetylcysteine prevents beta-amyloid toxicity by a stimulatory effect on p35/cyclin-dependent kinase 5 activity in cultured cortical neurons. AB - Although previous studies have indicated that the neuroprotective effect of N acetylcysteine (NAC) required activation of the Ras-extracellular-signal regulated kinase (ERK) pathway, the detailed mechanisms and signal cascades leading to activation ERK are not clear. In the present study, we investigated the effect of NAC on A beta(25-35)-induced neuronal death. Pretreatment of neurons with NAC 1 hr before application of A beta prevented A beta-mediated cell death. NAC increased cyclin-dependent kinase 5 (Cdk5) phosphorylation, an effect that was blocked by Cdk5 inhibitor. The neuroprotective effect of NAC was significantly attenuated by Cdk5 inhibitors or in neurons transfected with Cdk5 or p35 small interfering RNA (siRNA). Conversely, pretreatment of neurons with the calpain inhibitors calpeptin or MDL28170 enhanced the neuroprotective effect of NAC. A beta(25-35) caused a significant decrease in the level of p35, with a concomitant increase in p25, which was completely prevented by NAC. This effect of NAC was blocked by the Cdk5 inhibitors roscovitine and butyrolactone. In addition, NAC increased Cdk5/p35 kinase activity but reduced Cdk5 kinase activity. A beta(25-35) treatment decreased phosphorylated levels of ERK, which could be reversed by NAC. The effect of NAC was completely blocked by Cdk5 inhibitors. NAC reversed the A beta(25-35)-induced decrease in the expression of Bcl-2, which could be blocked by the MAPK kinase (MEK) inhibitor or Cdk5 inhibitors. These results suggest that NAC-mediated neuroprotection against A beta toxicity is likely mediated by the p35/Cdk5-ERKs-Bcl-2 signal pathway. PMID- 18512760 TI - Plasticity of hippocampus following perinatal asphyxia: effects on postnatal apoptosis and neurogenesis. AB - Asphyxia during delivery produces long-term deficits in brain development, including hippocampus. We investigated hippocampal plasticity after perinatal asphyxia, measuring postnatal apoptosis and neurogenesis. Asphyxia was performed by immersing rat fetuses with uterine horns removed from ready-to-deliver rats into a water bath for 20 min. Caesarean-delivered pups were used as controls. The animals were euthanized 1 week or 1 month after birth. Apoptotic nuclear morphology and DNA breaks were assessed by Hoechst and TUNEL assays. Neurogenesis was estimated by bromodeoxyuridine/MAP-2 immunocytochemistry, and the levels and expression of proteins related to apoptosis and cell proliferation were measured by Western blots and in situ hybridization, respectively. There was an increase of apoptosis in CA1, CA3, and dentate gyrus (DG) and cell proliferation and neurogenesis in CA1, DG, and hilus regions of hippocampus 1 week after asphyxia. The increase of apoptosis in CA3 and cell proliferation in the suprapyramidal band of DG was still observed 1 month following asphyxia. There was an increase of BAD, BCL-2, ERK2, and bFGF levels in whole hippocampus and bFGF expression in CA1 and CA2 and hilus at P7 and P30. There was a concomitant decrease of phosphorylated-BAD (Ser112) levels. The increase of BAD levels supports the idea of delayed cell death after perinatal asphyxia, whereas the increases of BCL-2, ERK2, and bFGF levels suggest the activation of neuroprotective and repair pathways. In conclusion, perinatal asphyxia induces short- and long-term regionally specific plastic changes, including delayed cell death and neurogenesis, involving pro- and antiapoptotic as well as mitogenic proteins, favoring hippocampal functional recovery. PMID- 18512761 TI - Neuroprotection of gamma-aminobutyric acid receptor agonists via enhancing neuronal nitric oxide synthase (Ser847) phosphorylation through increased neuronal nitric oxide synthase and PSD95 interaction and inhibited protein phosphatase activity in cerebral ischemia. AB - It is well documented that exitotoxicity induced by N-methyl-D-aspartate (NMDA) receptor activation plays a pivotal role in delayed neuronal death in the hippocampal CA1 region after transient global ischemia. However, the effect of gamma-aminobutyric acid (GABA) receptor activation is uncertain in ischemia brain injury. The aim of this study was to investigate whether the enhancement of GABA receptor activity could inhibit NMDA receptor-mediated nitric oxide (NO) production by neuronal NO synthase (nNOS) in brain ischemic injury. The results showed that both the GABA(A) receptor agonist muscimol and the GABA(B) receptor agonist baclofen had neuroprotective effect, and the combination of two agonists could significantly protect neurons against death induced by ischemia/reperfusion. Coapplication of muscimol with baclofen not only enhanced nNOS (Ser847) phosphorylation but also increased the interaction of nNOS with PSD95 at 6 hr and 1 day of reperfusion. Interestingly, the inhibitors of calcineurin and PP1/PP2A could enhance nNOS phosphorylation at Ser847 site at 1 day of reperfusion after ischemia but not at 6 hr of reperfusion. From these data, we conclude that GABA receptor activation could exert its neuroprotective effect through increasing nNOS (Ser847) phosphorylation by different mechanisms at 6 hr and 1 day of reperfusion. The increased interaction of nNOS and postsynaptic density-95 induced by GABA agonists is responsible for nNOS (Ser847) phosphorylation at both time points, but at 1 day of reperfusion the inhibition of protein phosphatase activity by GABA agonists also contributes to the neuroprotection. Our results suggest that GABA receptor agonists may serve as a potential and important neuroprotectant in therapy for ischemic stroke. PMID- 18512762 TI - Hoxa1 is required for the retinoic acid-induced differentiation of embryonic stem cells into neurons. AB - The ability of embryonic stem (ES) cells to differentiate into different cell fates has been extensively evaluated, and several protocols exist for the generation of various types of cells from mouse and human ES cells. We used a differentiation protocol that involves embryoid body formation and all-trans retinoic acid (RA, 5 microM) treatment (EB/5 microM RA) to test the ability of Hoxa1 null ES cells to adopt a neuronal fate. Hoxa1(-/-) ES cells, when treated in this EB/5 microM RA protocol, failed to differentiate along a neural lineage; Hoxa1(-/-) ES cells express severalfold lower levels of many neuronal differentiation markers, including nestin, beta-tubulin III, and MAP2, and conversely, higher levels of endodermal differentiation markers (i.e., Sox17, Col4a1) than wild type (Wt) cells. Reintroduction of exogenous Hoxa1, under the control of the metallothionein I promoter, into Hoxa1(-/-) ES cells restored their capacity to generate neurons. Moreover, overexpression of Sox17, a gene that regulates endodermal differentiation, in Wt ES cells resulted in endodermal differentiation and in a complete abolition of beta-tubulin III expression. Thus, Hoxa1 activity is essential for the neuronal differentiation of ES cells in the presence of all-trans-RA, and Hoxa1 may promote neural differentiation by inhibiting Sox17 expression. Pharmacological manipulation of Hoxa1 levels may provide a method for promoting neuronal differentiation for therapeutic uses. Furthermore, because mutations in the Hoxa1 gene can cause autism spectrum disorder in humans, these data also provide important mechanistic insights into the early developmental processes that may result in this disorder. PMID- 18512763 TI - Protective effects of erythropoietin on tau phosphorylation induced by beta amyloid. AB - Neuropathological studies have demonstrated that the presence of neurofibrillary tangles (NFTs) is one of the most prominent pathologic characteristics of Alzheimer's disease (AD). The microtubule-associated protein tau is the major component of NFTs, and its abnormal hyperphosphorylation leads to the destabilization of microtubules, impaired axonal transport, and eventual death of the neurons. The hematopoietic cytokine erythropoietin (Epo) is now considered as a viable agent with regard to central nervous system injury in a variety of cellular systems. Here we report that Epo prevented tau hyperphosphorylation in SH-SY5Y cells exposed to the beta-amyloid peptide and that this effect may depend on the PI3K/Akt-GSK-3beta pathway. This study provides new molecular insight into the neuroprotective effect of Epo and suggests its possible therapeutic role in the management of AD. PMID- 18512764 TI - Pure red cell aplasia in a patient with adult-onset Still's disease. PMID- 18512765 TI - Cold agglutinin-associated hemolytic anemia due to brucellosis: first case report. PMID- 18512766 TI - Promoter hypermethylation-mediated down-regulation of CXCL12 in human astrocytoma. AB - It has recently been demonstrated that CXCL12 is absent in colonic carcinoma, and hypermethylation of CXCL12 contributes to CXCL12/CXCR4 signaling in carcinoma metastasis. However, the role of CXCL12/CXCR4 axis, especially CXCL12, in the regulation of tumor invasiveness is largely still unknown. Using real-time quantitative RT-PCR assays, we observed that CXCR4 expression increased with increasing WHO grade in astrocytoma, suggesting that CXCR4 may be a marker of aggressive biological behavior of astrocytoma. Methylation of CXCL12 was detected in 34.2% (26/76) of astrocytomas by methylation-specific PCR. Epigenetic inactivation of CXCL12 was implicated mainly in low-grade astrocytomas, via DNA hypermethylation by DNMT1, -3A, and -3B; 21.1% (16/76) of the astrocytomas showed reduced or lack of CXCL12 expression, in line with epigenetic silencing of gene transcripts. However, it is interesting to note that 61.8% (47/76) of tumors, mainly high-grade astrocytomas, displayed elevated transcription of CXCL12. The expression levels of CXCL12 mRNA in glioblastomas (WHO grade IV) were significantly higher than in normal brain tissues. In summary, our data show that CXCL12 promoter hypermethylation is an early event in astrocytoma development. However, the high expressions of CXCR4 and CXCL12 in glioblastomas, the more invasive astrocytomas, suggest a different role of CXCL12/CXCR4 signaling axis in astrocytoma progression. PMID- 18512767 TI - The relationship between CAG repeat length and clinical progression in Huntington's disease. AB - The objective of this study was to examine the relationship between CAG repeat length (CAGn) and clinical progression in patients with Huntington's disease (HD). There are conflicting reports about the relationship between CAGn and clinical progression of HD. We conducted an analysis of data from the Coenzyme Q10 and Remacemide Evaluation in Huntington's Disease (CARE-HD) clinical trial. We modeled progression over 30 months on the Unified Huntington's Disease Rating Scale (UHDRS) and supplemental neuropsychological and behavioral tests using multiple linear regression. Mean subject age was 47.9 +/- 10.5 years and mean CAGn was 45.0 +/- 4.1. Multiple linear regression revealed statistically significant associations between CAGn and worsening on several motor, cognitive, and functional outcomes, but not behavioral outcomes. Many effects were clinically important; 10 additional CAG repeats were associated with an 81% increase in progression on the Independence Scale. These associations were not observed in the absence of age adjustment. Age at the time of assessment confounds the association between CAGn and progression. Adjusting for age shows that longer CAGn is associated with greater clinical progression of HD. This finding may account for the variable results from previous studies examining CAGn and progression. Adjusting for CAGn may be important for clinical trials. PMID- 18512768 TI - Comment on "individualized assessment of quality of life in idiopathic Parkinson's disease". PMID- 18512771 TI - Lymphoid follicles in joints: what do they mean? PMID- 18512772 TI - Anti-CD20 therapy in patients with rheumatoid arthritis: predictors of response and B cell subset regeneration after repeated treatment. AB - OBJECTIVE: B cell depletion with the anti-CD20 antibody rituximab has proven efficacy in patients with rheumatoid arthritis (RA). The effects on B cell homeostasis after repeated treatments and the relationship of certain B cell subsets to clinical response or relapse are currently not known. METHODS: In this open-label study, 17 patients with RA refractory to standard therapy were treated with 1 cycle of rituximab. Of these 17 patients, 11 received a second cycle of rituximab therapy. Immunophenotyping was performed before therapy and during B cell recovery. RESULTS: Twelve of 17 patients showed a good European League Against Rheumatism response after receiving 1 cycle of rituximab therapy. At the time of B cell recovery, the IgD+,CD27+ memory B cell subset was significantly larger (P = 0.019) in the nonresponder group. Within the group of 12 responders, 6 patients, whose disease was characterized by a significantly higher proportion of overall CD27+ memory B cells before therapy, experienced an early relapse (weeks 24-40 posttreatment). Eleven patients were re-treated, again resulting in a good clinical response. B cell reconstitution followed a similar pattern after each cycle. The early reconstitution phase was characterized by immature CD38++,IgD+,CD10+ B cells, whereas the number of naive B cells increased continuously thereafter. The number of memory B cells was still reduced at the time of the second depletion but recovered to levels similar to those following the first cycle of therapy. CONCLUSION: Data derived from repeated B lymphocyte depletion with rituximab in patients with RA suggest that analysis of certain memory B cell subsets provides information on efficacy, response, and late as well as early relapse, consistent with the conclusion that targeting memory B cells is a key to its mechanism of action. PMID- 18512773 TI - Patients with pulmonary tuberculosis are frequently positive for anti-cyclic citrullinated peptide antibodies, but their sera also react with unmodified arginine-containing peptide. AB - OBJECTIVE: The anti-cyclic citrullinated peptide (anti-CCP) enzyme-linked immunosorbent assay (ELISA) has high sensitivity and specificity for rheumatoid arthritis (RA). However, detection of anti-CCP in patients with active pulmonary tuberculosis (TB) has recently been reported. To determine whether this activity was specific for the citrullinated residue, the specificity of anti-CCP-positive sera for CCP versus that for unmodified arginine-containing peptide (CAP) was examined in patients with TB and compared with that in patients with RA. METHODS: Anti-CCP and anti-CAP in sera from patients with pulmonary TB (n = 49), RA patients (n = 36), and controls (n = 18) were tested by ELISA. Sera were available at diagnosis from most TB patients. All TB patients were treated with a combination of 2-4 antibiotics for at least 6 months, and sera were collected over time. RESULTS: Anti-CCP was found in 37% of TB patients and in 43% of RA patients. CAP reactivity was more common in TB than in RA. High anti-CCP:anti-CAP ratios (>2.0) were seen far more commonly in anti-CCP-positive RA patients than in anti-CCP-positive TB patients (94% versus 22%). Anti-CCP was inhibited by CCP peptide in sera from RA patients, but not in sera from TB patients. A slight increase in anti-CCP was common after initiating treatment for TB, although the anti-CCP level decreased after 1-2 months. CONCLUSION: Anti-CCP is frequently present in patients with active TB. However, many anti-CCP-positive TB sera also reacted with CAP, and anti-CCP:anti-CAP ratios in TB sera were low. Anti-CCP:anti CAP ratios should be useful clinically for distinguishing CCP-specific reactivity seen in RA from reactivity with both CCP and CAP frequently seen in pulmonary TB. PMID- 18512774 TI - Synovial lymphoid neogenesis does not define a specific clinical rheumatoid arthritis phenotype. AB - OBJECTIVE: To investigate the relationship between lymphoid neogenesis in the synovium of patients with rheumatoid arthritis (RA) and characteristics of inflammation and disease severity. METHODS: Arthroscopic synovial biopsy was performed in 103 patients with active RA (Disease Activity Score 28-joint assessment >or=3.2) who had not received treatment with biologic agents. Sections were stained and assessed by digital image analysis. Lymphocyte aggregates were counted and graded for size (1-3). Synovial lymphoid neogenesis was defined as the presence of grade 2 or 3 aggregates and subclassified based on the presence of follicular dendritic cells (FDCs). RESULTS: Lymphoid neogenesis was present in 31% of the RA synovial tissues, whereas an additional 25% contained only grade 1 aggregates. FDCs were present in 28% of the samples with lymphoid neogenesis, corresponding to 8% of the total RA cohort. Histologically, synovia with lymphoid neogenesis showed increased infiltration by T and B lymphocytes, plasma cells, and macrophages, and increased expression of tumor necrosis factor alpha and lymphotoxin beta compared with samples without lymphoid neogenesis. Patients with lymphoid neogenesis also had higher C-reactive protein levels, erythrocyte sedimentation rates, and leukocyte and thrombocyte counts, but exhibited no increase in the severity of clinical signs and symptoms. Of importance, there was no relationship between the presence of lymphoid neogenesis and IgM rheumatoid factor or anti-citrullinated protein antibodies. The presence of lymphocyte aggregates with FDCs did not define a specific clinical phenotype compared with lymphocyte aggregates without FDCs. CONCLUSION: These findings indicate that synovial lymphoid neogenesis is associated with more severe synovial and systemic inflammation, but this is not confined to a specific clinical subset of RA. PMID- 18512775 TI - Dramatic regulation of heparanase activity and angiogenesis gene expression in synovium from patients with rheumatoid arthritis. AB - OBJECTIVE: Although heparanase is recognized as a proangiogenic factor, the involvement of heparanase in rheumatoid arthritis (RA) is unclear. In this study, we assessed heparanase activity in synovial fluid (SF) and synovial tissue (ST) from patients with RA or osteoarthritis (OA), and analyzed the expression of angiogenic pathway-focused genes in ST from RA and OA patients. METHODS: SF and ST were obtained from the knees of patients with either RA or OA and from asymptomatic donors with no documented history of degenerative or inflammatory joint diseases. Heparanase activity was determined by an enzymatic assay using a radiolabeled substrate, and the presence of heparanase in ST was demonstrated by Western blotting. The expression of angiogenesis genes, including heparanase, in ST was analyzed by real-time quantitative polymerase chain reaction. RESULTS: Heparanase activity was dramatically higher (>100-fold) in SF and ST from RA patients than in SF and ST from OA patients and asymptomatic donors. Active heparanase enzyme was detected and heparanase messenger RNA was up-regulated in ST from RA patients. We also found that angiogenesis gene expression was significantly regulated in RA synovium, and was correlated with heparanase activity. CONCLUSION: These findings are novel and contribute to our understanding of joint destruction in RA, suggesting that heparanase may be a reliable prognostic factor for RA progression and an attractive target for the treatment of RA. PMID- 18512776 TI - Decreased lubricin concentrations and markers of joint inflammation in the synovial fluid of patients with anterior cruciate ligament injury. AB - OBJECTIVE: To study the effect of anterior cruciate ligament (ACL) injury on lubricin concentrations in synovial fluid (SF) and its correlation with time postinjury, inflammatory cytokines, lubricin-degrading enzymes, and SF proteoglycan content. METHODS: SF samples were obtained from both knees of 30 patients with unilateral ACL insufficiency, 32-364 days postinjury. Lubricin, inflammatory cytokines (interleukin-1beta [IL-1beta], tumor necrosis factor alpha [TNFalpha], and IL-6), and catabolic enzymes (procathepsin B and neutrophil elastase) were measured in SF from injured and contralateral (uninjured) joints, by enzyme-linked immunosorbent assay. Sulfated glycosaminoglycan (sGAG) levels in the SF were measured by Alcian blue binding assay. RESULTS: SF lubricin concentrations were significantly (P < 0.001) reduced at an early stage following ACL injury when compared with those in the contralateral joint. Within 12 months, the lubricin concentration in the injured knee (slope = 0.006, SE = 0.00010, P < 0.001) approached that in the contralateral knee, which did not change with time (slope = -0.0002, SE = 0.00050, P = 0.71). TNFalpha levels showed a significant negative relationship with log2 lubricin levels. IL-1beta, TNFalpha, IL-6, procathepsin B, and neutrophil elastase concentrations in SF from injured knees were greater in samples from recently injured knees compared with those that were chronically injured. There were no detectable cytokines or enzymes in the SF of contralateral joints. Concentrations of sGAG were significantly (P = 0.0002) higher in the SF from injured knees compared with the contralateral joints. CONCLUSION: The decrease in SF lubricin concentrations following ACL injury may place the joint at an increased risk of wear-induced damage as a consequence of lack of boundary lubrication, potentially leading to secondary osteoarthritis. The decrease in SF lubricin was associated with an increase in levels of inflammatory cytokines. PMID- 18512777 TI - Relationship of meniscal damage, meniscal extrusion, malalignment, and joint laxity to subsequent cartilage loss in osteoarthritic knees. AB - OBJECTIVE: Progressive knee osteoarthritis (OA) is believed to result from local factors acting in a systemic environment. Previous studies have not examined these factors concomitantly or compared quantitative and qualitative cartilage loss outcomes. The aim of this study was to test whether meniscal damage, meniscal extrusion, malalignment, and laxity each predicted tibiofemoral cartilage loss after controlling for the other factors. METHODS: Laxity and alignment were measured at baseline in individuals with knee OA. Magnetic resonance imaging included spin-echo coronal and sagittal imaging for meniscal scoring and axial and coronal spoiled gradient echo sequences with water excitation for cartilage quantification. Tibial and weight-bearing femoral condylar subchondral bone area and cartilage surface were segmented. Cartilage volume, denuded bone area, and cartilage thickness were quantified in each plate, with progression defined as cartilage loss >2 times the coefficient of variation for each plate. Qualitative outcome was assessed as worsening of the cartilage score. Logistic regression analysis with generalized estimating equations yielded odds ratios for each factor, adjusting for age, sex, body mass index, and the other factors. RESULTS: We studied 251 knees in 153 persons. After full adjustment, medial meniscal damage predicted medial tibial cartilage volume loss and tibial and femoral denuded bone increase, while varus malalignment predicted medial tibial cartilage volume and thickness loss and tibial and femoral denuded bone increase. Lateral meniscal damage predicted every lateral outcome. Laxity and meniscal extrusion had inconsistent effects. After full adjustment, no factor except medial laxity predicted qualitative outcome. CONCLUSION: Using quantitative cartilage loss assessment, local factors that independently predicted tibial and femoral loss included medial meniscal damage and varus malalignment (medially) and lateral meniscal damage (laterally). A measurement of quantitative outcome was more sensitive at revealing these relationships than a qualitative approach. PMID- 18512778 TI - Association between findings on delayed gadolinium-enhanced magnetic resonance imaging of cartilage and future knee osteoarthritis. AB - OBJECTIVE: To examine the predictive value of the delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) index with regard to future radiographic osteoarthritis (OA). METHODS: In 1998, 17 knees in 11 men and 4 women with knee pain, normal results of weight-bearing radiography, and arthroscopic cartilage changes ranging from superficial fibrillation to fissuring and softening were examined using dGEMRIC. Six years later, 16 of the 17 knees were reassessed for radiographic OA changes. RESULTS: At followup, 9 of the 16 knees showed radiographic OA changes. Two of them had undergone a knee joint replacement due to OA. In the knees with radiographic OA, the dGEMRIC index at baseline was lower than that in the knees without radiographic OA (P = 0.03). CONCLUSION: The results of the present study support the dGEMRIC index as a clinically relevant measure of cartilage integrity and suggest that a low index may be predictive of the development of knee OA. PMID- 18512779 TI - Synovial fluid mesenchymal stem cells in health and early osteoarthritis: detection and functional evaluation at the single-cell level. AB - OBJECTIVE: Arthritic synovial fluid (SF) contains mesenchymal stem cells (MSCs), which could simply reflect their shedding from diseased joint structures. This study used the bovine model to explore SF MSCs in health and enumerated them at the earliest stages of human osteoarthritis (OA) in radiographically normal joints. METHODS: Clonogenicity and multipotentiality of normal bovine SF MSCs were compared with donor-matched bone marrow (BM) MSCs at the single-cell level. The colony-forming unit-fibroblastic assay was used for MSC enumeration. The XTT assay was employed to assess cell proliferation, and flow cytometry was used to investigate the marker phenotype of bovine and human SF MSCs. RESULTS: Single MSCs were present in normal bovine SF, and 96% of them were able to expand at least 1 million-fold. These cells were CD271-, multipotential, considerably more clonogenic, and less adipogenic than matched BM MSCs. In both pellet assays and on polyglycolic acid scaffolds, SF clones displayed consistent chondrogenic differentiation, while BM clones were variable. MSCs were present in arthroscopically normal human joints and were increased 7-fold in early OA (P = 0.034). Their numbers correlated with numbers of free microscopic synovial tissue fragments (r = 0.826, P < 0.0001). OA SF had a growth-promoting effect on synovial MSCs. CONCLUSION: This study confirms the presence of MSCs in normal SF and shows their numerical increase in early human OA. SF MSCs are likely to originate from synovium. These findings provide a platform for the exploration of the potential role of SF MSCs in joint homeostasis and for investigation of their utility in novel joint regeneration strategies. PMID- 18512780 TI - Different T cell subsets in the nodule and synovial membrane: absence of interleukin-17A in rheumatoid nodules. AB - OBJECTIVE: To determine gene expression of the interleukin-17 (IL-17) family members (IL-17A-F) in rheumatoid subcutaneous nodules, and to assess the cytokines involved in regulating IL-17A expression. METHODS: Total RNA was isolated from 19 nodules obtained from 16 different patients with rheumatoid arthritis (RA). Reverse transcription-polymerase chain reaction (PCR) was used to screen for gene expression of the IL-17 subtypes (IL-17A-F) in all nodules. Quantitative real-time PCR was used to measure the expression of interferon-gamma (IFN gamma), IL-6, IL-23, IL-12, and transforming growth factor beta (TGFbeta), relative to GAPDH as control, in a subset of 10 nodules. RESULTS: IL-17A gene expression was present in only 1 of 19 nodules, IL-17B in 17 of 19 nodules, IL 17C in 18 of 19 nodules, IL-17D in 16 of 19 nodules, and IL-17E in 3 of 19 nodules. IL-17F was absent in all samples. Cytokines that stimulate IL-17A production (IL-6, IL-23) as well as those that inhibit IL-17A production (IL-12, IFN gamma, TGFbeta) were present in the majority of nodules. Quantitative real time PCR showed a similar pattern of gene expression for the individual cytokines between the different nodules. The mean +/- SD expression of IL-6 relative to GAPDH was 2.28 +/- 2.2 ng, and that of TGFbeta was 2.96 +/- 1.14 ng. There was a lower relative expression of IL-23 (0.05 +/- 0.05 ng), while the expression of IFN gamma was 0.67 +/- 0.68 ng and that of IL-12 was 0.48 +/- 0.23 ng. CONCLUSION: IL-17 family members are varyingly expressed in rheumatoid nodules. The paucity of IL-17A in nodules suggests an important difference from that observed in the synovium. The expression of IL-23 below a critical threshold level seems the most likely explanation for the virtual absence of IL-17A. The presence of tissue destruction within the nodule despite the absence of IL-17A suggests that IL-17A may be an important amplifier rather than an absolute requirement for inflammation in RA. PMID- 18512781 TI - Suppressor activity among CD4+,CD25++ T cells is discriminated by membrane-bound tumor necrosis factor alpha. AB - OBJECTIVE: Previous studies have shown that the suppressive capacity of CD4+,CD25++ T cells is compromised in patients with rheumatoid arthritis (RA) and restored by anti-tumor necrosis factor alpha (anti-TNFalpha) therapy. Given the lack of specific cell surface markers for human Treg cells, this study aimed to define surface markers for identifying and enriching Treg cells with enhanced regulatory ability within the CD4+,CD25++ T cell compartment and to provide additional understanding of the effects of anti-TNFalpha antibodies in humans. METHODS: The expression of membrane-bound TNFalpha in human peripheral blood CD4+ T cells was analyzed by flow cytometry in healthy individuals and RA patients before and after anti-TNFalpha treatment. Membrane-bound TNFalpha-positive and TNFalpha-negative CD4+,CD25++ T cells were purified by fluorescence-activated cell sorting, and their suppressive capacity was assessed in vitro by a standard suppression assay. RESULTS: A substantial number of CD4+,CD25++ T cells expressed membrane-bound TNFalpha. Membrane-bound TNFalpha-positive CD4+,CD25++ T cells displayed reduced antiinflammatory cytokine production and less potent suppressor capacity, since 4 times more cells were required to achieve 50% inhibition compared with their membrane-bound TNFalpha-negative counterparts. Treatment of RA patients with TNFalpha-specific antibodies led to a reduction in the number of membrane-bound TNFalpha-positive CD4+,CD25++ T cells from peripheral blood. CONCLUSION: Our data indicate that the absence of membrane-bound TNFalpha on CD4+,CD25++ T cells can be used to characterize and enrich for Treg cells with maximal suppressor potency. Enrichment of membrane-bound TNFalpha-negative CD4+,CD25+ cells in the CD4+,CD25++ T cell compartment may contribute to restoring the compromised suppressive ability of CD4+,CD25++ T cell populations in RA patients after anti-TNFalpha treatment. PMID- 18512782 TI - Cytokines secreted in response to Toll-like receptor ligand stimulation modulate differentiation of human Th17 cells. AB - OBJECTIVE: Th17 cells (interleukin-17 [IL-17]-secreting T helper cells) have been implicated in the pathogenesis of rheumatoid arthritis and other autoimmune diseases, but the soluble factors that influence human Th17 differentiation have yet to be fully elucidated. This study was undertaken to investigate the hypothesis that the cytokines secreted by human peripheral blood mononuclear cells (PBMCs) in response to a subset of Toll-like receptor (TLR) ligands would influence Th17 polarization. METHODS: Supernatants from human PBMCs treated with a panel of TLR agonists were tested for their ability to induce de novo IL-17 production in naive T helper cells. Multiplex cytokine analysis was used to identify candidate cytokines for subsequent blocking and sufficiency experiments. RESULTS: Conditioned media from PBMCs stimulated with TLR-4 or TLR-8/7 agonists, but not from those stimulated with TLR-2/1, -3, or -9 agonists, evoked robust secretion of IL-17 by T helper cells, independent of coculture with antigen presenting cells. Multiplex analysis of 22 cytokines and chemokines identified a 6-factor cytokine signature that significantly correlated with IL-17-inducing activity. T cell activation in the presence of recombinant IL-1beta, IL-6, and IL 23 reconstituted robust IL-17 production, and this was enhanced by transforming growth factor beta (TGFbeta). IL-6 suppressed the expression of forkhead box P3 and reversed TGFbeta-mediated inhibition of T cell proliferation, but did not trigger IL-17 secretion. IL-17 production was completely abrogated by anti-IL-1 or IL-1 receptor antagonist and partially inhibited by anti-IL-6, anti-IL-2, or exogenous retinoic acid, but not by anti-tumor necrosis factor alpha. IL-1beta and IL-6 independently induced IL-21 secretion, but the presence of IL-21 alone was not sufficient for IL-17 production. CONCLUSION: These results indicate that ligation of a subset of TLRs generates proinflammatory cytokines that combine to potentiate human Th17 differentiation. PMID- 18512783 TI - The rheumatoid arthritis-associated allele HLA-DR10 (DRB1*1001) shares part of its repertoire with HLA-DR1 (DRB1*0101) and HLA-DR4 (DRB*0401). AB - OBJECTIVE: To identify the peptide anchor motif for the rheumatoid arthritis (RA) related HLA allele, DR10, and find shared natural ligands or sequence similarities with the other disease-associated alleles, DR1 and DR4. METHODS: The HLA-DR10-associated peptides were purified, and a proportion of these natural ligands were de novo sequenced by mass spectrometry. Based on crystallographic structures, the complexes formed by peptide influenza virus hemagglutinin HA306 318 with DR1, DR4, and DR10 were modeled, and binding scores were obtained. RESULTS: A total of 238 peptides were sequenced, and the anchor motif of the HLA DR10 peptide repertoire was defined. A large proportion of the DR10-associated peptides had the structural features to bind DR1 and DR4 but were theoretical nonbinders to the negatively associated alleles DR15 and DR7. Among the sequenced ligands, 10 had been reported as ligands to other RA-associated alleles. Modeling data showed that peptide HA306-318 can bind DR1, DR4, and DR10 with similar affinities. CONCLUSION: The data show the presence of common peptides in the repertoires of RA-associated HLA alleles. The combination of the shared epitope present in DR1, DR4, and DR10 together with common putative arthritogenic peptide(s) could influence disease onset or outcome. PMID- 18512784 TI - Gene expression analysis of macrophages derived from ankylosing spondylitis patients reveals interferon-gamma dysregulation. AB - OBJECTIVE: To determine whether macrophages, a type of cell implicated in the pathogenesis of ankylosing spondylitis (AS), exhibit a characteristic gene expression pattern. METHODS: Macrophages were derived from the peripheral blood of 8 AS patients (median disease duration 13 years [range <1-43 years]) and 9 healthy control subjects over 7 days with the use of granulocyte-macrophage colony-stimulating factor. Cells were stimulated for 24 hours with interferon gamma (IFN gamma; 100 units/ml), were left untreated for 24 hours, or were treated for 3 hours with lipopolysaccharide (LPS; 10 ng/ml). RNA was isolated and examined by microarray and real-time quantitative reverse transcription polymerase chain reaction analysis. RESULTS: Microarray analysis revealed 198 probe sets detecting the differential expression of 141 unique genes in untreated macrophages from AS patients compared with healthy controls. Clustering and principal components analysis clearly distinguished AS patients and controls. Of the differentially expressed genes, 78 (55%) were IFN-regulated, and their relative expression indicated a "reverse" IFN signature in AS patient macrophages, where IFN gamma-up-regulated genes were underexpressed and down regulated genes were overexpressed. Treatment of macrophages with exogenous IFN gamma normalized the expression of these genes between patients and controls. In addition, the messenger RNA encoded by the IFN gamma gene was approximately 2 fold lower in AS patient macrophages at baseline (P = 0.004) and was poorly responsive to LPS (P = 0.018), as compared with healthy controls. CONCLUSIONS: Our findings reveal consistent differences in gene expression in macrophages from AS patients, with evidence of a striking "reverse" IFN signature. Together with poor expression and responsiveness of the IFN gamma gene, these results suggest that there may be a relative defect in IFN gamma gene regulation, with autocrine consequences and implications for disease pathogenesis. PMID- 18512785 TI - Amelioration of collagen-induced arthritis in rats by adenovirus-mediated PTEN gene transfer. AB - OBJECTIVE: The phosphatidylinositol 3-kinase (PI 3-kinase)/Akt pathway is known to be activated in rheumatoid arthritis (RA) synovial tissue, which impacts cell growth, proliferation, survival, and migration. Phosphatase and tensin homolog deleted from chromosome 10 (PTEN) functions as a negative regulator of PI 3 kinase signaling, thus blocking Akt activation. The aim of this study was to examine the effect of PTEN gene transfer in rats with collagen-induced arthritis (CIA). METHODS: Adenoviral vectors encoding human PTEN (AdPTEN) or beta galactosidase (AdLacZ) were injected intraarticularly into rats with CIA, and their treatment responses were monitored by measures of clinical, radiographic, and histologic changes. The expression of phosphorylated Akt, total Akt, vascular endothelial growth factor (VEGF), proinflammatory cytokines, and chemokines, as well as the extent of microvessel density in the ankle joints were determined. RESULTS: AdPTEN treatment reduced Akt phosphorylation and decreased VEGF production in human RA synovial fibroblasts. Compared with AdLacZ treatment of the rats with CIA, AdPTEN treatment significantly reduced ankle circumference, articular index scores, radiography scores, and histology scores, and also decreased microvessel density and levels of VEGF and interleukin-1beta. Furthermore, PTEN gene transfer led to down-regulation of Akt activation and increased apoptosis in the ankle joints. CONCLUSION: This study is the first to demonstrate the in vivo effect of intraarticular gene delivery of PTEN on amelioration of arthritis symptoms in rats with CIA, which involved antiangiogenic, antiproliferative, and antiinflammatory effects of PTEN via inhibition of the PI 3-kinase/Akt signaling pathway. Our findings also implicate the PI 3-kinase/Akt pathway as a therapeutic target for the treatment of RA or other inflammatory diseases. PMID- 18512786 TI - Ciprofloxacin-mediated cell proliferation inhibition and G2/M cell cycle arrest in rat tendon cells. AB - OBJECTIVE: To investigate the effect of ciprofloxacin on the proliferation and cell cycle progression of tendon cells, and to explore the potential molecular mechanism of ciprofloxacin-associated tendinopathy by analyzing the expression of cell cycle-related cyclin and cyclin-dependent kinase (CDK). METHODS: Rat Achilles tendon cells were treated with ciprofloxacin and then assessed by MTT assay, flow cytometric analysis, and fluorescence confocal microscopy. Levels of messenger RNA (mRNA) for CDK-1 and cyclin B were determined by reverse transcriptase-polymerase chain reaction. Protein expression of CDK-1, cyclin B, checkpoint kinase 1 (CHK-1), and polo-like kinase 1 (PLK-1) was determined by Western blot analysis. RESULTS: Ciprofloxacin inhibited tendon cell proliferation and caused cell cycle arrest at the G2/M phase. Confocal microscopy revealed that chromosomes in ciprofloxacin-treated cells neither properly aligned along the equatorial planes nor segregated successfully during metaphase. Mitotic arrest, misaligned chromosomes, and poor bipolar spindle formation were observed in ciprofloxacin-treated cells. CDK-1 and cyclin B protein and mRNA were both down regulated. CHK-1 protein expression was also suppressed, but PLK-1 protein expression was up-regulated by ciprofloxacin. CONCLUSION: Our findings suggest a possible mechanism of ciprofloxacin-associated tendinopathy. Down-regulation of CHK-1 and up-regulation of PLK-1 may account for mitotic arrest observed in ciprofloxacin-treated cells. PMID- 18512787 TI - Evidence of a novel aggrecan-degrading activity in cartilage: Studies of mice deficient in both ADAMTS-4 and ADAMTS-5. AB - OBJECTIVE: To characterize aggrecan catabolism and the overall phenotype in mice deficient in both ADAMTS-4 and ADAMTS-5 (TS-4/TS-5 Delta-cat) activity. METHODS: Femoral head cartilage from the joints of TS-4/TS-5 Delta-cat mice and wild-type mice were cultured in vitro, and aggrecan catabolism was stimulated with either interleukin-1alpha (IL-1alpha) or retinoic acid. Total aggrecan release was measured, and aggrecanase activity was examined by Western blotting using neoepitope antibodies for detecting cleavage at EGE 373-374 ALG, SELE 1279-1280 GRG, FREEE 1467-1468 GLG, and AQE 1572-1573 AGEG. Aggrecan catabolism in vivo was examined by Western blotting of cartilage that had been extracted immediately ex vivo. RESULTS: TS-4/TS-5 Delta-cat mice were viable, fertile, and phenotypically normal. TS-4/TS-5 Delta-cat cartilage explants did not release aggrecan in response to IL-1alpha, and there was no detectable increase in aggrecanase neoepitopes. TS-4/TS-5 Delta-cat cartilage explants released aggrecan in response to retinoic acid. There was no retinoic acid-stimulated cleavage at either EGE 373-374 ALG or AQE 1572-1573 AGEG. There was a low level of cleavage at SELE 1279 1280 GRG and major cleavage at FREEE 1467-1468 GLG. Ex vivo, cleavage at FREEE 1467-1468 GLG was substantially reduced, but still present, in TS-4/TS-5 Delta cat mouse cartilage compared with wild-type mouse cartilage. CONCLUSION: An aggrecanase other than ADAMTS-4 and ADAMTS-5 is expressed in mouse cartilage and is up-regulated by retinoic acid but not IL-1alpha. The novel aggrecanase appears to have different substrate specificity from either ADAMTS-4 or ADAMTS-5, cleaving E-G bonds but not E-A bonds. Neither ADAMTS-4 nor ADAMTS-5 is required for normal skeletal development or aggrecan turnover in cartilage. PMID- 18512788 TI - Glucocorticoid excess in mice results in early activation of osteoclastogenesis and adipogenesis and prolonged suppression of osteogenesis: a longitudinal study of gene expression in bone tissue from glucocorticoid-treated mice. AB - OBJECTIVE: Glucocorticoid (GC) excess induces alterations in bone metabolism that weaken bone structure and increase fracture risk. The aim of this study was to identify genes associated with bone metabolism in GC-treated mice, by performing a microarray analysis. METHODS: Long bones from mice exposed to GC excess were collected after 0, 7, 28, and 56 days of treatment, to measure bone microarchitecture and extract RNA for microarray analyses. RESULTS: Bone loss in this animal model was confirmed by changes in bone turnover markers as well as bone architecture, as measured by microfocal computed tomography. GC excess induced an early up-regulation of genes involved in osteoclast activation, function, and adipogenesis, which peaked on day 7. The expression of genes associated with osteoclast cytoskeletal reorganization and genes associated with matrix degradation peaked on day 28. On day 28 and day 56, the expression of genes associated with osteoblast activation and maturation was decreased from baseline, while the expression of Wnt antagonists was increased. In addition, the expression of genes expressed in osteocytes associated with bone mineralization was significantly higher at the later time points, day 28 and day 56. Reverse transcription-polymerase chain reaction confirmed the results of microarray analysis in selected genes. CONCLUSION: GC excess is associated with early activation of genes associated with osteoclastogenesis and adipogenesis and a later suppression of genes associated with osteogenesis and mineralization. Novel interventions with agents that modulate either Wnt signaling or mineralization may be effective in GC-induced osteoporosis. PMID- 18512789 TI - Effects of long-term strontium ranelate treatment on the risk of nonvertebral and vertebral fractures in postmenopausal osteoporosis: Results of a five-year, randomized, placebo-controlled trial. AB - OBJECTIVE: This study was undertaken to assess the effect of strontium ranelate on nonvertebral and vertebral fractures in postmenopausal women with osteoporosis in a 5-year, double-blind, placebo-controlled trial. METHODS: A total of 5,091 postmenopausal women with osteoporosis were randomized to receive either strontium ranelate at 2 gm/day or placebo for 5 years. The main efficacy criterion was the incidence of nonvertebral fractures. In addition, incidence of hip fractures was assessed, by post hoc analysis, in the subset of 1,128 patients who were at high risk of fractures (age 74 years or older with lumbar spine and femoral neck bone mineral density T scores -2.4 or less). The incidence of new vertebral fractures was assessed, using the semiquantitative method described by Genant, in the 3,646 patients in whom spinal radiography (a nonmandatory procedure) was performed during the course of the study. Fracture data were analyzed using the Kaplan-Meier survival method. RESULTS: Of the 5,091 patients, 2,714 (53%) completed the study up to 5 years. The risk of nonvertebral fracture was reduced by 15% in the strontium ranelate group compared with the placebo group (relative risk 0.85 [95% confidence interval 0.73-0.99]). The risk of hip fracture was decreased by 43% (relative risk 0.57 [95% confidence interval 0.33 0.97]), and the risk of vertebral fracture was decreased by 24% (relative risk 0.76 [95% CI 0.65-0.88]) in the strontium ranelate group. After 5 years, the safety profile of strontium ranelate remained unchanged compared with the 3-year findings. CONCLUSION: Our findings indicate that treatment of postmenopausal osteoporosis with strontium ranelate results in a sustained reduction in the incidence of osteoporotic nonvertebral fractures, including hip fractures, and vertebral fractures over 5 years. PMID- 18512790 TI - Aromatase expression in osteoarthritic and osteoporotic bone. AB - OBJECTIVE: Estrogen deprivation is a central mechanism in the development of osteoporosis with aging. Results from recent studies also suggest the involvement of estrogens in the pathophysiology of osteoarthritis (OA). Aromatization of androgenic precursors in peripheral tissue is the main source of estrogens in postmenopausal women and in men. However, the importance of aromatase expression in bone is a subject of controversy. This study was undertaken to determine aromatase expression in bone samples from patients with hip fracture and patients with OA. METHODS: We studied 104 patients with hip fracture (n = 60) or primary hip OA (n = 44). Aromatase expression was determined in trabecular bone samples from the femoral neck and in osteoblast cultures grown by the primary explant technique (n = 62), using real-time reverse transcriptase-polymerase chain reaction. RESULTS: Aromatase RNA was detected in bone samples at levels similar to those found in adipose tissue. Transcript levels were significantly lower in bone tissue samples obtained from patients with OA than in those obtained from patients with fracture (P = 0.00001). Likewise, primary cultures of osteoblast cells from OA patients revealed lower aromatase expression than those of cells from fracture patients (P = 0.012). Results were independent of age or sex differences. CONCLUSION: Our findings indicate that the aromatase gene is expressed in bone tissue in high amounts, similar to those found in adipose tissue, but transcript levels are lower in tissue samples and osteoblast cultures from patients with OA than in those from patients with hip fracture. Since estrogens may help to prevent local cartilage degradation, it can be speculated that such a reduced expression of aromatase could facilitate the development of OA. PMID- 18512791 TI - Age at onset-dependent presentations of premature hip osteoarthritis, avascular necrosis of the femoral head, or Legg-Calve-Perthes disease in a single family, consequent upon a p.Gly1170Ser mutation of COL2A1. AB - OBJECTIVE: To identify the genetic abnormality responsible for osteoarthritis (OA), avascular necrosis (AVN) of the femoral head, and Legg-Calve-Perthes disease in a single family, and to determine factors responsible for the distinct phenotypes manifested by different family members. METHODS: Forty-two members of a 5-generation family were recruited and investigated. Diagnosis was made by independent orthopedic surgeons and radiologists. Histopathologic changes of the diseased tissue were examined. Linkage analysis was performed with markers spanning the COL2A1 locus. Haplotypes were constructed and mutation of the gene was detected. Structures of the wild-type and mutant proteins were modeled. RESULTS: Sixteen affected members were identified (5 with isolated precocious hip OA, 6 with AVN of the femoral head, and 5 with Legg-Calve-Perthes disease). A p.Gly1170Ser mutation of COL2A1 cosegregated with the 3 diseases and was absent in controls. Of note, age at onset in relation to the closure status of the femoral head epiphysis was associated with the diseases, with Legg-Calve-Perthes disease presenting prior to closure (at ages 6-14 years), AVN of the femoral head presenting during closure (at ages 15-18 years), and precocious OA of the hip presenting after closure (at ages 21-34 years). Molecular modeling predicted that the serine-to-glycine substitution loosens the helical structure of the protein. CONCLUSION: The p.Gly1170Ser mutation of COL2A1 in the family described is responsible for pathology confined to the hip joint, which presents as isolated precocious hip OA, AVN of the femoral head, or Legg-Calve-Perthes disease. Age at onset in relation to closure of the femoral head epiphysis appears to be a critical factor in determining disease pattern. PMID- 18512793 TI - A diagnostic score for molecular analysis of hereditary autoinflammatory syndromes with periodic fever in children. AB - OBJECTIVE: To identify a set of clinical parameters that can predict the probability of carrying mutations in one of the genes associated with hereditary autoinflammatory syndromes. METHODS: A total of 228 consecutive patients with a clinical history of periodic fever were screened for mutations in the MVK, TNFRSF1A, and MEFV genes, and detailed clinical information was collected. A diagnostic score was formulated based on univariate and multivariate analyses in genetically positive and negative patients (training set). The diagnostic score was validated in an independent set of 77 patients (validation set). RESULTS: Young age at onset (odds ratio [OR] 0.94, P = 0.003), positive family history of periodic fever (OR 4.1, P = 0.039), thoracic pain (OR 4.6, P = 0.05), abdominal pain (OR 33.1, P < 0.001), diarrhea (OR 3.3, P = 0.028), and oral aphthosis (OR 0.2, P = 0.007) were found to be independently correlated with a positive genetic test result. These variables were combined in a linear score whose ability to predict a positive result on genetic testing was validated in an independent data set. In this latter set, the diagnostic score revealed high sensitivity (82%) and specificity (72%) for discriminating patients who were genetically positive from those who were negative. In patients with a high probability of having a positive result on genetic testing, a regression tree analysis provided the most reasonable order in which the genes should be screened. CONCLUSION: The proposed approach in patients with periodic fever will increase the probability of obtaining positive results on genetic testing, with good specificity and sensitivity. Our results further help to optimize the molecular analysis by suggesting the order in which the genes should be screened. PMID- 18512794 TI - Enhanced osteoclastogenesis in patients with tophaceous gout: urate crystals promote osteoclast development through interactions with stromal cells. AB - OBJECTIVE: To analyze cellular mechanisms of bone erosion in gout. METHODS: Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) from patients with gout were analyzed for the presence of osteoclast precursors. Fixed tophus and bone samples were analyzed by immunohistochemistry. Mechanisms of osteoclastogenesis were studied by culturing murine preosteoclast RAW 264.7 cells, bone marrow stromal ST2 cells, and human synovial fibroblasts with monosodium urate monohydrate (MSU) crystals. RESULTS: PBMCs from patients with severe erosive gout had the preferential ability to form osteoclast-like cells in culture with RANKL and monocyte colony-stimulating factor (M-CSF). The number of PBMC-derived tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells strongly correlated with the number of tophi (r = 0.6296, P = 0.630). Patients with severe erosive and tophaceous gout also had higher circulating concentrations of RANKL and M-CSF. Furthermore, greater numbers of TRAP-positive multinucleated cells were cultured from SFMCs derived from gouty knee effusions than from paired PBMCs (P = 0.004). Immunohistochemical analysis demonstrated numerous multinucleated cells expressing osteoclast markers within tophi and at the interface between soft tissue and bone. MSU crystals did not directly promote osteoclast formation from RAW 264.7 cells in vitro. However, MSU crystals inhibited osteoprotegerin gene and protein expression in ST2 cells and human synovial fibroblasts, without significantly altering RANKL gene expression. Conditioned medium from ST2 cells cultured with MSU crystals promoted osteoclast formation from RAW 264.7 cells in the presence of RANKL. CONCLUSION: Chronic tophaceous and erosive gout is characterized by enhanced osteoclast development. These data provide a rationale for the study of osteoclast-targeted therapies for the prevention of bone damage in chronic gout. PMID- 18512795 TI - Possible correlation between reversible and irreversible lesions in familial Mediterranean fever, soluble Fas, and C5a inhibitor activity: comment on the letter by Yasui and Yamazaki. PMID- 18512796 TI - Crystal-induced neutrophil activation: X. Proinflammatory role of the tyrosine kinase Tec. AB - OBJECTIVE: Monosodium urate monohydrate (MSU) crystals are among the most potent proinflammatory stimuli, and an innate immune inflammatory response to the crystal surface is involved in the pathogenesis of gouty arthritis. Release of the crystals into the joint cavity promotes an acute inflammation characterized by massive infiltration of neutrophils, which leads to tissue damage. The aim of the present study was to assess the involvement of the tyrosine kinase Tec in MSU crystal-initiated transduction events in human neutrophils. METHODS: Immunoprecipitation and immunoblotting techniques were used for the cellular signaling studies. Chemotaxis and enzyme-linked immunosorbent assay techniques were used for the functional studies. Silencing of Tec expression using specific small interfering RNA was also performed. RESULTS: MSU crystals induced the phosphorylation and activation of Tec in a Src-dependent manner. This activation was necessary for the MSU crystal-induced secretion of interleukin-1beta (IL 1beta) and IL-8 and for the generation of chemotactic activity in supernatants of MSU crystal-stimulated neutrophils. In addition, colchicine, an effective drug for the treatment of gout, inhibited the MSU crystal-induced tyrosine phosphorylation of Tec, thus modulating its kinase activity. CONCLUSION: Our findings show that Tec is the principal kinase of the Tec family that plays a major role in the responses of human neutrophils to MSU crystals, which are likely to be involved in the initiation and perpetuation of gout. Our results suggest that the specific inhibition of Tec during the acute phase of MSU crystal induced inflammation may be considered for the treatment of gouty arthritis. PMID- 18512797 TI - The inflammatory disease-associated variants in IL12B and IL23R are not associated with rheumatoid arthritis. PMID- 18512798 TI - Clinical Image: lipoma arborescens: a rare cause of recurrent knee effusion. PMID- 18512799 TI - Reply to letter by Lippi et al commenting on the relationship between serum vitamin D levels and levels of inflammation markers. PMID- 18512800 TI - Intensive keyboard use and carpal tunnel syndrome: comment on the article by Atroshi et al. PMID- 18512803 TI - MEFV mutations and palindromic rheumatism: comment on the article by Canete et al. PMID- 18512805 TI - An alternative, autonomic rationale for decreased risk of myocardial infarction in patients with rheumatoid arthritis responsive to anti-tumor necrosis factor therapy: comment on the article by Dixon et al. PMID- 18512807 TI - Potential involvement of TRAIL in Treg cell-mediated osteoclast suppression: comment on the article by Zaiss et al. PMID- 18512809 TI - Functionally relevant variations of the interleukin-10 gene associated with antineutrophil cytoplasmic antibody-negative Churg-Strauss syndrome, but not with Wegener's granulomatosis. AB - OBJECTIVE: Wegener's granulomatosis (WG) and Churg-Strauss syndrome (CSS) belong to the heterogeneous group of antineutrophil cytoplasmic antibody (ANCA) associated vasculitides. Current understanding of their pathogenesis and genetic background is limited. Expression levels of interleukin-10 (IL-10), a potent and pleiotropic cytokine, are largely determined by variations in the gene encoding the IL-10 precursor. This study was undertaken to determine the impact of IL10 polymorphisms on the pathogenesis of both WG and CSS in large cohorts. METHODS: Three single-nucleotide polymorphisms (SNPs) tagging the promoter haplotypes of the IL10 gene (IL10 -3575, IL10 -1082, and IL10 -592) were analyzed in 403 patients with WG and 103 patients with CSS as well as 507 matched control subjects from Germany. In addition, 3 informative SNPs in other parts of IL10 were genotyped. RESULTS: None of the markers or their haplotypes was associated with WG or any of its subgroups classified according to ANCA status, sex, or presence of further WG genetic risk factors. In contrast, the IL10 -3575/-1082/ 592 TAC haplotype, part of the extended ancient haplotype IL10.2, was highly significantly associated with ANCA-negative CSS (chi2 = 19.14, P = 0.000012, corrected P = 0.0003, odds ratio 2.16, 95% confidence interval 1.52-3.06). CONCLUSION: These findings challenge those from previous studies of IL10 in WG and provide further evidence that CSS and WG have distinct genetic backgrounds. Because the IL10.2 haplotype has been correlated reproducibly with increased IL10 expression, the possible role of IL-10 in the pathogenesis of ANCA-negative CSS needs to be further elucidated. PMID- 18512810 TI - Direct B cell stimulation by dendritic cells in a mouse model of lupus. AB - OBJECTIVE: Dendritic cells (DCs) play a major role in regulating lymphocytes, including B cells, and defective DC functions have been implicated in lupus. The purpose of this study was to assess the contribution of DCs to B cell hyperactivity in the B6.Sle1.Sle2.Sle3 (B6.TC) murine lupus model. METHODS: We compared the effects of B6 and B6.TC bone marrow-derived DCs on naive B cells cocultured in the presence of lipopolysaccharide (LPS), anti-CD40, or anti-IgM. We measured the proliferation, antibody production, and expression of activation markers and chemokine receptors for the B cells, as well as DC cytokine production. B cell proliferation was also assessed in Transwell experiments and in response to activated DC supernatants or exosomes. The role of DC-produced cytokines was evaluated with blocking antibodies and transgenic mice. RESULTS: LPS-stimulated or anti-CD40-stimulated DCs from B6.TC mice increased B cell proliferation, antibody production, and chemokine receptor expression as compared with DCs from B6 mice. Cell-to-cell contact was not necessary for the augmented effect of the lupus-prone DCs. Anti-CD40 treatment induced a higher production of interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), IL-10, and tumor necrosis factor alpha in B6.TC DCs. Blocking these individual cytokines, however, did not abrogate the effects of B6.TC DCs. Additional experiments also ruled out involvement of BAFF, IL-12, and interferon-alpha. CONCLUSION: Activated DCs from B6.TC mice directly increase B cell effector functions. This effect depends on soluble factors released by activated DCs, but none of the single major DC produced cytokines known to affect B cells are necessary. Increased sIL-6R production suggests that increased sensitivity to IL-6 may be involved. PMID- 18512811 TI - Function of CD4+,CD25+ Treg cells in MRL/lpr mice is compromised by intrinsic defects in antigen-presenting cells and effector T cells. AB - OBJECTIVE: Naturally occurring CD4+,CD25+ Treg cells are central in the maintenance of peripheral tolerance. Impaired activity and/or a lower frequency of these cells is involved in the emergence of autoimmunity. We undertook this study to analyze relative proportions and functional alterations of Treg cells in MRL/lpr mice. METHODS: The frequency of CD4+,CD25+ T cells in the peripheral blood of healthy and autoimmune mice was compared by flow cytometry. The capacity of CD4+,CD25+ T cells to inhibit the proliferation and cytokine secretion of CD4+,CD25- T cells was assessed after polyclonal activation. RESULTS: MRL/lpr mice exhibited a normal percentage of CD4+,CD25 high T cells, and forkhead box P3 messenger RNA and protein expression in Treg cells was not altered. However, MRL/lpr Treg cells displayed a reduced capacity to suppress proliferation and to inhibit interferon-gamma secretion by syngeneic effector CD4+,CD25- T cells, as compared with syngeneic cocultures of CBA/J T cells. Moreover, effector MRL/lpr CD4+,CD25- T cells were substantially less susceptible to suppression even when cultured with CBA/J or MRL/lpr Treg cells. Crossover experiments led us to conclude that in MRL/lpr mice, each partner engaged in T cell regulation displays altered functions. Molecules involved in suppressive mechanisms (CTLA-4 and CD80/CD86) are underexpressed, and antigen-presenting cells (APCs) produce raised levels of interleukin-6, which is known to abrogate suppression. CONCLUSION: Our results suggest that although the frequency and phenotype of Treg cells in MRL/lpr mice are similar to those in normal mice, Treg cells in MRL/lpr mice are not properly stimulated by APCs and are unable to suppress proinflammatory cytokine secretion from effector T cells. PMID- 18512812 TI - Activated memory B cell subsets correlate with disease activity in systemic lupus erythematosus: delineation by expression of CD27, IgD, and CD95. AB - OBJECTIVE: Analysis of peripheral B cell subsets in patients with systemic lupus erythematosus (SLE) has provided evidence of specific alterations, such as an expansion of CD27++ plasma cells/blasts and transitional B cells. However, memory B cells in lupus have not been thoroughly investigated, and only recently a CD27- memory B cell subset was identified in the peripheral blood of lupus patients. Focusing on CD27- B cells, this study aimed to identify abnormalities in peripheral B cell subsets in patients with SLE. METHODS: Three independent cohorts of lupus patients were used to characterize CD27- memory B cells, using multiparameter flow cytometry and single-cell reverse transcription-polymerase chain reaction of heavy-chain transcripts. RESULTS: We identified a homogeneous subset of CD27-,IgD-,CD95+ memory B cells with an activated phenotype that was increased in patients with disease flares and that correlated with disease activity and serologic abnormalities. In contrast, the entire subset of CD27-,IgD B cells was found to be heterogeneous, did not correlate significantly with lupus activity, and was also increased in patients with bacterial infections. CONCLUSION: We conclude that CD95 is a useful marker to identify CD27- memory B cells with an activated phenotype, which might serve as a biomarker for lupus activity and as a target of further investigations aiming to elucidate the pathogenic potential of these cells and the mechanisms involved in the generation as well as regulation of this CD27-,IgD-,CD95+ memory B cell subset. PMID- 18512813 TI - Effective therapy for nephritis in (NZB x NZW)F1 mice with triptolide and tripdiolide, the principal active components of the Chinese herbal remedy Tripterygium wilfordii Hook F. AB - OBJECTIVE: Triptolide and tripdiolide are thought to be active components of the Chinese antirheumatic herbal remedy Tripterygium wilfordii Hook F, which has been shown to be effective in treating murine lupus nephritis. This study was undertaken to examine the therapeutic effect of triptolide and tripdiolide on established lupus nephritis in (NZB x NZW)F1 mice. METHODS: (NZB x NZW)F1 mice were treated with vehicle, triptolide, or tripdiolide for 15 weeks beginning at the age of 29 weeks (after the development of lupus nephritis). Body weight, proteinuria, and anti-double-stranded DNA (anti-dsDNA) antibodies were monitored, and the kidney and spleen were assessed histologically. Culture supernatants of spleen mononuclear cells were assayed for cytokines. RESULTS: By 28 weeks, most (NZB x NZW)F1 mice had developed lupus nephritis. Vehicle-treated mice exhibited progressive proteinuria, hypoalbuminemia, elevated blood urea nitrogen (BUN) levels, and evidence of severe nephritis. In contrast, proteinuria and BUN levels were significantly reduced in mice treated with either triptolide or tripdiolide as compared with those treated with vehicle. There was no hypoalbuminemia or apparent evidence of lupus nephritis in mice treated with either of the 2 diterpenoids. At 44 weeks of age, the survival rate in mice treated with vehicle (35.7%) was markedly lower than that in mice treated with either triptolide (87.5%) or tripdiolide (88.2%). The mean level of anti-dsDNA antibody in mice treated with tripdiolide was lower than that in the vehicle-treated mice upon completion of the treatment course. Production of tumor necrosis factor, interleukin-6, and monocyte chemoattractant protein 1 by spleen cells was also decreased after diterpenoid therapy. CONCLUSION: Therapy with triptolide or tripdiolide significantly ameliorated lupus nephritis in (NZB x NZW)F1 mice, reduced cytokine and chemokine production, and prolonged survival. PMID- 18512814 TI - Systemic lupus international collaborating clinics renal activity/response exercise: comparison of agreement in rating renal response. AB - OBJECTIVE: To assess the degree to which physicians agree with each other and with ratings obtained with 3 existing responder indices, in rating the response to treatment of lupus nephritis. METHODS: Lupus nephritis patient medical records from 125 pairs of visits (6 months apart) were used to create renal response scenarios. Seven nephrologists and 22 rheumatologists rated each scenario as demonstrating complete response, partial response, same, or worsening. The plurality (most frequent) rating of renal response by the physicians was compared with the calculated score from the renal component of the British Isles Lupus Assessment Group (BILAG) index (original and updated [2004] version) and of the Responder Index for Lupus Erythematosus (RIFLE). The degree of agreement among the physicians was assessed by calculating intraclass correlation coefficients (ICCs). The degree of agreement between the plurality physician rating and ratings obtained with the established response indices was assessed using the kappa statistic. RESULTS: The ICC among all physicians was 0.64 (0.62 for nephrologists and 0.67 for rheumatologists). The chance-adjusted measure of agreement (kappa coefficient) between the plurality physician rating and the calculated score obtained using established indexes was 0.50 (95% confidence interval [95% CI] 0.38-0.61) for the RIFLE, 0.14 (95% CI 0.03-0.25) for the original BILAG, and 0.23 (95% CI 0.21-0.44) for the BILAG 2004. CONCLUSION: These findings indicate that rheumatologists as a group and nephrologists as a group have equal agreement in their rating of renal response. There was moderate agreement between plurality physician ratings and ratings obtained using the renal component of the RIFLE. Ratings of response using an index based on the original BILAG did not have good agreement with the plurality physician rating. PMID- 18512815 TI - Cardiac involvement in systemic sclerosis assessed by tissue-doppler echocardiography during routine care: A controlled study of 100 consecutive patients. AB - OBJECTIVE: To assess the prevalence of primary cardiac complications in a large population of patients with systemic sclerosis (SSc), using recently developed echocardiographic techniques. METHODS: We prospectively studied 100 consecutive patients (mean +/- SD age 54 +/- 14 years; 86 women) presenting with SSc without pulmonary arterial hypertension or clinical manifestations of heart failure. All patients underwent standard echocardiography, along with measurements of longitudinal velocities by tissue Doppler imaging (TDI) to assess left ventricular (LV) and right ventricular (RV) contractility and LV diastolic function. Results were compared with those in 26 age- and sex-matched healthy controls. RESULTS: Patients with SSc had a wider mean left atrial diameter and impaired relaxation compared with the controls. A trend was observed toward a smaller LV ejection fraction (EF) in the patients (mean +/- SD 64.9 +/- 0.6%) than in the controls (67.2 +/- 0.7%), as well as higher pulmonary artery pressure (mean +/- SD 33.3 +/- 0.6 mm Hg versus 30.8 +/- 1.0 mm Hg). LVEF was <55% in 7 patients versus none of the controls. Peak systolic mitral annular velocity as measured by TDI was <7.5 cm/second in 14 patients versus none of the controls (P = 0.040). Mitral annulus early diastolic velocity was <10 cm/second in 30 patients versus 2 of the controls (P = 0.022). Fifteen patients and none of the controls had reduced peak systolic tricuspid annular velocity (P = 0.039). The TDI results correlated with each other, but not with lung abnormalities or other disease characteristics. CONCLUSION: Depression of LV and RV systolic and LV diastolic function is common in patients with SSc and is due to primary myocardial involvement. Considering the major contributions of TDI, the addition of this simple technique to standard measurements may improve the detection of heart involvement in patients with SSc. PMID- 18512817 TI - Hypoxic synovial environment and expression of macrophage inflammatory protein 3gamma/CCL20 in juvenile idiopathic arthritis. AB - OBJECTIVE: Synovial inflammation is a major determinant of juvenile idiopathic arthritis (JIA) pathogenesis and is mediated by local chemokine secretion. Monocytic cells are an important source of chemokines. The purpose of this study was to investigate expression of CCL20, a macrophage inflammatory protein, in synovial fluid (SF) and SF-derived monocytic cells from JIA patients and its regulation by hypoxia, a common feature of the inflamed synovial environment. METHODS: Mononuclear cells and monocytic cells were isolated from paired SF and peripheral blood (PB) samples from JIA patients and were maintained in a hypoxic environment or subjected to reoxygenation. CCL20 concentrations in SF, PB, and culture supernatants were measured by enzyme-linked immunosorbent assay. CCL20 expression was assessed in both freshly purified and cultured cells by reverse transcriptase-polymerase chain reaction and immunocytochemistry. Hypoxia inducible factor 1alpha (HIF-1alpha) and HIF-2alpha were detected in the synovial tissue and cells of JIA patients by immunohistochemistry and Western blotting. RESULTS: CCL20 concentrations were significantly higher in SF compared with PB samples (P < 0.0001). SF mononuclear cells, but not matched PB mononuclear cells, constitutively expressed CCL20 messenger RNA. The SF monocytic cell fraction produced higher amounts of CCL20 than did SF lymphocytes, and CCL20 expression was associated with HIF positivity. Reoxygenation abrogated HIF and CCL20 expression, which was maintained in SF monocytic cells exposed to prolonged hypoxia. CONCLUSION: CCL20 is released into the SF of JIA patients, and SF monocytic cells are a major source of this chemokine. The hypoxic synovial microenvironment may directly contribute to the persistent inflammation associated with JIA by increasing CCL20 production by SF monocytic cells, thus representing a potential therapeutic target. PMID- 18512816 TI - A multicenter, randomized, double-blind, placebo-controlled trial of oral type I collagen treatment in patients with diffuse cutaneous systemic sclerosis: I. oral type I collagen does not improve skin in all patients, but may improve skin in late-phase disease. AB - OBJECTIVE: To investigate the safety and efficacy of oral bovine type I collagen (CI) treatment in patients who have had diffuse cutaneous systemic sclerosis (dcSSc; scleroderma) for 1-3 gm/day = 5 points, proteinuria >3 gm/day = 11 points, [corrected] urine red blood cell count > = 5/hpf = 3 points, [corrected] urine white blood cell count > or = 5/hpf = 1 point. [corrected] The chance-adjusted agreement between the renal response index derived from the activity score applied to the paired visits and the plurality physician response rating was 0.69 (95% confidence interval 0.59-0.79). CONCLUSION: Ratings derived from this index for rating of renal response showed reasonable agreement with physician ratings in a pilot study. The index will require further refinement, testing, and validation. A data-driven approach to create renal activity and renal response indices will be useful in both clinical care and research settings. PMID- 18512820 TI - Onset of psoriatic arthritis in patients treated with efalizumab for moderate to severe psoriasis. AB - OBJECTIVE: To investigate the nature of polyarthritis in patients with moderate to severe psoriasis undergoing treatment with efalizumab, a humanized anti-CD11a monoclonal antibody. METHODS: In a multicenter study, we retrospectively analyzed patients who developed arthritis during treatment with efalizumab. The relationship between joint manifestations and psoriatic disease was addressed by using different classification criteria for psoriatic arthritis (PsA). The course of arthritis and its response to treatment were also investigated. RESULTS: Sixteen patients developed de novo inflammatory rheumatic disease, with a mean delay of 15 weeks following the start of treatment, and with exclusive asymmetric peripheral monarthritis or oligoarthritis (8 patients), inflammatory spinal disease (1 patient), or both (7 patients), associated in some cases with enthesitis and dactylitis. All patients fulfilled at least 2 different sets of classification criteria for PsA. In most of them, an improvement in skin lesions was observed at the onset of PsA, as measured using the Psoriasis Area and Severity Index (mean score 24.88 before efalizumab versus 18.78 at the time of arthritis). Efalizumab treatment was stopped in 11 patients and was followed by the elimination of rheumatologic symptoms in 1 patient, while 8 patients required treatment with nonsteroidal antiinflammatory drugs with or without methotrexate, with 2 later being switched to tumor necrosis factor alpha inhibitors. Reintroduction of efalizumab (2 patients) was followed by a relapse of PsA. CONCLUSION: This study questions the role of efalizumab in the induction of PsA. It also emphasizes the discrepancy between the courses of psoriatic skin and joint manifestations under treatment. Prospective case-control studies are needed to accurately investigate the impact of efalizumab on PsA. PMID- 18512821 TI - Serum-free transient protein production system based on adenoviral vector and PER.C6 technology: high yield and preserved bioactivity. AB - Stable E1 transformed cells, like PER.C6, are able to grow at scale and to high cell densities. E1-deleted adenoviruses replicate to high titer in PER.C6 cells whereas subsequent deletion of E2A from the vector results in absence of replication in PER.C6 cells and drastically lowers the expression of adenovirus proteins in such cells. We therefore considered the use of an DeltaE1/DeltaE2 type 5 vector (Ad5) to deliver genes to PER.C6 cells growing in suspension with the aim to achieve high protein yield. To evaluate the utility of this system we constructed DeltaE1/DeltaE2 vector carrying different classes of protein, that is, the gene coding for spike protein derived from the Coronavirus causing severe acute respiratory syndrome (SARS-CoV), a gene coding for the SARS-CoV receptor or the genes coding for an antibody shown to bind and neutralize SARS-CoV (SARS-AB). The DeltaE1/DeltaE2A-vector backbones were rescued on a PER.C6 cell line engineered to constitutively over express the Ad5 E2A protein. Exposure of PER.C6 cells to low amounts (30 vp/cell) of DeltaE1/DeltaE2 vectors resulted in highly efficient (>80%) transduction of PER.C6 cells growing in suspension. The efficient cell transduction resulted in high protein yield (up to 60 picogram/cell/day) in a 4 day batch production protocol. FACS and ELISA assays demonstrated the biological activity of the transiently produced proteins. We therefore conclude that DeltaE1/DeltaE2 vectors in combination with the PER.C6 technology may provide a viable answer to the increasing demand for high quality, high yield recombinant protein. PMID- 18512822 TI - Length-dependent conductance of molecular wires and contact resistance in metal molecule-metal junctions. AB - Molecular wires are covalently bonded to gold electrodes--to form metal-molecule metal junctions--by functionalizing each end with a -SH group. The conductance of a wide variety of molecular junctions is studied theoretically by using first principles density functional theory (DFT) combined with the nonequilibrium Green's function (NEGF) formalism. Based on the chain-length-dependent conductance of the series of molecular wires, the attenuation factor beta is obtained and compared with the experimental data. The beta value is quantitatively correlated to the molecular HOMO-LUMO gap. Coupling between the metallic electrode and the molecular bridge plays an important role in electron transport. A contact resistance of 6.0+/-2.0 Kohms is obtained by extrapolating the molecular-bridge length to zero. This value is of the same magnitude as the quantum resistance. PMID- 18512823 TI - Polymorphs and polymorphic cocrystals of temozolomide. AB - Crystal polymorphism in the antitumor drug temozolomide (TMZ), cocrystals of TMZ with 4,4'-bipyridine-N,N'-dioxide (BPNO), and solid-state stability were studied. Apart from a known X-ray crystal structure of TMZ (form 1), two new crystalline modifications, forms 2 and 3, were obtained during attempted cocrystallization with carbamazepine and 3-hydroxypyridine-N-oxide. Conformers A and B of the drug molecule are stabilized by intramolecular amide N--HN(imidazole) and N- HN(tetrazine) interactions. The stable conformer A is present in forms 1 and 2, whereas both conformers crystallized in form 3. Preparation of polymorphic cocrystals I and II (TMZBPNO 1:0.5 and 2:1) were optimized by using solution crystallization and grinding methods. The metastable nature of polymorph 2 and cocrystal II is ascribed to unused hydrogen-bond donors/acceptors in the crystal structure. The intramolecularly bonded amide N-H donor in the less stable structure makes additional intermolecular bonds with the tetrazine C==O group and the imidazole N atom in stable polymorph 1 and cocrystal I, respectively. All available hydrogen-bond donors and acceptors are used to make intermolecular hydrogen bonds in the stable crystalline form. Synthon polymorphism and crystal stability are discussed in terms of hydrogen-bond reorganization. PMID- 18512824 TI - Time-resolved dynamic nuclear polarization enhanced NMR spectroscopy. PMID- 18512825 TI - Theoretical exploration of the oxidative properties of a [(tren Me1)CuO2]+ adduct relevant to copper monooxygenase enzymes: insights into competitive dehydrogenation versus hydroxylation reaction pathways. AB - Singlet and triplet H-transfer reaction paths from C-H and N-H bonds were examined by means of DFT and spin-flip TD-DFT computations on the [(tren Me1)CuO2]+ adduct. The singlet energy surfaces allow its evolution towards H2O2 and an imine species. Whereas N-H cleavage appears to be a radical process, C-H rupture results in a carbocation intermediate stabilized by an adjacent N atom and an electrostatic interaction with the [CuIOOH] metal core. Upon injection of an additional electron, the latter species straightforwardly forms a hydroxylated product. Based on these computational results, a new mechanistic description of the reactivity of copper monooxygenases is proposed. PMID- 18512826 TI - Label-free fluorescent probing of G-quadruplex formation and real-time monitoring of DNA folding by a quaternized tetraphenylethene salt with aggregation-induced emission characteristics. AB - Biosensing processes such as molecular beacons require non-trivial effort to covalently label or mark biomolecules. We report here a label-free DNA assay system with a simple dye with aggregation-induced emission (AIE) characteristics as the fluorescent bioprobe. 1,1,2,2-Tetrakis[4-(2-bromoethoxy)phenyl]ethene is nonemissive in solution but becomes highly emissive when aggregated. This AIE effect is caused by restriction of intramolecular rotation, as verified by a large increase in the emission intensity by increasing viscosity and decreasing temperature of the aqueous buffer solution of 1,1,2,2-tetrakis[4-(2 triethylammonioethoxy)phenyl]ethene tetrabromide (TTAPE). When TTAPE is bound to a guanine-rich DNA strand (G1) via electrostatic attraction, its intramolecular rotation is restricted and its emission is turned on. When a competitive cation is added to the G1 solution, TTAPE is detached and its emission is turned off. TTAPE works as a sensitive poststaining agent for poly(acrylamide) gel electrophoresis (PAGE) visualization of G1. The dye is highly affinitive to a secondary structure of G1 called the G-quadruplex. The bathochromic shift involved in the G1 folding process allows spectral discrimination of the G quadruplex from other DNA structures. The strong affinity of TTAPE dye to the G quadruplex structure is associated with a geometric fit aided by the electrostatic attraction. The distinct AIE feature of TTAPE enables real-time monitoring of folding process of G1 in the absence of any pre-attached fluorogenic labels on the DNA strand. TTAPE can be used as a K+ ion biosensor because of its specificity to K+-induced and -stabilized quadruplex structure. PMID- 18512827 TI - Doping of C60 fullerene peapods with lithium vapor: Raman spectroscopic and spectroelectrochemical studies. AB - Raman spectroscopy and in situ Raman spectroelectrochemistry have been applied to the study of the lithium vapor doping of C60@SWCNTs (peapods; SWCNT=single-walled carbon nanotube). A strong degree of doping was proven by the disappearance of the radial breathing mode (RBM) of the SWCNTs and by the attenuation of the tangential (TG) band intensity by two orders of magnitude. The lithium doping causes a downshift of the Ag(2) mode of the intratubular C60 by 27 cm(-1) and changes the resonance condition of the encapsulated fullerene. In contrast to potassium vapor doping, the strong downshift of the TG band was not observed for lithium doping. The peapods treated with lithium vapor remained partially doped even when they were exposed to humid air. This was reflected by a reduction in the intensity of the nanotube and the fullerene modes and by the change in the shape of the RBM band compared with that of the undoped sample. The Ag(2) mode of the intratubular fullerene was not resolved after contact of the lithium-doped sample with water. Lithium insertion into the interior of a peapod and its strong interaction with the intratubular fullerene is suggested to be responsible for the air-insensitive residual doping. This residual doping was confirmed by in situ spectroelectrochemical measurements. The TG band of the lithium-doped peapods did not undergo an upshift during the anodic doping, which points to the formation of a stable exohedral metallofullerene peapod. PMID- 18512828 TI - Observation of contraction and expansion in a bis(peptide)-based mechanical molecular actuator. AB - A novel, bis(peptide) based molecular actuator (1) has been synthesized. It is demonstrated to undergo contraction and expansion controlled by the addition and removal of Cu2+; this is demonstrated by the direct observation of a change in hydrodynamic properties by using sedimentation analysis and size exclusion chromatography. The molecule undergoes a large change in sedimentation coefficient, axial ratio, and size exclusion chromatography elution time when it binds copper. The demonstration of a controlled change in the mechanical properties of 1 make it a good starting point for the development of molecular devices that will harness changes in molecular shape and size to create molecular devices such as sensors or valves. PMID- 18512829 TI - N-tosyloxycarbamates as reagents in rhodium-catalyzed C-H amination reactions. AB - Metal nitrenes for use in C-H insertion reactions were obtained from N tosyloxycarbamates in the presence of an inorganic base and a rhodium(II) dimer complex catalyst. The C-H amination reaction proceeds smoothly, and the potassium tosylate that forms as a byproduct is easily removed by filtration or an aqueous workup. This new methodology allows the amination of ethereal, benzylic, tertiary, secondary, and even primary C-H bonds. The intramolecular reaction provides an interesting route to various substituted oxazolidinones, whereas the intermolecular reaction gives trichloroethoxycarbonyl-protected amines that can be isolated with moderate to excellent yields and that cleave easily to produce the corresponding free amine. The development, scope, and limitations of the reactions are discussed herein. Isotopic effects and the electronic nature of the transition state are used to discuss the mechanism of the reaction. PMID- 18512830 TI - Distinct columnar and lamellar liquid crystalline phases formed by new bolaamphiphiles with linear and branched lateral hydrocarbon chains. AB - A universal building block for the convergent synthesis of a wide variety of different T-shaped ternary amphiphiles was developed and used for the synthesis of a series of new liquid-crystalline materials composed of a rigid biphenyl core with polar glycerol groups at both ends and linear or branched alkyl chains in a lateral position. In addition, compounds with bulky achiral (2,4,6 trimethylphenoxy, adamantane-1-carboxylate, benzoate) or chiral (menthyl or cholesteryl) substituents attached to the end of the lateral alkyl chain were also investigated. In all cases the lateral chains were connected to the aromatic core by an ether linkage. The effect of the ether linking unit on mesophase stability and mesophase type is discussed with respect to conformational effects. The liquid-crystalline phases were investigated by polarizing microscopy, calorimetry, and X-ray diffraction of surface aligned samples. Upon enlarging the lateral chains a series of different polygonal cylinder phases was observed, which were replaced by lamellar phases and a non-cylinder hexagonal columnar phase by further increasing the size of these substituents. Remarkably, only pentagonal, hexagonal, and giant hexagonal cylinder phases could be observed, whereas mesophases composed of cylinders with a smaller number of sides are missing. No distinct chirality effects were observed for the menthyl- and cholesteryl-substituted compounds. However, the rodlike shape of the polycyclic cholesteryl core leads to a unique phase structure combining an organization of the alicyclic cholesteryl cores perpendicular to the layer planes and the aromatic biphenyl cores parallel to the layer planes. PMID- 18512831 TI - Phosphotungstic acid as a versatile catalyst for the synthesis of fragrance compounds by alpha-pinene oxide isomerization: solvent-induced chemoselectivity. AB - The remarkable effect of the solvent on the catalytic performance of H3PW12O40, the strongest heteropoly acid in the Keggin series, allows direction of the transformations of alpha-pinene oxide (1) to either campholenic aldehyde (2), trans-carveol (3), trans-sobrerol (4 a), or pinol (5). Each of these expensive fragrance compounds was obtained in good to excellent yields by using an appropriate solvent. Solvent polarity and basicity strongly affect the reaction pathways: nonpolar nonbasic solvents favor the formation of aldehyde 2; polar basic solvents favor the formation of alcohol 3; whereas in polar weakly basic solvents, the major products are compounds 4 a and 5. On the other hand, in 1,4 dioxane, which is a nonpolar basic solvent, both aldehyde 2 and alcohol 3 are formed in comparable amounts. The use of very low catalyst loading (0.005-1 mol %) and the possibility of catalyst recovery and recycling without neutralization are significant advantages of this simple, environmentally benign, and low-cost method. This method represents the first example of the synthesis of isomers from alpha-pinene oxide, other than campholenic aldehyde, with a selectivity that is sufficient for practical usage. PMID- 18512832 TI - Direct observation of mixed-valence and radical cation dimer states of tetrathiafulvalene in solution at room temperature: association and dissociation of molecular clip dimers under oxidative control. AB - We have observed the mixed-valence and radical cation dimer states of a glycoluril-based molecular clip with tetrathiafulvalene (TTF) sidewalls at low concentration (1 mM) at room temperature. This molecular clip has four consecutive anodic steps in its cyclic voltammogram, which suggests a sequential oxidation of these TTF sidewalls to generate species existing in several distinct charge states: neutral monomers, mixed-valence dimers, radical cation dimers, and fully oxidized tetracationic monomers. The observation of characteristic NIR spectroscopic absorption bands at approximately 1650 and 830 nm in spectroelectrochemistry experiments supports the presence of intermediary mixed valence and radical cation dimers, respectively, during the oxidation process. The stacking of four TTF radical cations in the dimer led to the appearance of a charge-transfer band at approximately 946 nm. Nanoelectrospray ionization mass spectrometry was used to verify the tricationic state and confirm the existence of other different charged dimers during the oxidation of the molecular clip. PMID- 18512833 TI - Polychromatic fingerprinting of excitation emission matrices. PMID- 18512834 TI - On-chip micro gas chromatograph enabled by a noncovalently functionalized single walled carbon nanotube sensor array. PMID- 18512835 TI - Stimuli-responsive luminescent liquid crystals: change of photoluminescent colors triggered by a shear-induced phase transition. PMID- 18512836 TI - Modulating the Lewis acidity of boron using a photoswitch. PMID- 18512837 TI - Selenium as a key element for highly ordered aromatic self-assembled monolayers. PMID- 18512838 TI - Predicting enthalpy of vaporization of ionic liquids: a simple rule for a complex property. PMID- 18512839 TI - Cage escape competes with geminate recombination during alkane hydroxylation by the diiron oxygenase AlkB. PMID- 18512840 TI - Tandem catalytic asymmetric Friedel-Crafts/Henry reaction: control of three contiguous acyclic stereocenters. PMID- 18512841 TI - On-chip fabrication of well-aligned and contact-barrier-free GaN nanobridge devices with ultrahigh photocurrent responsivity. PMID- 18512842 TI - Structural, electrical, and photoconductive properties of individual single crystalline tellurium nanotubes synthesized by a chemical route: doping effects on electrical structure. PMID- 18512843 TI - Simplified method for microlitre deuterium measurements in water and urine by gas chromatography-high-temperature conversion-isotope ratio mass spectrometry. AB - Deuterium (2H) in water and urine can be measured by off-line and, more recently, on-line techniques using isotope ratio mass spectrometry (IRMS). We describe a new simple on-line pyrolysis method for the analysis of 2H/1H in water and urine samples by continuous flow IRMS, normally used for 2H/1H measurements in organic compounds. A deactivated column connected the split injector to a high temperature conversion reactor (TC HD), and 0.5 microL of sample was injected. Accuracy and precision were determined with Vienna Standard Mean Ocean Water (VSMOW), Standard Light Antarctic Precipitation (SLAP), and Greenland Ice Sheet Precipitation (GISP). The range of linearity was measured with a calibration curve of enriched water from 0 up to 0.1 atom percent excess (APE) (i.e. -72 up to 6323 delta per mil (deltaD per thousand)) with a precision of <5 per thousand and accuracy ranging between 1 and 55 per thousand. Blinded reanalysis of urine samples by an equilibration device (Gas Bench) and by a dedicated pyrolysis system (TC/EA) was performed and results compared by the Bland-Altman test. Enrichments ranged between 600 and 2400 per thousand deltaD(VSMOW) with a precision of +/-5 per thousand. Urine enrichments described by our method were strongly correlated with values obtained by Gas Bench and TC/EA (p < 0.0001). There was a significant memory effect that was reduced by injecting the sample 15 times and discarding the first 10 injections, together with accurate furnace conditioning and appropriate cleaning of the syringe. Data indicate that the method is accurate, and that it can be used for water and urine deuterium determination when a Gas Bench or TC/EA instrument is not available and the amount of sample is limited. PMID- 18512844 TI - Mass defect profiles of biological matrices and the general applicability of mass defect filtering for metabolite detection. AB - Recent examples have demonstrated that the high-resolution liquid chromatography/mass spectrometry (LC/MS)-based mass defect filtering (MDF) technique was effective in selectively detecting drug metabolites regardless of their molecular weights or fragmentation patterns. The main objective of the current study was to evaluate the general applicability of MDF for drug metabolite detection in typical biological matrices. Mass defect profiles of commonly used biological matrices including plasma, urine, bile, and feces were obtained using an LTQ FT mass spectrometer and were compared with those of 115 commonly prescribed drugs. The mass defect profiles were presented as two dimensional Y-X plots with the determined mass defects of components on the y axis versus the corresponding m/z values on the x-axis. The mass defect profiles of the matrices appeared to be similar for each type of matrix across species, yet marked differences were apparent between matrices of a given species. The mass defect profiles of components in plasma, bile, and feces showed significant separation from most of the 115 drugs. The mass defect profiles of urine did not show such clean separation from that of the 115 drugs. The results suggest that MDF has a broad applicability for selective detection of drug metabolites in plasma, bile and feces although the selectivity for detecting urinary drug metabolites is not as good as in the other matrices. In addition, the mass defect profiles of the biological matrices allow for prediction of the effectiveness of MDF for certain applications, and for designing specific MDF windows for selective detection of drug metabolites. PMID- 18512845 TI - Simultaneous determination of water-soluble vitamins in selected food matrices by liquid chromatography/electrospray ionization tandem mass spectrometry. AB - A rapid, simple and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) with an electrospray ionization (ESI) source for the simultaneous analysis of fourteen water-soluble vitamins (B1, B2, two B3 vitamers, B5, five B6 vitamers, B8, B9, B12 and C) in various food matrices, i.e. maize flour, green and golden kiwi and tomato pulp, is presented here. Analytes were separated by ion-suppression reversed-phase liquid chromatography in less than 10 min and detected in positive ion mode. Sensitivity and specificity of this method allowed two important results to be achieved: (i) limits of detection of the analytes at ng g(-1) levels (except for vitamin C); (ii) development of a rapid sample treatment that minimizes analyte exposition to light, air and heat, eliminating any step of extract concentration. Analyte recovery depended on the type of matrix. In particular, recovery of the analytes in maize flour was > or =70%, with the exception of vitamin C, pyridoxal-5'-phosphate and vitamin B9 (ca 40%); with tomato pulp, recovery was > or =64%, except for vitamin C (41%); with kiwi, recovery was > or =73%, except for nicotinamide (ca. 30%). PMID- 18512846 TI - A comparison of cardiac (31)P MRS at 1.5 and 3 T. AB - (31)P MRS was evaluated on normal volunteers at 1.5 and 3 T, and the signal-to noise ratio (SNR) of the two field strengths was calculated. The in vivo spin lattice, T(1), relaxation times for PCr and gamma-ATP, which are essential for correcting for the effects of radiofrequency saturation on the PCr/ATP ratio, were determined at 3 T. The T(1) values for six volunteers were 3.8 +/- 0.7 s for PCr (mean +/- SD) and 2.4 +/- 1.1 s for gamma-ATP, which are similar to reported values at 1.5 T, allowing us to use protocols developed at 1.5 T at the new clinical field strength of 3 T. Direct comparison between 1.5 T and 3 T in the same 10 subjects, using coils of identical geometry and identical pulse sequences gave a mean SNR for PCr at 3 T which was 206 +/- 94% of that at 1.5 T. The linewidth for PCr increased from 13 +/- 6 Hz at 1.5 T to 22 +/- 12 Hz at 3 T. The coefficient of variation in the measurement of PCr/ATP, based on the Cramer-Rao lower bounds, was reduced from 32 +/- 25% at 1.5 T to 18 +/- 13% at 3 T. Thus, (31)P MRS at 3 T is greatly improved by the increase in SNR compared with acquisitions at 1.5 T because of the higher field strength. PMID- 18512847 TI - Comparative analysis of the gas-phase reactions of cylindrospermopsin and the difference in the alkali metal cation mobility. AB - Cylindrospermopsin (CYN) belongs to a group of toxins produced by several strains of freshwater cyanobacteria. It is a compact zwitterionic molecule composed of a uracil section and a tricyclic guanidinium portion with a primarily hepatotoxic effect. Using low multi-stage and high-resolution mass spectrometry, the gas phase reactions of this toxin have been investigated. Our data show that collision-induced dissociation (CID) spectra of CYN are dominated by neutral losses, and three major initial fragmentation pathways are clearly distinguishable. Interestingly, comparative analysis of protonated and cationizated molecules showed a significant difference in the balance of the SO3 and terminal ring elimination. These data indicate that the differential ion mobility of H+, Li+, Na+ and K+ leads to different fragmentation pathways, giving rise to mass spectra with different profiles. PMID- 18512848 TI - Isobaric metabolite interferences and the requirement for close examination of raw data in addition to stringent chromatographic separations in liquid chromatography/tandem mass spectrometric analysis of drugs in biological matrix. AB - In addition to matrix effects, common interferences observed in liquid chromatography/tandem mass spectrometry (LC/MS/MS) analyses can be caused by the response of drug-related metabolites to the multiple reaction monitoring (MRM) channel of a given drug, as a result of in-source reactions or decomposition of either phase I or II metabolites. However, it has been largely ignored that, for some drugs, metabolism can lead to the formation of isobaric or isomeric metabolites that exhibit the same MRM transitions as parent drugs. The present study describes two examples demonstrating that interference caused by isobaric or isomeric metabolites is a practical issue in analyzing biological samples by LC/MS/MS. In the first case, two sequential metabolic reactions, demethylation followed by oxidation of a primary alcohol moiety to a carboxylic acid, produced an isobaric metabolite that exhibits a MRM transition identical to the parent drug. Because the drug compound was rapidly metabolized in rats and completely disappeared in plasma samples, the isobaric metabolite appeared as a single peak in the total ion current (TIC) trace and could easily be quantified as the drug since it was eluted at a retention time very close to that of the drug in a 12 min LC run. In the second example, metabolism via the ring-opening of a substituted isoxazole moiety led to the formation of an isomeric product that showed an almost identical collision-induced dissociation (CID) MS spectrum as the original drug. Because two components were co-eluted, the isomeric product could be mistakenly quantified and reported by data processing software as the parent drug if the TIC trace was not carefully inspected. Nowadays, all LC/MS data are processed by computer software in a highly automated fashion, and some analysts may spend much less time to visually examine raw TIC traces than they used to do. Two examples described in this article remind us that quality data require both adequate chromatographic separations and close examination of raw data in LC/MS/MS analyses of drugs in biological matrix. PMID- 18512849 TI - Generation and collision-induced dissociation of ammonium tetrafluoroborate cluster ions. AB - Singly and doubly charged cluster ions of ammonium tetrafluoroborate (NH4BF4) with general formula [(NH4BF4)nNH4]+ and [(NH4BF4)m(NH4)2]2+, respectively, were generated by electrospray ionization (ESI) and their fragmentation examined using collision-induced dissociation (CID) and ion-trap tandem mass spectrometry. CID of [(NH4BF4)nNH4]+ caused the loss of one or more neutral NH4BF4 units. The n = 2 cluster, [(NH4BF4)2NH4]+, was unique in that it also exhibited a dissociation pathway in which HBF4 was eliminated to create [(NH4BF4)(NH3)NH4]+. Dissociation of [(NH4BF4)m(NH4)2]2+ occurred through two general pathways: (a) 'fission' to produce singly charged cluster ions and (b) elimination of one or more neutral NH4BF4 units to leave doubly charged product ions. CID profiles, and measurements of changing precursor and product ion signal intensity as a function of applied collision voltage, were collected for [(NH4BF4)nNH4]+ and compared with those for analogous [(NaBF4)nNa]+ and [(KBF4)nK]+ ions to determine the influence of the cation on the relative stability of cluster ions. In general, the [(NH4BF4)nNH4]+ clusters were found to be easier to dissociate than both the sodium and potassium clusters of comparable size, with [(KBF4)nK]+ ions the most difficult to dissociate. PMID- 18512850 TI - The gas-phase interactions of TEMPO radicals with Fe+ ions. AB - The gas-phase ligation of the 2,2,6,6-tetramethylpiperidyl-1-oxide (TEMPO) radical (1) and its 4-hydroxy derivative (2) with Fe+ ions in a 3 T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer was investigated. Triple ligation may occur: the first ligation produces a transient species prone to either charge exchange or a stable second ligation; the third ligand adds slowly, with fragmentation. 1 and 2 differ in that 1 binds exclusively at the nitroxyl oxygen while 2 also binds at the OH site after the loss of a H-radical. Calculations combined with steric considerations support such a mechanism for 2. The site and the mechanism of the important side reaction of 1 that involves OH addition from a water impurity to yield an FeR2 + species remain unexplained. PMID- 18512851 TI - Surface roughening of a non-tapered open tubular emitter for improved electrospray ionization mass spectrometry performance at low flow rates. AB - A non-tapered open tubular emitter with 75 microm internal diameter (i.d.) and 360 microm external diameter (o.d.) was developed by simply grinding the exit aperture of a fused-silica capillary. The roughened emitter, with a relatively large aperture, generates stable electrospray signals (generally <5% relative standard deviation (RSD) for most conditions studied) at less than 500 nL/min flow rates, and was characterized with atomic force microscopy. The surface treatment greatly extends the operational range of an open tubular emitter to lower flow rates, compared to that of a cleaved capillary with similar dimensions. The stabilized nanoelectrospray is attributed to the increased surface roughness and modified wetting characteristics of the emitter exit resulting from grinding. Electrospray performance was evaluated, and as a result of the enhanced sensitivity from a roughened emitter, five femtomoles of leucine enkephalin were detected at a 50 nL/min flow rate with a signal to noise (S/N) ratio of 48. Furthermore, trypsin-digested bovine serum albumin (BSA) was used to demonstrate the application of the emitter in protein identification, giving a sequence coverage of 60%. These emitters are robust, and may become a facile alternative to tapered emitters at moderate nano flow rates (e.g. 50 to 500 nL/min). PMID- 18512852 TI - Correlation between nuchal translucency and nuchal skin-fold measurements in Down syndrome and unaffected fetuses. AB - OBJECTIVE: To assess whether there is a correlation between nuchal translucency (NT) and nuchal skin-fold (NF) measurements, in Down syndrome and in normal pregnancies. METHODS: Nineteen Down syndrome and 224 normal fetuses underwent ultrasound sequential examinations at 11-13 weeks and 14-28 weeks' gestation. NT was measured at the earlier examination and NF at the later one. Both markers were expressed in multiples of the normal gestation-specific median (MoM). The affected cases had been referred to us for termination of pregnancy; NT had been measured locally and NF was measured at our center prior to the procedure. All unaffected pregnancies were scanned routinely at our center. RESULTS: There was no statistically significant correlation between NT and NF, in either the Down syndrome (r = 0.076, P = 0.76) or the unaffected (r = - 0.021, P = 0.76) pregnancies. The median NF value in Down syndrome fetuses was 1.538 MoM, compared with 0.990 in unaffected fetuses (P < 0.0001). Gaussian modeling with parameters from a published meta-analysis, updated to include the current series, predicted a 91% detection rate of Down syndrome for a 5% false-positive rate when NF replaced second-trimester biochemistry in a sequential contingent screening strategy. CONCLUSION: While this study cannot exclude a small correlation between NT and NF, and the Down syndrome karyotype was known at the time of the NF scan, the markers can be considered as independent determinants of Down syndrome risk. Modeling suggests that sequential contingent screening incorporating NF is an effective screening strategy, although this needs to be confirmed in a prospective study. PMID- 18512853 TI - Prenatal diagnosis of thrombosis of the dural sinuses: report of six cases, review of the literature and suggested management. AB - OBJECTIVES: To describe and assess the sonographic findings, evolution and clinical implications of thrombosis of the fetal dural sinuses. METHODS: We compiled a multicenter report of the outcomes of five cases with a prenatal diagnosis of thrombosis of the dural sinuses, and one case in which thrombosis of the dural sinus was diagnosed at necroscopy after termination of pregnancy. Prognostic factors are discussed, and suggestions made for prenatal and postnatal management. RESULTS: The mean (range) gestational age at diagnosis of thrombosis of the dural sinuses in the five cases in which it was made prenatally was 25.2 (22-31) weeks. In these five cases, diagnosis was made by sonography and confirmed by magnetic resonance imaging (MRI), which showed a blood clot in the region of the torcular herophili. Three of the six cases delivered vaginally with favorable sonographic findings, and normal clinical neurological development. Two pregnancies were terminated at the request of the parents. In one of these cases the prognosis was poor, with signs of fetal decompensation or cardiac failure; the pregnancy was terminated and necropsy revealed thrombosis of the occipital dural sinuses associated with a hemangioma. One infant, in whom the thrombosis developed in conjunction with a dural sinus malformation, died at 4 months of age. CONCLUSIONS: Thrombosis of the cerebral venous circulation can occur antenatally and is detectable by fetal real-time and color Doppler ultrasound examination. A review of the literature supports targeted evaluation of the fetus by serial ultrasound imaging and MRI to help guide the diagnosis, and to improve the counseling and management of such cases. Partial or total regression, isolated abnormality, absence of fetal decompensation or signs of cardiac failure and favorable clinical evolution are suggestive of favorable prognosis. In such cases, non-interventional neonatal management is recommended. PMID- 18512854 TI - Frontomaxillary facial angle in screening for trisomy 21 at 11 + 0 to 13 + 6 weeks. AB - OBJECTIVE: Trisomy 21 is associated with a flat face, which can now be quantified by measurement of the frontomaxillary facial (FMF) angle. The aim of this study was to examine whether in trisomy 21 fetuses fetal nuchal translucency (NT) thickness and maternal serum free ss-human chorionic gonadotropin (ss-hCG) and pregnancy-associated plasma protein-A (PAPP-A) are independent of the FMF angle, and to estimate the performance of a first-trimester screening test for trisomy 21 that includes measurement of the FMF angle. METHODS: This was a prospective study in singleton pregnancies at 11 + 0 to 13 + 6 weeks of gestation in which three-dimensional volumes of the fetal head were obtained and measurement of the FMF angle performed immediately before fetal karyotyping by chorionic villus sampling (CVS). The women chose to have CVS after risk assessment by a combination of maternal age, fetal NT thickness and maternal serum free ss-hCG and PAPP-A. Regression analysis was used to examine the significance of the association within the euploid and within the trisomy 21 fetuses between the deviation from the normal median in FMF angle and the deviation in NT, free ss hCG and PAPP-A. We estimated the detection rate (DR) and false positive rate (FPR) of first-trimester screening for trisomy 21 by measuring the FMF angle in all cases and of an alternative policy in which first-stage screening is by fetal NT and maternal serum biochemistry in all patients, followed by second-stage assessment of FMF angle only in those with an intermediate risk (1 in 51 to 1 in 1000) after the first stage. RESULTS: The FMF angle was measured in 782 euploid and 108 trisomy 21 fetuses. In the euploid fetuses the mean FMF angle decreased linearly with CRL from 83.5 degrees at a crown-rump length (CRL) of 45 mm to 76.4 degrees at a CRL of 84 mm. In the euploid fetuses the mean delta FMF angle was 0.0 (SD, 4.264) degrees and the respective values in the trisomy 21 fetuses were 7.172 (SD, 4.092) degrees . Incorporating the FMF angle in first-trimester combined screening increased the estimated DR from 90 to 94% at an FPR of 5% and from 85 to 92% at an FPR of 3%. In two-stage screening it would be necessary to measure the FMF angle in 12% of cases and the DRs would be 93 and 91% at FPRs of 5 and 3%, respectively. CONCLUSIONS: Measurement of the FMF angle improves the performance of first-trimester screening for trisomy 21. PMID- 18512855 TI - Indirect resolution of baclofen enantiomers from pharmaceutical dosage form by reversed-phase liquid chromatography after derivatization with Marfey's reagent and its structural variants. AB - A high-performance liquid chromatographic (HPLC) method was developed for chiral assay of baclofen enantiomers in pharmaceutical formulations using an indirect approach. Baclofen enantiomers were derivatized with Marfey's reagent (FDNP-L-Ala NH2) and its structural variants FDNP-L-Phe-NH2, FDNP-L-Val-NH2, FDNP-L-Leu-NH2 and FDNP-L-Pro-NH2. The resultant diastereomers were separated on RP-TLC [triethylammonium phosphate buffer (pH 4.0, 50 mm)-acetonitrile, 50:50] and on a C18 column using a linear gradient (45 min) of acetonitrile and 0.01% aqueous trifluoroacetic acid (TFA) with UV detection at 340 nm. The differences in the retention times (Delta t R) of diastereomers due to the five chiral reagents were compared. The maximum and minimum difference in retention times between separated diastereomers was for FDNP-L-Leu-NH2 and FDNP-L-Pro-NH2, respectively. The effect of flow rate, acetonitrile content and TFA concentration on resolution was studied. The method was validated for linearity, repeatability, limit of detection and limit of quantification. PMID- 18512856 TI - Application of high-speed counter-current chromatography for the isolation of antiviral eremophilenolides from Ligularia atroviolacea. AB - Ligularia is mainly distributed in the western regions of China. Most of the species have been traditionally used in folk medicine for the treatment of hepatitis B, asthma, hemoptysis and pulmonary tuberculosis. In our continuation of research on antiviral components from traditional Chinese medicine, Ligularia atroviolacea was tested for inhibitory effects on hepatitis B virus (HBV). A bioassay-guided phytochemical examination of L. atroviolacea disclosed that its ethyl acetate extract, which was made up of two eremophilenolides, showed suppressive activity on the expression of HBV surface antigen (HBsAg) in the HepG2.2.15 cell line. Then a simple and effective preparative high-speed counter current chromatography method was successfully developed for the isolation and purification of two main active metabolites, 8beta-hydroxyeremophil-3,7(11)-dien 12,8alpha;15,6alpha-diolide and 8beta-methoxyeremophil-3,7(11)-dien 12,8alpha;15,6alpha-diolide from the ethyl acetate extract of L. atroviolacea by a one-step separation using a two-phase solvent system composed of light petroleum (60-90 degrees C)-ethyl acetate-methanol-water (9:1:8:2, v/v/v). The chemical structures of the two eremophilenolides were identified by ESI-MS, (1)H NMR and (13)C-NMR analysis. The anti-HBV activity of the two purified compounds was measured; both of them showed suppressive activity on the expression of HBsAg in the HepG2.2.15 cell line. The results support the continued and expanded exploitation and utilization of L. atroviolacea. PMID- 18512857 TI - Ecstasy-related deficits in the updating component of executive processes. AB - AIMS: Research shows that users of ecstasy (MDMA) exhibit deficits in executive processes. The updating component appears to be particularly susceptible. Less is known about the precise nature of such deficits. The present study sought to determine if ecstasy-related deficits in memory updating are related to serial position of items presented, or length of the list of items. METHOD: Seventy three ecstasy/polydrug users and seventy-three non-ecstasy users completed tasks of verbal and spatial memory running memory, recalling the most recent items, in lists of varying and unknown length. Participants were categorised according to letter and spatial span (four, five or six), producing six sub-samples for analysis. RESULTS: Ecstasy-polydrug users were impaired in four out of the six sub-sample analyses. Three of these were due to impaired recall of earlier serial positions. CONCLUSIONS: The results of the present study provide further support for updating deficits in ecstasy-polydrug users. The results are suggestive of a breakdown in the maintenance of information in working memory in terms of chunking; it appears that ecstasy/polydrug users are as able as non-ecstasy users to form memory 'chunks' from the items, but that such chunks are not retained as effectively. PMID- 18512858 TI - Direct catalytic asymmetric three-component Kabachnik-Fields reaction. PMID- 18512859 TI - Generation of novel pikromycin antibiotic products through mutasynthesis. AB - The pikromycin polyketide synthase (PKS) of S. venezuelae, which consists of one loading module and six extension modules, is responsible for the formation of the hexaketide narbonolide, a key intermediate in the biosynthesis of the antibiotic pikromycin. S. venezuelae strains in which PikAI, which houses the loading domain and first two modules of the PKS, is either absent or catalytically inactive, produce no pikromycin product. When these strains are grown in the presence of a synthetically prepared triketide product, activated as the N-acetylcysteamine thioester, pikromycin yields are restored to as much as 11 % of that seen in the wild-type strain. Feeding analogues of the triketide intermediate provides pikromycin analogues bearing different alkyl substituents at C13 and C14. One of these analogues, Delta(15,16)-dehydropikromycin, exhibits improved antimicrobial activity relative to pikromycin. PMID- 18512860 TI - Microbial synthesis of CdS nanocrystals in genetically engineered E. coli. PMID- 18512861 TI - Enrichment of cell-targeting and population-specific aptamers by fluorescence activated cell sorting. PMID- 18512862 TI - On the mechano-chiral effect of vortical flows on the dichroic spectra of 5 phenyl-10,15,20-tris(4-sulfonatophenyl)porphyrin J-aggregates. AB - Phase-modulated ellipsometry of the J-aggregates of the title porphyrin shows that the material gives a true CD signal. This confirms that there is a real chiral transfer by mechanical forces, mediated by shear gradient flows, from the macroscopic to the electronic transition level. Dislocations in the structure of the aggregate could justify the formation of chirality at the level of the electronic transitions once the mesophases can be sculptured by hydrodynamic gradient flows. PMID- 18512863 TI - A general organocatalytic enantioselective malonate addition to alpha,beta unsaturated enones. AB - A general enantioselective organocatalytic conjugate addition procedure of a variety of malonates to alpha,beta-unsaturated enone systems is presented. The reaction is efficiently catalysed by the pyrrolidinyl tetrazole catalyst 1. Cyclic, acyclic and aromatic enones can be used and the reaction with ethyl malonates 3 b provides the Michael addition products in high yields with good to excellent enantioselectivities. Since only 1.5 equivalents of malonate are used as a reagent, the reaction is readily scaled and practical to operate. Furthermore, the malonate addition products can be easily mono decarboxylated without loss in enantiomeric excess by enzymatic or sodium hydroxide mediated methods. PMID- 18512864 TI - Age related mitochondrial degenerative disorders in humans. AB - Disorders caused by mitochondrial respiratory chain deficiency due to mutations in mitochondrial DNA have varied phenotypes but many involve neurological features often associated with cell loss within specific brain regions. These disorders, along with the increasing evidence of decline in mitochondrial function with ageing, have raised speculation that primary changes in mitochondria could have an important role in age-related neurodegenerative diseases such as Parkinson's disease (PD) and Alzheimer's disease (AD). Evidence supporting a role for mitochondria in common neurodegenerative diseases comes from studies with the toxin MPP+ and familial PD, which has been shown to involve proteins such as DJ-1 and Pink1 (both of which are predicted to have a role in mitochondrial function and oxidative stress). Mutations within the mitochondrial genome have been shown to accumulate with age and in common neurodegenerative diseases. Mitochondrial DNA haplogroups have also been shown to be associated with certain neurodegenerative conditions. This review covers the primary mitochondrial diseases but also discuss the potential role of mitochondria and mitochondrial DNA mutations in mitochondrial and neurodegenerative diseases, in particular in PD and in AD. PMID- 18512865 TI - Full structure of the carbohydrate chain of the lipopolysaccharide of Providencia rustigianii O34. AB - A lipopolysaccharide isolated from an opportunistic pathogen of the Enterobacteriaceae family Providencia rustigianii O34 was found to be a mixture of R-, SR-, and S-forms consisting of a lipid moiety (lipid A) that bears a core oligosaccharide, a core with one O-polysaccharide repeating unit attached, and a long-chain O-polysaccharide, respectively. The corresponding carbohydrate moieties were released from the lipopolysaccharide by mild acid hydrolysis and studied by sugar and methylation analyses along with one- and two-dimensional NMR spectroscopy and high-resolution electrospray ionization mass spectrometry. As a result, the structures of the core and the O-polysaccharide were established, including the structure of the biological repeating unit (an oligosaccharide that is preassembled and polymerized in biosynthesis of the O-polysaccharide), as well as the mode of the linkage between the O-polysaccharide and the core. Combining the structure of the carbohydrate moiety thus determined and the known structure of lipid A enabled determination of the full lipopolysaccharide structure of P. rustigianii O34. PMID- 18512866 TI - Solid-phase organic tagging resins for labeling biomolecules by 1,3-dipolar cycloaddition: application to the synthesis of a fluorescent non-peptidic vasopressin receptor ligand. AB - Two novel solid-phase organic tagging (SPOrT) resins were synthesized to facilitate the labeling of peptides and small organic compounds with a fluorescent probe. Both resins were obtained from the commercially available backbone amide linker (BAL) resin. Following the solid-phase synthesis of model compounds, a tripeptide and benzazepine, the fluorescent probe derived from Lissamine Rhodamine B was incorporated through CuI-catalyzed 1,3-dipolar cycloaddition. Final cleavage in acidic media enabled access to both types of molecules in good yield with high purity. The SPOrT resin was successfully applied to the preparation of the first non-peptidic fluorescent compound with a nanomolar affinity for the human vasopressin V2 receptor (V2R) subtype. This molecule will find application in binding assays that use polarization or fluorescence resonance energy-transfer (FRET) techniques. The SPOrT resins are also well suited for other tags and the parallel synthesis of a fluorescently tagged library for protein screening. PMID- 18512867 TI - Factors controlling the alkyne prins cyclization: the stability of dihydropyranyl cations. AB - The relative stability of the intermediates involved in the alkyne Prins cyclization and the competitive 2-oxonia-[3,3]-sigmatropic rearrangement was studied. This rearrangement was shown to occur slowly under typical alkyne Prins cyclization conditions when the allenyl oxocarbenium ion that results from the rearrangement is similar to or higher in energy than the starting alkynyl oxocarbenium ion. The formal 2-oxonia-[3,3]-sigmatropic rearrangement may be disfavored by destabilizing the resultant allenyl oxocarbenium ion or by stabilizing an intermediate dihydropyranyl cation. The trimethylsilyl group as a substituent at the alkyne and electron-withdrawing groups (CH2Cl and CH2CN) located at the alpha-position to the carbinol center are shown to be effective. DFT calculations suggest that these substituents stabilize the dihydropyranyl cations, thus leading to a more uniform reorganization of the electronic density in the ring, and do not have a direct effect on the formally positively charged carbon atom. PMID- 18512868 TI - De-racemization of enantiomers versus de-epimerization of diastereomers- classification of dynamic kinetic asymmetric transformations (DYKAT). AB - The isolation of single stereoisomers from a racemic (or diastereomeric) mixture by enzymatic or chemical resolution techniques goes in hand with the disposal of 50% (racemate) or more (diastereomeric mixtures) of the "undesired" substrate isomer(s). In order to circumvent this drawback, dynamic systems have been developed for the de-racemization of enantiomers and the de-epimerizations of diastereomers. Key strategies within this area are discussed and are classified according to their underlying kinetics, that is, dynamic kinetic resolution (DKR), dynamic kinetic asymmetric transformations (DYKAT), and hybrids between both of them. Finally, two novel types of DYKAT are defined. PMID- 18512869 TI - Structural implications of mitochondrial dynamics. AB - Mitochondrial components are continuously distributed throughout the whole chondriome of a cell by fusion and fission. Thus, a single mitochondrion represents a transient fraction of the chondriome. Mitochondrial dynamics are responsible for intracellular distribution and reaction of mitochondria to functional requirements. Dynamics occur on different levels: overall morphology, inner membrane-matrix compartment, turnover and rearrangements of mitochondrial proteins and DNA. Electron micrographs of serial sections of human umbilical vein endothelial cells reveal perinuclear mitochondria of extreme length and with branches in those cells that also have short peripheral mitochondria. Interactions of mitochondria with cytoskeletal elements are revealed in cells treated with cytochalasin D to destroy actin fibrillar structures or after disassembling microtubule by nocodazole. In the latter case mitochondria not only become immobilized, they also acquire a multiple ring structure. In F-actin disturbed cells, motility (shape changes in particular) is increased and the mitochondria become elongated. Mechanisms of how F-actin might render mitochondria immobile may involve dynamin-related protein 1 (DRP1) or interaction with anion channels. This may be responsible for the lack of mitochondrial motility in senescent cells. Fusion between mitochondria revealed local fluctuations of mitochondrial red fluorescent protein (mtRFP), indicating novel fast inner membrane reorganizations. Mitochondrial dynamics result from a complex interplay between the molecular organization of the inner membrane-matrix complex and cytoskeletal elements outside. PMID- 18512870 TI - Lipases at interfaces: unique interfacial properties as globular proteins. AB - The adsorption behavior of two globular proteins, lipase from Rhizomucor miehei and beta-lactoglobulin, at inert oil/water and air/water interfaces was studied by the pendant drop technique. The kinetics and adsorption isotherms were interpreted for both proteins in different environments. It was found that the adopted mathematical models well describe the adsorption behavior of the proteins at the studied interfaces. One of the main findings is that unique interfacial properties were observed for lipase as compared to the reference beta lactoglobulin. A folded drop with a "skinlike" film was formed for the two proteins after aging followed by compression. This behavior is normally associated with protein unfolding and covalent cross-linking at the interface. Despite this, the lipase activity was not suppressed. By highlighting the unique interfacial properties of lipases, we believe that the presented work contributes to a better understanding of lipase interfacial activation and the mechanisms regulating lipolysis. The results indicate that the understanding of the physical properties of lipases can lead to novel approaches to regulate their activity. PMID- 18512871 TI - Effect of polymer architecture on the adsorption properties of a nonionic polymer. AB - The adsorption of a linear- and bottle-brush poly(ethylene oxide (PEO))-based polymer, having comparable molecular weights, was studied by means of quartz crystal microbalance with dissipation monitoring ability (QCM-D) and AFM colloidal probe force measurements. The energy dissipation change monitored by QCM-D and the range of the steric forces obtained from force measurements demonstrated that linear PEO forms a more extended adsorption layer than the bottle-brush polymer, despite that the adsorbed mass is higher for the latter. Competitive adsorption studies revealed that linear PEO is readily displaced from the interface by the bottle-brush polymer. This was attributed to the higher surface affinity of the latter, which is governed by the number of contact points between the polymers and the interface, and the smaller loss of conformational entropy. PMID- 18512872 TI - Supercritical fluid swelling of liquid crystal films. AB - The influence of liquid and supercritical carbon dioxide and liquid propane on the structural properties of both ionic and nonionic surfactant-based liquid crystal films is discussed in this paper. Swelling of the films, measured using in situ small-angle neutron scattering (SANS), was found to be dependent on the solubility of the propane/carbon dioxide in the micelles of the respective liquid crystals. Additionally, under certain pressure conditions the structural properties of some of the films were observed to change, ultimately leading to a loss of order in the micellar arrays of the liquid crystals. PMID- 18512873 TI - Chemical and morphological consequences of acidification of pure, phosphated, and phosphonated CaO: influence of CO2 adsorption. AB - In situ Fourier transform infrared (FTIR) spectroscopy was employed to characterize the adsorption behavior (as a function of pressure or time) and surface species of CO2 molecules on pure, phosphated, and phosphonated CaO. Carbonate and bicarbonate species were found to form on the pure oxide, whereas on the phosphated and phosphonated oxide samples the carbonate species were found to substitute favorably some of the OH(-) and PO4(3-) groups thereon exposed, respectively. Before and after carbonation, the test samples were further examined by in situ FTIR spectroscopy of adsorbed pyridine species, scanning electron microscopy, and energy dispersive X-ray spectroscopy. Then they were in situ acidified by exposure to a wet atmosphere of HCl vapor at 673 K for 10 min and re-examined similarly to reveal the influence of CO2 adsorption on the chemical and morphological consequences of acidification. The results obtained show the carbonate substitution of PO4(3-) groups to enhance agglomeration of the otherwise fine, longitudinal material particles into much bulkier ones and to render the otherwise more stable phosphonate groups less stable to acid treatment than the phosphate groups. Moreover, the bulky particle agglomerates of the carbonated test samples were detectably eroded following the acid treatment. PMID- 18512874 TI - Microfluidic chip-based fabrication of PLGA microfiber scaffolds for tissue engineering. AB - In this paper, we have developed a method to produce poly(lactic- co-glycolic acid) (PLGA) microfibers within a microfluidic chip for the generation of 3D tissue engineering scaffolds. The synthesis of PLGA fibers was achieved by using a polydimethylsiloxane (PDMS)-based microfluidic spinning device in which linear streams of PLGA dissolved in dimethyl sulfoxide (DMSO) were precipitated in a glycerol-containing water solution. By changing the flow rate of PLGA solution from 1 to 50 microL/min with a sheath flow rate of 250 or 1000 microL/min, fibers were formed with diameters that ranged from 20 to 230 microm. The PLGA fibers were comprised of a dense outer surface and a highly porous interior. To evaluate the applicability of PLGA microfibers generated in this process as a cell culture scaffold, L929 fibroblasts were seeded on the PLGA fibers either as-fabricated or coated with fibronectin. L929 fibroblasts showed no significant difference in proliferation on both PLGA microfibers after 5 days of culture. As a test for application as nerve guide, neural progenitor cells were cultured and the neural axons elongated along the PLGA microfibers. Thus our experiments suggest that microfluidic chip-based PLGA microfiber fabrication may be useful for 3D cell culture tissue engineering applications. PMID- 18512875 TI - Exercising spatiotemporal control of cell attachment with optically transparent microelectrodes. AB - This paper describes a novel approach of controlling cell-surface interactions through an electrochemical "switching" of biointerfacial properties of optically transparent microelectrodes. The indium tin oxide (ITO) microelectrodes, fabricated on glass substrates, were modified with poly(ethylene glycol) (PEG) silane to make glass and ITO regions resistant to protein and cell adhesion. Cyclic voltammetry, with potassium ferricyanide serving as a redox reporter molecule, was used to monitor electron transfer across the electrolyte-ITO interface. PEG silane modification of ITO correlated with diminished electron transfer, judged by the disappearance of ferricyanide redox activity. Importantly, application of reductive potential (-1.4 V vs Ag/AgCl reference) corresponded with reappearance of typical ferricyanide redox peaks, thus pointing to desorption of an insulating PEG silane layer. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) characterization of the silanized ITO surfaces after electrical stimulation indicated complete removal of the silane layer. Significantly, electrical stimulation allowed to "switch" chosen electrodes from nonfouling to protein-adhesive while leaving other ITO and glass regions protected by a nonfouling PEG silane layer. The spatial and temporal control of biointerfacial properties afforded by our approach was utilized to micropattern proteins and cells and to construct micropatterned co-cultures. In the future, control of the biointerfacial properties afforded by this novel approach may allow the organization of multiple cell types into precise geometric configurations in order to create better in vitro mimics of cellular complexity of the native tissues. PMID- 18512876 TI - Ionic liquid induced changes in the properties of aqueous zwitterionic surfactant solution. AB - Modifying properties of aqueous surfactant solutions by addition of external additives is an important area of research. Unusual properties of ionic liquids (ILs) make them ideal candidates for this purpose. Changes in important physicochemical properties of aqueous zwitterionic N-dodecyl- N, N-dimethyl-3 ammonio-1-propanesulfonate (SB-12) surfactant solution upon addition of hydrophilic IL 1-butyl-3-methylimidazolium tetrafluoroborate, [bmim][BF 4], are reported. Dynamic light scattering results indicate a dramatic reduction in the average micellar size in the presence of [bmim][BF 4]; micellar (or micelle-like) aggregation in the presence of as high as 30 wt % [bmim][BF 4] is confirmed. Responses from fluorescence probes are used to obtain critical micelle concentration (cmc), aggregation number ( N agg), and dipolarity and microfluidity of the micellar pseudophase of aqueous SB-12 in the presence of [bmim][BF 4]. In general, increasing the amount of [bmim][BF 4] to 30 wt % results in decrease in N agg and increase in cmc. Increase in the dipolarity and the microfluidity of the probe cybotactic region within the micellar pseudophase is observed on increasing [bmim][BF 4] concentration in the solution. It is attributed to increased water penetration into the micellar pseudophase as [bmim][BF 4] is added to aqueous SB-12. It is proposed that IL [bmim][BF 4] behaves similar to an electrolyte and/or a cosurfactant when present at low concentrations and as a polar cosolvent when present at high concentrations. Electrostatic attraction between cation of IL and anion of zwitterion, and anion of IL and cation of zwitterion at low concentrations of [bmim][BF 4] is evoked to explain the observed changes. Presence of IL as cosolvent appears to reduce the efficiency of micellization process by reducing the hydrophobic effect. PMID- 18512877 TI - Coupled electrostatic, hydrodynamic, and mechanical properties of bacterial interfaces in aqueous media. AB - The interactions of bacteria with their environment are governed by a complex interplay between biological and physicochemical phenomena. The main challenge is the joint determination of the intertwined interfacial characteristics of bacteria such as mechanical and hydrodynamic softness, interfacial heterogeneity, and electrostatic properties. In this study, we have combined electrokinetics and force spectroscopy to unravel this intricate coupling for two types of Shewanella bacterial strains that vary according to the nature of their outer, permeable, charged gel-like layers. The theoretical interpretation of the bacterial electrokinetic response allows for the estimation of the hydrodynamic permeability, degree of interfacial heterogeneity, and volume charge density for the soft layer that constitutes the outer permeable part of the bacteria. Additionally, the electrostatic interaction forces between an AFM probe and the bacteria were calculated on the basis of their interfacial properties obtained from advanced soft particle electrokinetic analysis. For both bacterial strains, excellent agreement between experimental and theoretical force curves is obtained, which highlights the necessity to account for the interfacial heterogeneity of the bioparticle to interpret AFM and electrokinetic data consistently. From the force profiles, we also derived the relevant mechanical parameters in relation to the turgor pressure within the cell and the nature of the bacterial outer surface layer. These results corroborate the heterogeneous representation of the bacterial interface and show that the decrease in the turgor pressure of the cell with increasing ionic strength is more pronounced for bacteria with a thin surface gel-like layer. PMID- 18512878 TI - Stability of self-assembled monolayers on titanium and gold. AB - Methyl- and hydroxyl-terminated phosphonic acid self-assembled monolayers (SAMs) were coated on Ti from aqueous solution. Dodecyl phosphate and dodecyltrichlorosilane SAMs were also coated on Ti using solution-phase deposition. The stability of SAMs on Ti was investigated in Tris-buffered saline (TBS) at 37 degrees C using X-ray photoelectron spectroscopy, contact angle goniometry, and atomic force microscopy. For comparison purposes, a hydroxyl terminated thiol SAM was coated on Au, and its stability was also investigated under similar conditions. In TBS, a significant proportion of phosphonic acid or phosphate molecules were desorbed from the Ti surface within 1 day, while the trichlorosilane SAM on Ti or thiol SAM on Au was stable for up to 7 days under similar conditions. The stability of hydroxyl-terminated phosphonic acid SAM coated Ti and thiol SAM coated Au was investigated in ambient air and ultraviolet (UV) light. In ambient air, the phosphonic acid SAM on Ti was stable for up to 14 days, while the thiol SAM on Au was not stable for 1 day. Under UV-radiation exposure, the alkyl chains of the phosphonic acid SAM were decomposed, leaving only the phosphonate groups on the Ti surface after 12 h. Under similar conditions, decomposition of alkyl chains of the thiol SAM was observed on the Au surface accompanied by oxidation of thiolates. PMID- 18512879 TI - An affinity-based strategy for the design of selective displacers for the chromatographic separation of proteins. AB - We describe an affinity-based strategy for designing selective protein displacers for the chromatographic purification of proteins. To design a displacer that is selective for a target protein, we attached a component with affinity for the target protein to a resin-binding component; we then tested the ability of such displacers to selectively retain the target protein on a resin relative to another protein having a similar retention time. In particular, we synthesized displacers based on biotin, which selectively retained avidin as compared to aprotinin on SP Sepharose high performance resin. In addition, we have extended this approach to develop an affinity-peptide-based displacer that discriminates between lysozyme and cytochrome c. Here, a selective displacer was designed from a lysozyme-binding peptide that had been identified and optimized previously using phage-display technology. Our results suggest a general strategy for designing highly selective affinity-based displacers by identifying molecules (e.g., peptides) that bind to a protein of interest and using an appropriate linker to attach these molecules to a moiety that binds to the stationary phase. PMID- 18512880 TI - Influence of fluid flow on the deposition of soluble surfactants through receding contact lines of volatile solvents. AB - Soluble surfactants are often deposited from volatile solvents through moving contact lines. In this study, we demonstrate that altering the flow field near such a contact line fundamentally changes the deposited surfactant structure. At slow contact line speeds, the substrate emerges dry. A densely packed, tilted monolayer of surfactant is deposited along the solid-vapor interface from the rolling fluid motion at the contact line. At faster speeds, the substrate emerges with an evaporating thin film entrained on its surface. Surfactant is confined in the film in a constantly increasing concentration environment. Monodisperse crystalline islands nucleate and grow on the surface with sizes and shapes controlled by varying the deposition conditions. These results contrast with disordered deposits that result from evaporation at a pinned contact line. Our results suggest that dip-coating with control of dipping speed and evaporation rate may provide better control of deposition through contact lines of evaporating solvents. PMID- 18512881 TI - Antibacterial effects of carbon nanotubes: size does matter! AB - We provide the first evidence that the size (diameter) of carbon nanotubes (CNTs) is a key factor governing their antibacterial effects and that the likely main CNT-cytotoxicity mechanism is cell membrane damage by direct contact with CNTs. Experiments with well-characterized single-walled carbon nanotubes (SWNTs) and multiwalled carbon nanotubes (MWNTs) demonstrate that SWNTs are much more toxic to bacteria than MWNTs. Gene expression data show that in the presence of both MWNTs and SWNTs, Escherichia coli expresses high levels of stress-related gene products, with the quantity and magnitude of expression being much higher in the presence of SWNTs. PMID- 18512882 TI - Characterization of globular protein solutions by dynamic light scattering, electrophoretic mobility, and viscosity measurements. AB - In this work, physicochemical properties of two globular proteinsbovine serum albumin (BSA) having a molecular weight of 67 kDa and human serum albumin (HSA) having a molecular weight of 69 kDawere characterized. The bulk characteristics of these proteins involved the diffusion coefficient (hydrodynamic radius), electrophoretic mobility, and dynamic viscosity as a function of protein solution concentration for various pH values. The hydrodynamic radius data suggested an association of protein molecules, most probably forming compact dimers. Using the hydrodynamic diameter and the electropheretic mobility data allowed the determination of the number of uncompensated (electrokinetic) charges on protein surfaces. The electrophoretic mobility data were converted to zeta potential values, which allowed one to determine the isoelectric point (iep) of these proteins. It was found to be at pH 5.1 for both proteins, in accordance with previous experimental data and theoretical estimations derived from amino acid composition and p K values. To determine further the stability of protein solutions, dynamic viscosity measurements were carried out as a function of their bulk volume concentration for various pH values. The intrinsic viscosity derived from these measurements was interpreted in terms of the Brenner model, which is applicable to hard spheroidal particles. It was found that the experimental values of the intrinsic viscosity of these proteins were in good agreement with this model when assuming protein dimensions of 9.5 x 5 x 5 nm3 (prolate spheroid). The possibility of forming linear aggregates of association degree higher than 2 was excluded by these measurements. It was concluded that the combination of dynamic viscosity and dynamic light scattering can be exploited as a convenient tool for detecting not only the onset of protein aggregation in suspensions but also the form and composition of these aggregates. PMID- 18512883 TI - Physicochemical and biological characterization of ceramide-containing liposomes: paving the way to ceramide therapeutic application. AB - Ceramides mediate antiproliferative responses, and it has been proposed that increasing the level of ceramides in cancer cells may have a therapeutic antitumor effect. However, ceramides, because of their high "packing parameter" (PP), do not form lipid assemblies that can be dispersed in a form suitable for intravenous administration. We found that nanoliposomes containing short- or medium-chain ceramides are unstable because of their very high (>1.3) PP. To overcome this major obstacle, we included the lipopolymer 2kPEG-DSPE, which reduces the additive PP. The presence of PEG-DSPE allows the formation of highly stable (>1 year) ceramide (Cer)-containing nanoliposomes suitable for systemic administration. Using tumor cell lines, we found that the ceramide cytotoxicity was not impaired by their inclusion in nanoliposomes. The use of 14C-labeled ceramides shows that the C6Cer, but not C16Cer, was transferred from the nanoliposomes to the cells and metabolized efficiently. The difference between the two ceramides is related to the large difference between their critical aggregation concentration and was correlated with the much higher cytotoxity of liposomal C6Cer. The activity of 2kPEG-DSPE as a steric stabilizer (as previously shown for Doxil) was also confirmed for C6Cer-containing nanoliposomes. The 2kPEG DSPE lipopolymer significantly reduced the desorption rate of the ceramide from the liposome bilayer, thereby allowing liposomes containing C6Cer to reach the tumor site and to demonstrate therapeutic efficacy. PMID- 18512884 TI - Application of mean field boundary potentials in simulations of lipid vesicles. AB - A method is presented to enhance the efficiency of simulations of lipid vesicles. The method increases computational speed by eliminating water molecules that either surround the vesicle or reside in the interior of the vesicle, without altering the properties of the water at the membrane interface. Specifically, mean field force approximation (MFFA) boundary potentials are used to replace both the internal and external excess bulk solvent. In addition to reducing the cost of simulating preformed vesicles, the molding effect of the boundary potentials also enhances the formation and equilibration of vesicles from random solutions of lipid in water. Vesicles with diameters in the range from 20 to 60 nm were obtained on a nanosecond time scale, without any noticeable effect of the boundary potentials on their structure. PMID- 18512885 TI - Stable hairpins with beta-peptides: route to tackle protein-protein interactions. AB - Experimental and theoretical data demonstrate that sequences of heterochiral beta (2,3)-amino acids and a turn-inducing beta-dipeptide adopt hairpin-like structures in methanol. On the basis of extensive canonical and replica exchange MD simulations, we could transfer these findings to water as the solvent of physiological relevance. We show that rationally designed beta-peptides exhibit a higher folding tendency and a more robust hairpin structure formation in water compared with alpha-peptides. Furthermore, our designed scaffold enables the addition of a wide variety of functions without disrupting the structure. Since hairpins are often involved in protein interactions, the very stable hairpin-like fold of our designed beta-peptides might be used as a lead scaffold for the design of molecules that specifically modulate protein-protein interactions. This is demonstrated by application of this concept to the recognition of proline-rich sequences (PRS) by WW domains, an important interaction in cell signaling. We focus on the possibility to imitate the strands 2 and 3 of any WW domain as a minimal motif to recognize their target sequences PPXY. We conclude that rationally designed beta-peptide hairpins can serve as scaffolds not only to tackle PPII recognition but also to open up a way to influence a wide variety of protein-protein interactions. PMID- 18512886 TI - Brownian dynamics simulations of single-wall carbon nanotube separation by type using dielectrophoresis. AB - We theoretically investigate the separation of individualized metallic and semiconducting single-wall carbon nanotubes (SWNTs) in a dielectrophoretic (DEP) flow device. The SWNT motion is simulated by a Brownian dynamics (BD) algorithm, which includes the translational and rotational effects of hydrodynamic, Brownian, dielectrophoretic, and electrophoretic forces. The device geometry is chosen to be a coaxial cylinder because it yields effective flow throughput, the DEP and flow fields are orthogonal to each other, and all the fields can be described analytically everywhere. We construct a flow-DEP phase map showing different regimes, depending on the relative magnitudes of the forces in play. The BD code is combined with an optimization algorithm that searches for the conditions that maximize the separation performance. The optimization results show that a 99% sorting performance can be achieved with typical SWNT parameters by operating in a region of the phase map where the metallic SWNTs completely orient with the field, whereas the semiconducting SWNTs partially flow-align. PMID- 18512887 TI - Ultrathin-film growth of para-sexiphenyl (I): submonolayer thin-film growth as a function of the substrate temperature. AB - The para-sexiphenyl (p-6P) monolayer film induces weak epitaxy growth (WEG) of disk-like organic semiconductors, and their charge mobilities are increased dramatically to the level of the corresponding single crystals [Wang et al., Adv. Mater. 2007, 19, 2168]. The growth behavior and morphology of p-6P monolayer film play decisive roles on WEG. Here, we investigated the growth behavior of p-6P submonolayer film as a function of the substrate temperature. Its growth exhibited two different mechanisms at high and low substrate temperature. At high substrate temperature (>60 degrees C), the mechanism of diffusion-limited aggregation controlled the growth of submonolayer thin film with fractal islands, whereas at low substrate temperature (< or =60 degrees C), the submonolayer thin film was composed of the compact islands. Its growth exhibited another growth mechanism in which the stable compact islands were formed by dissociation and reorganization of the metastable disordered film. The substrate temperature of about 180 degrees C may be optimal to fabricate high-quality p-6P monolayer film with large-size domains and low saturated island density of about 0.018 microm( 2). PMID- 18512888 TI - Temperature study of a glycine radical in the solid state adopting a DFT periodic approach: vibrational analysis and comparison with EPR experiments. AB - The major radiation-induced radical in crystalline glycine is examined using DFT calculations, in which both molecular environment and temperature are accounted for. This is achieved by molecular dynamics simulations of the radical embedded in a supercell under periodic boundary conditions. At 100 and 300 K, a vibrational analysis is performed based on Fourier transformation of the atomic velocity autocorrelation functions. By the use of a novel band-pass filtering approach, several vibrational modes are identified and associated with experimental infrared and Raman assignments. Decomposition of the calculated spectra in terms of radical motion reveals that several vibrational modes are unique to the radical, the most prominent one at 702 cm(-1) corresponding to out of-plane motion of the paramagnetic center, inversely coupled with similar motion of the carboxyl carbon. A hybrid periodic/cluster scheme is used to evaluate the EPR properties of the glycine radical along the MD trajectories resulting in temperature dependent magnetic properties. These are compared with available experimental data conducted at 77 K and room temperature. Ground state or low temperature calculations yield very good agreement with 77 K experimental EPR properties. From the 300 K simulations, an important improvement is achieved on the isotropic hyperfine coupling of the (13)C tensor, which becomes closer to the value measured at room temperature. It is established that this is the result of a nonlinear relation between the planarity of the radical center and the isotropic couplings of the nuclei bound to it. Finally, a critical reevaluation of the experimental (14)N hyperfine tensor data strongly suggests that an erroneous tensor was reported in literature. It is convincingly shown that from the same experimental data set a different tensor can be derived, which is in substantially better agreement with all calculations. PMID- 18512889 TI - Interfacial assemblies of cyanine dyes and gemini amphiphiles with rigid spacers: regulation and interconversion of the aggregates. AB - A series of gemini amphiphiles with a pyridinium headgroup and rigid spacers were designed, and their interfacial assemblies with cyanine dyes, 3,3-disulfopropyl-9 methyl-selenacarbocyanine (SeCy) and 3,3-disulfopropyl-4,5,4',5'-dibenzo-9-methyl thiocarbocyanine (MTC), through the air/water interface were investigated. Although the dyes have intrinsic tendencies to form J- or H-aggregates in aqueous solutions, their aggregation could be regulated in the complex films. Depending on the spacer, both J- and H-aggregates were formed for SeCy, whereas an H aggregate with different absorption bands was obtained for MTC. Furthermore, the formed aggregates in the films could be reversibly switched by treating the complex films with HCl, NH3 gas, or water vapor. The J-aggregates of SeCy could be changed into H-aggregates and recovered through an alternative HCl/NH3 exposure in the films. The H-aggregate of the MTC film could be changed into J aggregate upon exposure to HCl gas and subsequently put into air or NH 3 gas, and the thus-formed J-aggregate could be changed into H-aggregate under hot water vapor. In addition, such a reversible switch between different aggregates of MTC was only observed when the gemini amphiphiles with rigid spacers were applied. A possible explanation related to the protonation of the dye and the reorganizations in the film during the interconversion between different aggregates was proposed. PMID- 18512890 TI - Ab initio molecular dynamics simulation of a 1-ethyl-3-methylimidazolium fluoride hydrogen fluoride mixture. AB - The intermolecular structure and dynamics of an acidic 1-ethyl-3 methylimidazolium fluoride-hydrogen fluoride solution ([emim][F].2.3HF) have been studied through a Car-Parrinello molecular dynamics simulation. The calculated structure factor is found to be in good agreement with X-ray scattering data. The solution consists of [emim] cations and polyfluoride anions of the kind F(HF)n( ). With increasing n, the length of the H-F covalent bond in the polyfluoride species is found to decrease, with a concomitant blue shift in the frequency of its stretching mode. Evidence for the presence of a hydrogen bond between the acidic ring hydrogen of the cation and the fluoride ion is presented. PMID- 18512891 TI - Structured water and water-polymer interactions in hydrogels of molecularly imprinted polymers. AB - Recently, molecular imprinting technology has fleetly developed for applications in different fields. It shows great potential in sensor design, drug delivery, chromatography separation, catalysis, chiral synthesis, and especially in the molecular recognition field. In this work, a cubic model of a hydrogel network was developed and an infinite hydrogel backbone network was constructed for molecular dynamics simulation. The water structure and water-polymer interaction was investigated from the radial distribution function and the viewpoint of the hydrogen-bonding system. It is found that the hydrogen bonds between polymer and water strongly depress the diffusion of water molecules and enhance the structure of water in the system. The greater the network mesh size of the polymer, the weaker the structure of the water. The decreasing of the density of hydrogen bonds between polymer and water is the major factor that leads to the weakening of water structure. PMID- 18512892 TI - Rupture force of single supramolecular bonds in associative polymers by AFM at fixed loading rates. AB - Atomic-force-microscopy-based single-molecule force spectroscopy (AFM-SMFS) was used to study the bond strength of self-complementary hydrogen-bonded complexes based on the 2-ureido-4[1H]-pyrimidinone (UPy) quadruple H-bond motif in hexadecane (HD). The unbinding force corresponding to single UPy-UPy dimers was investigated at a fixed piezo retraction rate in the nonequilibrium loading rate regime. The rupture force of bridging supramolecular polymer chains formed between UPy-functionalized substrates and AFM tips in the presence of a bis-UPy derivative was found to decrease with increasing rupture length. The rupture length was identified as the chain length of single, associating polymers, which allowed us to determine the number of supramolecular bonds (N) at rupture. The rupture force observed as a function of N was in quantitative agreement with the theory on uncooperative bond rupture for supramolecular linkages switched in a series. Hence, the value of the dimer equilibrium constant Keq=(1.3+/-0.5) x 10(9) M(-1), which is in good agreement with previously estimated values, was obtained by SMFS of supramolecular polymers at a single loading rate. PMID- 18512893 TI - The primary electron donor of photosystem II of the cyanobacterium Acaryochloris marina is a chlorophyll d and the water oxidation is driven by a chlorophyll a/chlorophyll d heterodimer. AB - We present a theoretical analysis of the flash-induced absorbance difference spectrum assigned to the formation of the secondary radical pair P(+)QA(-) in photosystem II of the chlorophyll d-containing cyanobacterium Acaryochloris marina. An exciton Hamiltonian determined previously for chlorophyll a-containing photosystem II complexes is modified to take into account the occupancy of certain binding sites by chlorophyll d instead of chlorophyll a. Different assignments of the reaction center pigments to chlorophyll a or d from the literature are investigated in the calculation of the absorbance difference spectrum. A quantitative explanation of the experimental data requires one chlorophyll a molecule per reaction center, located at the site of P(D1). The remaining sites are occupied by chlorophyll d and pheophytin a. By far, the lowest site energy is found for the accessory chlorophyll of the D1 branch, Chl(D1), which represents the sink of excitation energy and therefore the primary electron donor. The cationic state is stabilized at the chlorophyll a, which drives the oxidation of water. PMID- 18512894 TI - Multiple interactions in the self-association of porphyrin discotic mesogens. AB - The conformational preferences and the self-associational behaviors of two hemin derived porphyrin compounds, a tetramethyl ester and a liquid crystalline tetrakis(3,5-didodecyloxyphenyl)ester, have been studied by UV/vis and (1)H NMR spectroscopy in solution. Results indicate that the 3,5-didodecyloxyphenyl units play an important role in both the conformational and the self-associational behaviors of the mesomorphic tetraester. In the monomeric, nonassociated species, the two propionic 3,5-didodecyloxyphenyl esters establish mutual CH/pi interactions that restrict the fluctuative behavior of the chains. In the dimeric, self-associated species, intermolecular CH/pi interactions occur in addition to the pi-pi stacking of the porphyrin cores. The temperature-dependent addition of side CH/pi interactions to the pi-pi stacking of the porphyrin rings accounts for the observed tightening and for the slower dynamics of the dimeric structure. The relationship between the self-associational behavior and the mesomorphism of the hemin-derived porphyrin tetraesters is also discussed. PMID- 18512895 TI - Prediction of the equilibrium structures and photomagnetic properties of the Prussian blue analogue RbMn[Fe(CN)6] by density functional theory. AB - A periodic density functional theory method using the B3LYP hybrid exchange correlation potential is applied to the Prussian blue analogue RbMn[Fe(CN)6] to evaluate the suitability of the method for studying, and predicting, the photomagnetic behavior of Prussian blue analogues and related materials. The method allows correct description of the equilibrium structures of the different electronic configurations with regard to the cell parameters and bond distances. In agreement with the experimental data, the calculations have shown that the low temperature phase (LT; Fe(2+)(t(6)2g, S = 0)-CN-Mn(3+)(t(3)2g e(1)g, S = 2)) is the stable phase at low temperature instead of the high-temperature phase (HT; Fe(3+)(t(5)2g, S = 1/2)-CN-Mn(2+)(t(3)2g e(2)g, S = 5/2)). Additionally, the method gives an estimation for the enthalpy difference (HT <--> LT) with a value of 143 J mol(-1) K(-1). The comparison of our calculations with experimental data from the literature and from our calorimetric and X-ray photoelectron spectroscopy measurements on the Rb0.97Mn[Fe(CN)6]0.98 x 1.03 H2O compound is analyzed, and in general, a satisfactory agreement is obtained. The method also predicts the metastable nature of the electronic configuration of the high temperature phase, a necessary condition to photoinduce that phase at low temperatures. It gives a photoactivation energy of 2.36 eV, which is in agreement with photoinduced demagnetization produced by a green laser. PMID- 18512896 TI - Theoretical investigation of the Fe+-catalyzed oxidation of acetylene by N2O. AB - The gas-phase Fe(+)-mediated oxidation of acetylene by N2O on both sextet and quartet potential energy surfaces (PESs) is theoretically investigated using density functional theory. Geometries and energies of all the stationary points involved in the catalytic reaction are located. For the catalytic cycles, the crucial step is the initial N2O reduction by Fe(+) to form FeO(+), in which a direct O-abstraction mechanism is located on the sextet PES, whereas the quartet pathway favors a N-O insertion mechanism. Spin inversion moves the energy barrier for this process downward to a position below the ground-state entrance channel. The second step of the catalytic cycles involves two mechanisms corresponding to direct hydrogen abstraction and cyclization. The former mechanism accounts for the ethynol formation with the upmost activation barrier below the entrance channel by about 5 kcal/mol. The other mechanism involves a "metallaoxacyclobutene" structure, followed by four possible pathways, i.e., direct dissociation, C-C insertion, C-to-O hydrogen shift, and/or C-to-C hydrogen shift. Among these pathways, strong exothermicities as well as energetically low location of the intermediates suggest oxidation to ketene and carbon monoxide along the C-to-C hydrogen shift pathway is the most favorable. Reduction of the CO loss partner FeCH2(+) by another N2O molecule constitutes the third step of the catalytic cycles, which contains direct abstraction of O from N2O giving OFeCH2(+), intramolecular rearrangement to form Fe(+)-OCH2, and nonreactive dissociation. This reaction is also energetically favored considering the energy acquired from the initial reactants. PMID- 18512897 TI - Time-resolved cavity ringdown measurements and kinetic modeling of the pressure dependences of the recombination reactions of SiH2 with the alkenes C2H4, C3H6, and t-C4H8. AB - Room-temperature rate constants for the pressure-dependent reactions SiH2 + ethene, propene, and t-butene have been determined at total pressures of 3.3 mbar < or = p < or = 300 mbar with Ar as buffer gas. SiH2 was detected by means of time-resolved cavity ringdown spectroscopy (CRDS), and the deconvolution of ringdown, kinetics, and laser bandwidth effect was accomplished with the extended simultaneous kinetics and ringdown model (eSKaR). In this way, pseudofirst-order rate constants could be extracted from nonexponential ringdown profiles. The recombination reactions, including the reaction SiH2 + i-butene, have been modeled based on the simplified statistical adiabatic channel model (SACM) and weak collision energy-grained master equation (ME) simulations. The influence of an interfering fast isomerization channel was investigated based on the Rice, Ramsperger, Kassel, Marcus theory (RRKM) and was found to be only important for the C2H4 reaction. Using ab initio energies (G3) and structures (MP2/6-311G(d,p)) as input parameters for the kinetic models, a consistent description of the pressure and temperature dependences of all four reactions was possible. In the temperature range 295 K < or = T < or = 600 K, the extrapolated limiting high pressure rate constants, k(infinity)(C2H4)/(cm(3) x mol(-1) x s(-1)) = 1.9 x 10(14) (T/K)(-0.065), k(infinity)(C3H6)/(cm(3) x mol(-1) x s(-1)) = 1.3 x 10(14) (T/K)(0.075), k(infinity)(i-C4H8) = 1.8 x 10(14) cm(3) x mol(-1) x s(-1), and k(infinity)(t-C4H8)/(cm(3) x mol(-1) x s(-1)) = 4.6 x 10(13) (T/K)(0.21), are close to the collision number and are more or less temperature independent. In the case of ethene, probably due to the approximate treatment of rotational effects and/or the interfering isomerization process, the applied model slightly underestimates the falloff and thus yields too high extrapolated rate constants at p < 10 mbar. PMID- 18512898 TI - Infrared matrix isolation study of the thermal and photochemical reactions of ozone with dimethylzinc. AB - The matrix isolation technique has been combined with infrared spectroscopy and theoretical calculations to explore the reaction of (CH3)2Zn with O3 over a range of time scales. Upon twin jet deposition, an initial cage pair complex was observed, along with formation of the novel H3COZnCH3 species. Subsequent UV irradiation destroyed the complex and greatly increased the yield of H3COZnCH3. An extensive set of bands were seen for this molecule, and (18)O spectroscopic data were obtained as well. The identification of this species was supported by theoretical calculations at the B3LYP/6-311++g(d,2p) level. Merged jet deposition led to a very different set of products, including H2CO, CH3OH and C2H6, identifications that were confirmed by (18)O substitution. In addition, a new variable length concentric deposition technique was developed to permit study of the time scales between twin (relatively short) and merged (relatively longer) reaction times. Mechanistic inferences for this reaction are discussed. PMID- 18512899 TI - Quantitative distinction between competing intramolecular bond twisting and solvent relaxation dynamics: an ultrafast study. AB - Often an intramolecular relaxation process takes place in a time scale similar to that of the solvent relaxation process. Under these circumstances the dynamic Stokes' shift of the probe can be modulated by the combined effect of these two relaxation processes. In the present article we have studied ultrafast solvent relaxation using three different coumarin dyes and proposed a methodology for the quantitative separation of the dynamics of two competing processes, namely, solvent relaxation and bond twisting, that take place simultaneously in the present systems. PMID- 18512900 TI - 4-N-, 4-S-, and 4-O-chloroquine analogues: influence of side chain length and quinolyl nitrogen pKa on activity vs chloroquine resistant malaria. AB - Using predictions from heme-quinoline antimalarial complex structures, previous modifications of chloroquine (CQ), and hypotheses for chloroquine resistance (CQR), we synthesize and assay CQ analogues that test structure-function principles. We vary side chain length for both monoethyl and diethyl 4-N CQ derivatives. We alter the pKa of the quinolyl N by introducing alkylthio or alkoxy substituents into the 4 position and vary side chain length for these analogues. We introduce an additional titratable amino group to the side chain of 4-O analogues with promising CQR strain selectivity and increase activity while retaining selectivity. We solve atomic resolution structures for complexes formed between representative 4-N, 4-S, and 4-O derivatives vs mu-oxo dimeric heme, measure binding constants for monomeric vs dimeric heme, and quantify hemozoin (Hz) formation inhibition in vitro. The data provide additional insight for the design of CQ analogues with improved activity vs CQR malaria. PMID- 18512901 TI - Conformationally constrained analogues of N-(piperidinyl)-5-(4-chlorophenyl)-1 (2,4- dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141716): design, synthesis, computational analysis, and biological evaluations. AB - Structure-activity relationships (SARs) of 1 (SR141716) have been extensively documented, however, the conformational properties of this class have received less attention. In an attempt to better understand ligand conformations optimal for receptor recognition, we have designed and synthesized a number of derivatives of 1, including a four-carbon-bridged molecule (11), to constrain rotation of the diaryl rings. Computational analysis of 11 indicates approximately 20 kcal/mol energy barrier for rotation of the two aryl rings. NMR studies have determined the energy barrier to be approximately 18 kcal/mol and suggested atropisomers could exist. Receptor binding and functional studies with these compounds displayed reduced affinity and potency when compared to 1. This indicates that our structural modifications either constrain the ring systems in a suboptimal orientation for receptor interaction or the introduction of steric bulk leads to disfavored steric interactions with the receptor, and/or the relatively modest alterations in the molecular electrostatic potentials results in disfavored Coulombic interactions. PMID- 18512902 TI - Metal-organic framework with rationally tuned micropores for selective adsorption of water over methanol. AB - A microporous metal-organic framework 1, Cu(R-GLA-Me)(4,4'-Bipy) 0.5.0.55H 2O (R GLA-Me = R-2-methylglutarate, 4,4'-Bipy = 4,4'-bipyridine), with a primitive cubic net was synthesized and characterized. With pores of about 2.8 x 3.6 A, the activated 1a exhibits exclusive adsorption of water over methanol in a binary water-methanol (1:1) liquid mixture. PMID- 18512903 TI - Synthesis, structures, and magnetic properties of face-sharing heterodinuclear Ni(II)-Ln(III) (Ln = Eu, Gd, Tb, Dy) complexes. AB - Heterodinuclear [(Ni (II)L)Ln (III)(hfac) 2(EtOH)] (H 3L = 1,1,1 tris[(salicylideneamino)methyl]ethane; Ln = Eu, Gd, Tb, and Dy; hfac = hexafluoroacetylacetonate) complexes ( 1.Ln) were prepared by treating [Ni(H 1.5L)]Cl 0.5 ( 1) with [Ln(hfac) 3(H 2O) 2] and triethylamine in ethanol (1:1:1). All 1.Ln complexes ( 1.Eu, 1.Gd, 1.Tb, and 1.Dy) crystallized in the triclinic space group P1 (No. 2) with Z = 2 with very similar structures. Each complex is a face-sharing dinuclear molecule. The Ni (II) ion is coordinated by the L (3-) ligand in a N 3O 3 coordination sphere, and the three phenolate oxygen atoms coordinate to an Ln (III) ion as bridging atoms. The Ln (III) ion is eight coordinate, with four oxygen atoms of two hfac (-)'s, three phenolate oxygen atoms of L (3-), and one ethanol oxygen atom coordinated. Temperature-dependent magnetic susceptibility and field-dependent magnetization measurements showed a ferromagnetic interaction between Ni (II) and Gd (III) in 1.Gd. The Ni (II)-Ln (III) magnetic interactions in 1.Eu, 1.Tb, and 1.Dy were evaluated by comparing their magnetic susceptibilities with those of the isostructural Zn (II)-Ln (III) complexes, [(ZnL)Ln(hfac) 2(EtOH)] ( 2.Ln) containing a diamagnetic Zn (II) ion. A ferromagnetic interaction was indicated in 1.Tb and 1.Dy, while the interaction between Ni (II) and Eu (III) was negligible in 1.Eu. The magnetic behaviors of 1.Dy and 2.Dy were analyzed theoretically to give insight into the sublevel structures of the Dy (III) ion and its coupling with Ni (II). Frequency dependence in the ac susceptibility signals was observed in 1.Dy. PMID- 18512904 TI - Synthesis and structural determination of a hexanuclear zirconium glycine compound formed in aqueous solution. AB - Single crystals of a zirconium-glycine compound have been first grown in aqueous solution via an excess acid and extra stabilizer approach. The crystal structure [Zr 6(OH (-)) 8(H 2O) 8(HGly) 4(Gly (-)) 4].(SO 4 (2-)) 6.14H 2O ( CP-1) has been characterized by single-crystal X-ray diffraction. The structure revealed that it is composed of hexa-zirconium octahedral clusters coordinated by eight carboxylic acid groups of glycine. The charge assignment is also consistent with the electronic structure calculations, and the computational result reveals that the Zr 6 core should have no skeleton electrons. PMID- 18512905 TI - Synthesis of new copper cyanide complexes via the transformation of organonitrile to inorganic cyanide. AB - Hydrothermal reaction of diaminomaleonitrile and copper salts under different conditions resulted in copper cyanide coordination polymers {[Cu(H 2O)(NH 3) 4][Cu 3(CN) 5].H 2O} n ( 1), {(CH 3) 4N[Cu(H 2O)(NH 3) 4][Cu 4(CN) 7]} n ( 2), and {(CH 3OH 2) 2[Cu 2(CN) 3]} n ( 3). 1 and 2 are new mixed-valence Cu(I,II), two 3D organic-inorganic molecular framework complexes that exhibit ionic inclusion. 3 is an open copper cyanide framework hosting a guest molecule. Cyanides in 1, 2, and 3 are produced by in situ C-C bond cleavage of diaminomaleonitrile, and then the remaining product is oxidized to form an oxalate group. The potential porosity of the hydrated coordination polymer 3 was estimated using a computational method based on Connolly's algorithm. PMID- 18512906 TI - Catalytic phenylborylation reaction by iridium(0) nanoparticles produced from hydridoiridium carborane. AB - Well-dispersed iridium(0) nanoparticles stabilized with the ionic liquid, trihexyltetradecylphosphonium methylsulfonate, [THTdP][MS], have been successfully prepared by reduction of the precursor hydridoiridium carborane, (Ph 3P) 2Ir(H)(7,8- nido-C 2B 9H 11). The iridium nanoparticles were found to be active catalysts for arylborylation, forming boric acids. The activity of the catalyst has been investigated as a function of the activating base, and reaction conditions. The highest yield of 91% was achieved in a microwave reactor using the base, tetra-2-pyridinylpyrazine, in the presence of [THTdP][MS]. The catalytic system could be recycled at least six times with less than a 0.5% loss of activity. PMID- 18512907 TI - Cooperative catalysis with chiral Bronsted acid-Rh2(OAc)4: highly enantioselective three-component reactions of diazo compounds with alcohols and imines. AB - An asymmetric three-component reaction of diazo compounds and alcohols with imines catalyzed cooperatively by a rhodium complex and a chiral Bronsted acid provides a general and efficient entry to beta-amino-alpha-hydroxyl acid derivatives in high yields with excellent stereoselectivities. PMID- 18512908 TI - Pristine simple oxides as visible light driven photocatalysts: highly efficient decomposition of organic compounds over platinum-loaded tungsten oxide. AB - Tungsten oxide loaded with nanoparticulate platinum is demonstrated to exhibit high activity for the decomposition of organic compounds both in liquid and gas phases; the activity was almost comparable to that of TiO2 under UV light irradiation and much higher than that of nitrogen-doped TiO2 under visible irradiation. PMID- 18512909 TI - Tunneling in C-H oxidation reactions by an oxoiron(IV) porphyrin radical cation: direct measurements of very large H/D kinetic isotope effects. AB - Rate constants for oxidations of benzyl alcohol-d0 and -d7 by oxoiron(IV) tetramesitylporphyrin radical cation perchlorate in acetonitrile were measured in single turnover kinetic studies. The kinetic isotope effect (kH/kD) increased from 28 at 23 degrees C to 360 at -30 degrees C due to extensive hydrogen atom tunneling that was analyzed in terms of a parabolic energy barrier to tunneling. Similarly, large KIE values were found for oxidations of ethylbenzene-d0 and -d10 at room temperature. The large KIE values are a function of the porphyrin identity, and porphyrins containing electron-withdrawing groups display normal KIEs. KIEs found under catalytic turnover conditions are somewhat smaller than those obtained in single turnover reactions. The results should serve as benchmarks for computational studies of C-H oxidations by porphyrin and heme-iron oxo systems. PMID- 18512910 TI - Simultaneous detection and deconvolution of congested NMR spectra containing three isotopically labeled species. AB - We present a procedure for isolating subspectra corresponding to individual protein or peptide components in a ternary mixture or complex. Each of the three component species is labeled differently: species A uniformly with 15N, species B uniformly with 15N and 13C, and species C uniformly with 15N but selectively with 13C' or 13Calpha. By using the dual carbon label selective HSQC (DCLS-HSQC) pulse sequence and exploiting differences in 1J 15N-13C coupling patterns to filter selected 15N resonances from detection during a constant time period, a subspectrum from each species can be generated from three spectra acquired from a single sample. Many important biological pathways involve dynamic interactions among members of multicomponent protein assemblies, and this approach offers a powerful way to monitor such processes. PMID- 18512911 TI - A fluorescent core-shell dendritic macromolecule specifically stains the extracellular matrix. AB - The extracellular cell matrix (ECM) surrounds cells and plays important roles in many aspects of cellular fate, including cell migration, stem cell differentiation, and cancer progression. So far, there is no fluorescent dye to directly visualize the ECM network. Here we present a positively charged fluorescent core-shell dendritic macromolecule containing multiple -NH2 groups which specifically binds to highly negatively charged ECM components. Due to its advantageous optical properties and biological specificity, the dye is useful as a routine tool to label the ECM in life science research. PMID- 18512912 TI - Cu-facilitated C-O bond formation using N-hydroxyphthalimide: efficient and selective functionalization of benzyl and allylic C-H bonds. AB - A highly efficient protocol for the benzyl or allylic C-H functionalization of simple hydrocarbons has been developed using stoichiometric amounts of N hydroxyphthalimide and PhI(OAc)2 in the presence of CuCl catalyst. The reaction was revealed to proceed via a radical pathway, in which phthalimide N-oxyl (PINO) radical plays a dual role, serving as a catalytic hydrogen abstractor from hydrocarbons as well as a stoichiometric reagent to couple with the resultant alkyl radicals. PMID- 18512913 TI - Crystal structure of thioflavin T bound to the peripheral site of Torpedo californica acetylcholinesterase reveals how thioflavin T acts as a sensitive fluorescent reporter of ligand binding to the acylation site. AB - Acetylcholinesterase plays a key role in cholinergic synaptic transmission by hydrolyzing the neurotransmitter acetylcholine with one of the highest known catalytic rate constants. Hydrolysis occurs in a narrow and deep gorge that contains two sites of ligand binding: A peripheral site, or P-site, near the gorge entrance that contributes to catalytic efficiency both by transiently trapping substrate molecules as they enter the gorge and by allosterically accelerating the transfer of the substrate acyl group to a serine hydroxyl in an acylation site or A-site at the base of the gorge. Thioflavin T is a useful reporter of ligand interactions with the A-site. It binds specifically to the P site with fluorescence that is enhanced approximately 1000-fold over that of unbound thioflavin T, and the enhanced fluorescence is quenched 1.5- to 4-fold when another ligand binds to the A-site in a ternary complex. To clarify the structural basis of this advantageous signal change, we here report the X-ray structure of the complex of thioflavin T with Torpedo californica acetylcholinesterase. The two aromatic rings in thioflavin T are coplanar and are packed snugly parallel to the aromatic side chains of Trp279, Tyr334, and Phe330. Overlays of this structure with the crystal structures of Torpedo californica acetylcholinesterase complexes with either edrophonium or m-( N, N, N trimethylammonio)-2,2,2-trifluoroacetophenone, two small aromatic ligands that bind specifically to the A-site, indicate that the phenyl side chain of Phe330 must rotate to sterically accommodate both thioflavin T and the A-site ligand in the ternary complex. This rotation may allow some relaxation of the strict coplanarity of the aromatic rings in the bound thioflavin T and result in partial quenching of its fluorescence. PMID- 18512914 TI - Effect of thioxopeptide bonds on alpha-helix structure and stability. AB - Thioxoamide (thioamide) bonds are nearly isosteric substitutions for amides but have altered hydrogen-bonding and photophysical properties. They are thus well suited backbone modifications for physicochemical studies on peptides and proteins. The effect of thioxoamides on protein structure and stability has not been subject to detailed experimental investigations up to date. We used alanine based model peptides to test the influence of single thioxoamide bonds on alpha helix structure and stability. The results from circular dichroism measurements show that thioxoamides are strongly helix-destabilizing. The effect of an oxo-to thioxoamide backbone substitution is of similar magnitude as an alanine-to glycine substitution resulting in a helix destabilization of about 7 kJ/mol. NMR characterization of a helical peptide with a thioxopeptide bond near the N terminus indicates that the thioxopeptide moiety is tolerated in helical structures. The thioxoamide group is engaged in an i, i+4 hydrogen bond, arguing against the formation of a 3(10)-helical structure as suggested for the N-termini of alpha-helices in general and for thioxopeptides in particular. PMID- 18512915 TI - Are the radical centers in peptide radical cations mobile? The generation, tautomerism, and dissociation of isomeric alpha-carbon-centered triglycine radical cations in the gas phase. AB - The mobility of the radical center in three isomeric triglycine radical cations[G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) has been investigated theoretically via density functional theory (DFT) and experimentally via tandem mass spectrometry. These radical cations were generated by collision-induced dissociations (CIDs) of Cu(II)-containing ternary complexes that contain the tripeptides YGG, GYG, and GGY, respectively (G and Y are the glycine and tyrosine residues, respectively). Dissociative electron transfer within the complexes led to observation of [Y(*)GG](+), [GY(*)G](+), and [GGY(*)](+); CID resulted in cleavage of the tyrosine side chain as p-quinomethide, yielding [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+), respectively. Interconversions between these isomeric triglycine radical cations have relatively high barriers (> or = 44.7 kcal/mol), in support of the thesis that isomerically pure [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) can be experimentally produced. This is to be contrasted with barriers < 17 kcal/mol that were encountered in the tautomerism of protonated triglycine [Rodriquez C. F. et al. J. Am. Chem. Soc. 2001, 123, 3006-3012]. The CID spectra of [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) were substantially different, providing experimental proof that initially these ions have distinct structures. DFT calculations showed that direct dissociations are competitive with interconversions followed by dissociation. PMID- 18512916 TI - Microwave-induced multiple functionalization of carbon nanotubes. AB - We describe a new synthetic strategy to produce multifunctionalized carbon nanotubes using a combination of two different addition reactions, the 1,3 dipolar cycloaddition of azomethine ylides and the addition of diazonium salts, both via a simple and fast microwave-induced method. The presence of multifunctionality on the SWNTs has been confirmed using the most useful techniques for the characterization of carbon nanotubes. The doubly functionalized SWNTs can be considered potentially useful for many interesting applications. PMID- 18512917 TI - Antiparallel arrangement of the helices of vesicle-bound alpha-synuclein. AB - alpha-Synuclein (alphaS) is the main component of Lewy bodies from Parkinson's disease. That alphaS binds to membranes is known, but the conformation it adopts is still unclear. Pulsed EPR on doubly spin-labeled variants of alphaS sheds light on the most likely structure. For alphaS bound to vesicles large enough to accommodate also the extended conformation, an antiparallel helix conformation is found. This suggests that the bent structure shown is the preferred conformation of alphaS on membranes. PMID- 18512918 TI - Exciton energy transfer-assisted photoluminescence brightening from freestanding single-walled carbon nanotube bundles. AB - Photoluminescence (PL) brightening is clearly observed through the direct morphology transition from isolated to thin bundled vertically- and individually freestanding single-walled carbon nanotubes (SWNTs). On the basis of the precise spectra analysis and equation-based estimation of the PL time trace, the origin of the PL brightening is consistently explained in terms of the exciton energy transfer through the tube bundles. The PL brightening is also revealed to obviously depend on SWNT diameters. Only the small-diameter rich sample can realize the PL brightening, which can be explained by the different concentrations of metallic SWNTs causing a PL quenching. Since it can be possible to fabricate brightly illuminating nanotubes on various kinds of substrates, the bundle engineering with freestanding nanotubes is expected to be a potential candidate for realizing the nanotube-based PL device fabrication. PMID- 18512919 TI - Calculation of residual dipolar couplings from disordered state ensembles using local alignment. AB - Residual dipolar couplings (RDCs) have been observed in disordered states of several proteins. While their nonuniform values were initially surprising, it has been shown that reasonable approximation of experimental RDCs can be obtained using simple statistical coil models and assuming global alignment of each structure, provided that many thousands of conformers are averaged. Here we show that, by using short local alignment tensors, we can achieve good agreement between experimental and simulated RDCs with far fewer structures than required when using global alignment. This makes the possibility of using RDCs as direct restraints in structural calculations of disordered proteins much more feasible. In addition, it provides insight into the nature of RDCs in disordered states, suggesting that they are primarily reporting on local structure. PMID- 18512920 TI - Synchrotron X-ray charge density study of coordination polymer Co3(C8H4O4)4(C4H12N)2(C5H11NO)3 at 16 K. AB - The charge density (CD) of coordination polymer Co3(C8H4O4)4(C4H12N)2(C5H11NO)3 (1) has been determined from multipole modeling of structure factors obtained from single-crystal synchrotron X-ray diffraction measurements at 16 K. The crystal structure formally contains a negatively charged framework with cations and neutral molecules in the voids. However, the CD suggests that the framework is close to neutral, and therefore qualitative conclusions based on formal charge counting, e.g., about guest inclusion properties, will be incorrect. There are considerable differences in the charge distributions of the three unique benzenedicarboxylic acid linkers, which are widely used in coordination polymers. This suggests that the electrostatic properties of coordination polymer cavities, and thereby their inclusion properties, are highly tunable. The electron density topology shows that the tetrahedrally coordinated Co atom has an atomic volume which is 15% larger than that of the octahedrally coordinated Co atom. The crystal structure has both ferromagnetic and antiferromagnetic interactions, but no direct metal-metal bonding is evidenced in the CD. The magnetic ordering therefore takes place through superexchange in the oxygen bridges and the aromatic linkers. Bonding analysis of the experimental CD reveals that two oxygen atoms, O(1) and O(11), have significant covalent contributions to the metal ligand bonding, whereas all other oxygen atoms have closed-shell interactions with the metals. This indicates that these two oxygen atoms are the key mediators of the magnetic ordering. PMID- 18512921 TI - Broadly hysteretic H2 adsorption in the microporous metal-organic framework Co(1,4-benzenedipyrazolate). AB - Reaction of Co(CF3SO3)2 with the new molecule 1,4-benzenedi(4'-pyrazolyl) (H2BDP) in N,N'-diethylformamide (DEF) at 130 degrees C generates the metal-organic framework Co(BDP).2DEF.H2O (1). X-ray analysis reveals the structure of 1 to contain chains of tetrahedrally ligated Co2+ ions linked through BDP2- ligands to generate a three-dimensional framework with 10 x 10 A2 channels. Thermogravimetric data shows the framework to have a high thermal stability, and complete desolvation occurs upon heating at 170 degrees C under dynamic vacuum for two days to afford 1d. X-ray powder diffraction data indicates that 1d possesses a substantially different structure, but converts back to 1 upon exposure to DEF, consistent with the presence of a flexible framework. Nitrogen adsorption isotherms measured for 1d at 77 and 87 K reveal an unprecedented five step adsorption process and a Langmuir surface area of 2670 m2/g. In addition, high-pressure H2 adsorption data reveal hysteretic uptake and release, with hysteresis loops of width 1.1, 3.8, 13, and 27 bar that shift to higher pressures as the temperature increases from 50 to 65, 77, and 87 K, respectively. The high H2 uptake capacity of 5.5 excess wt % at 50 K suggests that such materials could potentially find utility for hydrogen storage via a kinetic trapping mechanism. Variable-temperature kinetics measurements have also allowed the first study of H2 diffusion within a metal-organic framework, revealing an energy barrier of 0.62 kJ/mol for H2 diffusing within the pores. PMID- 18512922 TI - Engineered biosynthesis of antiprotealide and other unnatural salinosporamide proteasome inhibitors. AB - A new shunt in the phenylalanine biosynthetic pathway to the nonproteinogenic amino acid L-3-cyclohex-2'-enylalanine was exploited in the marine bacterium Salinispora tropica by mutagenesis to allow for the genetic engineering of unnatural derivatives of the potent proteasome inhibitor salinosporamide A (2) such as antiprotealide (1). PMID- 18512923 TI - Iridium corroles. AB - This work reports the synthesis and full characterization of 5,10,15-tris pentafluorophenylcorrolato-iridium(III) bis-trimethylamine 1 and its octabromo derivative 2. The corrole is planar in both cases (the mean deviation from the plane is as low as 0.0371 A for 1 and 0.0325 A for 2), the UV-vis spectra display a split Soret band with a shoulder attributable to an MLCT transition, and cyclic voltammetry reveals that the iridium(II) oxidation state cannot be accessed, while the oxidation to formal iridium(IV) complexes is achieved at much lower potentials than in other coordination environments. PMID- 18512924 TI - Cell-based fluorescence screen for K+ channels and transporters using an extracellular triazacryptand-based K+ sensor. AB - K+ channels and K+-coupled membrane transporters are important targets for drug discovery. We previously developed a triazacryptand (TAC)-based K+ sensor, TAC Red, and demonstrated its utility to image K+ waves in mouse brain in vivo (Padmawar et al. Nat. Methods. 2005, 2, 825-827). Here, we synthesized a green fluorescing dextran conjugate of TAC-bodipy ("TAC-Limedex") for use as an extracellular K+ sensor and demonstrated its utility in measuring K+ transport across cell membranes. TAC-Limedex fluorescence increased by 50% with increasing [K+] from 0 to 2 mM and was insensitive to [Na+], [Cl-], or pH. K+ efflux from cells was quantified from increasing extracellular TAC-Limedex fluorescence following cell immersion in K+-free buffer. In HT-29 cells, K+ efflux was 2.0 +/- 0.1 micromol/cm2/s, increasing 8-fold following K+ channel activation by ATP; the increase in K+ efflux was inhibited by a K+ channel blocker or by preventing cytoplasmic calcium elevation. Electroneutral K+/Cl- cotransport was demonstrated in SiHa cells, in which K+ efflux was increased 3-fold by hypotonic challenge; the increase in K+ efflux was fully inhibited by a K+/Cl- transport blocker. K+ efflux measurements were adapted to a commercial fluorescence platereader for automated screening. The fluorescence-based K+ transport assay largely replaces assays requiring radioactive rubidium and is suitable for high-throughput identification of K+ transport modulators. PMID- 18512925 TI - Binary superlattices of PbSe and CdSe nanocrystals. AB - In this paper we show that self-organization of colloidal PbSe and CdSe semiconductor nanocrystals with a size ratio of 0.57 leads to binary structures with a AB2 or a cuboctahedral AB13 lattice. The type of superlattice formed can be regulated by the relative concentration of both nanocrystals in the suspension. PMID- 18512926 TI - The putative Diels-Alderase macrophomate synthase is an efficient aldolase. AB - We find that the putative Diels-Alderase macrophomate synthase (MPS) catalyzes the addition of pyruvate enolate, generated by decarboxylation of oxaloacetate, to a variety of aldehydes. Alkyl, aryl, and heteroaryl aldehydes are accepted as substrates, providing gamma-hydroxy-alpha-ketoacids in 35-95% yield with modest levels of stereochemical control. These aldol products, which are difficult to synthesize by other methods, are formed with efficiency comparable to that of macrophomate. Our results thus provide evidence that a two-step Michael-aldol pathway is a plausible alternative to the postulated [4 + 2] cycloaddition in the MPS-catalyzed addition of pyruvate enolate to 2-pyrones. They are also relevant to understanding the divergent evolution of type II pyruvate aldolases. PMID- 18512927 TI - Direct examination of H2O2 activation by a heme peroxidase. AB - Described here is the application of oxygen isotope fractionation together with computational methods, to elucidate a mechanism of enzymatic H2O2 activation. Horseradish peroxidase (HRP) has been the subject of intensive experimental and computational studies, yet questions remain as to the reversibility of the O-O cleavage step. New insight is afforded by the competitive oxygen kinetic isotope effect (18O KIE) upon H2O2 consumption determined under turnover conditions. The 18O KIE is compared to isotope effects calculated for the O-O heterolysis transition state and potential intermediates using density functional theory. In addition, experiments in enriched water provide evidence for HRP-catalyzed scrambling of the 18O label into the unreacted H2O2. The results provide an unprecedented view of H2O2 activation by a heme peroxidase and challenge the assumption of rate-limiting O-O heterolysis. PMID- 18512929 TI - Mild, efficient Friedel-Crafts acylations from carboxylic acids using cyanuric chloride and AlCl3. AB - A mild method for Friedel-Crafts acylation with aromatic and aliphatic carboxylic acids using cyanuric chloride, pyridine, and AlCl(3) was developed. Both inter- and intramolecular acylations were achieved at room temperature in high yield and in very short reaction times. PMID- 18512930 TI - Asymmetric synthesis of spiroketal, spiroether, and oxabicycle building blocks via stereoselective spiro- and bicycloannulation of 2-hydroxy dihydropyrans. AB - A modular asymmetric synthesis of spiroketal, spiroether, and oxabicycle building blocks is described based on the spiro- and bicycloannulation of alpha-hydroxy dihydropyrans, which were obtained from sulfoximine-substituted homoallylic alcohols. Key steps of the syntheses are stereoselective Ferrier-type O- and C glycosidation, ring-closing metathesis, and stereoselective Prins cyclization. PMID- 18512931 TI - Determination of proteins in refined and nonrefined oils. AB - Five methods using aqueous/organic solvents for the separation of proteins from oils were compared. The extraction with acetone-hexane followed by amino acid analysis was found to be the most suitable method for isolation and quantification of proteins from oils. The detection limit of the method was 0.18 mg protein/kg oil, and the quantification limit was 0.6 mg protein/kg. The relative repeatability limit for samples containing 1-5 mg protein/kg sample was 27%. The protein recovery ranged between 68 and 133%. Using this method, the protein content of 14 refined and nonrefined oils was determined. In none of the refined oils were proteins detected, whereas the protein content of the unrefined oils ranged between undetectable in extra virgin olive oil to 11 mg/kg in rapeseed oil. With sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with silver staining, many protein bands were visible in the unrefined soy, olive, peanut, and rapeseed oil samples. Proteins bands were not obtained from the refined fish oil. In the other refined oil samples, a few proteins bands could be visualized. Two protein bands with apparent molecular molecular masses of 58 and 64 kDa were always observed in these oils. PMID- 18512932 TI - Antiobese effects of novel saponins from edible seeds of Japanese horse chestnut (Aesculus turbinata BLUME) after treatment with wood ashes. AB - Recently, we have identified novel saponins from edible seeds of Japanese horse chestnut ( Aesculus turbinata BLUME) after processing the natural seeds with wood ashes to remove bitterness. We attempted to determine anti-obesity effects of those saponins from edible seeds as well as natural seeds. The purified individual components of saponins from natural and edible seeds inhibited pancreatic lipase in vitro. The potency was in the order of escins > desacylescins > deacetylescins. Escins Ib and IIb as well as deacetylescins Ib and IIb with the angeloyl moiety were more potent than the corresponding Ia and IIa series with the tigloyl moiety. Moreover, in vivo anti-obesity effects of the saponin fractions were monitored for 8 weeks in mice fed high-fat diets. Saponin fractions from both seeds significantly attenuated the elevation in body weight, the mass of peritoneal adipose tissues, and plasma triacylglycerol, which was accompanied by higher contents of undigested fats in feces without changes in food intake, indicating the effective inhibition of fat digestion in vivo. Taken together, saponin fractions including desacylescins and deacetylescins from edible seeds are potentially useful for the development of nutraceutical foods with anti-obesity effects and more attenuated bitter taste. PMID- 18512933 TI - Slowly digestible state of starch: mechanism of slow digestion property of gelatinized maize starch. AB - The mechanism underlying the previously reported parabolic relationship between amylopectin fine structure, represented by the weight ratio of linear short chains [degree of polymerization (DP < 13) to long chains (DP >/= 13], and slowly digestible starch (SDS) content was investigated from the viewpoint of starch retrogradation and substrate susceptibility to enzyme hydrolysis. A maize mutant sample, termed "highest long-chain starch" (HLCS) representing group I samples with a higher proportion of long chains, showed a bell-shaped SDS pattern with retrogradation time, whereas insignificant changes in SDS were found for the sample termed "highest short-chain starch" (HSCS) representing group II samples with a higher proportion of short chains. This corresponded to results from X-ray powder diffraction and differential scanning calorimetry that showed a rapid increase of crystallinity and enthalpy for HLCS during retrogradation, but negligible changes for sample HSCS. Therefore, retrogradation was associated with SDS content for group I samples, but not for group II samples. Analysis of amylopectin fine structure, SDS content, retrogradation enthalpy, SDS material debranching profile, and hydrolysis pattern demonstrated, for group I samples, that linear branched chains of DP 9-30 of amylopectin may act as anchor points to slow the digestion of branced-chain fractions of DP > 30, which constitute the major slowly digestible portion, whereas for group II samples, it is the inherent molecular structure of amylopectin with a higher amount of branches and shorter chains that is not favorable for rapid enzyme digestion. The concept of a slowly digestible starch state (SDS state) that could be a chemical or physical entity is proposed to better describe the mechanistic underpinning of the slow digestion property of starches. PMID- 18512934 TI - Identification of a powerful aroma compound in munster and camembert cheeses: ethyl 3-mercaptopropionate. AB - With the view to investigate the presence of thiols in cheese, the use of different methods of preparation and extraction with an organomercuric compound ( p-hydroxymercuribenzoate) enabled the isolation of a new compound. The analysis of cheese extracts by gas chromatography coupled with pulse flame photometry, mass spectrometry, and olfactometry detections led to the identification of ethyl 3-mercaptopropionate in Munster and Camembert cheeses. This compound, described at low concentrations as having pleasant, fruity, grapy, rhubarb, and empyreumatic characters, has previously been reported in wine and Concord grape but was never mentioned before in cheese. A possible route for the formation of this compound in relation with the catabolism of sulfur amino acids is proposed. PMID- 18512935 TI - Adsorption of Cry1Ab protein isolated from Bt transgenic rice on bentone, kaolin, humic acids, and soils. AB - Adsorption on a soil matrix of the insecticidal protein from Bacillus thuringiensis ( Bt) transgenic plants affects their accumulation and release and, hence, bioavailability in soil. Cry1Ab protein isolated from Bt transgenic rice was used to evaluate the adsorption and desorption on bentone, kaolin, and humic acids (HAs). The adsorption equilibrium of Cry1Ab protein was reached within 1-2 h for bentone and kaolin and within 4-8 h for HAs. The adsorption isotherms were better described by linear expressions ( R (2) >/= 0.973) rather than by the Freundlich model. No saturation was observed, even at the maximum concentration used (3.71 microg mL (-1)). The adsorbed protein was not easily desorbed at the used protein concentrations (0.18-3.71 microg mL (-1)); more than 50-70%, 70-80%, and 90% of the adsorbed protein remained on HAs, kaolin, and bentone, respectively, after washing with water. Adsorption and desorption of the Cry1Ab protein were further studied using five soils, and the isotherms were also well described by linear equations ( p < 0.05). Adsorption of the Cry1Ab protein on soils was positively related to the soil organic matter content. PMID- 18512937 TI - Detection of food additives by voltammetry at the liquid-liquid interface. AB - Electrochemistry at the liquid-liquid interface enables the detection of nonredoxactive species with electroanalytical techniques. In this work, the electrochemical behavior of two food additives, aspartame and acesulfame K, was investigated. Both ions were found to undergo ion-transfer voltammetry at the liquid-liquid interface. Differential pulse voltammetry was used for the preparation of calibration curves over the concentration range of 30-350 microM with a detection limit of 30 microM. The standard addition method was applied to the determination of their concentrations in food and beverage samples such as sweeteners and sugar-free beverages. Selective electrochemically modulated liquid liquid extraction of these species in both laboratory solutions and in beverage samples was also demonstrated. These results indicate the suitability of liquid liquid electrochemistry as an analytical approach in food analysis. PMID- 18512936 TI - Characterizing the metabolic fingerprint and anti-inflammatory activity of Hypericum gentianoides. AB - In this paper we characterize the metabolic fingerprint and first reported anti inflammatory activity of Hypericum gentianoides. H. gentianoides has a history of medical use by Native Americans, but it has been studied very little for biological activity. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analyses of a methanol extract show that H. gentianoides contains a family of over nine related compounds that have retention times, mass spectra, and a distinctive UV absorption spectra characteristic of certain acyl-phloroglucinols. These metabolites are abundant relative to other secondary products present in H. gentianoides, accounting for approximately 0.2 g per gram of dry plant tissue. H. gentianoides methanol extracts and a specific semipreparative HPLC fraction from these extracts containing the putative acyl-phloroglucinols reduce prostaglandin E 2 synthesis in mammalian macrophages. PMID- 18512938 TI - Grain qualities and their genetic derivation of 7 new rice for Africa (NERICA) varieties. AB - NERICA are interspecific rice varieties from crossing between the high-yielding Asian rice ( Oryza sativa spp. Japonica) with locally adapted African rice ( Oryza glaberrima). In this study, we analyzed grain qualities of 7 NERICA varieties (NERICA 1 to 7) and genetic derivation of quality-related genes. Quality analyses of NERICA grains showed that 7 NERICA varieties were clearly classified into two groups based on the difference of amylose content, and the difference influenced the pasting and physical properties of grains. Genetic analysis of the gene encoding granule-bound starch synthase I (GBSSI), which is known as a key enzyme on amylose synthesis in rice grain, revealed that varieties with higher amylose content ( approximately 29%) have the gene derived from O. glaberrima parent, and group 2 with lower amylose content ( approximately 22%) have the gene from O. sativa parent. These results indicated that the difference in amylose content as well as grain properties among 7 NERICA varieties is mainly determined by the genetic derivation of GBSSI. Further genetic analysis of starch synthesis-related genes suggested that the genetic derivation of SSIIa also influences the chain length of amylopectin in 7 NERICA varieties. PMID- 18512939 TI - Uptake of Zn and Fe by wheat (Triticum aestivum var. Greina) and transfer to the grains in the presence of chelating agents (ethylenediaminedisuccinic acid and ethylenediaminetetraacetic acid). AB - A way to decrease iron and zinc deficiency in humans is to biofortify foods by increasing the bioavailable contents in these elements. The aim of this work was to study if chelating agents could be used to increase the capture of Fe and Zn by wheat grains. Zn and/or Fe in combination with the chelating agents ethylenediaminedisuccinic acid (EDDS) or ethylenediaminetetraacetic acid (EDTA) were added at various times (i.e., at flower head formation, anthesis, and postanthesis) to spring wheat ( Triticum aestivum var. Greina) grown in nutrient solution. Treatments lasted for 2 weeks, and the plants were harvested at grain maturity. The shoots of treated plants accumulated higher Zn and/or Fe concentrations than untreated plants, depending on the treatment. The plants also accumulated significant concentrations of EDDS or EDTA in their shoots. Elevated Zn and Fe concentrations in the shoots did in most cases not lead to significantly higher Zn and Fe concentrations in the grains. The grains of plants treated with EDDS during flower head formation accumulated elevated Fe and Zn concentrations but at the cost of a reduction in yield. The control plants transferred higher percentages of Fe and Zn from the shoot into the grain than the treated plants. This indicates that EDTA and EDDS inhibited in most cases the translocation of Fe and Zn from the shoots into the grains. The amounts of EDDS and EDTA found in the grains of treated plants were very small. This indicates that there was little transfer of the chelates into the symplast and that the apoplastic pathway, which is important for the transport of chelants into the shoots, is efficiently blocked between shoots and seeds. PMID- 18512940 TI - High-level expression of recombinant beta-galactosidases in Lactobacillus plantarum and Lactobacillus sakei using a Sakacin P-based expression system. AB - This work presents the cloning and expression of the genes encoding heterodimeric beta-galactosidases from Lactobacillus reuteri L103, Lactobacillus acidophilus R22, Lactobacillus plantarum WCFS1, and Lactobacillus sakei Lb790. These enzymes consist of two subunits of approximately 73 and 35 kDa, which are encoded by two overlapping genes, lacL and lacM, respectively. We have cloned these genes into the lactobacillal expression vectors pSIP403 and pSIP409, which are based on the sakacin P operon of L. sakei ( Sorvig et al. Microbiology 2005, 151, 2439- 2449 ), and expressed them in the host strains L. plantarum WCFS1 and L. sakei Lb790. Results varied considerably, ranging from 2.23 to 61.1 U/mg of beta-galactosidase activity, depending on the origin of the lacLM genes, the host strain, and the expression vector used. Highest expression levels were obtained in a laboratory cultivation of L. plantarum WCFS1 harboring the plasmid pEH3R containing the lacLM gene from L. reuteri L103. These cultivations yielded approximately 23 000 U of beta-galactosidase activity per liter, corresponding to the formation of roughly 100 mg of recombinant protein per liter of fermentation medium, and beta galactosidase levels amounted to 55% of the total intracellular protein of the host organism. To further verify the suitability of this expression system, recombinant beta-galactosidase from L. reuteri was purified to apparent homogeneity. The properties of the purified enzyme were essentially identical with the properties of purified native beta-galactosidase from L. reuteri L103. The presented results lead the way to efficient overproduction of beta galactosidase in a food-grade expression system, which is of high interest for applications in food industry. PMID- 18512941 TI - Development of a real-time PCR for the detection of lupine DNA (lupinus species) in foods. AB - Lupine flour, protein, and fiber have become common ingredients in food products. The association of lupine-related allergic incidents with peanut allergy is a cause for concern as the latter may bring about severe reactions. In this study, a hybridization probe-based real-time PCR assay for the detection of lupine DNA in foods was developed. Particular attention was paid to the specificity of the method, which was verified by analysis of DNA extracts from more than 50 potential food ingredients such as legumes, cereals, seeds, nuts, spices, fruits, and meat. The limit of detection of the method was determined as 0.1 mg/kg. The successful detection of the presence/absence of lupine DNA in 20 samples proved the suitability of the assay for the analysis of frequently encountered food matrices. PMID- 18512942 TI - In vitro activity of almond skin polyphenols for scavenging free radicals and inducing quinone reductase. AB - Observational studies and clinical trials suggest nut intake, including almonds, is associated with an enhancement in antioxidant defense and a reduction in the risk of cancer and cardiovascular disease. Almond skins are rich in polyphenols (ASP) that may contribute to these putative benefits. To assess their potential mechanisms of action, we tested the in vitro effect of ASP extracted with methanol (M) or a gastrointestinal juice mimic (GI) alone or in combination with vitamins C (VC) or E (VE) (1-10 micromol/L) on scavenging free radicals and inducing quinone reductase (QR). Flavonoid profiles from ASP-M and -GI extracts were different from one another. ASP-GI was more potent in scavenging HOCl and ONOO (-) radicals than ASP-M. In contrast, ASP-M increased and ASP-GI decreased QR activity in Hepa1c1c7 cells. Adding VC or VE to ASP produced a combination- and dose-dependent action on radical scavenging and QR induction. In comparison to their independent actions, ASP-M plus VC were less potent in scavenging DPPH, HOCl, ONOO (-), and O 2 (-) (*). However, the interaction between ASP-GI plus VC promoted their radical scavenging activity. Combining ASP-M plus VC resulted in a synergistic interaction, inducing QR activity, but ASP-GI plus VC had an antagonistic effect. On the basis of their total phenolic content, the measures of total antioxidant activity of ASP-M and -GI were comparable. Thus, in vitro, ASP act as antioxidants and induce QR activity, but these actions are dependent upon their dose, method of extraction, and interaction with antioxidant vitamins. PMID- 18512943 TI - Nutritional property of endosperm starches from maize mutants: a parabolic relationship between slowly digestible starch and amylopectin fine structure. AB - The relationship between the slow digestion property of cooked maize starch and its molecular fine structure was investigated. Results of the in vitro Englyst assay showed a range of rapidly digestible starch (RDS) (70.1-98.9%), slowly digestible starch (SDS) (0.2-20.3%), and resistant starch (RS) (0.0-13.7%) among the tested maize mutant flour samples. Further analysis showed that amylose content was significantly correlated ( R = 0.763, P < 0.001) with RS amount but not with that of SDS, indicating that amylopectin is the starch molecule associated with SDS. Total starch debranching analysis revealed a parabolic relationship between SDS content and the weight ratio of amylopectin short chains (DP < 13, named SF) to long chains (DP >/= 13, named LF), which means amylopectin with a higher amount of either short chains or long chains can produce relatively high amounts of SDS. Furthermore, debranching analysis of the SDS materials from samples with the highest and lowest weight ratios of SF/LF (both had a high amount SDS) showed significantly different profiles, indicating there is not a uniform molecular structure for SDS. Thus, genetic mutants of maize samples have a good potential to provide raw starch materials of high nutritional quality. An additional finding showed that a simple and comparably high-throughput technique of Rapid Visco-Analyzer (RVA) can be used to screen genetic mutants on the basis of their RVA profiles. PMID- 18512944 TI - Automated gain control ion funnel trap for orthogonal time-of-flight mass spectrometry. AB - Time-of-flight mass spectrometry (TOF MS) is increasingly used in proteomics research. Herein, we report on the development and characterization of a TOF MS instrument with improved sensitivity equipped with an electrodynamic ion funnel trap (IFT) that employs an automated gain control (AGC) capability. The IFT-TOF MS was coupled to a reversed-phase capillary liquid chromatography (RPLC) separation and evaluated in experiments with complex proteolytic digests. When applied to a global tryptic digest of Shewanella oneidensis proteins, an order-of magnitude increase in sensitivity compared to that of the conventional continuous mode of operation was achieved due to efficient ion accumulation prior to TOF MS analysis. As a result of this sensitivity improvement and related improvement in mass measurement accuracy, the number of unique peptides identified in the AGC IFT mode was 5-fold greater than that obtained in the continuous mode. PMID- 18512945 TI - Uniform threshold intensity distribution-based quantitative multivariate imaging cytometry. AB - Implementation of quantitative analytical approaches to image-based cellular assays remains a major challenge. We disclose a tool to achieve automatic rapid quantitative cellular imaging analysis based on uniform threshold intensity distribution. An acousto-optic tunable filter-based, quantitative multivariate imaging cytometer was set up to elucidate drug-induced cell death dynamics via cell viability and apoptosis/necrosis measurements in the human myeloid leukemia cell line, HL-60. Cells were treated with various drugs (camptothecin, naringenin, sodium salicylate) at different concentrations and time intervals. The developed protocol can directly depict and quantitate targeted cellular moieties, subsequently enabling a method that is applicable to various cellular assays with special reference to next generation drug discovery screening. This may also complement certain flow-cytometric measurements in studying quantitative physiology of cellular systems. PMID- 18512946 TI - Quartz crystal microbalance with integrated surface plasmon grating coupler. AB - We have integrated a surface plasmon grating coupler into a quartz crystal microbalance (QCM) for studying surface association/dissociation reactions. In the integrated system only QCM measurement is needed to record both the optical and the acoustic signals in the same association/dissociation reaction. This integration considerably simplifies a conventional combination instrument of a grating-coupled surface plasmon resonance (SPR) spectrometer and a quartz crystal microbalance by eliminating a number of SPR components. Moreover, in the integrated system detection of the light reflections is not needed by which one bypasses the interference problem caused by two coherent light reflections off the glass window used to seal the fluid sample and off the sensor surface. The utility of the integrated system is demonstrated using a layer-by-layer polyelectrolyte multilayer deposition protocol, in which the complete features of a conventional grating-coupled SPR/QCM combination instrument are retained, including detection of optical and acoustic changes, as well as monitoring of adsorption kinetics. PMID- 18512947 TI - Mass spectrometry-based metabolomics: accelerating the characterization of discriminating signals by combining statistical correlations and ultrahigh resolution. AB - A strategy combining autocorrelation matrices and ultrahigh resolution mass spectrometry (MS) was developed to optimize the characterization of discriminating ions highlighted by metabolomics. As an example, urine samples from rats treated with phenobarbital (PB) were analyzed by ultrahigh-pressure chromatography with two different eluting conditions coupled to time-of-flight mass spectrometric detection in both the positive and negative electrospray ionization modes. Multivariate data analyses were performed to highlight discriminating variables from several thousand detected signals: a few hundred signals were found to be affected by PB, whereas a few tenths of them were linked to its metabolism. Autocorrelation matrices were then applied to eliminate adduct and fragment ions. Finally, the characterization of the ions of interest was performed with ultrahigh-resolution mass spectrometry and sequential MS(n) experiments, by using a LC-LTQ-Orbitrap system. The use of different eluting conditions was shown to drastically impact on the chromatographic retention and ionization of compounds, thus providing a way to obtain more exhaustive metabolic fingerprints, whereas autocorrelation matrices allowed one to focus the identification work on the most relevant ions. By using such an approach, 14 PB metabolites were characterized in rat urines, some of which have not been reported in the literature. PMID- 18512948 TI - Particle sorting in a mini step-split-flow thin channel: influence of hydrodynamic shear on transversal migration. AB - A mini splitterless-split-flow thin fractionation (SPLITT) device has been developed to achieve fast separations of micrometer-sized species. In this device, inlet and outlet steps have replaced the splitters, which are common to conventional SPLITT channels. By elimination of the splitters, it becomes straightforward to reduce channel dimensions while maintaining the classic method of fabrication. Reduced dimension channels allow high axial velocity at relatively low flow rate. These high axial velocities generate an enhancement of inertial lift forces and hydrodynamic shear-induced diffusion. Experiments carried out with particulate and biological species in a mini step-SPLITT channel demonstrate that these hydrodynamic effects yield highly enriched fractions of smaller species from binary mixtures. PMID- 18512949 TI - Fluorescein as a versatile tag for enhanced selectivity in analyzing cysteine containing proteins/peptides using mass spectrometry. AB - Fluorescence-based tagging in proteomics is useful in tracking and quantifying target proteins during sample preparation or chromatographic processes. In this study, we report a novel cysteinyl tagging method using a popular fluorophore, fluorescein derivative. Such visible dyes were shown to have multiple unique characteristics, including a unique reporter ion containing the dye moiety caused by collision-induced dissociation (CID) and high affinity toward multicarboxylate functional groups, which could be useful for enhanced selectivity in MS-based proteomics. We used sulfhydryl-reactive 5-iodoacetamidofluorescein to target cysteinyl residues on the intact protein of ovalbumin and bovine serum albumin as well as proteins in MCF-7 cells. After trypsin digestion, the digests were analyzed by nanoLC-ESI-Q-TOF or MALDI-TOF. The resulting MS spectra of tryptic fragments were similar to those of unlabeled or iodoacetamide-derivatized proteins, and the MS/MS fragmentation of all fluorescein-tagged peptides was readily interpretable with intact label. Thus, fluorescein-derivatized proteins can be identified by automatic mass mapping or peptide sequencing with high confidence. It is notable that, in MS/MS mode, a strong reporter ion (m/z 422) containing the fluorescein moiety was readily detected and was believed to derive from the immonium fragment of fluorescein-labeled cysteine residues, f C (m/z 463), under CID conditions. Using a precursor scan of the reporter ion, a cysteinyl protein, ovomucoid, was identified to be present in the ovalbumin sample as an impurity. The fluorescein derivatives were further shown to have high affinities toward metal-chelating materials that have iminodiacetic acid functional groups either with or without the presence of bound metal ions. When coupling with stable isotope dimethyl labeling, fluorescein-tagged peptides could be selectively enriched, identified, and quantified. In view of its popularity, visible tracking, and unique characteristics for developing selective methods, fluorescein tagging holds great promises for targeting proteomics. PMID- 18512950 TI - Mu-trap for the SALDI-MS screening of organic compounds prior to LC/MS analysis. AB - A procedure for rapidly screening and quantitatively analyzing organic molecules is presented, in which a miniaturized solid-phase extraction (SPE) cartridge containing 0.6 mg of graphitized carbon black (the GCB-mu-trap) is used for sample pretreatment. Then surface-assisted laser desorption ionization time-of flight mass spectrometry (SALDI-TOF-MS) screening is followed by liquid chromatography/mass spectrometry (LC/MS) for robust quantitative analysis of samples containing analytes of interest. Liquid samples with volumes up to 100 mL were extracted using the GCB-mu-trap, and SALDI screening was performed by transferring a few particles of the GCB 4 sorbent from the mu-trap onto a stainless steel plate. Analytes were then simply ionized and desorbed by irradiating the GCB 4 particles without any further pretreatment. GCB 4 was found to be an excellent surface for the SALDI analysis of small molecules, providing spectra with very clean backgrounds. The small size of the cartridge (micropipet filter tip) results in enrichment of the analytes on a small surface area, affording low SALDI-TOF-MS detection limits. Furthermore, the removal of just a few particles from the mu-trap does not significantly affect the subsequent quantitative determination. This approach offers considerable reductions in analytical costs by eliminating unnecessary SPE-LC/MS analyses. PMID- 18512951 TI - Top-down identification and quantification of stable isotope labeled proteins from Aspergillus flavus using online nano-flow reversed-phase liquid chromatography coupled to a LTQ-FTICR mass spectrometer. AB - Online liquid chromatography-mass spectrometric (LC-MS) analysis of intact proteins (i.e., top-down proteomics) is a growing area of research in the mass spectrometry community. A major advantage of top-down MS characterization of proteins is that the information of the intact protein is retained over the vastly more common bottom-up approach that uses protease-generated peptides to search genomic databases for protein identification. Concurrent to the emergence of top-down MS characterization of proteins has been the development and implementation of the stable isotope labeling of amino acids in cell culture (SILAC) method for relative quantification of proteins by LC-MS. Herein we describe the qualitative and quantitative top-down characterization of proteins derived from SILAC-labeled Aspergillus flavus using nanoflow reversed-phase liquid chromatography directly coupled to a linear ion trap Fourier transform ion cyclotron resonance mass spectrometer (nLC-LTQ-FTICR-MS). A. flavus is a toxic filamentous fungus that significantly impacts the agricultural economy and human health. SILAC labeling improved the confidence of protein identification, and we observed 1318 unique protein masses corresponding to 659 SILAC pairs, of which 22 were confidently identified. However, we have observed some limiting issues with regard to protein quantification using top-down MS/MS analyses of SILAC-labeled proteins. The role of SILAC labeling in the presence of competing endogenously produced amino acid residues and its impact on quantification of intact species are discussed in detail. PMID- 18512952 TI - The electronic structure of the Cys-Tyr(*) free radical in galactose oxidase determined by EPR spectroscopy. AB - Galactose oxidase is a metalloenzyme containing a novel metalloradical complex in its active site, comprised of a mononuclear copper ion associated with a protein free radical. The free radical has been shown to be localized on an intrinsic redox cofactor, 3'-(S-cysteinyl)tyrosine (Cys-Tyr), formed by a posttranslational covalent coupling of tyrosine and cysteine side chains in a self-processing reaction. The role of the thioether linkage in the function of the Cys-Tyr cofactor is unresolved, and some computational studies have suggested that the thioether substituent has a negligible effect on the properties of the tyrosyl free radical. In order to address this question experimentally, we have incorporated site-selectively labeled tyrosine ((2)H, (13)C, (17)O) into galactose oxidase using an engineered tyrosine auxotroph strain of Pichia pastoris . (33)S was also incorporated into the protein. EPR spectra for the Cys Tyr(*) free radical in each of these isotopic variants were analyzed to extract nuclear hyperfine parameters for comparison with theoretical predictions, and the unpaired spin distribution in the free radical was reconstructed from the hyperfine data. These labeling studies allow the first comprehensive experimental evaluation of the effect of the thioether linkage on the properties of Cys-Tyr(*) and indicate that previous calculations significantly underestimated the contribution of this feature to the electronic ground state of the free radical. PMID- 18512953 TI - Dynamics of Rhodobacter capsulatus [2FE-2S] ferredoxin VI and Aquifex aeolicus ferredoxin 5 via nuclear resonance vibrational spectroscopy (NRVS) and resonance Raman spectroscopy. AB - We have used (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to study the Fe(2)S(2)(Cys)(4) sites in oxidized and reduced [2Fe-2S] ferredoxins from Rhodobacter capsulatus (Rc FdVI) and Aquifex aeolicus (Aa Fd5). In the oxidized forms, nearly identical NRVS patterns are observed, with strong bands from Fe-S stretching modes peaking around 335 cm(-1), and additional features observed as high as the B(2u) mode at approximately 421 cm(-1). Both forms of Rc FdVI have also been investigated by resonance Raman (RR) spectroscopy. There is good correspondence between NRVS and Raman frequencies, but because of different selection rules, intensities vary dramatically between the two kinds of spectra. For example, the B(3u) mode at approximately 288 cm(-1), attributed to an asymmetric combination of the two FeS(4) breathing modes, is often the strongest resonance Raman feature. In contrast, it is nearly invisible in the NRVS, as there is almost no Fe motion in such FeS(4) breathing. NRVS and RR analysis of isotope shifts with (36)S-substituted into bridging S(2-) ions in Rc FdVI allowed quantitation of S(2-) motion in different normal modes. We observed the symmetric Fe-Fe stretching mode at approximately 190 cm(-1) in both NRVS and RR spectra. At still lower energies, the NRVS presents a complex envelope of bending, torsion, and protein modes, with a maximum at 78 cm(-1). The (57)Fe partial vibrational densities of states (PVDOS) were interpreted by normal-mode analysis with optimization of Urey-Bradley force fields. Progressively more complex D(2h) Fe(2)S(2)S'(4), C(2h) Fe(2)S(2)(SCC)(4), and C(1) Fe(2)S(2)(Cys)(4) models were optimized by comparison with the experimental spectra. After modification of the CHARMM22 all-atom force field by the addition of refined Fe-S force constants, a simulation employing the complete protein structure was used to reproduce the PVDOS, with better results in the low frequency protein mode region. This process was then repeated for analysis of data on the reduced FdVI. Finally, the degree of collectivity was used to quantitate the delocalization of the dynamic properties of the redox-active Fe site. The NRVS technique demonstrates great promise for the observation and quantitative interpretation of the dynamical properties of Fe-S proteins. PMID- 18512954 TI - Characterization of chondroitin sulfate lyase ABC from Bacteroides thetaiotaomicron WAL2926. AB - Chondroitin sulfate ABC lyase (ChonABC) is an enzyme with broad specificity that depolymerizes via beta-elimination chondroitin sulfate (CS) and dermatan sulfate (DS) glycosaminoglycans (GAGs). ChonABC eliminates the glycosidic bond of its GAG substrates on the nonreducing end of their uronic acid component. This lyase possesses the unusual ability to act on both epimers of uronic acid, either glucuronic acid present in CS or iduronic acid in DS. Recently, we cloned, purified, and determined the three-dimensional structure of a broad specificity chondroitin sulfate ABC lyase from Bacteroides thetaiotaomicron (BactnABC) and identified two sets of catalytic residues. Here, we report the detailed biochemical characterization of BactnABC together with extensive site-directed mutagenesis resulting in characterization of the previously identified active site residues. BactnABC's catalysis is stimulated by Ca(2+) and Mg(2+) cations, particularly against DS. It displays extremely low activity toward hyaluronic acid and no activity toward heparin/heparan sulfate. Degradation of CS and DS by BactnABC yields only disaccharide products, pointing to an exolytic mode of action. The kinetic evaluations of the active-site mutants indicate that CS and DS substrates bind in the same active site, which is accompanied by a conformational change bringing the two sets of active site residues together. Conservative replacements of key residues suggest that His345 plays the role of a general base, initiating the degradation by abstracting the C5 bound proton from DS substrates, whereas either Tyr461 or His454 perform the equivalent role for CS substrates. Tyr461 is proposed, as well, to serve as general acid, completing the degradation of both CS and DS by protonating the leaving group. PMID- 18512955 TI - Engineering the glucansucrase GTFR enzyme reaction and glycosidic bond specificity: toward tailor-made polymer and oligosaccharide products. AB - Two long-standing questions about glucansucrases (EC 2.4.1.5) are how they control oligosaccharide versus polysaccharide synthesis and how they direct their glycosidic linkage specificity. This information is required for the production of tailor-made saccharides. Mutagenesis promises to be an effective tool for enzyme engineering approaches for altering the regioselectivity and acceptor substrate specificity. Therefore, we chose the most conserved motif around the transition state stabilizer in glucansucrases for a random mutagenesis of the glucansucrase GTFR of Streptococcus oralis, yielding different variants with altered reaction specificity. Modifications at position S628 achieved by saturation mutagenesis guided the reaction toward the synthesis of short chain oligosaccharides with a drastically increased yield of isomaltose (47%) or leucrose (64%). Alternatively, GTFR variant R624G/V630I/D717A exhibited a drastic switch in regioselectivity from a dextran type with mainly alpha-1,6-glucosidic linkages to a mutan type polymer with predominantly alpha-1,3-glucosidic linkages. Targeted modifications demonstrated that both mutations near the transition state stabilizer, R624G and V630I, are contributing to this alteration. It is thus shown that mutagenesis can guide the transglycosylation reaction of glucansucrase enzymes toward the synthesis of (a) various short chain oligosaccharides or (b) novel polymers with completely altered linkages, without compromising their high transglycosylation activity and efficiency. PMID- 18512956 TI - Measuring the interaction of urea and protein-stabilizing osmolytes with the nonpolar surface of hydroxypropylcellulose. AB - The interaction of urea and several naturally occurring protein-stabilizing osmolytes, glycerol, sorbitol, glycine betaine, trimethylamine oxide (TMAO), and proline, with condensed arrays of a hydrophobically modified polysaccharide, hydroxypropylcellulose (HPC), has been inferred from the effect of these solutes on the forces acting between HPC polymers. Urea interacts only very weakly. The protein-stabilizing osmolytes are strongly excluded. The observed energies indicate that the exclusion of the protein-stabilizing osmolytes from protein hydrophobic side chains would add significantly to protein stability. The temperature dependence of exclusion indicates a significant contribution of enthalpy to the interaction energy in contrast to expectations from "molecular crowding" theories based on steric repulsion. The dependence of exclusion on the distance between HPC polymers rather indicates that perturbations of water structuring or hydration forces underlie exclusion. PMID- 18512957 TI - Crystal structures of substrate-free and retinoic acid-bound cyanobacterial cytochrome P450 CYP120A1. AB - The crystal structures of substrate-free and all-trans-retinoic acid-bound CYP120A1 from Synechocystis sp. PCC 6803 were determined at 2.4 and 2.1 A resolution, respectively, representing the first structural characterization of a cyanobacterial P450. Features of CYP120A1 not observed in other P450 structures include an aromatic ladder flanking the channel leading to the active site and a triple-glycine motif within SRS5. Using spectroscopic methods, CYP120A1 is shown to bind 13-cis-retinoic acid, 9-cis-retinoic acid, and retinal with high affinity and dissociation constants of less than 1 microM. Metabolism of retinoic acid by CYP120A1 suggests that CYP120A1 hydroxylates a variety of retinoid derivatives in vivo. On the basis of the retinoic acid-bound CYP120A1 crystal structure, we propose that either carbon 2 or the methyl groups (C16 or C17) of the beta-ionone ring are modified by CYP120A1. PMID- 18512958 TI - Miscoding properties of 6alpha- and 6beta-diastereoisomers of the N(2)-(estradiol 6-yl)-2'-deoxyguanosine DNA adduct by Y-family human DNA polymerases. AB - Treatment with estrogen increases the risk of breast, ovary, and endometrial cancers in women. DNA damage induced by estrogen is thought to be involved in estrogen carcinogenesis. In fact, Y-family human DNA polymerases (pol) eta and kappa, which are highly expressed in the reproductive organs, miscode model estrogen-derived DNA adducts during DNA synthesis. Since the estrogen-DNA adducts are a mixture of 6alpha- and 6beta-diastereoisomers of dG-N(2)-6-estrogen or dA N(6)-6-estrogen, the stereochemistry of each isomeric adduct on translesion synthesis catalyzed by DNA pols has not been investigated. We have recently established a phosphoramidite chemical procedure to insert 6alpha- or 6beta isomeric N(2)-(estradiol-6-yl)-2'-deoxyguanosine (dG-N(2)-6-E(2)) into oligodeoxynucleotides. Using such site-specific modified oligomer as a template, the specificity and frequency of miscoding by dG-N(2)-6alpha-E(2) or dG-N(2) 6beta-E(2) were explored using pol eta and a truncated form of pol kappa (pol kappaDeltaC). Translesion synthesis catalyzed by pol eta bypassed both the 6alpha and 6beta-isomers of dG-N(2)-6-E(2), with a weak blockage at the adduct site, while translesion synthesis catalyzed by pol kappaDeltaC readily bypassed both isomeric adducts. Quantitative analysis of base substitutions and deletions occurring at the adduct site showed that pol kappaDeltaC was more efficient than pol eta by incorporating dCMP opposite both 6alpha- and 6beta-isomeric dG-N(2)-6 E(2) adducts. The miscoding events occurred more frequently with pol eta, but not with pol kappaDeltaC. Pol eta promoted incorporation of dAMP and dTMP at both the 6alpha- and 6beta-isomeric adducts, generating G --> T transversions and G --> A transitions. One- and two-base deletions were also formed. The 6alpha-isomeric adduct promoted slightly lower frequency of dCMP incorporation and higher frequency of dTMP incorporation and one-base deletions, compared with the 6beta isomeric adduct. These observations were supported by steady-state kinetic studies. Taken together, the miscoding property of the 6alpha-isomeric dG-N(2)-6 E(2) is likely to be similar to that of the 6beta-isomeric adduct. PMID- 18512959 TI - The thymine-DNA glycosylase regulatory domain: residual structure and DNA binding. AB - Thymine-DNA glycosylases (TDGs) initiate base excision repair by debasification of the erroneous thymine or uracil nucleotide in G.T and G.U mispairs which arise at high frequency through spontaneous or enzymatic deamination of methylcytosine and cytosine, respectively. Human TDG has furthermore been shown to have a functional role in transcription and epigenetic regulation through the interaction with transcription factors from the nuclear receptor superfamily, transcriptional coregulators, and a DNA methyltransferase. The TDG N-terminus encodes regulatory functions, as it assures both G.T versus G.U specificity and contains the sites for interaction and posttranslational modification by transcription-related activities. While the molecular function of the evolutionarily conserved central catalytic domain of TDG in base excision repair has been elucidated by determination of its three-dimensional structure, the mechanisms by which the N-terminus exerts its regulatory roles, as well as the function of TDG in transcription regulation, remain to be understood. We describe here the residual structure of the TDG N-terminus in both contexts of the isolated domain and the entire protein. These studies lead to the characterization of a small structural domain in the TDG N-terminal region preceding the catalytic core and coinciding with the region of functional regulation of TDG's activities. This regulatory domain exhibits a small degree of organization and is implicated in dynamic molecular interactions with the catalytic domain and nonselective interactions with double-stranded DNA, providing a molecular explanation for the evolutionarily acquired G.T mismatch processing activity of TDG. PMID- 18512960 TI - Product specificity and mechanism of protein lysine methyltransferases: insights from the histone lysine methyltransferase SET8. AB - Molecular dynamics simulations employing a molecular mechanics (MM) force field and hybrid quantum mechanics (QM) and MM (QM/MM) have been carried out to investigate the product specificity and mechanism of the histone H4 lysine 20 (H4 K20) methylation by human histone lysine methyltransferase SET8. At neutral pH, the target lysine is available to only the enzyme in the protonated state. The first step in the methylation reaction must be deprotonation of the lysine target which is followed by the (+)AdoMet methylation of the neutral lysine [Enz.Lys CH(2)-NH(3)(+).(+)AdoMet --> H(+) + Enz.Lys-CH(2)-NH(2).(+)AdoMet -->--> Enz.Lys CH(2)-N(Me)H(2)(+).AdoHcy]. The electrostatic interactions between two positive charges on (+)AdoMet and Lys20-NH(3)(+) decrease the pK(a) of Lys20-NH(3)(+). Upon formation of Enz.Lys-NH(3)(+).(+)AdoMet, a water channel by which the proton escapes to the outer solvent phase is formed. The formation of a water channel for the escape of a proton from Lys20-N(Me)H(2)(+) in Enz.Lys20 N(Me)H(2)(+).(+)AdoMet is not formed because the methyl substituent blocks the starting of the water channel. Thus, a second methylation does not take place. The dependence of the occurrence of methyl transfer on the formation of a water channel in SET8 is in accord with our previous reports on product specificity by histone lysine monomethyltransferase SET7/9, large subunit lysine dimethyltransferase (LSMT), and viral histone lysine trimethyltransferase (vSET). The average value of the experimental DeltaG(E)() for the six lysine methyl transfer reactions catalyzed by vSET, LSMT, and SET7/9 with p53 as a substrate is 22.1 +/- 1.0 kcal/mol, and the computed average (DeltaG(C)()) is 22.2 +/- 0.8 kcal/mol. In this study, the computed free energy barrier of the methyl transfer reaction [Lys20-NH(2) + (+)AdoMet --> Lys20-N(Me)H(2)(+) + AdoHcy] catalyzed by SET8 is 20.8 kcal/mol. This is in agreement with the value of 20.6 kcal/mol calculated from the experimental rate constant (0.43 +/- 0.02 min(-1)). Our bond order computations establish that the H4-K20 monomethylation in SET8 is a concerted linear S(N)2 displacement reaction. PMID- 18512961 TI - Orientation of the Escherichia coli outer membrane protein OmpX in phospholipid bilayer membranes determined by solid-State NMR. AB - The solid-state NMR orientation-dependent frequencies measured for membrane proteins in macroscopically oriented lipid bilayers provide precise orientation restraints for structure determination in membranes. Here we show that this information can also be used to supplement crystallographic structural data to establish the orientation of a membrane protein in the membrane. This is achieved by incorporating a few orientation restraints, measured for the Escherichia coli outer membrane protein OmpX in magnetically oriented lipid bilayers (bicelles), in a simulated annealing calculation with the coordinates of the OmpX crystal structure. The (1)H-(15)N dipolar couplings measured for the seven Phe residues of OmpX in oriented bilayers can be assigned by back-calculation of the NMR spectrum from the crystal structure and are sufficient to establish the three dimensional orientation of the protein in the membrane, while the (15)N chemical shifts provide a measure of cross-validation for the analysis. In C14 lipid bilayers, OmpX adopts a transmembrane orientation with a 7 degrees tilt of its beta-barrel axis relative to the membrane normal, matching the hydrophobic thickness of the barrel with that of the membrane. PMID- 18512962 TI - Structural comparison of crystal and solution states of the 138 kDa complex of methylamine dehydrogenase and amicyanin from Paracoccus versutus. AB - Methylamine can be used as the sole carbon source of certain methylotrophic bacteria. Methylamine dehydrogenase catalyzes the conversion of methylamine into formaldehyde and donates electrons to the electron transfer protein amicyanin. The crystal structure of the complex of methylamine dehydrogenase and amicyanin from Paracoccus versutus has been determined, and the rate of electron transfer from the tryptophan tryptophylquinone cofactor of methylamine dehydrogenase to the copper ion of amicyanin in solution has been determined. In the presence of monovalent ions, the rate of electron transfer from the methylamine-reduced TTQ is much higher than in their absence. In general, the kinetics are similar to those observed for the system from Paracoccus denitrificans. The complex in solution has been studied using nuclear magnetic resonance. Signals of perdeuterated, (15)N-enriched amicyanin bound to methylamine dehydrogenase are observed. Chemical shift perturbation analysis indicates that the dissociation rate constant is approximately 250 s(-1) and that amicyanin assumes a well defined position in the complex in solution. The most affected residues are in the interface observed in the crystal structure, whereas smaller chemical shift changes extend to deep inside the protein. These perturbations can be correlated to small differences in the hydrogen bond network observed in the crystal structures of free and bound amicyanin. This study indicates that chemical shift changes can be used as reliable indicators of subtle structural changes even in a complex larger than 100 kDa. PMID- 18512963 TI - Functional and structural characterization of DR_0079 from Deinococcus radiodurans, a novel Nudix hydrolase with a preference for cytosine (deoxy)ribonucleoside 5'-Di- and triphosphates. AB - The genome of the extremely radiation resistant bacterium Deinococcus radiodurans encodes 21 Nudix hydrolases, of which only two have been characterized in detail. Here we report the activity and crystal structure for DR_0079, the first Nudix hydrolase observed to have a marked preference for cytosine ribonucleoside 5' diphosphate (CDP) and cytosine ribonucleoside 5'-triphosphate (CTP). After CDP and CTP, the next most preferred substrates for DR_0079, with a relative activity of <50%, were the corresponding deoxyribose nucleotides, dCDP and dCTP. Hydrolase activity at the site of the phosphodiester bond was corroborated using (31)P NMR spectroscopy to follow the phosphorus resonances for three substrates, CDP, IDP, and CTP, and their hydrolysis products, CMP + P(i), IMP + P(i), and CMP + PP(i), respectively. Nucleophilic substitution at the beta-phosphorus of CDP and CTP was established, using (31)P NMR spectroscopy, by the appearance of an upfield shifted P(i) resonance and line-broadened PP(i) resonance, respectively, when the hydrolysis was performed in 40% H(2)(18)O-enriched water. The optimal activity for CDP was at pH 9.0-9.5 with the reaction requiring divalent metal cation (Mg(2+) > Mn(2+) > Co(2+)). The biochemical data are discussed with reference to the crystal structure for DR_0079 that was determined in the metal-free form at 1.9 A resolution. The protein contains nine beta-strands, three alpha-helices, and two 3(10)-helices organized into three subdomains: an N-terminal beta-sheet, a central Nudix core, and a C-terminal helix-turn-helix motif. As observed for all known structures of Nudix hydrolases, the alpha-helix of the "Nudix box" is one of two helices that sandwich a "four-strand" mixed beta-sheet. To identify residues potentially involved in metal and substrate binding, NMR chemical shift mapping experiments were performed on (15)N-labeled DR_0079 with the paramagnetic divalent cation Co(2+) and the nonhydrolyzable substrate thymidine 5'-O (alpha,beta-methylenediphosphate) and the results mapped onto the crystal structure. PMID- 18512964 TI - Computational modeling of substituent effects on phenol toxicity. AB - Standard computational models of cytotoxicity of substituted phenols relate the toxicity to a set of quatitative structure-activity relationship (QSAR) descriptors such as log P, p K a, OH bond dissociation enthalpy (BDE), etc. Implicit in this approach is the idea that the phenoxyl radical is disruptive to the cell and factors increasing its production rate will enhance the toxicity. To improve the QSAR correlations, substituents are usually divided into electron donating groups (EDG) and electron-withdrawing groups (EWG), which are treated separately and thought to follow different mechanisms of toxicity. In this paper, we focus on one important aspect of toxicity, the rate constant for production of phenoxyl radical. Activation energies are obtained for the reaction of X-phenol with peroxyl radical by using the Evans-Polanyi principle, giving rate constants as a function of DeltaBDE values for both EDG and EWG sets. We show that (i) a plot of log k for phenoxyl formation vs DeltaBDE shows a double set of straight lines with different slopes, justifying the usual EDG and EWG separation but without requiring any change in mechanism; (ii) the same method can be effectively used for different target radicals (e.g., tert-butoxyl) or different sets of parent compounds (e.g., substituted catechols), thus giving a useful general approach to analysis of toxicity data; (iii) regions of constant toxicity in all cases are predicted; and (iv) we argue that competing parallel mechanisms of toxicity are likely to be dominant for EWG-substituted phenols. PMID- 18512965 TI - Activation of Nrf2 in defense against cadmium-induced oxidative stress. AB - Exposure to cadmium (Cd) elicits a range of adverse responses including oxidative damage and cancer. The molecular targets of Cd remain largely unidentified. Here, we analyzed the function and signal transduction of transcription factor Nrf2 in protection against Cd-induced oxidative stress. Wild-type (Nrf2 (+/+)) mouse embryonic fibroblasts (MEF) produced reactive oxygen species (ROS) at a low level, whereas treatment with Cd significantly increased the ROS production. On the other hand, Nrf2 knockout (Nrf2 (-/-)) MEF cells exhibited an elevated level of ROS under a basal condition, and Cd dramatically increased the ROS production at concentrations as low as 2 microM, resulting in increased sensitivity to Cd induced cell death. Cd induced the basal and inducible expression of cytoprotective enzymes NQO1 and HO1 in WT MEF cells, but induction was lost in Nrf2 (-/-) MEF cells. Induction of the genes required antioxidant response elements (ARE) as Cd drove ARE-dependent reporter expression and Cd-activated Nrf2 bound to endogenous AREs in mouse hepa1c1c7 cells. Activation of Nrf2 by Cd involved stabilization of the Nrf2 protein, increased formation of Nrf2/Keap1 complex in the cytoplasm, translocation of the complex into the nucleus, and subsequently disruption of the complex. Lastly, Nrf2 was found ubiquitinated in the cytoplasm but deubiquitinated in the nucleus. The study provided a mechanistic transcriptional model in which Cd activates Nrf2 through a metal activated signaling pathway involving a dynamic interplay between ubiquitination/deubiquitination and complex formation/dissociation of Nrf2 and Keap1. PMID- 18512966 TI - Induction of apoptosis by DC-81-indole conjugate agent through NF-kappaB and JNK/AP-1 pathway. AB - DC-81, an antitumor antibiotic produced by Streptomyces species, belongs to the pyrrolo[2,1- c][1,4]benzodiazepine (PBD) family, which are potent inhibitors of nucleic acid synthesis. We previously reported an efficient synthesis of PBD hybrids linked with indole carboxylates. Recently, we have also shown that a PBD hybrid (IN6CPBD) agent can activate the apoptotic pathway mediated by mitochondria. In this study, we will examine the transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) that functionally regulate cell proliferation, transformation, and apoptosis. To investigate the IN6CPBD-induced alterations in NF-kappaB and AP-1 activity that involve cell cycle regulation, we exposed human melanoma A375 cells to different concentrations of IN6CPBD. Our data revealed that treatment of A375 cells with IN6CPBD resulted in a marked loss of cells from the G2/M phase of the cell cycle and an increase in Ca (2+) and cAMP and promoted phosphorylation of Jun N terminal kinase (JNK) expression. By using the luciferase reporter assay, the NF kappaB activities were decreased; however, AP-1 activity was further enhanced after A375 cells were treated with graded concentrations of IN6CPBD. Blockade of NF-kappaB or JNK activity further enhanced caspase-3 substrate PARP cleavage and subsequent apoptotic cell death. PMID- 18512967 TI - Protein N-acylation: H2O2-mediated covalent modification of protein by lipid peroxidation-derived saturated aldehydes. AB - Various lines of evidence indicate that the oxidative modification of protein and the subsequent accumulation of the degenerated proteins have been found in cells and tissues during aging, oxidative stress, and in a variety of pathological states. The critical agents that give rise to this protein degeneration may be represented by aldehydes. Although the covalent modification of proteins by aldehydes alone has been well-studied, the effect of reactive oxygen species, such as H2O2, upon aldehyde modification of the protein has received little attention. We have now established a unique protein modification in which H2O2 and, to a lesser extent, alkyl hydroperoxides mediate the binding of alkanals to the lysine residues of protein to generate structurally unusual N-acylation products. Upon the reaction of a lysine-containing peptide, N(alpha) benzoylglycyl-lysine, with hexanal in the presence of H2O2, a product containing one molecule of hexanal per peptide was detected. On the basis of the chemical and spectroscopic evidence, the product was identified to be the acylation product, N(epsilon)-hexanoyllysine. H2O2 mediated the N-acylation of the lysine derivative by the saturated aldehydes of 1-6 carbons in length. The H2O2-mediated acylation of the protein was immunochemically confirmed by reaction of the proteins with hexanal in the presence of H2O2. Furthermore, the enhanced N acylations (N-acetylation and N-hexanoylation) were also observed in the kidney of rats exposed to ferric nitrilotriacetate, a well-characterized inducer of oxidative stress. Mechanistic studies using a phosphonium lysine derivative suggest a Baeyer-Villiger-like reaction proceeding through peroxide addition to the aldehyde Schiff base. These data suggest that the hydroperoxides, including H2O2, might be involved not only in the oxidative modification of protein but also in the covalent binding of the saturated aldehydes to proteins under oxidative stress. PMID- 18512968 TI - Galloylated catechins as potent inhibitors of hypochlorous acid-induced DNA damage. AB - Hypochlorous acid (HOCl), a strong oxidant derived from myeloperoxidase in neutrophils and macrophages, can chlorinate DNA bases at the site of inflammation. Because little is known about the protective role of natural antioxidants, such as polyphenols, for the myeloperoxidase-derived DNA damage, we screened the inhibitory effects of various phenolic antioxidants on the chlorination of the 2'-deoxycytidine residue by HOCl in vitro and found that green tea catechins, especially (-)-epicatechin gallate (ECg) and (-) epigallocatechin gallate (EGCg), significantly inhibited the chlorination. These catechins also reduced nucleoside- and taurine-chloramines, which can induce secondary oxidative damage, into their native forms. Mass spectrometric and nuclear magnetic resonance analyses showed that ECg and EGCg can effectively scavenge HOCl and/or chloramine species resulting in the formation of mono- and dichlorinated ECg and EGCg. Using the HL-60 human leukemia cell line, it was found that ECg could efficiently accumulate in the cells. Immunocytometric analyses using antihalogenated 2'-deoxycytidine antibody showed that pretreatment of cells with ECg inhibited the HOCl-induced immunofluorescence. In addition, the chlorinated ECg derivatives were detected in the HOCl-treated HL-60 cells. These results showed that green tea catechins, especially 3-galloylated catechins, may be the plausible candidate for the prevention of inflammation-derived DNA damage and perhaps carcinogenesis. PMID- 18512970 TI - A universal approach to solvation modeling. AB - Continuum mean-field models that have been carefully designed to address the various electrostatic and nonelectrostatic interactions that develop between a molecule and a surrounding medium are particularly efficient tools for studying the effects of condensed phases on molecular structure, energetics, properties, spectra, interaction potentials, and dynamics. The SM8 model may be combined with density functional theory or Hartree-Fock theory to describe a solute's electronic structure and its self-consistent-field polarization by a solvent. A key feature is the use of class IV charge models to obtain accurate charge distributions (either in the vapor phase or in solution), even when using small basis sets that are affordable for large systems. A second key feature is that nonelectrostatic effects due to cavity formation, dispersion interactions, and changes in solvent structure are included in terms of empirical atomic surface tensions that depend on geometry but do not require atom-type assignments by the user. Use of an analytic surface area algorithm provides very stable energy gradients that allow geometry optimization in solution. The SM8 continuum model, the culmination of a series of SMx models (x = 1-8), permits the modeling of such diverse media as aqueous and organic solvents, soils, lipid bilayers, and air water interfaces. In addition to predicting accurate transfer free energies between gaseous and condensed phases or between two different condensed phases, SMx models have been useful for predicting the significant influence of condensed phases on processes associated with a change in molecular charge, including acid/base equilibria and oxidation/reduction processes. In this Account, we provide an overview of the algorithms associated with the computation of free energies of solvation in the SM8 model. We also compare the accuracies of the SM8 model with those of other continuum solvation models. Finally, we highlight applications of the SM8 models to compute ionic solvation free energies, oxidation and reduction potentials, and pK(a) values. PMID- 18512969 TI - Efficient synthesis, liquid chromatography purification, and tandem mass spectrometric characterization of estrogen-modified DNA bases. AB - Estrogens are metabolized to active quinones that modify DNA and may lead to various cancers. To extend the analytical methodology for estrogen-modified purine bases, we report here a simple modification to existing synthetic procedures that use 2-iodoxybenzoic acid (IBX) as the oxidizing agent for the reference material and putative biomarker, 4-hydroxyestrone-1-N3adenine (4-OH-E1 1-N3Ade). The reaction leads to two catechol estrogen quinones, CE1-2,3-Q and CE1 3,4-Q, both of which react via Michael additions to afford 4-OH-E1-1-N3Ade and other DNA adducts. Liquid chromatography separation permits the isolation of high purity 4-OH-E1-1-N3Ade. With this method, we also prepared single 13C and uniformly 15N (U-15N) labeled 4-OH-E1-1-N3Ade with 8-13C-labeled Ade and U-15N labeled adenosine 5'-monophosphate (AMP). The approach is also effective for the synthesis of 4-hydroxyestradiol-1-N3adenine, 4-OH-E2-1-N3Ade, and 4 hydroxyestrone(estradiol)-1-N7guanine, 4-OH-E1(E2)-1-N7Gua. The tandem mass spectra (MS2 and MS3) of 4-OH-E1-(unlabeled, 8-13C-, and U-15N-labeled)1-N3Ade and accurate mass measurements for MS2 product ions allow us to assign unambiguously the formula of fragments and delineate the fragmentation pathways. One important reaction is dehydration, which occurs at the ketone oxygen in the C 17 position of estrone. Another is loss of NH3, an ubiquitous process for purines and modified purines, which is affected by the steroid modification. Evidence from MS/MS supports the migration of H-atom(s) from estrone in the loss of NH3. An interesting interaction occurs between the steroid and the Ade in the modified base, promoting loss of CH2NH, a loss that distinguishes modified Ade from unmodified Ade. The synthesis of a stable isotope-labeled 4-OH-E1-1-N3Ade and the understanding of the fragmentation processes will enable studies aimed at the etection of naturally occurring 4-OH-E1-1-N3Ade in biological samples. PMID- 18512972 TI - Aptamer-targeted gold nanoparticles as molecular-specific contrast agents for reflectance imaging. AB - Targeted metallic nanoparticles have shown potential as a platform for development of molecular-specific contrast agents. Aptamers have recently been demonstrated as ideal candidates for molecular targeting applications. In this study, we investigated the development of aptamer-based gold nanoparticles as contrast agents, using aptamers as targeting agents and gold nanoparticles as imaging agents. We devised a novel conjugation approach using an extended aptamer design where the extension is complementary to an oligonucleotide sequence attached to the surface of the gold nanoparticles. The chemical and optical properties of the aptamer-gold conjugates were characterized using size measurements and oligonucleotide quantitation assays. We demonstrate this conjugation approach to create a contrast agent designed for detection of prostate-specific membrane antigen (PSMA), obtaining reflectance images of PSMA(+) and PSMA(-) cell lines treated with the anti-PSMA aptamer-gold conjugates. This design strategy can easily be modified to incorporate multifunctional agents as part of a multimodal platform for reflectance imaging applications. PMID- 18512973 TI - Increased throughput and reduced carryover of mass spectrometry-based proteomics using a high-efficiency nonsplit nanoflow parallel dual-column capillary HPLC system. AB - We report a new design of a fully automated, high-efficiency parallel nonsplit nanoflow capillary HPLC system, coupled on-line with linear ion trap (LTQ) and high performance nanoelectrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (nanoESI LTQ-FTICR MS). The system, intended for high throughput proteome analysis of complex protein mixtures, notably serum and plasma, consists of two reversed-phase trap columns for large volume sample injection with high speed sample loading and desalting and two reversed-phase analytical capillary columns. Through a nanoscale two-position, 10-port switching valve, the whole system is terminated by a 10 microm i.d. of nanoemitter mounted on the nanoelectrospray source in front of the sampling cone of the LTQ-FTICR MS. Gradient elution to both nanoflow-rate capillary columns is simultaneously delivered by a single HPLC system via two independent binary gradient pump systems. The parallel capillary column approach eliminates the time delays for column regeneration/equilibration since one capillary column is used for separating the sample mixtures and delivering the separated fractions to the MS, while the other capillary column is being regenerated and equilibrated. The reproducibility of retention time and peak intensity of the present automated parallel nanoflow-rate capillary HPLC system is comparable to that obtained using a single column configuration. Replicate injections of tryptic digests indicated that this system provided good reproducibility of retention time and peak area on both columns with average CV values of less than 1.08% and 7.04%, respectively. Throughput was increased to 100% for 2-h LC-MS analysis compared to the single capillary column LC-MS pipeline. Application of this system is demonstrated in a plasma proteomic study. A total of 312 868 MSMS events were acquired and 1564 proteins identified with high confidence (Protein Prophet > or = 0.9, and peptides matched > or = 2). Comparison of a series of plasma fractions run using the single-column LC-MS versus the parallel-column LC-MS demonstrated that parallel-column LC-MS system significantly reduced the sample carryover, improved MS data quality and increased the number of MS/MS sequence scan events. PMID- 18512974 TI - 2D-IR spectroscopy of the sulfhydryl band of cysteines in the hydrophobic core of proteins. AB - We investigate the sulfhydryl band of cysteines as a new chromophore for two dimensional IR (2D-IR) studies of the structure and dynamics of proteins. Cysteines can be put at almost any position in a protein by standard methods of site-directed mutagenesis and, hence, have the potential to be an extremely versatile local probe. Although being a very weak absorber in aqueous environment, the sulfhydryl group gets strongly polarized when situated in an alpha-helix inside the hydrophobic core of a protein because of a strong hydrogen bond to the backbone carbonyl group. The extinction coefficient (epsilon=150 M( 1) cm(-1)) then is sufficiently high to perform detailed 2D-IR studies even at low millimolar concentrations. Using porcine (carbonmonoxy)hemoglobin as an example, which contains two such cysteines in its wild-type form, we demonstrate that spectral diffusion deduced from the 2D-IR line shapes reports on the overall breathing of the corresponding alpha-helix. The vibrational lifetime of the sulfhydryl group (T1 approximately 6 ps) is considerably longer than that of the much more commonly used amide I mode (approximately 1.0 ps), thereby significantly extending the time window in which spectral diffusion processes can be observed. The experiments are accompanied by molecular dynamics simulations revealing a good overall agreement. PMID- 18512975 TI - Spermine-induced hybridization and charge inversion at the diffuse layer of a DNA FET. AB - Detection of DNA hybridization events by using field-effect transducers is limited by the electrolyte content of the medium. So far, DNA was thought to hybridize only in solutions with concentrated electrolytes. In these media, the interface between the transducer gate and the solution is reduced to a thin layer in close contact with the surface, and DNA is poorly detected. In the present work, this limitation is overcome by using spermine as screening polycation. Hybridization assays with polycation concentrations as low as 10 microM are reported. This ensures that hybridization takes place at the diffuse layer of the interface. The reported results suggest a charge-inversion mechanism induced by spermine. A target sequence is real-time label-free detected in the range from 10 to 500 nM. PMID- 18512976 TI - Photoresponsive J-aggregation behavior of a novel azobenzene-phthalocyanine dyad and its third-order optical nonlinearity. AB - The photoresponsive J-aggregation behaviors of a novel azobenzene-substituted zinc phthalocyanine (azo-ZnPc dyad) were studied by UV/vis, fluorescence, and (1)H NMR spectroscopy. Upon illumination with 365 nm UV light, the trans-cis isomerization of azobenzene can efficiently reduce the steric hindrance around the peripheral oxygen atom of azo-ZnPc, shortening the possible distance between two phthalocyanine molecules and, consequently, greatly improving the tendency of J-aggregation of azo-ZnPc dyad. The third-order optical nonlinearities of the photoresponsive J-aggregates (before and after illumination) were measured by a Z scan technique at 532 nm with a pulse duration of 25 ps. The Z-scan spectra revealed that all the samples possessed large positive nonlinear refraction and positive nonlinear absorption, exhibiting a self-focusing effect and reverse saturable absorption, respectively. The second molecular hyperpolarizabilities of the dyad in two conditions were measured to be 3.87 x 10(-30) and 4.82 x 10(-30) esu, respectively. All the results suggest that the azo-ZnPc dyad has potential in the field of nonlinear optics applications. PMID- 18512977 TI - Identification of the molecular motions responsible for the slower secondary (beta) relaxation in sucrose. AB - Broad-band isothermal dielectric relaxation measurements of anhydrous sucrose were made at ambient pressure in its liquid and glassy states. We found a new secondary relaxation that is slower than the one commonly observed in sugars. Additionally, we carried out the dielectric measurements of the equimolar mixture of D-glucose and D-fructose in wide ranges of temperature and frequency. Comparison of the behavior of these two systems allowed us to make suggestions on the origin of the slower beta-relaxation in sucrose. Computer simulations and coupling model calculations were performed to support our interpretation of the kind of molecular motions responsible for the slower secondary relaxation in the disaccharide considered. PMID- 18512978 TI - Nucleation of NaCl nanoparticles in supercritical water: molecular dynamics simulations. AB - Formation of NaCl nanoparticles in supercritical water is studied using molecular dynamics simulation method. We have simulated particle nucleation and growth in NaCl-H2O fluids, with salt concentration of 5.1 wt %, in the temperature and density range of 673-1073 K and 0.17-0.34 g/cm(3), respectively. The cluster size distributions, the size of critical nuclei and cluster lifetimes are reported. The size distribution of emerging clusters shows a very strong dependence on the system's density, with larger clusters forming at lower densities. Clusters consisting of approximately 14-24 ions appear critical for the thermodynamic states examined. The local structures of critical clusters are found to be amorphous. The lifetime values for clusters containing more than 20 ions are in the range of 10-50 ps. We have calculated the NaCl nucleation rates, which appear to be on the order of 10(28) cm(-3) s(-1). PMID- 18512979 TI - Location of magnetic and fluorescent nanoparticles encapsulated inside giant liposomes. AB - In this paper, we demonstrate the production of highly magnetic and fluorescent giant vesicles by encapsulating gamma-Fe2O3-rhodamine B nanoparticles. The liposomes containing the nanoparticles were made of 1,2-dioleoyl-sn-glycero-3 phosphocholine (DOPC). We found that the ionic strength of the initial magnetic fluid is a crucial parameter in controlling the physicochemical properties of the bilayer. At high ionic strength, we obtained very important deformations of liposomes with high magnetic susceptibilities induced by an applied magnetic field. The encapsulation rate was studied using magnetophoresis and photobleaching tests, and the membrane properties were studied using confocal microscopy and elastic measurements. PMID- 18512980 TI - Solvation and aggregation of n,n'-dialkylimidazolium ionic liquids: a multinuclear NMR spectroscopy and molecular dynamics simulation study. AB - The solvation and aggregation of the ionic liquid (IL) 1-n-butyl-3 methylimidazolium chloride ([C4mim]Cl) in water and dimethylsulfoxide (DMSO) were examined by analysis of (1)H and (35/37)Cl chemical shift perturbations and molecular dynamics (MD) simulations. Evidence of aggregation of the IL n-butyl chains in aqueous environments at IL concentrations of 75-80 wt% was observed both in the NMR experiments and in the MD simulations. The studies also show that [C4mim]Cl behaves as a typical electrolyte in water, with both ions completely solvated at low concentrations. On the other hand, the data reveal that the interactions between the [C4mim](+) and Cl(-) ions strengthen as the DMSO content of the solutions increases, and IL-rich clusters persist in this solvent even at concentrations below 10 wt%. These results provide an experimentally supported atomistic explanation of the effects that these two solvents have on some of the macroscopic properties of [C4mim]Cl. The implications that these findings could have on the design of IL-based solvent systems are briefly discussed. PMID- 18512981 TI - Raman spectroscopy of aminated and ultrafine 1,3,5-triamino-2,4,6-trinitrobenzene and PBX 9502 as a function of pressing pressure. AB - The Raman spectra of emulsion aminated, wet aminated, dry aminated, and ultrafine 1,3,5-triamino-2,4,6-trinitrobenzene (TATB) and PBX 9502 explosive powders are measured in a pellet die at pressures from ambient to 180 MPa with a 632.8 nm helium-neon laser. Raman peak frequencies and line widths are calculated from accurately calibrated spectra. The spectral region below 400 cm(-1), where the pressure shifts of the Raman peaks are larger, is emphasized. The most salient effect of pressing is an approximate permanent doubling of the line width of the ambient 56.9 cm(-1) peak for all TATB types. This peak is also much wider in the ultrafine TATB powder than in the other TATB powders, which indicates that the process of creating the ultrafine powder also creates changes in the TATB crystals. The peaks in the spectra of the aminated TATBs and PBX 9502 are very similar, but differences in the fluorescence backgrounds correlate with the expected crystalline purity differences from the different amination processes. The peak frequencies versus pressure for several of the more intense low frequency peaks can be fit well to linear functions between 40 and 180 MPa. The pressure slopes of the emulsion aminated peaks are consistently larger than the slopes of the other powders. Gruneisen parameters calculated from peaks below 100 cm(-1) are scattered, which is probably caused by the anisotropy of TATB crystals and different types of intermolecular bonds. PMID- 18512982 TI - Molecular dynamics simulations of heptyl phosphonic acid: a potential polymer component for fuel cell polymer membrane. AB - Atomistic molecular dynamics simulations have been performed on heptyl phosphonic acid (HPA) to understand the dynamic hydrogen bonding network in the liquid phase. HPA is a phosphonic-acid functionalized alkane (heptane) and a model compound for one of the promising polymers for high temperature (>100 degrees C) fuel cell polymer electrolyte membranes. For the simulation, a force field for this molecule has been generated with the help of quantum chemical calculations and optimized by simplex algorithm. The force field has been validated against experimentally measured properties, for example, density and self-diffusion constant. From molecular dynamics simulations conducted at different temperatures, we have confirmed the hypothesis of dynamic hydrogen bond network formation in this material. PMID- 18512983 TI - Influence of magnesium ions on spontaneous opening of DNA base pairs. AB - A large amount of experimental evidence is available for the effects of magnesium ions on the structure and the stability of the DNA double helix. Less is known, however, on how these ions affect the dynamics of the molecule and the stability of each individual base pair. The present work addresses these questions by a study of the DNA duplex [dCGCAGATCTGCG]2, and its interactions with magnesium ions using nuclear magnetic resonance (NMR) spectroscopy and proton exchange. Two dimensional NMR experiments indicate that binding of magnesium to this DNA duplex does not affect its structure. However, even in the absence of structural changes, magnesium ions specifically affect the exchange properties of imino protons in the four GC/CG base pairs that are located in the interior of the double helix. These specific changes do not result from alterations in the rates of spontaneous opening of these base pairs. Instead, the changes most likely reflect an enhancement in the energetic propensity for spontaneous opening of the GC/CG base pairs that is induced by the binding of magnesium ions. PMID- 18512984 TI - Tenacigenin B derivatives reverse P-glycoprotein-mediated multidrug resistance inHepG2/Dox cells. AB - Tenacissimoside A (1) and 11alpha-O-benzoyl-12beta- O-acetyltenacigenin B (2), two derivatives of tenacigenin B (3) from the plant Marsdenia tenacissima, reversed multidrug resistance in P-glycoprotein (Pgp)-overexpressing multidrug resistant cancer cells. The sensitivity of HepG2/Dox cells to the antitumor drugs doxorubicin, vinblastine, puromycin, and paclitexel was increased by 18-, 10-, 11 , and 6-fold by 20 microg/mL (or 25 microM) of 1 and 16-, 53-, 16-, and 326-fold by 20 microg/mL (or 39 microM) of 2, respectively. A preliminary mechanistic study has suggested that 1 might modulate Pgp-mediated multidrug resistance through directly interacting with the Pgp substrate site. PMID- 18512985 TI - Phloroglucinols with antioxidant activity and xanthonolignoids from the heartwood of Hypericum geminiflorum. AB - A new phloroglucinol, hyperielliptone HA (1/1a), a new spirophloroglucinol possessing an unprecedented skeleton, hyperielliptone HB (2/2a), and two new xanthonolignoids, hyperielliptones HC (3) and HD (4), were isolated from the heartwood of Hypericum geminiflorum. Compounds 1/1a and 2/2a were obtained as tautomeric pairs. The structures and relative configurations of these compounds were elucidated by spectroscopic methods. In biological testing, compound 2/ 2a revealed significant inhibition of oxidative DNA damage and an inhibitory effect on xanthine oxidase. PMID- 18512986 TI - Structural and stereochemical studies of alpha-methylene-gamma-lactone-bearing cembrane diterpenoids from a south china sea soft coral Lobophytum crassum. AB - Four new alpha-methylene-gamma-lactone-bearing cembranoids, 20-acetylsinularolide B (6), presinularolide B (7), 3-dehydroxylpresinularolide B (8), and 3-dehydroxyl 20-acetylpresinularolide B (9), together with five known analogues, sinularolides B-E (1- 4) and 20-acetylsinularolide C (5), were isolated from a South China Sea soft coral Lobophytum crassum. Their structures and relative stereochemistry were established by a combination of detailed spectroscopic data analysis and chemical correlations. The structures of 1- 9 were further confirmed by an X-ray diffraction study on a single crystal of sinularolide B (1). The absolute configurations of sinularolide B (1) and presinularolide B (7) were determined by a novel solid-state CD/TDDFT approach and by a modified Mosher's method, respectively. This study also revealed that the coupling constant between the lactonic methine protons ((3) J 1,2) varies considerably with different functional groups on the cembrane ring and that the determination of the stereochemistry of lactone ring fusion based on this coupling constant is risky. In a bioassay, sinularolides B and C (1 and 2) and new cembranoids 7 and 8 showed in vitro cytotoxicity against the tumor cell lines A-549 and P-388. PMID- 18512987 TI - Alisiaquinones and alisiaquinol, dual inhibitors of Plasmodium falciparum enzyme targets from a New Caledonian deep water sponge. AB - Four new meroterpenes, alisiaquinones A-C (1-3) and alisiaquinol (4), were isolated from a New Caledonian deep water sponge. Their structures and relative stereochemistry were elucidated by spectroscopic data analysis. They are related to xestoquinone, but showed unusual substitution on a tetrahydrofuran junction. They displayed micromolar range activity on two enzymatic targets of importance for the control of malaria, the plasmodial kinase Pfnek-1 and a protein farnesyl transferase, as well as on different chloroquine-sensitive and -resistant strains of Plasmodium falciparum. Alisiaquinone C displayed a submicromolar activity on P. falciparum and a competitive selectivity index on the different plasmodial strains. PMID- 18512988 TI - Quercetin and rutin as potential sunscreen agents: determination of efficacy by an in vitro method. AB - Given that flavonoids are known for their ultraviolet (UV)B photoprotective properties in plants that contain them, we chose to study quercetin (1) and rutin (2) as agents that could potentially be used in sunscreen products. These two substances proved to behave in similar ways. When incorporated in oil-in-water emulsions, at a concentration of 10% (w/w), 1 and 2 give sun protection factor (SPF) values similar to that of homosalate, a standard substance. These two flavonoids also provided a non-negligible level of photoprotection in the UVA range. When used in association with titanium dioxide, the SPF obtained was around 30. PMID- 18512989 TI - Asymmetric syn-selective Henry reaction catalyzed by the sulfonyldiamine-CuCl pyridine system. AB - A catalytic asymmetric Henry reaction has been developed with use of a sulfonyldiamine-CuCl complex as a catalyst. A series of new binaphthyl-containing sulfonyldiamine ligands (2a-h) were readily synthesized in two steps starting from commercially available chiral 1,2-diamines. The (R,R)-diamine-(R)-binaphthyl ligand (2d)-CuCl complex smoothly catalyzed the enantioselective Henry reaction with the assistance of pyridine to give the corresponding adduct with high enantiomeric excess (up to 93%). Moreover, the 2d-CuCl-pyridine system promotes the diastereoselective Henry reaction in syn-selective manner to give the adduct in up to 99% yield with 92:8 syn/ anti selectivity. The enantiomeric excess of the syn-adduct was 84% ee. PMID- 18512990 TI - A facile access to pyrroles from amino acids via an aza-Wacker cyclization. AB - A facile and efficient synthesis of pyrroles from readily available amino acids is described. The key step in the method is an aza-Wacker oxidative cyclization catalyzed by palladium(II)/Cu(OTf)2. A series of pyrroles were obtained by this method under mild conditions. PMID- 18512991 TI - Tuning of chemo- and regioselectivities in multicomponent condensations of 5 aminopyrazoles, dimedone, and aldehydes. AB - Regio- and chemoselective multicomponent protocols for the synthesis of 1,4,6,7,8,9-hexahydro-1H-pyrazolo[3,4-b]quinolin-5-ones, 5,6,7,9 tetrahydropyrazolo[5,1-b]quinazolin-8-ones, and 5a-hydroxy-4,5,5a,6,7,8 hexahydropyrazolo[4,3-c]quinolizin-9-ones starting from 5-amino-3-phenylpyrazole, cyclic 1,3-dicarbonyl compounds and aromatic aldehydes are described. Whereas the three-component coupling in ethanol under reflux conditions provides mixtures of pyrazoloquinolinones and pyrazoloquinazolinones, the condensation can be successfully tuned toward the formation pyrazoloquinolinones (Hantzsch-type dihydropyridines) by performing the reaction at 150 degrees C in the presence of triethylamine base applying sealed vessel microwave or conventional heating. On the other hand, using sonication at room temperature under neutral conditions favors the formation of the isomeric pyrazoloquinazolinones (Biginelli-type dihydropyrimidines). These products are also obtained when the three-component condensation is executed in the presence of trimethylsilylchloride as reaction mediator at high temperatures. A third reaction pathway leading to pyrazoloquinolizinones in a ring-opening/recyclization sequence can be accessed by switching from triethylamine to a more nucleophilic base such as sodium ethoxide or potassium tert-butoxide. The reaction mechanism and intermediates leading to these three distinct tricyclic condensation products are discussed. PMID- 18512992 TI - Easily prepared azopyridines as potent and recyclable reagents for facile esterification reactions. An efficient modified mitsunobu reaction. AB - The 2,2'-, 3,3'-, and 4,4'-azopyridines (azpy) and their alkyl pyridinium ionic liquids were studied as a new class of electron-deficient reagents for Mitsunobu esterification reactions. Among these compounds, 4,4'-azopyridine was found to be the most suitable one for esterification and thioesterification reactions. This new reagent promises to provide general and complementary solutions for separation problems in Mitsunobu reactions without restricting the reaction scope and facilitates the isolation of its hydrazine byproduct. The pyridine hydrazine byproduct can be simply recycled to its azopyridine by an oxidation reaction. PMID- 18512993 TI - Prospect of metal-catalyzed C-C forming cross-coupling reactions in modern solid phase organic synthesis. PMID- 18512994 TI - Temperature-tuning of near-infrared monodisperse quantum dot solids at 1.5 microm for controllable forster energy transfer. AB - We present the first time-resolved cryogenic observations of Forster energy transfer in large, monodisperse lead sulfide quantum dots with ground-state transitions near 1.5 microm (0.8 eV), in environments from 160 K to room temperature. The observed temperature-dependent dipole-dipole transfer rate occurs in the range of (30-50 ns) (-1), measured with our confocal single-photon counting setup at 1.5 microm wavelengths. By temperature-tuning the dots, 94% efficiency of resonant energy transfer can be achieved for donor dots. The resonant transfer rates match well with proposed theoretical models. PMID- 18512995 TI - Opinion: 'Geriatricizing' the health care workforce. PMID- 18512996 TI - Adverse consequences with use of erythropoiesis-stimulating agents in anemia prompt release of guidelines to ensure safe use and maximize benefit. AB - Erythropoietin, a humoral factor produced predominantly in the kidney, stimulates red blood cell production in the bone marrow. Erythropoiesis- stimulating agents (ESAs) have been used for years in the treatment of anemia, with extensive experience and benefits in anemia of chronic kidney disease. Recent data have suggested adverse consequences with use of ESAs, perhaps relating to inappropriate use, and prompting release of guidelines to ensure safe use and maximize benefit. When prescribing ESAs, indications, requirements to monitor laboratory parameters (hemoglobin levels and ferrokinetics), and clinical status need to be stringently followed. PMID- 18512997 TI - A practical approach to guide clinicians in the evaluation of male patients with breast masses. AB - Breast cancer must be considered in the evaluation of breast masses in men, although various benign causes are more common, including gynecomastia and conditions of the skin and subcutaneous tissue. A patient's history may identify key features suspicious for malignancy or reassuring for benign disease. Physical examination has been documented to be as effective as mammography in distinguishing benign from malignant lesions, and both have been reported as highly accurate for the identification of malignancy. Mammography is therefore best used when the physical examination findings are indeterminate. Ultrasonography may be used as an adjunct to mammography; no evidence supports the use of magnetic resonance imaging in male breast patients. If clinical or mammographic features are suspicious or indeterminate for malignancy, tissue diagnosis is warranted and may be achieved surgically or via core-needle biopsy or fine-needle aspiration cytology. Given the lack of uniformity in the clinical recommendations for the evaluation of breast masses in men, a practical approach is proposed. PMID- 18512998 TI - Treating depression in terminally ill patients can optimize their physical comfort at the end of life and provide them the opportunity to confront and prepare for death. AB - The dying process is characterized by feelings of sadness and fear. It is normal for patients at the end of life to worry and grieve the loss of their health. However, when these feelings become excessive and interfere with all aspects of the patient's life, they are abnormal responses to the stress of terminal illness. Screening for depression in terminally ill patients can optimize their physical comfort at the end of life and provide them the opportunity to confront and prepare for death. PMID- 18512999 TI - Case: Asymptomatic, slowly enlarging facial lesion. PMID- 18513000 TI - Quenching of local magnetic moment in oxygen adsorbed graphene nanoribbons. AB - The electronic and magnetic properties of oxidized zigzag and armchair graphene nanoribbons, with hydrogen passivated edges, have been investigated from ab initio pseudopotential calculations within the density functional scheme. The oxygen molecule in its triplet state is adsorbed most stably at the edge of a zigzag nanoribbon. The Stoner metallic behavior of the ferromagnetic nanoribbons and the Slater insulating (ground state) behavior of the antiferromagnetic nanoribbons remain intact upon oxygen adsorption. The formation of a spin-paired C-O bond drastically reduces the local atomic magnetic moment of carbon at the edge of the ferromagnetic zigzag ribbon. PMID- 18513001 TI - Photoelectron spectroscopy of anions at 118.2 nm: observation of high electron binding energies in superhalogens MCl4- (M=Sc, Y, La). AB - High energy photon is needed for photoelectron spectroscopy (PES) of anions with high electron binding energies, such as superhalogens and O-rich metal oxide clusters. The highest energy photon used for anion PES in the laboratory has been 157 nm (7.866 eV) from F2 eximer lasers. Here, we report an anion PES experiment using coherent vacuum ultraviolet radiation at 118.2 nm (10.488 eV) by tripling the third harmonic output (355 nm) of a Nd:YAG laser in a XeAr cell. Our study focuses on a set of superhalogen species, MCl(4) (-) (M=Sc, Y, La), which were expected to possess very high electron binding energies. While the 157 nm photon can only access the ground state detachment features for these species, more transitions to the excited states at binding energies higher than 8 eV are observed at 118.2 nm. The adiabatic detachment energies are shown to be, 6.84, 7.02, and 7.03 eV for ScCl(4) (-), YCl(4) (-), and LaCl(4) (-) eV, respectively, whereas their corresponding vertical detachment energies are measured to be 7.14, 7.31, and 7.38 eV. PMID- 18513002 TI - Impedance measurements on a DNA junction. AB - 1 microm double-stranded DNA molecules are immobilized between pairs of gold and pairs of platinum microelectrodes with gaps of 0.4 and 1 microm, respectively, and their electrical characteristics are determined under the application of constant and sinusoidal bias voltages. Due to their extremely high impedance for constant voltage bias, the samples of DNA are excellent insulators; however, their impedances show strong frequency dependence in the range of 10 Hz-7.5 MHz. Favorable response in the gold electrodes is attributed to the higher ability of DNA molecules to bridge the narrower gold electrode gaps in contrast to that in the wider platinum junctions. PMID- 18513003 TI - Resolutions of the Coulomb operator. AB - We discuss a generalization of the resolution of the identity by considering one body resolutions of two-body operators, with particular emphasis on the Coulomb operator. We introduce a set of functions that are orthonormal with respect to 1r(12) and propose that the resulting "resolution of the Coulomb operator," r(12) (-1)=mid R:phi(i)phi(i)mid R:, may be useful for the treatment of large systems due to the separation of two-body interactions. We validate our approach by using it to compute the Coulomb energy of large systems of point charges. PMID- 18513004 TI - Virial exchange energies from model exact-exchange potentials. AB - It is shown by the example of Slater's averaged exchange potential that a poor approximation to the optimized effective potential (OEP) can yield a deceptively accurate energy via the conventional Kohn-Sham energy functional. For a trial exchange potential to be correct, its Kohn-Sham energy must coincide with the value obtained by the Levy-Perdew virial relation. Significant discrepancies between Kohn-Sham and the virial exchange energies are found for self-consistent Slater, Becke-Johnson, and effective local potentials (ELPs); their relative magnitudes are used to argue that, as approximations to the exact-exchange OEP, ELPs are the most accurate. Virial energy discrepancies vanish for Yang-Wu OEPs when the orbital and auxiliary basis sets are balanced, and remain surprisingly small for oscillatory OEPs obtained with unbalanced basis sets. PMID- 18513005 TI - An application of the van der Waals density functional: Hydrogen bonding and stacking interactions between nucleobases. AB - We apply the van der Waals density functional (vdW-DF) to study hydrogen bonding and stacking interactions between nucleobases. The excellent agreement of our results with high level quantum chemical calculations highlights the value of the vdW-DF for first-principles investigations of biologically important molecules. Our results suggest that, in the case of hydrogen-bonded nucleobase pairs, dispersion interactions reduce the cost of propeller twists while having a negligible effect on buckling. Furthermore, the efficient scaling of DFT methods allowed for the easy optimization of separation distance between nucleobase stacks, indicating enhancements in the interaction energy of up to 3 kcalmol over previous fixed distance calculations. We anticipate that these results are significant for extending the vdW-DF method to model larger vdW complexes and biological molecules. PMID- 18513006 TI - Spectroscopic data for the LiH molecule from pseudopotential quantum Monte Carlo calculations. AB - Quantum Monte Carlo and quantum chemistry techniques are used to investigate pseudopotential models of the lithium hydride (LiH) molecule. Interatomic potentials are calculated and tested by comparing with the experimental spectroscopic constants and well depth. Two recently developed pseudopotentials are tested, and the effects of introducing a Li core polarization potential are investigated. The calculations are sufficiently accurate to isolate the errors from the pseudopotentials and core polarization potential. Core-valence correlation and core relaxation are found to be important in determining the interatomic potential. PMID- 18513007 TI - Equivalence of two approaches for quantum-classical hybrid systems. AB - We discuss two approaches that are frequently used to describe quantum-classical hybrid system. One is the well-known mean-field theory and the other adopts a set of hybrid brackets which is a mixture of quantum commutators and classical Poisson brackets. We prove that these two approaches are equivalent. PMID- 18513008 TI - An efficient algorithm for solving nonlinear equations with a minimal number of trial vectors: applications to atomic-orbital based coupled-cluster theory. AB - The conjugate residual with optimal trial vectors (CROP) algorithm is developed. In this algorithm, the optimal trial vectors of the iterations are used as basis vectors in the iterative subspace. For linear equations and nonlinear equations with a small-to-medium nonlinearity, the iterative subspace may be truncated to a three-dimensional subspace with no or little loss of convergence rate, and the norm of the residual decreases in each iteration. The efficiency of the algorithm is demonstrated by solving the equations of coupled-cluster theory with single and double excitations in the atomic orbital basis. By performing calculations on H(2)O with various bond lengths, the algorithm is tested for varying degrees of nonlinearity. In general, the CROP algorithm with a three-dimensional subspace exhibits fast and stable convergence and outperforms the standard direct inversion in iterative subspace method. PMID- 18513009 TI - Determination of free energy profiles by repository based adaptive umbrella sampling: bridging nonequilibrium and quasiequilibrium simulations. AB - We propose a new adaptive sampling approach to determine free energy profiles with molecular dynamics simulations, which is called as "repository based adaptive umbrella sampling" (RBAUS). Its main idea is that a sampling repository is continuously updated based on the latest simulation data, and the accumulated knowledge and sampling history are then employed to determine whether and how to update the biasing umbrella potential for subsequent simulations. In comparison with other adaptive methods, a unique and attractive feature of the RBAUS approach is that the frequency for updating the biasing potential depends on the sampling history and is adaptively determined on the fly, which makes it possible to smoothly bridge nonequilibrium and quasiequilibrium simulations. The RBAUS method is first tested by simulations on two simple systems: a double well model system with a variety of barriers and the dissociation of a NaCl molecule in water. Its efficiency and applicability are further illustrated in ab initio quantum mechanics/molecular mechanics molecular dynamics simulations of a methyl transfer reaction in aqueous solution. PMID- 18513010 TI - A stereographic projection path integral study of the coupling between the orientation and the bending degrees of freedom of water. AB - A Monte Carlo path integral method to study the coupling between the rotation and bending degrees of freedom for water is developed. It is demonstrated that soft internal degrees of freedom that are not stretching in nature can be mapped with stereographic projection coordinates. For water, the bending coordinate is orthogonal to the stereographic projection coordinates used to map its orientation. Methods are developed to compute the classical and quantum Jacobian terms so that the proper infinitely stiff spring constant limit is recovered in the classical limit, and so that the nonconstant nature of the Riemann Cartan curvature scalar is properly accounted in the quantum simulations. The theory is used to investigate the effects of the geometric coupling between the bending and the rotating degrees of freedom for the water monomer in an external field in the 250 to 500 K range. We detect no evidence of geometric coupling between the bending degree of freedom and the orientations. PMID- 18513011 TI - Universal mathematical identities in density functional theory: results from three different spin-resolved representations. AB - This paper supersedes previous theoretical approaches to conceptual DFT because it provides a unified and systematic approach to all of the commonly considered formulations of conceptual DFT, and even provides the essential mathematical framework for new formulations. Global, local, and nonlocal chemical reactivity indicators associated with the "closed-system representation" ([N(alpha),N(beta),nu(alpha)(r),nu(beta)(r)]) of spin-polarized density functional theory (SP-DFT) are derived. The links between these indicators and the ones associated with the "open-system representation" ([mu(alpha),mu(beta),nu(alpha)(r),nu(beta)(r)]) are derived, including the spin resolved Berkowitz-Parr identity. The Legendre transform to the "density representation" ([rho(alpha)(r),rho(beta)(r)]) is performed, and the spin resolved Harbola-Chattaraj-Cedillo-Parr identities linking the density representation to the closed-system and open-system representations are derived. Taken together, these results provide the framework for understanding chemical reactions from both the electron-following perspective (using either the closed system or the open-system representation) and electron-preceding perspective (density representation). A powerful matrix-vector notation is developed; with this notation, identities in conceptual DFT become universal. Specifically, this notation allows the fundamental identities in conventional (spin-free) conceptual DFT, the [N(alpha),N(beta)] representation, and the [N=N(alpha)+N(beta),N(S)=N(alpha)-N(beta)] representation to be written in exactly the same forms. In cases where spin transfer and electron transfer are coupled (e.g., radical+molecule reactions), we believe that the [N(alpha),N(beta)] representation may be more useful than the more common [N,N(S)] representation. PMID- 18513012 TI - The restricted active space followed by second-order perturbation theory method: theory and application to the study of CuO2 and Cu2O2 systems. AB - A multireference second-order perturbation theory using a restricted active space self-consistent field wave function as reference (RASPT2/RASSCF) is described. This model is particularly effective for cases where a chemical system requires a balanced orbital active space that is too large to be addressed by the complete active space self-consistent field model with or without second-order perturbation theory (CASPT2 or CASSCF, respectively). Rather than permitting all possible electronic configurations of the electrons in the active space to appear in the reference wave function, certain orbitals are sequestered into two subspaces that permit a maximum number of occupations or holes, respectively, in any given configuration, thereby reducing the total number of possible configurations. Subsequent second-order perturbation theory captures additional dynamical correlation effects. Applications of the theory to the electronic structure of complexes involved in the activation of molecular oxygen by mono- and binuclear copper complexes are presented. In the mononuclear case, RASPT2 and CASPT2 provide very similar results. In the binuclear cases, however, only RASPT2 proves quantitatively useful, owing to the very large size of the necessary active space. PMID- 18513013 TI - Differential cross sections and product energy distributions for the C(3P)+OH(X 2Pi)-->CO(X 1Sigma+)+H(2S) reaction using a quasiclassical trajectory method. AB - Quasiclassical trajectory calculations have been carried out for the C((3)P)+OH(X (2)Pi)-->CO(X (1)Sigma(+))+H((2)S) reaction using a recent ab initio potential energy surface for the ground electronic state X (2)A(') of COH. Differential cross sections (DCSs), and product vibrational, rotational and translational distributions have been determined for a wide range of collision energies (0.001 1 eV). The role of excitations (rotation or vibration) of the OH reactant on these quantities has been investigated. Product vibrational, rotational, and translational distributions are found to be almost independent on the rovibrational state of OH, whereas DCSs show a weak dependence on the initial rotational state of OH. We also analyze the results using a study based on the lifetime of the intermediate complex and on the kinematic constraint associated with the mass combination. PMID- 18513014 TI - Ultrafast strong-field dissociative ionization dynamics of CH2Br2 probed by femtosecond soft x-ray transient absorption spectroscopy. AB - Femtosecond time-resolved soft x-ray transient absorption spectroscopy based on a high-order harmonic generation source is used to investigate the dissociative ionization of CH(2)Br(2) induced by 800 nm strong-field irradiation. At moderate laser peak intensities (2.0 x 10(14) Wcm(2)), strong-field ionization is accompanied by ultrafast C-Br bond dissociation, producing both neutral Br ((2)P(32)) and Br(*) ((2)P(12)) atoms together with the CH(2)Br(+) fragment ion. The measured rise times for Br and Br(*) are 130+/-22 fs and 74+/-10 fs, respectively. The atomic bromine quantum state distribution shows that the BrBr(*) population ratio is 8.1+/-3.8 and that the Br (2)P(32) state is not aligned. The observed product distribution and the time scales of the photofragment appearances suggest that multiple field-dressed potential energy surfaces are involved in the dissociative ionization process. At higher laser peak intensities (6.2 x 10(14) Wcm(2)), CH(2)Br(2) (+) undergoes sequential ionization to form the metastable CH(2)Br(2) (2+) dication. These results demonstrate the potential of core-level probing with high-order harmonic transient absorption spectroscopy for studying ultrafast molecular dynamics. PMID- 18513015 TI - Circular dichroism spectroscopy of small molecular aggregates: dynamical features and size effects. AB - Circular dichroism (CD) spectra of small molecular aggregates, taking monomer vibrational motion into account, are calculated by employing wave packet propagation techniques. The spectra are related to the population and exciton transfer in the aggregates. It is investigated as to how far this dynamics can be directly extracted from the CD spectra. The method is applied to aggregates of perylene bisimides where temperature dependent spectra have recently been recorded in our laboratory [V. Dehm et al., Org. Lett. 9, 1085 (2007)]. PMID- 18513016 TI - Interatomic contributions to high-energy electron-molecule scattering. AB - Within the independent atom scattering model, we derive an approximate formula for the rotationally and vibrationally averaged three-atom terms in the series expansion of the electron scattering cross section. This formula uses the atomic scattering factors as well as the atomic scattered wavefunctions as input, and rotational averaging is performed numerically. We compare our results to previous theoretical multiple scattering approaches for the molecules F(3), F(4), and SF(6) and to experimental data for TeF(6). Our results are consistent with those of previous calculations and inclusion of the three-atom term produces a dramatically better least squares fit of the TeF(6) data. The algorithm presented here is sufficiently fast and simple to be incorporated easily into existing electron diffraction codes. PMID- 18513017 TI - An investigation of the molecular geometry and electronic structure of nitryl chloride by a combination of rotational spectroscopy and ab initio calculations. AB - The ground-state rotational spectra of the six isotopomers (16)O(2) (14)N(35)Cl, (16)O(2) (14)N(37)Cl, (18)O(16)O(14)N(35)Cl, (18)O(2) (14)N(35)Cl, (16)O(2) (15)N(35)Cl, and (16)O(2) (15)N(37)Cl of nitryl chloride were observed with a pulsed-jet, Fourier-transform microwave spectrometer to give rotational constants, Cl and (14)N nuclear quadrupole coupling, and spin-rotation coupling constants. These spectroscopic constants were interpreted to give r(0), r(s), and r(m) ((2)) versions of the molecular geometry and information about the electronic redistribution at N when nitryl chloride is formed from NO(2) and a Cl atom. The r(m) ((2)) geometry has r(N-Cl)=1.8405(6) A, r(N-O)=1.1929(2) A, and the angle ONO=131.42(4) degrees , while the corresponding quantities for the r(s) geometry are 1.8489 A, 1.1940 A, and 131.73 degrees , respectively. Electronic structure calculations at CCSD(T)cc-pVXZ (X=T, Q, or 5) levels of theory were carried out to give a r(e) geometry, vibration-rotation corrections to equilibrium rotational constants, and values of the (35)Cl and (14)N nuclear hyperfine (quadrupole and spin-rotation) coupling constants in good agreement with experiment. The equilibrium geometry at the CCSD(T)/cc-pV5Z level of theory has r(N-Cl)=1.8441 A, r(N-O)=1.1925 A and the angle ONO=131.80 degrees . The observed rotational constants were corrected for the vibration-rotation effects calculated ab initio to yield semiempirical equilibrium constants which were then fitted to give the following semiempirical equilibrium geometry: r(N Cl)=1.8467(2) A, r(N-O)=1.1916(1) A, and the angle ONO=131.78(3) degrees . PMID- 18513018 TI - Determination of the rate constant for sulfur recombination by quasiclassical trajectory calculations. AB - The sulfur recombination reaction has been thought of as one of the most important chemical reactions in the volcanic activities of the planet. It is also important in determining the propagation of elemental sulfur in the atmosphere. There have been two experimental attempts to determine the reaction rate of the S+S-->S(2) recombination, however their results differ by four orders of magnitude. In this work, we determine the rate constant of S+S-->S(2) from quasiclassical trajectory calculations. The third order rate constant at 298.15 K predicted by the present calculations is 4.19 x 10(-33) cm(6) molecules(-2) s( 1), which is in excellent agreement with the determination of Fair and Thrush [Trans. Faraday Soc. 65, 1208 (1969)]. The temperature dependent rate constant is determined to be 3.94 x 10(-33) exp[205.56(1T-1298.15)], which was determined from the temperature range of 100-500 K. PMID- 18513019 TI - Quantum capture, adiabatic channel, and classical trajectory study of the high pressure rate constant of the reaction H+O2-->HO2 between 0 and 5000 K. AB - Limiting high pressure rate constants for the recombination reaction H+O(2)- >HO(2) are modeled between 0 and 5000 K on an ab initio potential. Quantum capture theory is employed for the temperature range from 0 to about 1 K, while classical trajectory calculations are suitable for covering temperatures above about 200 K. The intermediate temperature range is analyzed by adiabatic channel capture theory. The system is characterized by transition-state switching from outer transition states in the long-range-C(6)R(6) potential to inner transition states in the range of a "shoulder" of the potential. The limiting high pressure rate constants from the trajectory calculations are sufficient for comparison with the experimental data which are available over the range from 300 to 900 K. Specific rate constants k(E,J) for HO(2) dissociation are also given and analyzed with respect to internal consistency with capture cross sections. PMID- 18513020 TI - Laser induced fluorescence spectroscopy of NC3O. AB - Laser induced fluorescence spectra of the NC(3)O radical in a supersonic jet have been observed. The radical was produced in a pulsed electric discharge of HC(3)N and O(2) diluted to 0.3% with Ar. A total of 17 vibronic bands with a radiative lifetime of approximately 30 ns have been observed in a region from 27 000 to 27 500 cm(-1). The observed vibronic bands are classified as (2)Pi(12)-(2)Pi(12), (2)Pi(32)-(2)Pi(32), and (2)Sigma-(2)Sigma types. The upper states of the (2)Sigma-(2)Sigma bands have large spin-rotation constants, which should be denoted as Sigma((+)) and Sigma((-)). From high-level ab initio calculations and rotational analyses, the observed transition was assigned to the B (2)Pi-X (2)A(") transition. Dispersed fluorescence spectra from the upper (2)Sigma and (2)Pi vibronic levels have also been observed, yielding fundamental vibrational frequencies for the nu(1), nu(2), nu(3), and nu(7) modes of the ground state. PMID- 18513021 TI - Study of the Xe-NH3 van der Waals complex: high-resolution microwave spectra and ab initio calculations. AB - An ab initio potential energy surface of the Xe-NH(3) van der Waals complex was constructed at the coupled cluster level of theory with single, double, and pertubatively included triple excitations. The small-core pseudopotential and augmented correlation-consistent polarized valence quadruple-zeta basis set was used for the Xe atom and Dunning's augmented correlation-consistent polarized valence triple-zeta basis set for the other atoms. The basis sets were supplemented with midbond functions. Rotational spectra of the Xe-NH(3) van der Waals complex were recorded using a pulsed-nozzle Fourier transform microwave spectrometer. Rotational transitions within two internal rotor states, namely, the Sigma0(0) and Pi1(1) (lower) states, were measured and assigned to the Xe (14)NH(3) and Xe-(15)NH(3) isotopologues. For the deuterated isotopologues, only the Sigma0(0) states were observed. Two inversion components were observed for each state except for the "s" component of the Sigma0(0) state of the Xe (14)NH(3) and Xe-(15)NH(3) isotopologues, which has a spin statistical weight of zero. Nuclear quadrupole hyperfine structures arising from the (14)N (nuclear spin angular momentum quantum number I=1) and (131)Xe (I=32) nuclei were detected and analyzed. The observed spectra suggest that the Pi1(1) (lower) state has lower energy than the unobserved Sigma1(1) state, in contrast to the case of Ar NH(3). PMID- 18513022 TI - The theoretical prediction of infrared spectra of trans- and cis-hydroxycarbene calculated using full dimensional ab initio potential energy and dipole moment surfaces. AB - Accurate infrared spectra of the two hydroxycarbene isomers are computed by diagonalizing the Watson Hamiltonian including up to four mode couplings using full dimensional potential energy and dipole moment surfaces calculated at the CCSD(T)/cc-pVTZ (frozen core) and CCSD6-311G(**) (all electrons correlated) levels, respectively. Anharmonic corrections are found to be very important for these elusive higher-energy isomers of formaldehyde. Both the energy levels and intensities of stretching fundamentals and all overtone transitions are strongly affected by anharmonic couplings between the modes. The results for trans HCOHHCOD are in excellent agreement with the recently reported IR spectra, which validates our predictions for the cis-isomers. PMID- 18513023 TI - Femtosecond time-resolved photoelectron-photoion coincidence imaging of multiphoton multichannel photodynamics in NO2. AB - The multiphoton multichannel photodynamics of NO(2) has been studied using femtosecond time-resolved coincidence imaging. A novel photoelectron-photoion coincidence imaging machine was developed at the laboratory in Amsterdam employing velocity map imaging and "slow" charged particle extraction using additional electron and ion optics. The NO(2) photodynamics was studied using a two color pump-probe scheme with femtosecond pulses at 400 and 266 nm. The multiphoton excitation produces both NO(2) (+) parent ions and NO(+) fragment ions. Here we mainly present the time dependent photoelectron images in coincidence with NO(2) (+) or NO(+) and the (NO(+),e) photoelectron versus fragment ion kinetic energy correlations. The coincidence photoelectron spectra and the correlated energy distributions make it possible to assign the different dissociation pathways involved. Nonadiabatic dynamics between the ground state and the A (2)B(2) state after absorption of a 400 nm photon is reflected in the transient photoelectron spectrum of the NO(2) (+) parent ion. Furthermore, Rydberg states are believed to be used as "stepping" states responsible for the rather narrow and well-separated photoelectron spectra in the NO(2) (+) parent ion. Slow statistical and fast direct fragmentation of NO(2) (+) after prompt photoelectron ejection is observed leading to formation of NO(+)+O. Fragmentation from both the ground state and the electronically excited a (3)B(2) and b (3)A(2) states of NO(2) (+) is observed. At short pump probe delay times, the dominant multiphoton pathway for NO(+) formation is a 3x400 nm+1x266 nm excitation. At long delay times (>500 fs) two multiphoton pathways are observed. The dominant pathway is a 1x400 nm+2x266 nm photon excitation giving rise to very slow electrons and ions. A second pathway is a 3x400 nm photon absorption to NO(2) Rydberg states followed by dissociation toward neutral electronically and vibrationally excited NO(A (2)Sigma,v=1) fragments, ionized by one 266 nm photon absorption. As is shown in the present study, even though the pump-probe transients are rather featureless the photoelectron-photoion coincidence images show a complex time varying dynamics in NO(2). We present the potential of our novel coincidence imaging machine to unravel in unprecedented detail the various competing pathways in femtosecond time-resolved multichannel multiphoton dynamics of molecules. PMID- 18513024 TI - Lifetime measurements of SO2 below the Clement's A-band. AB - High resolution laser induced fluorescence spectrum of jet-cooled SO(2) is recorded toward the blue side of the Clement's A-Band in the region of 314-319 nm. Time resolved fluorescence measurements have been carried out for all the prominent peaks in this region. Most of the peaks exhibited double exponential decay profiles. Some of the rovibronic bands exhibited quantum beats with strong quantum beats observed at 315.261 and 315.271 nm. This is the first observation of quantum beats in SO(2) in the absence of any external magnetic or electric fields. The decay profiles of the beating rovibronic bands were fitted using a four-level model by least-squares fitting method. The fitting shows that all the measured bands were double exponential with a similar first lifetime of approximately 3 mus and a varying second lifetime of the order of 1 micros-100 ns with a beating frequency of approximately 1 MHz. These quantum beats, in the absence of any external field, indicate rotational level mixing between the A (1)A(2) and the B (1)B(1) vibronic states which are near resonant due to the high density of states of these two states. PMID- 18513025 TI - Observation and calculation of the Cs2 2 3Delta(1g) and b 3Pi(0u) states. AB - The Cs(2) 2 (3)Delta(1g) and b (3)Pi(0u) states have been observed by infrared infrared double resonance spectroscopy for the first time. 221 2 (3)Delta(1g)<--A (1)Sigma(u) (+)<--X (1)Sigma(g) (+) double resonance lines have been assigned to transitions into the 2 (3)Delta(1g) v=6-13 vibrational levels. Resolved fluorescence into the b (3)Pi(0u) v(')=0-48 levels has been recorded. Molecular constants and potential energy curves are determined by the global fit of the entire set of the experimental data. Theoretical potential energy curves of the 2 (3)Delta(g) and b (3)Pi(u) states have been determined in the framework of the pseudopotential method and are compared with the experimental results. PMID- 18513026 TI - A plastic phase of water from computer simulation. AB - We report a member of ices called plastic or rotator phase, in which individual water molecules make facile rotations as in liquid state but are held tightly in an ordered structure. Molecular dynamics simulations of three classical models of water show that a plastic ice phase appears at a temperature when ice VII is heated or liquid water is cooled at high pressures above several gigapascals. A large amount of latent heat is absorbed when ice VII is transformed to the rotator phase at 590 K and 10 GPa, which is a typical characteristic of the plastic transitions for nearly spherical molecules. In addition to the spontaneous formation of plastic phase in the simulations, its existence is supported by robustness of plastic phase for hypothetical water with varying degrees of Coulombic interactions. PMID- 18513027 TI - A neutron scattering study of strong-symmetric hydrogen bonds in potassium and cesium hydrogen bistrifluoroacetates: Determination of the crystal structures and of the single-well potentials for protons. AB - The crystal structures of potassium and cesium bistrifluoroacetates, KH(CF(3)COO)(2) and CsH(CF(3)COO)(2), respectively, were determined at room and cryogenic temperatures with the single crystal neutron diffraction technique. The crystals belong to the monoclinic space groups, I2a and A2a, respectively, and there is no evidence of any structural phase transition. In both crystals, trifluoroacetate entities in centrosymmetric dimers are linked by very short hydrogen bonds lying across a center of inversion. The thermal parameters provide no evidence of any double minimum potential for hydrogen bond protons. Single minimum potentials were determined via best fitting to the inelastic neutron scattering spectral profiles of the stretching vibrations. They comprise a narrow well for the ground state and a very broad quasiharmonic well for excited states. The spread out of the wave functions of these states shows that protons are no longer confined between the oxygens. Presumably, they are attracted by the lone pairs of oxygen atoms. These potentials emphasize the covalent nature of the OO bond and the ionic character of the hydrogen bond proton. PMID- 18513028 TI - Measuring distances between half-integer quadrupolar nuclei and detecting relative orientations of quadrupolar and dipolar tensors by double-quantum homonuclear dipolar recoupling nuclear magnetic resonance experiments. AB - We studied the possibility of using double-quantum homonuclear dipolar recoupling magic angle spinning nuclear magnetic resonance experiments for structural analysis of systems of half-integer quadrupolar nuclei. We investigated symmetry based recoupling schemes R2(2) (1) and R2(2) (1)R2(2) (-1) and showed that the obtained double-quantum filtered signals depend substantially on magnitudes and relative orientations of dipolar and quadrupolar tensors. Experimental results measured on aluminophosphate molecular sieve AlPO(4)-14, containing dipolar coupled spin-52 aluminum nuclei, were compared to results of time-consuming numerical simulations. The comparison for short mixing times allowed us to roughly measure internuclear Al-Al distances, if constraints about relative tensor orientations were available. Inspection of relative orientations of dipolar and quadrupolar tensors, using known distances between nuclei, required experimental and simulated data for long mixing times and yielded less accurate results. Two experimental protocols were employed for measuring double-quantum filtered curves, the symmetric protocol, in which excitation and reconversion periods are incremented simultaneously, and the asymmetric protocol, in which only the length of the excitation period is incremented and the length of the reconversion period is kept constant. The former experimental protocol was more convenient for the detection of internuclear distances, and the latter one was more appropriate for the inspection of relative orientations of interaction tensors. PMID- 18513029 TI - The influence of bond rigidity and cluster diffusion on the self-diffusion of hard spheres with square well interaction. AB - Hard spheres interacting through a square well potential were simulated by using two different methods: Brownian cluster dynamics (BCD) and event driven Brownian dynamics (EDBD). The structure of the equilibrium states obtained by both methods was compared and found to be almost identical. Self-diffusion coefficients (D) were determined as a function of the interaction strength. The same values were found by using BCD or EDBD. Contrary to EDBD, BCD allows one to study the effect of bond rigidity and hydrodynamic interaction within the clusters. When the bonds are flexible, the effect of attraction on D is relatively weak compared to systems with rigid bonds. D increases first with increasing attraction strength, and then decreases for stronger interaction. Introducing intracluster hydrodynamic interaction weakly increases D for a given interaction strength. Introducing bond rigidity causes a strong decrease in D which no longer shows a maximum as function of the attraction strength. PMID- 18513030 TI - Taking apart the two-dimensional infrared vibrational echo spectra: more information and elimination of distortions. AB - Ultrafast two-dimensional infrared (2D-IR) vibrational echo spectroscopy can probe the fast structural evolution of molecular systems under thermal equilibrium conditions. Structural dynamics are tracked by observing the time evolution of the 2D-IR spectrum, which is caused by frequency fluctuations of vibrational mode(s) excited during the experiment. However, there are a variety of effects that can produce line shape distortions and prevent the correct determination of the frequency-frequency correlation function (FFCF), which describes the frequency fluctuations and connects the experimental observables to a molecular level depiction of dynamics. In addition, it can be useful to analyze different parts of the 2D spectrum to determine if dynamics are different for subensembles of molecules that have different initial absorption frequencies in the inhomogeneously broadened absorption line. Here, an important extension to a theoretical method for extraction of the FFCF from 2D-IR spectra is described. The experimental observable is the center line slope (CLSomega(m)) of the 2D-IR spectrum. The CLSomega(m) is obtained by taking slices through the 2D spectrum parallel to the detection frequency axis (omega(m)). Each slice is a spectrum. The slope of the line connecting the frequencies of the maxima of the sliced spectra is the CLSomega(m). The change in slope of the CLSomega(m) as a function of time is directly related to the FFCF and can be used to obtain the complete FFCF. CLSomega(m) is immune to line shape distortions caused by destructive interference between bands arising from vibrational echo emission, from the 0-1 vibrational transition (positive), and from the 1-2 vibrational transition (negative) in the 2D-IR spectrum. The immunity to the destructive interference enables the CLSomega(m) method to compare different parts of the bands as well as comparing the 0-1 and 1-2 bands. Also, line shape distortions caused by solvent background absorption and finite pulse durations do not affect the determination of the FFCF with the CLSomega(m) method. The CLSomega(m) can also provide information on the cross correlation between frequency fluctuations of the 0-1 and 1-2 vibrational transitions. PMID- 18513031 TI - X-ray absorption spectra of hexagonal ice and liquid water by all-electron Gaussian and augmented plane wave calculations. AB - Full potential x-ray spectroscopy simulations of hexagonal ice and liquid water are performed by means of the newly implemented methodology based on the Gaussian augmented plane waves formalism. The computed spectra obtained within the supercell approach are compared to experimental data. The variations of the spectral distribution determined by the quality of the basis set, the size of the sample, and the choice of the core-hole potential are extensively discussed. The second part of this work is focused on the understanding of the connections between specific configurations of the hydrogen bond network and the corresponding contributions to the x-ray absorption spectrum in liquid water. Our results confirm that asymmetrically coordinated molecules, in particular, those donating only one or no hydrogen bond, are associated with well identified spectral signatures that differ significantly from the ice spectral profile. However, transient local structures, with half formed hydrogen bonds, may still give rise to spectra with dominant postedge contributions and relatively weaker oscillator strengths at lower energy. This explains why by averaging the spectra over all the O atoms of liquid instantaneous configurations extracted from ab initio molecular dynamics trajectories, the spectral features indicating the presence of weak or broken hydrogen bonds turn out to be attenuated and sometimes not clearly distinguishable. PMID- 18513032 TI - Molecular theory of barycentric velocity: monatomic fluids. AB - The notion of barycentric velocity appears in irreversible thermodynamics and fluid mechanics, in which it is a field variable obeying the hydrodynamic equations or, more specifically, the momentum balance equation, which is coupled to the rest of hydrodynamic equations. Therefore, its behavior is not known until the hydrodynamic equations are solved for the flow problem of interest. Unlike diffusion fluxes, heat fluxes, or stresses, it does not have its own constitutive relation similar to Fick's law, Fourier's law, and Newton's law of viscosity. In this work, the constitutive equation is derived for it. In parallel to the phenomenological notion of barycentric velocity, the notion of mean fluid velocity appears in statistical mechanics of irreversible dynamic processes according to the theory of Irving and Kirkwood [J. Chem. Phys. 18, 817 (1950)], and plays the same role of the phenomenological counterpart. In this work, we investigate the statistical mechanical meanings of the mean fluid velocity of a fluid in flow beyond its formal connection with the barycentric velocity. We show that it consists of two components; the center-of-gravity velocity of the packet of fluid molecules, which may be identified with the barycentric velocity in the phenomenological theory, and the diffusive contribution of its collective modes relative to the center of gravity. If the fluid is uniform in space or if the packet of fluid mass is rigid, the diffusive component vanishes. The statistical mechanical (molecular theory) formula for the mean fluid velocity provides the constitutive relation for it in terms of density and temperature gradients present in the fluid in flow. The constitutive relation obtained for the mean fluid velocity can be an important component in the theory of transport processes in liquids. Its significance to fluid mechanics is briefly discussed. PMID- 18513033 TI - Discontinuously propagating waves in the bathoferroin-catalyzed Belousov Zhabotinsky reaction incorporated into a microemulsion. AB - Three new types of discontinuously propagating waves are reported in the bathoferroin-catalyzed Belousov-Zhabotinsky (BZ) reaction dispersed in water-in oil Aerosol OT microemulsion. Jumping waves (JWs) are typically observed at or above room temperature and develop from the familiar trigger waves. Bubble waves (BWs) typically emerge from trigger or JWs at similar temperatures, while rotating waves (RWs) evolve from JW at higher temperatures (>40 degrees C). All these waves propagate discontinuously in a saltatory fashion. Other characteristic features include a discontinuous front for BW consisting of small concentric waves (bubbles) and lateral rotation of annular RWs. All three types of waves, as well as segmented but continuously propagating waves, can coexist. A simple model that is able to describe both jumping and segmented waves is described. PMID- 18513034 TI - Infrared absorption spectra of vinyl radicals isolated in solid Ne. AB - Irradiation of samples of solid Ne near 3.0 K containing ethene (C(2)H(4)) with vacuum ultraviolet radiation at 120 nm from synchrotron yielded new spectral lines at 3141.0, 2953.6, 2911.5, 1357.4, 677.1, 895.3, and 857.0 cm(-1). These features are assigned to alpha-CH stretching (nu(1)), CH(2) antisymmetric stretching (nu(2)), CH(2) symmetric stretching (nu(3)), CH(2)-bending (nu(5)), HCCH cis bending (nu(7)), CH(2) out-of-plane bending (nu(8)), and alpha-CH out-of plane bending (nu(9)) modes of C(2)H(3), respectively, based on results of (13)C- and D-isotopic experiments and quantum-chemical calculations. These calculations using density-functional theory (B3LYP and PW91PW91/aug-cc-pVTZ) predict vibrational wavenumbers, IR intensities, and isotopic ratios of vinyl radical that agree satisfactorily with our experimental results. PMID- 18513035 TI - Photoinduced electron transfer reaction in polymer-surfactant aggregates: Photoinduced electron transfer between N,N-dimethylaniline and 7-amino coumarin dyes. AB - Photoinduced electron transfer between coumarin dyes and N,N-dimethylaniline has been investigated by using steady state and picosecond time resolved fluorescence spectroscopy in sodium dodecyl sulphate (SDS) micelles and PVP-polyvinyl pyrrolidone (SDS) polymer-surfactant aggregates. A slower rate of electron transfer is observed in PVP-SDS aggregates than in polymer-free SDS micelles. A Marcus type inversion is observed in the correlation of free energy change in comparison with the electron transfer rate. The careful investigation reveals that C-151 deviates from the normal Marcus inverted region compared to its analogs C-152 and C-481 due to slower rotational relaxation and smaller translational diffusion coefficient. PMID- 18513036 TI - Local reactivity of O2 with Pt3 on Co3Pt and related backgrounds. AB - We study the local reactivity of molecular oxygen with bimetallic substrates of a platinum trimer island supported on nanotips of CoPt, Pt, Co, Ni, and Fe. Because of the reduced interatomic distances and varying interaction strengths with the substrates, the supported island interaction with oxygen can be tuned from stronger to weaker relative to the interaction of a freestanding island with oxygen despite that there is no well-behaved trend with the binding energy of the island to the substrates. PMID- 18513037 TI - A quantum mechanical study of the reactivity of (SiO)2-defective silica surfaces. AB - The reactivity of the strained (SiO)(2)-four atom ring defect at the silica surfaces has been studied in a cluster approach adopting the ONIOM2[B3LYP6 31+G(d,p):MNDO] method to compute the ring opening reaction by interaction with H(2)O and NH(3). The vibrational "fingerprints" of the isolated defect are computed at 921, 930, and 934 cm(-1) in reasonable agreement with experimental evidence on amorphous silica outgassed at T>900 K. The opening of the (SiO)(2) four-member ring by the considered molecules is exergonic and the actual value depends on the possible constraints enforced on the reaction products by the silica surrounding. The free kinetic energy barriers result from the interplay between the nucleophilic/electrophilic character of the adsorbed molecule and are 22 and 25 kcal mol(-1) for NH(3) and H(2)O, respectively. All free energy profiles envisage an activated complex in which the nucleophilic part of the molecule interacts on the coordinatively strained silicon atom of the (SiO)(2) defect followed by the proton transfer from the coordinated molecule towards the oxygen of the defective ring. Calculations show that this step can be speed up by the presence of more than one adsorbed molecule or even more (about seven orders of magnitude), by the copresence of water molecules acting as "proton transfer helpers." In these cases, the free energy barriers decrease to approximately 13 and 15 kcal mol(-1) for NH(3) and H(2)O, respectively. For the case of H(2)O adsorption, benchmark test calculations reveal that MP2, BLYP, and B3LYP energy profiles are in very good agreement with each other, whereas for PBE, both the reaction energy and the activation barrier are underestimated. Present data also show that the molecular model mimicking the (SiO)(2) defect is far less reactive than what appears to occur on the real defect at the surface of amorphous silica. So, only a combination of some further geometrical strains imparted by the solid on the (SiO)(2) defect, not accounted for by the cluster models, and higher adsorbate loadings are needed to reharmonize experiment and simulation. Notwithstanding, the vibrational features of the reaction products have been characterized and support the available experimental measurements. PMID- 18513038 TI - Influence of the dendron chemical structure on the photophysical properties of bisfluorene-cored dendrimers. AB - A detailed study of the photophysics of a family of bisfluorene-cored dendrimers is reported. Polarized time-resolved fluorescence, singlet-singlet exciton annihilation and fluorescence quantum yield measurements were performed and used to understand how the dendron structure affects the light-emitting properties of the materials. The exciton diffusion rate is similar in all films studied. An increase in the nonradiative deactivation rate by nearly one order of magnitude is observed in films of dendrimers with stilbenyl and carbazolyl based dendrons as compared to solutions, whereas the dendrimers with biphenyl and diphenylethylenyl dendrons showed highly efficient emission (photoluminescence quantum yields of 90%) in both solution and the solid state. The results of the materials that show fluorescence quenching can be explained by the presence of quenching sites at a concentration of just a fraction of a percent of all macromolecules. A possible explanation of this quenching is hole transfer from the emissive chromophore to the dendron in a face-to-face geometry. These results are important for the design of efficient blue emitters for optoelectronic applications. PMID- 18513039 TI - Density profiles and solvation forces for a Yukawa fluid in a slit pore. AB - The effect of varying wall-particle and particle-particle interactions on the density profiles near a single wall and the solvation forces between two walls immersed in a fluid of particles is investigated by grand canonical Monte Carlo simulations. Attractive and repulsive particle-particle and particle-wall interactions are modeled by a versatile hard-core Yukawa form. These simulation results are compared to theoretical calculations using the hypernetted chain integral equation technique, as well as with fundamental measure density functional theory (DFT), where particle-particle interactions are either treated as a first order perturbation using the radial distribution function or else with a DFT based on the direct-correlation function. All three theoretical approaches reproduce the main trends fairly well, but exhibit inconsistent accuracy, particularly for attractive particle-particle interactions. We show that the wall particle and particle-particle attractions can couple together to induce a nonlinear enhancement of the adsorption and a related "repulsion through attraction" effect for the effective wall-wall forces. We also investigate the phenomenon of bridging, where an attractive wall-particle interaction induces strongly attractive solvation forces. PMID- 18513040 TI - An inherent structure view of liquid-vapor interfaces. AB - The formal statistical mechanical theory describing liquid-vapor interfaces at thermal equilibrium has been incomplete, owing partly to discrepancies between two primary views, the local free energy approach originated by van der Waals and the capillary wave approach initiated by Mandelstam. The former provides detailed prescriptions for interfacial density profiles and surface tensions, and has recently been tailored to conform to nonclassical critical phenomena. The latter postulates a crude discontinuous density profile that is geometrically delocalized by surface excitations, but which qualitatively incorporates basic gravitational effects that are missing in the van der Waals method. The present analysis provides a formalism within which both approaches can be reconciled. It draws upon the inherent structure mapping procedure to define an intrinsic liquid vapor density profile which invokes no arbitrary parameters that are not already present in the many-particle potential energy function or its thermodynamics. By construction, this intrinsic profile plays a role conventionally given to a van der Waals interface profile, and although it is free of capillary waves it can serve as a starting point for evaluating the gravitational implications of those interfacial fluctuation effects. PMID- 18513041 TI - Electrostatic Maxwell stress model of the shapes of condensed phase domains in monolayers at the air-water interface. AB - The formation mechanism of the shapes of condensed phase domains in monolayers at the air-water interface was investigated taking into account the surface pressure, line tension, and electrostatic energy due to the spontaneous polarization generated in normal and in-plane direction. By deriving the shape equation of monolayer domains as the mechanical balance at the domain boundary, we found that the electrostatic energy contributes to the shape equation as electrostatic Maxwell stress. Development of a cusp from condensed phase domains of fatty acid monolayers, which has been experimentally observed, was analyzed by the shape equation. It was found that the development of a cusp originated from the strong Maxwell stress, which was induced by the non-uniform orientational distribution in the fatty acid domain, and that cusped shapes gave a minimum of the free energy of the domain. It demonstrates that the shape equation with Maxwell stress, which is derived in the present study, is useful to study the formation mechanism of the shapes of condensed phase domains in monolayers. PMID- 18513042 TI - A kinetic theory description of the viscosity of dense fluids consisting of chain molecules. AB - An expression for the viscosity of a dense fluid is presented that includes the effect of molecular shape. The molecules of the fluid are approximated by chains of equal-sized, tangentially jointed, rigid spheres. It is assumed that the collision dynamics in such a fluid can be approximated by instantaneous collisions between two rigid spheres belonging to different chains. The approach is thus analogous to that of Enskog for a fluid consisting of rigid spheres. The description is developed in terms of two molecular parameters, the diameter sigma of the spherical segment and the chain length (number of segments) m. It is demonstrated that an analysis of viscosity data of a particular pure fluid alone cannot be used to obtain independently effective values of both sigma and m. Nevertheless, the chain lengths of n-alkanes are determined by assuming that the diameter of each rigid sphere making up the chain can be represented by the diameter of a methane molecule. The effective chain lengths of n-alkanes are found to increase linearly with the number C of carbon atoms present. The dependence can be approximated by a simple relationship m=1+(C-1)3. The same relationship was reported within the context of a statistical associating fluid theory equation of state treatment of the fluid, indicating that both the equilibrium thermodynamic properties and viscosity yield the same value for the chain lengths of n-alkanes. PMID- 18513043 TI - Shear stresses of colloidal dispersions at the glass transition in equilibrium and in flow. AB - We consider a model dense colloidal dispersion at the glass transition, and investigate the connection between equilibrium stress fluctuations, seen in linear shear moduli, and the shear stresses under strong flow conditions far from equilibrium, viz., flow curves for finite shear rates. To this purpose, thermosensitive core-shell particles consisting of a polystyrene core and a cross linked poly(N-isopropylacrylamide) shell were synthesized. Data over an extended range in shear rates and frequencies are compared to theoretical results from integrations through transients and mode coupling approaches. The connection between nonlinear rheology and glass transition is clarified. While the theoretical models semiquantitatively fit the data taken in fluid states and the predominant elastic response of glass, a yet unaccounted dissipative mechanism is identified in glassy states. PMID- 18513044 TI - A constant-time kinetic Monte Carlo algorithm for simulation of large biochemical reaction networks. AB - The time evolution of species concentrations in biochemical reaction networks is often modeled using the stochastic simulation algorithm (SSA) [Gillespie, J. Phys. Chem. 81, 2340 (1977)]. The computational cost of the original SSA scaled linearly with the number of reactions in the network. Gibson and Bruck developed a logarithmic scaling version of the SSA which uses a priority queue or binary tree for more efficient reaction selection [Gibson and Bruck, J. Phys. Chem. A 104, 1876 (2000)]. More generally, this problem is one of dynamic discrete random variate generation which finds many uses in kinetic Monte Carlo and discrete event simulation. We present here a constant-time algorithm, whose cost is independent of the number of reactions, enabled by a slightly more complex underlying data structure. While applicable to kinetic Monte Carlo simulations in general, we describe the algorithm in the context of biochemical simulations and demonstrate its competitive performance on small- and medium-size networks, as well as its superior constant-time performance on very large networks, which are becoming necessary to represent the increasing complexity of biochemical data for pathways that mediate cell function. PMID- 18513045 TI - Roles of external noise correlation in optimal intracellular calcium signaling. AB - The dynamics of a minimal calcium model, which is subjected to white noise or colored noise, was investigated. For white noise, coherence of noise-induced calcium oscillations reached a maximum at an optimal noise intensity, characterizing coherence resonance. Higher resonance peaks could be observed at lower noise intensity when a control parameter is tuned to approach a bifurcation point. For colored noise, a maximal coherence of the oscillations was found for suitable values of both the intensity and the correlation time. Moreover, the coherence of the oscillations exhibited two maxima at two values of noise intensity (correlation time) for appropriate noise correlation time (intensity). In addition, a quantitative description of the effects of noise correlation time on the resonance behavior was presented. The resonance behavior, which is induced either by white noise or colored noise, was interpreted by terms of height and relative width of a spectral peak. PMID- 18513046 TI - A Monte Carlo algorithm to study polymer translocation through nanopores. II. Scaling laws. AB - In the first paper of this series, we developed a new one-dimensional Monte Carlo approach for the study of flexible chains that are translocating through a small channel. We also presented a numerical scheme that can be used to obtain exact values for both the escape times and the escape probabilities given an initial pore-polymer configuration. We now present and discuss the fundamental scaling behaviors predicted by this Monte Carlo method. Our most important result is the fact that, in the presence of an external bias E, we observe a change in the scaling law for the translocation time tau as function of the polymer length N: In the general expression tau approximately N(beta)E, the exponent changes from beta=1 for moderately long chains to beta=1+nu or beta=2nu for very large values of N (for Rouse and Zimm dynamics, respectively). We also observe an increase in the effective diffusion coefficient due to the presence of entropic pulling on unbiased polymer chains. PMID- 18513047 TI - Inherent flexibility and protein function: The open/closed conformational transition in the N-terminal domain of calmodulin. AB - The key to understand a protein's function often lies in its conformational dynamics. We develop a coarse-grained variational model to investigate the interplay between structural transitions, conformational flexibility, and function of the N-terminal calmodulin domain (nCaM). In this model, two energy basins corresponding to the "closed" apo conformation and "open" holo conformation of nCaM are coupled by a uniform interpolation parameter. The resulting detailed transition route from our model is largely consistent with the recently proposed EFbeta-scaffold mechanism in EF-hand family proteins. We find that the N-terminal parts of the calcium binding loops shows higher flexibility than the C-terminal parts which form this EFbeta-scaffold structure. The structural transition of binding loops I and II are compared in detail. Our model predicts that binding loop II, with higher flexibility and earlier structural change than binding loop I, dominates the open/closed conformational transition in nCaM. PMID- 18513048 TI - Polymer-induced phase separation and crystallization in immunoglobulin G solutions. AB - We study the effects of the size of polymer additives and ionic strength on the phase behavior of a nonglobular protein-immunoglobulin G (IgG)-by using a simple four-site model to mimic the shape of IgG. The interaction potential between the protein molecules consists of a Derjaguin-Landau-Verwey-Overbeek-type colloidal potential and an Asakura-Oosawa depletion potential arising from the addition of polymer. Liquid-liquid equilibria and fluid-solid equilibria are calculated by using the Gibbs ensemble Monte Carlo technique and the Gibbs-Duhem integration (GDI) method, respectively. Absolute Helmholtz energy is also calculated to get an initial coexisting point as required by GDI. The results reveal a nonmonotonic dependence of the critical polymer concentration rho(PEG) (*) (i.e., the minimum polymer concentration needed to induce liquid-liquid phase separation) on the polymer-to-protein size ratio q (equivalently, the range of the polymer-induced depletion interaction potential). We have developed a simple equation for estimating the minimum amount of polymer needed to induce the liquid-liquid phase separation and show that rho(PEG) (*) approximately [q(1+q)(3)]. The results also show that the liquid-liquid phase separation is metastable for low-molecular weight polymers (q=0.2) but stable at large molecular weights (q=1.0), thereby indicating that small sizes of polymer are required for protein crystallization. The simulation results provide practical guidelines for the selection of polymer size and ionic strength for protein phase separation and crystallization. PMID- 18513049 TI - Convergence of folding free energy landscapes via application of enhanced sampling methods in a distributed computing environment. AB - We have implemented the serial replica exchange method (SREM) and simulated tempering (ST) enhanced sampling algorithms in a global distributed computing environment. Here we examine the helix-coil transition of a 21 residue alpha helical peptide in explicit solvent. For ST, we demonstrate the efficacy of a new method for determining initial weights allowing the system to perform a random walk in temperature space based on short trial simulations. These weights are updated throughout the production simulation by an adaptive weighting method. We give a detailed comparison of SREM, ST, as well as standard MD and find that SREM and ST give equivalent results in reasonable agreement with experimental data. In addition, we find that both enhanced sampling methods are much more efficient than standard MD simulations. The melting temperature of the Fs peptide with the AMBER99phi potential was calculated to be about 310 K, which is in reasonable agreement with the experimental value of 334 K. We also discuss other temperature dependent properties of the helix-coil transition. Although ST has certain advantages over SREM, both SREM and ST are shown to be powerful methods via distributed computing and will be applied extensively in future studies of complex bimolecular systems. PMID- 18513050 TI - Generalized binomial tau-leap method for biochemical kinetics incorporating both delay and intrinsic noise. AB - The delay stochastic simulation algorithm (DSSA) by Barrio et al. [Plos Comput. Biol. 2, 117(E) (2006)] was developed to simulate delayed processes in cell biology in the presence of intrinsic noise, that is, when there are small-to moderate numbers of certain key molecules present in a chemical reaction system. These delayed processes can faithfully represent complex interactions and mechanisms that imply a number of spatiotemporal processes often not explicitly modeled such as transcription and translation, basic in the modeling of cell signaling pathways. However, for systems with widely varying reaction rate constants or large numbers of molecules, the simulation time steps of both the stochastic simulation algorithm (SSA) and the DSSA can become very small causing considerable computational overheads. In order to overcome the limit of small step sizes, various tau-leap strategies have been suggested for improving computational performance of the SSA. In this paper, we present a binomial tau DSSA method that extends the tau-leap idea to the delay setting and avoids drawing insufficient numbers of reactions, a common shortcoming of existing binomial tau-leap methods that becomes evident when dealing with complex chemical interactions. The resulting inaccuracies are most evident in the delayed case, even when considering reaction products as potential reactants within the same time step in which they are produced. Moreover, we extend the framework to account for multicellular systems with different degrees of intercellular communication. We apply these ideas to two important genetic regulatory models, namely, the hes1 gene, implicated as a molecular clock, and a Her1/Her 7 model for coupled oscillating cells. PMID- 18513054 TI - Invited article: design techniques and noise properties of ultrastable cryogenically cooled sapphire-dielectric resonator oscillators. AB - We review the techniques used in the design and construction of cryogenic sapphire oscillators at the University of Western Australia over the 18 year history of the project. We describe the project from its beginnings when sapphire oscillators were first developed as low-noise transducers for gravitational wave detection. Specifically, we describe the techniques that were applied to the construction of an interrogation oscillator for the PHARAO Cs atomic clock in CNES, in Toulouse France, and to the 2006 construction of four high performance oscillators for use at NMIJ and NICT, in Japan, as well as a permanent secondary frequency standard for the laboratory at UWA. Fractional-frequency fluctuations below 6 x 10(-16) at integration times between 10 and 200 s have been repeatedly achieved. PMID- 18513055 TI - A versatile all-optical Bose-Einstein condensates apparatus. AB - We report on the construction of an all-optical Bose-Einstein condensate apparatus by using a CO2 laser trap. We also report on measurements of the trap frequency by applying a periodic perturbation to the trap potential. The derived trap parameters agree well with the design parameters. PMID- 18513056 TI - A subnanosecond pulsed ion source for micrometer focused ion beams. AB - A new, compact design of an ion source delivers nanosecond pulsed ion beams with low emittance, which can be focused to micrometer size. By using a high-power, 25 fs laser pulse focused into a gas region of 10(-6) mbar, ions at very low temperatures are produced in the small laser focal volume of 5 mum diameter by 20 mum length through multiphoton ionization. These ions are created in a cold environment, not in a hot plasma, and, since the ionization process itself does not significantly heat them, have as a result essentially room temperature. The generated ion pulse, up to several thousand ions per pulse, is extracted from the source volume with ion optical elements that have been carefully designed by simulation calculations. Externally triggered, its subnanosecond duration and even smaller time jitter allow it to be superimposed with other pulsed particle or laser beams. It therefore can be combined with any type of collision experiment where the size and the time structure of the projectile beam crucially affect the achievable experimental resolution. PMID- 18513057 TI - A broadband Fourier transform microwave spectrometer based on chirped pulse excitation. AB - Designs for a broadband chirped pulse Fourier transform microwave (CP-FTMW) spectrometer are presented. The spectrometer is capable of measuring the 7-18 GHz region of a rotational spectrum in a single data acquisition. One design uses a 4.2 Gsampless arbitrary waveform generator (AWG) to produce a 1 mus duration chirped pulse with a linear frequency sweep of 1.375 GHz. This pulse is sent through a microwave circuit to multiply the bandwidth of the pulse by a factor of 8 and upconvert it to the 7.5-18.5 GHz range. The chirped pulse is amplified by a traveling wave tube amplifier and broadcast inside the spectrometer by using a double ridge standard gain horn antenna. The broadband molecular free induction decay (FID) is received by a second horn antenna, downconverted, and digitized by a 40 Gsampless (12 GHz hardware bandwidth) digital oscilloscope. The second design uses a simplified pulse generation and FID detection scheme, employing current state-of-the-art high-speed digital electronics. In this spectrometer, a chirped pulse with 12 GHz of bandwidth is directly generated by using a 20 Gsampless AWG and upconverted in a single step with an ultrabroadband mixer. The amplified molecular emission is directly detected by using a 50 Gsampless digital oscilloscope with 18 GHz bandwidth. In both designs, fast Fourier transform of the FID produces the frequency domain rotational spectrum in the 7-18 GHz range. The performance of the CP-FTMW spectrometer is compared to a Balle-Flygare-type cavity-FTMW spectrometer. The CP-FTMW spectrometer produces an equal sensitivity spectrum with a factor of 40 reduction in measurement time and a reduction in sample consumption by a factor of 20. The CP-FTMW spectrometer also displays good intensity accuracy for both sample number density and rotational transition moment. Strategies to reduce the CP-FTMW measurement time by another factor of 90 while simultaneously reducing the sample consumption by a factor of 30 are demonstrated. PMID- 18513058 TI - Wedged multilayer Laue lens. AB - A multilayer Laue lens (MLL) is an x-ray focusing optic fabricated from a multilayer structure consisting of thousands of layers of two different materials produced by thin-film deposition. The sequence of layer thicknesses is controlled to satisfy the Fresnel zone plate law and the multilayer is sectioned to form the optic. An improved MLL geometry can be created by growing each layer with an in plane thickness gradient to form a wedge, so that every interface makes the correct angle with the incident beam for symmetric Bragg diffraction. The ultimate hard x-ray focusing performance of a wedged MLL has been predicted to be significantly better than that of a nonwedged MLL, giving subnanometer resolution with high efficiency. Here, we describe a method to deposit the multilayer structure needed for an ideal wedged MLL and report our initial deposition results to produce these structures. PMID- 18513059 TI - Optical fiber sensor for small angle and infinite angle detection. AB - An optical fiber angle sensor that consists of a cam, linear displacement attenuators, and a sensitive Fabry-Perot cavity (FPC), is demonstrated to simultaneously measure small angle and infinite angle with high resolution. When the cam is driven to rotate by a fixed rotary shaft, a linear displacement transformed from an angle will be further attenuated by linear displacement attenuators and will subsequently modulate the FPC, which causes the reflected interference intensity of FPC to vary. By using the method of interferometric fringe counting, we can measure an infinite angle accurately and continuously. Moreover, a small angle measurement can be performed in a locally linear region. Experimental results show that two resolutions of 0.5 and 1.5 s in the angle range of 0.1944 degrees and 3.5496 degrees are achieved, respectively. PMID- 18513060 TI - A synthetic heterodyne interferometer for small amplitude of vibration measurement. AB - The homodyne interferometer has the advantages of simple optical configuration and low cost. However, it requires a caution in using an electronic filter such as high pass filter (HPF) to get rid of low frequency electronic noises and the dc offset associated with the optical intensity in the photodiode electronic circuitry. When the vibration amplitude is smaller than at least 12 of the wavelength of He-Ne laser, a problem of incorrect velocity or distorted velocity measurement can be caused since a dc value of the interference signal is eliminated by using the HPF. To solve this problem of using the HPF in the homodyne interferometer, a synthetic interferometer using a mechanical modulation method is proposed in this work by exciting a reference mirror with the displacement larger than 12 of the wavelength. In this work, the analytical work is presented to show how the synthetic interferometer solves the problem of incorrect velocity measurement by using the Fourier-Bessel function description of the interference signals. Simulation and experimental works are also presented to validate the synthetic heterodyne interferometer proposed in this work. PMID- 18513061 TI - Broadband single-shot electron spectrometer for GeV-class laser-plasma-based accelerators. AB - Laser-plasma-based accelerators can provide electrons over a broad energy range and/or with large momentum spread. The electron beam energy distribution can be controlled via accurate control of laser and plasma properties, and beams with energies ranging from approximately 0.5 to 1000 MeV have been observed. Measuring these energy distributions in a single shot requires the use of a diagnostic with large momentum acceptance and, ideally, sufficient resolution to accurately measure narrow energy spread beams. Such a broadband single-shot electron magnetic spectrometer for GeV-class laser-plasma-based accelerators has been developed at Lawrence Berkeley National Laboratory. Detailed descriptions of the design concept and hardware are presented, as well as a performance evaluation of the spectrometer. The spectrometer covered electron beam energies raging from 0.01 to 1.1 GeV in a single shot, and enabled the simultaneous measurement of the laser properties at the exit of the accelerator through the use of a sufficiently large pole gap. Based on measured field maps and third-order transport analysis, a few percent-level resolution and determination of the absolute energy were achieved over the entire energy range. Laser-plasma-based accelerator experiments demonstrated the capability of the spectrometer as a diagnostic and its suitability for measuring broadband electron sources. PMID- 18513062 TI - Sub-MeV tunably polarized X-ray production with laser Thomson backscattering. AB - Reported in this article is the generation of unique polarized x-rays in the sub MeV region by means of the Thomson backscattering of the Nd:YAG laser photon with a wavelength of 1064 nm on the 150 MeV electron from the microtron accelerator. The maximum energy of the x-ray photons is estimated to be about 400 keV. The total energy of the backscattered x-ray pulse is measured with an imaging plate and a LYSO scintillator. The angular divergence of the x-rays is also measured by using the imaging plate. We confirm that the x-ray beam is polarized according to the laser polarization direction with the Compton scattering method. In addition, we demonstrate the imaging of the object shielded by lead with the generated x rays. PMID- 18513063 TI - Use of GafChromic film to diagnose laser generated proton beams. AB - A calibration of three types of GafChromic radiochromic film (HS, MD-55, and HD 810) was carried out on the Crocker Nuclear Laboratory's 76 in. cyclotron at UC Davis over doses ranging from 0.001 to 15 kGy. The film was digitized with a scanning microdensitometer with which it was scanned twice with two different filters to increase the film's effective dynamic range. We demonstrate how this calibrated film can be used to measure the spectrum and total energy of a laser generated proton beam. This technique was applied to an experiment on the 10 J, 100 fs Callisto laser at Lawrence Livermore National Laboratory. The resulting proton spectrum was compared to that obtained by simultaneous measurement of Ti nuclear activation; the two methods give the same proton beam slope temperature and agree in number of protons to within 27%. PMID- 18513064 TI - Preparation of ortho-para ratio controlled D2 gas for muon-catalyzed fusion. AB - A negative muon in hydrogen targets, e.g., D2 or D-T mixture, can catalyze nuclear fusions following a series of atomic processes involving muonic hydrogen molecular formation (muon-catalyzed fusion, muCF). The ortho-para state of D2 is a crucial parameter not only for enhancing the fusion rate but also to precisely investigate various muonic atom processes. We have developed a system for controlling and measuring the ortho-para ratio of D2 gas for muCF experiments. We successfully collected para-enriched D2 without using liquid-hydrogen coolant. Ortho-enriched D2 was also obtained by using a catalytic conversion method with a mixture of chromium oxide and alumina. The ortho-para ratio of D2 gas was measured with a compact Raman spectroscopy system. We produced large volume (5-30 l at STP), high-purity (less than ppm high-Z contaminant) D2 targets with a wide range of ortho-para ratios (ortho 20%-99%). By using the ortho-para controlled D2 in muCF experiments, we observed the dependence of muCF phenomena on the ortho para ratio. PMID- 18513065 TI - Unfolding core asymmetries with x-ray emission images in symmetry diagnostic experiments. AB - A novel inversion technique is proposed to unfold core asymmetries at the source with x-ray emission images, which were obtained from imploded surrogate capsules in symmetry diagnostic experiments. The axisymmetrical core emission can be expanded as a Fourier series, with Legendre polynomials and spherical Bessel functions as bases concerned with polar angle and radius, respectively. A least squares estimator is employed to obtain the unknown coefficients from its two dimensional image data. The unfolded Legendre coefficients can be further used to test modeling of drive asymmetries in hohlraums. This technique is also demonstrated with a proof-of-principle experiment performed on the Shenguang II laser facility [L. Zunqi et al., Chin. J. Lasers B10, 6 (2001)]. PMID- 18513066 TI - Three-electrode gas switches with electrodynamical acceleration of a discharge channel. AB - High voltage, high current, and high Coulomb transfer closing switches are required for many high power pulsed systems. There are a few alternatives for closing switches, for example, ignitrons, vacuum switches, solid-state switches, high pressure gas switches (spark gaps), and some others. The most popular closing switches up to date are spark gaps due to relatively simple design, robustness, easily field maintenance, and repair. Main drawback of spark gaps is limited lifetime, which is related directly or indirectly to erosion of the electrodes. Multichannel switches and switches with moving arc have been proposed to prevent the electrodes erosion. This study investigates switches, where a spark channel is initiated in a three-electrode layout and then the spark accelerates due to electrodynamic force and moves along the extended electrodes. At a given current amplitude, the diameter of the extended electrodes allows to control the spark velocity and hence, the erosion of the electrodes providing the required lifetime. The first switch is designed for 24 kV charging voltage and approximately 4 C total charge transfer. This spark gap was tested at 25 kA peak current in 40 000 shots in a single polarity discharge and in 20 000 shots in bipolar discharge. Second spark gap is designed for 24 kV charging voltage and approximately 70 C total charge transfer. It was tested in 22 000 shots, at a current of 250 kA with a pulse length of 360 mus. In this paper, we present design of these spark gaps and trigger generator, describe the test bed, and present the results of the tests. PMID- 18513067 TI - A compact, multiangle electron spectrometer for ultraintense laser-plasma interaction experiments. AB - Experiments on the multiterawatt (MTW) laser at the Laboratory for Laser Energetics will study the effect of the focal-spot shape on the forward acceleration and collimation of electrons. A compact electron spectrometer has been developed to record the energy spectra of electrons ejected in the interaction of the laser at multiple angular locations simultaneously. The modular system with replaceable magnets provides an adjustable energy band, currently 0.2-6 MeV. The detector is an array of imaging plates. The device is designed to operate in the high-noise environment (bremsstrahlung and Compton x rays, gamma rays, and scattered electrons), while being compact enough to fit in the 30 cm radius MTW target chamber. The detector geometry and shielding were optimized with the particle/radiation transport code GEANT4. Calibration was performed with beta sources. The required dynamic range, sensitivity, and resolution were confirmed with initial MTW experimental data. PMID- 18513068 TI - Measurement of derivative of ion temperature using high spatial resolution charge exchange spectroscopy with space modulation optics. AB - A new technique to measure the first and second derivatives of the ion temperature profile has been developed by using a charge exchange spectroscopy system with space modulation optics. The space observed is scanned up to +/-3 cm with a cosine wave modulation frequency up to 30 Hz by shifting the object lens in front of the optical fiber bundle by 0.5 mm with a piezoelement. The first and second derivatives of ion temperature are derived from the modulation component of the ion temperature measured by using Fourier series expansion. PMID- 18513069 TI - Quantitative imaging of Young's modulus of solids: a contact-mechanics study. AB - We developed equipment and methods for measuring quantitatively the local Young's modulus of solids. It consists of an electrodeless langasite oscillator and line antennas, and oscillator vibrations are generated and detected contactlessly. A constant biasing force results from oscillator mass and is independent of surface roughness. The effect of material anisotropy on the measured stiffness is theoretically discussed for studying the limitation of the quantitative measurement. The microscopy has been applied to polycrystalline copper, and the measured modulus is compared to calculations based on electron-backscatter diffraction measurements. Also, we applied it to a duplex stainless steel and an embedded silicon-carbide fiber. The results reveal textured regions, defects with high sensitivity, and even stiffness distribution in a single grain. PMID- 18513070 TI - Development of a momentum microscope for time resolved band structure imaging. AB - We demonstrate the use of a novel design of a photoelectron microscope in combination to an imaging energy filter for momentum resolved photoelectron detection. Together with a time resolved imaging detector, it is possible to combine spatial, momentum, energy, and time resolution of photoelectrons within the same instrument. The time resolution of this type of energy analyzer can be reduced to below 100 ps. The complete ARUPS pattern of a Cu(111) sample excited with He I, is imaged in parallel and energy resolved up to the photoelectron emission horizon. Excited with a mercury light source (h nu=4.9 eV), the Shockley surface state at the energy threshold is clearly imaged in k-space. Electron electron interactions are observed in momentum space as a correlation hole in two electron photoemission. With the high transmission and the time resolution of this instrument, possible new measurements are discussed: Time and polarization resolved ARUPS measurements, probing change of bandstructure due to chemical reaction, growth of films, or phase transitions, e.g., melting or martensitic transformations. PMID- 18513071 TI - Design, fabrication, and characterization of diffraction gratings for neutron phase contrast imaging. AB - We have developed a neutron phase contrast imaging method based on a grating interferometer setup. The principal constituents are two absorption gratings made of gadolinium and a phase modulating grating made of silicon. The design parameters of the setup, such as periodicity, structure heights of the gratings, and the distances between the gratings, are calculated. The fabrication of each grating is described in detail. The produced diffraction gratings were finally characterized within the setup, by locally evaluating the produced contrast (visibility) in each detector pixel, resulting in a visibility map over the whole grating size. An averaged value of 23% is achieved. PMID- 18513072 TI - Gradiometric micro-SQUID susceptometer for scanning measurements of mesoscopic samples. AB - We have fabricated and characterized micro-SQUID susceptometers for use in low temperature scanning probe microscopy systems. The design features the following: a 4.6 mum diameter pickup loop; an integrated field coil to apply a local field to the sample; an additional counterwound pickup-loop/field-coil pair to cancel the background signal from the applied field in the absence of the sample; modulation coils to allow setting the SQUID at its optimum bias point (independent of the applied field), and shielding and symmetry that minimizes coupling of magnetic fields into the leads and body of the SQUID. We use a SQUID series array preamplifier to obtain a system bandwidth of 1 MHz. The flux noise at 125 mK is approximately 0.25 mu Phi 0/ sqrt Hz above 10 kHz, with a value of 2.5 mu Phi 0/ sqrt Hz at 10 Hz. The nominal sensitivity to electron spins located at the center of the pickup loop is approximately 200 muB/ sqrt Hz above 10 kHz, in the white-noise frequency region. PMID- 18513073 TI - Construction of a low-temperature thermodynamic measurement system for single crystal of molecular compounds under pressures. AB - An apparatus to obtain low-temperature thermodynamic information under high pressures for a tiny single crystal of molecular compounds was developed based on the ac technique. To detect small temperature oscillation of a sample inside the cramp-type pressure cell, we have used a small ruthenium oxide chip sensor as a thermometer. The adoption of the four-terminal method by the ac resistance bridge has made high-resolution detection of thermal anomaly possible in the low temperature region. The constructed high-pressure thermodynamic system was mounted on a 3He refrigerator and we have succeeded to detect the thermal anomaly in relevant to magnetic order of single crystal sample of Mn4-cluster complex up to 1.05 GPa. A distinct peak of the heat capacity and its upward shift with increasing pressures was observed using a tiny crystal of about 100 microg. The high-pressure behavior of the discontinuity of heat capacity at the superconductive transition of 6 mg of metal indium has also been detected by this apparatus. The details and performance of the technique are reported. PMID- 18513074 TI - Thermodynamic measurements of submilligram bulk samples using a membrane-based "calorimeter on a chip". AB - Calorimetry offers a direct measurement of thermodynamic properties of materials, including information on the energetics of phase transitions. Many materials can only be prepared in thin film or small crystal (submilligram) form, negating the use of traditional bulk techniques. The use of micromachined, membrane-based calorimeters for submilligram bulk samples is detailed here. Numerical simulations of the heat flow for this use have been performed. These simulations describe the limits to which this calorimetric technique can be applied to the realm of small crystals (1-1000 microg). Experimental results confirm the feasibility of this application over a temperature range from 2 to 300 K. Limits on sample thermal conductivity as it relates to the application of the lumped and distributed tau 2 models are explored. For a typical sample size, the simulations yield 2.5% absolute accuracy for the heat capacity of a sample with thermal conductivity as low as 2 x 10(-5) W/cm K at 20 K, assuming a strong thermal link to the device. Silver paint is used to attach (both thermally and physically) the small samples; its heat capacity and reproducibility are discussed. Measurements taken of a submilligram single crystal of cobalt oxide (CoO) compare favorably to the results of a bulk calorimetric technique on a larger sample. PMID- 18513075 TI - A cubic boron nitride gasket for diamond-anvil experiments. AB - To maximize the thickness of the sample chamber in high-pressure experiments, we have conducted tests and have developed techniques relevant to the cubic boron nitride (c-BN) gasket for diamond-anvil cells. The c-BN gasket provides a sample chamber several times thicker than conventional metal gaskets. We have developed methods to prepare the gasket and to fill the chamber with the sample. By using the c-BN gasket, we have successfully measured x-ray diffraction patterns of SiO2 glass, a low-Z noncrystalline sample, up to 100 GPa. PMID- 18513076 TI - The compact capacitor bank CQ-1.5 employed in magnetically driven isentropic compression and high velocity flyer plate experiments. AB - Based on the low inductance capacitor, the parallel-plate transmission line, and the explosive network closing switch, a compact pulsed power generator CQ-1.5 has been developed at the Institute of Fluid Physics and is capable to deliver a current of peak of 1.5 MA within rise time of 500-570 ns into a 2-3 nH inductive load. The work is motivated to do isentropic compression experiments (ICEs) on metals up to 30-50 GPa and to launch flyer plates at velocities over 8 kms. The experiments were conducted with the diagnostics of both Doppler pin system and velocity interferometer system for any reflectors, and the measured free surface velocity histories of ICE samples were treated with a backward integration code. The results show that the isentropes of Cu and Al samples under 35 GPa are close to their Hugoniots within a deviation of 3%. The LY12 aluminum flyer plates were accelerated to a velocity over 8.96 kms. PMID- 18513077 TI - Experimental evidence of an incomplete thermalization of the energy in an x-ray microcalorimeter with a TaAu absorber. AB - We have conducted an experimental test at our XACT facility using an x-ray microcalorimeter with TaAu absorber and neutron transmutation doped germanium thermal sensor. The test was aimed at measuring the percentage of energy effectively thermalized after absorption of x-ray photons in superconducting tantalum. Moreover, in general, possible formation of long living quasiparticles implies that by using a superconducting absorber, a fraction of the deposited energy could not be thermalized on the useful time scale of the thermal sensor. To investigate this scenario, we exploited an absorber made of gold, where no energy trapping is expected, with a small piece of superconducting tantalum attached on top. We obtained evidence that the thermalization of photons absorbed in tantalum is delayed by energy trapping from quasiparticles. We compare the experimental results with numerical simulations and derive a value for the intrinsic lifetime of quasiparticles. PMID- 18513078 TI - Controlled multipulse loading with a stuffed striker in classical split Hopkinson pressure bar testing. AB - Controlled multipulse loading in classical split Hopkinson pressure bar (SHPB) testing is highly desirable for investigating loading history dependent phenomena but rarely explored. Here, we present a novel technique to achieve controlled multipulse loading in SHPB testing with a stuffed striker. This stuffed striker consists of a striker tube, and a striker bar and a gap enclosed inside the tube; upon impact on the input bar, it can produce two separated loading pulses. The gap controls the delay of the second pulse with respect to the first pulse, and the pulse separation (dwell time) can be continuously tuned from zero to hundreds of microseconds. The combination of the stuffed striker and the Lindholm technique [J. Mech. Phys. Solids 12, 317 (1964)] allows for controlled multipulse loading with triple or more pulses. We have validated the working principle of this technique with experiments, and demonstrated its feasibility and flexibility for acquiring relevant dynamic data with double- and triple-pulse loading on polycrystalline Cu. This precisely controlled multipulse loading technique is readily implementable and can be applied to investigating the dynamic response of a wide range of materials. PMID- 18513079 TI - Planar nanowire arrays formed by atomic-terrace low-angle shadowing. AB - A relatively simple method for preparation of planar nanowire arrays on vicinal substrates by molecular beam epitaxy is presented. The atomic step-and-terrace morphology of vicinal substrates is used to produce a shadowing effect on a highly collimated molecular beam at an oblique incidence to the substrate. The collimation is achieved by placing the evaporation source at a large working distance (40-100 cm) from the substrate. The method's capabilities have been demonstrated by preparation of arrays of Ag and Au nanowires on vicinal Si(111) and alpha-Al2O3 (0001) substrates. Nanowires with a width of down to 10-15 nm and a thickness of 1.5 nm have been readily achieved. PMID- 18513080 TI - Apparent volume dependence of 1/f noise in thin film structures: role of contacts. AB - The experimental investigation of low-frequency noise properties in new materials is very useful for the understanding of the involved physical transport mechanisms. In this paper it is shown that, when contact noise is present, the experimental values of the normalized Hooge parameter show a fictitious linear dependence on the volume of the analyzed samples. Experimental data on noise measurements of La0.7Sr0.3MnO3 thin films are reported to demonstrate the validity of the analysis performed. PMID- 18513081 TI - Possible application of carbon nanotube bundles for low temperature sensing. AB - We report on the R-T measurement of carbon nanotube bundles from room temperature down to 1 K. The resistance at a particular temperature depends on the diameter of the bundle. The larger the bundle diameter is, the lower the value of the resistance. The resistance increases with the decrease in temperature as in the case of carbon, carbon glass resistance thermometer, and carbon nanotubes reported in the literature. The rate of the variation of resistance depends on the resistance of the bundle at room temperature which can be explored for the low temperature thermometry. Overall, the resistance and the sensitivity of the bundle depend on the bundle diameter which can be monitored easily. PMID- 18513082 TI - A high voltage programmable ramp generator. AB - In this paper, a ramp generator with programmable slope is presented. It consists of a high voltage step generator, followed by integrator. The capacitor and inductor in the integrator are designed such that they can be varied by a microcontroller. This circuit generates two bipolar ramps with fastest speed <1 ns and provides continuous speed variation from 6 to 30 ns for a ramp of 500 V. This is being developed as a part of automated streak camera for deflection of electron beam. PMID- 18513083 TI - Stable microwave coaxial cavity plasma system at atmospheric pressure. AB - We present a systematic study of the development of a novel atmospheric microwave plasma system for material processing in the pressure range up to 760 torr and the microwave input power up to 6 kW. Atmospheric microwave plasma was reliably produced and sustained by using a cylindrical resonator with the TM(011) cavity mode. The applicator and the microwave cavity, which is a cylindrical resonator, are carefully designed and optimized with the time dependent finite element Maxwell equation solver. The azimuthal apertures are placed at the maximum magnetic field positions between the cavity and the applicator to maximize the coupling efficiency into the microwave plasma at a resonant frequency of 2.45 GHz. The system consists of a magnetron power supply, a circulator, a directional coupler, a three-stub tuner, a dummy load, a coaxial cavity, and a central cavity. Design and construction of the resonant structures and diagnostics of atmospheric plasma using optical experiments are discussed in various ranges of pressure and microwave input power for different types of gases. PMID- 18513084 TI - Microthermogravimetry using a microcantilever hot plate with integrated temperature-compensated piezoresistive strain sensors. AB - This paper reports thermogravimetric analysis of nanogram samples of paraffin using a microcantilever hot plate. The microcantilever hot plate has an integrated temperature-controlled heater and integrated temperature-compensated strain-sensing piezoresistors. The microcantilever vibration amplitude was measured using either a laser and a position sensitive photodiode, or using the piezoresistors. The cantilever resonance was measured as the cantilever was heated, such that the analyte mass could be measured as a function of temperature. Both optical and piezoresistive methods were employed to generate thermogravimetric curves for analytes in the range of 1-3 ng, and the results of the two methods compared well. PMID- 18513085 TI - Thermal conductivity measurement and interface thermal resistance estimation using SiO2 thin film. AB - In this paper, we describe an easy-to-use method to measure the thermal conductivity of thin films based on an electrical heating/sensing mechanism and a steady-state technique. The method used relative commonly used instruments, and without any signal processing circuit, is easy to be used in such thin-film thermal conductivity measurement. The SiO2 thin-film samples, prepared by thermal oxidation, plasma enhanced chemical vapor deposition (PECVD), and E-beam evaporator, were deposited on a silicon substrate. The apparent thermal conductivity, the intrinsic thermal conductivity of SiO2 films, and the total interface thermal resistance of the heater/SiO2/silicon system were evaluated. Our data showed agreement with those data obtained from previous literatures and from the 3 omega method. Furthermore, by using a sandwiched structure, the interface thermal resistance of Cr/PECVD SiO2 and PECVD SiO2/silicon were also separately evaluated in this work. The data showed that the interface thermal resistance of Cr/PECVD SiO2 (metal/dielectric) is about one order of magnitude larger than that of PECVD SiO2/silicon (dielectric/dielectric). PMID- 18513086 TI - A demountable nonmagnetic multiconductor feedthrough suitable for use in liquid helium applications. AB - A superfluid-helium-tight nonmagnetic electrical feedthrough has been developed by using brass pins embedded within an epoxy resin plug and mounted on a beryllium copper Conflat flange. A method for building these feedthroughs is discussed, and their performance history is described. PMID- 18513087 TI - Residual currents detected with a correcting electrode in a modified Bayard Alpert hot-cathode-ionization gauge. AB - A nude-type hot-cathode-ionization-gauge head with correcting electrode and shield tube, succeeded in reducing practical pressure-measurement errors caused by an influx of photoelectrons in a synchrotron radiation environment. In the development of this modified Bayard-Alpert hot-cathode-ionization gauge, experimental results showed that a basic ion current generated around the correcting electrode by emission from the gauge filament, was proportional to an emission current (less than 10% of the normal emission current) detected with a grid of correcting electrode, and that differences between estimated and measured basic ion currents in a pressure range of higher than 10(-8) Pa order meant pressure-independent residual currents detected with the correcting electrode. By using calculated forward x-ray currents, these results also showed that the estimated residual currents with the correcting electrode were mainly caused by x rays from the grid of the hot-cathode-ionization-gauge head and the grid of correcting electrode, and that the x-ray currents with the correcting electrode only caused by x rays from the grid of the hot-cathode-ionization-gauge head were about 11% of the nominal x-ray current of the ionization-gauge head. Furthermore, they may suggest that an expected residual current with the collector can be calculated by using the estimated residual current with the correcting electrode, and that this ionization gauge has an additional function to reduce pressure measurement errors caused by residual currents, though it is necessary to test the ionization gauge in an extremely high-vacuum environment under the x-ray limit of the gauge. PMID- 18513089 TI - Inductively coupled probe for the measurement of partial discharges. AB - Partial discharges are a transient phenomena whose measurement is remarkably important for electrical equipment diagnosis and maintenance. These discharges appear in the measurement circuit as very narrow current pulses of some nanoseconds. Therefore, discharge pulse detection is a particularly difficult problem, especially because they are superposed on the high voltage waveforms that cause them. In this paper, we present an inductively coupled probe able to measure this physical phenomenon by means of a very simple and inexpensive device that can be installed in the equipment under test. After modeling the probe, its response will be compared to that of commercial devices using both calibrated discharges and partial discharges occurring at real power. PMID- 18513088 TI - Ion thermalization using pressure transients in a quadrupole ion trap coupled to a vacuum matrix-assisted laser desorption ionization source and a reflectron time of-flight mass analyzer. AB - Efficient trapping and detection of intact peptide ions is demonstrated in a quadrupole ion trap (QIT) coupled to an external vacuum matrix-assisted laser desorption ionization (MALDI) source. Deactivation of metastable ions generated by MALDI is achieved in a pressure transient environment inside the QIT established by pulsing gas to access the higher pressures required for fast thermalization, without affecting vacuum conditions in the ion source and time-of flight (TOF) mass analyzer. Pressure transients are experimentally determined and a threshold of approximately 10 mTorr is identified where internally excited ions, which commonly observed to dissociate upon injection in a QIT, are stabilized. Fragment-free spectra are presented for a set of peptides by using 2,5-dihydroxybenzoic acid (DHB) as a matrix, and significantly reduced fragmentation is observed by using a-cyano-4-hydroxycinnamic acid (CHCA). Intact peptide spectra of a protein tryptic digest are also recorded with CHCA. The process of translational cooling for externally injected ions in a dynamic pressure environment is visualized by using ion trajectory simulations that employ hard sphere collisions. Statistical theory of dynamic equilibrium of ions stored in rf fields is applied to our QIT to characterize a translationally thermalized ion cloud, to explain observed ejection efficiency into the TOF mass analyzer, and to further discuss collisional deactivation of metastable ions. PMID- 18513090 TI - Very high temperature laser heated furnace for Knudsen cell mass spectrometry. AB - A very high temperature furnace (up to 3000 degrees C) for the Knudsen cell mass spectrometry (KCMS) based on a laser heating technique has been developed. It is demonstrated that this system overcomes some of the typical technological problems encountered by the standard methods and can be more easily handled in special environments such as gloveboxes or hot cells. This paper describes the laser heated KCMS general design. The technology of the laser furnace along with its advantages, disadvantages, and applications is presented. Mechanical designs, some technical details, and the importance of the temperature control are also discussed. PMID- 18513091 TI - A robust and reliable method for detecting signals of interest in multiexponential decays. AB - The concept of rejecting the null hypothesis for definitively detecting a signal was extended to relaxation spectrum space for multiexponential reconstruction. The novel test was applied to the problem of detecting the myelin signal, which is believed to have a time constant below 40 ms, in T2 decays from magnetic resonance imagining of the human brain. It was demonstrated that the test allowed the detection of a signal in a relaxation spectrum by using only the information in the data, thus avoiding any potentially unreliable prior information. The test was implemented both explicitly and implicitly for simulated T2 measurements. For the explicit implementation, the null hypothesis was that a relaxation spectrum existed that had no signal below 40 ms and that was consistent with the T2 decay. The confidence level by which the null hypothesis could be rejected gave the confidence level that there was signal below the 40 ms time constant. The explicit implementation assessed the test's performance with and without prior information where the prior information was the non-negative relaxation spectrum assumption. The test was also implemented implicitly with a data conserving multiexponential reconstruction algorithm that used left invertible matrices and that has been published previously. The implicit and explicit implementations demonstrated similar characteristics in detecting the myelin signal in both the simulated and experimental T2 decays, providing additional evidence to support the close link between the two tests. When the relaxation spectrum was assumed to be non-negative, the novel test required signal to noise ratios (SNRs) approaching 1000 in the T2 decays for detection of the myelin signal with high confidence. When the relaxation spectrum was not assumed to be non-negative, the SNR requirements for a detection with high confidence increased by a factor of 25. The application of the test to a T2 decay from human white matter, measured in vivo with a SNR of 650, demonstrated a solid detection of the signal below 40 ms believed to be due to the myelin water. This study demonstrated the robustness and reliability of extending the concept of rejecting the null hypothesis to relaxation spectrum space. The study also raised serious questions about the susceptibility to false positive detection of the myelin signal of the multiexponential reconstruction algorithms currently in use. PMID- 18513092 TI - Optimization of magneto-optical Kerr setup: analyzing experimental assemblies using Jones matrix formalism. AB - We present a comparative study on an experimental and theoretical optimization of magneto-optical Kerr setups based on photoelastic modulation and phase sensitive detector methodology. The first and second harmonics, I omega,2 omega, of the reflected light intensity are measured for a CoO/Co magnetic reference film. The magnetic field dependence of the optical off-diagonal Fresnel reflection coefficients rps and rsp follows the sample magnetization. Different Kerr setups provide various dependencies of I omega,2 omega on the reflection coefficients and, hence, on the Kerr ellipticity epsilon K and rotation theta K. Jones matrix formalism has been used to analyze the impact of a systematic variation of relative analyzer and polarizer orientations with respect to each other and with respect to the retardation axis of the modulator involved in longitudinal Kerr setups for incoming s-polarized light. We find one particular setup which maximizes I(omega) as well as I2 omega and maximizes the signal-to-noise ratio. Inefficient setups are characterized by I omega,2 omega intensities involving large nonmagnetic contributions of rp and rs. PMID- 18513093 TI - Strengthened lithium for x-ray blast windows. AB - Lithium's high x-ray transparency makes it an attractive material for windows intended to protect soft x-ray diagnostics in high energy density experiments. Pure lithium is soft and weak, but lithium mixed with lithium hydride powder becomes harder and stronger, in principle without any additional x-ray absorption. A comparison with the standard material for x-ray windows, beryllium, suggests that lithium or lithium strengthened by lithium hydride may well be an excellent option for such windows. PMID- 18513094 TI - Demonstration of a tunable two-frequency projected fringe pattern with acousto optic deflectors. AB - We report on a fringe projector for three-dimensional shape measurement. The developed instrument is able to project a two-frequency fringe pattern, each frequency is independently controlled by electronics. Moreover, each phase of the two fringe patterns is also independently adjusted. The projection system is based on the use of a pair of custom large bandwidth (40 MHz) and high efficiency (60%) TeO2 deflectors. The developed instrument offers the combined advantages of a static two-frequency fringe projector and of a tunable single frequency fringe projector. PMID- 18513095 TI - Ultrahard diamond indenter prepared from nanopolycrystalline diamond. AB - Knoop indenters were prepared from nanopolycrystalline diamonds (NPDs) synthesized by direct conversion sintering from graphite under high pressure and high temperature. Owing to the fine structure (grain size: 10-100 nm) of NPD, high-accuracy sharp edges could be formed at the indenter tips. The indentation tests demonstrated that the NPD indenter can form normal (measurable) indentations on NPD samples without fracture or chipping even at high temperatures of up to 1000 degrees C, while conventional indenters made of single crystal diamonds break easily above 600 degrees C. This suggests that the NPD indenter has greater potential in high-temperature hardness tests than the conventional single-crystal diamond indenters. PMID- 18513096 TI - Setup for in situ deep level transient spectroscopy of semiconductors during swift heavy ion irradiation. AB - A computerized system for in situ deep level characterization during irradiation in semiconductors has been set up and tested in the beam line for materials science studies of the 15 MV Pelletron accelerator at the Inter-University Accelerator Centre, New Delhi. This is a new facility for in situ irradiation induced deep level studies, available in the beam line of an accelerator laboratory. It is based on the well-known deep level transient spectroscopy (DLTS) technique. High versatility for data manipulation is achieved through multifunction data acquisition card and LABVIEW. In situ DLTS studies of deep levels produced by impact of 100 MeV Si ions on Aun-Si(100) Schottky barrier diode are presented to illustrate performance of the automated DLTS facility in the beam line. PMID- 18513097 TI - Analytical evaluation of describing functions arising from harmonic balance analysis of tapping mode atomic force microscope. AB - A new algorithm of harmonic balance analysis of tapping mode atomic force microscopes has been developed. The new algorithm is applicable to analytical evaluation of a large class of common tip-sample interaction potentials. The extensive test calculations show that the proposed algorithm in this work is the efficient one in practical computations. The comparative values presented in tables are acceptable and have the excellent agreement with the numerical results. PMID- 18513098 TI - Mass measuring instrument for use under microgravity conditions. AB - A prototype instrument for measuring astronaut body mass under microgravity conditions has been developed and its performance was evaluated by parabolic flight tests. The instrument, which is the space scale, is applied as follows. Connect the subject astronaut to the space scale with a rubber cord. Use a force transducer to measure the force acting on the subject and an optical interferometer to measure the velocity of the subject. The subject's mass is calculated as the impulse divided by the velocity change, i.e., M=integral Fdt/delta v. Parabolic flight by using a jet aircraft produces a zero-gravity condition lasting approximately 20 s. The performance of the prototype space scale was evaluated during such a flight by measuring the mass of a sample object. PMID- 18513099 TI - Measurement of edge density profiles of Large Helical Device plasmas using an ultrashort-pulse reflectometer. AB - We report here on the application of an ultrashort-pulse reflectometer (USPR) to Large Helical Device in National Institute for Fusion Science. An impulse with picosecond pulse width is used as a source in an USPR. Since the bandwidth of a source is inversely related to the pulse width, we can utilize the frequency range of microwave to millimeter-wave by using wide band transmission lines. The density profiles can be reconstructed by collecting time-of-flight signal of each frequency component of an impulse reflected from each cutoff layer. Remote control system using super science information network has been introduced to the present USPR system. PMID- 18513100 TI - GaInSn usage in the research laboratory. AB - GaInSn, a eutectic alloy, has been successfully used in the Magneto-Thermofluid Research Laboratory at the University of California-Los Angeles and at the Princeton Plasma Physics Laboratory for the past six years. This paper describes the handling and safety of GaInSn based on the experience gained in these institutions, augmented by observations from other researchers in the liquid metal experimental community. GaInSn is an alloy with benign properties and shows considerable potential in liquid metal experimental research and cooling applications. PMID- 18513102 TI - New methods for practice oriented post-graduate medical education: training of a ski-camp doctor in Hungary. PMID- 18513103 TI - Introduction to: new methods for practice oriented post-graduate medical education: training of a ski-camp doctor in Hungary. PMID- 18513104 TI - Delayed acclimatization of the ventilatory threshold in healthy trekkers. AB - OBJECTIVE: To test the hypothesis that acclimatization to high altitude results in an improvement of the ventilatory threshold (VT). METHODS: Eight lowlanders underwent cardiopulmonary exercise testing with a cycle ergometer to determine VT and peak oxygen uptake (Vo2peak) in Coventry, United Kingdom (altitude: 80 m), on arrival in leh, india (altitude: 3500 m), and after 12 days of acclimatization that included a 5-day high altitude trek up to 4770 m. RESULTS: Vo2peak fell on arrival at 3500 m and remained depressed at 12 days. VT was depressed on arrival at high altitude and was further depressed at 12 days. VT as a proportion of the Vo2peak was decreased on arrival at high altitude, and after acclimatization, this relationship was further decreased. CONCLUSIONS: Individuals who are sedentary or not participating in regular physical training appear to require a longer period of acclimatization than trained athletes. With the increasing numbers participating in high-altitude trekking and charity climbs of peaks, such as Mt. Kilimanjaro, this information has clinically significant practical implications for those leading or acting as medical advisors. PMID- 18513105 TI - Injury and illness at the Newport-Bermuda race 1998- 2006. AB - OBJECTIVE: To describe the relative frequency and types of injury and illness in the Newport-Bermuda off-shore yachting race. METHODS: At the end of each race held in even numbered years from 1998-2006, the captain of each boat was asked to complete a survey detailing any injury or illness among his/her crew. RESULTS: There was an overall 87% response rate to the survey. During the study period, 38 injuries and 57 illnesses were reported for an estimated 8105 sailors, yielding rate of injury or illness of 12 per 1000 races per sailor. Most common were injuries to the upper extremity (47%), and lacerations were the most common type of injury (45%). Sea sickness was the most common illness, and the rate of illness and injury increased in races that took place in heavy weather. Radio consultations were used 4 times, and 3 sailors required transport to a hospital. CONCLUSIONS: The rate of injury and illness was relatively low in the Newport Bermuda race. Injuries to the upper extremities and lacerations were most common, and sea sickness was the most common illness. The majority of illness and injury can be initially managed onboard. PMID- 18513106 TI - The use of extrication devices in crevasse accidents: official statement of the International Commission for Mountain Emergency Medicine and the Terrestrial Rescue Commission of the International Commission for Alpine Rescue intended for physicians, paramedics, and mountain rescuers. AB - Injured patients in crevasses who are suspected of having sustained spinal injuries should ideally be extricated after being immobilized in a horizontal position on a stretcher and having a cervical collar applied. Sometimes, however, horizontal stabilization is not possible, because the crevasse is too narrow, and the patient needs to be stabilized in a vertical position. In such cases an extrication device can be a useful adjunct. The Kendrick Extrication Device stabilizes the position of the body and maintains firm support of the head, neck, and torso. Therefore, the International Commission for Mountain Emergency Medicine supports the use of this device in narrow crevasses, if horizontal evacuation is not possible. PMID- 18513107 TI - The "worldwide shortage" of antisnake venom: is the only right answer "produce more" or is it also "use it smarter?". AB - A frequent tenet of snakebite literature is what has been described as the "worldwide shortage of antisnake venom" (ASV) and the demand for greater production. Antisnake venom is the mainstay of snakebite management, and thus this principle of "shortage" can impact the view of policy makers when it comes to framing solutions to the problem. This paper presents a model to enable policy makers to assess the amount and utilization of ASV in their areas. The model assesses ASV usage according to 2 criteria: risk and wastage. The actual usage of ASV is segmented in the model into the following main areas: (1) victims who receive too little ASV (high risk/low wastage); (2) victims who receive ASV either unnecessarily or in too great a quantity (low risk/high wastage); (3) victims who receive ASV that is not effective (high risk/high wastage); (4) victims who receive ASV according to effective local protocols (low risk/low wastage). The current proposition about the "shortage" of ASV and the proposal to simply produce more addresses only a small part of the high-risk/low-wastage group and does not address the 2 high-wastage groups. Until the high-wastage groups are recognized and resolved with training and local protocols and moved into the low-risk/low-wastage group, the true requirement for ASV worldwide cannot be assessed. PMID- 18513108 TI - Tegu (teiu) bite: report of human injury caused by a Teiidae lizard. AB - Lizards of the Teiidae family are large reptiles measuring up to 2 meters long. If threatened, they can demonstrate aggressive behavior by whipping their tail and occasionally biting. Here, we report a severe injury following a Teiidae lizard bite on the right index finger of a human. There was significant soft tissue damage and an avulsion fracture of the distal phalanx. He was treated with conservative wound care and prophylactic antibiotics. He developed no evidence of secondary infection and underwent delayed skin grafting. PMID- 18513109 TI - Marine catfish sting causing fatal heart perforation in a fisherman. AB - Many marine catfish have serrated bony stings ("spines"), which are used in defense against predators, on the dorsal and pectoral fins. While catfish-induced injuries are generally characterized by the pain associated with envenomation, the stings in some species are sufficiently long and sharp to cause severe penetrating trauma. Most injuries are to the hands of victims, commonly fishermen. We report the death of a fisherman caused by myocardial perforation from a catfish sting. To our knowledge, this is the first such description in the medical literature. PMID- 18513110 TI - Possible anaphylactic reaction due to pulmonary hydatid cyst rupture following blunt chest trauma: a case report and review of the literature. AB - A hydatid cyst is a parasitic disease caused by the tapeworm Echinococcus granulosus. It is endemic in many areas, including New Zealand, Australia, and the Mediterranean region. Pulmonary hydatid disease can be diagnosed incidentally in asymptomatic patients or may cause symptoms such as cough, chest pain, dyspnea, fever, and hemoptysis both in patients with ruptured and nonruptured cysts. Anaphylactic reaction is a rare presentation of pulmonary hydatid cyst disease. In this case report, we report an unusual anaphylactic reaction following pulmonary hydatid cyst rupture secondary to blunt chest trauma. PMID- 18513111 TI - Intravenous fluid warming with body contact in a wilderness setting. PMID- 18513113 TI - Rope tangling injuries--how should a climber fall? PMID- 18513114 TI - The effect of ambient temperature on the use of supplemental oxygen at high altitude. PMID- 18513115 TI - Efficacy of a new blister prevention plaster under tropical conditions. AB - OBJECTIVE: To survey the incidence of blister formation in a group of military recruits and to determine the efficacy of a new commercially available blister prevention plaster (Blist-O-Ban). METHODS: Questionnaires were completed by 100 male recruits, and the same 100 were entered into a field trial. Due to illness, 2 did not complete the study. Premarch foot condition was documented. Each participant acted as his own control. Foot side and plaster site were randomly selected, and plasters were applied according to a strict protocol. Participants were re-examined for blisters after a 16-km hike with pack, and the condition of the feet was documented. A chi2 test was used to analyze the results. RESULTS: Ninety-eight volunteers with 101 Blist-O-Ban application sites participated. Twenty-seven participants developed a total of 46 blisters. Heel and toe blisters accounted for more than half of all blisters. None of the bandage application sites developed blisters, and 99 out of 101 bandages remained well adhered to the application sites. CONCLUSIONS: When applied according to strict protocol, the new blister prevention bandage remained well-adhered to application sites and prevented the formation of foot blisters even in humid tropical conditions. PMID- 18513116 TI - Risk factors for coliform bacteria in backcountry lakes and streams in the Sierra Nevada mountains: a 5-year study. AB - OBJECTIVE: To provide a 5-year longitudinal assessment of risk of acquiring disease from Sierra Nevada Wilderness area lakes and streams. This study examines the relative risk factors for harmful water microorganisms, using coliforms as an indicator. METHODS: Streams and lakes in the backcountry of Yosemite and Kings Canyon National Parks and neighboring wilderness areas were selected and water was analyzed each year over a 5-year period. A total of 364 samples from lakes or streams were chosen to statistically differentiate the risk categories based on land usage, as follows: 1) areas rarely visited by humans (Wild), 2) human day use-only areas (Day Hike), 3) areas used by backpackers with overnight camping allowed (Backpack), 4) areas primarily impacted by horses or pack animals (Pack Animal), and 5) cattle and sheep grazing tracts (Cattle). Water was collected in sterile test tubes and Millipore coliform samplers. Water was analyzed at the university microbiology lab, where bacteria were harvested and then subjected to analysis using standardized techniques. Statistical analysis to compare site categories was performed utilizing Fisher exact test and analysis of variance. RESULTS: A total of 364 sampling sites were analyzed. Coliforms were found in 9% (4/47) of Wild site samples, 12% (5/42) of Day Hike site samples, and 18% (20/111) of Backpacker site samples. In contrast, 63% (70/111) of Pack Animal site samples yielded coliforms, and 96% (51/53) of samples from the Cattle areas grew coliforms. Differences between Backpacker vs Cattle or Pack Animal areas were significant at P